2.Transient expression and biological activity identification of human pigment epithelium-derived factor in mammary cell line SP2/0
Yi DAI ; Wenjing SHI ; Yuxiong WANG ; Min YU ; Chao CHEN
Chinese Journal of Perinatal Medicine 2011;14(6):347-353
Objective To construct eukaryotic expression plasmid pIRESneo3-pigment epithelium-derived factor (PEDF) and detect its transient expression in SP2/0 cells. Methods Specific primers were designed based on the mature peptide sequence of human PEDF cDNA in the GenBank. Human PEDF gene was cloned into the eukaryotic expression vector pIRESneo3. The PEDF DNA was transfected into SP2/0 with LipofectamineTM 2000. The recombinant human PEDF protein expressed in SP2/0 cell culture supernatant was identified by Western blot and enzyme-linked immunosorbent assay. The biological activity of the recombinant human PEDF was measured by 3-(4,5-dimethylthiazol-z-y1)-2,5-diphenytetrazolium bromide(MTT) method. Results PCR amplification, restriction enzyme digestion and DNA sequencing confirmed that the mature peptide sequence of human PEDF cDNA was successfully cloned into the eukaryotic expression vector pIRESneo3. And the plasmid was transfected into SP2/0 cells, which could secret PEDF. Western blot analysis showed that there was only one obvious band at the position of relative molecular weight of 50 000, and it is equivalent to the expected value. Enzyme-linked immunosorbent assay suggested that the content of PEDF began to rise after transfection, and peaked at 36 h [(0.92±0.04) μg/ml]. The proliferation of human umbilical vein endothelial cell line was significantly inhibited by supernatant after transfection of 36 h (P<0.05). Conclusions The eukaryotic expression plasmid pIRESneo3-PEDF had been successfully constructed and active human PEDF was transiently secreted, which made a foundation for further study of stable expression and purification of PEDF. This protein could be a potential medication for preventing and managing retinopathy of prematurity.
3.Ultrastructural observation of morphologically abnormal sperm: Advances in studies and application.
Jia-xiong WANG ; Yi-chao SHI ; Shen-min YANG
National Journal of Andrology 2016;22(1):67-71
Sperm ultrastructural abnormalities are often associated with sperm motility, the integrity of genetic material, and the fertilization potential. The investigation of sperm ultrastructural abnormalities is based on the evolution of microscopy techniques. In his paper, we review the improvement of the microscopy techniques and the ultrastructure of several specific morphological defects and he apoptotic spermatogenic cells in order to expound the significance of sperm ultrastructural observation in clinical practice. We deem it necessary to analyze the sperm ultrastructure before exploring the pathology and adopting assisted reproductive technology for some special patients with teratozoospermia.
Humans
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Male
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Microscopy
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Spermatozoa
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abnormalities
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ultrastructure
4.Value of soluble epithelial cadherin in differential diagnosis of pleural effusion
ling, ZHOU ; yi, XIANG ; guo-chao, SHI ; huan-ying, WAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
Objective To investigate the value of soluble epithelial cadherin(sE-cad)in the differential diagnosis of pleural effusion. Methods Patients were divided into malignant pleural effusion group,infective pleural effusion group and transudation group.sE-cad in pleural fluids obtained during the first thoracocentesis was measured by enzyme-linked immunosorbent assay(ELISA).The concentration of sE-cad in all kinds of pleural effusions was compared.The cut-off value of sE-cad for the differential diagnosis of benign and malignant pleural effusion was determined by ROC curve.The diagnostic value of sE-cad was also compared with common tumor markers such as CEA,CA199,CA125 and NSE.Results The concentration of sE-cad was significant higher in the malignant pleural effusion than in the benign pleural effusion[(38.38?4.15)ng/mL vs(14.17?0.80)ng/mL,P
5.Clinical analysis of 41 cases of Hashimoto's disease accompanying with thyroid cancer
Zhipeng ZHANG ; Yi XIA ; Huijun LIAO ; Chao DONG ; Shi CHANG
