BACKGROUND: Gene therapy is a popular topic in domestic and overseas studies on biological therapy for brain tumor.OBJECTIVE: By using a newly constructed eukaryotic expression vector of pCR3-TK, the effect of the HSV-TK/ACV system on the proliferative activity of human glioma cells was investigated.DESIGN: Experimental study based on cells.SETTING: Department of neurosurgery and department of oncology in a university hospital.MATERIALS: The study was conducted at the National Key Laboratory of Veterinary Biotechnology of Harbin Veterinary Research Institute from January to April in 2004. The eukaryotic expression vector of pCR3-TK was constructed by the author. The TJ905 strain was a gift from professor Pu Pei-yu, who worked in the Neurology Institute of Tianjin city. The nontransfected cells and the cells transfected with pCR3-Uni vector were set as controls.METHODS: By using Lipofectamine(a cationic liposome), the pCR3-Uni vector and the recombinant pCR3-TK plasmid(inserted with HSV-TK gene)were transfected into the human glioma cell strain-TJ905. Then the positive clones were picked out and were given ACV(50 mg/L) . Totally 72 hours later, the cover slips were collected and silver staining for nucleolus organizer regions(AgNORs) was performed.MAIN OUTCOME MEASURES: After the ACV treatment and AgNORs staining, the numbers of silver-stained granules in TJ905 cells with or without transfections were counted respectively.RESULTS: In those cells transfected with HSV-TK gene, after ACV treatment, a significant decreasing in proliferative activity could be observed, and the average numbers of the silver-stained granules in cells transfected with pCR3-Uni or pCR3-TK were 14.33 and 6.67 respectively( P ＜ 0.01).CONCLUSION: As an easy-to-operate method, AgNOR counting is helpful for the studies on the proliferative activity of cells and the investigations into the potential anti-tumor mechanism of the HSV-TK/ACV system.

Objective To quantify the concentration of peripheral blood plasma neutrophil gelatinase-associated lipocalin (NGAL) in patients with acute pancreatitis (AP) and to explore its value in assessment of the severity of AP.Methods From June 2011 to March 2012,83 patients with AP were selected,among those 43 cases were mild acute pancreatitis (MAP) and 40 were severe acute pancreatitis (SAP).The control group included 30 healthy individuals.The peripheral blood of patients with AP and healthy controls was collected,and plasma was isolated after centrifuged.The concentration of NGAL in plasma was detected by enzyme-linked immunosorbent assay (ELISA).The t-test was performed for comparison between groups.The correlation between the concentration of NGAL in plasma and clinical parameters of AP was analyzed by Spearman rank order correlation analysis.The diagnosis value of the concentration of NGAL in SAP was evaluated by receiver operating characteristic (ROC) curve and area under the curve (AUC).Results The concentration of plasma NGAL in AP group ((10.30± 5.97)nmol/L) was higher than that in healthy control group ((1.94±1.35) nmol/L) and the difference was statistically significant (t=11.924,P＜0.01).The concentration of plasma NGAL in SAP group ((14.61 ±5.28) nmol/L) was higher than that in MAP group ((6.27±-3.09) nmol/L) and healthy control group,the differences was statistically significant (t=8.677 and 14.539,both P＜0.01).The concentration of plasma NGAL of AP patients was positively correlated with acute physiology and chronic health evaluation-Ⅱ score,Ranson score,bedside index for severity in acute pancreatitis score,computed tomography (CT) severity index,C-reactive protein,white blood cells and the days of hospitalization (r=0.651,0.556,0.514,0.620,0.320,0.458 and 0.346,all P＜0.05).The area under the ROC curve of plasma NGAL concentration in diagnosis of SAP was 0.926 (95％CI:0,870-0.983).The cutoff value of plasma NGAL level in diagnosis of SAP was 8.44 nmol/L.The sensitivity and specificity was 87.5 ％ and 88.9％,respectively.Conclusions Plasma NGAL level is correlated with the severity of patients with AP.NGAL may be one of the markers for the early diagnosis of SAP.

Acupuncture Therapy ; Animals ; Female ; Humans ; Polycystic Ovary Syndrome ; therapy

Objective:To evaluate the expression of apoptosis related gene Fas Ligand(FasL) in human hepatocellular carcinoma(HCC) cells HepG2 and its significance in apoptosis.Methods:The recombinant eukaryotic expression plasmid pcDNA3.1hisB- FasL was transfected into HCC cells HepG2 by lipofection, and then soluble FasL was examined in the supernatant of culture cells by EIA, FasL expression in HepG2 cells were detected by immune histochemistry. After stained by annexin V and propidium iodine, cells were passed throw flow cytometer and examined by fluorescence microscope and sym-focus laser scaning microscope.Results:In comparison with untransfected cells,the soluble FasL could be detected in the supernatant of transfected cells, Fas L can be expressed in the membrane and cytoplasm of transfected cells. The apoptotic cell rate in transfected cells was 36.30%, as the control, untransfected cells was 11.53%.Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine stain.Conclusion:This supports a novel pathway of HCC cells were apoptotic itself via the CD95-CD95 ligand system without involvement of immune cells.

