Through intensifying the clinical pathological discussions and clinical pathological dissections,the clinical thinking,iatri- cal responsibility and analytical ability of the medical students have been improved markedly and the abridge function of pathology between basic medicine and clinical medicine has also been reinforced.

[Objective] To observe the clinical efficacy of ZICAOYOUSHA in treating diabetic foot ulcers.[Method] A multi-center ,randomized,double-blind,placebo-control ed study was conducted. A total of 232 patients with diabetic foot ulcers were randomly assigned the treatment group and control group,in foundation treatment at the same time,the therapy group which was treated by External Application ZICAOYOUSHA had 174 patients,the contrast group which was treated by External Application Gentamicin Emery cloth had 58 patients. Observe the aspect improvement situation in two groups separately in accordance with Wagner grading,carry out statistics processing.[Results] Two groups of curative effect indices had significant differ-ence. [Conclusion] ZICAOYOUSHA is an effective drug for external use in treating diabetic foot ulcers.

Objective To compare the analgesia and side-effects of different concentrations of sufentanil combined with 0.125% ropivacaine for postoperative epidural analgesia after general thoracic surgery.Methods Thirty-six ASA Ⅰ-Ⅲ patients (25 males, 11 females) ages 21-64 yrs weighing 42-79 kg undergoing elective general thoracic surgery under general anesthesia were randomly assigned to receive patient-controlled epidural analgesia (PCEA) with 0.125% ropivacaine combined with sufentanil 0.4 (group A, n = 12) , 0.5 (group B, n = 12) or 0.6 ?g?ml-1 (group C, n = 12) . Epidural catheter was placed at T7,8 or T8,9 interspace. The PCEA pump was set up with back ground infusion of 2 ml?h-1 , a3ml bolus dose and a 30-min lock-out period. VAS scores was used to assess analgesia at rest and during movement. The total bolus doses, PCEA button pressing times (effective/actual), vital signs including MAP, HR, respiratory rate, SpO2 and side effects (nausea, vomiting, pruritus and dyspnea) were recorded. Results During the 48 hours after operation the VAS scores in group C were significantly lower than those in group A and B ( P

Objective To employ 2 approaches to construct and express reconstructed LL-37 (rLL-37) in procaryotic system, and to explore a better preparation method. Methods The first method: the rLL-37 was inserted into vector pET-28a (+), then was induced to express in E.coli. BL21 (DE3) and purified by chromatography; the second method: the rare codons in the rLL-37 gene sequence were substituted by the preferred codons of procaryotic cell, and a fragment of carrier protein molecule (CPM) was added to the N termination of the objective sequence to construct expression plasmid pET-30a(+)-CPM-rLL-37, then the rLL-37 was expressed in E.coli. BL21 Star(DE3) and purified by chromatography. The productive rates of the 2 methods were compared and the antimicrobial effects of obtained rLL-37 was studied. Results The first method: the DNA sequence of rLL-37 was obtained successively by Touch-Down PCR. The expression plasmid pET-30a(+)-CPM-rLL-37 was expressed with fusion protein in E.coli BL21 (DE3). The expression rate accounted for 20% of total bacterio-protein, then the expressed product was purified by using high positive ion exchange column Macro-Prep High S; The second method: a fragment of carrier protein molecule was designed that contained 28 amino-acid residue and its pHi was 2.7, net charge was-6.0 at pH 7.4. After the expression plasmid pET-30a(+)-CPM-rLL-37 was constructed successively, it was expressed in E.coli BL21 Star (DE3). The expressed fusion protein accounted for 35% of total bacterio-protein, then the expressed product was purified by using affinity binding chromatography with TALON resins successfully. The obtained 2 kinds of rLL-37 were able to kill both Gram-negative and-positive bacteria by the means of inhibitory zone. Conclusion It’s feasible to prepare efficiently rLL-37 in procaryotic system, which founds the basis for the further research on bactericidal activity of rLL-37.

Objective To obtain the disulfide stabilized Fv fragment (dsFv) against N-terminal fragment of human lip polysaccharide binding protein (NH-LBP) and to identify its biological vitality. Methods The disulfide stabilized Fv fragment antibody (dsFv) was obtained after the inclusion bodies of dsFvVH and dsFvVL had been refolded and purified. Then the characteristics of dsFv were determined in vitro by ELISA and by detecting the secretion of tumor necrosis factor alpha (TNF-?) in rats. Results There was 2.1 mg protein of dsFv obtained. dsFv had good combination with NH-LBP and could restrain inflammatory reaction caused by lip polysaccharide (LPS) in vivo. Conclusion It is feasible to get dsFv against NH-LBP by respective expression of VL and VH. The partial inhibition of the biological function of LBP by dsFv is a new way to restrain the over-inflammatory reaction in vivo.

