AIM To study the effect of 17?-estradiol(E 2) on apoptosis of macrophages at certain concentration.METHODS Fluorescent microscopy and scanning eletron microscopy(ESEM) were used to detect apoptosis changes of macrophages induced by E 2 at relatively high concentration and intracellular Ca 2+ and matochontrial transmembrane potential were examined by laser scanning confocal microscopy(LSCM). RESULTS Apoptosis of macrophages was induced by high concentration of E 2 (≥1 ?mol?L -1). Intracellular Ca 2+ elevated and matochontrial transmembrane potential decreased after E 2 treatment. CONCLUSION Apoptosis of macrophages is induced by certain concentration of E 2 (≥1 ?mol?L -1) and this may be related to elevated intracellular Ca 2+ and decreased matochontrial transmembrane potential.

ObjectiveTo investigate the effects of dexmedetomidine on postoperative cognitive function and monocytes Toll-like receptor 2 (TLR2)and TLR 4 expression in elderly patients.MethodsForty-five ASA Ⅰ or Ⅱ elderly patients aged ≥65 yr weighing 53-72 kg were randomly divided into 3 groups: control group (group Ⅰ ) and different doses of dexmedetomidine groups(groups Ⅱ and Ⅲ ).Dexmedetomidine 1.0 μg/kg was injected iv over 15 min after anesthesia induction,and then was infused at a rate of 0.5 μg·kg-1 ·h-1 (group Ⅱ ) or 1.0 μg· kg-1 ·h-1 (group Ⅲ ) untile the end of operation.Group Ⅰ received equal volume of normal saline.Blood samples were taken before anesthesia induction,at 1.5 h after the beginning of operation,at the end of operation and at 24 h after operation(T,-T5 ) for determination of monocytes TLR2 and TLR4 expression by flow cytometrybased method.Postoperative cognitive function was evaluated at 1 d before and 7 d after operation with Mini-mental state examination and Wechsler memory scale and Wechsler adult intelligence scale,and the postoperative cognitive dysfunction was recorded.ResultsThe incidence of postoperative cognitive dysfunction and monocytes TLR2 and TLR4 expression were significantly lower in groups Ⅱ and Ⅲ than in group Ⅰ,and in group Ⅲ than in group Ⅱ (P ＜ 0.05).ConclusionDexmedetomidine can prevent postoperative cognitive dysfunction in elderly patients,and the mechanism may be related to down-regulation of monocytes TLR2 and TLR4 expression.

Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.

Objective To detect the impact of Diltiazem on the major adverse cardiac events (MACE) in six months after percutaneous coronary intervention (PCI).Methods A total of 192 patients after PCI with coronary atherosclerotic heart disease were enrolled in this study.The patients were randomly divided into Diltiazem therapy group (101 patients) and non-Diltiazem therapy group (91 patients).The high-sensitivity C-reactive protein (hs-CRP) was assessed before and 24 h after PCI,and the incidence of Major adverse cardiovascular events(MACEs) were assessed at the sixth month after PCI.Results Compared with before PCI,hs-CRP level increased significantly in both group after PCI (P＜0.01),but hs-CRP level was lower in Diltiazem therapy group than in non-Diltiazem therapy group (P＜0.05).Compared with non-Diltiazem therapy group,there was lower incidence of MACEs during six months follow-up in Diltiazem therapy group.Conclusions Diltiazem can decrease the incidence of MACEs during six months after PCI.

Objective:To investigate the clinical efficacy of Zhang's acupoint pressure therapy plus electroacupuncture (EA) in treating post-traumatic knee osteoarthritis.Methods:A total of 98 eligible patients with post-traumatic knee osteoarthritis were divided into group A and B by the random number table, 49 cases in each group. Group A was intervened by Zhang's acupoint pressure therapy plus EA; group B was given medicinal fumigation. The clinical efficacies of the two groups were compared.Results:The markedly effective rate of group A was significantly higher than that of group B.Conclusion:Zhang's acupoint pressure therapy plus EA can produce a satisfactory clinical efficacy in treating post-traumatic knee osteoarthritis, and is worth promotion.

With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.

Objective To test the effect of recombinant human growth hormone (rhGH) on the radiotherapy sensitivity of colorectal cancer cell line, and to explore its relationship with apoptosis. Methods Flow cytometry and immunofluorescence were used to detect growth hormone receptor(GHR) expression on 9 human colorectal cancer cell lines. The colony forming assay was performed to measure the post-radiotherapy colorectal cancer cell proliferation as an indicator of radiotherapy sensitivity. Flow cytometry (Annexin V-FITC staining) was used to detect the radiotherapy induced apoptosis; Westem blot was performed to detect the phosphorylated Akt level within the same cell lines. Results HCT-8 GHR positive expression cell and LoVo GHR negative expression cells were selected for further studies. The colony formation rate was significantly enhanced in HCT-8 cells pre-incubated with thGH as compared to the radiotherapy group ceUs and in a dose dependent manne(0-100 mg/L); under high doses (8 Gy), this effect was more dramatic (52.1±2.9 vs 21.0±2.7, P<0.001). thGH pre-incubation also reduced radiotherapy induced HCT-8 cell apoptosis (P<0.05). When GHR was blocked with neutralizing antibodies, this protective effect was eliminated. By contrast, thGH pre-incubation did not change the colony formation rate in GHR negative expression LoVo cells. GH rapidly induced Akt phosphorylation in HCT-8 cells by GH/GHR signaling, and this effect was blocked by PI-3 kinase inhibitor. Conclusions The protective effect of GH on colorectal cancer cells may occur after radiotherapy exposured by GHR, which is related to the reduction of apotosis.

Objective To isolate and identify 3 flavonoids (taxifolin, orobol and quercetin) from Cudrania tricuspidata, and develop a method for determining 3 flavonoid constituents in Cudrania tricuspidata. Methods Three flavonoids was isolated from ethanol extract of Cudrania tricuspidata by chromatography, and its structure was identified by nuclear magnetic resonance. The analysis was conducted on an Aglient C18 column (4.6 mm ×250 mm, 5 μm) eluted with 1％ acetic acid and methanol as mobile phases in gradient mode. The flow rate was 1 ml/min and the detection wavelength was set at 310 nm. The column temperature was 25 ℃. Results Taxifolin, orobol and quercetin were isolated from ethanol extract of Cudrania tricuspidata by chromatography. The content of taxifolin, orobol and quercetin were 0.850 mg/g, 0.518 mg/g, 0.103 mg/g. Conclusion The method can be used for the quality control of Cudrania tricuspidata as a reference.

Objective To analyze the alkaloids in the different parts of Aconitum wulingense by HPLC-ESI-Trap-MS. Methods The Agilent XDB-C18(250 mm×4.6 mm, 5 μm) column with gradient elution of 0.1% solvent (A)-acetonitrile(B), at a flow rate of 1 ml/min was used. The column temperature was set at 30 ℃. The MS analysis was based on positive ions mode. Results In the roots, a total of 61 diterpenoid alkaloids were discovered, among which 46 were identified. In the stems, 38 alkaloids have been found, among which 33 alkaloids were identified and 27 were the same with the roots. In the leaves, 18 alkaloids have been detected and 8 were the same with the roots. Conclusions The method is accurate, reliable and efficient, and is suitable for rapid identification of ingredients in Aconitum wulingense, which provides a reference for the development and utilization of Aconitum wulingense and clarify its efficacy and material basis.