Objective To investigate the correlations of the single nucleotide polymorphism (SNP) 344-35C/T of growth arrest-specificgene 6 (GAS6) gene and cerebral infarction in Han population in the western Guangdong province. Methods The restriction fragment lengthpolymorphism (PCR-RFLP) was used to determine the 344-35C/T polymorphism in the intron region of GAS6 gene in the case group (n=180) and healthy control group (n=150). Results There was no significant association between GAS6-344-35C/T and cerebral infarction (P<0.05). After stratified by gender in chi-square test in women population, the frequency of C allele was significantly higher in cases (77.9%)than that in controls (66.4%) (P=0.040); The frequency of the CC genotype was significantly higher in cases (61.8%) than that in controls(43.1%) (P=0.036). Conclusion In Han women in western Guangdong province, C allele of GAS6-344-35C/T polymorphism is a risk factorof cerebral infarction, CC genotype is a susceptible genotype of cerebral infarction.

Background Epithelial-mesenchymal transition (EMT) is one of the pathogenesis mechanisms of posterior capule opcification (PCO).Studying EMT is of important significance for the prevention and treatment of PCO.However,EMT model is lack.Objective This study was to establish an in vitro EMT model for the study of human PCO.Methods In vitro mimic cataract enucleation was performed on 40 donor eyes,including anterior capsulorhexis,nucleus hydroexpression,and aspiration of lens fibers.The capsular bag of lens was dissected free during the surgery and pinned flat on a plastic culture dish with DMEM/F12 supplemented containing 10％ fetal bovine serum for 4 weeks.The proliferation of LECs on the capsular bag was observed by phase-contrast and dark-field microscope in 0,3,7,14 and 28 days after culture.The capsular bag tissues were collected in cultured 0,3,7 and 28 days for the preparation of sections and hematoxylin-eosin stain,and the growth and morphology of LECs were examined with optical microscope.The expression and location of α-SMA,E-cadherin and Vimentin were assessed by immunochemistry.The expression levels of α-SMA,E-cadherin and Vimentin mRNA and proteins were detected by using real-time fluorescnce quantitative PCR and Western blot in different time points.Results No LECs were seen on the uncultured capsular bag.LECs appeared in cultured day 3 on the periphery capsular bag and grew toward the center and covered the posterior capsule 7 days after cultured,with a cobblestone-like appearance.Wrinkles occurred and extended gradually along with the enhancement of bag tension.Immunochestry showed that the expression intensity of E-cadherin in the capsular bag gradually weakened,and that of α-SMA or Vimentin was gradually enhanced during the culture duration.The relative expression levels of E-cadherhin mRNA at 0,3,7,14 and 28 days after culture were 3.35±0.13,1.47±0.20,1.13±0.14,1.00±0.85 and 0.23±0.03,and relative expression levels of Vimentin mRNA were 1.00 ± 0.73,1.05 ± 0.14,2.24 ± 0.43,2.84 ± 0.34 and 8.57 ± 0.40,and those of α-SMA mRNA were 1.00 ± 0.06,2.68 ± 0.28,4.24 ± 0.05,2.05 ± 0.90 and 15.30 ± 0.19,showing significant differences among different time points (E-cadherhin mRNA:F =23.430,P =0.000;Vimentin mRNA F =8.915,P =0.002;α-SMA mRNA:F =103.500,P =0.000),with the lowest expression levels in the E-cadherhin mRNA and the highest expression levels in the Vimentin mRNA and α-SMA mRNA at 28 days during the culture period (all at P＜0.01).The gray values of E-cadherin protein expression were 1.443 ± 0.017,1.023 ± 0.003 and 0.568 ± 0.018,and those of Vimentin protein were 0.565±0.012,1.156±0.007 and 1.241±0.009,and those of α-SMA protein were 0.195±0.045,0.693±0.036 and 1.501±0.005 at 0,3 and 28 days,with significnant differences among various time points (E-cadherin:F =2 787.000,P =0.000;Vimentin:F =4 488.000,P =0.000;α-SMA:F =1 173.000,P =0.000).The expression levels were significantly declined in E-cadherhin protein and elevated in Vimentin and α-SMA proteins at 3 and 28 days after culture in comparison with before culture.Conclusions A novel in vitro EMT model of LECs is established in this study.This model can mimic a natural EMT procedure after extracapsular cataract enucleation and therefore is a useful model for the further research of the mechanism and prevention and treatment of PCO.

