Objective　To investigate the mechanism affecting on permeability of vascular endothelial cell by nitric oxide (NO). Methods　Series concentration of sin-1（a donor of NO） were added to ECV 304, a cell line of human umbilical vein endothelium. Cell growth and expression of f-actin, a cytoskeleton protein were observed. Results　Cell growth was inhibited with a dose from 6.25 to 100 μmol/L and was caused to death at the concentration of 50 to 100 μmol/L by sin-1. The expression of f-actin was suppressed obviously after cultured with 100 μmol/L sin-1 for 4 hours. Conclusion　It suggests that anomaly increased NO can increase permeability of blood vessels by suppressing the expression of f-actin, inhibiting cell growth or even resulting in cell death.

Adult ; Air Pollutants, Occupational ; adverse effects ; Chemical Industry ; Female ; Humans ; Insecticides ; adverse effects ; Male ; Maximum Allowable Concentration ; Middle Aged ; Occupational Diseases ; chemically induced ; diagnosis ; Organothiophosphorus Compounds ; adverse effects ; Phosphorus Compounds ; adverse effects ; Sulfides ; adverse effects

Background and purpose:Urothelial carcinoma of the bladder (UCB) is the most common cancer in urinary system. Yes associated protein (YAP) gene is closely associated with urothelial carcinoma of the bladder. The study was aimed to explore the effect of siRNA targeting the YAP gene on cell proliferation and migration of T24 cells. Methods:Small interfering RNA (siRNA) was transfected together with LipofectamineTM2000 in T24 human bladder cancer cells to block the YAP signal pathway. The effect of siRNA on cell proliferation and invasiveness was assessed by cell counting kit-8 (CCK-8) assay, Transwell migration assay and wound healing assay. Quantitative real time-Polymerase chain reaction (qRT-PCR) and Western blot analysis were used to conifrm the successful suppression of YAP gene and protein by siRNA. Results:Expression of YAP gene and protein was successfully suppressed after transfected with siRNA which verified by qRT-PCR and Western blot(RNA:F=93.91, P<0.000 1; Protein: F=4.62, P<0.05). As CCK-8 test showed, the proliferation of T24 bladder cancer cells was successfully restrained by inhibition of YAP gene compared with blank control and negative control(12 h: F=6.00, P=0.037;24 h: F=41.72, P=0.000 3;36 h:F=462.8, P<0.000 1;48 h:F=236.6, P<0.000 1;72 h:F=140.5, P<0.000 1). Transwell and wound healing test were performed after YAP gene was interfered by siRNA. The result demonstrated that migration of T24 bladder cancer cells was signiifcantly inhibited (Transwell: F=43.55, P<0.05;Wound healing: F=43.55, P<0.05). Conclusion:This study suggested that YAP gene was an important enhancer for the proliferation and migration of bladder cancer cells.

It was constructed that a genomic DNA library from Lactobacillus sp. MD-1 yielding D, L-lactic acid. The gene encoding L-lactate dehydrogenase (L-LDH) was cloned from the genomic library of strain MD-1 by complementation in E. coli FMJ144 which was lactate dehydrogenase and pyruvate-formate lyase double defective mutant. The nucleotide sequence of the ldhL gene predicted a protein of 316 amino acid starting with ATG. The putative molecular weight of the L-LDH amino acid sequence was 33.84kD. A putative typical promoter (-35 and -10 boxes) had been observed in the 5' noncoding region. An rho-independent transcriptional terminator has been observed in the 3' noncoding region. Three highly conserved regions (Gly13 approximately Asp50, Asp73 approximately Ileul00 and Asn123 approximately Arg154) with several conserved residues had been identified. Gly13 approximately Asp50 was NADH-binding site domain. Asp73 approximately Ileu100 and Asn123 approximately Arg154 were reported to be the active site domains. The ldhL and the L-LDH of Lactobacillus sp. MD-1 showed the low identity and similarity with other Lactobacilli, and the highest percentage were 61.9% and 68.9% respectively. All the above indicated this gene is a novel ldhL.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; L-Lactate Dehydrogenase ; chemistry ; genetics ; physiology ; Lactobacillus ; genetics ; Molecular Sequence Data

