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To explore methods and clinical effects of selective U-shaped osteotomy of lateral tibial condyle in treating collapse and comminuted fracture of lateral tibial plateau.From January 2014 to October 2019, 15 patients with collapse and comminuted fracture of lateral tibial plateau were treated by selective U-shaped osteotomy of lateral tibial condyle, including 9 males and 6 females. The age of patients ranged from 25 to 70 years old, with an average age of (38.5±7.7) years old. According to ABC classification of condyle fracture of tibial plateau lateral, there were 2 cases of type A, 6 cases of type B, 4 cases of type BC and 3 cases of type C. Five patients were combined with medial plateau fracture, 8 patients were combined with left knee fracture and 7 patients of right knee fracture. The time of treatment after injury ranged from 1 day to 14 days with an average of (3.4±1.2) days. CT of all patients showed that lateral tibial plateau collapsed more than 2 mm, more than 2 pieces of bones were crushed and broken, and lateral tibial condyle cortex was intact. At follow-up of 12 months after operation, Rasmussen's anatomical grading system was used to evaluate fracture reduction. Rasmussen's functional grading system were used to evaluate knee joint function.Selective U-shaped osteotomy was successfully complated in 15 patients at one time, and operation time ranged from 55 to 110 min, with an average time of (85.6±20.0) min. The lateral plateau operation ranged from 20 to 60 min with an average time of(30.5±10.5) min. All patients were followed up for 12 to 24 months with an average of (14.6±2.5) months. Fracture healing time was 12 to 24 weeks, with an average of (13.6±3.6) weeks. At follow-up 12 months after operation, by Rasmussen's grading system, anatomical score of knee joint ranged from 14 to 18 points, with an average score of (17.5±0.3) points, of which 13 cases were excellent and 2 cases were good. The functional score ranged from 13 points to 30 points, with an average score of (26.8±2.5) points. Among them, 12 cases were excellent, 1 case was good, 2 cases were fair. Two patients suffered 2 mm and 4 mm loss of lateral tibial plateau, 1 case of knee joint 5 ° valgus, 1 case of stiff joints (10 ° to 100 °). No common peroneal nerve injury, important vascular injury, postoperative infection, internal fixation failure and other serious complications was found.The use of selective lateral tibial condyle "U"- shaped osteotomy approach is an effective and reliable method to treat the collapse and comminuted fracture of the lateral tibial plateau. It has the advantages of simple surgical incision, direct fracture exposure, accurate repositioning and fixation, short operation time and few complications.

Calcium polyacrylate/alginate (CPA/A) hybrid polymer microspheres with the imprinting of bovine serum albumin (BSA) were prepared via Ca2+ crosslinking of sodium polyacrylate (SPA), BSA and sodium alginate (SA) in inverse suspension. The specific rebinding properties of BSA imprinted hydrogel microspheres (CPA/A MIPMs) were investigated by controlling pH value from the viewpoint of adjusting the process of gelling, removing template and rebinding. The optimized pH values in gelling, removing template and rebinding process was 4.1, 8.3 and 4.2, respectively.

Calcium phosphate/polyacrylate/alginate hybrid polymer microspheres with bovine serum albumin (BSA) embedded and coated on the surface were prepared with (NH4)2HPO4, sodium polyacrylate (SPA) and sodium alginate (SA) via Ca2+ crosslinking in inverse suspension. Rebinding behaviors of the microspheres were evaluated. The factors influencing the imprinting efficiency (IE) of imprinted microspheres were also studied, including the concentration of CaCl2, template content and pH values in rebinding solutions. Selectivity tests showed that the imprinted microspheres exhibited good recognition property for the template protein.

Magnesium alloys are potential in biomedical materials, but their corrosion rates are high in human body fluid so that they can not be used enough time. Therefore, to improve the anti-corrosion property of the alloys is important for their application in biomedical materials. In the work, a new coating was prepared from poly (3-hydroxybutyrate) (PHB), a biopolymer used as a sustained release carrier of drugs. The effect of the PHB coating was studied by electrochemical tests, scanning electron microscope (SEM) and weight loss corresponding with the corrosion time. The result showed the sample coating PHB have a higher corrosion potential than one before coating PHB, that is, PHB coating is effective for improvement of corrosion resistant of the alloy. Moreover, the corrosion state on the surface is even by SEM, implied the corrosion rate of the PHB coating is divinable and controllable. Whereby, PHB should be suitable for anti-corrosion of magnesium alloys applying in biomedical materials.

