In this study, we use retroviral vectors, LXSN (containing internal promoter) and GCEN(containing internal ribosomal entry site,IRES), which cany ?galactosidare gene (LacZ) and neomycin phosphotransferase gene (Neo_R) as double markers, to establish two cell lines, LG-3T3 and GG-3T3. The sensitivities of the two cell lines for detecting replication-competent retroviruses(RCR) in marker rescue test were compared. Our results indicated that both cell lines can be used in marker rescue test, the sentivity of GG-3T3 was 100 times higher than that of LG-3T3.

Objective To evaluate the effect of isoflurane on the expression of matrix metalloproteinase 2 (MMP-2) in human tongue cancer cell.Methods Tea8113 cells at the logarithmic growth phase were seeded into 96-well plates (200 μl/hole) with the density of 5 × 103/ml,6-well plates (5 ml/hole) with the density of 1 ×105/ml,culture dishes (5 ml/dish) with the density of 5 × 105/ml or Transwell chamber (200 μl cell suspension for upper chamber with a density of 5 × 105/ml,10 ％ blood serum medium 500 μl for lower chamber).The cells wererandomly divided into 3 group (n =30 each):control group (group C),2％ isoflurane 2 h group (group Ⅰ1) and 2％ isoflurane 4 h group (group Ⅰ2).2％ isoflurane was added to the culture medium and the cells were incubated for 2 and 4 h in groups Ⅰ1 and Ⅰ2,respectively.The cell viability was detected by MTT assay.The cell apoptosis was assessed by flow cytometry.The migration and invasion of the cells were measured by cell wound scratch assay and Transwell chamber assay,respectively.The expression of MMP-2 and MMP-2 mRNA was detected by immunocytochemistry and RT-PCR,respectively.Results The cell viability and migration of cells were significantly increased,the apoptotic rate was decreased,and the expression of MMP-2 and MMP-2 mRNA was up-regulated in groups Ⅰ1 and Ⅰ2,and the invasion of the cells were increased in group Ⅰ2 as compared with group C (P ＜ 0.05).Compared with group Ⅰ1,the cell viability and migration and invasion of the cells were significantly increased,the apoptotic rate was decreased,and the expression of MMP-2 and MMP-2 mRNA was up-regulated in group Ⅰ2 (P ＜0.05).Conclusion Isoflurane enhances the migration and invasion of the cancer cells by up-regulating the expression of MMP-2 in human tongue cancer Tea8113 cells.

Objective To investigate the expression of apoptosis-related neuclosome assembly protein-1(NAP-1) gene in related brain region of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP) treated mice and beta-amyloid treated rats. Methods The animal models with central nervous system degeneration were established by consecutive administration of MPTP to C57BL mice and injection of beta-amyloid to brain ventricular of SD rats. The RNA in striatum and substantia nigra of mice and the RNA in cortex and hippocampus of rats were extracted. The levels of NAP-1 mRNA in these samples were estimated by RT-PCR. Results For mice, the expression of NAP-1 were decreased after MPTP treatment in substantia nigra, while its expression showed no significant change in striatum. In contrast, no effect on NAP-1 gene expression by beta-amyloid injection can be detected in cortex and hippocampus of rats. Conclusions The apoptosis of dopaminergic neurons in nigrostriatal pathway after MPTP treatment may be related to the expression of NAP-1 gene. But the mechanism of neurodegeneration after beta-amyloid injection in rats may be different.

0.05],while the RDW in CHD patients was significantly increased[(13.2?0.8)%,both P

This study investigated whether the curative effect of short-pulse gastric electrical stimulation (GES) on the vasopressin-induced dyspeptic symptoms was mediated by central opioid peptide-producing neurons. Five female beagle dogs implanted with 1 pair of electrodes in gastric serosa were used in a two-experiment study. In experiment one, the brain was scanned by positron emission tomography in 3 dogs with and without short-pulse GES, and the radioactivity in nuclei of solitary tract (NST) and hypothalamus was detected. Experiment two was composed of 4 sessions. In session one, the dogs were injected with vasopressin in the absence of short-pulse GES. With session two, the short-pulse GES was simultaneously given via the electrodes with the injection of vasopressin. In sessions three and four, naloxone and naloxone methiodide was administered respectively in the presence of short-pulse GES. Motion sickness-like symptoms were scored and compared among the different sessions. The results showed that the short-pulse GES significantly increased the radioactivity in NST and hypothalamic nuclei (P<0.05, vs control). The short-pulse GES could ameliorate the vasopressin-induced motion sickness-like symptoms in dogs. Naloxone, but not naloxone methiodide could attenuate the curative effects of short-pulse GES. It is concluded that NST and hypothalamic nuclei may participate in the mediation of the curative effects of short-pulse GES on dyspepsia-like symptoms. Central opioid peptide-containing neurons presumably mediate the therapeutic effect on dyspeptic symptoms of short-pulse GES.

