Preparation of silk fibroin carriers for controlled release

Preparation of Silk Fibroin Carriers for Controlled Release QIANG LIU,1HAIFENG LIU,1*AND YUBO FAN1,2*

1Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education,School of Biological Science and Medical Engineering,Beihang University,Beijing100191,People’s Republic of China

2National Research Center for Rehabilitation Technical Aids,Beijing100176,People’s Republic of China KEY WORDS silk?broin;carrier;controlled release

ABSTRACT Silk?broin provides a new option for controlled release systems as a result of its excellent biodegradability,biocompatibility,and mechanical properties.As the core material, silk?broin can be designed and widely used in drug/gene delivery,regenerative medicine,and other biomedical?elds.This review focuses on the preparation methods,loading molecules,and applications of silk?broin-based controlled release systems including microspheres,microcap-sules,?lms,microparticles,microneedles,liposomes,and hydrogels.These systems provide numerous advantages such as high encapsulation ef?ciency,avoiding loss of bioactivity and maintaining desirable range without peaks and valleys in comparison to the traditional adminis-tration approaches.Microsc.Res.Tech.00:000–000,2015.V C2015Wiley Periodicals,Inc.

INTRODUCTION

Many biomedical applications need for the delivery of bioactive molecules and drugs in slow,sustained, controlled release formats.Controlled release systems provide a variety of advantages over traditional peri-odic systemic administration,which contain increased ef?cacy,reduced or eliminated unwanted side effects, improved tissue regeneration and bioavailability,and maintained the molecule levels in a controllable range without peaks and valleys(Liu et al.,2007a;Numata et al.,2010;Pritchard et al.,2011).Moreover,such delivery systems will be bene?cial if they are biode-gradable,biocompatible,and mechanically durable and can be prepared and processed under ambient aqueous conditions to avoid loss of bioactivity of the delivered bioactive molecules.

Silk?broin is a biologically derived protein polymer isolated from domestic silkworm cocoons,which has been investigated as a biomaterial for tissue engineering and controlled release systems(Liu et al.,2007b,a;Yucel et al.,2009).Silk?broin represents a unique family of natural?brous proteins owing to their unique structure and resulting https://www.sodocs.net/doc/309243566.html,pared to other natural degradable materials,silk?broin exhibits excellent bio-compatibility with low in?ammatory and immunogenic response,robust mechanical properties,and tunable deg-radation rates ranging from weeks to years(Liu et al., 2008b,a,b;Gong et al.,2014).Based on these characteris-tics,silk?broin has been developed into many formats such as microspheres,microcapsules,microparticles, microneedles,liposomes and hydrogels,which can be loaded with drugs,cells,growth factors,and other biomo-lecules.The silk?broin carriers exhibit high encapsula-tion ef?ciency and controllable release kinetics as a result of the control of crystalline b-sheet formation (Wang et al.,2010),which gives rise to a controlled release in vitro and in vivo.Moreover,these carries based on silk?broin can ameliorate numerous problems such as a rapid breakdown of plasma concentration, potential tissue damages from extravasation,and the lack of selectivity for target cells(Pritchard et al.,2011). Thus,it is very important to?nd a correlation by model-ing or other way between dissolution rate of drug/cell in vitro and metabolism dynamics in vivo.

Controlled release carriers made of silk?broin can be prepared by numerous techniques.Their choice relies on the mode of applications,desired release kinetics,and process ability.Preparation methods include exposure of silk?broin to heat,pH extremes, organic solvents,drying,and freezing.When sensitive biologicals such as therapeutic proteins,peptides,and growth factors are incorporated,high temperature and organic solvents must be avoided for the purpose of maintaining their biological potency(Wenk et al., 2011).Various methods used for the preparation of silk ?broin controlled release systems will be discussed in detail below.

CHARACTERISTICS OF SILK FIBROIN FOR

CONTROLLED RELEASE

Effective polymer-based delivery systems need to be biodegradable and biocompatible,while they should have suitable mechanical properties,demand mild processing conditions,and afford controlled release.

*Correspondence to:Haifeng Liu and Yubo Fan;School of Biological Science and Medical Engineering,Beihang University,Xue Yuan Road No.37,Haidian District,Beijing100191,People’s Republic of China.E-mail:haifengliu@buaa. https://www.sodocs.net/doc/309243566.html, and yubofan@https://www.sodocs.net/doc/309243566.html,

Received31August2015;accepted in revised form9November2015 REVIEW EDITOR:Dr.Mingying Yang

Contract grant sponsor:National Key Technology R&D Program;Contract grant numbers:2012BAI18B06,2014BAI11B02,2014BAI11B03;Contract grant sponsor:National Natural Science Foundation of China;Contract grant num-bers:31470938,81171473,11421202,61227902,and11120101001;Contract grant sponsor:International Joint Research Center of Aerospace Biotechnol-ogy and Medical Engineering from Ministry of Science and Technology of China;Contract grant sponsor:111Project;Contract grant number: B13003;Contract grant sponsor:Doctoral Program of Higher Education of China;Contract grant number:20131102130004;Contract grant sponsor: Central Universities in Beijing;Contract grant number:ZDZH20141000601. DOI10.1002/jemt.22606

Published online00Month2015in Wiley Online Library(https://www.sodocs.net/doc/309243566.html,).

V C2015WILEY PERIODICALS,INC.

MICROSCOPY RESEARCH AND TECHNIQUE00:00–00(2015)

Silk?broin is a naturally derived protein polymer that can meet these requirements(Numata et al.,2010).

Molecular Properties

The silk?broin?bers from Bombyx mori consist of three structural proteins,including the?broin heavy chain(2390kDa),light chain(226kDa),and a small glycoprotein named P25(of ca30kDa).The heavy chain of silk?broin has an amphiphilic nature,containing hydrophobic and hydrophilic blocks(Kundu et al.,2013; Omenetto et al.,2010).The hydrophobic blocks contain a repeating sequence of Gly–Ala–Gly–Ala–Gly–Ser, which play an important role on generating the crystal-line structure of silk?broin via folding into b-sheet. However,compared with the hydrophobic region,the hydrophilic region is a short and nonrepetitive segment (Bini et al.,2004;Kundu et al.,2014).Therefore,silk ?broin cannot be dissolved in water,but it can be modi-?ed by potential reactive side groups,such as alcohols, phenols,amines,and carboxyl groups.Such modi?ca-tions can give rise to change in silk?broin hydrophilic-ity and porosity.A variety of drugs/genes can be loaded and controlled by different functional groups and dis-tinct degree.

Crystallinity

Silk?broin is a semicrystalline biopolymer which is made up of crystalline and amorphous regions.The crystalline part has two prominent structures includ-ing silk I and silk II(Hofmann et al.,2006),also known as a and b type.Silk I derived from spinning dope is unstable and water-soluble,but silk I can be fabricated more stable when it is transformed into silk II by the spinning process.The crystalline structure of B.mori ?bers is made up of silk II structure due to its b-sheet conformation,while the amorphous part of silk?broin is in random coil conformation.It is reported that the b-sheet contains an asymmetrical structure featuring hydrogen side chains(Hofmann et al.,2006;Asakura et al.,2001),consisting of glycine and alanine.The chains are held together via strong interactions with hydrogen bonds between the methyl and hydrogen groups(Bourne et al.,2003).The strong hydrogen bonds and van der Waals forces are generated,which gives rise to stack in the crystals and obtain a stable structure.Thus,the crystalline structure of silk?broin can produce physical and chemical properties such as high strength and resistance to chemicals and low elasticity and extensibility.For instance,Jin et al. (2005)reported that a treatment with water vapor made silk?broin maintain better elasticity than a treatment with methanol.However,Wang et al.(2007) reported that the microparticles treated with methanol were more stable in comparison with the micropar-ticles treated with NaCl.Thus,the properties with crystallinity are bene?cial for controlled release via using different condition.

Mechanical Properties

Some previous studies fabricated scaffolds with colla-gen and PLGA for controlled release,but these scaffolds lacked abundant mechanical strength for the purpose of loading with several drug molecules(Holland et al., 2006).Therefore,enhancing its mechanical strength is extremely necessary when using drug delivery device as a scaffold with load-bearing function.For example, as needed in bone repair,the physical properties of col-lagen are often cross-linked to enhance mechanical strength.Nevertheless,several adverse effects such as cellular toxicity and in?ammatory responses on adja-cent native tissue may be produced owing to the use of cross-linking agent.Silk?broin from B.mori shows out-standing mechanical properties,which can avoid being cross-linked due to robust b-sheet conformation(Siri-chaisit et al.,2003;Shao et al.,2002;Du et al.,2011).In addition,it was reported that water-stable aqueous-derived silk?broin scaffolds exhibited superior mechan-ical properties in comparison with the mechanical prop-erties of some natural polymeric scaffolds.The compressive strength of the silk?broin scaffolds was increased with the increasing of silk?broin concentra-tion,the uniform distribution of pores,and the decreas-ing of pore size(Bhrany et al.,2008;Wenk et al.,2011).

Biodegradation

To develop a successful formulation of controlled release,it is necessary to take into account its biode-gradation rate.The biodegradation rate is affected by numerous factors,which include diffusion through the polymer wall,desorption of surface bond,and com-bined erosion(Soppimath et al.,2001).Generally,the sensitivity of synthetic polymers to enzymatic degra-dation is lower than natural polymers such as?brino-gen,collagen,and hyaluronic acid(Dawson et al., 2008).As a natural polymer,silk?broin usually goes through proteolytic degradation,which results in non-toxic by-products.It was reported that the hydrolysis degradation was induced by immersing freeze-dried silk?broin scaffolds in phosphate-buffered saline solu-tions(Mottaghitalab et al.,2015).In addition,Wang et al.(2008)demonstrated that the silk?broin porous scaffolds were not only biodegradable but also biore-sorbable as a result of degradation by macrophages.In a word,we should take into consideration the possibil-ity of enzymatic,surface-mediated biodegradation, and controlled biodegradation of silk?broin as a deliv-ery carrier.

PREPARATION OF SILK FIBROIN CARRIERS FOR CONTROLLED RELEASE

Novel devices and technologies are demanded for controlled release of molecules so as to reduce their side effects,optimize their ef?cacy,and enhance patient compliance.Currently,the employ of nanotech-nology has given rise to the development of numerous new carriers capable of controlled release(Hosseinza-deh et al.,2012;Tahamtan et al.,2014).These devices have numerous forms,which include microspheres, microcapsules,hydrogel,liposomes,microneedles,and ?lms.Among them,the carries based on silk?broin have a wide range of applications in drug/gene/tar-geted delivery,tissue engineering,and regenerative medicine(Meinel et al.,2012).

Silk Fibroin Microspheres Microspheres are de?ned as the bioactive molecules dissolved or dispersed in the polymer matrix to form spherical particles,which possess the sustained release

2Q.LIU ET AL.

Microscopy Research and Technique

effect and are capable of targeting to speci?c tissues or organs.Silk?broin microspheres can be fabricated using numerous methods such as emulsi?cation diffu-sion(Cheerarot et al.,2015),silk/PVA blend?lms (Wang et al.,2010),spray-drying(Tomoaki et al.,2003), and micro?uidics(Breslauer et al.,2010).Currently, silk?broin microspheres can be devised as some new controlled drug/gene delivery systems.The ability to control the release pro?les by means of the processing strategies can remain the bioactivity of loaded drugs or genes.

Emulsi?cation Diffusion.Silk?broin micro-spheres can be prepared by traditional method with water-in-oil emulsion solvent diffusion(Kakran et al., 2014).Srisuwan et al.fabricated the silk?broin micro-spheres based on water-in-oil emulsi?cation diffusion with liquid paraf?n as the continuous phase.This pro-cess was characterized by encapsulation ef?ciency and controllable release kinetics,which was tunable through silk concentration.The prepared silk?broin microspheres were spherical shape in the size ranges of80–150l m and the conformation was transformed from coil to b-sheet with the methanol treatment. They could adsorb bovine serum albumin and their ef?ciency of adsorption was improved with reducing the silk?broin microsphere size(Srisuwan et al., 2009).In another study,silk?broin microspheres were loaded with3T3cells,and the effects of cellular uptake of silk?broin microspheres on the growth and cell cycle progression of3T3cells were investigated by Nam et al.(2015).The results indicated that the cellu-lar uptake of silk?broin did not affect cellular viabil-ity,which could be a potential vehicle for intracytoplasmic delivery.

The Blend Membrane.Microspheres can be fab-ricated by blend membrane based on phase separation. The blend membrane refers to mixing the protein and organic solvent and then drying the mixture into a ?lm,but the?lm should be dissolved in water for the purpose of removing organic solvent.Wang et al.suc-cessfully prepared the microspheres using the blend membrane method with silk?broin and polyvinyl alco-hol.The size of the microspheres was in the range of 300nm up to20l m.The results showed that the microsphere size and polydispersity could be controlled either by changing the concentration of silk?broin or by using ultrasonication.The silk?broin microspheres could be loaded with tetramethyl rhodamine conju-gated bovine serum albumin,dextran,and rhodamine B,and the loading ef?ciency was about95%(Wang et al.,2010).In another study,Chen et al.prepared the recombinant bone morphogenetic protein-2/silk?broin (rhBMP-2/SF)microspheres to enhance the bioactivity of biphasic calcium phosphate(BCP).The ef?cacy of BCP with these rhBMP-2/SF microspheres were eval-uated in a sheep lumbar fusion model.The in vivo results showed that the silk?broin microspheres including a very low dose of rhBMP-2could improve fusion in sheep using BCP constructs(Chen et al., 2015).