Summary: A method is presented that can produce ordered macroporous alginate membranes and microspheres with a relatively narrow pore size distribution as compared to other existing porous alginate matrix generating procedures. The calcium alginate membranes and microspheres obtained were multiporous, and the pore size could readily be adjusted in the range of 1 to 10 µm by selecting appropriate initial emulsion parameters. A preliminary set of experiments was used to determine a range of conditions suitable for the formation of stable O/W emulsions consinsting of the alginate solution, liquid paraffin and surfactants. The obtained emulsions were characterized by size distribution measurements and optical microscopy. The emulsions were used to form alginate membranes and microspheres with diameters of 150–500 µm. Subsequent work was performed to remove liquid paraffin droplets, the dispersed phase of the emulsion which acted as template in the pore-generating process, from the final alginate membranes and microspheres. The states of templates in hydrogels and the effect of template removal on the calcium alginate structures were thoroughly investigated. Porous alginate membranes prepared via emulsion templating method.

The hepatitis B virus X (HBx) protein is thought to be implicated in the development of human hepatocellular carcinoma (HCC), but its exact function remains controversial. To investigate whether the expression of the HBx gene alone can induce HCC on an inbred C57BL/6 strain that displays a lower spontaneous rate of liver cancer, and to determine if HBx transgenic mice are more susceptible to the effects of hepatocarcinogens, C57-TgN (HBx) X transgenic mice were bred with normal C57BL/6 mice strain. The F1 mice (about 50% HBx positive and 50% HBx negative) were treated with a single dose of diethylnitrosamine (DEN) at 7 days of age, or were untreated. Mice were killed at appropriate time points and were analyzed for histological change in the liver. The expression of HBx protein were examined by using immunohistochemical staining. Glycogen storage foci were examined by using periodic acid-Schiff (PAS) staining. In HBx transgenic mice untreated with DEN, HBx expression and glycogen storage foci were always observed in the liver after 8 weeks, but not obvious histological pathologic changes. Histological examination of liver tissue confirmed that DEN-treated HBx mice developed approximately twice as many focal lesions of basophilic hepatocytes as treated wild-type littermates. Hepatocellular adenomas and carcinomas were also more frequent in DEN-treated HBx-positive than HBx-negative mice. Taken together, our results suggest that HBx gene expression alone is not sufficient for carcinogenesis, but may act as a promoter for malignant transformation.

The present work has generated transgenic mice with a hybrid gene construct consisting of genomic sequences encodinghuman erythropoietin (hEPO) and governed by regulatory sequences of mousewhey acidic protein (mWAP). The construct proved effective by transient expression in lactating animal. After introducing hybrid gene construct into single-cell embryo via pronuclear microinjection, surviving embryo are reimplanted into pseudopregnant foster mother mouse. 58 mice of 86 generation zero mice obtained were identified to be positive by PCR-Southern blot and genomic DNA Southern blot methods. The integration rate is 67%.hEPO was expressed in the milk of 16 mice of 39 mice measured byhEPO ELISA kit The expression level gets over 15 μg/mL.

The structure and characteristics of AOT (aerosol OT, sodium bis (2-ethylhexyl) sulfosuccinate)-water-isooctane reverse micellar micro-reactors have been studied by means of electrical conductivity, infrared spectra (IR), nuclear magnetic resonance (NMR) and differential scanning calorimetry (DSC). The influences of temperature and organic additive are discussed. Nanometre-sized Fe3O4 and ZnS particles are prepared by reaction in the microreactors and the properties of the obtained nanoparticles are studied with the help of a transmission electron microscope (TEM) and ultraviolet (UV) spectra, respectively.

The swelling behavior of the complex film formed from pectin and chitosan was studied. The swelling degree increased sharply in the case of pH less than 2 and larger than 7. The swelling mechanisms of the complex film in acidic and alkaline medium are discussed, respectively. Moreover, the swelling degree could be modulated by means of the weight ratio of pectin to chitosan and the original weight concentration of pectin aqueous solution. © 1996 John Wiley & Sons, Inc.

