Practice and exploration of University of Science and Technology of China in improving efficiency during formality examination of the National Natural Science Fund Project / 中华医学科研管理杂志

Objective To improve the efficiency and accuracy of formality examination of the National Natural Science Fund Project (hereinafter referred to as formality examination).Methods According to the practice,we find that Combine Files of Adobe Acrobat Pro software,which relies on the large data of internet-based Science information system (hereinafter referred to as the system) can quickly search and examine the various problems in the application.Results In previous years,we examined the PDF version or hard copy of the application form of NSFC according to the list of questions about formality examination.It is easy for the examiner to miss the examination due to fatigue after long time working which resulted in many amendments by the applicant.Relying on the above two technologies,we redesigned and constructed a three-level formality examination process,reduced the burdens of examiner,and also greatly improved the efficiency and accuracy of formality examination.Conclusions Finding and using the right tools can lead to amazing productivity gains in boring management.

Effects of quinolinic acid on autophagy and protein expressions of related signaling pathway in PC12 cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate whether quinolinic acid(QA)induces autophagy in PC12 cells and its relationship with glycogen synthase kinase-3β(GSK-3β)/β-catenin related signaling path?ways. METHODS PC12 cells were treated with QA 2.5,5.0 and 10.0 mmol·L-1 for 24 h. The cell viability was determined by MTT assay. Autophagy fluorescent spots labelled form of microtubule-associated protein 1 light chain 3(LC3)was examined by LC3 immunostaining. The expressions of GSK-3β,β-catenin,LC3 and Beclin 1 were determined by Western blotting. RESULTS QA inhibited PC12 cell survival in a concentration-dependent manner,and IC50 was 8.7 mmol · L- 1. Compared with normal control group,QA 2.5,5.0 and 10.0 mmol · L-1 increased autophagic intracellular LC3 fluorescence spots,elevated the expression ratio of LC3-Ⅱ/LC3-Ⅰ and expression of Beclin 1 in PC12 cells(P<0.05). In addition,QA enhanced GSK-3βexpression and decreasedβ-catenin expression(P<0.05,P<0.01). CONCLUSION QA induces autophagy in PC12 cells. This mechanism may be associated with the activation of GSK-3β/β-catenin related signaling pathways.

Construction of human telomerase reverse transcriptase-immortalized rat bone marrow mesenchemal stem cell strains / 中国组织工程研究

BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.

Effects of nutrient fortified complementary food supplements on anemia of infants and young children in poor rural of Gansu / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To assess the effectiveness of complementary food supplements with protein and multi-micronutrients on hemoglobin and anemia in infants and young children.</p><p><b>METHODS</b>In 5 poor counties of Gansu, 984 children aged 6-12 months were enrolled and divided into two groups. In addition to the usual home-made complementary food, all the children were fed one sachet of either Formula I or Formula II supplements each day. Protein and micronutrients were provided in Formula I, while the same energy intake was secured in Formula II as in Formula I. A massive dose of vitamin A was supplemented to all the children every 6 months. Hemoglobin test was done at the same time.</p><p><b>RESULTS</b>Prevalence of anemia was about 35% in both Formula I and Formula II group at baseline, and there were no differences in hemoglobin concentration between the two groups. During the 6-month and 12-month supplementation, hemoglobin of children in Formula I group was higher than that in Formula II group (P < 0.05), and hemoglobin increase in Formula I group was significantly higher than that in Formula II group (P < 0.001). After 6- and 12-month supplementation, the prevalence of anemia in Formula I group dropped to 19.1% and 8.2% respectively, and it was 28.0% and 12.4% in Formula 2 group. The prevalence of anemia in Formula I group was significantly lower than that in Formula II group (P < 0.05). After adjusting age and hemoglobin level at baseline, the hemoglobin increase at age of 24 months in formula 1 group was higher (10.7 g/L vs 7.9 g/L, P < 0.0001).</p><p><b>CONCLUSION</b>Micronutrient fortified complementary food supplements, with large-dose vitamin A, is effective for children aged 6-12 months in terms of iron deficiency prevention.</p>

