Objective To establish a real-time quantitative reverse-transcription PCR ( qRT-PCR) method for detection of hepatitis E virus ( HEV) of different genotypes based on standard HEV DNA plasmid in order to promote its application in clinical laboratory. Methods Specific primers and probe of HEV were designed based on the conserved open reading frame 3 (ORF3) regions. HEV DNA plasmids were construc-ted and 10-fold serial dilutions of the plasmids were prepared and used as standards to establish one-step qRT-PCR. The established method was compared with HEV antigen, antibody and RT-nPCR assays. Some positive samples were sequenced and analyzed by evolutionary tree. Results The one-step qRT-PCR meth-od for HEV detection in serum or feces samples was successfully establish. It could reach a sensitivity of 25 copies/test and 77. 8% of its results were consistent with those by HEV antigen assay. Nine patients were infected with HEV of genotypes 4a, 4d or 4n as indicated by evolutionary tree. Conclusion The HEV qRT-PCR method based on its standard plasmid is successfully established, which paves the way for commercial-ization of clinical applications.

Lumbar dysfunction closely relates to the weakness or deficiency of lumbar proprioception, and accurate and comprehensive lumbar proprioception test is the important basis of training program formulation, efficacy assessment and prognostic evaluation. Comprehensive lumbar proprioception test includes position sense test, kinesthesia test and vibration sense test. It has been widely used in clinic at abroad, and is rich in test equipments and methods. Domestic research is still at the preliminary stage in this field, and is lack of accurate, objective and unified test methods. According to an overview of relevant literature, this article discussed the lumbar proprioception test methods, the influential factors and reliability of the test, finally, put forward the prospects about the research direction in this field.

AIM:To study the effects and mechanisms of ethanol on chloride channels in poorly differentiated nasopharyngeal carcinoma CNE-2Z cells.METHODS:The effect of ethanol on the cell growth was analyzed by MTT as-say.The technique of whole-cell patch-clamp was used to detect the chloride current .The characteristics of the chloride current were analyzed by using the chloride channel blockers .The siRNA technique was used to analyze the molecular basis of the ethanol-sensitive chloride channels .RESULTS: Under isotonic conditions , the background current was weak and stable.Ethanol at concentrations of 0.17~170 mmol/L activated a chloride current in a concentration-dependent manner (an inverted U-shape), with a maximum effect at the concentration of 17 mmol/L.The currents showed obviously outward rectification and were susceptible to extracellular hypertonicity and the chloride channel blocker , 5-nitro-2-(3-phenylpropyl-amino) benzoic acid ( NPPB) .ClC-3 siRNA obviously decreased the currents activated by ethanol .CONCLUSION:Ex-tracellular ethanol induces chloride currents through activating the ClC-3 chloride channels .

Objective:To explore the effects of caffeine on the prevention of Alzheimer's disease (AD).Methods:Use Ethanol as a solvent to extract the caffeine in tea and then injecting 5％ D-galactose saline solution 1ml/d/kg to establish aging model mice.Divide mice randomly into experimental group (high-dose/low-dosecaffeine),positive control group,negative control group,and normal con-trol group (NS) and injecting appropriate drugs for consecutive four weeks.Test superoxyde dismutase (SOD) and malondialdehvde (MDA) periodically.Take mice's hippocampus and use Western blotting to detect the expression of brain derived neurotrophic factor (BDNF) and extracellular signal-regulated kinasesl/2 (p-ERK1/2).Results:The expression of BDNF and p-ERK1/2,negative control group is less than low-dose experimental group and positive control group (P＜0.01);The p-ERK1/2 expression of injecting D-galactose mice was significantly lower than normal group,negative control group compared weth the normal group,the differencd was significant (P＜0.05).The level of SOD in model group was significantly lower than that in normal control group,high,low dose caffeine group and positive control group (P＜0.01),but the level of MDA is opposite.Conclusions:Caffeine can delay aging process by increasing the level of SOD in aging mice,and enhancing the expression of BDNF and P-ERK1/2.Caffeine does a lot to prevent AD.

