Objective To observe the immunoregulatory roles of MSCs in inducing proliferation of B regulatory cells (Bregs).Method DA Rat MSCs were co-cultured with Wistar rat B cells.The levels of IL-10 and secrete proinflammatory cytokines in the supematants were determined by using ELISA.The proliferation and apoptosis of B cells after interaction with MSCs were analyzed by using FACS.In order to investigate whether Bregs are involved in the phagocytosis of immune complex (IC),B cells and Bregs were incubated with FITC-OVA-IC.The fluorescence intensity was detected by using immunofluorescence microscopy.Results MSCs induced B cells to differentiate into Bregs with high CD19,CD1d and CD5 expression and high IL-10 level.The proliferation of B cells was significantly suppressed after interaction with MSCs.Bregs had enhanced phagocytic capacity as compared with that of B cells.Bregs significantly inhibited maDC-induced CD4+ T cell proliferation compared to B cells,without depressing IL-2 or IFN-γ secretion.Conclusion MSCs induce freshly isolated B cells to differentiate into Bregs characterized by the ability to preferentially produce IL-10.

Objective: To investigate whether the plasmid of short hairpin RNA targeting human insulin like growth factor receptor 1 (pSUPER-siRNA-IGF1R) can enhance mitomycin-induced apoptosis of human liver cancer SMMC7721 cells and the corresponding mechanism. Methods: The siRNA targeting IGF1R was designed and the pSUPER-siRNA-IGFI R plasmid was constructed and transfected into SMMC7721 cells. The stable cell lines were screened by G418. The growth curve of every group was detected by CCK8 method. After mitomycin treatment, the sensitivity of hepatoma cells to chemotherapy was monitored by CCK8 method. The apoptotic index (AI) was examined by flow cytometry. The expression of apoptosis-related proteins p53, Bax, bcl-2 was determined by Western blotting. The empty control group and positive control were set up. Results: The apoptosis induced by mitomycin were significantly enhanced in stable cell line and the AI was significantly elevated compared with other groups (P < 0.05). The expression of wild type p53 and Bax protein was significantly up-regulated but the expression of bcl-2 protein was significantly down-regulated (P < 0.05). Conclusion: The constructed pSUPER-siRNA-IGF1R plasmid silences RNA transcription and enhances the apoptosis of tumor cells induced by mitomycin.

Objective To explore the effects of lactational maternal stress on neurobehavioral development of offsprings.Methods Maternal mice were divided into normal control,maternal deprivation and maternal stress group.A 5-min/day cold water swimming was used as maternal stress for continuously of 14 day since the 7th postnatal day.The increasing bodyweight and neurobehavior of adolescent offspring mice were assessed by forced swimming test,tail suspension test and locomotor activity in the 35th postnatal day.Results Maternal chronic stress in lactational stage significantly increased the vulnerability of offspring to acute stress,which was reflected by significantly increased immobility time of adolescent offspring in forced swimming test (female:(139±6) s,(138±9) s,(96±9) s respectively; male:(139±9) s,(112±9) s,(96±9) s respectively) and in tail suspension test (femal:(127±8) s,(123±8) s,(98±6) s respectively ; male:(141 ±7) s,(105±5) s,(92±6) s respectively).Meanwhile,sexual difference occurred for adolescent offspring to acute stress,and the female offspring were more vulnerable to acute stress.Conclusion Lactational maternal stress may be important factors for development of adolescent depression.Our findings highlight the area of early prevention and intervention for adolescent depression.

[Summary] Malignant obstructive jaundice is the bile obstruction caused by the invasion of cholangiocarcinoma , pancreatic cancer or ampulla cancer .Due to lack of effective treatment , the prognosis is poor .In recent years , with the rapid development of medical technology and imaging technology , dual interventional treatment technology , such as percutaneous transhepatic cholangial drainage ( PTCD) or biliary stenting combined with radioactive seed implantation , ablation catheter lumen combined with biliary stent implantation , is applied in the treatment of malignant obstructive jaundice .This article is the summary of the clinical application of PTCD combined with radioactive seed implantation , biliary stent combined with radioactive seed implantation and intraluminal catheter radiofrequency ablation combined with biliary stent implantation technique .