Self-Assembly.Self-assembly is a process in which a disordered system of pre-existing ingredients forms an organized structure or pattern due to speci?c,local interactions among the ingredients themselves.Many silk?broin microspheres can be prepared using self-assembly method(Rammensee et al.,2008;Carl et al., 2014).Cao et al.developed an easy and ef?cient method using self-assembling to fabricate regenerated silk?broin microspheres.The results showed that the regenerated silk?broin microspheres with sizes rang-ing from0.2to1.5l m could be in?uenced by the freez-ing temperature and ethanol concentration.When the ratio of regenerated silk?broin was increased from 2:20to8:20,the size and the polydispersity index of microspheres became smaller and narrower.While the regenerated silk?broin mixture solution was frozen, the gel particle was retarded further aggregation(Cao et al.,2007).This method did not use any cross-linking agent,toxic organic reagent,or surfactant that would adversely in?uence the living body.

Spray-Drying.Microspheres can also be fabri-cated by spray-drying method,which is emerged as materials are dispersed into particles like fog and transformed into powder when amount of water is removed by hot air.The microspheres prepared by spray-drying with mild conditions possess high encap-sulation ef?ciency and narrow size distribution.But the yield of microsphere is decreased as the particles adhere to the inner wall of dryer(Zhu et al.,2015). Tomoaki et al.exposed the silk?broin microspheres obtained by spray-drying method to atmosphere of 89%RH.The results showed that the silk micro-spheres were sphere shaped with mean diameter of5 l m and the secondary structure of silk?broin changed from b-sheet to irregular structure.The silk?broin microspheres could be loaded with theophylline and were found to be amorphous in the microsphere (Tomoaki et al.,2003).In another study,Farrago et al. used spray-drying method to prepare silk?broin microspheres by adding calcium gluconate as a cal-cium source for alginate cross-linking.The results indicated that the silk?broin microspheres with cross-linking agent could be potentially applied for delivery systems and the treatment of pressure ulcers(Farago et al.,2015).

Micro?uidics.Micro?uidic devices are based on the operation of continuous liquid?ow via microfabri-cated channels,which are conducted by external pres-sure sources,integrated mechanical micropumps,or capillary forces.Silk?broin microspheres fabricated by micro?uidic devices had a nearly homogeneous size distribution and the size could be controlled by the ?ow rate ratio(Rammensee et al.,2008;Alexander et al.,2014).Breslauer et al.successfully fabricated silk?broin microspheres using the micro?uidic method.However,these silk?broin microspheres could give rise to rapid coagulation when the methanol content in the continuous phase was increased above 25wt%.Therefore,there was a competition that should be carefully balanced between the advection of silk solution droplets down the device,the coagulation, and the crystallization(Breslauer et al.,2010).

Till now,silk?broin microspheres have been proc-essed using a variety of common approaches,including water-in-oil emulsi?cation,self-assembly,spray-dry-ing,blend membrane,and several methods rarely used such as acetone mix/dialysis,capillary-microdot,and solution mixed with combining the cylindrical steel mold.These methods are exhibited in Table1.The size of silk?broin microspheres is hard to be controlled

PREPARATION OF SILK FIBROIN CARRIERS3 Microscopy Research and Technique

owing to the nonuniform size of droplets.However,the fabrication method using micro?uidic chip provides the future perspectives due to a nearly homogeneous size distribution.

Silk Fibroin Microcapsules Microcapsule has a typical core–shell structure, which is a small sphere with a uniform wall around it. Microcapsules can be divided into two chief classi?ca-tions,including mononuclear and multinuclear micro-capsules.Mononuclear microcapsules consist of one spot core droplets,while multinuclear microcapsules are made up of multiple small entities core droplets involved in a single polymer shell.Because of their core–shell structure,microcapsules can be loaded with biomolecules,deliver them,and preserve their potency (Wenk et al.,2011).

Materials for Microcapsulations.It is the pro-cess by which individual particles or droplets of solid or liquid material are surrounded or coated with a con-tinuous?lm of polymeric material to prepare capsules in the micrometer-to-millimeter range,known as microcapsules.Microcapsules are fabricated with a variety of materials,including semisynthetic poly-mers,synthetic polymers,and natural polymers, which were shown in Table2.

Complex Coacervation.Microcapsules with the size of1–100l m are made up of a distinct capsule wall,encompassing an active core(Tharwat et al., 2013).Deveci et al.successfully fabricated the micro-capsules by complex coacervation of silk?broin and chitosan.n-Eicosane was employed as the core mate-rial and the encapsulation of core material was deter-mined via energy dispersive spectrometer(EDS) analysis.The results showed that microcapsules had smooth outer surface when the ratio of silk?broin to chitosan was close to5and they could effectively encapsulate n-Eicosane.This study con?rmed that the silk?broin/chitosan wall system could be employed to encapsulate a phase change material(Deveci et al., 2009).In addition,Nayak et al.(2014)prepared silk ?broin microcapsules to load with live cells for poten-tial therapeutic application,especially for hepatocytes transplantation in acute liver failure.

Layer-by-Layer Assembly.A series of approaches are applied for silk?broin microcapsules preparation, such as traditional layer-by-layer(LBL)assembly,self-assembly,and oil-in-water emulsion.However,there are numerous issues connected with these preparation methods such as aggregation,nonuniform size,and high polydispersity.At present,the approach with novel LBL assembly has been broadly utilized as drug/ gene delivery vehicles owing to a simple and high yield route(Shen et al.,2015).Shchepelina et al.developed a method to prepare silk?broin microcapsules with good stability,high permeability,and monodisperse in size.The silk?broin microcapsules prepared by LBL assembly composed of5,8,10,and12silk?broin layers and could be controlled by the thickness of the shell(Shchepelina et al.,2011).In another study,silk ?broin LBL microcapsules loaded with plasmid DNA were prepared by pre-and post-loading techniques. Plasmid DNA loading ef?ciency depended on the mode of plasmid DNA loading,their release rate was in?u-enced by the presence of proteolytic enzymes in release medium.The results indicated that silk?broin

TABLE1.Silk?broin-based microspheres for controlled release

Preparation Dimension Loaded molecule Key?ndings Reference

Bead milling200nm Bead Narrow size distribution of SF

particles at pH10Kazemimostaghim et al.,2013

Spray-drying5l m Theophylline Change in the secondary

structure of silk?broin

Tomoaki et al.,2003

Solution mixed with

combining the cylindrical steel mold 5–18l m Sema3A Supporting the growth of MC3T3

cells

Wang et al.,2015

Capillary-microdot<100nm Curcumin Curcumin loaded beyond8

weeks

Numata et al.,2009

Micro?uidics1–10l m-Obtaining a nearly homogeneous

size distribution

Breslauer et al.,2010

Self-assembly-RhBMP-2Sustained-release for more than

28days and Enhancing the

bioactivity of BCP

Chen et al.,2015

Acetone mix/dialysis35–125nm L-asparaginase Applying for the immobilization

of L-asparaginase

Zhang et al.,2007,2008

TABLE2.Materials for microcapsules preparation

Origin Type Materials Reference Natural polymers Lipids Ovolecithin,sphingomyelin Beamonte et al.,2013 Polysaccharide Alginate,chitosan,starch,agar Wang et al.,2015

protein Silk?broin,gelatin,albumin,?brin Liu et al.,2015

Semisynthetic polymers Cellulose derivative Sodium carboxymethyl cellulose,

cellulose acetate?bers KunDu et al.,2011 Dai et al.,2014

Synthetic polymers Biodegradation Poly lactone,polyanhydride Mao et al.,2012

Nonbiodegradation Polyamide,polyvinylchloridepolyacrylamide,

polystyrene Wang et al.,2014 Liang et al.,2015

4Q.LIU ET AL.

Microscopy Research and Technique

microcapsules were suitable for the localized delivery of plasmid DNA,combining low cytotoxicity and high transfection ef?ciency (Li et al.,2014).

Silk Fibroin Films

Silk ?broin ?lms can be easily fabricated by casting,methanol-treated,water-annealed,and LbL assembly.It is usually brittle,thus many studies prepare silk ?broin composite ?lms by blending with other poly-mers,including natural and synthetic polymers.

Solvent Casting.Silk ?broin composite ?lms can be simply prepared using solvent casting method.The silk ?broin–cellulose nano?ber composite ?lms could maintain high transparency and its mechanical strength was increased by 44%(Cho et al.,2014).Wang et al.fabricated silk ?broin/polyvinyl alcohol blend ?lms loaded with tetracycline and penicillin.The tetracycline-loaded ?lms showed no loss of activity over a 4-week period when stored at 4,25,and 378C,while only a 20%loss of activity at 608C,which was an important improvement in stability and potential delivery over tetracycline solution (Wang et al.,2010).In another study,Srivastava et al.successfully fabri-cated dextrose incorporated ?exible silk ?broin ?lms for wound dressing application.The ?exibility of silk ?broin ?lms was increased by addition of dextrose.The ?exibility of silk ?broin ?lms was capable of sup-porting L929?broblast cells growth and proliferation (Srivastava et al.,2015).Moreover,Seib et al.(2015)reported that crizotinib-loaded silk ?lms had an extended release pro?le and an improved response both in vitro and in vivo when compared to freely dif-fusible https://www.sodocs.net/doc/309243566.html,yer-by-Layer Assembly.LBL assembly approach has been applied in various ?elds of drug/gene delivery,scaffold fabrication for tissue engineer-ing,and biosensors (Mariana et al.,2015).For exam-ple,Jiang et al.developed a simple,ef?cient method to fabricate ultrathin silk ?broin ?lms by spin-assisted LBL assembly.The ultrathin ?lms were of outstanding mechanical properties and were observed with thick-nesses below 100nm,both in compressive and tensile modes.These outstanding properties were caused by the gradual development of the self-reinforcing micro-structure of highly crystalline b -sheet.But the con-?ned state of the silk ?broin backbones within ?lms was thinner than the lengths of the extended domains,which should be considered in the design of nanoscale ?lms and had potential application in microscale bio-devices and synthetic coatings for arti?cial skin (Jiang et al.,2007).In addition,silk ?broin composite mem-branes were prepared using LBL self-assembled tech-nology.The results showed that these ?lms were ?ne and uniform with the ?ber diameter from 400to 600nm,which had very large surface area and high porosity (Zhou et al.,2015).

Currently,silk ?broin ?lms can be prepared by numerous approaches such as solvent casting,methanol-treated,water-annealed.However,silk ?broin ?lms in the dry state usually become brittle.Hence,improvement of their mechanical properties via blending with other polymers is taken into account and has fueled broad interests in biomedical and clini-cal applications (as shown in Table 3).

T A B L E 3.S i l k ?b r o i n c o m p o s i t e ?l m s f o r c o n t r o l l e d r e l e a s e M a t e r i a l s

C a t e g o r i e s

P r e p a r a t i o n A d v a n t a g e s

C o m m o n a p p l i c a t i o n s

R e f e r e n c e s

S i l k ?b r o i n

S i l k ?b r o i n

C a s t i n g ;w a t e r -a n n e a l e d ;l a y e r -b y -l a y e r a s s e m b l y H i g h b i o c o m p a t i b i l i t y a n d

s l o w b i o d e g r a d a t i o n

D r u g /g e n e d e l i v e r y ,b i o m e d i c a l a n d c l i n i c a l a p p l i c a t i o n B y e t a l .,2005;J i a n g e t a l .,2007

S i l k ?b r o i n /n a t u r a l p o l y m e r c o m p o s i t e s

S i l k ?b r o i n /c h i t o s a n c o m p o s i t e m a t e r i a l s

F i l m s p r e p a r e d w i t h m e t h a n o l

E x c e l l e n t s t a b i l i t y ,c o m p r e s s i v e s t r e n g t h a n d c e l l u l a r c o m p a t i b i l i t y T h e t i s s u e s c a f f o l d o f c a r t i l a g e t i s s u e

D e v e c i e t a l .,2009;B h a r d w a j e t a l ,2012

S i l k ?b r o i n /c e l l u l o s e c o m p l e x m a t e r i a l s F i l m s p r e p a r e d w i t h s o l v e n t c a s t i n g S t r o n g h y d r o g e n b o n d i n t e r a c t i o n W o u n d d r e s s i n g

S e e t a l .,2014S i l k ?b r o i n /k e r a t i n c o m p o s i t e m a t e r i a l s

C a s t i n g i n p l a s t i c a n d d r y i n g

I m p r o v i n g t h e b i o c o m p a t i b i l i t y o f k e r a t i n N e r v e a n d l i g a m e n t r e g e n e r a t i o n

V a s c o n c e l o s e t a l .,2008

S i l k ?b r o i n /s y n t h e t i c p o l y m e r c o m p o s i t e s

S i l k ?b r o i n /p o l y e l e c t r o l y t e c o m p l e x e s C a s t i n g

E x c e l l e n t c e l l a d h e s i o n

B o n e r e p a i r a n d o t h e r s

B h a r d w a j e t a l .,2011;M a l a y e t a l .,2008

S i l k ?b r o i n /p o l y u r e t h a n e c o m p o s i t e s L a y e r -b y -l a y e r a s s e m b l y

H i g h m e c h a n i c a l s t r e n g t h D r u g d e l i v e r y

E l e a n o r e t a l .,2011;Z h o u e t a l .,2015S i l k ?b r o i n /p o l y c o m p o s i t e s C a s t i n g

I m p r o v i n g t h e b i o c o m p a t i b i l i t y C o n t r o l l e d r e l e a s e d r u g c a r r i e r s K u n d u e t a l .,2012;Y o u n g e t a l .,2007S i l k ?b r o i n /i n o r g a n i c c o m p o s i t e s S i l k ?b r o i n /s i l i c o n c o m p o s i t e s -

H i g h c o n t r o l l a b i l i t y

B o n e r e p a i r a n d r e g e n e r a t i o n -

S i l k ?b r o i n /h y d r o x y l a p a t i t e c o m p o s i t e s

M a t r i x -a s s i s t e d p u l s e d l a s e r e v a p o r a t i o n

E x c e l l e n t b i o c o m p a t i b i l i t y a n d m e c h a n i c a l p r o p e r t i e s

B o n e t i s s u e e n g i n e e r i n g

M i r o i u e t a l .,2010

PREPARATION OF SILK FIBROIN CARRIERS 5

Microscopy Research and Technique

Silk Fibroin Liposomes

Liposomes have been studied as a carrier for a few decades because they are nontoxic and nonimmuno-genic to human body and exhibit controlled release ability.Their release is affected by environmental stimuli such as changes in temperature and pH value. Temperature-responsive liposomes can be acquired by immobilizing a thermosensitive polymer on the sur-face.Whereas,the pH-responsive liposome can be acquired by immobilizing a titrable polymer on the surface.With the development of natural macromolec-ular material,silk?broin as the superior biomaterial can be used and prepared into liposomes.