Journal of Endocrine Surgery 2013;7(4):281-283
Objective To analyze the clinical characteristics of Hashimoto's disease(HD) accompanying with thyroid cancer,and to explore the experience of diagnosis and treatment.Methods Clinical data of 41 cases of HD accompanying with thyroid cancer were retrospectively analyzed.The patients were diagnosed by postoperative paraffin pathological examination from Jan.2002 to July 2011.Results 10 cases of HD,37cases of thyroid cancer,and 8 cases HD accompanying with thyroid cancer were diagnosed before operation.The rate of preoperative diagnosis was only 19.51%.All patients underwent surgical treatment,including 22 cases total thyroidectomy,14 cases subtotal thyroidectomy,and 5 cases lesion side lobe resection.24 cases underwent ipsilateral neck dissection,and 4 cases underwent bilateral neck dissection (ipsilateral radical resection,contralateral selective resection).Postoperative paraffin pathological examination proved that there were 39 cases of HD accompanying with thyroid cancer,1 case of focal cancer and 1 case of B-cell lymphoma of mucosa-associated thyroid.All patients were followed up.Conclusions The preoperative diagnosis rate of HD accompanying with thyroid cancer is low and great attention should be paid to its diagnosis.For HD patients,if carcinoma can not be excluded,surgical exploration is recommended.Appropriate surgical method should be chosen according to intraoperative frozen section results.Postoperative thyroid hormone treatment is usually taken.
6.Influencing factors of standardization in the hierarchical chain management of type 2 diabetes
Yi WANG ; Qingge GAO ; Xiangying MENG ; Yanrong LU ; Chao SHI ; Rong CHEN ; Changxiu LIANG ; Yong ZHOU
Clinical Medicine of China 2012;28(12):1264-1268
Objective To analysis the influence factors of standardization in the hierarchical chain management of type 2 diabetes and to enhance the hierarchical chain management of type 2 diabetes.Methods ( 1 ) Six hundred and ninty patients with type 2 diabetes completed 1 years management were divided into well-controlled glycosylated hemoglobin ( HbAlc ) group (<7.0% ) and bad-controlled glycosylated hemoglobin (HbAlc) group ( ≥ 7.0% ).The conditions of diet,physical activity,medication,self-blood sugar monitoring and participation in health seminars were investigated and analyzed.(2) The patients were divided into standardized management group and not standardized management group.Their age,sex,educational background,occupation,monthly income per person,medical security,the course,cognition for glycuresis,two-way transfer,and chronic complications were investigated and statistically analyzed.Results ( 1 ) The proportions of physical activity (70.1% vs 54.2%,x2=6.163,P=0.018),self-blood sugar monitoring(60.4% vs 43.8%,x2=6.268,P=0.016) and participation in health seminars (56.0% vs 41.7%,x2=4.577,P=0.045) in the well-controlled HbAlc group were significantly higher than those in the bad-controlled HbAlc group.(2) Their age [(61.08 ±10.04) years old vs ( 57.75 ± 9.89 ) years old,t=2.539,P=0.012],educational background ( ratio of low educational attainment:8.3 % vs 17.2%,x2=6.426,P=0.041 ),medical security (own expense ratios:4.6% vs 11.5%,x2=3.543,P=0.048 ),awareness of diabetes ( ratio of poor awareness of diabetes:19.4% vs 41.0%,x2=17.518,P=0.000 ),two-way transfer ( ratio of not transfer treatment:4.6% vs 14.8%,x2=7.662,P=0.022) and chronic complications ( ratio of chronic complication:41.7 % vs 26.2%,x2=6.130,P=0.017) were significantly different between the standardized management group and not standardized management group.(3) Logistic regression analyses indicated that the age ( OR=0.954,P=0.006),monthly income per person ( OR=4.101,P=0.018 ),medical security ( OR=7.617,P=0.003 ),cognition for glycuresis ( OR=0.030,P=0.000),two-way transfer ( OR=9.079,P=0.000) and chronic complications ( OR=0.456,P=0.031 ) were the risk factors of standardized management.Conclusion We should focus on the impact factors affecting the standardized management of patients including age,monthly income per person,medical security,awareness of diabetes,ratio of not transfer treatment,positive strategies for chronic complications,improve the hierarchical chain management of type 2 diabetes,and then make the diabetic patients to early participate in standardization management of diabetes mellitus and delay the appearance of complications.
