Aim To study gyrA and parC mutations of clinical Pseudomonas aeruginosa strains. Methods MIC values of 55 clinical P.aeruginosa isolates were determined by agar dilution test and 1 sensitive strain and 8 resistant strains were selected with standard sensitive strain ATCC27853 as control, the quinolone determining region (QRDR) of the gyrA and parC genes were amplified by PCR, the lengths of PCR products were 351 bp and 397 bp. The gyrA PCR products(351 bp) were digested with enzyme sacⅡ. The gyrA and parC gene were sequenced. Results In this study, gyrA genes of all resistant strains had an ACC to ATC mutation in codon 83, leading to the amino acid substitution of an isoleucine for a threonine, and three high level resistant strains also showed a GAC to GGC mutation in codon 87, leading to the substitution of a glycine for an aspartic acid. In addition, four resistant strains also had an TCG to TTG mutation in codon 87 of parC gene, leading to the amino acid substitution of a serine for a leucine. The strains with both gyrA and parC mutations were two to sixteen times more resistant than the strains which had only gyrA mutations. At the same time, a silent mutation (CAC to CAT) in codon 132 of gyrA gene and a silent mutation(GCT to GCG) in codon 115 of parC gene occured, which did not lead to amino acid change. Conclusion The mutations of 83 and 87 codons of gyrA and the mutatations of 87 codon of parC gene were related to fluroquinolone resistance, and the mutations of the 83 codon of gyrA gene were more important.

Objective 22q11 microdeletion is featured with hemotological dysfunction,among others:hypocalcemia,thrombocytopenia and megathrombocyte.This study was trying to work out the platelet characteristics of 22q1 1 microdeletion patients and its potential clinical application.Methods A total of 80 cases who had undergone open-heart surgery were selected and confirmed with fluorescence in situ hybridization (FISH) to have 22q1 1 microdeletion among 40 cases while the others were negative.The relevant data in full blood routine of all the 80 cases in hospital information system (HIS) then were collected and analyzed with suitable statistical methods.Results The mean platelet volume in microdeletion group was statistically higher than that in the control group [(11.20 ± 1.94)fL vs (8.95 ± 1.58) fL,P ＜0.01].The area under the receiver operating characteristic (ROC) curve was 0.82,meaning significant predictive values.The corresponding sensitivity and specificity for mean platelet volume (MPV) =10 fL were 70.0 ％ and 80.0 ％,respectively.Conclusions MPV in congenital heart defect patients with 22q11 is significantly higher than those without 22q11.It is an effective method for preliminary screening 22q11.Being obtained from full blood routine data,it is economic and quick.MPV =10 fL can be used as a cutoff for guidance for irradiated blood transfusion postoperatively.

In this paper, a qualitative and quantitative analysis method was established for the scopoletin in herba Artemisiae hedinii hy HPLC. The chromatographic column was Agilent TC-C18 (4.6 mm×250 mm, 5μm);the tem-perature of column was 30℃, eluted with a mobile phase consisted of methanol and 0.3%phosphoric acid solution and gradient elution at a flow rate of 1.0 mL·min-1. The detection wavelength was 344 nm. The results showed that the mass concentration and peak area of scopoletin had a good linear relationship within the range of measurement (r=0.999 9). The reproducibility was good;and the RSD was 1.51%. The average sample recovery rate was 101.42%. It was concluded that the established method was stable, reliable and simple. It provided theoretical evidences for the quality evaluation and quality control of herba A. hedinii.

BACKGROUND: Effect of hypoxia on osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) has been differently reported. Those differences might cause by varying volume fraction of oxygen and varying source of BMMSCs. OBJECTIVE: To compare the biological differences between placenta and BMMSCs under hypoxia.METHODS: Human placenta amniotic mesenchymal stem cells (hAMSCs) and rabbit BMMSCs were isolated by two step proteinases and whole bone marrow adhesion, respectively. hAMSCs and BMMSCs at the same passage were seeded in 12-well plates at an initial cell density of 2 × 10~4 cells per well with α-MEM containing 10% FBS. Then, the cells were cultured under 5% O_2 or 20% O_2 for 12 days. hAMSCs and BMMSCs at the same passage were seeded in 12-well plates at an initial cell density of 1 × 10~5 cells per well with osteogenic medium. Then, the cells were cultured under 5% O_2 or 20% O_2 for 14 days. Cell growth curve, the specific glucose consumption rates and specific lactate production rates, and osteogenic differentiation were detected. RESULTS AND CONCLUSION: Compared to normal oxygen, hypoxia promoted the proliferation and osteogenic differentiation of MSCs. When compared to BMMSCs, statistically significant enhancement of the growth of hAMSCs by hypoxia was observed. hAMSCs cultured under hypoxia exhibited lower glucose consumption and lactate production in contrast with BMMSCs. Furthermore, comparison between hAMSCs and BMMSCs showed that the alkaline phosphatase expression of BMMSCs was significantly enhanced by hypoxia and was markedly higher compared with hAMSCs. The amount of calcium deposition was also enhanced by hypoxia, but there were no statistically significant differences between hAMSCs and BMMSCs.

