Objective　To study the effects of MIP-1β and TGF-β antibody on CD34+ cells from cord blood in stroma-contact culture system. Methods　Immunomagnetic selected CD34+ cells were inoculated onto the pre-established irradiated human stroma layer. MIP-1β, TGF-β antibody (20 μg/ml) and MIP-1β+anti-TGF-β (5 μg/ml) were added on day 0 and day 5. On day 5 and day 10, cell counting, hematopoietic progenitor cells count were made by semi-solid culture and CD34+ cells were assayed by FACS. Results　On day 5, no significant difference of total cell number was observed as compared with the control group (P>0.05), but the number of CD34+cells, CFC, and HPP-CFC in TGF-β antibody and MIP-1β+ TGF-β antibody groups was significantly higher than that in the control (P<0.05). On day 10, the number of total cell, CD34+ cells, CFC, and HPP-CFC in TGF-β antibody and MIP-1β+ TGF-β antibody groups was significantly higher than that in the control (P<0.05). No significant difference was observed between groups MIP-1β and control either on day 5 or day 10 (P>0.05). Conclusion　MIP-1β has no significant effect on the CD34+ cells as compared with the control while the CD34+ cells can be expanded 1-3 folds with TGF-β antibody (20 μg/ml) or MIP-1β+ TGF-β antibody (5 μg/ml) in 10 days. There are synergic interactions between MIP-1β and TGF-β antibody.

Purpose　The aim is to investigate the effects of BSP on immunological function in normal and immunosuppressed mices.Methods　Thymus and spleen indexes, peripheral blood WBC number,the lymphocyte proliferation response, IL-2 production and serum and splenocyte hemolysin contents were measured after intraperitoneal injection of BSP in normal and immunosuppressed mices.Results　(1)BSP 100 mg/（kg*d）×10d significantly increased the thymus and spleen indexes and peripheral blood WBC number in immunosuppressed mice.The thymus and spleen indexes in normal mice was also increased. In addition,BSP markedly improved T,B lymphocyte proliferation responses and IL-2 production in normal and immunosupressed mices.(2) BSP improved the serum and splenocyte hemolysin contents in normal and immunosuppressed mice. Conclusion　It was suggested that BSP was a kind of immunomodulator, and could improve the immunological function of normal and immunosuppressed mices.

Objective To investigate the effects of adrenocorticotropic hormone (ACTH)combined with Huaiqihuang on hypothalamic pituitary adrenal (HPA) in rats. Methods Fifty rats were randomly divided into five groups according to the random number table method:normal control group (group A), prednisone group (group B), Huaiqihuang group (group C), ACTH group (group D) and combined treatment group (group E) with 10 rats in each group. Rats in group B, C, D and E were gavaged by acetic acid prednisone water solution 12.5 mg/(kg · d) for 4 weeks to establish HPA axis suppression model. Group A was given distilled water 10 mL/(kg·d) as control. Rats in group C and E were gavaged with Huaiqihuang 5 g/(kg·d) 30 minutes after intragastric administration of prednisone acetate. At the third week of the experiment, group D and E were subcutaneous injected with ACTH 200 μg/(kg·d). The serum cortisol levels were measured respectively at the start of the ex?periment, 2 weeks and 4 weeks of experiment. Animals were sacrificed at the end of the experiment, and then weights of the pituitary, adrenal glands and the viscera index were calculated. The pathological changes of the pituitary and adrenal glands were observed by HE stainning. Results After 2 weeks, the serum cortisol levels were significantly lower in group B, C, D and E than those of group A (P<0.05), suggesting that the model was successful. After 4 weeks, the serum cortisol levels were significantly higher in group C, D and E than those of group B (P<0.05), and between the treatment group the value was group E>group D>group C (P<0.05). At the same time, the weights of pituitary and adrenal gland and the viscera in?dex were higher in the three groups than those of B group (P<0.05). The HE staining showed that there were no significant changes in the distal part of the pituitary gland in five groups. The adrenal cortex zona was thinning and the structure was dis?ordered in group B. There were different degrees of hyperplasia in group C, group D, and group E, which was the most obvi?ous in group E. Conclusion ACTH combined with Huaiqihuang can promote adrenal cortex zona hyperplasia and cortisol secretion, which reduces the glucocorticoid induced inhibition of HPA axis in rats.

Objective To investigate the expression and function of transforming growth factor (TGF)-β1 and Smad2 in liver fibrosis of biliary atresia (BA). Methods Liver biopsy specimens were collected from autopsy (normal group, n=5), congenital biliary dilatation (CBD group, n=10), BA patients underwent Kasai procedure (early hepatic fibrosis group, n=19) and liver transplantation (transplantation group, n=11). The first three groups were collected from January 2010 to July 2014 in Tianjin Children’s Hospital, and the last group was collected from January 2013 to January 2014 in Tianjin First Central Hospital. The hematoxylin and eosin (HE) stain were used to observe the degree of liver fibrosis of four groups. Immunohistochemistry (IHC) was used to observe expressions of TGF-β1 and Smad2 in liver tissues of these samples. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to test the quantitative mRNA of TGF-β1 and Smad2 in these samples. Results Results of HE showed that no fibrosis in autopsy group, mild fiber cell hyperplasia in CBD group, severe fibrosis in Kasai group and significant pseudolobule in transplantation group. Results of IHC showed that TGF-β1 was expressed in the cytoplasm of hepatocytes, bile duct cells, lymphocytes and neutrophils. The average optical density of TGF-β1 was the highest in Kasai group compared with that of other three groups (P < 0.05). There was no significant difference in Smad2 expression in cytoplasm of hepatocytes, bile duct cells and lymphocytes between four groups (P>0.05). Results of qRT-PCR showed that both TGF-β1 mRNA and Smad2 mRNA were the highest in early hepatic fibrosis group than those of CBD group and transplantation group (P<0.017). Conclusion In early stage of BA, TGF-β1 and Smad2 promote liver fibrosis until the formation of P-P,P-C desmosome structure. However, with BA fibrosis becomes more serious, the pro-fibrogenic function of TGF-β1 and Smad2 becomes less.

