Objective To study the change of TLR4 on peripheral blood monoeytes (PBMCs) and its role in the pathogenesis of chronic severe hepatitis B. Methods The expression of TLR4 on 10000 CDI4 + PBMCs was determined by flow eytometer in 30 healthy control,31 patients with chronic hepatitis B and 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor α(TNF- α) was determined by ELISA. Results The values of TLR4 on PBMCs and serum TNF-αof the groups of healthy control, patients with chronic hepatitis B and patients with chronic severe hepatitis B were 2.3±1.1,3.7±2.3, (6.9±4.1 ) mean fluorescence intensity (MFI) and (53.8±38.1 ), ( 164.3±89.9) and (359.8±140.0) ng/L. The TLR4 value in patients with chronic severe hepatitis B was signifi- cant higher than those in healthy control and the patients with chronic hepatitis B ( P <0.05). However, there was no significant difference between the patients with chronic hepatitis B and healthy control ( P > O. 05 ). TNF-α increased gradually and significantly from the healthy control to the patients with chronic hepatitis B and patients with chronic severe hepatitis B. There was a significant positive correlation be- tween the value of TLR4 and the value of serum TNF-αin the patients with chronic severe hepatitis B( r=0.666, P <0.01). Conclusion There may be a role of TLR4 in the pathogenesis of chronic severe hepatitis B.

Objective To investigate the distribution of the CTX-M- extended spectrum beta-lactamase (ESBLs) producing Esche- richia coli(ECO) and the molecular mechanism of dissemination. Methods To analyze the drug resistance of the 43 isolates, Kirby-Bauer susceptibility method was used. Multiple polymeraso chain reaction (PCR) was used to amplify the gene of ESBLs, AmpC, full length of blaCTX-M-like gene, insertion sequence (IS) ISEcp1B, IS903 , IS26 and integron I. NEST-PCR was used to detect if the beta-lactamase gene lo- cated in the integron I. The product of full length of bla-CTX-M like gone amplified by PCR was sequenced. Results Susceptibility test showed the resistance from high to low in turn was Ampicillin (97.68%), Coftriaxone (67.44 % ), piperacillin(65.12 % ), Cefotaxime (62.79 % ) ,Coftasidime(58.14% ), Cofasolin(55.81% ), Cofepime (53.49%), Cefexitin(51.16%), ciprofloxacin (44. 19% ), Aztreo- nam(41.86% ), Cefoperasone/Sulbactam ( 20.93% ), Amikacin (0% ), Imipenem (0% ), respectively. ECO was susceptive to Imipenem. CTX-M-G1 was found in 25 strains of ECO , TEM, SHV, CTX-M-G1, ISEcp1B, and integron I were found in the nine isolates. IS903 were found in ECO 3 and 5, and IS26 was found in ECO 3. In ECO 3 and 5, blaCTX-M-like was flanked upstream by ISEcp1B element that provided -35 and -10 promoter sequences and a right inverted repeat (IRR) recognized by transposase, downstream by IS903 provided an inverted re- peat, ISEcp1 B and IS903 composed the complex transpeson. Conclusion ISEcplB may drive the expression and dissemination of blaCTX-M-like gene at a high level.

Objective The person's Mesangial Proliferative Glomemlonephritis (MsPGN) were divided into 3 types based on clini- cal manifestations: nephroticsyndrome, hematuria and proteinuria. To investigate the expression of CD11a, CD11b, CD62L and its signifi- cance in the kidneys patients with MsPGN. Methods CD11a, CD11b and CD62L expression in blood of MsPGN patients (n=35) were investigated by flow eytometry method, and the changes of these proteins in kidney were surveyed. Results In MsPGN, CD11a and CD11b expression in blood were significandy lower and CD62L expression in blood were markedly higher than that in normal humans. And the renal glomeruli of MsPGN also expressed CD11a, CD11b and CD62L. Conclusion The expression of CD11a, CD11b and CD62L are abnormal in MsPGN, and apoptosis may play certain role in the pathogenesis of MsPGN.