A quick and simple approach for the measurement of H2S solubility in alkanolamine solutions (AAS), i.e. the electrode method, was recently reported by Rochelle et al. (1987). This is an ingenious new approach based upon the relationship of activities of hydrogen and sulfide ions with the equilibrium vapor pressure of H2S in AAS. In the present work the new method was experimentally studied and compared with the Rochelle's report and the conventional method. Additionally the effect of existence of CO2 in AAS is also investigated.

Macromolecularly imprinted calcium phosphate/polyacrylate/alginate (CP/PA/A) multi-hybrid polymer microspheres with the surface and embedded imprinting of bovine serum albumin were prepared with sodium alginate (SA), (NH4)2HPO4 and sodium polyacrylate (SPA) by using CaCl2 as gelling agent in inverse-phase suspension. Infrared spectrum analysis demonstrated that multihybrid components were produced in the microspheres. Rebinding dynamic of both imprinted microspheres proved a higher BSA rebinding relative to those nonimprinted ones. The specific surface area of CP/PA/A microspheres increased when compared with non-imprinted ones. The specific rebinding peak and imprinting efficiency cross point have appeared in rebinding dynamic curves, which showed remarkable difference in rebinding behaviours between surface and embedding imprinted microspheres.

To eliminate the influence of serum on self-renewal of embryonic stem cells (ESCs), knockout serum replacement (KSR), a defined formulation, was used to replace serum for the establishment of C57BL/6J mouse ESC line.C57BL/6J mouse blastocysts collected at 3.5 days post coitum (d.p.c.) were cultured in the medium supplemented with KSR. In control experiment, KSR was substituted by fetal bovine serum (FBS). When ESC line was established, the morphology of ESCs, the expression of alkaline phosphatase and oct-4, and the karyotype and differentiating ability of ESCs were analyzed.13 blastocysts were cultured in the medium supplemented with KSR and one ESC line (MES-1) was established with a normal and stable XX karyotype after cultured for more than 20 passages, and then the high expression of alkaline phosphatase and oct-4 was detected. When cultured in suspension, MES-1 formed embryoid bodies. When inoculated subcutaneously into nude mice, MES-1 formed teratoma. After injected into ICR mouse blastocysts collected at 3.5 d.p.c., MES-1 incorporated into the inner cell mass of the host blastocyst and contributed to the development of a chimera. In control experiment, no ESC lines were cultured for more than 3 passages.KSR can be efficiently used to isolate and culture C57BL/6J mouse ESCs, which can eliminate traditional prescreening of FBS suitable for isolation and culture of ESCs.

The study of mammary gland bioreactor is in the ascendant. In order to generate transgenic goats of well-controlled expression of exogenic genes, we constructed a human lactoferrin (hLF) gene targeting vector containing promoter, exon 1, intron1 and some of exon 2 (about 6.1 kb fragment) and exon 6 approximately 9 (about 3.3 kb fragment) of the goat beta-casein gene as well as hLF minigene, neo gene inserted into them and tk gene ligated to the 3' end of the construct. The 9.4 kb goat genomic sequences as homologous arms were initially amplified by PCR with local goat tissue DNA. The expression vector was named pBC-tk-neo-hlf. Then the recombinant plasmid pBC-tk-neo-hlf containing hLF minigene was transfected into mice mammary tumor cell line C127 by liposome, cell clones were selected with G418. After proliferating, the transfected cells were induced with insulin, luteotropic hormone and hydrocortisone. The result of Western-blotting analysis showed that the transfected cells can secrete hLF protein, and the recombinant protein expressed in cultured cell supernatant has the similar molecular weight as the native protein. The expression level detected by ELISA was 0.21 microg/mL. This result indicated that the targeting vector could efficiently direct the expression of hLF in mammary cells,and it confirmed the validity of the constructed vector. At the same time, C127 cell line proved to be useful for evaluating the regulation of a foreign gene expression in mammary gland specific expression vector.

