Objective To evaluate the efficacy of double flap reconstruction after craniectomy through transpetrosal-presigmoid approach in the treatment of petroclival tumor.Methods A total of patients with petroclival tumor were enrolled in this study.Craniectomy was performed on them through the transpetrosal-presigmoid approach,and then double flap reconstruction was carried out.After the temporo-occipotal free osseous flap(retrosigmoid flap)was obtained,the superficial flap of the mastoid process(presigmoid flap)was freed by grinding and drilling.During the operation,partial petrosectomy and mastoidectomy were avoided to protect the semicircular canals and cochlea.Results Among the 14 cases,radical resection of the tumor was performed on 8 patients,subtotal resection on 3,and partial resection on 3.Two patients developed CSF leak through the ear.No subcutaneous hydrops or intracranial infection was found in the patients.The patients were followed up at 3 and 6 months after the operation,during which no complications were detected.Conclusions The rates of CSF leak,subcutaneous hydrops,and intracranial infection are low after applying double flap reconstruction to craniectomy through the transpetrosal-presigmoid approach.The procedure are mini-invasive and safety.

Objective To investigate effect of pidotimod on TH1/TH2 balance and immune factor in patients with severe trauma.Methods 90 cases with severe chest trauma patients were randomly divided into two groups.45 cases in control group were treated by conventional anti-inflammatory treatment, 45 cases in experimental group on base of control group with pidotimod 800mg /times, 1 times daily.After treatment, the serum levels of IL-4, IFN-γ, IL-2, IL-10, IL-17,IgM, IgA and IgG were compared.Results Compared with before treatment, indexes were improved(P<0.05). Compared with the control group, IL-4 level was lower, IFN-γlevel was higher, IFN-γ/IL-4 was higher(P<0.05).IL-2, IL-10, IL-17 level was higher(P<0.05).IgA, IgM, IgG level was higher(P<0.05).Conclusion Pidotimod can balance the TH1/TH2 in the patients with severe trauma, and adjust the serum levels of IgM, IgA and IgG,can be used in clinical application.

Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.

Recombinant expression vector pcDNA3-MCPCD59DP containing human membrane complement regulatory proteins(hCRPs) MCP and CD59 cDNA was constructed successfully by using two independent promoters.After transfected into NIH3T3 cells with calcium phosphate-DNA precipitate method,NIH3T3 pcDNA3-MCPCD59-DP transfectants were obtained by G418 selection.Extraneous genes integration was identified by PCR.The co-expression of human CD59 and MCP at both mRNA and protein levels was confirmed by using RT-PCR and Western blot analysis.Human MCP and CD59 cDNA were integrated in NIH3T3 pcDNA3-MCPCD59-DP genomic DNA after continuous 30 times passages,indicating that NIH3T3 pcDNA3-MCPCD59-DP were stable cell lines.Human complement-mediated cytolysis assays showed that NIH3T3 cells transfected stably with pcDNA3-CD59,pcDNA3-MCP,and pcDNA3-MCPCD59-DP were protected from C-mediated damage and co-expressed human MCP and CD59 provided more excellent protection against C-mediated attack as compared with either CD59 or MCP expressed alone.The dicistronic vector represents an effective and efficacy strategy to overcome C-mediated damage and has potential therapeutic value for effectively controlling complement activation and finally for preventing hyperacute rejection in clinical gene therapy.

Objective To investigate the role of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) in inducing insulin resistance in differentiated human adipocytes.Methods Human preadipocytes obtained via liposuction were induced to differentiate into mature adipocytes.iE-DAP,a specific ligand for NOD1,was administered to human adipocytes in culture.NF-κB transcriptional activity and proinflammatory cytokines production were determined by luciferase assay and enzyme-linked immunosorbent assay.Glucose uptake in adipocytes was measured by 2-deoxy-D-[3 H] glucose uptake (P＜0.05).The expression of phosphatidylinositol-3-kinase p85 (PI-3K p85),insulin receptor substrate-1 (IRS-1) and Akt phosphorylations were detected by Western blotting.Results NF-κB transcriptional activity and cytokines such as interleukin (IL)-6,IL-8,and monocyte chemotactic protein 1 secretion were markedly increased after stimulation with iE-DAP (P＜0.05 or P＜0.01).Insulin-induced glucose uptake was decreased with the activation of NOD1 in a dose-and time-dependent fashion.NOD1 activation weakened insulin signal transduction as being revealed by increasing IRS-1 Ser307phosphorylation,reducing protein expression of PI-3K p85,and attenuating insulin-induced phosphorylation of Akt on Ser473 and Thr308in human adipocytes.Conclusion These results indicate that NOD1 activation induces inflammatory response and insulin resistance via IRS-1/PI3-K/Akt signaling pathway in human adipocytes.