Objective To analyze the distribution of T lymphocyte subsets in tumor microenvironment of papillary thyroid carcinoma (PTC). Methods The levels of CD3+ CD4+ CD8- ,CD3+ CD4- CD8+ ,CD3+ CD4+ CD8+ ,CD3+ CD4- CD8- T cell and CD4+/CD8+ ra- tio in tumor microenvironment of 35 cases PTC, 25 cases benign thyroid lesion and 15 healthy eases were detected by flow cytometry. Results Local tumor tissue of 35 cases PTC and 25 cases benign thyroid lesion, and blood of 15 healthy cases were collected. The ratio of CD4+/ CD8+ in blood, tumor microenvironment of benign thyroid lesions and PTCs decreased significantly ( P<0.01). The levels of CD3+ CD4+ CD8+ cell in tumor microenvironment of PTC and benign thyroid lesion were higher than that in peripheral blood( P<0.01). But the differ- ence of the level of CD3+ CD4+ CD8+ cell between PTC and benign thyroid lesion was not significant ( P>0.05). The levels of CD3+ CD4- CD8- cell in tumor microenvironment of benign thyroid lesion were higher than those in PTC and peripheral blood(P<0.01). The difference of the level of CD3+ CD4- CD8- cell between PTC and peripheral blood was not significant (P>0.05 ). The differences of CD4+/CD8+ ratio and the level of each of T lymphocyte subset such as CD3+ CD4+ CD8-, CD3+ CD4- CD8+, CD3+ CD4+ CD8+, CD3+ CD4- CD8- cell between 15 PTC cases with lymph node metastasis and 20 PTC cases without metastasis were not significant (P>0.05). Conclusions CD8+ T cell is the main immune cell in tumor microenvironment in PTC. The local cell-mediated immunity of PTC is in the status of obviously suppressed. But the obvious increase of CD3+ CD4+ CD8+ and CD3+ CD4- CD8- T cells in tumor microenvironment in thyroid benign lesion may be beneficial for creating the location for immune responding against tumor. The distribution of T lymphocyte sub- sets in tumor microenvironment in PTC is not associated with lymph node metastasis.

Objective To investigate the relationship between apoptcsis, expressions of VEGF and clinicopathological characteris- tics, and prognosis in esophageal squamous cell carcinoma (ESCC). Methods Sixty-one surgical specimens of primary esophageal squa- mous cell carcinomas were examined for VEGF by immunohistochemical staining (S-P). Apoptcsis was determined by TUNEL (TdT media- ting dUTP-biotin nick end-labeling) method. Clinicopathologic features were examined by histopathology. The prognostic impacts of these pa- rameters were analyzed by univariate and survival analysis. Results AI and VEGF were well correlated with differentiation, TNM stage. Lower tumor differentiation and higher TNM stage were related to decreasing AI and VEGF. In addition, VEGF in the groups of invasion be- yond muscularis and lymph node metastasis is significant higher than that in invasion reaching muacularis and no lymph node metastasis (P <0.01). But there were no significant correlation between AI and invasion( P>0.05). The simple-factor analysis results showed that the decrease of AI, VEGF, lymph node metastases, lower tumor differentiation, and invasion reaching muscularis were related to decrease of sur- vival rate. However, multivariate Cox analysis demonstrated that only AI and VEGF were the significant prognostic factors. Conclusions Apoptosis and angiagenesis participate in ESCC and promote its growth. VEGF is related to angiogenesis of ESCC. The increase of VEGF may promote invasion and lymph node metastasis. AI and VEGF are significant prognostic factors in ESCC.

Objective To investigate the effect and the mechanism of up-regulating histone acetylizad level with a selective inhibitor of HDACs-Valproate acid sodium (VPA) on breast cancer cell proliferation. Methods MCF-7 cells were cultured with 0.75-4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours in vitro, the inhibiting rate was tested by MTT assay. Cell cycle was analyzed by flow eytome- try with PI assay, and the protein and mRNA expressions of Cyelin A, Cyclin DI, Cyclin E, P21Waf/cipl of MCF-7 cells after 1.5, 3.0 mmol/ L VPA treated were analyzed by indirect immunofluorescence technique and RT-PCR respectively. Results After cultured with 0.75 -4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours, the inhibiting rate of experimental groups increased significantly(P<0.01) and a dose and acting time dependent manner was found. As to cell cycle, the percentages of GI, S, M phrase in control groups remained the same. Contrary to control groups, 0. 75 -4.0 mmo]/L VPA induced a significant arrest in G1 phrase ( P<0.01), and a total of 55.4% -82.8% G1 phrase ratio were found. P21Waf/cipl was up-regulated both at the mRNA and protein level while Cyclin D1 was down-regulated ( P<0.001). Conversely, neither mRNA nor protein expression of Cyclin A, Cyclin E showed difference ( P>0.05). Conclusions Up- regulating histone acetylizad level can inhibit breast cancer cell proliferation, induce cell cycle arrest in G1 phrase. VPA, as a I class of histone deaeetylase inhibitor, can be used as an option in the treatment of breast cancer. The mechanism may include up-regulating P21Waf/cipl mRNA and protein expression and down-regulating Cyclin D1 mRNA and protein expression.