In order to improve the development rate of preimplantation nuclear transfer embryos (NT embryos) after transplanting nuclei derived from transgenic goat fetal cells, the donor fetal fibroblasts starved for 5 days in DMEM containing 0.5% FCS were divided into three groups and treated with different methods respectively before using as donor cell. Group 1 was frozen at -80 degrees C or in liquid nitrogen for several days or months. Group 2 was at first treated as the same as group 1, then cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. Group 3 was cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. The rate of G0/G1 phase cells from group 2 was 95.68% and significantly different from group 1's 88.66%. The rate of survival cells from group 2 was 99.9% and significantly different from group 1's 80.00% (P0.05).The morula- blastocyst stage NT embryos development rate of group 2 was 66.09% and significantly different from group 1's 22.00% and group 3's 50.51% (P0.05). All NT embryos of above three groups were transferred into synchronous oestrus recipients and the pregnant status of recipients was checked by B-mode ultrasound diagnosis after 35 days. The recipient pregnancy rate of group 2 was 45.83%, much higher than that of group 1(20.00%) and group 3 (29.58%). The result of this experiment showed that donor cells treated with freezing and two times starvation could significantly improve the rate of G0/G1 phase cells, the rate of survival cells, the NT embryos development rate and the recipient pregnancy rate.

To establish an animal model of HCV transgenic mice to elucidate the pathogenesis of hepatitis C virus infection and function of the viral structural proteins.Structural gene of HCV were amplified and recombined into eukaryotic expression vectors, pcDNA4HisMax and pMT/BiP/V5-His A, after their expressive activity was confirmed to detect the structural protein in the transfected COS7 and S2 cells by Western blot. The fertilized expression element, which contained CMV or pMT promoter, structural gene of HCV and polyadenylation signal sequence, was microinjected into 1736 C57BL/6 mouse fertilized ova. The ova were then replanted into the oviducts of 69 pseudopregnant recipient mice.Twenty-five recipient mice were impregnated and later produced 105 newborns; 49 of them died from unknown causes and 57 survived. After the specific HCV structural genes were identified by PCR and Southern blot hybridization, 26 founders were obtained; among them 10 were stable expression mice and 16 were the inducible ones. The rate of founders developed from implanted embryos was only 1.50%. Through hybridization with normal mice, 58 hybrid mice have been obtained at present.Two kinds of different transgenic mice of HCV were developed; one is of stable expression, and the other is inducible. This transgenic mice model may create an opportunity for studying the function of the structural gene of HCV and elucidate its pathogenicity.

To prepare monoclonal antibody (mAb) against erythropoietin(EPO), characterize its biological properties and use it to purify the rhEPO from transgenic goat milk.A crude rhEPO product was used as the antigen to immunize BALB/c mice for preparing mAbs against rhEPO. The mAbs were characterized by Western blot and indirect ELISA. Purified mAb 2E6 was coupled with pre-activated Sepharose 4B to prepare the immunoaffinity chromatography column for purifying the rhEPO from transgenic goat milk.Two hybridoma cell lines(1E7 and 2E6)were obtained. mAbs 1E7 and 2E6 were shown to be IgG1 and IgG2b respectively, and their light chains were both kappa. Western blot analysis confirmed that the two mAbs could bind to rhEPO. The immunoaffinity chromatography column could adsorb 70% of rhEPO in purifying the rhEPO from transgenic goat milk.Two hybridoma cell lines secreting anti-rhEPO mAbs were successfully established. The mAb-immunoaffinity chromatography column could be used to purify the rhEPO from transgenic goat milk.

Shaneng goat is a famous milking species. Boer goat is world famous goat breed for creophagism. In this study, we evaluated the development potential of adult Boer goat's somatic nuclei after nuclear tansfer (NT) into enucleated MII oocytes of the Shaneng goat. Somatic donor cells were obtained from two different sources: 1) adult granulosa cells (GCs) and 2) adult skin fibroblasts (FCs). The reconstructed embryos that developed to morula or blastocyst stage in vivo were transferred to 38 synchronized recipient.Somatic donor fibroblast cells were obtained from a fetal at 35 day. In the same way the reconstructed embryos were directly transferred to synchronized recipient of Shaneng goats. (1) Experimental group: NT embryos derived from GC and FC developed into morulas and blastocysts at a frequency of 46.8% and 31.4% respectively. Fifty-two NT morula and blastocyst stage embryos were transferred in to 38 recipients, Three of which were confirmed to be pregnant (7.9%). All pregnancies were not maintained to term. (2) CONTROL group: 136 NT embryos were transferred in to 14 recipients, Six of which were confirmed to be pregnant (42.9%). Four of those were maintained to term. Four recipients delivered four male kids (2.9% of embryos transferred). One male kid died at birth, the dead lamb shows as "large offspring syndrome". the others appeared health and normal. DNA analysis confirmed that those kids were genetically identical to their donor. These results demonstrated that Shaneng goat somatic cells could direct normal development and Shaneng goat oocyte cytoplasm supported development of preimplantation embryos produced by NT of somatic cell nuclei from Boer goat.

