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Objective To study the influence of cell loss in the CA3 area on hippocampal neurogenesis in mice with temporal lobe epilepsy .Methods Newly born cells in the subgranular zone (SGZ ) of dentate gyrus were labeled by the proliferation marker bromodeoxyuridine (BrdU ) at 2 months after pilocarpine‐induced status epilepticus .Double labeling of BrdU+ NeuN was carried out to compare neuronal differentiation between type Ⅰand Ⅱ mice ,which represented partial cell loss and almost complete cell loss in the CA 3 area ,respectively .DCX and BrdU single staining were made to analyze the proliferation of progenitor cells in SGZ and the survival of the newly born cells .Results When compared with that of the type Ⅱ mice ,the number of double labeled cells of BrdU+NeuN in the subgranular zone‐granule cell layer (SGZ‐GCL) in the type Ⅰ mice increased significantly ( P<0 .05) .However ,the percentages of double labeled cells in all the BrdU + cells were found to be similar between the two groups (P>0 .05) .No significant difference in the number of DCX + cells in SGZ was found between the typeⅠ and Ⅱ mice (P>0 .05) .At 6 weeks after BrdU labeling ,more remaining BrdU+ cells were found in the SGZ‐GCL in the type Ⅰ mice when compared with their counterparts in the type Ⅱ mice (P<0 .05) .Conclusion The cell loss in the CA3 area of pilocarpine‐induced epileptic mice contributes to the declined hippocampal neurogenesis by exerting negative effects on the survival of newly born cells .

Objective To construct and identify the recombinant retroviral vector containing five copies of hypoxia responsive elements (5HRE)and neurotrophin-3 (NT-3 ).Methods Using PCR,enzyme digestion and DNA ligase,5HRE and human derived NT-3 were cloned into the retroviral vector plasmid (pLNCX)to construct the recombinant retroviral vector plasmid pLNCX-5HRE-SV40-NT3-IRES-EGFP.The retrovirus RV-5HRE-NT3 was packaged in the PT67 cells,and then it was purified and concentrated by high-speed centrifugation.After infected for 48 h with the concentrated retrovirus,the number of the EGFP positive cells in the NIH 3T3 cells was counted by fluorescence activated cells and sorted to calculate the retrovirus titer.Results The retroviral vector plasmid,pLNCX-5HRE-SV40-NT3-IRES-EGFP,was successfully constructed,and the retrovirus was packaged and defined as RV-5HRE-NT3.After purification and concentration,the retrovirus titer reached 9.1 × 10 6 cfu/mL. Conclusion The recombinant retroviral vector which carried out hypoxia-regulated expression of NT-3 was successfully constructed.It may provide basis for studies on hypoxia-regulated expression of the exogenous genes.

Objective To investigate the neuroprotective effects of neurotrophin-3 (NT-3) expression controlled by five copies of the hypoxia-responsive elements after focal cerebral ischemia .Methods Three groups of rats re-ceived RV-5H-NT3, RV-5H-EGFP or saline injection .Three days after gene transfer , the rats underwent 90 min of transient middle cerebral artery occlusion ( tMCAO) , followed by 1-28 days of reperfusion .Immunohistostaining and western blotting were performed to detect ischemia/hypoxia-regulated expression of NT-3 controlled by HRE . The volume of brain infarction and the apoptosis were analysised by TTC and TUNEL staining .The neurological scoring was determined by neurological behavior tests .Results Three days after tMCAO , brain NT-3 expression was significantly increased in the RV-5HNT3-transduced animals compared with the RV-5H-EGFP or saline group (P<0.05), and brain infarct volume was smaller in the RV-5H-NT3-transduced group than the RV-5H-EGFP or saline group ( P<0.05 ) .The percentage of TUNEL-positive cells was reduced in RV-5 H-NT3-transduced brains compared with the RV-5 HEGFP or saline group 3 and 7 days after tMCAO ( P<0.05 ) .Furthermore , the neurolog-ical status of RV-5H-NT3-transduced rats was better than that of RV-5H-EGFP-or saline-transduced animals from 1 day to 4 weeks after tMCAO ( P<0.05 ) .Conclusions HRE may modulate NT-3 expression in the ischemic brain tissue and that the up-regulated NT-3 may effectively improve neurological status following tMCAO due to de-creased initial damage .

