Aim To investigate the effects of chloride channel blockers on the apoptosis of RIN-m? cells of pancreatic islet induced by H2O2.Methods The apoptotic model was made by H2O2 exposed for six hours with a concentration of 500 ?mol?L-1.The chloride channel blockers:DIDS,NPPB and NFA were administered to pretreat the samples respectively.The cell viability,morphological changes,and apoptosis rate were observed.Results Chloride channel blockers alone have no marked effects on the cell viability of RIN-m? cell.However,they elevated the cell viability of RIN-m?cell disposed of by H2O2.Compared to H2O2 group,the groups of DIDS +H2O2,NPPB+ H2O2 and NFA+H2O2 have significant difference in cell viability and apoptosis rate(P

The influence of Rosa davurica pall juice on pulmonary adenoma induced by urethan was observed in the experiment, which lasted for 20 weeks. 250 mice were divided into 4 groups at random. Group A was set as control receiving basal diet and tap water only. Another 3 groups were given basal diet, however, group B was given Rosa davurica pall juice 0.2ml/20g boody weight once a week, group C 0.1% urethan solution for drinking freely and group D 0.1% urethan solution plus the juice. The mouss body weight was weighed once a week; the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), the level of lipoperoxides (LPO), lymphocyte transformation and hemolysin were determined once a month, the pulmonary pathological examination was made also once a month. The obtained data were carried out with a 2?2 factorial analysis. The result were as follows:(1)the incidence of pulmonary adenoma of group C(100%) was significantly higher than that of group D (69%) at the e-nd of experiment (P

A rapid analysis method of piperazine ferulate tablets by optic-fiber sensing technology with UV-vis absorption spectrum was established.Qualitative and quantitative data were obtained and compared by maximum and minimum wavelength,absorbance and contrast spectra.Similarity method was used to identify authenticity of drugs.The difference of contents measured by this method and UV determination method in China Pharmacopoeia showed no statistical significance (P＞0.05),while the similarity can be used as a parameter to identify the authenticity of drugs.

BACKGROUND: MyoD gene is one of family members of muscle transcription factors. Transfection MyoD gene can switch on the procedure of differentiation of muscles, and transit non-muscle cells into muscle cells.The MyoD gene only expresses in skeletal muscles. Based on the same contractive structure in myocardial cells and skeletal muscle cells, it is imagined that the conversion from exogenous MyoD gene-induced fibroblast in local myocardium into skeletal muscle cells that had contractive function may become another method in the treatment of congestive heart failure on clinic.OBJECTIVE: To construct and identify a recombinant adenoviral vector carrying MyoD gene for further studies on the recovery function of MyoD gene in myocardial injury.DESIGN: Single sample experiment.SETTING: Department of Cardiology, First Affiliated Hospital, Nanjing Medical University.MATERIALS: The experiment was conducted at the Laboratory of Microbiology and Immunology, Nanjing Medical University between September 2004 and September 2005. MyoD gene and non-replicating form expressive vector of adenovirus were taken as research materials.METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction (PCR), and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.MAIN OUTCOME MEASURES: Evaluation of PCR and DNA sequencing were used for confirming the size of segment and correctness of rank of MyoD cDNA and detecting the titre of virus.RESULTS: MyoD recombinant adenovirus contained target segment with precise length confirmed by PCR and DNA sequence that was correct. The titre of virus was 1.3×1011 pfu/mL.CONCLUSION: The recombinant adenoviral vector carrying MyoD gene is constructed successfully.

Forensic toxicology is a specialized subject which forensic medical appraisers and students must learn and master. As a new teaching mode,problem based learning method is attrcting a lot of attention. Applying "case" based PBL teaching method is an important method to train innovative and high technical ability forensic talents.