Evaluation of NR2B peptide as subunit vaccines against experimental neuropathic pain / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>NR2B containing N-methyl-D-aspartate (NMDA) receptor plays an important role in the facilitation and maintenance of neuropathic pain. The discrete distribution of NR2B subunit in the central nervous system (CNS) may support reduced side effects of agents that act selectively at this site. Therefore, we investigated the hypothesis that a humoral autoimmune response targeting the NR2B subunit of NMDA receptor relieves pain like behaviours produced by peripheral injury.</p><p><b>METHODS</b>Rats were immunized subcutaneously with NR2B-Keyhole Limpet Hemocyanin (NR2B-KLH) three times at two-week intervals. NR2B specific IgG titres in sera and cerebrospinal fluid were determined by indirect ELISA. Seven days after the third immunization, 2 of the 3 terminal branches of the sciatic nerve (tibial and common peroneal nerves) were tightly ligated. Behavioural testing was carried out on every other day after surgery, until 7 days after surgery. The lumbar spinal cord (L4-6) was removed on day 7 after ligation. The expression of NR2B protein in the lumbar spinal cord was determined using Western blotting.</p><p><b>RESULTS</b>After the second vaccination, NR2B specific IgG in sera was detected to be > 0.5 microg/ml in six of nine rats. After the third vaccination, all the immunized rats had > 2.2 microg/ml. Titres of NR2B specific IgG in sera peaked 42 days post initial immunization and persisted for over 70 days. No NR2B specific IgG was detected in sera from PBS or KLH group. The behavioural thresholds in NR2B group were significantly higher than those in PBS and KLH groups on day 7 after ligation. NR2B specific IgG in CSF in NR2B group could not be detected on day 1 before spinal dissection; but could be detected on day 7 after surgery. The expression of NR2B protein in group NR2B was significantly lower than in PBS and KLH groups on day 7 after surgery.</p><p><b>CONCLUSION</b>The NR2B peptide could be used as a vaccine against neuropathic pain, which could be associated with reduction of NR2B protein in the lumbar spinal cord.</p>

Construction and identification of immortalized rat astrocyte cell line expressing enkephalin / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To provide a sound cell source for further ex-vivo gene therapy for chronic pain, we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline.</p><p><b>METHODS</b>Retrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline. The hPPE gene expression level of immoralized astroyte cells (IAC)/ hPPE was detected by RT-PCR, indirect immunofluorescence staining and radioimmunoassay.</p><p><b>RESULTS</b>IAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline.</p><p><b>CONCLUSION</b>An immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully, which provides a research basis for transgenic cell transplantation for analgesia.</p>

Immortalized rat astrocyte strain genetically modified by rat preprogalanin gene / 华中科技大学学报（医学）（英德文版）

To construct an immortalized rat astrocyte strain genetically modified by rat preprogalanin gene (IAST/GAL) and detect its galanin (GAL) expression and secretion, a cDNA fragment of rat GAL in plasmid of pBS KS(+)-GAL was inserted into eukaryotic expression vector pcDNA3.1 (+) by DNA recombinant technology, then the restriction enzyme digestion and DNA sequencing were carried out to evaluate the recombinant. The pcDNA3.1 (+)-GAL and pcDNA3.1 (+) construct were transfected into immortalized rat astrocyte strain (IAST) by lipofectamine and the population of cells which stably integrated the construct was selected with 600 microg/mL G418. Individual clones were screened and expanded into clonal cell strains. Detection of Neo gene was used to validate the success of the transfection. Immunocytochemical staining, RT-PCR and radioimmunoassay were used to detect the expression and secretion level of GAL. The recombinant had been successfully constructed by restriction enzyme digestion and DNA sequencing. Detection of Neo gene showed that the pcDNA3.1 (+)-GAL and pcDNA3.1 (+) have been successfully transfected into IAST. After selection by using G418, IAST/GAL and IAST/Neo cell strains were obtained. IAST/GAL, IAST/Neo and IAST were immunostained positively for GAL, but the GAL average optical density of IAST/GAL was significantly higher than that of IAST/Neo and IAST (P0.05). It was concluded that IAST/GAL strain was constructed successfully and it might provide a basis for the further study of pain therapy.