Objective To evaluate the association between the single-nucleotide polymorphism (SNP) of the 5-59A/G (rs2241027) of Htra2 gene and Parkinson's disease in Han population of the western GuangDong province. MethodsThe restriction fragment length polymorphism (PCR-RFLP) was used to determine the 5-59A/G polymorphism in the intron region of Htra2 gene in the case group (n=56) and healthy control group (n=109). ResultsA allele frequency of 5-59 A/G in cases (46.4%) was trended to more than that in controls (36.7%) (P=0.073), as well as the AA genotyping frequency (21.4% vs 11.0%, P=0.072). For the male, the frequency of AA genotype was significantly more in cases (25.7%) than that in controls (10.3%） (P=0.041), and the frequency of A allele was trended to more in cases (48.6%) than in controls (34.6%) (P=0.051). ConclusionA allele and AA genotype of the 5-59A/G (rs2241027) of Htra2 gene may increase the risk of suffering from Parkinson's disease, especially for males.

Objective To explore the association of ubiquitin-specific proteases 24 (USP24) gene polymorphisms with susceptibility to sporadic Parkinson's disease (PD) in the Han Guangdong population. Methods From August, 2006 to January, 2014, single nucleotide poly-morphisms (SNPs) of rs12138592 and rs6671533 in the intron region of USP24 were genotyped in 200 patients with sporadic PD and 200 healthy controls using the SNaPshot technique. Results There was significant difference in the allele and genotype frequency of rs12138592 between the patients and the controls (P<0.01), and no significant difference was found in the allele and genotype frequency of rs6671533 (P>0.05). Conclusion The SNP of rs12138592 in the intron region of USP24 is associated with the susceptibility to sporadic PD in the Han Guangdong population, and the A allele may contribute a protective roles to PD.

Objective To investigate the correlation between G753A and C1040T polymorphisms in the gene encoding region of thrombin-activatable fibrinolysis inhibitor (TAFI )and cerebral infarction in patients with cerebral infarction in Chinese Han population. Methods C1040T and G753A poly-morphisms in the TAFI gene encoding region in 130 patients with cerebral infarction and 118 healthy subjects (control group)were analyzed retrospectively and they were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP ). Results The GG genotyping of the TAFI gene G753A polymorphism in the cerebral group was 41. 5%(n=54)and the A allele carriers were 58. 5%(n=76),while those in the control group were 44. 9%(n=53)and 55. 1%(n=65)respectively. There were no significant differences in the GG genotyping of TAFI gene G753A polymorphism and the A allele carriers between the cerebral infarction group and the control group (χ2 =0. 288,P=0. 592). In the cerebral infarction group,the CC genotyping of C1040T polymorphism was 50. 0%(n=65)and T allele carriers were 50. 0%(n=65),while those in the control group were 51. 7%(n=61)and 48. 3%(n=57)respectively. There were no significant differences in the GG genotyping of C1040T polymorphism and the T allele carriers between the two groups (χ2 =0.071,P =0.790 ). Multivariate logistic regression analysis showed that G753A and C1040T single nucleotide polymorphisms (GA or AA genotype)in the TAFI gene encoding region were not the independent risk factors for cerebral infarction. Conclusion There are no significant differences in the correlation between the G753A and C1040T polymorphisms in the TAFI gene encoding region and cerebral infarction. They are not the independent risk factors for the onset of cerebral infarction.

ObjectiveTo investigate the relationship between Parkinson disease (PD) and intron rs12138592 A/G polymorphism of ubiquitin specific proteases (USP24) gene in Han population of the Western Guangdong province in China. Methods81 PD cases and 100 ethnically matched controls were investigated USP24 gene rs12138592 A/G polymorphism with polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). ResultsThe incidence of GG genotype was 77.8% in the cases and 62.0% in the controls (χ2=5.213,P=0.022), and the G allele was 88.3% in the cases, 79.5% in the controls (χ2=4.980，P=0.026). ConclusionThe G allele and GG genotype of USP24 gene rs12138592 A/G polymorphism can increase the risk of suffering from PD.

