AIM: To investigate the effect of combination of traditional Chinese and western medicine treatment on active tuberculosis patients and the impact on peripheral blood T-lymphocyte subsets. METHODS: 133 cases of active pulmonary tuberculosis patients were divided into two groups.The clinical effects of two groups were compared and Tlymphocyte subsets were detected. RESULTS: The total effective rate was 95.8%,its efficacy was superior to the control group(P

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

This article discusses the production of CAI courseware and the problems of its application to teaching process,to make full use of it and acquire satisfactory teaching effects.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

Aim To examine the effect of different dosages hydroxyethyl starch(HES)130/0.4 on intercellular adhesion molecule 1(ICAM-1)expression in lung tissue of acute lung injury in endotoxemic rats and explore the role of MAPKs pathway in its expression.Methods Thirty six healthy Sprague Dawley(SD)rats weighing 270~320 g were randomly divided into 6 groups with 6 animals in each group.In group H1-H4,1 min after lipopolysaccharide(LPS)5 mg?kg-1 intravenously administration,HES 130/0.4 with 3.75,7.5,15,30 ml?kg-1 were infused intravenously respectively at a rate of 0.2 ml?min-1.In group L,saline instead of HES 130/0.4 was administered.Group N served as control by giving the same volume of saline.The animals were anesthetized with pentobarbital.Right external jugular vein was injected with LPS.The animals were killed 4 hours after LPS injection for determination of total protein,WBC,MPO,W/D,ICAM-1 protein and mRNA and MAPKs activity.Results Compared with control group,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were increased significantly in group L.Compared with group L,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were decreased significantly in group H1 and H2,especially at the dosage of 7.5 ml?kg-1.Conclusion HES 130/0.4(7.5 ml?kg-1)can attenuate inflammatory response of acute lung injury induced by LPS,which may be related with inhibiting the expression of ICAM-1 protein and mRNA through MAPKs signal pathway.

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.

Objective To study the association of MIF-173 locus polymorphism and the risk of prostate cancer (PCa) in China. Methods A case control study including 259 PCa patients and 301 age-matched controls was conducted. The polymorphisms of MIF-173 locus were analyzed by poly-merase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique using genomic DNA isolated from peripheral blood lymphocytes. The correlations between the susceptibility to PCa and different genotypes were compared. The effect of age, smoking method and family history of canc-er were also analyzed. Results The rate of the MIF-173 * C variant allele of the PCa patients(n=259) was significantly higher than that of the controls (n=301) (36.0% vs 15.0%). The MIF-173 *C variant allele could significantly increase the risk of PCa (OR=2.96,95%CI: 1.92-4.57). Peo-ple with older age (age>70) or family history of cancer, who carried MIF-173 * C allele demonstra-ted a significantly increased risk in comparison with those carrying wild genotype of G/G(OR=3.66, 95%CI=2.02-6.62;OR=3.26, 95%CI=1.24-8.55). Conclusion These results suggested that polymorphisms of MIF-173 locus appear to influence the risk of PCa and may have synergistic effect with age and family history of cancer.

Objective To evaluate the significance of human papillomavirus L1 capsid protein (HPVL1) and human telomerase RNA component (hTERC) gene in the cytologic specimens of cervix which was infected by the high-risk types of human papillomavirus (HR-HPV),and to expose their relationship with cervical lesions.Methods The fluorescence signal of cytologic samples of cervix were detected by interphase FISH in chromosome enumeration double-color DNA probes TERC.The expression of HPVL1 capsid protein was detected by MaxVision immunohistochemistry method.300 samples were analyzed with HR-HPV positive from the cervical biopsy.The diagnoses as normal or chronic inflammation (n =45),cervical intraepithelial lesions Ⅰ grade (CIN Ⅰ,n =95),CIN Ⅱ (n =58),CIN Ⅲ (n =64),and squamous cervical cancer (SCC,n =37).Results The percentage of HPVL1 positive rates in normal or chronic inflammation,CIN Ⅰ,CIN Ⅱ,CIN Ⅲ and SCC groups were 58.70 ％ (27/46),63.16 ％ (60/95),37.93 ％ (22/58),10.94 ％ (7/64) and 0 (0/37),respectively.The percentage of HPVL1 decreased along with the increase of severity of the cervical intraepithelial lesions.Genomic amplification of hTERC positive rates in normal or chronic inflammation,CIN Ⅰ,CIN Ⅱ,CIN Ⅲ and SCC groups were 6.52 ％ (3/46),11.58 ％ (11/95),51.72 ％ (30/58),85.94 ％ (55/64) and 100.00 ％ (37/37),respectively.The percentage of hTERC increased along with the severity of the cervical intraepithelial lesions (rs =0.302,P ＜ 0.01).The percentage of HPVL+/hTERC-was 57.89 ％ in CIN Ⅰ group and 4.69 ％ in CIN Ⅲ group.The percentage of HPVL-/hTERC+ was 6.32 ％ in CIN Ⅰ group and 79.69 ％ in CIN Ⅲ group.Conclusion The detection of HPVL1 and hTERC are important for assisting cervical lesions screening and monitoring of disease progression in the HR-HPV positive cytologic specimens.