7.Study and effect assessment of the hierarchical chain management model of type 2 diabetes
Qingge GAO ; Yi WANG ; Chao SHI ; Rong CHEN ; Changxiu LIANG ; Yanrong LU ; Yong ZHOU
Chinese Journal of Postgraduates of Medicine 2011;34(16):28-31
Objective To explore the hierarchical chain management model of type 2 diabetes and determine its evaluation.Method Based on the hierarchical chain management of the three community health service institutions and Dahua hospital in Shanghai Xuhui district,215 cases of type 2 diabetes had been involved in the study.Results Compared with the baseline before management,lasting blood glucose (FBG),2 h postprandial glucose (2hPBG),glycosylated hemoglobin (HbA1c),low density lipoprotein cholesterol (LDL-C),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the diabetes after 12 months' management declined [(8.50 ±2.81) mmol/L,(11.09 ±4.01) mmol/L,(8.56 ±2.41)% ,(3.31 ± 1.06) mmol/L,(139.06 ±20.68) mm Hg (1 mm Hg = 0.133 kPa),(78.20 ± 12.11) mm Hg vs.(7.41 ±2.04) mmol/L,(9.03 ±2.46) mmol/L,(7.34 ± 1.59)% ,(3.00 ± 1.06) mmol/L,(135.48 ± 17.82) mm Hg,(77.27 ±11.83) mm Hg],and the differences were statistically significant(P<0.01 );control rate of FBG,2hPBG,HbA1c,LDLC,SBP,DBP had improved significantly [19.5% (42/215),20.9% (45/215),24.7%(53/215),20.0%(43/215),27.4%(59/215),30.2%(65/215) vs.50.7%(109/215),53.0% (114/215),54.0%(ll6/215),42.3%(91/215),47.0%(101/215),45.6%(98/215)](P<0.01).Conclusion Primary and secondary-care hospital based hierarchical chain management model is valid and can be implemented for type 2 diabetes.
8.Clinical analysis of bacterial infection in liver transplant recipients
Rui GAO ; Yi Lü ; Chang LIU ; Zhantao XIE ; Chao ZHAI ; Jianhua SHI ; Zhen WAN
Journal of Xi'an Jiaotong University(Medical Sciences) 2009;30(6):683-686
Objective To explore the incidence and risk factors of bacterial infection after othtotopic liver transplantation (OLT). Methods Altogether 56 OLT recipients from January 2005 to October 2007 were included in the study. The incidents and the related variables of the infection were analyzed retrospectively. The related variables were evaluated using multivariate logistic regression model to identify the significant risk factors. Results Bacterial infection was confirmed in 29 recipients (51.8%). Among them, the lung infection was the most common site (53.7%). The Gram-positive cocci were 46.3%, while the Gram-negative bacilli were 53.7%. The risk factors for bacterial infection included duration of the operation and detained respirator using. Conclusion Bacterial infection is a major complication following OLT. Surveillance for the risk factors, enhancement the skill of operation, and improving the recovery of respiratory function is the key to decreasing the incidence of bacterial infection after transplantation.
