Objective To evaluate the effect of subfascial endoscopic perforating veins ablation in treating chronic venous insufficiency of lower extremities. Methods A retrospective study was carried out on 18 patients( 20 limbs) with chronic venous insufficiency treated by subfascial endoscopic perforating veins ablation. Results 69 perforating veins were detected in the medial calf of 20 limbs,including 62 incompetent perforating veins and 7 competent perorating veins. 65 perforating veins were ligated but 4 were not found.Apart from the clinical score of pigmentation, there were sigificant decreases in all the mean scores postoperatively (P

Objective To investigate the changes of SCD 14, TNF-?, E-SLT and IL-10 level in the process of infection.Methods Serum E-SLT, IL-10, SCD 14 and TNF-? level was measured in 37 patients of abdominal trauma, and in model rabbits with endotoxemia.Results Serum level of SCD 14,TNF-?,E-SLT and IL-10 on the 1st to 3rd day post-op increased significantly in patients suffering from post-op infection 〔(1.61?0.47)??g/ml, (28.63?8.29)?pg/ml,(153.6?48.9)?ng/ml and (38.21?10.87)?pg/ml, compared with control, all P

Objective To explore the effects of overexpression of hypoxia inducible factor 1? (HIF 1?) mRNA on vascular endothelial growth factor (VEGF) and novoendotheliasis in venous autografts Methods Wistar rats were randomly divided into two groups with 28 rats in each group A rat experimental model of autogenous vein graft was established by transplanting the right external jugular vein into between the interrupted right common carotid artery The transplanted vein in the experimental group was first immersed into a solution containing recombinant adeno associated virus (rAAV) HIF 1? for 45 minutes Vein grafts and blood simples were taken at 7 or 14 days after transplantation RT PCR, ELISA, immunohistochemistry were used to detected HIF 1? mRNA and VEGF expression Results HIF 1? mRNA and VEGF protein remarkably increased in experimental group, and serum level of E selectin significantly decraesed at day 14 The novoendotheliasis and myo endothelium junction in vein grafts were remodeled at day 14 in the experimental group Conclusion Re establishment of the structure and function of the autograft vein graft endothelium was accelerated by overexpressed HIF 1? mRNA

Objective To investigate the mechanism of fluoroquinolone resistance in clinical isolates of Enterococcus faecium. Methods The MICs of six fluoroquinolones(norfloxacin,ciprofloxacin,ofloxacin,levofloxacin,gatifloxacin and moxifloxacin) against 35 clinical isolates of E.faecium from eight hospitals in Tianjin were determined by agar dilution method in the absence or presence of multidrug resistance efflux pump inhibitor reserpine.The quinolone-resistance determining region(QRDR)of parC and gyrA were amplified and sequenced.Results No less than twofold decrease in MIC values of the six fluoroquinolones in the presence of reserpine was observed in 35,29,1,0,6 and 2 of the 35 strains of E.faecium respectively.One fluoro- quinolone-susceptible isolate and five fluoroquinolone-resistant isolates were selected randomly to analyze the QRDR of parC and gyrA.All five fluoroquinolone-resistant isolates had single amino acid alteration in both GyrA and ParC.Ser-80 in ParC was substituted by lie(4 isolates)or Arg(1 isolates).Glu-87 in GyrA was replaced by Lys(2 isolates)or Gly(2 isolates). The other one had an Ser-83-to-Ile substitution.The one fluoroquinolone-suseeptible isolate had no alteration in the QRDR of either ParC or GyrA.Conclusions Both target alteration and active efflux are responsible for the resistance to fluoroquinolone in clinical isolates of E.faecium.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; pathology ; Biopsy ; Biopsy, Fine-Needle ; Child ; Cytodiagnosis ; methods ; Diagnosis, Differential ; Female ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; pathology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Lymphoma, T-Cell ; pathology ; Male ; Middle Aged ; Pleural Effusion ; pathology ; Young Adult

OBJECTIVETo investigate the expressions of stromal cell-derived factor-1 (SDF-1) and CX chemokine receptor-4 (CXCR-4) in patients with hepatocellular carcinoma (HC) and liver cirrhosis.

METHODSPeripheral blood and/or ascites fluid were collected from 39 hepatocellular carcinoma patients, 16 patients with liver cirrhosis, 12 with hepatitis and 12 healthy donors. The SDF-1 expression was assayed by ELISA and CXCR-4 was measured by immunohistochemical methods.

RESULTSThe level of SDF-1 expression in the carcinoma patients was higher than that of the liver cirrhosis, hepatitis patients and healthy donors, but there was no significant difference between those of the healthy donors and hepatitis patients or liver cirrhosis patients. The levels of CXCR-4 expression were closely related to the tumor differentiation.