Sangeeta et al.(2007)used freeze-drying method to prepare silk?broin coated liposomes.These liposomes encapsulated a receptor tyrosine kinase inhibitor,emo-din,working on Her2/neu overexpressing breast can-cer cells.The results showed that the delivery of liposomal emodin mediated by silk?broin was of higher ef?cacy in breast cancer cells,while the target-ing was related to the speci?city of emodin for Her2/ neu overexpressing cells(Dong et al.,2015).In another study,silk?broin liposomes were fabricated using evaporation method by Yuri et al.(2010).Their recombinant analogs did not lead to crucial perturba-tion,which was bene?cial to the further research of proteins toxicity to cells.

Silk Fibroin Microparticles

Silk?broin microparticles can be prepared using several methods,such as emulsion method(Cheerarot et al.,2015),self-assembly(Shi et al.,2011),and phase separation(Lammel et al.,2010).The microparticles can be coated with biological molecules to make them interact with a biological entity and provide a con-trolled release.Baimark et al.(2012)used water-in-oil emulsion solvent diffusion method to prepare silk ?broin microparticles.The prepared silk?broin micro-particles had spherical shape when adding Span80as the oil-soluble emulsi?er and could be controlled release via adjusting the degree of cross-linking.In another study,Lammel et https://www.sodocs.net/doc/309243566.html,ed phase separation method to fabricate silk?broin microparticles.The prepared process could be in?uenced by numerous parameters such as protein concentration,ionic strength,and pH value.The microparticles were capa-ble of loading small molecule model drugs,such as alcian blue,rhodamine B,and crystal violet,by simple absorption based on electrostatic interactions.The in vitro release behavior of the model drugs indicated that positively charged molecules were released in a more prolonged or sustained fashion.In addition,Shi et al.(2011)used self-assembly method to prepare silk ?broin microparticles with average diameter of 980nm.The microparticles were capable of loading drugs,such as FITC-BSA,RITC-Dextran,and RhB, with only23%FITC-BSA and34%RhB being released at their highest corresponding loading in50days.A simple and very mild processing method was devel-oped by Bessa et al.to fabricate silk?broin micropar-ticles.The microparticles could encapsulate human BMP-2and release it in a sustained way for a period of up to2weeks.When these microparticles were implanted in a rat ectopic model with5or12.5m g BMP-2,the bone density had a signi?cant increase (Bessa et al.,2010).In another study,Wu et al.pre-pared paclitaxel-loaded silk?broin nanoparticles for the purpose of enhancing antitumor ef?cacy.The in vivo antitumor ef?cacy was evaluated on gastric can-cer nude rat xenograft model.The results showed that the tumor weights were reduced and had no obvious toxicity,which indicated that paclitaxel-loaded silk ?broin nanoparticles were safe in vivo and had a potential application of clinical cancer treatment(Wu et al.,2013).

Although silk?broin nanoparticles could be a poten-tial future clinical application,the size distribution of silk?broin microparticles was inhomogeneous and less well suited to depot application(Kun et al.,2010). Hence,many approaches were developed to improve their size distribution,such as freeze-drying,rotary, and planetary ball milling which were summarized in Table4.

Silk Fibroin Microneedles Microneedles have been developed as micron-scale needles,which are prepared using numerous methods to administer molecules into the skin using a simple device.These microneedles present a promising tech-nology to deliver therapeutic compounds into the skin for a range of possible applications(Tsioris et al., 2012).You et https://www.sodocs.net/doc/309243566.html,ed reverse polydimethyl siloxane mold to prepare silk?broin microneedles.The dimen-sions of these silk?broin microneedles were about500 l m in length,200l m in diameter at the base,and5 l m in tip radii.The silk?broin microneedles were capable of transporting the loaded methylene blue or other compounds across the skin without giving rise to pain(You et al.,2011),which provided future promise for the delivery of a broad range of biomolecules including protein,peptides,vaccines,and nucleotides. In addition,Peter et https://www.sodocs.net/doc/309243566.html,ed a Clark-MXR-CPA-2010 laser micromachining instrument to prepare silk com-posite microneedles for loading vaccines.These micro-needles could stabilize subunit vaccines during room

TABLE4.Silk?broin-based microparticles for controlled release

Preparation Dimension Loaded molecule Key?ndings Reference Freeze drying<1l m-Appearance of absorption Nam et al.,2001 Emulsi?cation-diffusion167nm Fluorescent dyes Fabricating successfully

nanoparticle production

Yung et al.,2008

Rotary and planetary ball milling 200nm Ceramics Excellent nanoparticle

production

Rajkhowa et al.,2008

Self-assembly980nm Rhodamine B and crystal

violet Loading hydrophobic and

protein model drugs

Shi et al.,2011

Phase separation0.5–2l m Rhodamine B,alcian blue Loading small molecule

model drugs Wantanasiri et al.,2014

6Q.LIU ET AL.

Microscopy Research and Technique

temperature and enhance humoral immune responses when they were released over week-long(Peter et al., 2014).In another study,Lee et al.fabricated silk ?broin microneedles using thermally drawn micro-molds with methanol treatment.The results showed that methanol exposure time could effectively control the drug release rate from silk?broin microneedles (Lee et al.,2015).

Silk Fibroin Hydrogels

Hydrogels are emerged as three dimensionally cross-linked networks of hydrophilic polymers,which can imbibe large quantities of water or biological?u-ids.Hydrogels could be fabricated using water-soluble polymer or monomer,containing extensive chemical compositions and bulk physical properties(Guziewicz et al.,2011).Mariana et al.(2015)prepared silk?broin hydrogels with addition of several contents of ethanol, which could accelerate silk?broin gelation kinetics. The gelation process was affected by many parameters such as silk?broin concentration,temperature,pH value,and additives.The silk?broin hydrogels could be loaded with diclofenac sodium dissolved in ethanol and present more sustained drug release than hydro-gels loaded with the drug dissolved in water.In another study,silk?broin/nanohydroxyapatite com-posite hydrogels were fabricated using ethanol as gel-ling agent.The prepared hydrogels were favorable to new bone formation when their pore sizes were above 50l m.The metabolic and alkaline phosphatase activ-ities of osteoblastic cells were improved with the incor-poration of nanohydroxyapatite and silk?broin,which provided a promising future for bone tissue engineer-ing(Ribeiro et al.,2015).

CONCLUSIONS AND FUTURE PROSPECTS Silk?broin as a superior biomaterial has been pre-pared into many forms such as microspheres,?lms, hydrogel,liposomes,microneedles,and microcapsules under mild processing conditions.These formulations based on silk?broin are of high encapsulation ef?-ciency,long-term sustained release,and slow degrad-ability(Wang et al.,2007;Koh et al.,2015).However, these common production techniques usually create particles with a large size distribution due to the inho-mogeneous forces involved,which lead to the low bio-logical activity of loaded enzymes,cells,and growth factors.For instance,silk?broin microspheres pre-pared by spray drying,lipid templating,freeze-drying, and self-assembly caused a very broad size distribu-tion,which had an in?uence on the bioavailability (Breslauer et al.,2010).Thus,narrowing the particle size distribution has been indicated to be very impor-tant in controlling drug release kinetics and bioavaila-bility.Currently,micro?uidic approaches have been proven to be useful in the generation of monodisperse droplet suspensions,which are bene?cial to obtain nearly homogeneous microscale particles with micro-channel network and several functional units(Xu et al.,2009).Therefore,the carriers prepared by micro-?uidic approaches may acquire the relationship among the size distribution,controlled release and tissue regeneration.Moreover,the silk concentration can also alter their crystallinity and mechanical proper-ties,which would have in?uence on the release kinetics.Given these characteristics,micro?uidics is a promising choice to prepare silk?broin carriers for controlled release and biomedical applications.

REFERENCES

Alexander N,Mitropoulos Perotto G.2014.Synthesis of silk?broin micro-and submicron spheres using a co-?ow capillary device.Adv Mater26:1105–1110.

Vasconcelos A,Freddi G,Cavaco-Paulo A.2008.Biodegradable mate-rials based on silk?broin and keratin.Biomacromolecules9:1299–1305.

Asakura T,Ashida J,Yamane T.2001.A repeated b-turn structure in poly(Ala-Gly)as a model for silk I of Bombyx mori silk?broin stud-ied with two-dimensional spin-diffusion NMR under off magic angle spinning and rotational echo double resonance.J Mol Biol 306:291–305.

Baimark Y.2012.Morphology and thermal stability of cross-linked silk?broin microparticles prepared by the water-in-oil emulsion solvent diffusion method.J Chem Eng7:112–117.

Beamonte M,Roberto Lou B,Gracia M Osada J.2013.Sphingomyelin in high-density lipoproteins:Structural role and biological func-tion.Int J Mol Sci14:7716–7741.

Bessa PC,Balmayor ER,Hartinger J.2010.Silk Fibroin micropar-ticles as carriers for delivery of human recombinant bone morpho-genetic protein-2:In vitro and in vivo bioactivity.Tissue Eng16: 937–945.

Bhardwaj N,Kundu SC.2012.Chondrogenic differentiation of rat MSCs on porous scaffolds of silk?broin/chitosan blends.Biomateri-als33:2848–2857.

Bhardwaj N,Nguyen QT,Chen AC,Albert C.2011.Potential of3-D tissue constructs engineered from bovine chondrocytes/silk?broin-chitosan for in vitro cartilage tissue engineering.Biomaterials32: 5993–5781.

Bhrany AD,Lien CJ,Beckstead ND,Futran NH,Muni CM Giachelli BD.,Ratner.2008.Crosslinking of an oesophagusa cellular matrix tissue scaffold.J Tissue Eng Regen Med2:365–372.

Bini E,Knight DP,Kaplan DL.2004.Mapping domain structures in silks from insects and spiders related to protein assembly.J Mol Biol24:27–40.

Bourne PE,Weissig H.2003.Structural bioinformatics.Chichester: John Wiley&Sons Inc.12:649–655.

Breslauer DN,Muller SJ,Lee LP.2010.Generation of monodisperse silk microspheres prepared with micro?uidics.Biomacromolecules 11:643–664.

By H,Jaehyung P,Vassilis K.2005.Water-stable silk?lms with reduced b-sheet content.Adv Mater15:1241–1247.

Cao ZB,Chen X,Yao JR.2007.The preparation of regenerated silk ?broin microspheres.Soft Mater3:910–915.

Carl S,Bonde JS,Bulow L.2014.Characterization and assembly of a GFP-tagged cylindriform silk into hexameric complexes.Biopoly-mers101:378–390.

Cheerarot O,Baimark Y.2015.Biodegradable silk?broin/chitosan blend microparticles prepared by emulsi?cation-diffusion method.

E-Polymers15:67–74.

Chen L,Liu HL,Gu Y.2015.Lumbar interbody fusion with porous biphasic calcium phosphate enhanced by recombinant bone mor-phogenetic protein-2/silk?broin sustained-released microsphere: An experimental study on sheep model.J Mater Sci Mater Med26: 126–132.

Dai L,Shen Y,Li D.2014.Cellulose-graft-poly as a degradable drug delivery system.Cell Chem Technol48:237–245.

Dawson E,Mapili E,Erickson K.2008.Biomaterials for stem cell dif-ferentiation.Adv Drug Deliv Rev60:215–228.

Deveci SS,Basal G.2009.Preparation of PCM microcapsules by com-plex coacervation of silk?broin and chitosan.Colloid Polym Sci 287:1455–1467.

Dong YX,Dong P,Huang D,Mei LL,Xia YW,Wang ZH,Pan X,Li G, Wu CB.2015.Fabrication and characterization of silk?broin-coated liposomes for ocular drug delivery.Eur J Pharm Biopharm 91:82–90.