Objective To investigate the clinical therapeutic effect of acupuncture on pulmonary infection after acute cerebral infarction.Methods Seventy patients with pulmonary infection after acute cerebral infarction were randomly allocated to treatment and control groups, 35 cases each. The control group received routine medication and the treatment group, acupuncture in addition. Pre-treatment and post-treatment National Institutes of Health Stroke Scale (NIHSS) scores and clinical pulmonary infection scores (CPIS) were compared between the two groups. The correlation between the NIHSS score and the CPIS score was observed.Results There were statistically significant pre-/post-treatment differences in the NIHSS score and the CPIS score in the two groups (P<0.05,P<0.01). There were statistically significant post-treatment differences in the NIHSS score and the CPIS score between the treatment and control groups (P<0.05). The correlation between the NIHSS score and the CPIS score was low in the treatment group after treatment (r=0.417,P<0.05).Conclusions Acupuncture plus medication is an effective way to treat pulmonary infection after acute cerebral infarction. It can improve the NIHSS score and the CPIS score in the patients.

Objective To construct the recombinant human metapneumovirus(hMPV) (defined as rhMPV NL/1/00 GFP) in vitro by reverse genetics technique. Methods BSR-T7 cells were transfected using LipofectAMINE 2000 with the full-length cDNA plasmid, and four major protein expressing plasmids, pCITE-N, pCITE-P, pCITE-M2.1 and pCITE-L. After 3 d, cells were subjected to one -80℃ freeze-thaw cycle to prepare lysates. The supernatant of lysate was used to inoculate Vero-E6 cells. After 1-4 d, cells were found for the obvious development of cytopathic effects under light microscope and green fluoroscopic signals under fluorescence microscope, and were observed up to 10 d. The supernatant were collected to de-tect virus titer. Viral RNA was extracted from the supernatant and reverse transcriptase polymerase chain re-action (RT-PCR) was used to amplify N, F and G genes of rescued virus. Results Cytopathic effects and green fluoroscopic signals was readily and obviously observed after 1-4 d post-inoculation in Vero-E6 cells, then cytopathic effects got worse and green fluoroscopic signals became stronger gradually up to 10 d. The ti-ters of the 1st, 5th, 10th,15th and 20th generation virus ranged from 105.0 to 106.5 TCID50/ml. Amplicons with size of 910 bp (N), 450 bp (F) and 980 bp (G) by RT-PCR were accordant with expectant. Nucleotide sequence analysis of above cDNA fragments showed 100% similarity with reported sequence of hMPV NI/1/00 strain. The recombinant virus was genetically constant and GFP-labeled after 20 passages in Vero-E6 cells. Conclusion Recombined hMPV was successfully rescued by reverse genetics technique. This study lays ground for exploring pathogenesis of hMPV infection and development of hMPV attenuated vac-cines.

Aim To study the anti-inflammatory activity of the Channa argus bile.Methods The bile was isolated and purified by extraction and silica gel column chromatography.Then the compounds were identified by hydrogen and carbon spectra.The spleen lymphocytes proliferation assay and Lipopolysaccharide(LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide(NO) experiment were used to evaluate the anti-inflammatory activity.Results Compound(C1) of sodium taurocholate and compound(C2) of sodium taurochenodeoxycholate were isolated by activity tracing.The cell relative viabilities of the two compounds on Concanavalin A(Con A) induced spleen lymphocytes proliferation assay were 65.9%±11.7% and 60.5%±9.4%, which were significantly different from the result of model group (P<0.01), respectively.The NO production of LPS-induced RAW264.7 release of NO was (16.4±1.9) μmol·L-1 and (15.5±1.7) μmol·L-1, which were significantly different from the result of model group(P<0.01).Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.

Objective To introduce the mutated gene coding cysteine into the gene of dsFv VL of human antibody to N terminal fragment of lipopolysaccharide binding protein(LBP)and to express,purify the mutated dsFv VL in bacterium.Methods We reconstructed and sequenced the mutated gene of VL of human mAb Fab to LBP by Mega-primer PCR based on point mutagenesis method.Some codes of FWR1 of VL had been replaced by TGT in order to code cysteine.The DNA sequence of reconstructed VL was inserted into vector pET-28a(+),then VL was expressed by E.coli.BL21 star(DE3)and was purified by chromatography.Finally the activity of VL to bind NH-LBP was determined by ELISA.Results The results showed that the cysteine was introduced into the position 21 amino acid of VL to replace the threonine.The gene of VL was about 650 bp and relative molecular weight of VL was 28?103.VL could bind NH-LBP directly.Conclusion These have laid a foundation for producing the dsFv against NH-LBP.