1.Correlation between late-onset sporadic Parkinson's disease and single nucleotide polymorphism of Atg7 rs2606757
Xiyao ZHAO ; Yusen CHEN ; Li WANG ; Xiangxin LI ; Xiaoyi CHEN
Chinese Journal of Geriatrics 2019;38(1):10-13
Objective To investigate the correlation between late-onset sporadic parkinson's disease(PD) and single nucleotide polymorphism (SNP) of autophagy-related genes 7 (Atg7) rs2606757 (6+1196A/T)in Han Chinese population.Methods Totally 124 patients with late-onset sporadic PD(the PD group)and 105 age-and sex-matched healthy individuals(the control group)were enrolled in this study.The SNP of Atg7 rs2606757 was detected by the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).After gene sequencing for the detection of allele and genotype frequencies distribution and testing Hardy-Weinberg equilibrium,the differences in Atg7 rs2606757 genotype and allele frequency were compared between PD group and control group and between males and females.Results The frequency of AA genotype was statistically significantly lower in the PD group[34.7%(43/124)]than in the control group[53.3 % (56/105)](x2 =8.063,P=0.005,OR =0.465,95% CI:0.273-0.791).In men's PD group versus men's control group,AA genotype of the Atg7 rs2606757 showed a lower frequency for late-onset sporadic PD[33.3 % (23/69)vs.53.2 % (33/62),x2 =5.280,P =0.022,OR =0.439,95 % CI:0.217-0.891].Logistic regression analysis indicated that the AA genotype frequency distribution of Atg7 rs2606757 showed a significant difference between PD and control groups (OR =2.210,95% CI:1.289-3.789,P =0.004).Conclusions The higher frequence of AA genotype at Atg7 rs2606757 only in males might be associated with the decreased risk of late-onset sporadic PD.
2.Association between Ubiquitin-specific Proteases 24 Gene Polymorphisms and Sporadic Parkinson's Disease in the Han Guangdong Population
Zhijun LIN ; Yusen CHEN ; Wangtao ZHONG ; Zhou LIU ; Wenchuan XIAN ; Xiaoyi CHEN ; Bin ZHAO
Chinese Journal of Rehabilitation Theory and Practice 2017;23(3):345-348
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.
3.The analysis of correlation between single nucleotide polymorphism at Atg7 rs14016 and Parkinson's disease
Xiyao ZHAO ; Yusen CHEN ; Xiaoyi CHEN ; Li WANG ; Xiangxin LI
Chinese Journal of Geriatrics 2017;36(12):1293-1297
Objective To investigate the association between rs14016 (19 +31C/T) polymorphisms of autophagy-related genes 7 (Atg7) and Parkinson's disease (PD) in Han population in China.Methods Totally 123 patients with Parkinson's disease (PD) (case group) and 101 synchronized health controls (control group) were selected from Chinese Han population between January 2013 and July 2016.A single nucleotide polymorphism (SNP) of rs14016 of Atg7 gene was detected by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis.After gene sequencing for genotyping and detection of alleles,genotype and allele frequency distributions were analyzed in the two groups.Results The frequency distributions of TT genotype and T alleles were 17.9% (22/123) and 41.1% (101/246) in the case group,and 5.9% (6/101) and 31.2 % (63/202) in the control group,respectively.The difference in genotype frequencies between the two groups was statistically significant (x2 =7.236,P =0.007,OR =3.01,95 % CI:1.27-7.14).The frequencies of T and C alleles were 41.1% (101/246) and 58.9% (145/246) respectively in the case group,and 31.2% (63/202) and 68.8% (139/202) in the control group (x2 =4.655,P=0.031,OR=1.32,95 %CI:1.02-1.70),with the statistically significant difference in the allele.The TT genotype of rs14016 showed statistical significance between the two groups by logistic regression analysis (OR=3.40,95%CI:1.32-8.80,P=0.012).Conclusions The T allele and TT genotype at the rs14016 of Atg7 gene might be associated with PD,and might increase the risk for suffering from PD,which is worthy of further fully researches.
4.Effect of targeted monitoring and comprehensive intervention on reducing catheter-associated urinary tract infection in patients in non-intensive care unit
Li KONG ; Yusen LI ; Zhankui ZHAO ; Huifang HU ; Jinfeng GUO ; Hong LIU ; Juan WANG ; Jixia WANG ; Miao LIU ; Ning LI ; Xia YUE ; Dingding ZHANG ; Yonghui MA
Chinese Journal of Infection Control 2017;16(8):721-725
Objective To evaluate the effect of targeted monitoring and comprehensive intervention measures on reducing the occurrence of catheter-associated urinary tract infection(CAUTI)in patients in non-intensive care unit(Non-ICU).Methods In quarter 4 of 2015,patients with indwelling urinary catheter in clinical departments were conducted a baseline survey(before intervention),risk factors for CAUTI in patients were analyzed,targeted monitoring programmes and comprehensive intervention measures were initiated in 2016(after intervention),incidence of CAUTI before and after intervention was compared.Results After taking intervention measures,hand hygiene compliance rate increased from 78.51%in quarter 4 of 2015 to 92.99%in quarter 3 of 2016 and 90.73%in quarter 4 of 2016(x2=7.342,3.998,respectively,both P<0.05),the correct disposal rate of patients' urinary catheterization system increased from 72.83%in quarter 4 of 2015 to 95.44%in quarter 4 of 2016(x2=30.267,P<0.05).A total of 12 067 patients with indwelling urinary catheter were monitored,incidence of CAUTI dropped from 1.03%(24/23 313)in quarter 4 of 2015(before intervention)to 0.53%(14/26 595)in quarter 4 of 2016(after intervention),difference was statistically significant(x2=4.126,P=0.042).Conclusion Improving the quality of urinary catheterization system in patients with indwelling catheter through targeted monitoring can effectively reduce the incidence of CAUTI in patients in Non-ICU.