Objective To study the effect of Fufangbiejiaruanganpian (FFBJRGP) on deposition of collagens around hepatic Disse space in experimental alcohol liver fibrosis (ALF). Methods The ALF rat model was induced by feeding a mixture of alcoholic liquid which included alcohol, vegetable oil and pyrazole, combined with a high fatty food. Then, all of the ALF rats were treated with FFBJRGP in high, moderate, and low dose. At the time of termination of the treatment, HYP level in liver tissue, collagen Ⅳ and LN levels in the serum and pathological changes were studied by Mallory stain and sirius red stain. Results The levels of HYP in hepatic tissues, collagen Ⅳ and LN in sera are increased remarkably in ALF rats, while all these indexes were reduced after moderate or low-dose FFBJRGP treatment (P

Purpose:To investigate the expression of TGF-?1,Cyclin E in (primary) gallbladder carcinoma and its clinical significance. Methods:The expression of TGF-?1,Cyclin E in gallbladder carcinoma was detected by S-P immunohistochemical staining,20 cases of chronic cholecystitis were collected as control. Results:①The positive rate of TGF-?1(63.89%),Cyclin E(47.22%) in gallbladder carcinoma increased significantly(P

Objective To investigate the expression of tumor growth tactor ?1 (TGF-?1) and p27 in gallbladder carcinoma and their relation to the development of the carcinoma. Methods The expression of TGF-?1 and p27 in 36 cases of gallbladder carcinoma was detected by SP immunohistochemical staining. Twenty cases of chronic cholecystitis were collected as control. Results The positive rate of TGF-?1 (63.9%) was higher than that of the control (10.0%), P

Objective To establish hepatocellular carcinoma(HCC) cell lines which olig-expressed IGF1R gene stably.Methods An eukaryotic expressing vector pSUPER-IGF1R-siRNA that could block IGF1R expressing was transferred into hepatocellular carcinoma cell lines SMMC7721 and Hep3B with Lipofectamine 2000 reagents.After transferred,cells were selected with G418 to obtain positive clones.The expressions of IGF1R,cyclin D1 and cyclin B1 were detected by RT-PCR and Western-blot.Cell growth curve were painted.Results Two cell lines clones were screened olig-expressing IGF1R gene stably.The experimental cell lines grew more slowly than control cell lines and the expression of cyclin D1 decreased(P

Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.

Objective To explore the role of integrin αvβ6 in mediating the tolerance of gastric cancer AGS cells to 5-fluorouracil, and to determine whether direct β6-extracellular signal-regulated protein kinase(ERK) binding is involved. Methods Gastric cancer AGS cells in the logarithmic phase were incubated with either 5-fluorouracil for 24 hours ( control group), with 0.1 g/L of mouse anti-αvβ6 monoclonal antibody 10D5 for 6 hours and then with 5-fluorouracil for24 hours ( 10D5 group), with IgG2a and 5-fluorouracil ( IgG2a group), or with 5-fluorouracil and 20 μnol/L of ERK inhibitor PD98059 for 24 hours ( PD98059 group). Cell proliferation,apoptosis and the expression of Bcl-2 and caspase-3 protein were detected by MTT assay, flow cytometry and western blotting, respectively. All data were analysed by one-way analysis of variance and LSD test. Results The cell inhibition rates of the control group, 10D5 group, IgG2a group and PD98059 group were 28.1% ±2.7%,84.5% ± 1.6%, 31.4% ±5.2%, 86.7% ±5.2%, respectively, with a significant difference ( F = 342. 5, P ＜0.05). The apoptosis rates of the control group, 10D5 group, IgG2a group, and PD98059 group were 6.6% ±1.4%, 30.6% ± 2.4%, 8.1% ± 1.3%, 36.0% ±4.0%, respectively, with a significant difference among the four groups (F = 105.4, P ＜0.05 ). There was a significant difference in the expression of caspase-3 and Bcl-2 among the four groups (F=292.1, 181.6, P＜0.05). Conclusion Integrin αvβ6 can mediate the tolerance of gastric cancer AGS cells to 5-fluorouracil through direct β6-ERK binding.