Objective To investigate the effect of PNS on malignant melanoma and the expression of connexin32 in melanoma. Methods The spontaneous and experimental lung metastasis models of B16 melanoma were used to investigate the inhibitory effect of PNS on tumor growth and metastasis. The expression of connexin in melanoma were detected by immunohistoehemistry. Results (1) PNS can obviously inhibit the growth of B16 melanoma. The inhibition rate of the 480mg/kg PNS group was 50. 85%. (2) PNS can obviously inhibit the lung metastasis of B16 melanoma. The number of tumor colonies in lung of the 240 mg/kg PNS group and 480 mg/kg PNS group were lower than that in the negative control group. ( 3 ) The expression of connexin32 in melanoma was detected by immunohistochemistry:PNS could up-regulate the expression of connexin32 in membrane of melanoma. Conclusion PNS can inhibit malignant melanoma growth and metastasis and could also up-regulate the expression of connexin32 in membrane of melanoma.

Objective To investigate the related factors analysis of hepatitis B liver cirrhosis with secondary glucose metabolism disorders.Methods 184 patients with hepatitis B liver cirrhosis were selected as the research object and were given 75g oral glucose tolerance test( OGTI),then they were divided into three groups[ normal glucose tolerance(NGT) group,impaired glucose tolerance and impaired fasting glucose regulation impairment(IGR) group,diabetic( DM ) group ] according the results.The general information,laboratory examination and viral load of three groups were compared.Results ( 1 ) The difference of age,waist size,weight,body mass index data and hepatitis B virus load in IGR group and DM group compared with NGT group were statistically significant difference ( all P ＜0.05 ),IGR compared with DM group,all index comparative differences were not significant (all P ＞ 0.05 ).(2)As the aggravating of glucose metabolism disorder,the positive rate of HBsAg,HBeAg and former S1 antigen of NGT group,IGR group and DM group were gradually raised ( all P ＜ 0.05 ).Conclusion The body mass index and hepatitis B virus load are relatively independent risk factors of secondary glucose metabolism disorders in patients with hepatitis B liver cirrhosis,and which are positive correlation with it.However,gender,age,blood lipids are not obvious correlation with secondary glucose metabolism disorders.

Objective To study the clinicopathology characteristics of chronic lymphocytic thyroiditis (Hashimoto's thymiditis, HT) with occult papillary thyroid carcinoma (occult PTC) in order to improve its diagnostic and therapeutic accuracy. Methods A restrospective analysis was done on the clinical materials of 28 cases of HT with occult PTC from July 1999 to July 2005. All cases were confirmed by operation and pathologic biopsy. Clinical and gross findings were collected. All HE slides were reexamined and immunostains for CK19, galectin-3,and bcl-2 were performed (Envision method). Results In total 189 cases of chronic lymphocytic thyroiditis,28cases (14.8%,28/189)had coexistent occult papillary thyroid carcinoma. There were 19 females and 9 males with median age of 36.7 years old. Normal TSH presented in 11cases (39.3%), high in 8 cases(28.6%), and low in 9cases (32.1%). All tumor diameter was counted for <0.8 cm, 16 cases (57.1%) tumor diameter 0.2～0.5 cm, 12 cases (42.9%) >0.5 cm. Coarse calcification was seen in 6 cases(21.4%) in color ultrasonic exam and CT scans. Follow-up data showed that 28 patients were all alive with no evidence of recurrence or metastasis for 2 to 7 years by December 2007. Conclusion There are no special clinical characteristics in coexistent HT with occult PTC.Coarse calcification in HT in the group of middle-aged women increase the likelihood of the diagnosis. But the diagnosis depends on pathology. Because of the high incidence of occult PTC in HT population, it would be necessary to keep an eye on this particular type of thyroid carcinoma, and multiple sampling in suspected area of HT specimen is advised in the hope not to miss any small tumor in clinical practice.

Objective To examine an in vivo method for the differentiation of induced pluripotent stem cells (iPSCs) into thymic epithelial cells (TECs) in mice. Methods Green fluorescent protein-expressing iPS cells, derived from C57BL/6 mice, were injected into blastocysts from ICR mice. Chimeric blastocysts were then transferred into uteri of E2.5 pseudopregnant mice. Chimeric mouse could be identified by coat color 10 days after birth. The chimeric thymus was transplanted under the renal capsule of BALB/c nude mice. The spleen was cut out from the thymus-transplanted nude mice and the cells were dispersed and analyzed by a flow cytometer 4 weeks after transplantation. Results Chimeras were born 17 days after embryo transfer and 13 live-born chimeras were obtained. The contribution of iPSC-derived cells in the chimeras ranged from 5% to at most 90%. Typical thymic epithelium structure consisted of green fluorescent protein-expressing cells in chimera. The iPSCs-derived thymic epithelial cells could support the generation of new T cells. Conclusion The results indicate that mouse iPS cells can differentiate in vivo towards normally functioning TECs.