Nanocavity biomaterials with recognition specificity imprinted by using proteins as templates may successful serve as substitutes for antibodies, enzymes, and other native biological structures as well as cell bracket materials. It has numerous applications in biotechnology, medicine and so on. In this paper, the principle of template imprinting is introduced briefly, the specialty of template imprinting of proteins is analyzed, and the methods of template imprinting of proteins including protein entrapment, microbead surface imprinting, flat surface imprinting as well as the epitope are reviewed in details.

The cryopreservation of different embryo stages collected from ICR, C57BL/6 and F1 of DBA*C57BL/6 was carried out by using vitrification method. The morphology, in vitro development and birth rates of these embryos were compared after frozen-thawed. The results showed that more than 75% of the morphology from 2-cell embryos to morula stages from different strains was normal, the normal morphology rates of 8-cell embryos being the highest, while those of blastulas being the lowest. The in vitro development rates became higher as the embryos developed. The morphology of in vivo and in vitro fertilized frozen 2-cell embryos showed no difference, but the development rate of in vivo fertilized frozen 2-cell embryos was significantly higher than that of in vitro ones. Embryos that underwent 3 times frozen-thawing remained normal morphology. The pregnant rate and birth rate of frozen 2-cell embryos after embryo transfer were 64% and 40% respectively, but lower than those of fresh 2-cell embryo transfer.

To establish transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase specifically in the liver and to provide an efficient animal model for studying gene function in the liver and creating various mouse models mimicking human diseases.Alb-Cre-ERt transgenic mice were produced by microinjecting the construct with Cre-ERt fusion gene of DNA fragments into fertilized eggs derived from inbred C57BL/6 strain. Transgenic mice were identified by using PCR and Southern blotting. Expression of Cre-ERt fusion gene was analyzed in the liver, kidney, brain and lung from F1 generation transgenic mice at 8 weeks of age by reverse transcription (RT)-PCR.Four hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant Alb-Cre-ERt DNA fragments, and 312 survival eggs injected were transferred to the oviducts of 12 pseudopregnant recipient mice, 6 of 12 recipient mice became pregnant and gave birth to 44 offsprings. Of the 44 offsprings, two males and one female carried the hybrid Cre-ERt fusion gene. Three mice were determined as founders, and were back crossed to set up F1 generations with other inbred C57BL/6 mice. Transmission of Cre-ERt fusion gene in F1 offspring followed Mendelian rules. The expression of Cre-ERt mRNA was detected only in the liver of F1 offspring from two of three founder mice.Transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase under control of the liver-specific promoter are preliminary established.

Micro- and nano-structured surfaces of scaffold materials have important effects on cells' adhesion and proliferation. Nano-structured surfaces can improve cells' adhesion and biocompatibility of materials. The effects of nano-biomaterials on the development of tissue engineering and the methods of preparation of nano-biomaterials such as molecular self-assemble and template technology are discussed.

By combining liver-specific promoter and a chimeric Cre recombinase, conditional gene activation could be finely achieved in hepatocytes at selected time points. To this end, the expression vector of Cre-ERt under the control of the mouse albumin gene promoter/enhancer, alb-Cre-ERt, was constructed, and transfected into engineering BRL (Rat hepatocytes) and BRK (Rat kidney) reporter cells which carries a chromosomally integrated 'floxed' beta geo gene, which is inserted between the promoter and the human alkaline phosphatase( hAP) reporter gene, thereby preventing hAP reporter gene transcription, respectively. After treatment with 1 micromol/L 4-hydroxytamoxifen(4-OHT), a proportion of hAP staining positive cells were detected by hAP staining. It was further confirmed that 'floxed' beta geo cassette was removed by Cre excision by using PCR analysis of cellular DNA. No background recombinase activity could be detected in the absence of 4-OHT. Moreover, no hAP-positive cells could be detected in BHK cells untreated or treated with 4-OHT. These data suggested that alb-Cre-ERt expression vector was constructed successfully, and 4-OHT could induce Cre-mediated recombination only in hepatocytes expressing Cre-ERt, thereby activating a stably integrated hAP reporter gene. This provides a further foundation for producing transgenic mice expressing such an 4-OHT inducible Cre recombinase specifically in mouse liver.