Objective To evaluate the value of CT Angiography (CTA)in the diagnosis of cerebral aneurysms in patients with a-cute subarachnoid hemorrhage (SAH )with Meta analysis.Methods A systematic search in PubMed,MEDLINE,EMBASE, CNKI,CBM,WANFANG databases was performed.Criteria for inclusion were established based on validity criteria for diagnostic research published by the Cochrane Methods Group on Screening and Diagnostic.The methodological quality of each study was eval-uated by QUADAS.Meta-Disc Version 1.4 software were used for heterogeneity test and the pooled weighted sensitivity,specifici-ty,positive likelihood ratio,negative likelihood ratio and 95% confidence intervals were calculated,receiver operating characteristic (SROC)and area under the curve(AUC)were obtained with random-effects models.Stata12.0 was used for the funnel plot to ana-lyze publication bias.Results Fourteen studies were included with a total of 1 332 patients and 1 149 ruptured cerebral aneurysms. The pooled weighted sensitivity and specificity of CTA with 95% confidence interval was 0.96(0.95-0.97)and 0.99(0.97-1.00) respectively.The AUC and the Q value of SROC was 0.99 and 0.958 4 respectively.Conclusion CTA has high sensitivity and spe-cificity in the diagnosis of cerebral aneurysms in patients with SAH,can be proposed as an important imaging diagnostic method.

OBJECTIVE: To evaluate the inhibiting effects of the root of Mallotus apelta (Lour.) Muell.-Arg. on duck hepatitis B virus (D-HBV) in vivo. METHODS: Forty nestling ducks with congenitally infection of D-HBV detected by PCR were randomly divided into five groups: untreated group, lamivudine-treated group, and high-, medium- and low-dose root of Mallotus apelta-treated groups. The ducks in the lamivudine-treated group were fed lamivudine with a dose of 50 mg/kg once. Ducks in the three-dose Mallotus apelta-treated groups were treated with different doses of decoction of this herbal medicine for 21 days respectively. The serum content of D-HBV DNA was determined by quantitative real-time PCR technique before and 7 days after the treatment, and on the 7th, 14th and 21st day of the treatment. Liver biopsy was also executed before and after the treatment to observe the histopathological changes. RESULTS: Lamivudine showed a rapid inhibiting effect on D-HBV DNA, but this effect didn't last long, and the serum level of D-HBV DNA increased again after treatment. The serum level of D-HBV DNA dropped markedly in the high- and medium-dose Mallotus apelta-treated groups on the 14th and 21st day. Low-dose Mallotus apelta revealed no obvious inhibiting effect on D-HBV. After treatment, the inhibiting effect in the root of Mallotus apelta-treated group continued as compared with that in the untreated group. The histopathological changes of liver tissues showed that the inflammation in the high-dose root of Mallotus apelta-treated group was weakened as compared with that in the lamivudine-treated group. CONCLUSION: The root of Mallotus apelta has therapeutic effect on D-HBV. It can restrain the duplication of D-HBV in vivo. Although this effect is weaker than that of lamivudine, it continues longer. Thus this herbal medicine is an effective, safe and economical drug for hepatitis B.

Objective To observe the effectiveness and mechanism of Xiaoyao San (Xiaoyao Powder for Soothing Liver and Relieving Depression) in improving depression-like behavior of rats. Methods Male Wistar rats were randomized into normal group, model group, Xiaoyao San (1.9 g·kg-1·d-1) group, and fluoxetine (2 mg·kg-1·d-1) group. The rats were exposed to chronic unpredictable mild stress ( CUMS) to induce rat depression-like behavior. Field test was performed for the observation of effect of Xiaoyao San on rat depression-like behavior, Luminex liquid chip system was applied to detect the serum cytokines, and the amount and size of rat hepatic sinusoidal endothelial window were examined under electron microscope, and hepatic indoleamine 2, 3-dioxygenase ( IDO) and tryptophan 2, 3 -dioxygenaes ( TDO) expression levels were detected by immunohistochemical and Western blot methods. Results Xiaoyao San showed obvious effect on increasing sugar water consumption, the number of crossing the blocks and erection frequency in rats, decreasing serum levels of tumor necrosis factor alpha ( TNF-α) and interleukin 6 ( IL-6) , increasing the amount of hepatic sinusoidal endothelial window, promoting hepatic sinusoidal endothelial vascularization, and reducing TDO and IDO expression ( P<0.05 or P<0.01). Conclusion Xiaoyao San exerts obvious effect on improving rat depression-like behaviors, and the mechanism is probably related with the decrease of inflammatory factors, inhibition of IDO pathway, and improvement of hepatic sinusoidal endothelial function.