Du N,Yang Z,Liu XY,Li Y,Xu HY.2011.Structural origin of the strain hardening of spider silk.Adv Funct Mater21:772–778. Eleanor MP,David LK.2011.Silk?broin biomaterials for controlled release drug delivery.Drug Deliv8:797–811.

Farago S,Lucconi G,Perteghella S.2015.A dry powder formulation from silk?broin microspheres as a topical auto-gelling device.

Pharm Dev Technol10:1–10.

Gong XH,Liu HF,Ding XL,Liu ML,Li XM,Zheng LS.2014.Physio-logical pulsatile?ow culture conditions to generate functional

PREPARATION OF SILK FIBROIN CARRIERS7 Microscopy Research and Technique

endothelium on a sulfated silk?broin nano?brous scaffold.Bioma-terials35:4782–4791.

Guziewicz N,Best A,Ramirez BP,Kaplan DL.2011.Lyophilized silk ?broin hydrogels for the sustained local delivery of therapeutic monoclonal antibodies.Biomaterials32:2642–2650.

Tomoaki H,Tanimoto M,Shimabayashi S.2003.Change in secondary structure of silk?broin during preparation of its microspheres by spray-drying and exposure to humid atmosphere.J Colloid Interf Sci266:68–73.

Hofmann S,Foo CTWP,Rossetti F,Textor M,Novakovic G,Kaplan DL,Merkle HP,Meinel L.2006.Silk?broin as an organic polymer for controlled drug delivery.J Control Release111:219–227. Hosseinzadeh H,Atyabi F,Dinarvand R,Ostad SN.2012.Chitosan-pluronic nanoparticles as oral delivery of anticancer gemcitabine: Preparation and in vitro study.Int J Nanomed7:1851–1863. Holland TA,Mikos AG.2006.Biodegradable polymeric scaffolds.

Improvements in bone tissue engineering through controlled drug delivery.Adv Biochem Eng Biotechnol102:161–185.

Jiang CY,Wang XY,Gunawidjaja R,Lin YH,Gupta MK,Kaplan DL, Naik RR,Tsukruk VV.2007.Mechanical properties of robust ultra-thin silk?broin?lms.Adv Funct Mater17:2229–2237.

Jin HJ,Park V,Karageorgiou V,Kim UJ,Valluzzi R,Kaplan DL.

2005.Water-stable silk?lms with reduced beta-sheet content.Adv Funct Mater15:1241–1247.

Kakran M,Antipina MN.2014.Emulsion-based techniques for encapsulation in biomedicine food and personal care.Curr Opin Pharmacol18:47–55.

Kazemimostaghim M,Rajkhowa R,Tsuzuki T,Wang X.2013.Produc-tion of submicron silk particles by milling.Powder Technol241: 230–235.

Koh LD,Cheng Y,Teng CP.2015.Structures,mechanical properties and applications of silk?broin materials.Prog Polym Sci46:86–110.

Kun DJ,Chung YI,Kim YH,Tae G,Kundu SC.2010.Silk?broin nanoparticles for cellular uptake and control release.Int J Pharm 388:242–250.

Kundu J,Mohapatra R,Kundu SC.2011.Silk?broin/sodium car-boxymethyl cellulose bleed?lms for biotechnological Applications.

J Biomat Sci Polym22:519–539.

Kundu B,Rajkhowa R,Kundu SC,Wang XG.2013.Silk?broin bio-materials for tissue regenerations.Adv Drug Deliv Rev65:457–470.

Kundu B,Kurland NE,Bano S,Patra C,Engel FB,Yadavalli VK, Kundu SC.2014.Silk proteins for biomedical applications:Bioen-gineering perspectives.Prog Polym Sci29:251–267.

Lammel AS,Hu X,Park SH,Kaplan DL,Scheibel TR.2010.Control-ling silk?broin particle features for drug delivery.Biomaterials31: 4583–4591.

Lee JY,Park SH,Seo IH.2015.Rapid and repeatable fabrication of high A/R silk?broin microneedles using thermally-drawn micro-molds.Eur J Pharm Biopharm94:11–19.

Li LH,Puhl S,Meinel L.2014.Silk?broin layer-by-layer microcap-sules for localized gene delivery.Biomaterials35:7929–7939.

Liang K,Liu QQ.2015.Synthesis and characterization of hydroxyl-Functionalized polystyrene microspheres with monodispersity.

J Macromol Sci A52:436–443.

Liu HF,Fan HB,Cui YL,Chen YP,Yao KD,Goh JCH.2007a.Effects of the controlled-released basic?broblast growth factor from chitosan-gelatin microspheres on?broblasts cultured on a chitosan-gelatin scaffold.Biomacromolecules8:1446–1455.

Liu HF,Ge ZG,Wang Y,Toh SL,Sutthikhum V,Goh JCH.2007b.

Modi?cation of sericin-free silk?bers for ligament tissue engineer-ing application.J Biomed Mater Res B82B:129–138.

Liu HF,Fan HB,Wang Y,Toh SL,Goh JCH.2008a.The interaction between a combined knitted silk scaffold and microporous silk sponge with human mesenchymal stem cells for ligament tissue engineering.Biomaterials29:662–674.

Liu HF,Fan HB,Toh SL,Goh JCH.2008b.A comparison of rabbit mesenchymal stem cells and anterior cruciate ligament?broblasts responses on combined silk scaffolds.Biomaterials29:1443–1453. Liu HF,Li XM,Zhou G,Fan HB,Fan YB.2011a.Electrospun sul-fated silk?broin nano?brous scaffolds for vascular tissue engineer-ing.Biomaterials32:3784–3793.

Liu HF,Li X,Niu X,Zhou G,Li P,Fan Y.2011b.Improved hemocom-patibility and endothelialization of vascular grafts by covalent immobilization of sulfated silk?broin on poly(lactic-co-glycolic acid)scaffolds.Biomacromolecules12:2914–2924.

Liu L,Zhang XL,Liu X,Liu J,Lu G.2015.Biomineralization of sta-ble and monodisperse vaterite microspheres using silk nanopar-ticles.ACS Appl Mater Interfaces7:1735–1745.Malay€O,Yalcin D,Batig€u n A,Bayraktar O.2008.Characterization of silk?broin/hyaluronic acid polyelectrolyte complex?lms.

J Therm Anal Calorim3:749–755.

Mao SR,Guo CQ,Shi Y,Li LC.2012.Recent advances in polymeric microspheres for parenteral drug delivery-part.Expert Opin Drug Deliv9:1161–1176.

Mariana AM,Mahl CRA,Silva ME,Beppu MM.2015.Formation of silk?broin hydrogel and evaluation of its drug release pro?le.

J Appl Polym Sci8:185–191.

Mariana AM,Crouzier T,Rubner M,Beppu MM.2015.Factors con-trolling the deposition of silk?broin nano?brils during layer-by-layer assembly.Biomacromolecules16:97–104.

Meinel L,Kaplan DL.2012.Silk constructs for delivery of musculo-skeletal therapeutics.Adv Drug Deliv Rev64:1111–1122.

Miroiu FM,Socol G,Visan https://www.sodocs.net/doc/309243566.html,posite biocompatible hydroxylapatite-silk?broin coatings for medical implants obtained by matrix assisted pulsed laser evaporation.Mat Sci Eng B-Solid 169:151–158.

Mottaghitalab F,Farokhi M,Shokrgozar MA.2015.Silk?broin nano-particle as a novel drug delivery system.J Control Release206: 161–176.

Nam J,Park YH.2001.Morphology of regenerated silk?broin: Effects of freezing temperature,alcohol addition,and molecular weight.J Appl Polym Sci12:3008–3021.

Nam KG,Lee JS,Lee JH.2015.Growth,cell cycle progression,and morphology of3T3cells following?broin microsphere ingestion.

J Biomed Mater Res104:1325–1331.

Nayak S,Dey S,Kundu SC.2014.Silk sericin-alginate-chitosan microcapsules hepatocytesen capsulation for enhanced cellular functions.Int J Biol Macromol65:258–266.

Numata K,Subramanian B,Currie HA,Kaplan DL.2009.Bioengi-neered silk protein-based gene delivery systems.Biomaterials30: 5775–5784.

Numata K,Kaplan DL.2010.Silk-based delivery systems of bioactive molecules.Adv Drug Deliv Rev62:1497–1508.

Omenetto FG,Kaplan DL.2010.New opportunities for an ancient material.Science329:528–531.

Peter C,Muth D,Min YN.2014.Implantable silk composite micro-needles for programmable vaccine release kinetics and enhanced immunogenicity in transcutaneous immunization.Adv Healthcare Mater3:47–58.

Pritchard EM,Kaplan DL.2011.Silk?broin biomaterials for con-trolled release drug delivery.Expert Opin Drug Deliv8:797–811. Rajkhowa R,Wang L,Wang X.2008.Ultra-?ne silk powder prepara-tion through rotary and ball milling.Powder Technol185:87–95. Rammensee S Slotta T.,Scheibel.2008.Assembly mechanism of recombinant spider silk proteins.Proc Natl Acad Sci USA18:6590–6595.

Ribeiro M,Moraes MA,Beppu MM.2015.Development of silk ?broin/nanohydroxyapatite composite hydrogels for bone tissue engineering.Eur Polym J67:66–77.

Sangeeta KC,Andrea SG,Robyn R.2007.Silk?broin mediated deliv-ery of liposomal emodin to breast cancer cells.Int J Pharm341: 221–229.

Se YC,Min EL,Choi YG,Jin HJ.2014.Cellulose nano?ber reinforced silk?broin composite?lm with high transparency.Fiber Polym15: 215–219.

Seib FP,Coburn J,Konrad I.2015.Focal therapy of neuroblastoma using silk?lms to deliver kinase and chemotherapeutic agents in vivo.Acta Biomater20:32–38.

Shao Z,Vollrath F.2002.Surprising strength of silkworm silk.

Nature418:741–747.

Shchepelina O Drachuk IMK.,Gupta.2011.Silk-on-silk layer-by-layer microcapsules.Adv Mater23:4655–4660.

Shen G,Hu XY,Guan G,Wang L.2015.Surface modi?cation and characterization of silk?broin fabric produced by the layer-by-layer self-assembly of multilayer alginate/regenerated silk?broin.

PLoS One10:e0124811

Shi PJ,Goh JC.2011.Release and cellular acceptance of multiple drugs loaded silk?broin particles.Int J Pharm420:282–289. Sirichaisit J,Brookes VL,Young RJ,Vollrath F.2003.Analysis of structure/property relationships in silkworm(Bombyx mori)and spider dragline(Nephilaedulis)silks using Raman spectroscopy.

Biomacromolecules4:387–394.

Soppimath KS,Aminabhavi TM,Kulkarni AR,Rudzinski WE.2001.

Biodegradable polymeric nanoparticles as drug delivery devices.

J Control Release70:1–20.

Srisuwan Y,Srihanam P.2009.Preparation of silk?broin micro-spheres and its application to protein adsorption.J Macromol Sci A 46:521–525.

8Q.LIU ET AL.

Microscopy Research and Technique

Srivastava CM,Purwar R,Kannaujia R.2015.Flexible silk?broin ?lms for wound dressing.Fiber Polym16:1020–1030.

Tahamtan A,Tabarraei A,Moradi A,Dinarvand M.2014.Chitosan nanoparticles as a potential nonviral gene delivery for HPV-16E7 into mammalian cells.Artif Cells Nanomed Biotechnol15:1–7. Tharwat,Tadros.2013.Microcapsule encyclopedia of colloid and interface science.687–731.

Tsioris K,Raja WK,Pritchard EM,Panilaitis B,Kaplan DL, Omenetto FG.2012.Fabrication of silk microneedles for controlled-release drug delivery.Adv Funct Mater22:330–335.

Wang P,Pi B,Wang JN.2015.Preparation and properties of calcium sulfate bone cement incorporated with silk?broin and Sema3A-loaded chitosan microspheres.Front.Mater Sci9:51–65.

Wang W.Material useful for absorbing organic waste water,contains crop straw powder,ferric chloride,mica,hollow glass microsphere, ferric sulfate,ion-exchange resin,gallic acid,shell?sh powder,pol-ycarbonate,and https://www.sodocs.net/doc/309243566.html,104174389-A.2014-8-20.

Wang X,Wenk E,Matsumoto AL.2007.Silk microspheres for encap-sulation and controlled release.J Control Release117:360–370. Wang X,Wenk E,Castro G,Meinel L.2007.Silk coatings on PLGA and alginate microspheres for protein delivery.Biomaterials28:4161–4169. Wang X,Zhang XJ,Castellot I.2008.Controlled release from multi-layer silk biomaterial coatings to modulate vascular cell responses.

Biomaterials29:894–903.

Wang XQ,Yucel T,Lu Q,Hu X,Kaplan DL.2010.Silk nanospheres and microspheres from silk/PVA blend?lms for drug delivery.Bio-materials31:1025–1035.

Wantanasiri P,Ratanavaraporn J,Ratanavaraporn J,Yamdech R, Aramwit R.2014.Fabrication of silk sericin/alginate microparticles by electrohydrodynamic spraying technique for the controlled release of silk sericin.J Electrostat72:22–27.

Wang Y,Rudym DD,Walsh A,Abrahamsen L,Kim HJ,Kim HS, Kirker HC.2008.In vivo degradation of three-dimensional silk ?broin scaffolds.Biomaterials29:3415–3428.Wenk E,Merkle HP,Meinel L.2011.Silk?broin as a vehicle for drug delivery applications.J Control Release150:128–141.

