Objective To investigate the effect of sufentanil on anoxia/rexogenation (A/R)-induced injury to H9c2 cells incubated in high-glucose culture medium. Methods The H9c2 cells were cultured in low-glucose DMEM/F12 culture medium supplemented with 10% fetal bovine serum. The cells were seeded in 96-well or 6-well plates and randomly divided into 5 groups ( n = 12 wells each): normal glucose control group (group NC),high-glucose control group (group HC), high-glucose + A/R group (group HA/R), high-glucose + sufentanil +A/R group (group HSA/R), high glucose + naloxone + A/R group (group HNA/R). The cells were exposed to 95 % N2-5 % CO2 in an incubator at 37 ℃ for 3 h followed by 3 h reoxygenation. In group NC, the cells were incubated in low-glucose culture medium for 48 h. In group HC, the cells were incubated in high-glucose culture medium for 48 h. In group HA/R, the cells were incubated in high-glucose culture medium for 48 h before anoxia.In group HSA/R, after the cells were incubated in high-glucose culture medium for48 h, sufentanil was added to the culture medium with the final concentration of 10-9 mol/L at 15 min before anoxia. In group HNA/R, after the cells were incubated inhigh-glucose culture medium for 48 h, naloxone was added to the culture medium with the final concentration of 10-6 mol/L, 10 min later sufentanil was added with the final concentration of 10-9 mol/L at 15 min before anoxia. The cell viability was measured by MTT assay. The amount of lactic dehydrogenase (LDH) released in the supernatant and superoxide dismutase (SOD) activity were measured. Results The cell viability and SOD activity were significantly lower, while the amount of LDH released was significantly higher in the other groups than in group NC, in groups HA/R and HNA/R than in group HC, and in group HNA/R than in group HSA/R (P ＜ 0.01 ). The cell viability and SOD activity were significantly higher, while the amount of LDH released was significantly lower in group HSA/R than in group HA/R ( P ＜ 0.01 ). There was no significant difference in the parameters mentioned above between group HNA/R and group HA/R ( P ＞ 0.05). Conclusion Sufentanil can attenuate A/R-induced injury to H9c2 cells with high-glucose incubation, and the mechanism may be related to the activation of opioid receptors.

Objective To investigate the effects of Triptolide on apoptosis of cultured rat mesangial cells treated by TGF-β1 and the role of Smac in this process. Methods The mesangial cells were pre-treated with different concentrations of Triptolide for 24 hours, then stimulated with TGF-β1 for 24 hours. Apoptotic cells were detected by TUNEL assay. Smac transcription level was determined by Real time-PCR analyses. Smac expression level was assessed using Western blot anal-yses. Localization of Smac was shown by confocal fluorescence microscopy. Results Compared with control group, TGF-β1 inhibited apoptosis and Smac transcription and expression in rat mesangial cells. By contrast, Triptolide promoted mesangial cells apoptosis. In Triptolide groups, Smac mRNA and protein levels were up-regulated. Additionally, in normal and TGF-β1 groups Smac protein was mainly localized in mitochondriawhile in Triptolide groupit was mainly localized in cytoplasm and nucleus with increased fluorescence intensity. Conclusion Triptolide could promote the effect that TGF-β1 inhibited apop-tosis of mesangial cells, through both up-regulation the expression of Smac and stimulating it translocation from mitochon-dria to cytoplasm and nucleus.