Objective To explore the effects of iPS cells-derived chimeric thymus transplantation on T cells reconstitution and graft versus host disease of murine after allo-BMT. Methods iPS cells-derived chimeric thymus was grafted under the renal capsules of mice after allogeneic IBM-BMT. The mice were divided into three groups:IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group. Four weeks after BMT, T lymphocyte subsets in the peripheral blood were analyzed by flow cytometry, the degree and pathological examination of GVHD were observed, respectively. Results Percentage of CD8+T cells in IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group was(5.52 ± 0.83)%,(11.10 ± 1.49)%and(8.49 ± 0.82)%respectively, there was signifi-cant difference between pairwise comparisons(P<0.05), and percentage of CD4 + T cells of the peripheral blood in IBM-BMT+TT group(9.60 ± 0.69)%was significantly higher than IBM-BMT group(6.42 ± 1.40)%and IBM-BMT+DLI group(8.07 ± 0.65)%(P<0.05) . IBM-BMT group and IBM-BMT+TT group showed less clinical and histopathological scoring of GVHD than IBM-BMT + DLI group. Conclusion iPS cells-derived chimeric thymus transplantation could effectively accelerate T cells reconstitution and prevent GVHD after allo-BMT.

Objective To explore the mechanism by which the anti-human P185erbB2 scFv-Fc-IL-2(HFI)modulates tumor surface molecules and activates immune effector cells in vitro. MethodsMTT assay was used to test the proliferation and the LAK-like cytotoxicity. Flow cytometry assay was used to test the expression of ICAM-1, Fas and erbB2 receptors in tumor cells and the expression levels of CD molecules FasL and LFA-1 in human PBMC. Antibody-dependent cell-mediated cytotoxicity(ADCC)mediated by HFI against SKOV3, MCF-7 and SGC-7901 tumor cells was explored hv LDH release assay. Results The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI were upregulated, from 24.85％ and 0.53％ to 85.36％ and 59.19％ respectively, while the expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI were downregulated, from 98.48％, 42.60％ and 36.66％ to 94.01％,30.95％ and 12.36％ respectively. HFI could significantly enhance the proliferation activity of human PBMC, and CD3+ CD8+ T cells and CD3- CD16+ CD56+ NK cells were elevated, from 24.37％ and 6.90％ to 38.80％ and 13.45％ respectively. The expression levels of CD25, LFA-1 and FasL were significantly enhanced from 3.99％, 86.52％ and 5.02％ to 12.96％, 99.06％ and 16.19％. The LAK-like cytotoxicity of human PBMC treated with HFI against SKOV3, MCF-7,SGC-7901 tumor cells was significantly improved:HFI was effective in mediating ADCC against SKOV3,MCF-7 and SGC-7901 tumor cells which expressed high,medium and low levels of erbB2,respectively,and HFI-induced ADCC was correlated with the degrees of erbB2 expression on the tumor cells. Conclusion The expression levels of ICAM-1 and Fas on SKOV3 cell treated with HFI are significantly upregulated. The expression levels of erbB2 on SKOV3, MCF-7 and SGC-7901 tumor cells treated with HFI are downregulated. HFI can significantly enhance the proliferation activity of human PBMC. The LAK-like eytotoxicity of human PBMC treated with HFI against tumor cells is significantly enhanced. HFI iS effective in mediating ADCC and the activity of HFI-induced ADCC is correlated with the degrees of erbB2 expression on the tumor cells.