Objective: To observe the effect of Mo-Rubbing abdomen manipulation on glucose metabolism and inflammatory factors in rats with type 2 diabetes mellitus (T2DM). Methods: Sixty Sprague-Dawley rats were randomly divided into a normal group (n=10) and a group for modeling (n=50) using the random number table method. Rats in the group for modeling were induced to form T2DM models by a high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin. Thirty successfully modeled rats were randomly divided into a model group, a Mo-Rubbing abdomen group, and a metformin group, with 10 rats in each group. Rats in the normal group and the model group received no intervention, those in the Mo-Rubbing abdomen group received Mo-Rubbing abdomen manipulation, and those in the metformin group received metformin by gavage. After 8-week intervention, fasting insulin (FINS), fasting plasma glucose (FPG), homeostasis model assessment for insulin resistance (HOMA-IR) and area under the curve at the oral glucose tolerance test (AUC-OGTT), as well as serum inflammatory factors interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were detected, and the morphological changes of the pancreas were also observed. Results: After the 8-week intervention, the levels of FINS, FPG, HOMA-IR, and AUC-OGTT of rats in the Mo-Rubbing abdomen group were significantly lower than those in the model group (P<0.05); the pancreatic injury degree in the Mo-Rubbing abdomen group and the metformin group was lower than that in the model group. Compared with the model group, the serum IL-1β, IL-6, and TNF-α levels of rats in the Mo-Rubbing abdomen group were significantly decreased (P<0.05), and the serum IL-1β and TNF-α levels of the metformin group showed a downward trend; the serum IL-6 and TNF-α levels in the Mo-Rubbing abdomen group were significantly lower than those in the metformin group (P<0.01). There was a significant positive correlation between FPG with IL-1β, IL-6, and TNF-α in the T2DM rats (P<0.01). Conclusion: Mo-Rubbing abdomen manipulation reduces the inflammatory response and improves the morphological changes of the pancreas in T2DM rats, thereby achieving the effect of lowering blood glucose.

Objective:To observe factors influencing the risk of urinary tract infection (UTI) after cerebral hemorrhage (ICH).Methods:A total of 77 ICH patients undergoing rehabilitation between August 2015 and August 2017 were studied. Among them, 24 were diagnosed with a UTI according to clinical manifestations and urine routine tests and urine culture. They formed the UTI group. The other 53 were the non-UTI group. Complete clinical evaluations were available for all 77, and those data were used to identify risk factors for UTI using univariate and multivariate logistic regression analysis.Results:The univariate analysis showed that age, side of paralysis, consciousness dysfunction, Brunnstrom staging of the lower extremity, tracheotomy, indwelling catheter, lung infection, white blood cell count, neutrophil percentage, serum natrium, uric acid, D-dimer level and fibrinogen were all significant predictors of UTI. The multivariate analysis identified age, right side hemiplegia and D-dimer level as useful predictors.Conclusions:Advanced age and a high D-dimer level are independent risk factors for UTI after ICH, while hemiplegia on the right side is a protective factor.

Objective To investigate the quality of life of post-operative patients with cholecystolithiasis and analyze the influencing factors. Methods About 206 patients with cholecystolithiasis were enrolled in the investigation by gastrointestinal quality of life index (GIQLI). Multivariate linear regression analysis was used to explore the influencing factors of their quality of life. Results The GIQLI score of 206 patients was (115.81 ±9.22) and the index value was 80.42%. Three independent variables such as breakfast habit, exercise, three hyperlipidemia, were the factors that affected the quality of life (P<0.05). Conclusions The quality of life of patients after cholelithiasis is lower than that of normal people. Patients should develop good eating habits and exercise habits, strengthen the interventions with high blood lipids, high blood pressure and hyperglycemia to improve the quality of life of post-operative patients with cholecystolithiasis.

