Objective To explore the apoptosis and proliferation of peripheral blood lymphocytes(PBLs) from primary nephrotic syndrome(PNS) and effects of dexamethasone(Dex) on them.Methods Minimal change NS(MCNS), non-minimal change NS(NMCNS) and healthy children were involved in this study. PBLs were cultured in vitro with Dex or PHA+Dex or without PHA and Dex. Apoptosis of PBLs was measured by propidium iodide(PI) staining; The effects of Dex at different concentrons on PBLs′proliferation were investigated by 3H-TdR incorporation.Results The apoptotic rate of vacuity group in MCNS was lower compared with NMCNS and healthy controls(P

Objective To study the expression of membrane glucocorticoid receptors(mGR), the correlation between mGR and glucocorticoid(GC)'s effects on apoptosis and proliferation of peripheral blood lymphocytes(PBLs) in minimal change nephrotic syn-drome(MCNS)and the influence of GC on mGR. Methods MCNS, nonminimal change NS(NMCNS) and healthy children were involved in this study. Indirect immune fluorescence and flow cytometry were used to examine the percentages of positive mGR lymphocytes; the apoptosis of PBLs was measured by propidium iodide(PI) staining and the proliferation of PBLs was investigated by fa -TdR incorporation. Results 1.mGR expression in MCNS was higher than that in NMCNS and healthy control,but it was reduced after clinical GC therapy(P

In clinical practice, we have found that cognitive impairment frequently occurs in chronic obstructive pulmonary disease (COPD) patients, but little is known about its pathophysiological mechanism. Given that brain-derived neurotrophic factor (BDNF) is affected by many factors such as smoking, infection, hypoperfusion and hypoxia, the present study was to explore the expression of BDNF in COPD rats. The rat COPD model was established by passive smoking and intratracheal instillation of lipopolysaccharide (LPS). Rats with the same age and gender ratios were divided into 4 groups: the control group (n = 6), the smoking group (n = 6), the LPS group (n = 6) and the smoking + LPS group (n = 6, COPD model). Level of BDNF in serum was measured by ELISA. And the expression of BDNF in the hippocampus was assessed using the immunohistochemistry and image analysis. The results showed that BDNF in the hippocampus and serum significantly increased in the smoking, LPS and smoking + LPS groups, compared to that in the control group. However, the expression of BDNF was less in the smoking + LPS group than that in the smoking or LPS group both in the hippocampus and serum. In conclusion, the expression of BDNF in the hippocampus and serum is highly increased in the COPD group. Smoking and intratracheal instillation of LPS induce the increase of BDNF level in the hippocampus and serum.
Animals ; Brain-Derived Neurotrophic Factor ; blood ; metabolism ; Disease Models, Animal ; Female ; Hippocampus ; metabolism ; Lipopolysaccharides ; Male ; Pulmonary Disease, Chronic Obstructive ; chemically induced ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Tobacco Smoke Pollution

Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.

Objective To investigate the Wallerian degeneration of neural fiber tract in medulla and bilateral middle cerebellar peduncle following pontine infarction and to explore its impacts on neurological recovery.Methods Fourteen patients with a recent unilateral pontine infarct underwent the diffusion tensor imaging(DTI)and evaluations with the NIH stroke scale(NIHSS),the Fugl-Meyer motor scale(FM), ataxia rating scale(ARS)and the Barthel index(BI)at the first week(W1),the fourth(W4)and twelfth week(W12)respectively.Mean diffusivity(MD)and fractional anisotropy(FA)were measured at pons, medulla and middle cerebellar peduncle.Fourteenth age and gender matched volunteers underwent a DTI were studied as controls.Results Compared with the matched regions in controls,the FA values of infarct side medulla and bilateral middle cerebellar peduncle in patients significantly decreased at W1,W4 and W12(on the infarct side of medulla:W1:0.43?0.01;W4:0.37?0.02;W12:0.30?0.02;on the infarct side of middle cerebellar peduncle:W1:0.50?0.01;W4:0.43?0.02;W12:0.35?0.04;on the opposite side of middle cerebellar peduncle infarction:W1:0.54?0.02;W4:0.52?0.03;W12:0.47?0.04,t values are 1.92 to 28.56,P0.05 respectively).The absolute value of percent reduction of FA in infarct side medulla and bilateral middle cerebellar peduncle was correlated negatively to the absolute value of percent change of NIHSS and BI score(P

Adult ; Age Factors ; Aged ; Anisotropy ; Brain ; pathology ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Hypertension ; pathology ; Male ; Middle Aged ; Sex Characteristics

Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.