Objective To observe the counteraction of Ganoderma lucidum polysaccharide(GLP) on the inhibition of mice splenocyte proliferation and IL-1? and IL-2 mRNA expression induced by prostaglandin E2(PGE2).Methods Mixed lymphocyte culture reaction(MLR) method was used for the experiment.The lymphocyte proliferation was determined by MTT method,and the levels of IL-1? and IL-2 mRNA expression were evaluated by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results After co-culture with PGE2 for 48 hours,the splenocyte proliferation was inhibited to some extent,and the difference was significant when the concentration of PGE2 was over 10?mol/L(P

Objective: To investigate the effects of BSP on lymphocyte function and cytokines production in normal and immunosuppressed mice.Methods: The lymphocyte proliferation response, T lymphocyte subpopulations and IL 2 production were measured after intraperitoneal injection of BSP in normal and immunosuppressed mice, and the activity of IL 1 and TNF secreted by mouse peritoneal macrophages were detected after BSP supplementation in vitro.Results: (1)BSP〔(100 mg/(kg?d)?10 d)〕markedly improved T,B lymphocyte proliferation responses and IL 2 production in normal and immunosuppressed mice, and significantly increased the number of L 3T + 4 cells in thymus and spleen of immunosuppressed mice, and the ratio of L 3T + 4/Lyt 2 + cells was also increased.(2)BSP distinctly enhanced the activity of IL 1 and TNF production by mouse peritoneal macrophages in vitro.Conclusion: BSP can improve the immunological function of normal and immunosuppressed mice.

Objective To investigate the effect of As2 O3 in combination with cinobufacini on proliferation and apoptosis of the K562 cells and provide theoretical basis for clinical application.Methods Cell proliferation was assayed by analyzing the growth and viability of the cells.Apoptosis was assayed by performing cell morphology,Annexin-V/PI staining,DNA-PI staining,and DNA gel electrophoresis.Results After exposure to As2O3 and cinobufacini,the growth of K562 cells was inhibited and the viability of K562 cells was decreased. After treated with 1.0μmol/L As2O3,0.125μg/ml cinobufacini,0.25μg/ml cinobufacini,1.0μmol/L As2O3 + 0.125 μg/ml cinobufacini,1.0μmol/L As2O3 + 0.25μg/ml cinobufacini for 24 and 48 hours,the proliferation inhibition rate were(24 ± 1.3)%,(21 ± 1.5)%,(38 ± 3.1)%,(57 ±2.7)%,(66 ±3.3)% and(49 ±2.9)%,(48 ±2.7)%,(61 ±2.1)%,(77 ±3.8)%,(82 ±4.2)%,the apoptosis rate of K562 cells were(4.8 ± 0.5)%,(5.6 ± 0.7)%,(9.8 ± 0.6)%,(11.9 ± 1.2)%,(15.2±1.5)% and(11.0 ±0.9)%,(12.9 ±1.1)%,(18.4 ±1.5)%,(21.0 ±2.0)%,(28.0 ±1.9)%.The percentage of apoptotic cells was a time- and dose-dependent manner.Typical DNA ladder was shown by DNA gel electrophoresis.Conclusions As2O3 combined with cinobufacini inhibited the proliferation of K562 cells and induced apoptosis of the K562 cells,the combination of the two drugs had better effect.

Objective To study the imaging appearances of congenital splenorenal venous shunt,and to evaluate their clinical significances.Methods Of 127 283 patients who underwent upper abdominal CT scanning,6 patients were diagnosed to have congenital splenorenal venous shunt.The imagings were studied retrospectively.Plain scanning,enhanced CT and Doppler ultrasonography were performed on all these patients.Results The incidence was about 47/million.The six patients were all females,the age was between 32 to 67 years.The mean age was 48.8 years.Enhanced CT demonstrated that there were twisted and dilated shunting blood vessels between the splenic and renal veins.The diameters in four patients were larger than the splenic and portal veins.In the remaining two patients,they were smaller shunting blood vessels.One patient had an associated absence of right portal vein.Two patients had associated dysplasia of portal veins and splenic veins.MPR,CPR,MIP and VR could three-dimensionally depict the courses,the beginnings and the ends of the splenorenal shunts.Doppler ultrasonography showed counterflow between the portal and splenic veins,and showed the blood in the splenic vein to flow into the left renal vein.Conclusions A congenital splenorenal venous shunt is one of the rare form of congenital extrahepatic portosystemic venous shunts.Enhanced MSCT scan combining with its post-processing techniques could clearly demonstrate the shunt vessels and the associated lesions.Doppler ultrasonography could further demonstrate the shunt direction.