5.In vitro epithelial-mesenchymal transition model for LECs of human posterior capsule opacification
Baoxia, YANG ; Ye, WANG ; Xiaowen, ZHAO ; Ting, LIU ; Yusen, HUANG
Chinese Journal of Experimental Ophthalmology 2016;34(3):210-217
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.
6.Comparison of pulmonary pathological changes in mice infected with H7N9 influenza virus and pandemic H1N1 influenza virus
Shihui SUN ; Xiaohong WU ; Chenfeng LIU ; Tongtong GAO ; Yang ZENG ; Yan GUO ; Jian TANG ; Ting PAN ; Hong YU ; Zhihua KOU ; Guangyu ZHAO ; Yusen ZHOU
Acta Laboratorium Animalis Scientia Sinica 2014;(3):1-6
Objective To analyze and compare the pathological changes of lung tissue in mice infected with the novel H7N9 influenza virus and 2009 pandemic H1N1 influenza virus, respectively, and to preliminarily study the mecha-nisms of acute lung injury induced by those virus infection .Methods SPF 6-week old BALB/c mice ( body weight 18-20 g, male∶female=1∶1) (n=3 in each subgroup) were intranasally infected with H7N9 virus and H1N1 virus, respec-tively.The behavior and survival time of mice after virus infection were observed and the survival rates were analyzed .The heart, liver, spleen, lung, kidney, intestines, and brain were collected at indicated time points for histopathological exami-nation using H&E staining .The distribution of virus antigen was detected by immunohistochemistry .The neutrophil infiltra-tion was also observed .The correlation of lung injury with virus replication and host immune responses was analyzed .Re-sults The lung and spleen injury of mice infected with H 7N9 virus was slighter and their survival rate (100%) was high-er than those of mice infected with H1N1 virus.The damages of the lung and spleen in H1N1virus-infected mice were more severe than that in H7N9 virus-infected mice, and all the 10 mice in this group died within 9 days after virus inoculation . The distributions of both the virus antigens were mainly in the bronchial epithelial cells , a few stromal cells and alveolar ep-ithelial cells .The levels of virus replication in the two groups were not significantly different .There were more intense neu-trophil infiltration in the lung and inflammatory response in the H 1N1 virus-infected mice than those in the H7N9 virus-in-fected mice .Conclusions There are some differences of the pathological characteristics and extent of lung injury in the mice infected with H7N9 virus and H1N1 virus, respectively.The virus replication is a precipitating factor but not the deci-sive factor of the lung injury , and there is a close relationship between the host immune responses and acute lung injury .
7.Correlation between C1040T and G753A polymorphisms in the gene encoding region of thrombin-activatable fibrinolysis inhibitor and cerebral infarction
Fangmei HE ; Jiangang PAN ; Xiyao ZHAO ; Hua YUAN ; Xiang MOU ; Yusen CHEN
Chinese Journal of Cerebrovascular Diseases 2014;(7):347-353
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.
8.Diagnosis and treatment of senile brucellosis spondylitis
Xinming YANG ; Wei SHI ; Xianyong MENG ; Changbo HU ; Peng ZHANG ; Yaoyi WANG ; Yongli JIA ; Zhenshun HU ; Yusen ZHAO
Chinese Journal of General Practitioners 2014;(5):386-387,388
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 .
9.Repair of low virulence bacteria limb bone defect with uncellular tissue-engineered complexes of autolegous red bone marrow wrapped by facial flap with vessels
Xinming YANG ; Lei ZHANG ; Ying ZHANG ; Yaoyi WANG ; Xianyong MENG ; Zhenshun HU ; Junwei ZHANG ; Peinan ZHANG ; Yusen. ZHAO
Chinese Journal of Trauma 2012;28(1):54-60
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.
10.Construction of anti-H5N1 virus chimeric igA antibody gene and its expression in CHO cells.
Baozhong ZHANG ; Xin ZHANG ; Wanrong CHEN ; Dabin LIU ; Sheng WANG ; Xiaoping AN ; Duoliang RAN ; Guangyu ZHAO ; Yusen ZHOU ; Yigang TONG
Chinese Journal of Biotechnology 2009;25(5):714-719
Abstract: To express human-mouse chimeric IgA antibody directed against H5N1 virus, an anti-H5N1 chimeric IgA antibody gene was constructed by joining the light and heavy chain variable region genes and the corresponding signal peptide coding sequences of the anti-H5N1 mouse monoclonal antibody H5N1-HA with the coding sequences of the constant region of the human IgA2 heavy chain and Kappa chain respectively. Then the full-length chimeric light and heavy chain expressing plasmids pEF-IGHA9 and pEF-IGK9 were constructed and transfected into the CHO/dhfr cells. The chimeric IgA antibody expression was confirmed by ELISA, SDS-PAGE and Western blotting. The successful expression of this anti-H5N1 chimeric IgA may help to provide a stand for developing passive immunological agents for H5N1 virus infection prophylaxis.
Animals
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Antibodies, Monoclonal
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biosynthesis
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Antibodies, Viral
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genetics
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CHO Cells
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Chimerism
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Cricetinae
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Cricetulus
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Humans
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Immunoglobulin A
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genetics
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immunology
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Influenza A Virus, H5N1 Subtype
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genetics
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immunology
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Mice
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Recombinant Fusion Proteins
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biosynthesis
;
genetics
;
immunology

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