Objective To investigate the role of TLR4/NF-κB signaling pathway under the action of TAK-242 in the cardiomyocyte apoptosis after coronary micro-embolism (CME) in rats.Methods Fortyfive rats were randomized (random number) into three groups:sham operation,CME and CME plus TAK242 groups (n =15 per group).CME was induced by injecting polyethylene microspheres (42 μm) into the left ventricle except the sham group.CME plus TAK-242 group was treated with TAK-242 (2 mg/kg) via the tail vein of mice 30 min before CME modeling.Cardiac function was evaluated 6 h after operation.Tissue biopsy was stained with HBFP to measure the size of infarction area.TUNEL assay was used to detect cardiomyocyte apoptosis.Western blot and qPCR were used to evaluate the protein levels and mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3,respectively.Statistical analysis was performed using one-way analysis of variance followed by LSD-t test.Results Compared with the sham group,left ventricular ejection fraction (LVEF) in the CME group was significantly decreased [(68.91 ± 4.12) ％ vs.(84.80 ± 2.51) ％,P ＜ 0.05],and the infarction area (P ＜ 0.05),the apoptosis index [(3.36 ± 0.63) ％ vs.(0.19 ± 0.08) ％,P ＜0.05],the mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3 in CME group were increased significantly (all P ＜ 0.05).Compared with CME group,LVEF in the CME plus TAK-242 group was significantly improved [(75.58 ± 5.01) ％ vs.(68.91 ± 4.12) ％,P＜0.05],and the infarction area [(8.58 ± 2.12) ％ vs.(14.65 ± 4.23) ％,P＜0.05],the apoptosis index [(1.43 ± 0.51) ％ vs.(3.36 ± 0.63) ％,P ＜ 0.05],the mRNA expressions of TLR4,NF-κB p65 and cleaved caspase-3 in CME + TAK-242 group were decreased significantly (all P ＜ 0.05).Conclusions TAK-242 effectively improved CME-induced cardiac dysfunction by regulating TLR4/NF-κB signaling pathway and then reducing the cardiomyocyte apoptosis.

Objective To compare the efficacy of the reforming endoscopic minimally invasive releasing(REMIR) with open procedure for treatment of carpal tunnel syndrome.Methods Senventy patients with unilateral carpal tunnel syndrome were randomly divided into REMIR group and open procedure group,with 35 cases in each group.Kelly's standard,two-point discrimination,operation time and complication occurrence were compared between the two groups.Results All patients were followed-up for 12 months.There was no significant difference in the therapeutic results and in the improvement of two-point discrimination between the two groups (P ＞ 0.05 ).The operation time of REMIR group was significantly shorter than the open procedure group ([ 10.03 ± 1.84] min vs [37.63 ±7.18]min,t =22.210,P ＜0.001 ).And there was no cases with scar tenderness in REMIR group while there was 7 cases in open procedure group.Conclusion Compared with the open procedure,the REMIR method has the same clinical efficacy while with the advantages of causing smaller skin scar and being less time consuming.

Objective To compare the clinical efficacies of Qin’s scalp eight-needle acupuncture versus conventional scalp acupuncture in treating cancer pain. Method Sixty cancer pain patients with clinically or pathologically diagnosed malignant tumors were allocated, using a random number table, to a treatment (Qin’s scalp eight-needle acupuncture) group of 30 cases and a control (conventional scalp acupuncture) group of 30 cases. The treatment group received Qin’s scalp eight-needle acupuncture at selected points every other day and the control group, conventional scalp acupuncture every other day. One course of treatment consisted of 10 days. Result Cancer pain was relieved, analgesic dosage decreased, adverse reactions to analgesics reduced and quality of life raised significantly in the treatment group compared with the control group. Conclusion Qin’s scalp eight-needle acupuncture is effective in treating cancer pain.