Objective To observe the changes of peripheral blood interleukin-2(IL-2) ,interleukin-4(IL-4) ,interleukin-10(IL-10) ,tumor necrosis factor-alpha(TNF-α) and interferon gamma (IFN-γ) in chronic hepatitis B(CHB) patients with positive e-anti-gen(HBeAg) treated by Bushenqingtou Decoction .Methods 60 cases of CHB with positive HBeAg were randomly divided into the treatment group and the control group ,30 cases in each group .The CHB treatment group received Bushenqingtou Decoction with the treatment course of 48 week ,while the CHB control group did not received the medication therapy .In the beginning and at 48 weeks of treatment ,the blood in the two CHB groups were collected for detecting HBV DNA ,IL-2 ,IL-4 ,IL-10 ,TNF-αand IFN-γ. Results Compared with before treatments ,the levels of IL-2 ,TNF-αand IFN-γafter treatments in the CHB treatment group were obviously increased ,while the levels of HBV DNA ,IL-4 and IL-10 were remarkably decreased (P<0 .01);however ,there were no statistical differences of the various indexes after the treatments in the CHB control group (P>0 .05) .Conclusion Bushenqingtou Decoction could inhibit the replication of HBV DNA in the CHB patients with positive HBeAg and improve the body immune func-tion .

Objective:To evaluate the efficacy of Qufeng Zhiyang Decoction combined with Fuzhiqing Ointment in the treatment of chronic perianal eczema.Methods:Randomized controlled trial. A total of 122 patients with chronic perianal eczema in Meishan People's Hospital from January 2020 to May 2021 were selected as the observation objects and divided into 2 groups according to random number table method, with 61 cases in each group. The conventional group was treated with Fuzhiqing Ointment. The study group was treated with Qufeng Zhiyang Decoction. The treatment for both groups lasted for two weeks. TCM symptom score was recored before and after treatment; the degree of skin lesions was evaluated by the area and Eczema Area and Severity Index (EASI); the quality of life of patients was evaluated by the quality of life index of dermatosis (DLQI); the number of Treg and Th17 cells was measured by flow cytometry; the ratio of Th17/Treg cells was calculated; the levels of TNF-α, IL-10 and IL-6 were detected by ELISA. The adverse reactions were recorded and the clinical efficacy was evaluated. The patients were followed up for 3 months and the recurrence was recorded.Results:The total effective rate of the study group was 86.89% (53/61) and that of the conventional group was 72.13% (44/61), with statistical significance ( χ2=4.08, P=0.044). After treatment, the scores of pruritus, spurt water, erythema and burning, upset and thirsty, yellow urine and dry urine in the study group were lower than those in the conventional group ( t values were 19.64, 49.13, 19.64, 33.14, 24.61, P<0.01); EASI and DLQI scores were lower than those in the conventional group ( t values were -6.52 and -29.33, P<0.01). After treatment, Treg cells [(6.58±0.78) % vs. (5.49±0.81) %, t=7.57] in the study group was higher than that of the conventional group ( P<0.01), Th17 cells [(1.03±0.28) % vs. (1.27±0.39) %, t=-3.90] and Th17/Treg ratio [(0.15±0.07) vs. (0.23±0.05), t=7.68] were lower than those in the conventional group ( P<0.01). After treatment, the level of IL-10 in the study group was higher than that of the conventional group ( t=-16.97, P<0.01), and the levels of IL-6 and TNF-α were lower than those in the conventional group ( t values were -21.48 and -25.59, P<0.01). During treatment, the incidence of adverse reactions was 6.56% (4/61) in the study group and 9.84% (6/61) in the conventional group, without statistical significance in the incidence of adverse reactions between the two groups ( χ2=0.44, P=0.509). During the follow-up period, the recurrence rate was 15.09% (8/53) in the study group and 50.00% (22/44) in the conventional group, with statistical significance in the recurrence rate between the two groups ( χ2=14.55, P<0.01). Conclusion:Qufeng Zhiyang Decoction combined with Fuzhiqing Ointment can reduce the levels of inflammatory factors in patients with chronic perianal eczema, maintain Th17/Treg cell immune balance, improve clinical efficacy, and have a low recurrence rate.