ObjectiveTo establish rabbit knee joint cartilage injury models to evaluate effects of the type Ⅱ collagen sponge in repair of the articular cartilage.MethodsThe type Ⅱ collagen sponge was prepared according to previous method and the pore size of the sponges was measured based on the collagen autofluorescence characteristics.The type Ⅱ collagen sponge was transplanted into the injury lesions of the animal model for experimental study.The regeneration of the cartilage defects was observed by using MRI, histologic HE staining, Safranin O, sirius red polarized light staining, areas determination of the newly grown cartilage and immunohistochemistry of type Ⅱ collagen.ResultsAutofluorescent images of confocal microscope layer scanning showed that the pore size was (93.26 + 38.40) μm in diameter, suitable for chondrocyte growth.Comparison between MRI and H&E staining results showed quicker effusion absorption in the treatment groups than that in the control group, while the level of inflammatory response in the treatment group was lower than that in the control group.The sporadic cartilage signals first appeared at the 6th week.The newly formed cartilage with the expression of glycosaminoglycan and type Ⅱ collagen matrix was confirmed by Safranin O staining and immunohistochemical analysis.The sirius red polarized light staining showed that areas of the newly formed cartilage were significantly larger in the treatment group than that in the control group (P ＜ 0.01).Conclusion The type Ⅱ collagen sponge developed from purification can effectively repair the damaged cartilage tissues of the rabbit knee joints, as has been verified either by MRI or histology.

Objective: To analyze whether heat-shock protein 90 (HSP90) be involved in a permanently abnormal activated JAK/STAT signaling in ATL cells in vitro. Methods: The effect of 17-AAG on proliferation of ATL cell lines HUT-102 was assessed using CCK8 at different time points. Cell apoptosis was measured by flow cytometry. The specific proteins HSP90, STAT5, p-STAT5 and JAK3 were detected by Western blotting. Results: Overexpression of HSP90 in HUT-102 cell lines was disclosed (P<0.05) , and constitutive activation of JAK3/STAT5 signaling was observed in HTLV-1-infected T-cell lines but not in normal PBMCs; Treatment of ATL cell lines with 17-AAG led to reduced cell proliferation, but there was no significant change in terms of cell proliferation when the concentration of 17-AAG between 2 000-8 000 nmol/L (P>0.05) . 17-AAG induced cell apoptosis in different time-points and concentrations. 17-AAG don’t affect the expression of JAK3 gene. Conclusion This study indicated that JAK3 as HSP90 client protein was aberrantly activated in HTLV-1-infected T-cell lines, leading to constitutive activation of p-STAT5 in JAK/STAT signal pathway, which demonstrated that HSP90-inhibitors 17-AAG inhibited the growth of HTLV-1-infected T-cell lines by reducing cell proliferation and inducing cell apoptosis.

OBJECTIVETo investigate the role of autophagy on the pathogenesis of spinocerebellar ataxia 3/Machado-Joseph disease (SCA3/MJD).

METHODSHEK293 cells expressing polyglutamine-expanded ataxin-3 were used as cell model for SCA3/MJD. The level of polyglutamine-expanded ataxin-3 was detected after cells were treated with different inhibitors or inducer of autophagy.

RESULTSInhibition of autophagy increased aggregate formation and cell death in HEK293 cells expressing mutated ataxin-3, and vice versa.

CONCLUSIONThe data suggested that autophagy is involved in the degradation of mutant ataxin-3, resulting in a decrease in the proportions of aggregate-containing cells and cell death in HEK293 cells expressing polyglutamine-expanded ataxin-3. It is possible that autophagy may be applied as a potential therapeutic approach for SCA3/MJD.

Ataxin-3 ; Autophagy ; Cell Line ; Humans ; Machado-Joseph Disease ; genetics ; metabolism ; physiopathology ; Mutation ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; metabolism ; Repressor Proteins ; genetics ; metabolism