Objective To establish a method for determination the contents of six phendic acids in Mailuoning injection by HPLC at same time. Methods Separation was performed on a Kromasil-C18 (4.6 mm? 250 mm, 5 ?m) column with mobile phase consisting of 0.1% phosphoric acid-water and Acetonitrile with gradient elute at the flow rate of 1 mL/min. The UV detection wavelength was set at 327 nm and the column temperature was set at 25 ℃. The sample size was 20 ?L. Results The standard cures of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 3,5-Dicaffeoylquinic acid, 3,4- Dicaffeoylquinic acid were linear within the contentration range of 0.010 90～0.544 8 ?g (r=0.999 9), 0.024 06～1.203 2 ?g (r =1), 0.011 06～0.552 8 ?g (r =1), 0.015 94～0.796 8 ?g (r =1), 0.009 104～0.455 2 ?g (r =1), 0.010 37～0.518 4 ?g (r=1) respectively. And the recovery rates of them were 101.06%, 100.22%, 101.54%, 99.42%, 100.21%, 101.65% respectively. Conclusion The method appeared to be simple, reliable, accurate and can be used to determine the contents of six phendic acids in Mailuoning injection.

Aim To study the influence of Liensinine(LIE) on the oxidative damage of erythrocyte ghost. Method The erythrocyte ghost oxidative injury caused by super oxide anion (O?_2) from the auto-oxidation of pyrogallic acid was used as model. The effect of LIE on oxidated erythrocyte ghost was observed by the measurement of the malonidial aldehyde (MDA)-the degradation product of lipid peroxidation, blocking degree, speed of auto-oxidation and membrane fluidity. Result O?_2 caused lipid peroxidation of erythrocyte ghost, increased the content of MDA and the speed of auto-oxidation, decreased membrane fluidity, and alleviated blocking degree. But LIE(80, 100, 120 mg?L -1 )could decrease the content of MDA and speed of auto-oxidation markedly, improve the membrane fluidity and blocking degree significantly, and inhibit the lipid peroxidation. Conclusion LIE has protective action on the oxidative damage of erythrocyte ghost initiated by O?_2

Objective To express and purify the killer immunoglobulin-like receptor KIR3DL1 extracelluar domain.Methods pUC57-KIR3DL1 was used as template,KIR3DL1 extracelluar domain was amplified by polymerase chain reaction(PCR)and cloned into pGEM-T vector with A-T cloning technique.After DNA sequence analysis,the target fragment was inserted into the prokaryotic expression vector pET28a-DsbA to construct the recombinant vector pET28a-DsbA /KIR3DL1.The recombinant plasmid was transformed into E.coli BL21(DE3),and induced with IPTG.Bacterial pellets were resuspended in 8M urea and centrifuged to remove the insoluble material.The crude extract was purified by passing over a Ni-NTA-agarose column.After the inclusion body flowing through the Ni-NTA-agarose affinity chromatography was refolded successfully,it was purified by Superdex75 gel filtration and the purification effects of the fusion protein were identified by SDS-PAGE and western blot.Result Some fusion proteins were expressed in the supernatant,and the others were expressed in the form of inclusion bodies.The purity of fusion protein was over 95% after purification under denaturing condition.Conclusion The highly efficient expression of KIR3DL1 extracelluar domain laid the foundation for the further studies on exploration of the mechanism of immunization recognition between KIR3DL1 and its ligand.

Objective To characterize the relationship between diabetic peripheral neuropathy(DPN) and serum anti gangliosides antibody(GS). Methods Elderly patients were divided into three groups: patients with type 2 diabetes mellitus(DM) showing no DPN,DM group, 25; DPN group, 32; healthy control group, 30. The nerve conduction velocity was measured using Nicolet Viking type IV electromyography(USA). The anti GSIgM Ab and anti GSIgG Ab were examined by solid enzyme immunoassay, and interleukin 1?(IL 1?), IL 2,tumor necrosis factor ?(TNF ?), HbA 1C and NO were also measured. Results The positive rate of anti GSIgM Ab and anti GSIgG Ab in DPN group being 65.6% and 34.4% respectively, was obviously higher than those in N group and DM group(P

OBJECTIVE:To observe therapeutic efficacy and safety of low-molecular-weight-heparin-sodium combined with dy-drogesterone in the treatment of threatened abortion. METHODS:Medical information of 72 patients with threatened abortion were analyzed retrospectively and divided into control group(36 cases)and observation group(36 cases). Control group was given Dy-drogesterone tablet with initial dose of 40 mg,one day later every 12 h 10 mg/time. Observation group was additionally given ab-dominal subcutaneous injection of Low-molecular-weight-heparin-sodium injection 5000 U,once a day,on the basis of control group. Both groups were treated for a week. Clinical efficacies of 2 groups were observed,and lumbar acid,vaginal bleeding,ab-dominal pain time,total treatment time,neonatal birth weight were also observed. The levels of hs-CRP,fibrinogen,D-dimer and platelet,the occurrence of ADR were observed before and after treatment. RESULTS:The total response rate of observation group was significantly higher than that of control group(91.7%vs. 80.6%),the backache,vaginal bleeding,abdominal pain time and to-tal treatment time of observation group were significantly shorter than those of control group,the neonatal birth weight and gesta-tional age of observation group were significantly more than those of control group,with statistical significance(P<0.05). Before treatment,there was no statistical significance in the levels of hs-CRP,fibrinogen,D-dimer and platelet between 2 group(P>0.05);after treatment,the levels of hs-CRP,fibrinogen,D-dimer and platelet in 2 groups were significantly lower than before,and the observation group was significantly lower than the control group,with statistical significance (P<0.05). There was no statistical significance in the incidence of ADR between 2 groups (P>0.05). CONCLUSIONS:Based on routine treatment,low-molecu-lar-weight-heparin-sodium combined with dydrogesterone shows good therapeutic efficacy and safety for threatened abortion.