Objective To explore the main causes for death due to severe acute pancreatitis (SAP) to improve the level of diagnosis and treatment. Methods The clinical data of 1162 SAP cases treated in our hospital from June 1997 to May 2005 were retrospectively analyzed. Among which, 144patients (12. 39%) died, 82(7.06%)abandoned treatment and 936(80.55%)were cured. Results the APACHE Ⅱ scores and pancreas Balthazar CT scores of the death group were higher than that of the survival group. The percentage of single one organ dysfunction and multiple organ dysfunction syndrome (MODS) was significantly higher in the death group than in the survival group. The mortality rate of SAP without obvious inducing factors was significantly higher than that of SAP with inducing factors. Conclusion Integrated traditional and western non-surgical treatment is effective for SAP.The treatment for SAP without obvious inducing factors is a challenge. The mortality rate of SAP is primarily related to the pathological changes of pancreas and the patient's general condition. Early diagnosis and treatment is important to decrease mortality rate and maintaining the function of important organs is basic to ensure curative effect.

Objective To explore the effect on traditional experiment and case teaching method in regional anatomy study. Methods 80 students from 2014 medical students were randomly selected as the teaching subjects and divided into traditional group and case teaching group. The traditional group con-tained 40 students, using the traditional teaching method, while case teaching group had also 40 students with case teaching method. In the process of teaching, three clinical cases were introduced, including thesubtotal thyroidectomy thoracic outlet syndrome andpancreatic cancer. After the end of the course, the students conducted a unified questionnaire and examination. SPSS 18.0 was used for data line t test or chi square test between the two groups. Results The scores of the students in the case group in the selection questions, blanks and essay questions in the final exam were higher than those of the traditional group; The average total score of the case group was (85.69 ±11.61), while the traditional group was (73.19 ±18.66), and the difference was statistically significant (t=3.597, P=0.002). The results of the questionnaire showed that the students in the case group were higher than the traditional group, and the difference was statistically significant ( χ2=14.753, P=0.001). Conclusion The effect on regional anatomy study with case teaching method is better than the traditional teaching method, and it is a promising teaching reform for the med-ical students.

OBJECTIVETo explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury.

METHODSPrimarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells.

RESULTSSalidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (P<0.05). Salidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (P<0.05).

CONCLUSIONSalidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.