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Objective To analyze the voluntary blood donors of Shaoguan city in recent 10 years HIV infection status and distri‐bution characteristics ,and to take effective prevention and control measures ,in order to reduce the risk of HIV infection through blood transfusion .Methods HIV screening test in the early by the reagent of two different manufacturers ,from 2005 to 2014 vol‐untary blood donors specimens of 265 771 doses of blood donation in Shaoguan ,and early screening positive specimens confirmed by Center for Disease Control of Shaoguan .Results The voluntary blood donors of Shaoguan in the 10 years that initial screening test anti‐HIV positive were 310 cases ,and 35 cases confirmed positive;The confirmed positive rate was 1 .317/10 000;The HIV infec‐tion rate in 2012 was highest (2 .867/10 000) and there were 9 cases ,which was significant difference (P<0 .05) .The district of the maximum rate was New Fengxian ,which was 5 .369/10 000 ,and different districts had significant difference (P<0 .05) .The epidemiology survey about HIV infections found that male were 30 cases(85 .71% ) ,which was significantly more than women;By age distribution ,most HIV infections were 18 to 35 years old young adults ,which accounted 26 cases(74 .29% ) .Married and un‐married people infected respectively accounted 28 .57% and 71 .43% .The occupations of HIV infections were scattered with farm‐ers ,staffers ,the service industry ,cadres in the majority .The HIV infection rate with low education was relative higher ,which had some relationship to educational level .The residents and the non‐residents infected respectively accouted 71 .43% and 28 .57% ;Blood donors for the first time of HIV infections were 24 cases (68 .57% ) .Blood donors for the second time of HIV infections were 11 cases (31 .43% );Other virus co‐infection respectively were 2 HBV infections ,1 HCV infection and 3 syphilis infections .The route of transmission was mainly through sexual transmission that was mainly heterosexual transmission ,which was 33 cases (94 .58% ) .Conclusion The voluntary blood donors of Shaoguan with HIV is on the rise ,and give priority to with male youths spread through heterosexual sex .It should be pay high attention to by the government ,society and blood stations ,and strengthen the propaganda work of AIDS prevention and control checks ,and minimize the risk of HIV transmission by blood transfusion in or‐der to guarantee the safety of clinical blood transfusion .

BACKGROUND:A group of nuclease-like proteins were previously purified from Eisenia foetida tissues, exploring primary structures of these proteins wil help to uncover basic structure characteristics of them and provide foundations for the study addressing the relationship of their structures and functions.
OBJECTIVE:To explore primary structures of nuclease-like proteins EWD1 and EWD2.
METHODS:Edman degradation method was used to sequence the N-terminal amino acids of EWD1 and EWD2, acid hydrolisis method was used to analyze amino acid compositions of EWD1 and EWD2, LC-MS/MS was used to analyze some peptide sequences within the proteins, and MALDI-TOF-MS was used to calculate the number of the disulfide bonds and the contents of polysaccharides.
RESULTS AND CONCLUSION:Among the amino acid compositions in EWD1 and EWD2, the sum contents of aspartate and asparagines were the highest (al nearly 10%), the contents of hydrophobic amino acids were also high, and the contents of cysteine was low. The EWD1 and EWD2 had similar amino acid compositions with other nucleases. Edman degradation results showed that, the N-terminal sequences of the large subunit of EWD1 were in turn as fol ows:D, E, W, V, Y, P;the N-terminal sequences of EWD2 were as fol ows:L, L, G, P, Y, K, P, K, C. The results of LC-MS/MS indicated the two proteins were novel proteins;MALDI-TOF-MS results showed that 8 cysteine residues formed 4 disulfide bonds in EWD1, 6 cysteine residues formed 3 disulfide bonds in EWD2. EWD1 and EWD2 were al glycoproteins, the content of polysaccharides was 17.3%in EWD1 and 15.6%in EWD2.

Aim To observe the protective effect of quercetin on rat cardiomyocyte apoptosis induced by adriamycin and explore its possible mechanism.Methods Cultured neonatal rat cardiomyocytes were randomly divided into six groups:normal control group, adriamycin group,quercetin control group, adriamycin+quercetin(25,50,100 ?mol?L-1)groups. The activity of LDH was detected by chromatometry, the cardiomyocyte viability was measured by MTT, the ultrastructure of cardiomyocyte was observed by electron microscope, the expression of protein Bcl-2 and Bax was analyzed by immunocytochemical, and the mRNA and protein of caspase-3 were detected by RT-PCR and Western blot respectively.Results Compared with the control group, the activity of LDH was increased but the viability of cardiomyocyte was decreased; the expression of Bax and caspase-3 was up-regulated while Bcl-2 was down-regulated in ADR group.Compared with ADR group, the above changes were lightened in adriamycin+quercetin groups. But the quercetin control group, in which cultured myocardial cells only exposed to quercetin without ADR, had no obvious changes.Conclusions Quercetin significantly inhibits the apoptosis induced by ADR in the cultured myocardial cells. Its mechanism is involved in the apoptosis-related pathways, including caspase-3, Bax and Bcl-2.

