Objective:To study the effect of sulforaphane(SFN) on the growth of human bladder cancer cell and its mechanism in vitro. Method:Morphological characteristics of T24 cell nucleus induced by SFN were observed by AO/EB fluorescein staining .The effect of SFN on the T24 cell cycle was determined by flow cytometry. The expression of TrxR at the transcriptional and translational levels were measured by RT-PCR and Western blot methods respectively. Results:(1) After the cells were treated with 10 ?mol/L and 20 ?mol/L SFN for 24 h and 48 h, nuclear fragmentation and apoptotic body were seen under fluorescence microscope. (2) SFN could block the cell cycle at the G0/G1 phase showed by flow cytometry. Moreover, the treatment of 20 ?mol/L SFN for 48h could cause the appearance of sub-G1 before G0/G1 phase. (3) The expression of TrxR mRNA were increased by the treatment of 10 ?mol/L SFN for 4 h, 10 h,24 h ,compared with the control group. Furthermore ,the treatment with high dose SFN (20 ?mol/L ) for 10h or 24 h could induce the expression of TrxR mRNA more significantly . (4)The expression of TrxR protein in the 10 ?mol/L SFN for 24 h group was augmented compared with the control group , and aftertreatment with 20 ?mol/L SFN for 8 h and 24 h, its expression was significantly higher than that in the control group . Conclusion:SFN can inhibit the growth of T24 cell ,induce apoptosis and arrest T24 cell in G0/G1 phase. Its mechanism is associated with the induction of TrxR both at the transcriptional and translational levels.

Dendritic cells act as the major antigen presenting cells in the body and play a central role in intri-guing the adaptive immune response. Protective immunity against schistosome and immuno-pathological response in host caused by eggs are both closely associated with Th2 response. Further understanding on immune mechanism will contribute to the development of vaccines against schistosome infection, as well as the relief of the pathological lesion in schistosomiasis. This article discusses the central role of dendritic cells in the mechanism of Th2 response induced by schistosome (including eggs).

MicroRNAs (miRNAs) are small noncoding regulatory RNAs ranging in size from 17 to 25 nucleotides which participate many physiological and pathological processes.MicroRNA could also stably exist in peripheral blood,and the detection of circulating microRNA is of great significance for disease diagnosis and prognosis.As so far,there is no unified method and standard for detection of microRNA,which is the major reason for discrepant results between different studies.However,circulating microRNA,as a new disease detection biomarker,has stable properties,convenient detection and high accuracy features,so it till has an important potential value in clinical applications.

BACKGROUND:Whether transplanted bone marrow mesenchymal stem cells under hypoxic conditions can survive is crucial for the successful celltransplantation. Therefore, studies on the growth of bone marrow mesenchymal stem cells under hypoxic conditions in vitro can provide experimental evidence for in vivo celltransplantation. OBJECTIVE:To observe the expression of vascular endothelial growth factor in bone marrow mesenchymal stem cells under hypoxia. METHODS:Rat bone marrow mesenchymal stem cells were obtained and cultured, and observed under light microscopy. Passage 3 cells were cultured under normoxia (21%O2) and hypoxia (3%O2 hours. Then cellcounting kit-8 assay and flow cytometry were employed to detect cellproliferation in the two groups. Western blot assay was adopted to detect the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the two groups. RESULTS AND CONCLUSION:(1)Rat bone marrow mesenchymal stem cells were obtained and cultured successful y, which were fusiform cells and had uniform shape under the light microscope. (2)The results of cellcounting kit-8 assay showed that the number of cells in the hypoxic group was higher than that in the normoxic group at each time point, and cellviability increased significantly at hours 36 and 48 (P<0.05). (3)The results of flow cytometry demonstrated that the proportion of cells in S phase and cellproliferation index in the hypoxic group were significantly increased, compared with the normoxic group (P<0.05). (4)Western blot results showed ), respectively, for 72 that there was a smal amount of the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the normoxic group, but the expression of these two proteins in the hypoxic group was increased in a time-dependent manner (P<0.05). These findings suggest that hypoxia can induce proliferation of rat bone marrow mesenchymal stem cells cultured in vitro, and also raise hypoxia-inducible factor-1αand vascular endothelial growth factor expression in a time-dependent manner.