AIM:To investigate the type of chloride channel activated by cisplatin in poorly differentiated na -sopharyngeal carcinoma cells (CNE-2Z cells).METHODS:The technique of whole-cell patch-clamp was used to investi-gate the role of Ca 2+in the activation of cisplatin-activated chloride currents and to analyze the effect of hypertonic stress on these currents in CNE-2Z cells.RESULTS:Chloride currents were induced when the cells were exposed to the calcium -free cisplatin solution , showing the similar density to the currents induced by cisplatin with the presence of extracellular cal -cium.However , the latency and the peak time of cisplatin-activated currents in the absence of extracellular calcium were prolonged.The activation of cisplatin-activated chloride currents was insensitive to the depletion of intra-and extracellular calcium.Calcium channel antagonist nifedipine had no effect on the cisplatin -activated chloride currents , while hypertonic solution completely inhibited those currents .CONCLUSION:The cisplatin-activated chloride currents are independent on intra/extracellular calcium .The chloride channels activated by cisplatin are not calcium-activated chloride channels , but are probably volume-sensitive chloride channels .

Objective To investigate the relationship between acute radiation proctitis and radiation dose,volume as well as radiation time,in the process of intensity-modulated radiation therapy (IMRT) for the cervical cancer patients.Methods A total of 51 patients with locally advanced cervical cancer were enrolled from January 2011 to December 2013.Those patients were then classified into grade 1 to 4 groups,according to the RTOG/EORTCtoxicity grading standard.The exposure dose volume and the average dose of rectum under the standard plan were evaluated with dose-volume histogram (DVH).The ANOVA test was used for analyzing Dmax,D mean,D1 cm3,D2cm3,D40 and V40 values of rectum and the average exposure dose of rectum.Results The average time of acute radiation proctitis with clinical symptoms was (23.06 ± 12.01) d after radiotherapy.Dmaxvalues of rectum in grade 2 group was lower than those in grade 3 and 4 groups (F =5.268,P ＜ 0.05).Moreover,D1 cm3 and D2 cm3 values of rectum in grade 1 and 2 groups were also lower than those in grade 3 and 4 groups (F =4.893,4.406,P ＜ 0.05).There was no statistically significant difference between D40 and V40 values.Conclusions The acute radiation proctitis could be frequently found around 20 days during the IMRT for cervical cancer patients.Mild and moderate acute radiation proctitis are more common,while severe acute radiation proctitis is rare.Minimizing Dmax,D1 cm3 and D2 cm3 values of rectum might reduce the incidence of severe acute radiation proctitis in cervical cancer patients receiving IMRT.

BACKGROUND:Bone marrow mesenchymal stem cel s transplantation can inhibit experimental emphysema inflammatory reaction and apoptosis, and has been experimental y confirmed to treat severe lung function impairment. OBJECTIVE:To explore the inhibitory effects of bone marrow mesenchymal stem cel s transplantation via different ways on inflammatory reaction and apoptosis due to experimental emphysema. METHODS:Female Wistar rats were randomly divided into control group, intravenous group and endotracheal group fol owing model establishment using fumigation plus intratracheal instil ation of porcine pancreatic elastase. In the latter two groups, bone marrow mesenchymal stem cel s from male rats were injected via the tail vein and the trachea, respectively. In the control group, rats were given PBS via he tail vein and trachea. At 14 days after transplantation, pathological changes of rat lung tissues were observed, cel apoptotic index in alveolar wal cel s and tumor necrosis factorαlevel in the bronchoalveolar lavage fluid were detected. RESULTS AND CONCLUSION:Compared with the control group, in the intravenous and endotracheal groups,the pathological changes of lung tissues were relieved, tumor necrosis factorαlevel and apoptosis index were reduced significantly (P<0.01);but there were no differences between the intravenous and endotracheal groups (P>0.05). These findings indicate that bone marrow mesenchymal stem cel s transplantation via the tail vein and trachea both can exert obvious therapeutic effects on emphysema. Moreover, cel transplantation via the tail vein is more convenient and easier than that via the trachea in the treatment of emphysema.