Objective To discuss how to deal with the problems of LEEP electro-scalpel that are caused by the burn of negative electrode loose and the electric shock of leakage current.Methods The system's security was enhanced by insulation,power and software control and it was also evaluated.Results The electric leakage of LEEP electro-scalpel was acceptable and the preventive support of burn was effective.Conclusion It is proved by the synthetical evaluation that the device's secure design is reliable.

LEEP electro-scalpel is a type of cervical disease device. It belongs to the high frequency electro-knife in medical apparatus category. There is no domestic LEEP electro-scalpel at present. It is significant and important for the study group to resolve the safety and EMC problem and develop domestic LEEP electro-scalpel which is safe, stable and competitive in price while has the same surgery effect as the imported one.

Aim To investigate the role of peripheral benzodiazepine receptor in rat cardiac mitochondrial permeability transition.Methods The isolated rat cardiac mitochondria were incubated with different doses(50,100,200 ?mol?L-1) of PBR antagonist 1-(2-chlorophenyl-N-methyl-1-methylpropyl)-3-isoquinolinecarboxamide (PK 11195). In additional group(CsA group), 5 ?mol?L-1 cyclosporine A (CsA), an inhibitor of MPT was added 5 minutes before the addition of 100 ?mol?L-1 PK 11195. Negative control group(Con group) was given none treatment. Positive control group(Ca2+ group) was given 150 ?mol?L-1 CaCl2. The absorbanceat 520 nm(Abs 520 nm) was monitored with a split-beam spectrophotometer at 30℃ for 10 min. The mitochondrial ultrastructure was assessed by transmission electron microscopy. Mitochondrial cytochrome C release was demonstrated by Western Blotting.Results PK11195 triggered large-amplitude mitochondrial swelling in a dose dependent manner(vs Con group,P

Objective To study ultrasonic parameters characteristics of lactation mastitis,breast cancer by using color doppler flow imaging(CDFI)and ultrasonic elastography(UE),and to explore its clinical value.Methods Retrospective analysis was conducted in the clinical data of 85 female patients (a total of 85 breast lesions).All patients were confirmed by pathology,and according to the results of pathology,they were divided into non lactation mastitis group(group NLM,a total of 28 cases),breast cancer group(group Ca,a total of 57 cases).All patients accepted CDFI and UE before treatment,and compared ultrasound parameters between the two groups.Results The proportions of class 0~I(71.43%)and RI<0.7(82.14%)in group NLM were significantly higher than those in group Ca,while the proportions of classⅡ ~Ⅲ(28.57%)and RI≥0.7 (17.86%)were significantly lower than 75.44%,84.21% in group Ca(χ2 =17.185,35.217,all P<0.05).The UE ratings(1.75 ±0.97)and the strain rate ratio (1.64 ±0.83)in NLMgroup were lower than (4.19 ±0.74),(5.03 ±1.08)in group Ca(t=-12.873,-14.623,all P<0.05).The accuracy of CDFI +UE(89.41%)was higher than the accuracy of UE(76.47%)or the accuracy of CDFI(67.06%)(χ2 =12.337,P<0.05).Conclusion CDFI,UE have a certain diagnosis ability for non lactation mastitis and breast cancer,combining both can obtain better diagnostic value,which is worthy of clinical application.