Objective:To study the effect of intraperitoneal hyperthermic perfusion chemotherapy combined with venoclysis on postoperative gastric cancer.Methods:140 postoperative patients with gastric cancer were divided into IPHPC group(n=72) and chemotherapy of venoclysis group(n=68).The intraperitoneal hyperthermic perfusion chemotherapy and chemotherapy of venoclysis were applied for the patients in IPHPC group at one time.Postoperative complications,toxicity,survival rate and peritoneal recurrence rate in two groups were observed.Results:There was no sharp difference in the postoperative complications and toxicity between the two groups.The 3,5 year-survival rate were 86.1%,60.2% and 58.3%,29.4% in IPHPC group and CV group.The 3,5 year-peritoneal cavity recurrence rate were 5.6%,20.6%and 27.8%,53% in IPHPC group and CV group.There were significant differences in survival rate and peritoneal recurrence rate between the two groups.Conclusion:Intraperitoneal hyperthermic perfusion chemotherapy combined with venoclysis can prevent recurrence and metastasis effectively,and improve the survival rate and quality of life of postoperative gastric cancer patients.

Objective To investigate the effect of pelvic floor muscle exercise on perimenopausal female sexual dysfunction. Methods Forty six patients with perimenopausal female sexual dysfunction received pelvic floor muscle exercise. Sexual function and pelvic floor muscle strength were compared between pre-and post-exercises. Result The patients′pelvic floor muscle strength and sexual function after exercises were significantly better than those before the exercises (P<0.05). Conclusion Implementation of the pelvic floor muscles exercise in the patients with perimenopausal sexual dysfunction can strengthen pelvic floor muscle and thus alleviate their sexual dysfunction.

Objective:To explore the effects of LYN on the biological behaviors of gastric cancer cells, such as migration and invasion, and analyze the regulatory mechanism of Wnt/ β-catenin signaling. Methods:Gastric and paracancerous tissue samples from gastric cancer patients admitted to the Affiliated Hospital of Chengde Medical University from Oct. 2017 to May 2018 were selected to detect the expression level of LYN in clinically collected gastric cancer and paracancerous tissue samples by qRT-PCR.Gastric cancer AGS cells were used as experimental subjects, and shRNA-LYN was transfected into gastric cancer AGS cells using Lipofectamine 2000 as experimental group (Sh-LYN group), and untransfected cells as control group (NC group). LYN mRNA expression in AGS cells before and after transfection was detected by qRT-PCR.Transwell detected cell migration and invasion. Cell apoptosis was detected by TUNEL.Western Blot analysis was performed to detect proteins related to the Wnt/β-catenin signaling pathway and to study its regulatory mechanism.Measurement data were expressed as ( Mean± SD), and independent sample t test was used for comparison between the two groups. P<0.05 was considered statistically significant. Results:LYN mRNA expression was significantly up-regulated in gastric cancer tissues ( t=11.04, P<0.001).Sh-LYN transfected gastric cancer cells could make LYN protein expression lower than that of the normal control group( F=961.26, P=0.016).After transfection, migration and invasion of cells in the sh-LYN group were inhibited, and the apoptosis rate of cells in the sh-LYN group was significantly higher than that in the normal control group ( t=3.437, 4.450, P=0.011, 0.026) .Western Blot results: sh-LYN reduced the expression of Wnt3 and β-catenin significantly when compared with the normal control group.The detection of downstream proteins showed that after LYN knockdown, the expression of EMT epithelial labeled E-cadherin was increased, and EMT mesenchymal labeled N-cardherin and vimentin were down-regulated. The other related proteins snail 1 and snail 2 were also down-regulated. Conclusion:LYN knockdown inhibits migration and invasion of gastric cancer cells and induces apoptosis in gastric cancer cells, a process associated with the Wnt/ β-catenin signaling pathway.

Objective To investigate the common sites and risk factom of reoperation for bleeding after coronary artery bypass grafts(CABG).Methods During Jan 2000 to July 2011,2765 CABG procedures hed been done in our hospital,including 874 cases with CABG by cardiopulmonary bypass,1891 cases with off-pump coronary artery bypass grafts (OPCAB),105 cases combined with other procedures (cardiac valve operation,resection of ventricular aneurysm,et al),216 cases applied intra-aortic balloon pump (LABP)).67 patients needed reoperation for bleeding,the morbidity is 2.4％.Factors that might affect the result of reoperation were analyzed by single-factor analysis.Results The following factors were related to high rates of reoperation (P ＜ 0.05 ):the internal mammary artery (IMA) used,anfithrombotic medication discontinued less 5 days before elective surgery,concurrent hypertension,combined with other surgery procedures and ages ＞70 years.It was not signifieantly related to CABG or OPCAB,ff use of IABP,ff concurrent diabetes,number of grafts inserted.Major sources of bleeding found in 48 patients were the IMA and its vessels bed,vein grafts,sternum,aortic faucet,midriff,thymus glrard bed vessels.Conclusion Prepared satisfactorily before operation,avoid risk factors can reduce reoperation for bleeding,but the most important is the operation carefully during operation of hemostasis.