Adult ; Aged ; Cardiomyopathy, Hypertrophic ; diagnostic imaging ; physiopathology ; surgery ; Catheter Ablation ; Coronary Circulation ; Female ; Heart ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Radionuclide Imaging

This study was purposed to establish a retrovirus-mediated murine model with MLL-AF9 leukemia, so as to provide a basis for further investigation of the pathogenesis and therapeutic strategy of MLL associated leukemia. Murine (CD45.2) primary hematopoietic precursor positively selected for expression of the progenitor marker c-Kit by means of MACS were transduced with a retrovirus carrying MLL-AF9 fusion gene. After cultured in vitro, the transduced cells were injected intravenously through the tail vein into the lethally irradiated mice (CD45.1). PCR, flow cytometry and morphological observation were employed to evaluate the murine leukemia model system. The results showed that MLL-AF9 fusion gene was expressed in the infected cells, and the cells had a dramatically enhanced potential to generate myeloid colonies with primitive and immature morphology. Flow cytometric analysis revealed that the immortalized cells highly expressed myeloid lineage surface markers Gr-1 and Mac-1. Moreover, the expression levels of Hoxa9 and Meis1 mRNA were significantly higher in the MLL-AF9 cells than that in control. The mice transplanted with MLL-AF9 cells displayed typical signs of leukemia within 6-12 weeks. Extensive infiltration leukemic cells was observed in the Wright-Giemsa stained peripheral blood smear and bone marrow, and also in the histology of liver and spleen. Flow cytometric analysis of the bone marrow and spleen cells demonstrated that the CD45.2 populations expressed highly myeloid markers Gr-1 and Mac-1. The leukemic mice died within 12 weeks. It is concluded that the retrovirus-mediated murine model with MLL-AF9 leukemia is successfully established, which can be applied in the subsequent researches.
Animals ; Disease Models, Animal ; Leukemia, Biphenotypic, Acute ; Mice ; Mice, Inbred C57BL ; Oncogene Proteins, Fusion ; genetics ; Retroviridae ; genetics ; Transfection

OBJECTIVETo study the distribution and drug resistance of methicillin resistant Staphylococcus strains in various specimens of inpatients in burn wards, and to provide reference for clinical treatment.

METHODSBacteria were isolated from specimens of wound exudate, blood, sputum, and bronchoalveolar lavage fluid etc., which were collected from patients hospitalized in our burn wards from January 2008 to December 2010. The bacteria were routinely cultured and identified. Drug resistance of the Staphylococci to 15 antibiotics commonly used in clinic was identified by K-B disk diffusion method. Data were processed with statistical software WHONET 5.5. The homology of 40 strains of methicillin resistant Staphylococcus aureus (MRSA) was analyzed by pulsed-field gel electrophoresis (PFGE).

RESULTSAltogether 386 strains of Staphylococcus were isolated, including 196 strains of Staphylococcus aureus and 190 strains of coagulase negative Staphylococcus. The mean annual isolation rates of MRSA and methicillin resistant coagulase negative Staphylococcus (MRCoNS) were respectively 73.00% (143/196) and 74.20% (141/190). The resistance rates of MRSA and MRCoNS to β-lactams drugs, such as penicillin, oxacillin, cefazolin, and cefuroxime were 100.00% in every year. No Staphylococcus strains resistant to vancomycin, teicoplanin, or linezolid were found. Three different PFGE patterns A, B, and C were identified among 40 MRSA strains, including 33 strains of type A (30 strains in sub-type A1 and 3 strains in sub-type A2), 6 strains of type B (respectively 3 strains in sub-types B1 and B2), and 1 strain of type C.

CONCLUSIONSThe isolation rates of MRSA and MRCoNS were high in our burn wards from January 2008 to December 2010. All of them showed strong drug resistance property, and they were multidrug resistant. The most prevalent strain was PFGE type A.