Abstract: Objective To research the relationship between promoter polymorphism-438 A/G of thrombin-activatable fibrinolysis inhibitorgene (TAFI-438 A/G) and atherosclerotic cerebral infarction (ACI) in Chinese Han population. Methods TAFI-438A/G genotypes andtheir allele frequencies were identified with the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in225 ACI patients and 184 healthy controls. Results There was no significant difference of TAFI-438A/G polymorphism between ACI groupand control group. Stratified by gender, in males, the incidence of A allele was 28.6% in ACI group, and 20.6% in control (P=0.039); of theAA genotype was 9.0% in ACI group, and 1.9% in control (P=0.019). But no significant difference was found in females. ConclusionTAFI-438A/G polymorphism is associated with the risk of ACI in males, that AA genotype may increase the risk of ACI.

Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P

Objective To study the effect of the uncellular tissue engineering complexes of autolegous red bone marrow wrapped by facial flap with vessels in repair of large segment bone defect infected with low virulence bacteria so as to provide evidence for the clinical application. Methods The study included 38 cases of limb bone defect infected with low virulence bacteria after trauma.Autologous red bone marrow (ARBM) was taken to prepare uncelluar tissue-engineered complexes with osteoinductive absorbing material (OAM) containing bone morphogenetic protein (BMP).A facial flap with capillary network originating from an anonymous vessel adjacent to the bone defect was prepared to wrap the tissue engineered bone and fill the bone defect.Pathological focus clearance and tissue-engineered complexes compounded with ARBM implantation were performed in 18 cases (Group A) and pathological focus clearance and tissue-engineered complexes of autolegous red bone marrow wrapped by facial flap with vessels implantation in the other 20 cases ( Group B).The blood routine and supersensitive CRP were examined to monitor the inflammation reaction; X-ray was used to observe the bone defect repair; histology and bacteriology examinations were performed in partial cases at 3,6,12,18 months after operation. Results Six months after operation,5 cases of Group A were infected and the bacteria cultivation was as positive as that before the operation.The histological observation at ( 14.0 ± 0.5 ) months after operation showed that fibrous connective tissues between the bone fracture ends existed in the pathological area in 10 cases,of whom four cases were filled with inflammatory fibrous granulation tissues and few dead bones in the pathological area,and the bacterial examination was positive.There was no infection in Group B after operation.The histological observation manifested periosteum like tissues formation from the primary facial flap,mature bone structure formation in the primary pathological area and non-inflammatory infiltration in 16 cases and the bacteria cultivation was negative in these cases.The external fixation frame was taken out (12.2 ± 0.3 )months after operation because the synostosis appeared and the structure was stable in the other seven cases including three cases in Group A and four in Group B and the histological and bacterial examination were not performed.At each time point after operation,not only the blood routine but also the supersensitive CRP and the X-ray quantification grade of Group B were significantly more than those of Group A (P ＜ 0.05 ). Conclusions The uncellular tissue-engineered complexes of autolegous red bone marrow wrapped by facial flap with vessels is a feasible method for repairing the infected bone defect by first intention,since it can resist infection,obviously promote the bone recovery and advance the quality and quantity of osteanagenesis.

A total of 38 cases of senile brucella spondylitis disease at our hospital during January 2002 and March 2012 were analyzed .After admission , all of them were definitely diagnosed on the basis of epidemiological history , clinical manifestations , laboratory tests , imaging and pathological examinations . Over a follow-up period of 12 months, 17 cases were cured after standardized drug treatment .Among 21 surgical cases, there were curing (n=17) and improving (n=2).Senile brucellosis spondylitis has distinct serological and pathological characteristics .And formulating the diagnostic criteria may improve its diagnostic rate and reduce its misdiagnostic rate .And standardized drug therapy achieves a better curing rate and a proper timing of surgical intervention improves its clinical outcomes .