Objective:To explore the correlation between serum level of matrix metalloproteinase 2 (MMP-2)and se-verity of coronary artery disease in aged patients with acute coronary syndrome (ACS)complicated hyperhomocys-teinemia (HHCY).Methods:According to serum level of homocysteine (Hcy),a total of 105 aged ACS patients were selected and divided into ACS+HHCY group (n=56)and pure ACS patients (n=49),another 65 healthy in-dividuals were enrolled as healthy control group.The severity of coronary artery disease was showed by Gensini score ;enzyme linked immunosorbent assay was used to measure serum MMP-2 level in each group.Results:Com-pared with healthy control group,there were significant rise in MMP-2 level [(140.8±50.1)ng/ml vs.(442.5± 98.2)ng/ml,(297.9±86.3)ng/ml]in ACS+HHCY group and pure ACS group,P <0.01. Gensini score in ACS+HHCY group was significantly higher than that of pure ACS group [(1.9±0.2)scores vs.(1.1±0.3)scores,P<0.01].Pearson correlation analysis indicated that Gensini score was positively correlated with MMP-2 level in ACS+HHCY group (r =0.424,P <0.05).Conclusion:The serum MMP-2 level is significantly positively correla-ted with severity of coronary artery disease in aged patients with ACS complicated HHCY.

Objective:To analyze the effect of tanreqing injection on the serum levels of transforming growth factor-β (TGF-β) and matrix metalloproteinases-9 (MMP-9) of patients with chronic obstructive pulmonary disease (COPD).Methods:102 patients with COPD were divided into the control group and the observation group according to random number table method,52 cases in each group.The control group was treated with routine therapy,and the observation group was treated with Tanreqing injection based on the control group.The serum TGF-β,MMP-9 levels,forced vital capacity (FVC),1 s forced expiratory volume (FEV1),partial pressure of carbon dioxide (PaCO2),oxygen partial pressure (PaO2),CD4+,CD86,CD4+/CD8+,syndrome integral,clinical efficacy and incidence of side effects were observed and compared between the two groups.Results:After treatment,the serum TGF-β,MMP-9 levels,PaCO2 and syndrome integral of observation group were significantly lower than those of the control group,the PaO2,CD4+,FVC,FEV1,CD4+/CD8+ and the clinical efficacy of observation group were obviously higher than those of the control group (P＜0.05).There was no significant difference in the incidence of side effects between the two groups (P＞0.05).Conclusion:Tanreqing injection could effectively reduce the serum levels of TGF-β and MMP-9,and improve the arterial blood gas,lung function and immune function in treatment of patients with COPD.

Objective To compare the effects of propofol versus isoflurane on brainstem auditory evoked potential (BAEP) and explore the difference in the effects of the two anesthetics on the brainstem. Methods Thirty ASA Ⅰ or Ⅱ patients aged 20-50 yr without heating disorder, scheduled for elective surgery performed under general anesthesia were randomly divided into 2 groups (n = 15 each): propofol group (group P) and isoflurane group (group Ⅰ). SpO2, PET CO2, BIS and BAEP were continuously monitored before and during anesthesia. Anesthesia was induced by propofol administered by TCI or isoflurane inhalation. Tracheal intubation was facilitated with vecuronium. The patients were mechanically ventilated. PET CO2 was maintained at 35-40 mm Hg and SpO2 at 98%-100%. After intubation BIS was maintained at 70 and 50 respectively ,the latency of the wave Ⅰ , Ⅱ and Ⅴ and the inter-peak latency (IPL) betwecn wave Ⅰ -Ⅲ , Ⅲ-Ⅴ and Ⅰ -Ⅴ were recorded.Results In group P there was no significant difference in the latency of the wave Ⅰ , Ⅲ and Ⅴ and the IPL between wave Ⅰ - Ⅲ , Ⅲ - Ⅴ and Ⅰ - Ⅴ between the baseline before anesthesia and at BIS 70 and 50. In group Ⅰ the latency of wave Ⅲ and Ⅴ and the IPL between wave Ⅰ - Ⅲ , Ⅲ - Ⅴ and Ⅰ - Ⅴ were significantly longer at BIS 50 than the baseline before anesthesia, while the latency of wave Ⅲ and Ⅴ and the IPL between wave Ⅰ -Ⅲ andⅠ -Ⅴ at BIS 50 were significantly longer than that at BIS 70. At BIS 50 the latency of wave Ⅴ and the IPL between wave Ⅰ -Ⅴ were significantly longer in group Ⅰ than in group P. Conclusion At comparable depth of anesthesia propofol exerts less depressant effects on BAEP indicating less depression of brainstem.