Objective To investigate the clinical efficacy of Shenmai injection (SMI) combined with heart transplantation of autologous bone marrow stem cells in the treatment of refractory heart failure(RHF). Methods Two hundred patients with RHF were selected from the Emergency Department of the First Affiliated Hospital of Zhengzhou University and the Cardiology Department of the First Affiliated Hospital of Henan College of Traditional Chinese Medicine. They were randomly divided into control group and treatment 1,2 and 3 groups(each 50 cases). In the control group,heart failure standard treatment was given;group 1 received a standard treatment for heart failure combined with autologous bone marrow stem cell heart transplantation;group 2 received a standard treatment for heart failure with SMI;group 3 received a standard treatment for heart failure combined with autologous bone marrow stem cell heart transplantation and SMI. The mean follow-up was 24 months. The prognosis,readmission rate,clinical efficacy,cardiac function and the change in levels of B-type natriuretic peptide(BNP)of patients in each group were observed during the therapeutic course and observation period. Results During the course of treatment,there were 10 cases dead in the control group,4 in each group 1 and 2 respectively,and 3 in group 3. Readmission rates in group 1,2 and 3 were significantly lower than that in control group(38%,36%,24%vs. 48%, P<0.05 or P<0.01). The rates of total efficiency in group 1,2 and 3 were obviously higher than that in the control group(88%,86%,94%vs. 76%,all P<0.05). After treatment,the left ventricular ejection fraction(LVEF),left ventricular fractional shortening(FS)and left ventricular end-systolic diameter(LVESD)in three therapeutic groups were significantly higher than those before treatment,while the left ventricular end-diastolic diameter(LVEDD)and BNP level were significantly lower than those before treatment. All the above indexes in three therapeutic groups after treatment were much more remarkably improved than those in the control group during the same period,and the group 3 being the most significant〔LVEF:0.477±0.099 vs. 0.396±0.098,FS:(30.0±5.1)%vs.(26.8±7.5)%,LVESD (mm):40.6±9.1 vs. 45.8±9.4,LVEDD(mm):44.9±9.8 vs. 52.8±10.1,BNP(ng/L):515±400 vs. 1 875±400, all P<0.05〕. Conclusion SMI combined with heart transplantation of autologous bone marrow stem cells has obvious therapeutic effect for treatment of RHF.

ObjectiveTo observe the lung injury of rats caused by PM2.5 induced imbalance of TH17/Treg immune system and the intervention effect of two different TCM treatments.Methods Wistar male rats were randomly divided into normal group, model group, TCM-treated group1 and TCM-treated group 2. PM2.5-induced lung injury model was established by airway instillation. Model group was given normal saline for gavage. TCM- treated group 1 and 2 were given Yupingfeng Powder and Guomin Decoction combined with Zhisou Powder for gavage. The pathological changes of bronchial and lung tissues, the contents of IL-8, IL-10, IL-17, NE, and MUC5AC in serum and BALF were compared, and the expressions of Foxp3 and IL-17 in lung tissue of each group were analyzed.Results Compared with normal group, the contents of IL-8, IL-17, NE and MUC5AC in serum and BALF of model group increased significantly, while IL-10 decreased significantly (P<0.05,P<0.01); the expression of IL-17 increased significantly and the expression of Foxp3 decreased significantly in lung tissue (P<0.01). Compared with model group, the contents of IL-8, IL-17, and NE decreased in TCM-treated group 1 and 2, while the content of BALF IL-10 increased significantly (P<0.05). The content of IL-10 in serum increased significantly in TCM-treated group 2 (P<0.05); the protein expression of IL-17 of lung issue decreased significantly, and the protein expression of Foxp3 increased significantly (P<0.01). The pathological changes were improved significantly.Conclusion PM2.5 can induce lung injury caused by the imbalance of TH17/Treg. Both two treatments can significantly improve the lung injury induced by PM2.5 and the imbalance of TH17/Tregs immune system.

OBJECTIVE: To explore the expression and significance of Eotaxin and RANTES in the rat model of allergic rhinitis (AR). METHOD: 20 female SD rats in 6-7 weeks were randomly divided into control group and AR group (n = 10, respectively). AR rat model was made with ovalbumin stimulation. To detect pathological changes in mucosa and chemokine Eotaxin, RANTES in their nasal and lung tissues after execution. RESULT: Compared with the control group, Lung EOS cell counted higher in AR group and the difference was significant (P < 0.01); the AR rats nasal mucosa and lung tissue of Eotaxin, RANTES expression was significantly increased (P < 0.01). CONCLUSION There exist high expression of Eotaxin, RANTES, infiltration of eosinophils in nasal and lung tissue of model rats with allergic rhinitis, inferring that the upper and lower respiratory tract inflammatory response has obvious consistency.
Animals ; Chemokine CCL11 ; metabolism ; Chemokine CCL5 ; metabolism ; Disease Models, Animal ; Female ; Lung ; metabolism ; Nasal Mucosa ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic ; metabolism

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.