Objective To assess the application of IS6110-restriction fragment length polymorphism (RFLP)and Spoligotyping in the epidemiology of tuberculosis and to discuss the characteristic of the Mycobacterium tuberculosis in different regions in China.Methods 158 isolates of Mycobacterium tuberculosis strains, were verified by IS6110-RFLP and spoligotyping.Results The discriminatory power of IS6110-RFLP was higher than that of spoligotyping. The result of spoligotyping was compared with the international spoligotype database. Fourteen types belonged to the shared types, in which the type 1 was epidemic and widely,called Beijing genotype. There was significantly difference among the mycobacterium tuberculosis between Guangdong and other regions in clustered rate and the proportion of Beijing genotype (P

Objective To investigate the expression and clinicopathological significance of CXCR4 and HIF-1α protein in esophageal squamous cell carcinoma tissue and explore their correlation.Methods The expression of CXCR4 and HIF-1α protein were assessed by immunohistochemistry SP method in 56 cases with esophageal squamous cell carcinoma and in 20 cases with surrounded normal tissue.Results The expressions of CXCR4 and HIF-lα in esophageal squamous cell carcinoma were significantly higher than those in normal tissues[62.50 ％ (35/56) vs 10.00 ％ (2/20); 57.14 ％ (32/56) vs 0(0/20)](x2=16.259,19.740,P ＜0.01).The expression of CXCR4 and HIF-1α were both correlated with invasion depth (x2 =4.736,7.665,P ＜0.05) and lymph node metastasis ( x2 =7.207,6.389,P ＜0.05),and had no correlation with cancer cell differentiation.The expressions of CXCR4 and HIF-lα in esophageal squamous cell carcinoma were positively correlated (r =0.298,P ＜0.05).Conclusion CXCR4 and HIF-1α are highly expressed in esophageal squamous cell carcinoma.The two have a close relation to esophageal squamous cell carcinoma growth,invasion and metastasis.

OBJECTIVE: Oral zhongfeng an liquid is a new dose form of traditional Chinese medicine for treating cerebrovascular diseases. Its main components are astragalus and hirudo, the former is of obvious effects of replenishing qi and activating blood, and the latter is of stronger effects of antiplatelet, antithrombosis and arteriospasm-reducing, as well as improving tissue anoxia.OBJECTIVE: To investigate the effects of oral zhongfeng an liquid on arterial thrombosis in rats, and blood coagulation and tolerance in mice.DESIGN: A completely randomized and controlled trial.SETTING: Pharmacological Division of Basic Medical College, Hebei Medical University.MATERIALS: The experiment was completed from September 2001 to April 2002 at the Pharmacological Division of Basic Medical College,Hebei Medical University. The experiment of effect of the drug on thrombosis in rats: In the first study, totally 40 Wistar rats were at random divided into five groups: zhongfeng an liquid 3.0, 6.0, 12.0 g/kg, aspirin 0.3 g/kg and control group, with 8 in each group. In the second study, totally 50 Wistar rats were also at random divided into five groups: zhongfeng an liquid 3.0 and 6.0 g/kg, naoxue kang 3.0 and 6.0 g/kg and control group,with 10 in each group. Clotting time study in mice: Totally 50 mice were randomly divided as zhongfeng an liquid 6.0, 12.0, 24.0 g/kg, aspirin 0.3 g/kg and control group, with 10 in each group. Measurement of swimming exhaustion time of mice: Totally 90 mice were randomly divided as zhongfeng an liquid 6.0 and 24.0 g/kg, naoxue kang 6.0 and 24.0 g/kg, benzedrine 0.2 g/kg and control group, with 15 in each group.METHODS: In the experiment of effect of the drug on thrombosis: For the first study, 24 hours and 1 hour before operation the rats in all groups were respectively by gavage given oral zhongfeng an liquid (3, 6, 12 g/kg), aspirin (0.3 g/kg), and water. For the second study, 24 hours and 1 hour before operation the rats were respectively by gavage given oral zhongfeng an liquid (3, 6 g/kg), naoxue kang (3, 6 g/kg) and water. After administration,ketamine 50 mg/kg was peritoneally given for anesthesia, silk ligature thrombosis was used, then the wet thrombus was weighed for comparison of difference in thrombosis among the groups. Measurement of clotting time of mice: The mice were respectively by gavage given oral zhongfeng an liquid (24.0, 12.0, 6.0 g/kg), aspirin (0.3 g/kg) and water, one hour after administration the clotting time of mice was detected with the slide method. Measurement of swimming exhaustion time of mice: The mice were respectively by gavage given oral zhongfeng an liquid (6.0, 24.0 g/kg), naoxue kang (6.0, 24.0 g/kg), Benzedrine (0.2 g/kg) and water, once a day for 5 days.On the fifth day 1 hour after administration, the swimming exhaustion time of mice was measured, the mean value of swimming exhaustion time of mice in each group was calculated.haustion time of mice in each group.RESULTS: All 90 rats and 140 mice involved entered into the result thrombus in rats of the oral zhongfeng an liquid 3.0, 6.0 and 12.0 g/kg groups were obviously lower than those in the naoxue kang groups of the same dose [(24±4), (21±4), (16±6), (39±7) mg, (t=5.88-7.90, P ＜ 0.01)]; in the second study, the wet quality of rats in the oral zhongfeng an liquid 6.0 g/kg group was obviously lower than that in the same dose naoxue kang group [(23.6±2.6), (30.0±4.1), (t=4.18, P ＜ 0.01)], the wet quality of mice in the oral zhongfeng an liquid 3.0 g/kg group was obviously lower than that in the same dose naoxue kang group [(30.6±2.1), (33.1±1.6) mg, (t=2.96,zhongfeng an liquid 24.0 and 12.0 g/kg groups were obviously higher than that in the control group [(222±66), (190±52), (116±26) s, (t=4.02, 4.72, P the oral zhongfeng an liquid 24.0 and 6.0 g/kg groups were obviously higher than that in the control group [(2 512±1 244), (899±403), (502±100) s,(t=3.70-6.24, P ＜ 0.01)].CONCLUSION: Oral zhongfeng an liquid was of obvious inhibition to arterial thrombosis of rats and venous thrombosis of mice, and could enhance the tolerance of mice with a role of antifatigue.