Immortalized Rat Astrocyte Strain Genetically Modified by Rat Preprogalanin Gene / 华中科技大学学报（医学）（英德文版）

To construct an immortalized rat astrocyte strain genetically modified by rat preprogalanin gene (IAST/GAL) and detect its galanin (GAL) expression and secretion, a cDNA fragment of rat GAL in plasmid of pBS KS(+)-GAL was inserted into eukaryotic expression vector pcDNA3.1(+) by DNA recombinant technology, then the restriction enzyme digestion and DNA sequencing were carried out to evaluate the recombinant. The pcDNA3.1 (+)-GAL and pcDNA3.1 (+) construct were transfected into immortalized rat astrocyte strain (IAST) by lipofectamine and the population of cells which stably integrated the construct was selected with 600 μg/mL G418. Individual clones were screened and expanded into clonal cell strains. Detection of Neo gene was used to validate the success of the transfection. Immunocytochemical staining, RT-PCR and radioimmunoassay were used to detect the expression and secretion level of GAL. The recombinant had been successfully constructed by restriction enzyme digestion and DNA sequencing. Detection of Neo gene showed that the pcDNA3.1 (+)-GAL and pcDNA3.1 (+) have been successfully transfected into IAST. After selection by using G418, IAST/GAL and IAST/Neo cell strains were obtained.IAST/GAL, IAST/Neo and IAST were immunostained positively for GAL, but the GAL average optical density of IAST/GAL was significantly higher than that of IAST/Neo and IAST (P＜0.01). The level of GAL mRNA expression and the supernatant concentration of GAL in cultured IAST/GAL were significantly higher than those of IAST and IAST/Neo (P＜0.01), but no significant differences were found between the IAST and IAST/Neo (P＞0.05). It was concluded that IAST/GAL strain was constructed successfully and it might provide a basis for the further study of pain therapy.

The study on relationship of body mass index and blood pressure in children and adolescents of Beijing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To identify the relationship of body mass index (BMI) and blood pressure in 7 - 15 years children and adolescents of Beijing so as to provide scientific basis for early prevention of hypertension and to provide evidence for verification on the category criterion of overweight and obesity in children and adolescents of China, recommended by the Working Group on Obesity in China (WGOC) to sensitively distinguish the blood pressure in normal weight, overweight and obesity populations.</p><p><b>METHODS</b>A cross-sectional survey on epidemiological characteristics of obesity with stratified cluster sampling method carried out in Beijing in April and May, 2000. 5155 students aged 6 - 15 years were selecte das research subjects. The category criterion of overweight and obesity in children and adolescents of China was recommended by WGOC, the diagnostic criterion of hypertension in children was recommended by CDC in the USA. Statistics analysis system (SAS 8.1) including partial person correlation analysis, t-test, chi(2) test and logistic multi-factors regression analysis was used to analyses the data from 4982 subjects aged 7 - 15 years.</p><p><b>RESULTS</b>(1) after the age and gender were adjusted, the BMI positive correlation with systolic blood pressure (SBP) and diastolic blood pressure (DBP) was found independent in 7 - 15 years children and adolescents (P < 0.0001) and the partial relation coefficients(r) between BMI and SBP and DBP were 0.323 87 and 0.245 88 respectively. (2) the means of SBP and DBP in obesity group were significantly higher then overweight, while overweight was significantly higher then normal weight group (P < 0.0001). (3) the prevalence rates of hyper-SBP, hyper-DBP and hypertension were significantly different (P < 0.0001). When compared with the normal weight group, the relation risk (RR) for hypertension in overweight group and obesity group were 2.96 and 4.85 respectively. The prevalence rates of hypertension in overweight and obesity group were 19.70% and 24.22% respectively. (4) the results of logistic multi-factors regression analysis showed that both age and weight were effecting on hyper-SBP, hyper-DBP and hypertension (P < 0.0001). After age was adjusted, the RR for hypertension was 2.62, and their confidence interval (CI) was 2.36 - 2.91 in obesity or overweight, between overweight and normal weight.</p><p><b>CONCLUSION</b>(1) the BMI positive correlation with SBP and DBP was found independent in 7 - 15 years children and adolescents of Beijing, and the risk for hypertension maybe increased when these people with overweight and obesity, it is very important for hypertension prevention and control that overweight and obesity prevention and control in children and adolescents. (2) the sensitivity of the category criterion of overweight and obesity in children and adolescents of China, recommended by WGOC have been verified on distinguish the blood pressure in normal weight, overweight and obesity populations.</p>