Animals ; Caspase 3 ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Flow Cytometry ; Glucosides ; pharmacology ; Neural Stem Cells ; drug effects ; Phenols ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE: To investigate the relationship between behavioral changes and cell proliferation in subventricular zone (SVZ) after intracerebral hemorrhage (ICH) in adult rats. METHODS: Forty male Sprague-Dawley rats were randomly assigned into a behavioral test group (n = 19) and a bromodeoxyuridine (Brdu) immunohistochemical staining group (n = 21). ICH was induced by stereotactial injection of collagenase type VII into straitum. Proliferating cells were labeled by injection intrapenitoneally of bromodeoxyuridine in a pulse protocal. Rats were killed on day 2, 7, 14, and 28 after the ICH. Behavioral test and bromodeoxyuridine immunohistochemical staining were performed.Behavioral change was tested by forelimb placing test, Berderson's grade and corner turn test in rats. Cell counting of bromodeoxyuridine immunoreactive cells in SVZ was performed. RESULTS: There were marked neurological deficits by day 2 after the ICH, with progressive recovery of function over 4 weeks. A significant increase in the number of bromodeoxyuridine immunoreactive cells in the ipsilateral and cortralateral SVZ was observed from 2 to 14 days with a peak at day 7 after the ICH compared with the sham group.The bromodeoxyuridine immunoreactive cells decreased to control level 28 days after the ICH. CONCLUSION Proliferation of cells in SVZ corresponds well with behavioral recovery after the ICH, which indicates SVZ cells may be involved in the repairing process after the ICH.
Animals ; Behavior, Animal ; physiology ; Cell Proliferation ; Cerebral Ventricles ; pathology ; Intracranial Hemorrhages ; pathology ; physiopathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective:To explore the effects of Ophiopogon D combined with cyclooxygenase-2 (COX-2) gene silencing on the proliferation, migration and invasion of human pancreatic cancer BxPC-3 cells.Methods:BxPC-3 cells were divided into blank control group, Ophiopogonin D high-dose group (40 μmol/L), medium-dose group (20 μmol/L) and low-dose group (10 μmol/L). The COX-2-slienced cells were divided into control group, COX-2 inhibited group (50 pmol/ml siRNA-COX-2), Ophiopogonin D group (20 μmol/L) and combination treatment group (Ophiopogonin D 20 μmol/L+ 50 pmol/ml siRNA-COX-2). The proliferation activity of BxPC-3 cells was detected by CCK-8, and the migration distance of BxPC-3 cells was detected by scratched assay. The invasion degree of BxPC-3 cells was detected by Transwell, the relative expression level of COX-2 gene in BxPC-3 cells was detected by real-time quantitative PCR (RT-qPCR), and the relative expressions of COX-2, hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) proteins in BxPC-3 cells were detected by Western blotting.Results:The cell proliferation rates of blank control group, Ophiopogonin D high-dose, medium-dose and low-dose groups were (100.0±4.9)%, (71.8±5.4)%, (80.5±5.8)% and (89.7±5.7)%, respectively. The migration distances were (279.8±24.0) μm, (141.9±21.2) μm, (168.8±37.1) μm and (224.6±19.9) μm, respectively. The absorbance ( A) values of invasion number were 1.107±0.095, 0.390±0.030, 0.596±0.017 and 0.826±0.034, respectively.There were statistically significant differences ( F=19.770, P<0.001; F=48.270, P<0.001; F=198.400, P<0.001). The above indexes of the Ophiopogonin D high-, medium- and low-dose groups were significantly lower than those in the blank control group (all P<0.05). The relative expression levels of COX-2 gene were 1.007±0.178, 0.387±0.169, 0.567±0.142 and 0.740±0.030, respectively, and the relative protein expression levels were 1.000±0.033, 0.654±0.085, 0.762±0.110 and 0.881±0.049, respectively, with statistically significant differences ( F=10.280, P=0.004; F=11.780, P=0.003). The above indexes of the Ophiopogonin D high- and medium-dose groups were significantly lower than those in the blank control group (all P<0.05), and there was no statistically significant difference between the Ophiopogonin D low-dose group and blank control group (both P>0.05). The medium-dose of Ophiopogonin D (20 μmol/L) was selected as the subsequent concentration.After COX-2 silencing, the proliferation rates of the control group, COX-2 inhibited group, Ophiopogonin D group and combination treatment group were (100.0±2.8)%, (68.4±6.7)%, (67.7±5.9)% and (57.0±8.5)%, respectively, the migration distances were (274.4±23.8) μm, (217.0±18.8) μm, (186.2±18.6) μm and (115.7±15.8) μm, respectively, and the A values of invasion number were 1.143±0.092, 0.791±0.058, 0.715±0.026 and 0.424±0.058, respectively, with statistically significant differences ( F=34.430, P<0.001; F=103.400, P<0.001; F=131.100, P<0.001). The proliferation rates, migration distances and invasion numbers in each treatment group were significantly lower than those in the control group (all P<0.001). Compared with the COX-2 inhibited group and Ophiopogonin D group, the cell proliferation, migration and invasion were significantly inhibited in the combination treatment group (all P<0.05). Compared with the Ophiopogonin D group, only the migration distance of the COX-2 inhibited group was significantly different ( P<0.05). The relative expression levels of COX-2 protein in the above groups were 0.995±0.037, 0.779±0.060, 0.806±0.076 and 0.645±0.079, respectively, the relative expression levels of HIF-1α were 1.083±0.104, 0.749±0.070, 0.736±0.070 and 0.394±0.016, respectively, and the relative expression levels of VEGF protein were 1.016±0.103, 0.757±0.090, 0.745±0.021 and 0.603±0.023, respectively, with statistically significant differences ( F=14.650, P=0.001; F=45.220, P<0.001; F=18.180, P<0.001). The expression levels of the three proteins in each treatment group were significantly lower than those in the control group (all P<0.05). Compared with the COX-2 inhibited group and Ophiopogonin D group, the relative protein expression levels of COX-2, HIF-1α and VEGF in the combination treatment group were significantly decreased (all P<0.05). Compared with the Ophiopogonin D group, there were no significant differences in the expression of the three proteins in the COX-2 inhibited group (all P>0.05). Conclusion:Ophiopogon D combined with COX-2 gene silencing can inhibit the proliferation, migration and invasion of pancreatic cancer cells, and the mechanism may be related to the inhibition of COX-2 pathway and the decrease of HIF-1α and VEGF protein expression levels.