Objective To investigate the risk factors related to failure of proximal femoral nail anti-rotation(PFNA)internal fixation for intertrochanteric fracture of femur.Methods A retrospective research was done on 249 cases with intertrochanteric fracture of femur after PFNA internal fixation.The failure ratios related to sex,fat,fracture pattern,osteoporosis,replace condition in operation and tip-apex distance(TAD)were calculated.And the risk factors related to failure of PFNA were statistically analysed.Results Two hundred and forty-three cases were followed up ranged from 10 months to 3 years after operation.Plain AP and lateral radiographs were obtained at each follow-up patient,13 of 243 cases resulted in failing by PFNA internal fixation.The failure ratios in PFNA internal fixation with fat,unstable fracture,TAD≥25 mm and bad replace condition in operation were 8.5％(12/141),7.8％(12/153),18.5 ％(5/27)and 26.1％(6/23),respectively,which were higher than those in PFNA internal fixation with nonobese,stable fracture,TAD ＜ 25 mm and good replace condition in operation[1.0％(1/102),1.1％(1/90),3.7％(8/216)and 3.2％(7/220),respectively](P＜0.05 or ＜0.01).Conclusions Fat,unstable fracture,TAD≥25 mm and bad replace condition in operation are the relatively independent risk factors related to failure of PFNA internal fixation for intertrochanteric fracture of femur.Furthermore,it will prevent and reduce the ratio of complications to avoid these risk factors when PFNA internal fixation is selected for treatment intertrochanteric fracture of femur.

Objective To explore the effect of the proximal femoral nail antirotation(PFNa) for the treatment of femoral intertrochanteric fracture.Methods 272 consecutive patients with femoral intertrochanteric fracture were treated with PFNa.Results The mean operative time was 55min(range 35 ～95)min.The mean blood loss was 85ml (range:20 ～200)ml.The mean follow-up was 15.6months( range:6 ～36)monthsand achieved bone healing with an average time of 14.5weeks.According to the Harris criterion for evaluation.Conclusion PFNa was more comfortable for human's biomechanism and more favorable for the healing of the fracture as well as the recovery of the hip function.It had the advantage of simple manipulation,fewer trauma,strong fixation,anti-rotation,less osclerotin.PFNa was an ideal treatment for intertrochanteric fracture,specially the unstable intertrochanteric fracture and senile osteoporosis.

Objective To investigate the incidence of diseases in member of rescue teams in the Wenchuan earthquake in Sichuan Province.Method Twenty days afer the Wenchuan earthquake in 2008,5 068 resucing staff of rescue learns as random and cluster sample were investigated and analyzed to find the factors impacting their fitness associated with their characters,diseases,work,and encampment styles.Results Members of rescue teams suffered from the following diseases:insect dermatitis (18.37%),tinea pedis (10.83%),acute upper respiratory infection (10.56%)and solar dermatitis (6.20%).Contrasted by work tasks,staff resucing on the front line had the highest incidence of acute upper-respiratory infection(11.04%) ,logistics workers had the highest incidence of tinea pedis(15.21% ),and more patients of acute gastroenteritis,insect dermatitis,gingivitis emerged from medical personnel group(P<0.05).The encampment sites were devided into three sorts:plain land,hillside and valley.The staff stayed at valley had higher incidence of acute upper respiratory infection (24.90%),insect dermatitis (36.50%) and tinca pedis (20.02%)than those worked at plain land and hillside (P<0.01).The incidence of acute gastroerrteritis(1.26%) ha staff resides hillside is lower than that in valley or plain land (P<0.05).Conclusions In the period of rescue actions in the guake-hit region,the members of rescue teams suffered from famihar diseases such as dermatitis and acute upper-respiratory infection,etc.The incidence of diseases is associated with their work tasks and the topography of cantonment.

Objective To study the effects and mechanisms of molecular targeted drug combination on multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells. Methods Four molecular targeted drugs (HG6-64-1, Dasatinib, Crizotinib, and Sunitinib) were used to treat SK-Hep-1 cells, and the monophasic kinetic analysis curve and two-phase analysis curve were drawn. Western blot analysis was used to detect the effects of the above drugs on key signaling pathways in SK-Hep-1 cells. MTT assay was used to detect the effects of the above drugs and their combination on the proliferation of SK-Hep-1 cells. Results Compared with the monophasic kinetic analysis curve, the biphase analysis curve could better fit the effects of molecular targeted drugs on SK-Hep-1 cells, which predicted that the combination of HG6-64-1, Dasatinib, and MK-2206 could effectively inhibit the proliferation of SK-Hep-1 cells. Conclusion Two-phase kinetic analysis can quantitatively describe the response of multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells to molecular targeted therapy. The combination of HG6-64-1, Dasatinib, and MK-2206 is a potential drug combination for the treatment of hepatocellular carcinoma.