Study on the utilization of health services and costs of hospital-based medical care for 29 patients with HIV/AIDS in China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To describe the costs of medical care and utilization of health service of patients with HIV/AIDS.</p><p><b>METHODS</b>Patients with human immunodeficiency virus (HIV)/AIDS treated in the Beijing You'an Hospital were interviewed retrospectively during December 1999. Data on demographic characteristics, disease process, and utilization of health service and costs of hospital-based medical care were collected.</p><p><b>RESULTS</b>A total number of 29 patients with HIV/AIDS were interviewed, including 17 (58.62%) asymptomatic HIV infections and 12 AIDS patients. Asymptomatic HIV infections had a mean of 6 outpatient visits, 1.3 hospitalizations and 58.6 inpatient hospital days per person-year. AIDS patients made, on average, 7.8 outpatient visits, 2.1 hospitalizations and 200.2 inpatient hospital days per person-year. The outpatient and inpatient medical costs were 13,729 RMB and 4,745 RMB for asymptomatic HIV infections, and 15,053 RMB and 22,242 RMB for AIDS patients per person-year respectively. For those who took both outpatient and inpatient medical care, the medical care costs, including costs of outpatient care and those of inpatient care, were 16,248 RMB for asymptomatic HIV infections and 36,795 RMB for AIDS patients.</p><p><b>CONCLUSION</b>Demands for health services and costs for medical care were high among patients with HIV/AIDS. Further study on utilization of health services and cost of medical care for patient with HIV/AIDS in a wider geographic coverage are needed.</p>

Construction of immortalized rat astrocyte strain by transfection of simian virus 40 large T antigen gene / 中华麻醉学杂志

Objective To immortalize rat astrocytes which could be used as cell carriers for transgenic cellular analgesia. Methods Astrocytes were isolated from cerebral cortex of newborn SD rats by trypsin digestion and cultured according to the method of differential cell adhesiveness and transfected with plasmid pCMVSV40T/ PUR containing the simian virus 40 large tumor antigen (SV40Tag) gene. The positive colonies were isolated by puromycin selection and expanded by many passages. The integration and expression of large T antigen gene were detected and the immuno-reactivity of glial fibrillary acidic protein (GFAP) was determined by PCR, RT-PCR and immuno-cytochemistry. Results Rat astrocytes were successfully cultured in vitro and positively stained for the astrocytic marker GFAP. The positive colonies were isolated and subcultured for 50 passages. PCR and RT-PCR products were analyzed using 1.5% agarose gel electrophoresis. The size of amplifacation product of target gene was identical to that of the positive control (558 bp) . There was no PCR and RT-PCR product from non-transfected cells. DNA sequencing and BLAST showed that 558 bp nucleotides were identical to the SV40Tag gene of the Genbank (100%). The transfected cells were positively stained for the SV40Tag and GFAP. Conclusion Immortalized rat astrocyte strain with SV40 tag gene is constructed successfully.