Objective: To know condom use negotiation with clients and regular sex partners and condom use in female sex workers (FSWs), and provide reference for the development of comprehensive HIV/AIDS intervention for FSWs. Methods: The cross sectional survey was conducted in Jianshui county and Mengzi county in Honghe Hani and Yi autonomous prefecture. A total of 476 FSWs aged 16 years and above were recruited from entertainment venues, and the information about their demographic characteristics, condom use negotiation and condom use were collected by using questionnaires. Logistic regression model was used to analyze related factors of condom use after negotiation between FSWs and clients unwilling use condom. Results: A total of 852 FSWs who aged (24.29±8.44) years old participated in the survey. In past month, 499 FSWs had negotiation for condom use with unwilling clients (58.6%, 499/852), after negotiation, 441 FSWs (88.4%, 441/499) had consistent condom use in each sex with the clients. In the past one month, 99 FSWs had negotiation for unwilling use condom with regular sex partners (14.4%, 99/687), after negotiation, 54 FSWs (54.5%, 54/99) had consistent condom use in each sex with regular sex partners. Among the FSWs, 266 (53.3%, 266/499) reported that they could say "It is a mandatory requirement" to persuade clients who were unwilling to use condom. 97(19.4%, 97/499) reported that they could say "There is risk for infection" to persuade clients who were unwilling to use condoms. 115 (23.1%,115/499) reported that they could say "It is a mandatory requirement" and "there is risk for infection" to persuade their unwilling clients to use condoms. 21 (4.2%, 21/499) reported that they used other strategies. 22 (4.4%, 22/499) felt that it was difficult to persuade clients to use condoms. Multivariate logistic regression analysis showed that compared with FSWs who felt difficult in persuading clients to use condoms, FSWs who felt moderate difficulty were more likely to have consistent condom use after negotiation (OR=4.00, 95%CI: 1.55-10.32) and FSWs who felt easy in persuading clients to use condoms were also more likely to have consistent condom use (OR=30.17, 95%CI: 3.22-282.44). Compared with FSWs used other strategies to persuade their clients to use condoms, FSWs who said it was a mandatory requirement were more likely to have consistent condom use after negotiation (OR=4.44, 95%CI: 1.41-14.01) and FSWs who said it was a mandatory requirement and there was risk for infection were also more likely to have consistent condom use (OR=5.52, 95%CI: 1.55-19.73). Conclusions Negotiation for condom use increased the rate of condom use in FSWs in sex with clients who were unwilling to use condom. The negotiation strategy of "It is a mandatory requirement" would promote condom use in FSWs in sex with clients who were unwilling to use condom. Besides, the negotiation strategy of saying "there is risk for infection" had additional effects.