Atomic absorption spectrometry was employed to determine the levels of 7 essential trace elements in the whole blood of 150 mainteinance hemodialysis (MHD) patients.And comparisons were made with the normal group of 200 participants in routine medical examination at the same time at our hospital.And among 7 indices,as compared with the control group,the blood levels of zinc,chromium,cobalt and manganese were significantly lower (P ＜ 0.05) while those of selenium and copper higher (P ＜ 0.05).MHD patients lacked essential trace elements to a varying extent.A clinician should pay attention to complementing essential trace elements to improve the patient's quality of life.

Objective To clone and express the partial encoding sequence of Mr 70 000 heat shock protein of Cryptosporidium andersoni (CaHSP70) in Escherichia coli and identify the recombinant protein. Methods Total RNA was extracted from oocysts of C.andersoni isolated from Xuzhou, Jiangsu (XZ-BOV). The CaHSP70 gene was amplified by RT-PCR. The PCR product was cloned and then subcloned into pET28a vector, and the recombinant plasmids were transformed into E.coli BL21(DE3) subsequently. The expressed protein induced by IPTG was purified and identified by SDS-PAGE and Western blotting, and was further analyzed by relevant bioinformatics softwares. The specific IgG antibodies in mice immunized by rCaHSP70 were detected by Western blotting and ELISA respectively. Results The deduced amino acid sequence showed to be identical with that of C. andersoni Mr 70 000 heat shock protein (HSP70). The recombinant protein expressed in the form of inclusion body was about Mr 43 000. It could be recognized by anti-His G labeled HRP antibodies and all the sera from mice infected with C. andersoni and children infected with C. parvum as well as sera from mice immunized with rCaHSP70 respectively. The rCaHSP70 possibly had multiple domains and potential antigenic determinants. Phylogenetic analysis showed that XZ-BOV and C. andersoni were in the same clade. ELISA showed that the level of specific antibodies against rCaHSP70 in immunized BALB/c and C57BL/6 mice was significantly higher than that of mice before immunization. Conclusion The recombinant plasmid pET28a-CaHSP70 has been constructed. The purified rCaHSP70 exhibits high antigenicity and seems a potential candidate antigen for immunodiagnosis of cryptosporidiosis.

Objective To study the structural features and characteristics of a novel gene Schistosoma japonicum 79(Sj79), and observe its effect of RNA interference(RNAi),so as to provide the experimental basis for its further function study and mechanism study of anti reproductive development of schistosome. Methods The gene structure and characteristics of Sj79 were analyzed by bioinformatics methods. Then the expressions of Sj79 messenger RNA(mRNA)during the different develop?mental stages of schistosome were analyzed and the effects of RNAi silencing were observed by the soaking method. The tran?scriptional levels of Sj79 after RNAi were detected by real time PCR. Results The open reading frame of Sj79 contained 696 base pairs with an exon structure. The gene had obvious stage specificity,and its transcriptional level in mature female worms was the highest. After soaking for 3 d,the Sj79 mRNA level[(41.0 ± 12.3)%]in the siRNA?1 group with low dosage(20 nmol/L) was lower than that in the siRNA?NC group[(103.2 ± 14.4)%],the difference was statistically significant(t=3.28,P<0.05). When with high dosage(200 nmol/L ),both the Sj79 mRNA levels in the siRNA?1 group[(15.8 ± 10.9)%]and siRNA?2 group [(11.1 ± 8.8)%]were significantly lower than that in the siRNA?NC group[(100.1 ± 6.3)%](t=13.44,27.84,both P<0.01). After soaking for 7 d,only the Sj79 mRNA levels in the siRNA?1group[(43.4 ± 4.5)%]and siRNA?2 group[(62.5 ± 5.4)%]with low dosage were lower than that in the siRNA?NC group[(100.4 ± 5.2)%],and the differences had statistical sig?nificance(t=8.33,5.07,both P<0.01). Conclusion Through this study,we have improved the mRNA sequence and genom?ic information of Sj79 gene,and understood its structural features,as well as selected out two effect fragments siRNA?1and siR? NA?2 which will provide the basic evidences for the further study on egg laying interference of the female adult worm of schisto?some in vitro.