CONCLUSIONThe expression of SDF-1 in the peripheral blood and the CXCR4 expression in the HCC tissues of the HC patients may be regarded as markers of HC and they may have a positive relationship with the differentiation and metastasis of HC.

Adult ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Chemokine CXCL12 ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; pathology ; Neoplasm Staging ; Receptors, CXCR4 ; metabolism

OBJECTIVETo clone the glial cell line-derived neurotrophic factor (GDNF) from the mouse testis, construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the gdnf-transfected Sertoli cells as the feeder layer to cultivate spermatogonial stem cells (SSCs).

METHODSTotal RNA was extracted from the testes of normal mature mice and gdnf was cloned and amplified using RT-PCR, inserted into the eukaryotic expression vector and transfected into sertoli cells (TM4 cell line). Immunofluorescence with anti-GDNF antibodies was performed at 40 h following the transfection.

RESULTSgdnf cDNA was cloned successfully, and GDNF expressed after transfected into Sertoli cells.

CONCLUSIONThis study provides a basis for culturing SSCs with gdnf-transfected Sertoli cells as the feeder layer.

Animals ; Cloning, Molecular ; Gene Expression ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; Male ; Mice ; Mice, Inbred Strains ; RNA ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; metabolism ; Testis ; cytology ; metabolism ; Transfection

OBJECTIVETo construct an eukaryotic expression plasmid containing gp120 gene of HIV-1 subtype B and obtain gp120 gene expression in HepG2 cells.

METHODSAccording to the published gp120 gene sequence in Genbank, a pair of primers was designed and synthesized. The PCR amplification product of gp120 gene was cloned into pMD-18T vector using TA cloning followed by BamHI and XhoI digestion and sequence analysis. The target gene was then subcloned into a highly efficient eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid was sequenced and identified by restrictive endonuclease digestion, and transfected into HepG2 cells via liposome. The expression of gp120 gene was analyzed by RT-PCR and Western blotting, respectively.

RESULTSRestriction endonuclease digestion and sequence analysis verified successful construction of the recombinant vector pcDNA3.1(+)/gp120. The target fragment gp120 was identical with U26942 in Genbank, and the expression of gp120 gene was detected in the lysate of the transfected HepG2 cells by RT-PCR and Western blotting.

CONCLUSIONThe eukaryotic expression plasmid for gp120 has been constructed successfully, which is capable of stable expression in HepG2 cells.

AIDS Vaccines ; biosynthesis ; genetics ; Base Sequence ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Gene Expression ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; HIV-1 ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Transfection ; Vaccines, DNA ; biosynthesis ; genetics

OBJECTIVETo detect HBV DNA and its genotypes.

METHODSThe 6 isoforms of HBV DNA was detected out using of different probes by Polymerase Chain Reaction and Nucleic Acid hybridization.

RESULTSOf 150 HBV DNA positive patients who lived in Shenzhen, 50 samples (33%) are type B, 36 samples (24%) are type C, 13 samples (9%) are type D, 3 samples is type F, 1 sample is type A, 48 samples (31%) are mixed type. The ALT value was significantly higher in genotype B than in genotype C. HBe positivity were higher in genotype B than genotype C. HBeAg positivity were higher in genotype C than in genotype B. There are not obvious relations between genotype and age or sex.

CONCLUSIONIn the detected samples, the major genotype of HBV DNA is type B, several are type C, D. The type E haven't been found. There are some relations between all kinds of genotypes and the severity of hepatitis B.

Adult ; DNA, Viral ; analysis ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction

AIM AND METHODSBased on the reaction that 2',7'-dichlorodihydrofluorescein (DCFH) can be oxidized by reactive oxygen species (ROS) to yield the highly fluorescent 2',7'-dichlorofluorescin (DCF), ROS production in mitochondria can be observed dynamically as well as quantified directly by spectrofluorometer.

RESULTS AND CONCLUSIONDCF fluorescence showed linear increase in State 4 mitochondria, which suggest ROS produced at constant rate. The slopes of the linear increase in fluorescence with time were computed performing a linear regression that took into account all relevant data points in selected time windows. The slopes were proportional to ROS production in mitochondria. Addition of sodium azide and malonic acid increased and decreased the rate of ROS production respectively during measurement. DCF fluorescence varied linearly with increasing concentration of mitochondria, which showed quantitative relations in definite range. Repeated measures showed low coefficients of variation. This method is reliable and efficient for determining ROS in mitochondria.

Animals ; Fluoresceins ; Fluoroimmunoassay ; methods ; Male ; Mice ; Mice, Inbred Strains ; Mitochondria ; metabolism ; Reactive Oxygen Species ; analysis