Objective To investigate the relationship of smoking, the gene polymorphisms of heine oxygenase-1, tumor necrosis factor-α, and human β-defensin-1 with chronic obstructive pulmonary disease (COPD). Methods A case-control study was held to compare the genotype frequency distribution of heine oxygenase-1, tumor necrosis factor-α, human β-defensin-1 by PCR -RFLP method and to analyze the relationship among them. Results The L allele gene frequency of heine oxygenase-1 was 23.44% in COPD group and 10.71% in control group with significant difference(P＜0.05). The heterogenesis gene frequency of tumor necrosis factor-α in COPD group and control group were 18. 24% and 7.53% (P＜0.01). The heterogenesis gene frequency of human β-defensin-1 was 14.28% in COPD group and 5.00% in control group with significant difference(P＜0. 01). Conclusions Smoking is a risk factor for COPD. Genetic polymorphisms of heme oxygenase-1, tumor necrosis factor-α, human β-defensin-1 are related with the development of COPD.

Objective To study the characteristics of liver injury induced by simvastatin combined with HRZ (Isoniazid,Rifampicin and Pyrazinamide) in SD rats.Methods Fifty-four 8-week-old SD rats were randomly divided into 3 groups:group A (control),group B (HRZ) and group C (simvastatin combined to HRZ),half of each group were male.We calculated the accurate dose respectively before those rats were given intragastrical administration of corresponding drugs.Six rats were killed in each group on 10th,20th and 40th day,respectively.Before this,blood was fastened from femoral of every rat that would be killed to test liver function,liver tissue slices were made in order to observe the pathological characteristic.Results Alanine amiotransferase of group C elevated in line with time and reached statistic difference on 40th day,furthermore,it was significantly higher than group A (P＜0.05).Total bilirubin and direct Bilirubin of group C were significantly higher than those of group A from the beginning to the end (P＜0.05),however,they declined rapidly on 10th day,this trend also had statistic difference (P＜0.05) At the end of this experiment,hepatic cords was disordered slightly,but swelling liver cells and vacuolar degeneration were observed,the nuleus of cell condensed.Soakage of monocytes,neutrophils,and lymphocytes occurred in the portal and lobule regions,or even spotty necrosis occasionally.Conclusion Cholestasis occurs at the early stage when simvastatin is combined with HRZ in SD rats,however,it has a rapidly degressive trend.In contrast,Alanine amiotransferase elevates,furthermore,pathological injury or even spotty necrosis can emerge in liver tissue slices.

Objective To investigate the effects of the site for access to internal jugular vein (lateral versus anterior),lying position of patients (supine versus Trendelenburg),and head rotation (0°,20°,and maximum) during central venous catheterization on the location depth and cross-sectional area (CSA) of the right internal jugular vein (IJV).Methods Fifteen healthy volunteers were recruited in this prospective observational study from September 2008 to October 2008.Healthy volunteers were placed in flat supine position and 15°.Trendelenburg position separately.In each position,IJV were measured ultrasonographically from lateral site and anterior site with the head oriented at 0°,20°,and maximum rotation separately.Data of measured CSA and location depth of internal jugular vein in different positions were compared.Results The largest CSA (2.16 ±0.89) cm2 and location depth [(1.38 ± 0.43)cm] were occurred at the lateral approach in Trendelenburg position with head oriented at maximum rotation.The CSA in Trendelenburg position was larger than that in flat supine position.Only at the maximum head rotation,lateral approach got statistically larger CSA.The effects of head rotation varied with different degrees of rotation.Conclusions Site of approach,lying position and head rotation had noticeable effects on internal jugular vein cross-sectional area.Trendelenburg position increased the CSA of IJV.

Objective To establish a high throughput screening assay for identifying human small molecular antagonists targeted IL-6R.Methods The full length gene of the human IL-6R extracellular region was amplified by PCR and cloned into a eukaryotic expression vector to construct recombination expression plasmid pABHis -IL6R that was then transfected transiently into HEK293T cells to prepare recombination protein IL-6R.Western blotting assay and receptor-ligand binding experiment were used to analyze the bioactivity of IL-6R.A new screening method based on ELISA was established using the function of IL-6R binding to its ligand and the characteristics of Fc fragment binding to IgG-HRP.Then Z′-factor was calculated and a known antagonist ab 47215 was used to assess the stability and reliability of the new assay .Results Recombination plasmid pABHis-IL6R was constructed and soluble IL-6R was prepared.IL-6R reported herein could be recognized by an anti-IL-6R antibody and specifically bind to its ligand in a dose response manner .A Z′-factor of 0.53 was obtained that could serve high throughput screening assay .Ab47215 , as a known specific antagonist , was able to block rhIL-6 from binding to the receptor in a dose-dependent manner in the new screening assay , the IC50 of which was (0.55 ± 0.11)μg/ml.Conclusion An innovative and easy screening assay for identifying human IL-6R antagonists is established , which might help discover potent and specific antagonists .