Objective To compare the effects of isoflurane or sevoflurane in combination with remifentanil anesthesia on blood amyloid beta protein (Aβ) in the elderly patients undergoing abdominal surgery.Methods Two hundred patients of both sexes,aged 65-75 yr,weighing 51-76 kg,of ASA physical status Ⅰ or Ⅱ,scheduled for elective abdominal surgery under general anesthesia,were randomly divided into 2 groups (n =100 each) using a random number table:isoflurane combined with remifentanil anesthesia group (IR group) and sevoflurane combined with remifentanil anesthesia group (SR group).Fifty healthy elderly subjects served as control group (group C).After anesthesia was induced with iv penehyclidine,sufentanil,propofol and vecuronium,the patients were endotracheally intubated and mechanically ventilated.In group IR,anesthesia was maintained with inhalation of isoflurane (end-tidal concentration 1.68 ％,in IR group) or sevoflurane (end-tidal concentration 1.71％,in SR group),and target-controlled infusion of remifentanil (target plasma concentration 2-6 ng/ml).At l day before surgery and 3 days after surgery,the patients' cognitive function was assessed using Mini-Mental State Examination (MMSE),the development of postoperative cognitive dysfunction (POCD) was recorded,and blood samples were taken for determination of serum Aβ40 and Aβ42 concentrations.Results The incidence of POCD was 5％ (in C group),56％ (in IR group) or 22％ (in SR group),and there was no significant difference among the three groups.There were no significant differences in the serum Aβ42 and Aβ40 concentrations after surgery among the three groups.Conclusion The mechanism by which sevoflurane or isoflurane in combination with remifentanil anesthesia results in POCD is not related to the levels of blood Aβ40 or Aβ42 in the elderly patients undergoing abdominal surgery.

Objective:To prepare anti-human c-kit monoclonal antibody(McAb) by genetic immunization in spleen,and to determine practicability of these means to produce McAbs based on the biological activity of anti-human c-kit antibody.Methods:Recombinant plasmid pcDNA3.1/c-kit extracellular domain was constructed by molecular cloning techniques,and was used to immunize BALB/c mice in spleen directly to prepare mAb against human c-kit by routine hybridoma technique.FASC、fluorescence microscope and Western blot were utilized to identify the prepared antibody.Results:c-kit extracellular region was cloned and insert pcDNA3.1 plasmid successfully.Three hybridoma cell lines 6C4、2C5 and 5D5 that secrete anti-human c-kit McAbs were obtained after using intra-spleen immunization with a DNA vaccine.The isotypes of these three antibodies were all IgM,and the epitopes were different with each other.Conclusion:The method of genetic immunization into spleen can be used to prepare anti-human c-kit monoclonal antibodies.

BACKGROUND:Establishing a characteristic.stable and repeatable model of Th1/Th2 imbalance in animals,is the key of studying the mechanism of Th1/Th2 imbalance.OBJECTIVE:To observe the characteristics of Th1/Th2 imbalance in splenocytes derived from Balb/C mice immnnized by keyhole limpet hemocyanin(KLH).DESIGN:A randomized control exploratory experiment.SETTING:Hebei Provincial Forensic Laboratory.Institute of Basic Medicine,Hebei Medical University.MATERLALS:The experiment was carried out in the Hebei Provincial Forensic Laboratory,Institute of Basic Medicine,Hebei Medical University from September 2005 to January 2007.Balb/C mice were adopted in this study.and all the disposals were in accordance with the guidance of animal ethics.METHODS:Balb/C mice were immunized with KLH emulsified in complete Freund's adjuvant(CFA),splenocytes were acquired,and the peak of cytokine secretion was determined in 3 groups:KLH groups of 6.25 mg,kg.12.5 mg,kg and 25 mg/kg.According to the immunizing dose and immunizing frequency.mice were divided into 7 groups:KLH groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,secondary immunity groups of 6.25 mg/kg,12.5 mg/kg and 25 mg/kg,as well as control group.According to the determined levels of IgG1 and IgG2a in blood serum.mice were divided into KLH group of 6.25 mg/kg and control group.MAIN OUTCOME MEASURES:Mice splenocytes supematant was detected with enzyme linked immunosorbent assay (ELISA)for the production of Th1 cytokines interferon (IFN)-γ,interleukin(IL)-2.IL-12 p40 and Th2 cytokines IL-4 and IL-5.The levels of Th1 antibody IgG2a and Th2 antibody IgG1 in blood serum were also detected by ELISA.RESULTS:The spleen derived from KLH-immunized mice appeared hypertrophy,and the number of splenocytes was manifold.Splenocytes restimulated with KLH in vitro produced much more IFN-γ,IL-2,IL-4,IL-5,but not IL-12p40.IL-2 secretion was obviously elevated after incubated for 24 hours and achieved pinnacle at 48 hours;productions of IL-4,IL-5 and IFN-γ were elevated after 24 hours,and increased gradually to 96 hours;IL-12p40 production was very low at every time point.Using different doses of KLH inlmunity once or twice,could all lead to the elevated productions of IL-2,IL-4,IL-5,IFN-γ,and the elevation of IL-4/IFN-γ ratio,but the secondary immunity group of 6.25 mg/kg KLH showed obviously higher levels than other groups(P＜0.01).The level of KLH specific antibody IgG2a and IgG1,especially IgG1 was elevated in blood serum of KLH-immunized mice.CONCLUSION:Balb/C mice immunizad with KLH emulsified in CFA can indce a Th2 predominant imbalance in splenocytes.

