Objective: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. Methods: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-? and TNF-? in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. Results: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-? and NO in Gl-SP-or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-? in double-way MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4+ or CD8+ subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4+ and CD8+ subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. Conclusion:Gl-SP and Gl-BSP have sim-ilar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.

Objective　To investigate the role of cytotoxic T lymphocyte associated molecule-4Ig(CTLA-4Ig) in the prevention of C57BL/6 mice autoimmune hepatitis. Methods　The C57BL/6 mice were intraperitoneally immunized with C57BL/6 mice liver-specific protein in complete Freund's adjuvant. At the same time CTLA-4Ig were given to observe the pathologic alteration of C57BL/6 mice liver. Results　With the increase of time of immunization, the results in the treatment group were similar to those of the control group; but inflammatory cell infiltration, hepatic cell swelling, focal necrosis and severe hepatocyte damage were found in the pathologic model group. There was a significant difference between the pathologic model group and control one. Conclusion　Autoimmune hepatitis of C57BL/6 mice can be effectively prevented by CTLA-4Ig.

Objective To investigate the effects of soybean peptide calcium on apparent calcium absorption rate and bone mineral density of rats.Methods Totally 50 male Wistar rats were divided into 5 groups at comparable body weight:the low calcium control group,calcium carbonate control group and 3 groups with low,medium and high dose of soybean peptide calcium(SPC).Rats of each group were fed with designed forage.At the end of the fourth week of the feeding.calcium metabolic experiment was performed for 3 days and the apparent calcium absorption rate was calculated.Three months later,the left and right femurs of the rats were stripped to assay the bone calcium and bone density.Results Apparent calcium absorption rate of the calcium carbonate control group was 69.9 ％,which was lower than medium(82.6 ％) and high doses(77.4％) of soybean peptide calcium groups(F =31.371,P=0.000,0.001).Compared with the low calcium control group,rats with medium and high doses of SPC had obviously increased levels of weights(F =3.150,P =0.010,0.002),bone mass(F =3.195,P =0.015,0.002),calcium content (F=4.506,P =0.004,0.001)and bone mineral density of midpoint (F=6.185,P =0.006,0.000) and far end of femur (F =6.081,P=0.027,0.000).Despite of the weight index (P＞0.05),the high dose of soybean peptide calcium group also had the same results as the calcium carbonate control group (P=0.046,0.024,0.013,0.001).Conclusions Soybean peptide calcium can promote the apparent calcium absorption rate and increase the bone mineral density.

Objective To investigate the influencing conditions of human epidermal stem cells(ESCs) differentiating into gland-like cells and to identify the induced tubular structure.Methods The ESCs were seeded onto compound polysaccharide shell dermal matrix and collagen gel,adding different concentrations of epidermal growth factor(EGF) and culturing by vertical shaking in vitro,the three dimensional culture and induced directional differentiation were performed.The means of HE staining,immunofluorescence and confocal laser scanning microscope were adopted to observe the conditions,morphology and phenotype change of ESCs directionally differentiating into glandular epithelium-like cells.Results 15-20 ng/mL EGF could induce ESCs in tissue-engineering dermis to grow into dermis and appear the gland-like structure.The HE staining in this structure showed its profile as a single layer with lacune in the middle compartment,eosinophilic cytoplasm,lightly stained.Under CLSM,by CK19 staining,the luminal structure in the middle of the cell mass was observed,while CK18 and CEA were expressed in this structure.Conclusion Under the induction of particular concentration of EGF,the in vitro cultured human ESCs seeded on the tissues engineering dermis could form into the tubular structure,which is similar to the in vivo sweat gland secretory cells in morphology and histology.

Object To study the amount and properties of glycopeptide of Ganoderma lucidum (Leyss ex Fr ) Kaist. cultivated with grass (GLPG) and wood log (GLPW) ; to investigate the feasibility of substitution of grass for wood log to cultivate G lucidum Methods Glycopeptides were extracted and isolated from the fruit body of G lucidum followed by dialysis; the monosaccharid constituent and relative molecular weight of polysaccharides were examined by GC and gel filtration; structure of glycopeptide was determined by IR, 1H NMR and 13 C NMR Results Relative molecular weights of GLPW and GLPG glycopeptides were (Mw) 5 13?10 5 and 5 85?10 5, (M?) 6 21?10 5 and 6 96?10 5, respectively; the glucosidic bonds of both glycopeptides were ? type; both glycopeptides consist 16 amino acids and Rha, Xyl, Fru, Gal and Glc (there was trace amount of Man in GLPW) Conclusion There are less physical and chemical difference between GLPG and GLPW The purification rate of GLPG is 2 8 times to GLPW

Objective:To find out the effects of Ganoderma lucidum polysaccharides peptide (Gl-PP) on the invasion of the human lung carcinoma cell (PG cell). Methods: PG cells were pretreated with different concentration Gl-PP in vitro, using cell proliferation assay, cell migration assay, adhesion assay, zymography and RT-PCR, then the effects of Gl-PP on proliferation, motility, adhesion and MMP-9 activity and mRNA expression of PG cells were investigated in vitro. Results: Gl-PP did not directly inhibit PG cell proliferation in vitro. However, pretreated with Gl-PP, PG cells motility was inhibited significantly. PG cells adhesion was also inhibited. The activity of MMP-9 was inhibited in a dose-dependent manner, and the inhibited ratio was 41.53% at a dose of 100 mg/L Gl-PP. The mRNA expression of MMP-9 of pretreated PG cells was inhibited. Conclusion: Gl-PP could suppress invasion of human lung carcinoma cells in vitro.