Objective To study the influence of carboxymethyl-chitosan(CM-chitosan)on chondro- cyte apoptosis induced by recombinant human interleukin-1?(rhIL-1?)and explore its mechanism.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were pretreated with different concentrations of CM-chitosan for 1 h,then 10 ng/ml IL-1?were added into the culture medium.After 24 h,the apoptotic rates of chondrocytes were measured by flow cytometry with AnnexinⅤ-FITC and PI staining.The morphology of nuclei was observed by fluorescent microscopy with Hoechst 33342 staining.The mitochondrial membrane po- tentials were tested by confocal laser scanning microsocopy with Rhodamine-123 and ATP contents were mea- sured by luciferase reaction.Results CM-chitosan could inhibit chondrocyte apoptosis and restore the func- tion of mitochondria induced by IL-113 in a dose-dependent manner.Conclusion CM-ehitosan can inhibit chondrocyte apoptosis by protecting mitochondrial function.

Objective To evaluate the effect of emulsified isoflurane postconditioning on nuclear factor?E2 related factor 2 ( Nrf2 )?antioxidant response element ( ARE ) signaling pathway during myocardial ischemia?reperfusion ( I∕R ) in rats in vitro. Methods Healthy male Sprague?Dawley rats, aged 4-5 months, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1% amobarbital sodium 40 mg∕kg. Their hearts were excised and perfused in a Langendorff apparatus with K?H solution. Thirty?two isolated rat hearts were randomly divided into 4 groups ( n=8 each ) using a random number table: control group (group C), group I∕R, emulsified isoflurane postconditioning group (EIP group) and fat emulsion group ( group F) . After 20 min of equilibration, group C was continuously perfused with K?H solusion for 100 min. Group I∕R underwent 40 min of ischemia at 32 ℃, followed by reperfusion for 60 min. In EIP and F groups, after undergoing 40 min of global ischemia, the isolated hearts were perfused for 2 min with K?H solution containing 1.68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then were continuously perfused with K?H solution containing oxygen at 37 ℃ for 58 min. Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end?diastolic pressure ( LVEDP ) , and positive maximal pressure of left ventricular increase (+dp∕dtmax ) were recorded at the end of equilibration and reperfusion. At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase?1 ( HO?1) , quinone oxidoreductase 1 ( NQO1) , and superoxide dismutase 1 ( SOD1) and mRNA expression using Western blot and real?time PCR. Results Compared with group C, HR, +dp∕dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I∕R and F groups, LVDP was significantly decreased, LVEDP was increased, and no significant changes were found in HR and +dp∕dtmax at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated at the end of reperfusion in I∕R, EIP and F groups. Compared with group I∕R, HR, +dp∕dtmax and LVDP were significantly increased, and LVEDP was decreased at the end of reperfusion in EIP and F groups, Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was significantly up?regulated at the end of reperfusion in EIP group, and Nrf2, HO?1, NQO1 and SOD1 mRNA expression was significantly up?regulated, Nrf2 and HO?1 expression was up?regulated, and no significant changes were found in NQO1 and SOD1 expression at the end of reperfusion in group F. Compared with group EIP, HR, +dp∕dtmax and LVDP were significantly decreased, LVEDP was increased, and Nrf2, HO?1, NQO1 and SOD1 and mRNA expression was down?regulated in group F. Conclusion Emulsified isoflurane postconditioning attenuates myocardial I∕R injury probably by activating Nrf2?ARE signaling pathway in isolated rat hearts.

OBJECTIVE: To analyze the research literatures related to bioartificial liver, and to make a conclusion concerning the development of bio-artificial liver.DATA SOURCES: Using bioartificial liver, liver cell, hepatocyte culture and bioreactor as search terms, searching Ovid, Springer Link database, and China National Knowledge Infrastructure, Vip Information database and Wanfang Date (1990.09-2008.09). Literatures search was limited to English and Chinese languages.DATA SELECTION: Researches regarding liver cells of bioartificial liver, reactors and auxiliary equipment was included, and the studies about immune and animal infection studies of bioartificial liver were excluded.MAIN OUTCOME MEASURES: ①The source, quantity and culturing of bio-artificial liver hepatocytes. ②Bioreactor type, nature and type of films. ③Composition of oxygen and temperature control devices of bioartificial liver.RESULTS: Totally 3898 documents seized initially in the searching by computer, according to inclusion and exclusion criteria, 29 were analyzed. Bioartificial liver was a hybrid device in which can culture hepatocytes in vitro, when the patient's blood flows through the device, material exchange with the cultured hepatocytes through semi-permeable membrane or direct contacting can take place, which can perform the same roles of detoxification, synthesis, biological transformation and other functions as real liver cells, so as to achieve the purpose of support and treatment. Bioartificial liver can also be involved in metabolism of the three major nutritive substances, as well as secretion of hepatocyte growth promo ting substances. So it is an effective alternative to the real liver as the function of detoxification and synthesis, and can fills the essential gap between the transplantation and acute liver failure.CONCLUSION: Although the bioartificial liver research has made significant progress, it still faces the problems such as limited liver cells sources, long-term maintenance of liver cell activity and function, and further optimization of the reactor design.

Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells (PSCs). Methods Rats' PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro.The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode (LED) with an irradiation energy of 0.5 J/cm2,1 J/cm2,2 J/cm2 or 4 J/cm2.Any effect was confirmed by Alcian blue staining,immunohistochemistry and observing histomorphological changes under a light microscope,as well as detection using a reverse transcription polymerase chain reaction (RT-PCR). Results After being induced for 14 d,the PSCs exhibited polygonal and round shapes. Alcian blue and type Ⅱ collagen immunohistoehemistry staining showed positive results,but the control group had no significant change.RT-PCR showed that the mRNA expression of Sox9 and type Ⅱ collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.

Objective To analyze and compare the mutations in two different regions of the katG gene and study the relevance of Mycobacterium tuberculosis isoniazid-resistance and mutations in two different regions of the katG gene. Methods Fifty-three INH-resistant Mycobacterium tuberculosis strains isolated in cultures of sputum samples obtained from Zhejiang province were analyzed. PCR was used to amplify two regions of the katG gene (GenBank accession no. U06258) region 1 (from codon 1 to codon 150) and region 2 ( from codon 227 to codon 470) which were then sequenced in order to identify mutations. Results Three strains resistant to INH did not contain mutations in either region. Fourteen strains carried mutations in region 1. Among them 5 strains barbered deletions, and showed high-level resistance to isoniazid. Five strains had mutations only in region 1. Region 2 carried multiple point mutations, especially at codon 315, and there were S315 N ( AGC→AAC ) substitution in 18 of those cases. The frequency of mutations in the katG S315 of high-level INH-resistance isolates ( 84. 4%, 27/32) was significantly higher than those of low-level INH-resistance isolates( 15.6%, 5/32 ), there was statistically significant difference (x2 = 30. 25, P ＜ 0. 01 ).katG S315 mutations in high-level INH-resistance frequency (84. 4%, 27/32) was significantly higher than the other mutations of katG gene of high-level INH-resistance frequency (27. 7%, 5/18 ), there was significant difference (x2 = 16.02, P ＜ 0. 01 ). The analysis of region 2 allowed INH resistance to be diagnosed in 84. 9% of the strains. Five strains had mutations only in region 1 ,which allowed the proportion of INH-resistant strains identified to be increased to 94. 3%. Conclusions The number of mutations at codon 315 was high. Mutation type and location closely related with drug resistance and the analysis of region 1 resulted in a 9. 4% increase in the rate at which mutations were identified.

Objective To investigate the effect of HbA1c meeting the standard or not on microalbuminuria,blood lipids and liver enzymes in patients with type 2 diabetes.Methods A retrospective analysis was performed on 457 subjects who had type 2 diabetes.They were divided into substandard group and standard group according to HbA1c result.The general information and relevant laboratory indicators of patients were.collected and compared between two groups.Results The microalbuminuria,serum triglyceride and liver enzymes (glutamyl transpeptidase,alkaline phosphatase,aspertate aminotransferase) were significantly different between two groups [ (189.8 ± 235.3) mg/dl vs (38.9 ± 85.5) mg/dl,(2.64 ± 2.99) mmol/L vs (2.02 ± 1.50)mmol/L,(41.7 ±52.9)U/L vs (29.7 ±24.9)U/L,(83.6 ±28.6) U/L vs (74.3 ±25.8)U/L,(26.7 ±19.1)U/L vs (22.0 ±10.5) U/L,P ＜0.05].HbA1c level was positively correlated with microalbuminuria,glutamyl transpeptidase and alkaline phosphatase (r =0.209,0.115,0.11,P ＜0.01).The microalbuminuria was an independent risk factor of affecting HbA1c to reach the standard (OR = 1.009,P ＜0.05).Conclusions HbA1c meeting the standard or not can influence many factors except blood glucose.