Wu PY,Liu Q,Li RT,Wang J,Zhen X,Yue GF,Wang HY,Cui FB,Wu FL,Yang M.2013.Facile preparation of paclitaxel loaded silk ?broin nanoparticles for enhanced antitumor ef?cacy by locore-gional drug delivery.ACS Appl Mater Interfaces5:12638–12645. Xu Q,Hashimoto M,Dang TT,Hoare T,Anderson DG.2009.Prepara-tion of monodisperse biodegradable polymer microparticles using a micro-?uidic?ow focusing device for controlled drug delivery.

Small5:1575–1581.

Yucel T,Cebe P,Kaplan DL.2009.Vortex-induced silk?broin hydro-gels.Biophys J97:2044–2050.

You XQ,Chang JH,Ju BK,Park JJ.2011.Rapidly dissolving?broin microneedles for transdermal drug delivery.Mat Sci Eng C-Mater 31:1632–1636.

Young HP,Park YH.2007.The effect of casting solvent on the struc-tural characteristics and miscibility of regenerated silk?broin/ Poly(vinyl alcohol)blends.Fiber Polym8:579–585.

Yung SJ,Kim HS,Kim Y.2008.Fluorescent silk?broin nanoparticles prepared using a reverse microemulsion.Macromol Res16:604–608.

Yuri NA,Irina VP,Lyubov ID.2010.Interaction of recombinant ana-logs of spider silk proteins1F9and2E12with phospholipid mem-branes.Biochimica Biophysica Acta98:1172–1178.

Zhang YQ,Shen WD,Xiang RL,Zhang LJ,Gao WJ,Wang WB.2007.

Formation of silk?broin nanoparticles in water-miscible organic solvent and their characterization.J Nanopart Res9:885–900. Zhang YQ,Xiang RL,Yan HB,Chen XX.2008.Preparation of silk ?broin nanoparticles and their application to immobilization of L-asparaginase.Chem J Chinese U29:628–633.

Zhou WT,Huang HT,Du S,Huo YD,He JX,Cui SZ.2015.Removal of copper ions from aqueous solution by adsorptiononto novel polye-lectrolyte?lm-coated nano?brous silk?broin non-wovens.Appl Surf Sci345:169–174.

PREPARATION OF SILK FIBROIN CARRIERS9 Microscopy Research and Technique

华为 U8950D刷机,解锁,recovery,root软件+教程

华为U8950D刷机,解锁，recovery，root软件+教程 此文仅分享 1、解锁bootloader 对于大多数人来说对于命令都不怎么熟悉，所以我就介绍比较直观的解锁方式，解压华为U8950D（荣耀+）解锁脚本.zip后得到 1）去华为官网申请解锁码，申请好后进行第二步 2）安装手机驱动， 3）将手机进入fastboot模式，（长按音量下键+开机键），界面会一直卡在哪里，然后用数据线连接电脑， 4）双击“开始解锁”，出现 如果没有解锁过可以直接输入2然后敲回车，进入

按任意键，进入 此时左上角会有类似中间 这样的提示，继续按任意键进入以下界面：

在这里输入你的解锁码后敲回车。 5）等待重启后解锁完成。 6）如果要查询自己解锁没有进入fastboot后运行“开始解锁”选择1就可以看到自己的bootloader状态，LOCKED表示是锁的，UNLOCKED表示解锁. 方法一： 用方法一文件夹的文件（刷root不同） 2、刷入recovery 1）关机后继续进入fastboot模式，连接数据线 2）解压U8950D_OneKey_Recovery +root.zip，打开运行我刷recovery

进入： 按任意键继续 继续按任意键

按任意键： 完成后下面会提示 然后按任意键机子会重新启动。 此时刷入recovery成功·····拔掉数据线，为下一步做准备。 3、root···· 1）关机，长按音量上+开机键，进入recovery，由于没发截图，所以我只能这样给大家说，请各位细心操作， 2）进入recovery后，音量+-可以调上下，电源键是选择，选择高级功能，确定后会看到获取ROOT权限，按电源键选择， 3）重启手机，完成root 方法二：recovery、root。软件下载地址：(https://www.sodocs.net/doc/309243566.html,/share/link?shareid=106184&uk=1124315836)

山海经经典语录

山海经经典语录 导读：1、赤羽公子，你带着茉儿和芙儿赶快离开桃花坳。《山海经之赤影传说》 2、茉儿不就是由西向东，顺流而下的江流儿吗？《山海经之赤影传说》 3、只要赤羽哥愿意跟我在一起，芙儿便不再去东灵。《山海经之赤影传说》 4、传说玄女，可以唤醒四大神兽，得玄女者可得天下。《山海经之赤影传说》 5、你的心是最大的武器，找到七个守护的星宿。《山海经之赤影传说》 6、你知道唤醒青龙之后的后果吗？《山海经之赤影传说》 7、你看到这金盆里的水，这灾祸乃是因水而起啊。《山海经之赤影传说》 8、现身的不光有玄女，更有煞星。《山海经之赤影传说》 9、召唤朱雀，唤醒朱雀，只有朱雀的火焰才能烧尽滕蔓。《山海经之赤影传说》 10、我们召唤朱雀，是为了保护九黎的百姓。《山海经之赤影传说》 11、我真的是朱雀玄女吗？我要怎么样才能救我的族人呢？《山海经之赤影传说》

12、对于那些不怀好意的人，我，就意味着灾难。《山海经之赤影传说》 13、当玄女，可是桃花坳姑娘一生最大的荣耀。《山海经之赤影传说》 14、保护朱雀玄女，召唤朱雀神兽，就是我们的责任。《山海经之赤影传说》 15、你这个灾星，你换我桃花坳，你还我爹娘。《山海经之赤影传说》 16、你怎么可能是灾星呢，我会保护你的。《山海经之赤影传说》 17、混沌兽，不该对村民们下手，一定是有人暗中操纵。《山海经之赤影传说》 18、当一只自由自在的麻雀不好，那只凤凰没你想的那么容易当。《山海经之赤影传说》 19、唤醒朱雀，只有朱雀的火焰才能烧尽藤蔓，你的心是最强大武器。《山海经之赤影传说》 20、整个九黎，现在也只有你一个人有这个能力，去寻找朱雀玄女。《山海经之赤影传说》 21、玄女号令四方，拥有至高的权位，至高的荣耀，世人追求力量的野心是不会改变的。《山海经之赤影传说》 22、我来自桃花坳，我是一个江流儿，是桃花坳的族长老爹把我捡回家抚养长大的。《山海经之赤影传说》

MB860刷机完全教程

MB860刷机教程 免责声明 * 教程源于网络，并经过测试，但不代表所有机型通用； * 请务必保证电池电量在60％以上并保证刷机过程中电脑不会断电； * 刷机有风险，请仔细阅读相关的教程并自行承担刷机风险，一切后果自负； 刷机前置条件 1.手机电量充足，建议50%以上电量剩余 卡状态 手机连接电脑，默认充电状态，勿挂载存储卡、勿选择大容量存储，即电脑上无法使用存储卡；并确保存储卡容量400M剩余。 3.手机已root 可使用实用工具root；由于android版本多样化，root失败可参考下文手动root教程。 4.手机已解锁 判断解锁状态及解锁方法请参考下文。

MB860恢复官方兼救砖教程 刷机前准备： 1.电量保持在60%以上； 2.下载ME860工具包： 3.通过IMEI前6位（只能作参考）： ATT（美版）：353648，欧版：356381，亚太：356381 ，韩版：356455，国行：356451，台版：356451，港行：356451 版刷了美版底包，则不能刷入这里提供的ATT版本，参考知识库官方固件，刷入版。 国际版（除ATT外均是） 刷机工具：底包地址：版 底包地址：运行工具包内安装驱动； 6.运行工具包内安装MOTO官方刷机工具。 刷入底包： 1.手机关机，同时按音量上键+开机键进入RSD模式，再用USB线连接手机和电脑。 2.运行已安装好的RSD Lite刷机工具。 3.在RSD Lite界面，Filename栏选择工具包内的sbf文件，完成后点击start开始刷入：

当RSD刷机工具提示“Pleasemanually power up this phone”的时候表示已经刷底包完成；或者手机会自动重启，工具提示完成PASS。如果手机提示failed to boot，说明刷入底包出现异常，请认真确定自己的前置条件是否符合，并关机，正确按键（不一定会显示相关模式，按键正确即可），连接数据线，再进行刷机；最好更换官方底包。 WIPE并开机： 1.完全关机，同时按音量下键+电源键，直至手机显示fastboot字样。 2.按音量下键将显示各种模式，请将手机切换到Android Recovery模式，再按音量上键确认。 3.几秒后会显示三角形＋小机器人，点屏幕右下角的位置，或同时按下音量上键和下键，就进入了Recovery模式的菜单界面。 4.选择wipe cache partition清除用户缓存，触屏点击左下角OK确认。 5.选择wipe date/factory reset，进入二级菜单，选择delete all data，按左下角ok 确认。 6.选择reboot system now重启手机即可。 第一次进入系统较慢，请耐心等待。

山海经经典图片全集

本北方食人之兽，状如犬，传为海中神兽，状如马而有鳞，口中喷火，骘猛异常，食龙脑。其说多见于宋以后。

【原文】邽山，蒙水出焉，南流注于洋水，其中多黄贝①；蠃鱼，鱼身而鸟翼，音如鸳鸯，见则其邑大水。 【注释】①黄贝：据古人说是一种甲虫，肉如蝌蚪，但有头也有尾巴。 【译文】邽山，蒙水从这座山发源，向南流入洋水，水中有很多黄贝；还有一种蠃鱼，长着鱼的身子却有鸟的翅膀，发出的声音像鸳鸯鸟鸣叫，在哪个地方出现那里就会有水灾。

【原文】邽山，其上有兽焉，其状如牛，猬毛，名曰穷奇，音如嗥狗，是食人。 【译文】邽山，山上有一种野兽，形状像一般的牛，但全身长着刺猬毛，名称是穷奇，发出的声音如同狗叫，吃人。 或

【原文】阴山，有兽焉。其状如狸而白首，名曰天狗，其音如榴榴，可以御凶。 【译文】阴山，山中有一种野兽，形状像野猫却是白脑袋，名称是天狗，它发出的叫声与“榴榴”的读音相似，人饲养它可以辟凶邪之气。

【原文】章莪之山，无草木，多瑶、碧。所为甚怪。有兽焉，其状如赤豹，五尾一角，其音如击石，其名曰狰。 【译文】章莪山，山上没有花草树木，到处是瑶、碧一类的美玉。山里常常出现十分怪异的物象。山中有一种野兽，形状像赤豹，长着五条尾巴和一只角，发出的声音如同敲击石头的响声，名称是狰。

６【异兽】肥遗(鸟) 【原文】英山，有鸟焉，其状如鹑①，黄身而赤喙，其名曰肥遗，食之已疠②，可以杀虫。 【注释】①鹑：即“鹌鹑”的简称，是一种鸟，体形像小鸡，头小尾短，羽毛赤褐色，有黄白色条纹。雄性的鹌鹑好斗②疠：癞病，即麻风。 【译文】英山，山中有一种禽鸟，形状像一般的鹌鹑鸟，是黄身子而红嘴巴，名称是肥遗，人吃了它的肉就能治愈麻疯病，还能杀死体内寄生虫。

HTC_Sensation _G14 Z710e_解锁刷机全图文教程

HTC_Sensation (G14 Z710e) 解锁刷机全图文教程 目录 前言 (2) 第一篇：安装HTC SYNC (2) 第二篇：SHIP S-OFF到ENG S-OFF (2) 小白扫盲HBOOT,SPL (2) SHIP S-OFF的步骤： (3) 第三篇ROOT (7) ROOT方法： (7) 系统程序介绍 (7) 第四篇RECOVERY (12) RECOVERY步骤 (12) recovery里的各项菜单的功能中英文对照表 (13) 第五篇更新ROM (15) 参考资料及软件下载 (20) 本文参考资料来源 (20) 主要软件的下载 (20) 免责声明 (20)