Objective To investigate the role of dynophin and substance P in pruritus with 5/6 subtotal nephrectomy rats (STNx). Methods 5/6 subtotal nephrectomy rots were prepared. After 24 weeks , STNx rats entered in end stage renal disease(ESRD). Sixty male SD rats were divided into 4 groups, STNx+P group(n=15), STNx+dynophin group( n=15), STNx control group(n=15) and sham operation group(n=15). Substance P (SP), dynophin and saline were intradermal injected separately. After injection, scratch reaction of rats in 30 minutes were recorded. And then SP concentration in blood was estimated by ELISA and SP expression in skin was examined by immunohistochemisty. Results SP concentration in blood of STNx rats were (1010.2±103.5)pg/ml, which increased to (2530.0± 236.3) pg/ml in STNx+SP group and decreased to (612.4±72.2)pg/ml in STNx+dynophin group, and it was the lowest in sham opera- tion group (240.2±36.5)pg/ml. SP expression in skin was similar to that of in blood. The scratch times in STNx+SP group were highest (7.3±1.9 times), there was no significant difference between STNx + dynophin group and STNx control group. Conclusion Pruritus in ESRD rats was correlated to the increase of SP in blood and skin. Intradermal injection of dynophin can decrease SP in blood and skin, but can not induce scratch in STNx rats.

Objective To investigate the effect of smad7 on AGEs-induced NRK52E cell tranedifferentiation. Methods NRK52E cells were transferred with Smad7 gene and the cell lines of doxcycllne (Dox) -regulated Smad7 expression were selected for the study. The NRK52E cells were random divided into two groups, Dox - group, the NRK52E cells were cultured with medium only containing AGEs, Dox +group, the NRK52E cells were cultured with medium containing AGEs and Dox. Nuclear translation of p-Smad2/3 were examined with immunocytochemistry. The expression of SmadT, α-SMA, E-cadherin protein were detected by western blot. Results Expression of Smad7 protein in the Dox + group was increased higher than that in Dos- group( t = 3.05, P<0.01). High expression of Smad7 caused a marked inhibition of p-Smad2/3 transnuclear location at 30min and 24h(t=4. 5,t= 3.2, P<0.01). Overexpression of Smad7 dramatically de- creased protein expression of α-SMA, while evidently increased protein of E-cadherin in the NRK52E cells stimulated by AGEs(t=3.47, t=3.25, P<0.01). Conclusion Overexpression of Smad7 can inhibit AGEs - induced transdifferentiation in NRK52E cells.

Objective To study the association between serum total IgE level and β2-adrenoreceptor polymorphism (β2-AR) in asthmatic patients. Methods ELISA was used to determine serum total IgE and AS-PCR was used to determine β2-adrenoreceptor polymor- phism in 44 asthmatic patients. Results In β2-AR 16 locus genotype, the distribution frequencies of Arg/Arg, Gly/Gly showed increasing tendency, whereas Arg/Gly showed decreasing tendency, in normal serum total IgE group and increased serum total IgE group. But there was no significant difference between this two groups. In β2- AR 27 locus genotype, the distribution frequency of Gln/Gln genotype accounts for 76.2% and Gln/Glu genotype for 19.0% in increased serum total IgE group, while Gln/Gln genotype accounts for 9.0% and Gln/Glu geno- type for 73.9% in normal serum total IgE group. There was significant difference between two groups ( P<0.01 ). Conclusions Serum total IgE was correlated with β2-AR 27 locus genetic polymorphism, which may contribute to understand the mechanism of asthma in the peo- ple of the Han nationality in GuiZhou.

Objective To study the expression of Survivin protein and its relation with the expression of Bcl-2, Bax in the tropho- blasts of human normal placenta. Methods The normal placental tissues (8 -9week ,18-23 week and 37-40week) were fixed, embed- ded, sectioned, and Survivin, Bcl-2, and Bax in the trophoblasts were detected with immunnohistochemistry. Result As the gestational age advanced , the staining intensity of Survivin in the trophoblasts was significantly decreased from the first trimester group to the second trimes- ter group to the term group (PU value: 11.74±0.8,9.95±0.43,8. 83 ~ O. 67, respectively, P <0.01 ), while the staining intensity of Bcl-2 and Bax in trophoblast was significantly increased (PU value of Bcl-2 : 4.33±0.60, 5.00±0.75,6.87±0.45, respectively, P<0.01 and PU value of Bax: 9.82±1.12,16.00±1.05,27.48±2.10, respectively, P <0.01 ). Expression of Survivin in trophoblasts has no rela- tionship with the expression of Bcl-2 and Bax (P>0.05 ). Conclusion Survivin may take part in the development of human normal pla- centa through the way of suppressing the apoptesis in trophoblasts. Expression of Survivin in trophoblasts has no relationship with the expres- sion of Bcl-2 and Bax, which indicate that they regulate apoptesis of trophoblasts via different biological pathways.