Objective To observe the effects of repeated Botulinum toxin type A (BTX-A) injection on lower limb spasticity after stroke.Methods 180 cases with lower limb spasticity after stroke were divided into the treatment group (n=90) and the control group (n=90). The treatment group was treated with BTX-A injection twice in the spastic muscles at interval of 3~6 months, while both the treatment group and the control group accepted the rehabilitation based on the neurodevelopmental therapy. They were assessed with modified Ashworth Scale (MAS), Fugl-Meyer Lower Limb Assessment (FMAL), Berg Balance Scale (BBS), modified Barthel Index (MBI) before each injection, and 3 d, 7 d, 1 month, 3 months after each injection or the same time for the controls. Results There was significant difference in scores of MAS, FMAL, BBS, MBI for the treatment group among before and 3 d, 7 d, 1 month after each injection (P<0.05), but not significant between 2 injections (P<0.05). There was significant difference in scores of all the assessment between the treatment and control group at the same time (P<0.01). Conclusion Repeated intramuscular injection of BTX-A can reduce the spasticity of lower limb after stroke.

Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.

Objective To analyze the MRI features of viral encephalitis in children.Methods MRI data of 33 children with viral encephalitis diagnosed and treated by clinic were analyzed retrospectively for lesion location,shape,characteristics in multi-sequence and dynamic follow-up.Results The positive rate of MRI detection was 93.93%,including 20 cases involved in the limbic system,3 cases involved in the non-limbic system,and 8 cases involved in both limbic system and non-limbic system simultaneously.22 cases (70.96%)showed low signal in T1WI.25 cases (80.64%)showed high signal in T2WI,with slightly in 15 cases (60%)and obviously in 10 cases (40%).29 cases (93.54%)showed high signal in T2WI-FLAIR,with slightly in 6 cases (20.68%)and obviously in 23 cases (7 9 .3 1 %). The MRI manifestations were not synchronized with the clinical symptoms,the abnormal signal of MRI disappeared later than the clinical symptoms.Conclusion To a certain extent,the MRI findings can provide evidences for diagnosis,therapy,supervision and prognosis of viral encephalitis.

The DNA fragments of ag85b、esat-6、hsp65、mpb64 and ag85b-esat-6、hsp65-esat-6、mpb64-esat-6 were amplified by PCR and SOE technique.These seven fragments were inserted into pCDNA3.1(+)vector to construct recombinant plasmids pCA、pCE6、pCH、pCM、pCAE、pCHE and pCME.The seven plasmids were transfected into SP2/0 cell in vitro to detect the expression of target genes.BALB/c mice were intramuscularly vaccinated with the seven plasmids and the control vector pCDNA3.1(+)and PBS respectively.The serum antibodies and the spleen lymphocyte proliferation(SLP)and secreted IFN～? of spleen were tested.The results of indirect ELISA showed the levels of antibodies in all recombinant plasmids groups were significantly higher than the two control groups(P

Biomarkers ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cysteine Proteinase Inhibitors ; biosynthesis ; genetics ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; analysis ; Liver Neoplasms ; metabolism ; pathology ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; RNA, Messenger ; biosynthesis ; genetics

Objective To investigate the protective effect of dexamethasone on hearing loss induced by canalwall-down mastoidectomy.Methods A total of 76 patients (76 ears) were randomized to dexamethasone group and non dexamethasone group with 38 patients in each group.For dexamethasone group,gelfoam soaked with dexamethasone (5 mg/ml) was topically applied to the round window niche at the end of the surgery,and dexamethasone was administrated intravenously at the dosage of 5 mg/50 kg immediately after the surgery.While for non-dexamethasone group,dexamctbasone was not used.The pure-tone audiograms were performed before mastoidectomy and 90 days after the surgery.Absolute bearing change was defined as the difference in hearing thresholds in decibels before and after the surgery.Results The changes of bone conduction thresholds in dexamethasone group were 8.3± 3.9 dB at 6 kHz,11.3±5.2 dB at 8 kHz,and 10.1±7.2 dB for HPTA (4-6-8 kHz high tone average).As in non-dexamethasone group,the changes of bone conduction thresholds were 13.7±4.6 dB at 6 kHz,25.1±5.4 dB at 8 kHz,19.3±9.7 dB for HPTA.There were significant differences in the changes of bone conduction thresholds between dexamethasone and non dexamethasone groups at frequencies of 6 kHz (P=0.039),8 kHz (P=0.007) and HPTA (P=0.009).Conclusion The results demonstrated that application of dexamethasone significantly reduced sensorineural high frequencies (6 kHz and 8 kHz) hearing loss after canal-wall down mastoidectomy,thus the use of dexamethasone is recommended.