前言 HTC_Sensation刷机方法教程许多网站都有介绍，对于有过成功刷机经历的人可以说是一目了然了，但对于初级菜鸟就有点狗咬刺猬无从下手了。本人本配合图示详细介绍了解锁与刷机过程，操作只要不出错，一般就会一次成功。 重要提示：过程较复杂，手机变砖概不负责，我刷入成功了。有好几个人手机已经变砖了，原因是刷机过程中手机会自动重启一次，接着自动刷。若在手机自动重启时，你看到黑屏，把电池扣了，估计就变砖了。 此办法目前仅限： HBOOT为1.17.0006、1.17.0008、1.17.0011、1.17.0012、1.18.0000这5个HBOOT 其它的机友暂请不要尝试，具体步骤，请严格谨慎进行！ 本文第一至第三篇主要讲HTC_Sensation的解锁，解锁后安装相关的软件后就可以删减系统中自己不喜欢的软件；建议一般的用户保留原装系统，必竟老婆是人家的好，系统是原装的好。本文第四至第五篇主要讲的是系统升级与更新，对于手机发烧友可以到网上下载自己喜欢的新版本系统进行安装，但必须注意的是一定要保证先要提升固伯版本，使其与系统相协调。 祝大家好运！ =================＝＝＝＝＝＝＝＝＝＝＝=======＝=====＝＝＝＝＝======== 第一篇：安装HTC SYNC 其实我们需要的是此软件里面的驱动程序。HTC SYNC下载地址：https://www.sodocs.net/doc/309243566.html,/managed-assets/support/software/htc-sync/setup_3.0.5617.exe如本软件更新，可以到https://www.sodocs.net/doc/309243566.html,/managed-assets/support/software/htc-sync 注意：安装过程中，所有进程都要是“成功”状态，否则会出问题。安装完成后，以及进行后续步骤的时候，一定要完全关闭HTC SYNC软件。 第二篇：SHIP S-OFF到ENG S-OFF 小白扫盲HBOOT,SPL 在HTC的Android 系统的系列手机中，HBOOT就是SPL。 SPL英文全称是Second Program Loader,“第二次装系统”，就是负责装载操作系统到RAM中。另外SPL还包括许多系统命令，如mtty中使用的命令等。SPL损坏了还可以用烧录器重写。SPL一般提供这几部分功能：检测手机硬件、寻找系统启动分区、启动操作系统为系统的基本维护提供操作界面，可以通过数据线与操作终端（如PC）建立连接，并接受和执行相应命令。它里面包含许多命令，像 r2sd,l,doctest(危险命令，他会擦除gsmdata)等。我们常说的三色屏就是由SPL 驱动的。检测SD卡，

《山海经》图像亡佚考

中国美术报/2018年/1月/1日/第017版 学术月刊 《山海经》图像亡佚考 山西博物院研究员渠传福 【编者按】2017年11月29日，上海博物馆与山西博物院共同举办的“山西博物院藏古代壁画艺术展”拉开帷幕，展览遴选了山西博物院珍藏的北朝和宋金元时期的12组89件墓葬壁画作品，其中大部分为首次公开展出。这是目前国内规模最大的一次壁画艺术原作特别展览。展览在开展前数月，就吸引了大批关注者。包括围绕特展组织的系列讲座，在开放预约一小时后，即已场场爆满。为了便于读者更好地理解中国古代壁画艺术和其所隐藏的历史文化内容，本刊特邀考古专家、山西博物院研究员渠传福就九原岗《升天图》与《山海经》的秘密问题进行讨论，以期为读者打开一扇通往国宝秘密的大门。 《山海经》研究作为一个文化热点课题，已经持续了相当长一段时期。众所周知，《山海经》有图有文，互为表里。历代注家和研究者对《山海经》古图的推测，大致可归纳为禹鼎说、地图说、壁画说和巫图说四种。有学者指出：“《山海经图》再现了中华民族童年的梦。神话是人类童年的梦，是人类走出混沌的第一声呐喊，是人类从自然走向文明所采摘的第一批果实。神话是民族生命力的源泉，是民族文化的根，是民族精神之所在。”更有人将其称为中华文化的“密码”。 历代学者的研究汗牛充栋，公认了一个令人悲哀的结论：到两晋后，郭璞和陶渊明所见的《山海经》古图已经全部亡佚。 其实未必，《山海经图》余绪尚存：郭璞之后200年，南朝梁张僧繇于6世纪初曾绘制《山海经图》十卷；唐初，裴孝源《贞观公私画史》中记录有《畏兽图》，应与《山海经图》有关；到唐末，《山海经图》和《大荒经图》已被张彦远《历代名画记》视为“古之秘画珍图”了。宋咸平二年（999）舒雅据张僧繇之残本重绘《山海经图》十卷。到南宋姚宽（1105—1162）《西溪丛语》说：“《山海经（图）》《大荒经（图）》，……此书今亡矣。”至此，《山海经》古图亡佚终于盖棺论定。 考据学派兴起，贡献丰富，唯于《山海经图》考证，虽着力不少，但多属闭门造车。目前所见明清时代的多种《山海经图》版本，以及日本学者的山海经《怪奇鸟兽图卷》大行其道，即其流风余韵。当代《山海经》学界许多人在此基础上，继续经营。更有不少画家浓墨重彩，描绘出若干套《山海经》异兽图集，也算别开生面。 有鉴于此，《山海经》学界的领军人物，喊出了“寻找失去的另一半”的口号，决心另辟蹊径，重新审视考古学成果，“从保存至今的《山海经》同时代的帛画、漆画、铜器上的针刻画入手，探寻已经失落了的山海经古图的人文特色与风貌。”从源头上追寻《山海经》古图像，以期续接《山海经图》的传承，可敬可佩。 然而，他们可能囿于古代先贤的定论和思维定式，似乎忘记质疑《山海经图》“两晋亡佚论”是否无懈可击。如此一套极大地影响中国文化的巨著尤其是图像，可能在动乱年代受到重创乃至毁灭，但有可能干净彻底地无影无踪吗？南北朝考古出土的一些古图像是否与《山海经图》有关？ 堪明中国社会对《山海经图》传承，同样是依据考古资料，如果说《山海经》学界是想从“亡佚前”追寻源头，本文则试图从“亡佚后”寻找其流变，想来应该可以殊途同归。九原岗《升天图》的出土，给了我们探讨这一课题的契机。 《山海经图》“两晋未亡论” 历代学者所谓《山海经图》于两晋亡佚的结论，应该指的是作为官私庋藏的“图书”（绢帛书、画）而言。而其他载体的《山海经》图像，譬如在殿宇壁画、墓葬壁画、砖石铭刻和各类艺

原创凤凰刷机-详细教程(图解)

凤凰刷机详细教程 —haottg 【刷机软件的选择】 刷机的方法常用的有三种：用NSS或者JAF修改手机CODE 后用在线更新升级、JAF刷机、凤凰刷机。 在线升级比较简单，只要修改CODE后用官方软件更新就行，这是比较多新手选用它的原因。但它不稳定，很容易出怪问题，例如：升级过程很顺利却不开机。数据损坏几率大，强刷都救不回的几率也就大了，所以不推荐。 JAF，曾经辉煌一时的软件，目前比较稳定的版本是1.98.62，但对新机型支持不太好，不支持刷C5-03。 凤凰，其实也是诺基亚的产品，功能极端强大。不过我们使用的破解版不能完全发挥出来，但刷机的功能是完整的。它刷机稳定、速度快、完美支持新机型，是刷机的首选。让新手望而却步的是看似复杂的刷机步骤，其实只要明白相关知识后再去看刷机教程，它也是很简单的，并且成砖几率可以说是零。 【刷机注意事项】 1、凤凰刷机软件是电脑软件，不要想着安装在手机上来进行刷机操作。 2、手机电池电量要充足，不需要满电，但最起码能保证通话10分钟的电量。 3、刷机不代表破解手机，刷已经固化免签或者深度破解软件的固件

才算是破解手机。 4、凤凰刷机不能降级刷，可以平刷以及升级刷。在手机待机状态下拨*#0000#可以查看手机版本号以及手机的RM。 5、C5-03的刷机固件是通用的，只要不是降级刷就行，不需要修改手机CODE。至于什么是CODE请别问我，自己百度去。 6、手机锁码默认是12345，刷机之后锁码不会改变，如果你在刷机前修改过，那么刷机之后会提示输入手机锁码。 7、内存卡取不取下来都无所谓，但我建议不要取下内存卡。 8、刷机不会失去保修，只要非人为硬件损坏，无论怎么刷机都是可以保修的，请放心。 9、只要是刷机所使用的固件没有问题，凤凰刷机可以说是零风险的，不必担心一些因素导致手机成砖后彻底报废这个问题。 10、刷机用的电脑一定不要开启任何杀毒软件以及防火墙，无论是个人电脑或者是网吧电脑，否则会造出不必要的麻烦。 12、网吧电脑一般来说都是有驱动安装限制的，请放弃使用网吧电脑进行刷机。 13、请不要问水货该怎么刷、港行又该怎么刷！刷机是不分行水货的！ 14、请不要问港行刷国行固件之后会不会不能用wifi！无论行水，C5-03都是可以用wifi的。 15、请不要问可不可以用jaf来刷C5-03，jaf是不支持刷C5-03的，当然你可以选择不信。 16、电脑联不联网都不影响刷机，请不要问没联网的电脑可不可以刷机！

《山海经》中奇花异草整理

《山海经》奇花异草整理 1建木-----沟通天地人神的桥梁 有木，其状如牛，引之有皮，若缨、黄蛇。其叶如罗，其实如栾，其木若蓲，其名曰建木。在窳西弱水上。 2.寻木---神木，长千里 《山海经·海外北经》：“寻木长千里，在拘缨南，生河上西北。”拘缨是传说中的古代北国名。 3.扶桑树----华夏古老的创世神话中的经典图腾 大荒之中，有山名曰孽摇【君页】羝。上有扶木，柱三百里，其叶如芥。有谷曰温源谷。汤谷上有扶木，一曰方至，一曰方出，皆载于乌。 4.柜格松---天下树木翘楚 《山海经》中记载，在西海之外、大荒之中有座山名叫方山，山上有棵青色大树，名叫柜格松，那也是太阳和月亮所出入的地方。也有说在昆仑山的玉门峰顶，有棵名叫柜格松的护山神木，高834仞，其松木迎风斜倚、俏立风雪，高盈百丈、摩天擦云，矗立天地之间，乃是天下树木翘楚，也是日月所出入之所。 5.文玉树-----传说中的五彩玉树 开明北有视肉、珠树、文玉树、玗琪树、不死树。凤皇、鸾鸟皆戴瞂。又有离朱、木禾、柏树、甘水、圣木曼兑 6.玗琪树-----生长红色玉石的树 开明北有视肉、珠树、文玉树、玗琪树、不死树。凤皇、鸾鸟皆戴瞂。又有离朱、木禾、柏树、甘水、圣木曼兑 7.不死树-----治疗筋骨疼痛、还是一种非常好的天然防腐剂，可以长 生不死 开明北有视肉、珠树、文玉树、玗琪树、不死树。凤皇、鸾鸟皆戴瞂。又有离朱、木禾、柏树、甘水、圣木曼兑

8.圣木曼兑----服食它的果实可以使人圣明智慧 开明北有视肉、珠树、文玉树、玗琪树、不死树。凤皇、鸾鸟皆戴瞂。又有离朱、木禾、柏树、甘水、圣木曼兑。 9.栾木-----枝、叶、果都可以制成长生不死的仙药 有云雨之山，有木名曰栾。禹攻云雨。有赤石焉生栾，黄本，赤枝，青叶，群帝焉取药。10.朱木----神话树名，也叫四味木 有盖山之国。有树，赤皮枝干，青叶，名曰朱木。 11.丹木-----可食用、能治好黄疸病、抵御火焰伤害 圶山，其上多丹木，员叶而赤茎，黄华而赤实，其味如饴，食之不饥。丹水出焉。《山海经·西次三经》崦嵫之山，其上多丹木，其叶如谷，其实大如瓜，赤符而黑理，食之已瘅，可以御火。《山海经·西次四经》 12.枫木----可以招魂 蚩尤所弃其桎梏，化为枫木。《山海经》山海经的这段记载非常模糊，蚩尤究竟为谁所桎梏，没有记载；蚩尤弃其桎梏是怎么回事？是逃脱了，还是被杀了，也没有记载。 所幸，在《尔雅·翼》中有记载可以补充：“旧说云黄帝杀蚩尤于黎山之上，掷其械于大荒之中，朱山之上，化为枫木之林，此貌夸父之杖化为邓林也”这里面就说的比较详细了，交代了是黄帝诛杀了蚩尤于黎山，然后将他的武器扔到了大荒之中，结果在朱山上他的武器化为一片枫木之林，作者推测这种说法与夸父“弃其杖，化为邓林”的说法是类似的。 13.帝女桑----赤帝女成仙之桑树 《山海经·中山经》：“又东五十里曰宣山……其上有桑焉，大五十尺，其枝四衢，其叶大尺馀，赤理黄华青柎，名曰帝女之桑。” 14.祝余---可以食用，有饱腹感 有草焉，其状如韭而青华，其名曰祝余，食之不饥。

HTC Touch Pro T7272 解锁、刷ROM、刷Radio详细教程(图解)

，电脑端右下角该标记由灰色变成绿色，即为连接，电脑端右下角该标记由灰色变成绿色，即为

，电脑端右下角该标记由灰色变成绿色，即为，电脑端右下角该标记由灰色变成绿色，即为连接结尾))，正常运行正常运行，，第一个界面如上图第一个界面如上图，，勾选里面那可勾选的框勾选里面那可勾选的框，，点击点击““next next””。此勾选过程此勾选过程分 步一样勾选点击一样勾选点击并点击并点击并点击““next next” ”。

4、正常情况下正常情况下，，出现如上提示出现如上提示””接下来接下来““，点击点击””update “。待。待显示进度条走完显示进度条走完100%100%， ，就完成了完成了刷机刷机刷机过程。 过程。