AIM and METHODS: The effects of angiotensin II(AngII) on the activation of three transcription factors were investigated by using EMSA and western-blot methods in endothelial cells respectively. RESULTS: The EMSA results showed that AngII stimulation could increase NF-κB, SP-1 and AP-1 activation in ECV304, which suggested that activity changes in these three transcription factors could partly contribute to the dysfunction of endothelial cells.The binding affinity of NF-κB, SP-1 and AP-1 with corresponding oligonucleotides in AngII-treated ECV-304 were respectively 10.98,3.89,1.33 times as large as in control. The nuclear appearance of AngII-activated NF-κB was examined by western-blot, which corroborates our results from EMSA analysis. While as the protein appearance of AP-1 and SP-1 in nucleus, were little higher than the control group. The result of western-blot suggested that AngII-induced EC activation of these three transcription factors maybe mainly due to the enhanced binding ability to its corresponding cis-acting factors. CONCLUSION: NF-κB, a ubiquitously exposed nuclear transcription factor, is involved, together with SP-1,AP-1, in the regulation of endothelial cell dysfunction related to cardiovascular diseases such as atherosclerosis and hypertension.

AIM: To determine whether the high mobility group protein I (HMGI) is able to bind to the upstream sequence of platelet-derived growth factor B-chain gene and to characterize the HMGI-binding AT-rich regions. METHODS: Recombinant human HMGI (rhHMGI) protein was prepared and electrophoresis mobility shift assay (EMSA) was used. RESULTS: The binding of rhHMGI to PDGF-B (-1 758 / +43 bp) was observed in vitro. Two major HMGI-binding fragments -1 392 / -1 180 bp and -188 / +43 bp were identified, which contained the same AT-rich sequence TTTATAAA (-1 333 / -1 326 bp, -1 314 / -1 307 bp and -30 / -23 bp). An oligonucleotide bound to the TTTATAAA and the GAGACC, the core sequence of the shear stress response element of the PDGF-B, could also bind to the HMGI. Furthermore, HMGI facilitated the binding of NF-?B to the GAGACC in the oligonucleotide. CONCLUSION: The HMGI could bind to the upstream sequence of the PDGF-B gene via the AT-rich sequence TTTATAAA, which may play a role in the transcriptional regulation of the PDGF-B gene.

Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.

Objective: To study the therapeutic value of endoscopic ultrasonography-guided hepaticogastrostomy(EUS-HG) for patients with high malignant biliary obstructive jaundice. Methods: A total of 56 patients with high malignant obstructive jaundice hospitalized at the Second Affiliated Hospital of Nanjing Medical University and the Second Affiliated Hospital of Xuzhou Medical University from January 2014 to December 2017 were included in the study. There were 29 males and 27 females with median age of 72 (60-82) years. Patients were randomized into two groups according to the random number table, the EUS-HG group (n=20) treated with EUS-HG and the percuteneous transhepatic cholangiodrainge(PTCD) group (n=36) treated with PTCD. The operative success rate, curative effect, complications and operation cost were compared between the two groups, and the median unblock period of plastic double pig tail stent was observed. Results: (1)The success rates were 100% in both groups. (2) Preoperative and one-month postoperative levels of the following were tested and compared. Levels of total bilirubin were 362.15±138.27 μmol/L, 56.85±28.57 μmol/L in the EUS-HG group and 356.47±130.69 μmol/L, 60.93±25.79 μmol/L in the PTCD group, respectively. Levels of alkaline phosphatase were 896.57±357.29 U/L, 146.59±48.63 U/L in the EUS-HG group and 883.65±364.32 U/L, 151.57±49.73 U/L in the PTCD group, respectively. Levels of alanine aminotransferase were 252.36±38.77 U/L, 60.29±31.57 U/L in the EUS-HG group and 246.26±32.57 U/L, 62.56±32.87 U/L in the PTCD group. Levels of aspartate aminotransferase were 259.37±30.64 U/L, 62.28±26.58 U/L in the EUS-HG group and 242.37±29.52 U/L, 60.28±29.57 U/L in the PTCD group, and there was no significant difference between the two groups (P>0. 05). CRP levels were 52.57±31.95 mg/L, 16.95±8.77 mg/L in the EUS-HG group and 53.42±35.79 mg/L, 25.13±14.77 mg/L in the PTCD group (P<0.05). (3)There was significant difference in remission rate of anorexia and abdominal distension between the two groups [80.0%(16/20) VS 52.8%(19/36), P<0.05]. There was no significant difference in symptom relief of jaundice, pruritus or abdominal pain between the two groups [90.0%(18 /20)VS 91.7%(33/36), P>0.05]. (4) The incidence of total complications in the EUS-HG group (20.0%, 4/20) was significantly lower than that in the PTCD group (47.2%, 17/36, P<0.05). (5)The cost of operation in the EUS-HG group (22 685.26±2 356.16 yuan) was slightly higher than that in the PTCD group (20 529.57±4 135.63 yuan, P>0.05). (6) The median unblock period of double pig tail plastic stents in EUS-HG group patients was 102 days. Conclusion EUS-HG is a safe and effective method for the treatment of high malignant biliary obstructive jaundice. It can be used as the first choice for treatment after failure of conventional ERCP.