The role of neuronal and glial activation in spinal cord in the mechanism of neuropathic pain following three different peripheral nerve injuries in rats / 中华麻醉学杂志

Objective To determine if there is any difference in neuronal and glial(astrocytic and microglial)activation in the spinal cord in three rat models of neuropathic pain.Methods Twenty-four SD rats weighing 150-200 g were randomly divided into 4 groups(n=6 each):Ⅰ control group;Ⅱ chronic constrictive injury group(CCI);Ⅲ spinal nerve ligation group(SNL)and Ⅳ spared nerve injury(SNI).No operation was performed in control group.In CCI group left sciatic nerve was exposed and loosely ligated with catgut.In SNL group the L_5 spinal nerve was exposed and ligated with silk suture and cut.In SNI group tibial nerve and common fibular nerve were ligated and cut.Pain threshold was measured using plantar tactile stimulator(Ugo,Basile Co. Italy)every other day from 3 days before until 15 days after operation.50% paw withdrawal threshold was measured using up-and-down sequential mechanical stimulation of different intensity(0.45,0.70,1.20,2.00, 3.63,5.50,8.50,15.10 g)applied to the plantar surface of the injured paw.On the 15~(th) day after operation after pain threshold was measured the animals were anesthetized with intraperitoneal 10% chloral hydrate 400 mg? kg~(-1).The L_(5,6) segment of the spinal cord was isolated.Neuronal,astrocytic and microglial activation was determined by immuno-histochemistry with antibodies of c-Fos(a proto-oncogene protein),GFAP(an astrocyte marker)and OX-42(a microglial marker).Results The 50% paw withdrawal threshold reached the lowest level on the 7~(th) day after operation.The lowest level was maintained until the 15~(th) day after operation in group CCI,SNL and SNI.The 50 % paw withdrawal threshold was(14.1+1.5)g in control group,(2.5+0.5)g in CCI group, (1.5?0.6)g in group SNL and(0.8?0.4)g in group SNI.The number of c-Fos positive neurons in laminae Ⅳ-Ⅵ of dorsal horn was significantly greater in group CCI,SNL and SNI than in control group,but there was no significant difference among the 3 peripheral nerve injury groups.The activation of astrocytes and microglias in laminae Ⅰ-Ⅳ of dorsal horn was significantly increased in group CCI,SNL and SNI than in control group but there was no significant difference among the 3 peripheral nerve injury groups.Condusion There is no significant difference in activation of neurons and astrocytes and microglias in the ipsilateral dorsal horn among the 3 pain models.

Biological character of immortalized rat astrocyte strain by transferction of simian large T antigen gents / 中华麻醉学杂志

Objective To study the biological character of rat astrocyte strain immortalized by simian virus 40 large T antigen gene(SV40Tag) and explore the feasibility of using it as cell vehicle for transgenic cellular analgesia.Methods Rat cerebral cortical astrocytes (AST) and immortalized rat astrocyte strain (IAST) were cultured in vitro. Morphology and growth features of AST and IAST were examined and compared after subculture, freezing and recovery. The ultrastructure of the cells was observed by transmission electron microscopy. The glial fibrillary acidic protein (GFAP) in these cells was detected by immuno-cytochemistry. The cell proliferation rate and cell cycle were determined by bromodeoxyuridine labelling and flow cytometry. AST and IAST were cultured in soft agar and inoculated in nude mice to investigate the tumorigenesis of IAST. Results LAST could be subcultured successively. The cells remained monolayer, anchorage dependent and the growth was attachment-inhibited. When AST was subcultured for only 10 passages, replicative senescense began. Subculture, freezing and recovery did not influence the shape and proliferation of IAST (94%?5%) , but decreased the vitality rate of AST (54%?4% ) (P

Determination of Estrogens in Water by Solid Phase Extraction-High Performance Liquid Chromatography / 环境与健康杂志

0.998. The detection limits were in the range of 16-40 ng/L(S/N=3). The quantitative detection limits were in the range of 45-105 ng/L(S/N=10) with recovery percentages ranging from 87.6% to 106% and the relative standard deviations was lower than 3%. Conclusion The method was proved to be satisfactory in precision,accuracy and sensitivity and applicable to the daily test of estrogens in water.