注：除普通数据线刷机法外还有注：除普通数据线刷机法外还有四色屏四色屏四色屏刷机刷机刷机法 法和存贮卡刷机法等，本人未试过，此处仅为整理他人经验。 *四色屏刷机法：同时按住开机键同时按住开机键((POWER POWER))和音量下键和音量下键((Voice Down Down))，用手写笔捅PP PP((Reset Reset))。出现四色出现四色屏 屏，用USB 线和电脑连接，等四色屏的线和电脑连接，等四色屏的““S erial erial””变成变成““USB USB” ”，进行刷机。*存贮卡刷机法：手机已经解锁；手机充电50%以上；下载的手机ROM 的nbh 部分直接改名为 RAPHIMG.NBH RAPHIMG.NBH((注意大写和中间的点号注意大写和中间的点号))，放入，放入存贮卡存贮卡存贮卡根目录下根目录下根目录下((储存卡格式储存卡格式须须为FAT32FAT32))；使手机手机进入三色屏， 进入三色屏，会自动变成白色刷机界面，按一下电源键放开，进入刷机状态，待中间的几个选项全部出现OK 后软启后软启。 。四、四、刷 刷R adio 步骤刷R adio 步骤同步骤同刷刷ROM 步骤步骤。 。*手机解锁时间约1分钟分钟，，刷R adio 约3分钟分钟，，刷ROM 约10-15分钟分钟。 。(整理自www.HTCPPC https://www.sodocs.net/doc/309243566.html,) .com)

中国经典古籍《山海经》

中国经典古籍《山海经》 《山海经》是中国先秦古籍，也是一部荒诞不经的奇书。该书作者不详，现代学者均认为成书并非一时，作者亦非一人。相关的信息，一起来看看。 《山海经》全书现存18篇，原共22篇约32650字。共藏山经5篇、海外经4篇、海内经5篇、大荒经4篇。《汉书·艺文志》作13篇，未把大荒经和海内经计算在内。山海经内容主要是民间传说中的地理知识，包括山川、道里、民族、物产、药物、祭祀、巫医等。保存了包括夸父逐日、女娲补天、精卫填海、大禹治水等不少脍炙人口的远古神话传说和寓言故事。 《山海经》具有非凡的文献价值，对中国古代历史、地理、文化、中外交通、民俗、神话等的研究，均有参考，其中的矿物记录，更是世界上最早的有关文献。 《山海经》版本复杂，现可见最早版本为晋郭璞《山海经传》。但《山海经》的书名《史记》便有提及，最早收录书目的是《汉书·艺文志》。至于其真正作者，前人有认为是禹、伯益、夷坚，经西汉刘向、刘歆编校，才形成传

世书籍，现多认为，具体成书年代及作者已无从确证。 《山海经》影响很大，也颇受国际汉学界重视，对于它的内容性质，古今学者有着不同的认识，如司马迁直言其内容余不敢言也，如鲁迅认为巫觋、方士之书。现大多数学者认为，《山海经》是一部早期有价值的地理著作。 内容介绍 《山海经》是一部记载中国古代神话、地理、植物、动物、矿物、物产、巫术、宗教、医药、民俗、民族的著作，反映的文化现象地负海涵、包罗万汇。除了保存着丰富的神话资料之外，还涉及到多种学术领域，例如：哲学、美学、宗教、历史、地理、天文、气象、医药、动物、植物、矿物、民俗学、民族学、地质学、海洋学、心理学、人类学等等，可谓汪洋宏肆，有如海日。在古代文化、科技和交通不发达的情况下，《山海经》是中国记载神话最多的一部奇书，也是一部地理知识方面的百科全书。 《山海经》全书现存18篇，据说原共22篇，约32650字。分为《山经》和《海经》两个大的部分，是一部富有神话传说的最古老的地理书。共藏《山经》5篇、《海外经》4

小米手机刷机教程

小米手机刷机教程

刷上MIUI的你，拿起手机，让我们一起来开始新手玩机之旅吧 :) 迈出第一步：挑选缤纷应用 各式应用程序将极大丰富手机的使用，因此首先来挑选对你味口的应用程序。点击桌面上的 [应用超市] ，MIUI已为你准备了多款热门精品应用，从便签记事、影音娱乐到聊天微博，选择你所喜欢的，来个一口气安装吧！（请在连接了WLAN无线网络的时候使用） 小贴士：要是看完推荐的应用还不过瘾，可以点击应用超市里面"好友推荐"的标签，里面是MIUI的用户们推荐的应用，可不要挑花了眼哦 > 你还可以从Google、安智、机锋等知名电子市场中下载更多应用。需要先安装相应市场（应用超市中有下载），然后便可以进去浏览更多应用。 > 想要安装从电脑上下载的应用该怎么办？先把下载的安装文件(后缀为apk)拷贝到手机sd卡上，然后拿起手机，点击桌面上的 [文件管理]，找到你所拷贝的文件，点击安装便可以了。 小贴士：不知道怎样拷贝文件到手机上？先用数据线把手机连接到电脑上，然后从手机顶部拉下通知栏，点击 [USB已连接]，然后点击 [开关] 即可。 拷贝完毕后，别忘记选择 [关闭开关] ，否则手机的文件管理会找不到sd卡的，因为它正被电脑所占用着。 现在我有好多应用程序了^__^ 稍微整理一下吧

安装成功的应用程序都会列在桌面上，但顺序可能不一定是你想要的，怎么办呢？ > 长按住一个应用，然后你就可以随意的移动它的位置了。如果移动到屏幕的边缘便可以滑动到下一个屏幕。 > 屏幕不够了，想增加一个屏幕？用两根手指在桌面上做收缩的手势，就可以进入屏幕缩略图模式。在缩略图模式中可以增加、删除屏幕，同样也可以长按一个屏幕来移动位置。 > 想删除一个应用该怎么办？长按住想要删除的应用，丢到顶部的垃圾篓里，点击确认便可。 第二步，接下来让我们来玩点个性~ 没有个性的手机是平淡无奇的，让我们来点个性！

山海经中的神话故事(精选10个)

山海经中的神话故事 山海经中的神话故事（一）： 【精卫填海】 女娃是炎帝最宠爱的小女儿，模样长得纤秀，性格却很倔强。姐妹们都喜欢打扮，惟独她酷爱体育，尤其水上运动，游泳划船，跳水冲浪，无一样不爱，无一样不精。某日清晨，风和日而，正是出游的好时光。女娃驾一叶扁舟，在碧波荡漾的东洋大海上遨游。海风微微地吹拂，海浪柔柔地起伏，带着小舟往大洋深处漂去。 年轻单纯的女孩，哪明白世道险恶，仍陶醉在蓝色的温柔里。霎时间，平静的大海变脸了，微笑的太阳不见了，轻轻海风变得比刀刃还锐利，软软海浪变得比铁锤还刚硬。女娃凭着高超技艺，劈波斩浪，左避右挡，与大海周旋。时间一分钟、一分钟地过去，一小时、一小时地过去，大海的浪涛越来越高，女娃的力气越来越弱。夜幕降临了，天地间一片黑暗，大概星星们闭上了眼睛，不忍目睹惨剧的发生：小舟被巨浪碾成了碎片，女娃被旋涡吸入了深渊，喧嚣的涛声盖住了女孩求救的呼叫，她永远也不能回去见她慈祥的父亲了。 几天过后，一只小鸟在女娃沉溺的水域破浪而出，花头颅、白嘴壳、红脚爪，样貌有点儿像乌鸦，它的名字叫精卫，是女娃不屈的冤魂所化就。 精卫栖身于布满拓木林的发鸠山上，它天天从发鸠山衔了小石子，或者小树枝，展翅高飞，直至东海，把石子或树枝投下去。日复一日，年复一年，不管是赤日炎炎还是雨雪霏霏，不死鸟精卫回翔在波涛汹涌、洁瀚无垠的大海上空，投下颗颗碎石、根根断枝，它不间断地叫着“精卫、精卫”，以激励自已的斗志，它要以锲而不舍的精神，将东海填平。 东海恼怒了，东海咆哮了，浪涛喧哗，白沫四溅：“你为什么要把我填平？你为什么恨我这么深？” 天空中传来精卫鸟仇恨的啼鸣：“因为你夺走了我年轻的生命，因为你还将夺走千千万万的年轻的生命。” “算了吧，小鸟儿！你就是填一千年，一万年，也填不平我呀！”东海用轰隆隆的大笑声来掩饰自己的窘态。 “我要填的！我要填的！我要一千万年、一万万年地填下去，哪怕填到世界末日，宇宙终结。”不死乌精卫悲啸着，飞翔着，从发鸠山至东海，循环往复，衔石投石，永无休止。 “精卫衔微木，将以填沧海”（东晋.陶渊明《读山海经》），精卫已升华为中华民族不屈的象征；孔子“知其不可而为之”（《论语.子路》），诸葛亮“鞠躬尽瘁，死而后已”（三国.诸葛亮《后出师表》），鲁迅“用这期望的盾，抗拒那空虚中的暗夜的袭来，虽然盾后面也依然是空虚中的暗夜”（鲁迅《期望》），无数志士为理想而在无望之中前仆后继，死不旋踵，他们都是精卫精神的继承者和体现者。 山海经中的神话故事（二）： 【夸父追日】 太阳每一天东升西落，丝毫没有在意北方大荒中的成都载天之山上，有一个巨人正全神贯注地观察着它。巨人用两条黄蛇作耳环，手里也把玩两条黄蛇，他是后土的孙儿，信的儿子，名字叫做夸父。 夸父身材高大，如山岳耸峙，却很天真，富于幻想，这两天，他在思考几个有关太阳的大问题：其一，太阳落入昧谷，黑夜便要降临；我热爱光明，憎恶黑暗，我要去追赶太阳，让他永驻天空。其二，太阳的圆脸上，沾染了不少黑斑；我期望太阳更明媚，更透亮，我要去追赶太阳，请他揩干净脸盘。其三，太阳在夏天喷吐了过多的光和热，到了冬天势必缺乏能量；我喜欢四季如春，不要酷暑也不要严寒，我要去追赶太阳，劝他平均分配热能。夸父想着想着，提起木杖，撒开两条长腿，就朝太阳追去。

番茄花园S10刷机解锁指南

番茄花园刷机解锁指南 注意事项： 刷机和会导致失去保修，并且三星支付永久作废，开机会提示红字，感叹号。如果你不接受，不要刷机。 常用组合按键（刷机必知） 1、强制重启方法：同时长按音量下键电源键；（任何情况下都可以强制重启） 2、进入挖煤模式（模式）：先拔掉线，关机状态下，同时按住音量下和，然后插线到电脑 3、进入刷机模式（模式）：关机状态下或同时长按音量下键电源键黑屏瞬间，同时长按音 量上键电源键键； 4、强行进入模式：连接电脑数据线，按住音量减小，，电源，秒左右 注意：刷机之前，港版系统一定要退出谷歌账户，如果是港版系统，需要刷一下最新国行底包。 一，解锁手机： 解锁会导致重置出厂，记得解锁之前备份资料！ 一共个锁 （锁，开发者选项里打开解锁，然后长按音量解锁） (重新激活锁定，设置里搜索重新激活锁定关闭) (锁，国行没有，听说正常使用天可以改变 状态，或者淘宝解锁) 1.发开发者选项里解锁打开 设置，关于手机，软件信息，编译编号连续点 次，返回一步，看到开发者选项进入，找到解 锁打开。如右图

2.关机状态下，同时按住音量下和不放，然后连接数据线到电脑，进入一个界面，左上角 是一个英文，下面是一个三角形的感叹号 这个界面提示要长按音量上键解锁，按住音量上键，就会进入解锁界面，然后会询问解锁，按一次音量增加，会重启重置出厂。（注意如果解锁过了进去就变成上锁界面，再执行就是上锁了） 3.进到系统之后再联网，再打开系统设置关于手机软件信息，狂按“编译编号”，出现开 发者选项，返回到设置，进入开发者选项，等待出现解锁开关出现，并且是灰色的不可操作的，就可以开始下面步骤了，再次回到，如下图就可以刷了 个，并且：(国行手机没有 ) 如果这个状态，还要解锁，请看下面方法：解决方法：重刷港版固件，进行开机设置时，

关于《山海经》古代经典的观后感

关于《山海经》古代经典的观后感 关于《山海经》古代经典的观后感1 在暑假里，爸爸送给了我一个礼物《山海经》。看到了这个书名，顿时我的脑海里浮现出无数的景像又是山又是海的，勾起了我很浓厚的兴趣。 但是当我看完了这本书我太震撼了。 《山海经》约成书于4000年前，它是一部先秦古籍。分为《山经》、《海经》两大类，共18卷。书中记载了古代的各种奇形怪状、拥有超人神通的奇灵怪兽500多个，内容包括了地理、神话、宗教等方面，也同时记载了许多具有神话传说性质的事情，在我看来称得上是一部神奇的百科全书。里面有的怪物可以治病，有的怪物一出现会发生天灾，还有的怪物名字就是自己的叫声，真是太不可思议了。 其中还有两种奇异的动物让我难忘：一种是比翼鸟，每只鸟只长了一只翅膀，所以必须得两只鸟并在一起飞才能飞的起来。还有一种叫相柳，它长了九个头，可以同时吃九座山上的食物，凡是相柳到过的地方，都被掘成沼泽和溪谷。 在海经第六卷，海内西经记载了让我感觉十分神奇的树。珠树：传说只生长珍珠的树。文玉树：当然就像字面上的意思就是

生长五彩美玉的树。玗琪树：生长红色玉石的树。还有最神奇的不死树：就是一种长生不死的树，人服食了它长生不老。 总结最后我觉得《山海经》这本书对我来说就像知识的山、知识的海，它以丰富和奇特的内容，为古往今来的人们所称道、所叹服。所以要想学习和探索中国的历史在我看来不可不读《山海经》。 关于《山海经》古代经典的观后感2 第一次知道《山海经》这本书是在那年热播的《轩辕剑》里，里面的角色“书香”是“书妖”原型就是《山海经》这本古籍，当时看到就觉得很惊奇，书香什么都知道，后来上网查了《山海经》才知道这是一本这样神奇的书，后来，就一直想看，但一直没机会。又一次，一个偶然的机会，看了这本书。 全书共分有四个部分，依次是《藏山经》、《海外经》、《海内经》和《大荒经》。《藏山经》主要记载山川地理，动植物和矿物等的分布情况;《海经》中的海外经主要记载海外各国的奇异风貌;《海内经》主要记载海内的神奇事物;《荒经》主要记载了与黄帝、女娲、大禹等有关的许多重要神话资料。文字晦涩难懂，记录一个事物往往只用两行字，却要你用很久的时间思考这到底是什么东西，有的时候甚至五个字中三个都不认识。不过也就是那个时候我明白了，《山海经》也就只是《山海经》，没有人可以证实它是否存在，一切都是荒谬的，这种神物本不应该存

HTC radar解锁刷机详细教程,看完这个贴,如果还不会刷那你就来揍我吧!