Objective To evaluate the performance of the Sino System for Coronary Operative Risk Evaluation ( SinoSCORE) on in-hospital mortality and postoperative complications in patients undergoing coronary artery bypass grafting (CABG) in a single heart center. Methods From January 2007 to December 2008, clinical information of 201 consecutive patients undergoing isolated CABG in our hospital was collected. The SinoSCORE was used to predict hospital mortality and major complications[sternal wound infection, postoperative renal failure, multiple organ dysfunction syndrome, perioperative intra-aortic balloon pumps ( IABP), etc.]after CABG among our study participants, which was initially designed as CBAG operative risk scoring system and included 11 risk factors. We estimated the predictable capability of SinoORE model by the means of analysing the calibration and discrimination characters of this risk scoring system. Calibration was evaluated with the method of Hosmer-Lemeshow goodness-of-fit test. Discrimination was tested by determining the area under the receiver operating characteristic (ROC) curve. The optimal cut-off point for SinoSCORE predicting major complications was obtained by the Youden index. Results Of all our study patients, the observed in-hospital mortality was 1.99% (4/201). The overall mean baseline age was ( 63.3 ± 9.2 ) years and 24.4% ( 49/201 ) were female. The predicted mortality cakulated by the SinoSCORE was 2.88% which was slightly higher than the actual mortality. SinoSCORE model slwed very high discriminatory ability and the good calibration power in predicting in-hospital mortality: Hosmer-Lemeshow calibration test:x2 =4. 304, P =0.744 and area under ROC was 0. 81 (95% CI: 0.687 -0.932). As for the major postoperative complications after CABG,SinoSCORE model still achieved a satisfactory performance with the good predictive value for the main complications risk evaluation such as postoperative renal failure, multiple organ dysfunction syndrome and IABP. Hosmer-Lemeshow: P =0.75, P =0. 75, P = 1.00; Areas under ROC respectively at was 0. 768 ( 95 % CI: 0.613 - 0. 924 ). 0. 832 ( 95 % CI: 0. 732 - 0. 932 )and 0. 737 (95% CI: 0. 607 -0. 867 ). The optimal cut-off points for SinoSCORE model predicting each of the above three major postoperative complications was ultimately determined to be 4.5. Conclusion In our patient database, the SinoSCORE model proved a good preoperative risk model in predicting both in-hospital mortality and major complications after CABG, which provides a realistic estimation of hospital death and post-operative complications risk for patients undergoing CABG. Sinoscore model is a suitable operative risk estimation system for Chinese CABG patients.

Objective To retrospectively analysis whether the existence of deep vein thrombosis(DVT) in chronic thromboembolic pulmonary hypertension (CTEPH)) has an influence on the efficacy of pulmonary thromboendarterectomy.Methods The data of 75 patients with CTEPH having undergone PTE in Beijing Chaoyang Hospital from Sep 2004 to Mar 2016 were retrospectively reviewed.We classified them into two different groups which were DVT (+)group (32) and DVT (-) group (43) according to the present of DVT.Clinical data,hemodynamics,blood gas analysis and so on of both groups were compared.Results There were 1 patient died after PEA surgery in DVT(+)group and 7 patients died after PEA surgery in DVT (-) group(3.1％ vs 16.3％,P =0.033).The incidence of severe pulmonary reperfusion injury and the neurological disorders postoperatively in DVT(-) group was significantly higher than that in DVT(+) group (46.5 ％ vs 21.9％,P =0.028;32.6％ vs 12.5％,P =0.044).Compared with DVT(+) group,the content of C-react protein (CRP) was higher in DVT (-) group [(0.9 ± 0.7) mg/dl vs (0.5 ± 0.4) mg/dl,P =0.005].Furthermore,DVT(-) group had a significantly greater proportion of Jamieson Ⅲ or V than DVT(+) group,while the cardiopulnonary bypass time [(289.8 ± 54.3) min vs (259.8 ±45.5) min,P=0.014],aorta cross clamp time[(137.3 ±31.6) min vs(119.5±29.3) min,P=0.015] and deep hypothermia circulatory arrest were significantly longer in DVT(-) group than in DVT(+) group.Postoperative hemodynamics and blood gas index were obvious improved in both groups,but the postoperative pulmonary artery systolic pressure (PAPS) [(59.6 ± 17.9) mmHg vs (48.5 ± 9.7) mmHg,P =0.001],the pulmonary vascular resistance (PVR) [(30.44 ± 22.97) kPa · S · L-1 vs (18.34 ±8.09) kPa · S · L 1,P =0.002] in DVT(-) group were significantly higher than those in DVT(+) group.In addition,the incidence of residual pulmonary hypertension in DVT(-) group was significantly higher than in DVT(+) group.Conclusion Pulmonary thromboendarterectomy for patients with CTEPH results in significant pulmonary hemodynamic improvement with favorable outcomes of heart and lung function.The recovery of the PAPS,PVR in patients with DVT are significantly better than those in patients without DVT,and the incidence of postoperative complication is significantly lower in patients with DVT.