Aim To establish a method to quantitatively determine cardiolipin content and investigate effects of age and ischemia on cardiolipin in isolated rat hearts.Methods Male SD rats, 4 months old,12 months old and 24 months old were used throughout the experiment. Each had 24 ones,which were randomly distributed to control,ischemia 30 min and ischemia 40 min groups(n=8). Control group hearts were perfused for a total of 70 min and ischemia groups hearts were perfused for a 20 min equilibration period,and then underwent 30 or 40 min of no-flow ischemia followed by reperfusion for a 20 min period, respectively.Cardiolipin was quantitatively determined by high-performance liquid chromatography following extraction of lipids from subsarcolemmal mitochondria(SSM)and interfibrillar mitochondria(IFM).Results Under the condition of HPLC,the retention time of cardiolipin was 13.092 min. The standard curve was Y^ (?V?s) =20 877 455X(mmol?L-1)-352 028(r=0.9993) with a validated quantitation range of 0.2～3.2 mmol?L-1,and the lowest concentration of detection was 0.005 mmol?L-1 (S/N=3). At the concentration of 0.2,0.8,3.2 mmol?L-1,absolute recovery rates ranged from 0.8505 to 0.9519 and relative recovery rates ranged from 0.9898 to 1.0429. The RSD of the precision within-day and between-day was less than 0.11. Cardiolipin content in SSM of nonischemic group of 24 months old rats was lower than that of both 4 months old and 12 months old, but no differences among them were observed in IFM.In all age rats, compared with control group,cardiolipin contents of SSM from ischemia 30 min and ischemia 40 min groups were lower, and effects of ischemia 40 min are not pronounced; there were no differences in CL contents of IFM among three groups.Conclusions The assay method was shown to be sensitive, suitable and accurate for determination of cardiolipin located in mitochondria from perfused hearts of rats. Myocardial ischemia and older age decreased cardiolipin content in SSM, and had no remarkable effects on cardiolipin in IFM.

Objective To investigate the impact of reduced glutathione(GSH) on the prolifera- tion,oxidative stress and transforming growth factor?1(TGF-?1) expression of human hepatocytes and hepatic stellate cells(HSCs)(LX-2 cell line).Methods Human hepatocytes and HSCs were incubated with various concentrations of GSH(0.5—50 mmol/L or 0.5—10 mmol/L).The effects of GSH on the proliferation of hepatocytes and HSCs were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphennyhera- zolium bromide colorimetric assay.Human hepatocytes and HSCs were co-cultured with GSH and ferric nitrilotriacetic acid,superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were detected.HSCs were incubated with high(5.0 mmol/L),media(2.5 mmol/L) and low (0.5 mmol/L) concentrations of GSH,the expressions of TGF-?1 mRNA and protein were detected by ELISA and real- time PCR.Results In concentration ranged from 2.5 to 10 mmol/L,the GSH could promote the pro- liferation of hepatocytes but no HSCs,significantly increased the activity of SOD and decrease the con- tents of MDA in hepatocytes and HSCs,and inhibited the expression of TGF-?1 in HSCs.Conclusions GSH can not only promote the proliferation of hepatocytes,but also protect hepatocytes and HSCs from oxidative stress,and inhibit the secretion of TGF-?1 in HSCs.GSH may play a role in hepatocellular protection,antioxidation and anti-fibrosis.

Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.

AIM:To study the telomere maintenance mechanism in mesenchymal stem calls(MSCs).METHODS:MSCs were isolated from healthy human bone marrow by their adherence to plastic and then were checked with CD14-FITC,CD45-FITC,CD44-FITC,HLA-DR-FITC,CD34-PE,CD29-PE and CD166-PE.Telomere length and ECTR DNA in MSCs were detected by Southern blotting.The localization of TRF1 and promyelocytic leukemia(PML)in MSCs were detected with immunofluorescence staining.TRAP protocol was performed to detect the telomerase activity in MSCs and MSCs-derived adipocytes.Western blotting and TRAP protocol were applied to measure telomerase activity of MSCs,which were synchronized by serum starvation and aphidicolin treatment.RESULTS:The telomere in length seemed shorter and relatively more homogeneous in MSCs and HeLa cells than that in WI-38-2RA cells.TRF1 did not concide with PML nuclear body in MSCs and HeLa cells while it exclusively did in WI-38-2RA cells.ECTR DNA was negative in MSCs and HeLa cells but positive in WI-38-2RA cells.Telomerase was negative in MSCs but it was positive in MSCs-derived adipocytes detected by TRAP.Moreover,a cell cycle-dependent expression profile of telomerase was found in MSCs when they were synchronized by serum starvation and aphidicolin treatment.Untreated MSCs expressed very low level of telomerase probed by Western blotting with 2C4 mAb,but the telomerase level had significantly increased when these cells were trapped in S phase.CONCLUSION:The telomere of MSCs is maintained by telomerase pathway instead of alternative lengthing of telomere(ALT)and the level of telomerase expression is associated with cell cycle stage.[

Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis, pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using immunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.

Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis, pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using immunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected.