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

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 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.

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 study the biodistribution of anti-HER-2/neu monoclonal antibody Herceptin labeled by 131I(131I-Herceptin) in healthy KM mice and nude mice bearing human ovarian cancer xenografts and radioimmunoimaging (RII) of the nude xenografts-bearing mice.Methods 131I-Herceptin was prepared using Iodogen method.The labeling efficiency, radiochemical purity, stability and immunocompetence were measured.The percentage activity of injection dose per gram of tissue (%ID/g) and the radioactivity ratio of tumor to non-tumor tissue (T/NT) were calculated for each time point.The optimal time for imaging was investigated by comparing the 131I-Herceptin SPECT for the nude mouse models bearing ovarian cancer xenografts at different time points.Results The labeling efficiency and radiochemical purity of 131I-Herceptin were 89.8% and 98.4%, respectively.The labeling was stable and had good immunocompetence.131 I-Herceptin was cleared rapidly mainly through liver, spleen and kidneys, consistent with first order two-compartment model.The uptake of 131I-Herceptin in the tumors bearing human SKOV-3 xenografts was much higher than that in nontumor tissue.The% ID/g was 18.08 in the tumor at 24 h post injection.The T/NT ratio increased with time and was 27.27 at 72 h post injection.The tumors in nude mice bearing SKOV-3 xenografts could be visualized on 131I-Herceptin SPECT imaging 2 h post injection; definitely identiffed 48 h post injection and the radioactivity ratio of tumor to contralateral tissue was 11.44 at 120 h post injection.However, the tumor in nude mice bearing HO-8910 xenografts did not show abnormal uptake of 131 I-Herceptin at each time point.Conclusions 131 I-Herceptin is a good radiopharmaceutical targeting SK-OV-3 xeuografts and it may be useful in imaging carcinoma of ovary and target therapy of its metastases with high HER-2/neu expression.

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.

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.

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.