9.Effect of low molecular weight heparin on clot rate in vitro
Xubo SHI ; Dayi HU ; Hua ZHENG ; Hong BIAN ; Chao LI ; Yi YANG
Chinese Journal of Laboratory Medicine 2011;34(1):26-29
Objective To study the in vitro effects of different doses and different kinds of LMWH on CR, and to determine whether the CR test could be used to monitor LMWH. Methods The CR value was measured with different reagents ( glass beads, celite and kaolin ) in blood samples from twenty volunteer donors, which were spiked with increasing concentration of LMWH ( dalteparin, 0-1.8 IU/ml ). Then the CR test was performed again on the same blood samples spiked with the same concentration ( 0. 8 IU/ml ) but different LMWH ( dalteparin, enoxaparin and nadroparin ). Regression analysis was performed to establish a regression equation from corresponding LMWH levels. Results With the increasing of dalteparin dose, CR values were reduced gradually for all three reagents. When the concentration of dalteparin was 0-1.8 IU/ml,the value of CR was 20. 0-4. 5 IU/min for glass beads, 26. 1-6.6 IU/min for celite and 27. 2-7. 5 IU/min for kaolin. An exponential relationship was observed between the CR value and dalteparin concentration for three reagents( R2 = -0.796, -0.884, -0.921 ,P <0.01 ). All three kinds of LMWH with the same concentration (0.8 IU/ml ) induced a different change in CR. The value of CR was 7.4 IU/min with dalteparin,8. 5 IU/min with enoxaparin and 8.5 IU/min with nadroparin. Compared with the control group ( CR was 17.6 IU/min ), three kinds of LMWH had statistical significance ( t = 18.45, 12. 33, 14. 93, P < 0.01 ).Compared with the enoxaparin and nadroparin, dalteparin induced a higher CR value ( t = 2. 552,2. 924,P<0. 05 ). Conclusions There is an exponential relationship between CR value and dalteparin concentration for three reagents. Three kinds of LMWH can significantly reduce the value of CR. CR test can be used to monitor the anticoagulant effect of LMWH.
10.Long-term Prognostic Analysis of Re-operation in Patients With Functional Tricuspid Regurgitation After Left-sided Valve Replacement
Minghui TONG ; Yi SHI ; Shen LIU ; Xiang LUO ; Chao DONG ; Yan YANG ; Wei WANG ; Jianping XU
Chinese Circulation Journal 2016;31(4):376-380
Objective: To analyze the long-term prognosis of re-operation in patients with functional tricuspid regurgitation (FTR) after left sided valve replacement (LSVR) and hence evaluate the optimal timing of mentioned re-operation. Methods: A total of 59 FTR patients who had re-operation after their prior LSVR in our hospital from 1999-01 to 2013-01 were analyzed. The clinical information and post-operative follow-up results were recorded in all patients. Results: There were 5/59 (8.5%) patients died in peri-operative period and the overall post-operative mortality was 11.9% (7/59). The follow-up data of 54 survivors were available for the mean time of 51.1 (21-188) months. There were 19/54 (35.2%) patients suffered from MACE and 30 (55.6%) were beneifted by improved cardiac function. Uni-variable analysis indicated that pre-operative NYHA class IV (P=0.008), pre-operative right ventricular (RV) dysfunction (P=0.037), concomitant left-sided redo-operation (P=0.017) and TVR operation (P=0.002) were associated with all cause mortality of tricuspid re-operation. Multi-variable Cox regression analysis showed that pre-operative RV dysfunction was the only independent risk factor of long term MACE-free accumulating survival rate (HR=3.0, 95% CI 1.11-8.2,P=0.031); while TVR operation (HR=12.8, 95% CI 1.53-107.02,P=0.019) and pre-operative NYHA class IV (HR=5.3, 95% CI 1.20-24.51,P=0.032) were the independent risk factors for long-term mortality in patients after tricuspid re-operation. Conclusion: Patients with compensatory RV function showed better long term prognosis after secondary tricuspid operation. Aggressive re-operation before the occurrence of right ventricular dysfunction could be beneficial for relevant patients.