Objective To report the diagnosis and treatment of the pregnant women, who were positive for syphilis in serological test, as well as their final pregnant results. Methods we conducted a retrospective analysis of the serological test results of 3 105 pregnant women subjected to childbirth or induced labor in the department of gynaecology and obstetrics at our hospital since January of 2004. Results Of the 3105 pregnant women, 21 (6.76 ) were positive for treponema pallidum (TP). Additionally, 7 (33.33%) cases were positive for TRUST, and 6 (28.57%) negative. Only 2 cases showed skin lesion, canker and soft chancre. Of the 18 cases, who were found to get infection during pregnancy, 11 (61.11%) had taken the treatment against syphilis. Of the 21 women positive for TP, 5 were subjected to induced labor, 13 to cesarean section, and 3 to spontaneous parturition. The average time of pregnancy was (267.44?11.00) d. The Apgar score of all 16 newborns was 10 at birth. The umbilical blood of 5 infants was tested by TP and TRUST, among which 3 were positive by TP, 2 were positive for TRUST. All infants did not have any clinical manifestation of congenital syphilis after birth. There were 5 cases transferred to the ward of pediatrics, among which 2 undertook the treatment against syphilis, 2 against premature delivery, and 1 against asphyxia, and finally all had been cured. Conclusion The patients, who were found positive for TP during the early and middle period of pregnancy, can obtain a fine pregnant result, if undertaking regular examination and treatment against syphilis.

Objective To compare the monosaccharide composition and molar ratio of GL-PPT2, GL-PPT3, and GL-PPT4, which are three polysaccharide peptides isolated from the fruit body of Ganoderma lucidum. Methods The polysaccharide peptides were hydrolyzed by triflouroacetic acid, then derivatized by trichloro-aldehyde-1-pheny-3-methyl-5-pyrazolone (PMP), and determined by HPLC method with the 245 nm detection wavelength. Gradient elution with the solution of KH2PO4 (pH 5.0)-acetonitrile (83.5∶16.5) resulted in baseline separation of nine monosaccharides, of which the chromatographic peaks were very clear. Results GL-PPT2, GL-PPT3, and GL-PPT4 comprise the nine monosaccharides of mannose, ribose, rhamnose, glucose, galactose, arabinose, xylose, galactose, and glucuronic acid. Conclusion The ribose in polysaccharide peptides of G. lucidum is first reported.

Objective To evaluate the clinical significance of bladder saline perfusion before catheter removal in TURP patients. Methods From 2009 to 2011,140 patients received TURP were enrolled in this study.Patients were divided into perfusion group (70 cases with bladder saline perfusion before catheter removal) and control group (70 cases without perfusion). Results Comparing with the control group (33.1 ± 5.4) min,the time waiting for urination was shorter in perfusion group ( 3.7 ± 0.2 ) min ( P ＜0.05 ).The recovering time to normal urination was shorter in perfusion group (7.7 ± 1.2 ) d than in control group (11.7 ± 1.3) d (P ＜ 0.05 ) as well.In the first urine after catheter removal and first urine on the next day morning,white blood cell count of 2 groups (4.5 ± 0.1 ) vs ( 6.9 ± 3.5 ) ; ( 3.7 ± 0.2 ) vs (4.3 ±0.5) had significant differences ( P ＜ 0.05 ). Conclusion Bladder saline perfusion before catheter removal in TURP patients is simple and effective for the restoration of normal voiding.

Background:The overlap of autoimmune liver disease with extrahepatic autoimmune disease adds the difficulty of diagnosis. It is essential to understand their clinical features. Aims:To analyze the clinical characteristics of primary biliary cirrhosis-autoimmune hepatitis( PBC-AIH ) overlap syndrome, and to investigate the effect of extrahepatic autoimmune disease on PBC-AIH. Methods:The clinical data including general information,clinical manifestations, biochemical parameters,immunological parameters,liver histopathological features,imaging findings and incidence of extrahepatic autoimmune disease of 81 patients with PBC-AIH were retrospectively analyzed. According to whether overlapping with extrahepatic autoimmune disease,PBC-AIH patients were divided into combination group and non-combination group. The clinical characteristics of the two groups were analyzed. Results:The ratio of male to female of 81 patients with PBC-AIH was 1 ∶ 9. 1,and the average age was(58. 1 ± 9. 9)years old. Of the patients 53. 1% had symptoms,the percentage of jaundice and pruritus was 18. 5% . Serum levels of ALT,AST,ALP,GGT,TBil and Glo were increased in all patients and IgG and IgM levels were increased in most of the patients. The positive rates of ANA, AMA and/ or AMA-M2,SMA and gp210 and/ or sp100 were 100% ,90. 1% ,1. 2% and 11. 1% ,respectively. Portal lymphoplasmacyte infiltration and interface hepatitis were the major liver histopathological features;50. 6% of the patients had both histological features of PBC and AIH. Cirrhosis was diagnosed in 28 patients(34. 6% )by imaging,and abdominal lymph nodes enlargement was found in 76 patients(93. 8% ). Twenty-three patients(28. 4% )overlapped with extrahepatic autoimmune disease;Sjogren’s syndrome and autoimmune thyroid disease were the most frequently overlapped extrahepatic autoimmune diseases. The level of GGT was significantly lower in combination group than in non-combination group(P = 0. 001). No statistically significant differences in other clinical characteristics were found between the two groups. Conclusions:It is crucial to understand the clinical characteristics of PBC-AIH and to screen the overlapped extrahepatic autoimmune disease,especially Sjogren’s syndrome and autoimmune thyroid disease.