A series of molecularly imprinted polymeric microspheres(MIPMs) were prepared by seed swelling and suspension polymerization method in aqueous system using tyrosine as printing molecules, methacrylic acid as functional monomers and trimethylolpropane trimethacrylate (TRIM) as cross-linkers. The morphology including the size, size distribution, pore and pore distribution of the polymer beads was analyzed by scanning electron microscope(SEM). The major factors that influence these properties of the beads are discussed. The molecule selecting property of the MIPMs was detected through liquid chromatography. The results showed that uniform-sized spherical MIPMs had been prepared in aqueous system by seed swelling and suspension polymerization method, and its surface was distributed with pores between 1 micron to 10 microns. The average bead size was adjusted from about 50 microns to 400 microns by changing the stirring speed and the amount of seeds used during the preparation. The prepared MIPMs exhibited a considerable tendency to adsorb tyrosine, the printing molecules selectively, and the separation factor was up to 1.82 when phenylalanine was chosen as the competitive molecules.

The ability to deliver nonviral DNA directly into animals would provide major practical and economic benefits. To pursue this potential, as a preliminary approach, the expression in mammary epithelial cells with nonviral DNA ending HBsAg and hEPO was investigated by in vivo delivery directly into mammary gland Either the bovine asl-casein-HBsAg or the WAE-hEPO gene construct was introduced into mammary glands of dm by mammary artery injection. A single injection of 10(11) copies of DNA construct was implemented for a doe. Transient expression of the constructs in milk collected from milk letdown by induction up to 19 d after injection was measured with an hEPO ELISA kit from Boehringer Mannheim (Cat. No. 1 693 417) for rhEPO, and a HBsAg-ELISA kit (Abbott Lab. USA) for rHBsAg. The treated animals showed mean concentration of recombinant molecules at 16 microg/L for rHBsAg, and 2 mg/L for rhEPO in milk.

AIM: To establish transgenic mice lineage harboring hepatitis B virus X gene and to provide an efficient animal model for studying the exact role of the HBx gene in the process of hepatocarcinogenesis. METHODS: The HBx transgenic mice were produced by microinjecting the construct with X gene of HBV (subtype adr) DNA fragment into fertilzed eggs derived from inbred C57BL/6 strain; transgenic mice were identified by using Nested PCR; expression and phenotype of HBx gene were analyzed in liver from transgenic mic at the age of 8 weeks by RT-PCR, pathologic examination and periodic acid-schiff staining (PAS), respectively. RESULTS: Five hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant retroviral DNA fragment, and 368 survival eggs injected were transferred to the oviducts of 18 pseudopregnant recipient mice, 8 of them became pregnant and gave birth to 20 F1 offspring. Of 20 offsprings, four males and two females carried the hybrid gene (HBx gene). Four male mice were determined as founder, named X1, X5, X9 and X15. These founders were back crossed to set up F1 generations with other ibred C57BL/6 mice or transgenic littermates, respectively. Transmission of HBx gene in F1 offspring of X1, X5 and X9 except in X15 followed Mendelian rules. The expression of HBx mRNA was detected in liver of F1 offspring from the founder mice (X1 and X9), which showed vacuolation lesion and glycogen positive foci. CONCLUSION: Transgenic mice harboring HBx gene were preliminarily established.

An electrochemical experimental method of cathodic blistering of coated metal under a galvanostatic charge condition has been proposed. By this means, corrosion resistance of three kinds of latexes, poly (methyl methacrylate) / poly (ethyl acrylate), PMMA/PEA (I);PMMA/PEA-polysiloxane (II); and PMMA/PEA-polysiloxane/ silica sol(III)in a 3% NaCl solution were evaluated and concentrations of sodium and chloride ions within blisters were analyzed. A cathodic blistering mechanism of coated galvanized steel panel is developed.