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.

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

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.

China ; Humans ; Occupational Health Services

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 ; Male ; Microscopy ; Spermatozoa ; abnormalities ; ultrastructure

Objective To explore the key signaling pathways and molecules of bone sialoprotein (BSP) to promote proliferation and differentiation of preosteoclasts RAW264.7. Methods RAW264.7 cells were treated with BSP and the inhibitors of signaling pathway molecules. MTS and TRAP staining kits were used to evaluate the effects of proliferation and differentiation. The activity assay kits of Calcineurin, AKT, JNK, ERK and p38 were used to detect their activities. Results Inhibiting ERK and p38 can inhibit cell proliferation induced by BSP significantly, while inhibiting AKT, Calcineurin and PI3K can reduce the role of promoting the differentiation by BSP. With increasing treatment time of BSP, the activity of Calcineurin in RAW264.7 cells increased. Furthermore, the activity of AKT was maximum at 48 h after treated with BSP, while the activity of JNK, ERK and p38 were maximum at 24 h after treated with BSP. Conclusions BSP mainly regulates the ERK and p38 of MAPK pathway to promote RAW264.7 cell proliferation, and regulates AKT, PI3K and Calcineurin pathways to promote RAW264.7 cell differentiation.

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.

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.

OBJECTIVE:To observe the efficacy and safety of edaravone combined with nimodipine in the treatment of acute large area cerebral infarction. METHODS:78 patients with acute large area cerebral infarction were randomly divided into control group (38 cases) and observation group (40 cases). Control group received aspirin platelet aggregation,reduced intracranial pres-sure,maintained water and electrolyte balance,decreased cerebral edema,Xuesetong for injection,Ginkgobiloba injection,lipid and blood pressurelowering,maintained blood sugar and other conventional treatment;observation group additionally received 30 mg Edaravone injection,adding into 100 ml 0.9% Sodium chloride injection by intravenous infusion (drip completed within 30 min),twice a day+12 mg Nimodipine injection,adding into 500 ml 0.9% Sodium chloride injection by intravenous infusion,once a day,with drip rate of 0.5-1 mg/h,for continuous 10 d,then received 40 mg Nimodipine tablet,orally,3 times a day. The treat-ment course for both groups was 15 d. Clinical efficacy,blood oxygen saturation (SpO2),hemoglobin (Hb),total hemoglobin (HbT),reduced hemoglobin(MHb)before and after treatment,and the incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than control group,the incidence of adverse reac-tions was significantly lower tahn control group,the differences were statistically significant (P<0.05). Before treatment,there were no significant differences in SpO2,Hb,HbT and MHb in 2 groups (P>0.05). After treatment,the SpO2,HbT and MHb were significantly higher than before,and observation group was higher than control group,the differences were statistically signifi-cant(P<0.05);Hb was significantly lower than before,the differences were statistically significant(P<0.05),but there was no significant difference between 2 groups (P>0.05). CONCLUSIONS:Based on the conventional treatment,edaravone combined with nimodipine shows significant efficacy in the treatment of acute large area cerebral infarction,it can effectively recovery neuro-logical function damage and protect the brain tissue,which can reduce the incidence of adverse.

Objective To explore the inhibitive effect of BSP on Caspase dependent apoptosis of preosteoclasts RAW264.7 cells by bone sialoprotein (BSP).Methods RAW264.7 cells were treated with the inhibitors of signaling pathway molecules and/or BSP.The activities of caspase3/8/9 in RAW264.7 cells were detected with caspase3/7/8/9 kits and the interaction between Caspase-8 and integrin αVβ3 was detected by Co-IP.Results The activities of Caspase3/8/9 in BSP-treated RAW264.7 cells were decreased and the apoptosis of RAW264.7 cells were inhibited significantly.In addition,there was a positive correlation between caspase-8 activity and caspase-3 activity.There was interaction between caspase-8 and integrin αVβ3.Conclusion The anti-apoptotic effect of BSP on RAW264.7 cells is mainly through the regulation of exogenous apoptosis pathway and the apoptosis induced by caspase-8 is inhibited by the binding of BSP and αvβ3.