AIM:To observe the effects of cholecystokinin octapeptide (CCK-8) and its receptor antagonists on cAMP response element binding protein ( CREB) and phosphorylated CREB ( pCREB) expression in frontal cortex and hippocampus of morphine withdrawal rats , which aim to explore the post-receptor mechanism through which CCK-8 regu-lates morphine withdrawal .METHODS: After the morphine dependence and naloxone-precipitated withdrawal animal models were established, the effects of CCK-8, L-364718 (CCK1 receptor antagonist) and LY-288513 (CCK2 receptor an-tagonist) pretreatment on CREB and pCREB expression in frontal cortex and hippocampus were observed by Western blot -ting and immunohistochemistry .RESULTS:In rat frontal cortex neuron , CREB was expressed in both cytoplasm and nu-cleus, but pCREB was only highly expressed in the nucleus .In the pyramidal cell layer of hippocampal CA 1 region, CREB showed high expression in the cytoplasm and low expression in the nucleus , while pCREB was only expressed in the nu-cleus.No obvious change of CREB was observed after either chronic morphine treatment or naloxone withdrawal .The pCREB expression was increased after chronic morphine treatment and further increased after naloxone withdrawal .Com-pared with the withdrawal group , chronic pretreatment with CCK-8, L-364718 and LY-288513 had no effect on CREB expression in the frontal cortex , but obviously decreased the pCREB expression .In the hippocampus , pretreatment with L-364718 and LY-288513 decreased CREB and pCREB expression , but only the pCREB expression was decreased after CCK-8 treatment.CONCLUSION:CCK-8 and CCK receptor antagonists may alleviate morphine withdrawal symptoms by regulating CREB , with specificity in different brain regions .

Aim To study targeting capability of anti-CD19 (Fab)-LDMto CD19 +B lymphoma cells in vi-vo and in vitro.Methods Flow cytometry was em-ployed to determine the affinity of Cy5 labeled anti-CD19 (Fab)-LDP to human lymphoma Raji cells.And the optical imaging system was used to analyze the dis-tribution of Cy5-anti-CD19 (Fab )-LDP in lymphoma-transplanted xenograft nude mice in vivo.Results The results of flow cytometry demonstrated that Cy5-an-ti-CD19(Fab)-LDP had remarkable affinity with lym-phoma Raji cells;Raji lymphoma xenograft model was established successfully in nude mice and in vivo fluo-rescence imaging analysis indicated that the antibody-drug conjugates could specially be localized in the tar-get tumor.Conclusion The experiments in vivo and vitro confirm that anti-CD19 (Fab)-LDP has remarka-ble affinity to targeting CD19 +lymphoma cells,and the antibody drugs anti-CD19 (Fab )-LDP have the probability to be new drugs for the treatment of malig-nant lymphoma.