但我还是担心有些同学看的不是很懂，所以今天再发一帖，尽量详细，详尽，当然如果有觉得D.F.T的ROM不稳定想换XDA的ROM的，也可以试试！如果同学们刷机有任何问题， 好了，闲话不多说了，开始！ 一：准备工作 一部雷达（radar/C110e/Omega） 目前支持的MID名单："PI06****”, “PI061100, “PI061000" 老规矩，回复满50取消回复可见 手机关机----同时按住音量键上和音量键下（保持不要松）----按开机电源键----直到出现USB HOST 模式----上面会出现一行数字，那个就是你机器的MID

如图 【这个要看清楚了，****不是指任意符号，而是手机上就显示****，一个数字不多一个不少，也有部分PI06100的机器可以解锁，但要求是欧版，如果分不清楚你的radar是欧版和亚太版，可以尝试刷下SPL，亚太版会提示错误，不过不要担心，不会变砖，重启即可】 如果你的手机MID不属于上述之列，那你需要用Y线金卡先刷SPL， （Y线制作和金卡比较麻烦，其实我建议如果真要Y线和金卡刷，Y线可以直接在淘宝买，自己做的太麻烦；至于金卡，对卡有限制，而且后面刷SPL的时候对读卡器也有限制，比Y线更麻烦。所以不喜欢折腾的同学还是等待以后所以MID都支持以后再刷机吧） 当然如果你MID在上述名单，或者已经决定用Y线金卡刷机，那么请接着往下看！ 先下载几个必要的程序 HSPL（里面含SPL和HSPL，刷机必备）【20.49M】（下载地址： radar第三方ROM （此ROM及radio来自XDA，用的是ROM刷机工具，有别于D.F.T）【418.75M】（下

山海经经典句子

山海经经典句子 1、天吴，八首八面，虎身，八足八尾，系青黄色，吐云雾，司水。 2、青丘之山：有兽焉，其状如狐而九尾，其音如婴儿，能食人，食者不蛊。 3、西北海之外，赤水之北，有章尾山。有神，人面蛇身而赤，身长千里，直目正乘，其瞑乃晦，其视乃明，不食不寝不息，风雨是谒。是烛九阴，是谓烛龙。 4、英鞮之山，涴水出焉，而北流注于陵羊之泽。是多冉遗之鱼，鱼身蛇首六足，其目如马耳，食之使人不眯，可以御凶。 5、天山，有神焉，其状如黄囊，赤如丹火，六足四翼，浑敦无面目，是识歌舞，实为帝江也。 6、流沙之滨，赤水之后，黑水之前，有大山，名曰昆仑之丘。 7、其状如狸而白首，名曰天狗，其音如榴榴，可以御凶。 8、有兽焉，其状如彘而人面。黄身而赤尾，其名曰合囗，其音如婴儿，是兽也，食人，亦食虫蛇，见则天下大水。 9、东海中有流波山，入海七千里。其上有兽，状如牛，苍身而无角，一足，出入水则必风雨，其光如日月，其声如雷，其名曰夔。黄帝得之，以其皮为鼓，橛以雷兽之骨，声闻五百里，以威天下。 10、北海鲛人的歌声迷惑人心。蠃鱼的啼叫带来水灾。鵸鵌以鸟鸣编制噩梦。捂紧耳朵，别听异兽的呓语。 11、钱来之山，其上多松，其下多洗石。有兽焉，其状如羊而马尾，

名曰羬羊，其脂可以已腊。 12、昆仑之丘，是实惟帝之下都。神陆吾司之。 13、再往西三百五十里，有座山名叫天帝山，山上是茂密的棕树和楠木树，山下主要生长茅草和蕙草。山中有一种野兽，形状像普通的狗，名叫谿边，将这种兽皮铺垫在身下，身体可以不受邪气的侵害。14、又西三百五十里曰天山，多金玉，有青雄黄，英水出焉，而西南流注于汤谷。有神鸟，其状如黄囊，赤如丹火，六足四翼，浑敦无面目，是识歌舞，实惟帝江也。 15、有草焉，其状如韭而青华，其名曰祝余，食之不饥。 16、又北二百里，曰少咸之山，无草木，多青碧。有兽焉，其状如牛，而赤身、人面、马足，名曰窫窳，其音如婴儿，是食人。敦水出焉，东流注于雁门之水，其中多魳魳之鱼。食之杀人。 17、君子国在其北，衣冠带剑，食兽，使二大虎在旁，其人好让不争。 18、邽山，蒙水出焉，南流注于洋水，其中多黄贝；蠃鱼，鱼身而鸟翼，音如鸳鸯，见则其邑大水。 19、西王母梯几而戴胜。其南有三青鸟，为西王母取食。在昆仑虚北。 20、凤凰栖息梧桐，五年方成神木，浴火重生，再十年，神木成圣。 21、大荒之中，有山名曰丰沮玉门，日月所入。有灵山，巫咸、巫即、巫盼、巫彭、巫姑、巫真、巫礼、巫抵、巫谢、巫罗，十巫从此升降，百药爰在。 22、混沌长毛四足，如犬，有腹无五脏。抵触善人，凭依恶人。应声虫居于…见则能霸天下。契俞兽中最大者，龙头马尾虎爪，长四百尺。

番茄花园Note9解锁,刷机教程

番茄花园Note9解锁，刷机教程 温馨提示： 刷入第三方包会触发Knox 0变成1，会影响保修，Samsung Pay和安全文件夹将永久无法使用。开机会提示非原装，请自行斟酌决定，本人概不负责。刷机有风险，刷机之前请看下载根目录的教程！。 首次刷机之前的准备工作： 刷机工具odin3 （网盘根目录） TWRP和ROOT （网盘根目录） 驱动程序（网盘根目录） 一，解锁CROM和BL 1.解锁（已解锁忽略） 进入开发者选项，打开OEM解锁开关 进入设置，关于手机，软件信息，找到 编译编号，连续点8次，然后返回上一 页，进入开发者选项，里面找到OEM 解锁打开就行。

2.关闭重新激活锁定（已解锁忽略） 进入设置，生物识别和安全性，查找我 的手机，登录三星账号，按照提示关闭 就行了，如果你从来没打开过忽略就行。 3.最后进入download确认左上角信息（2个OFF就行） 进入download方法看最下面一段 4.港版系统第一次需要刷国行四件套 如果你是港版系统，第一次需要下载国 行四件套刷成国行，刷港版系统之前， 需要退出谷歌账户（如果有登录的话）， download显示2个off

二，开始刷机（卡刷线刷2选1） A：线刷方法（推荐） 下载线刷版的rar压缩包，解压，打 开里面的odin，BL AP CP CSC分别载入 4个解压目录下的文件。如下图 手机进入download模式，连接数据线 到电脑，点start开始刷机就行了 进入download方法看最下面一段 小提示：2个CSC文件如何选？ CSC_WIPE_xxx，表示刷机会格式化data，如果你现在是官方，需要选这个CSC_WIPE 开头的载入CSC CSC_xxx表示，刷机可选删除或者保留数据，需要你是第三方，CSC选CSC_xxx B：卡刷步骤 1.刷入TWRP并且格式化data分区（具体刷入方法看下面） TWRP见每个版本的子目录下，已经有TWRP的可以忽略，番茄ROM可以忽略这个步骤 2.复制刷机包到内置储存刷机 内置储存读写速度快，不会出现刷机卡第一屏，并且TWRP可以连接电脑拷贝刷机包，如果你是官方系统，需要先格式化data再拷贝刷机包

《山海经》瑞兽整理

《山海经》中的瑞兽整理 1.天犬——安享太平 天犬，状如狗，身有豹纹，长尾，正在四处张望。 天犬，长得像狸猫，常发出喵喵的叫声，人饲养它便可以家宅安宁、万事太平。传说，在秦襄公的时候，白鹿原上原来有一个叫狗枷堡的村子，曾有天狗降临。只要有贼，天狗便狂吠而保护整个村子。天狗有食蛇的本领，它也被看作可以抵御带来凶灾的奇兽。 2.龙龟——衣锦荣归 龙首龟身。 龙龟，为古代神龙所生之子，背负河图洛书，揭显天地之数；上通天文，下知地理，中和人世。龙龟是纯阳之神兽，能避邪、制煞、化冲、解厄、镇宅、招财、聚财，是权力和长寿的象征。此外由于“龙龟”与“荣归”同音，龙龟也代表“衣锦还乡，荣归故里”一说。 3.三青鸟——好事连连 三青鸟，长有头冠，尾羽很长，从空中飞来。 三青鸟，色泽亮丽、体态轻盈，传说为女神西王母的使者，专门为西王母取食，人间不得见，唯有在蓬莱仙山可以见，但是蓬莱无路，只有靠青鸟传信。传说西王母驾临前，总有三青鸟先来报信。因此，三青鸟被当作传递信息的神鸟，后人将它视为传递幸福佳音的使者。 4.当康——五谷丰登 当康为猪形兽，满嘴猪牙。它的前身和头部微低，似在示威。 钦山中栖息着一种野兽，其外形像猪，长着大獠牙，名字叫当康。虽然它的样子不太好看，却是一种瑞兽。当天下要获得丰收的时候，当康就从山中出来叫，告诉人们丰收了。所以据《神异经》记载，南方有一种奇兽，样子像鹿，却长着猪头和长长的獠牙，能够满足人们祈求五谷丰登的愿望，可能就是当康。 5.蛮蛮——比翼双飞 蛮蛮就是比翼鸟，被视为成双成对的吉祥之鸟。

崇吾山中生活着一种像野鸭子的鸟，它只长了一只翅膀和一只眼睛，因此无法独自飞翔，需要两只鸟结对比翼齐飞，它的名称是蛮蛮。蛮蛮就是比翼鸟，古人视成双成对为吉祥，因此“不比不飞，不比不行”的观念成为中国吉祥文化的重要内容，夫妻、恋人往往有“在天愿做比翼鸟”的誓言。 6.鹿蜀——子孙满堂 鹿蜀站立在杻阳山上，马的样子却披着虎纹，回首扬尾。 杻阳山有一种瑞兽，名叫鹿蜀。鹿蜀的外形像马，白头、红尾，通身是老虎的斑纹。鹿蜀的鸣叫如同人在唱歌。古人认为披上鹿蜀的皮毛，就可以子孙满堂。郭璞在《图赞》里说：“鹿蜀之兽，马质虎文。骧首吟鸣，矫足腾群。佩其皮毛，子孙如云。” 7.白狼——政清人和 只有王者行仁政、知明哲，德至鸟兽时，白狼才会出现。 在中国古代，白狼是一种祥瑞的动物，它的出现往往和圣人及改朝换代联系在一起。《帝王世纪》记载：“汤得天下，有神獐、白狼衔钩入殿朝。”孟子说五百年必有圣人出，因此当商朝取代夏朝的时候，祥瑞的白狼就出现了，预示着商朝替换夏朝统治天下。唐朝编成的《艺文类聚》解释白狼说：“白狼，王者仁德明哲则见。”就是说白狼只会出现在国君具备仁德的时代，乱世是绝不会出现的。《山海经》的注者郭璞也称颂，唯有道德的人才能见到白狼。 8.鹿——长寿昌盛 鹿是长寿的仙兽，传说一千年为苍鹿，两千年为玄鹿，民间传说中的老寿星总是与鹿相联系。鹿乃纯阳之物，生命力极强，动作矫健，素有“草上飞”之称，即使腿骨折断，不需治疗也能自然愈合。“鹿”字又与“福、禄、寿”中的“禄”字同音，因此它常被用来表示长寿和繁荣昌盛。 9.寓鸟——世界和平 寓鸟外形与老鼠相似，长着宽宽的翅膀。 虢山中的禽鸟大多是寓鸟，它的形状与一般的老鼠相似，还长着鸟一样的翅膀，发出的声音就像羊叫。饲养它可以避除邪气，不受兵戈之苦。《中国神话·奇兽·古物总集》称：“寓鸟有一对宽宽的翅膀，身形似鼠，鸣声如羊。此鸟具有预报兵情的特异功能；豢养在家，可预防血光之灾、兵戈之害。” 10.毕方--赐予世人光明与温暖 古籍有云：“毕方形如仙鹤，鸣声迤逦”。《山海经》中的毕方是黄帝卫车旁的神鸟，可