ObjectiveThe aim of this study is to investigate whether oral administration of collagen Ⅱ peptide (250-270)[C Ⅱ (250-270)]-cholera toxin B subunit (CTB)complex could effectively set up oral immune tolerance to collagen-induced arthritis (CIA) in mice. MethodsDBA/1 mice were divided into Ⅰ, Ⅱ, Ⅲ and Ⅳgroups. Group Ⅰ was normal control group. Collagen type Ⅱ emulsified in Freund's complete adjuvant were injected to mice of groups Ⅱ , Ⅲ and Ⅳ twice from the base of the tail. Mice of group Ⅲ were fed with C Ⅱ (250-270)-CTB covalent complex twice after the arthritis was developed. Mice of group Ⅳ were fed with C Ⅱ(250-270) and CTB mix at the 14th day after primary immunization. Visual scores and histopathologic scores of arthritis were recorded. The frequencies of arthritis between the groups were compared usingFisher's exact test. The clinical and histological severity of arthritis were analyzed by ANOVA.Results The frequencies of arthritis in groups Ⅰ , Ⅱ , Ⅲ and Ⅳ were 0, 100％, 100％ and 25％ respectively. Average accumulative scores of arthritis were 0, 5.0±1.7, 10.8±2.8 and 1.0±2.0 respectively. Average accum-ulative histopathological scores of arthritis were 0, 16±8, 32±13 and 7±6 respectively. Conclusion Oral administration of C Ⅱ (250-270) and CTB mix in arthritis mice after C Ⅱ immunization can suppress the onset and severity of arthritis. Oral administration of C Ⅱ (250-270)-CTB covalent complex in the acute stage of arthritis can accelerate arthritis.

BACKGROUND:Sensorineural hearing loss is mainly caused by missing or damaged hair cells in the inner ear. Application of adipose-derived mesenchymal stem cells to regenerate inner ear hair cells is an effective treatment for hearing loss. OBJECTIVE:To explore the feasibility of in vitro inducing adipose-derived mesenchymal stem cells to differentiate into inner ear hair cel-like cells in guinea pigs. METHODS:Adipose-derived mesenchymal stem cells from guinea pigs were isolated and cultured to the 3rd generation. cellphenotype was detected using flow cytometry. Cytokines were added for induction and differentiation by stages, including epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor-1, al-trans retinoic acid, brain-derived neurotrophic factor, neurotrophin 3. RESULTS AND CONCLUSION:Adipose-derived mesenchymal stem cells of guinea pigs cultured in vitro were fusiform and showed a swirled adherent growth. Passage 3 cells were positive for CDCD29 and CD44, but negative for CD34 and CD45. After induction, the cells were positive for nestin and GFAP positive at early stage;after 10-day continuous induction, the cells expressed Myosin VIIa and Math1, specific markers of hair cells, indicating that cytokines can directly induce adipose-derived mesenchymal stem cells to differentiating into inner ear hair cells in guinea pigs.

Objective To optimize the extraction process and to establish a method for the content determination of the polysaccharides in Ophiopogonjaponicus.Methods With extracts rate of polysaccharide as the index,an orthogonal de- sign test was adopted to optimize the extraction process.And phenol-sulphuric acid colorimetric method was used to de- termine the content of polysaccharides in Ophiopogonjaponicus.Results The optimum extract conditions were as follows: refluxing twice with eight times amount of water,thirty minutes each time,and precipitation with 80 % alcohol.A good linearity of polysaccharides was in range of 0.02032～0.10008 mg(r=0.9998),and the average recovery rate was 102.41%,RSD was 1.78 %.Conclusion The polysaccharide can be fully extracted under these conditions.

Objective To observe the changes of intedeukin (IL)-17 in patients with maintenance hemodialysis (MHD) and its relationship with anemia. Methods Twenty sera from normal people(normal group) and sixty-eight sera from patients with MHD were sampled. Patients with MHD were divided into 4 groups by haemoglobin (Hb) levels: severity anemia group (12 cases), midrange anemia group(18 cases),light anemia group(18 cases) and non-anemia group(20 cases). IL-6 and IL-17 levels were measured in all the sera with ELISA. Meantime, C-reactive protein (CRP),Hb, ferritin and iron saturation ratio was observed. Results CRP,IL-17 and IL-6 levels in MHD patients were much higher than those in normal group. IL-17 levels in severity and midrange anemia group were higher than those in light anemia group,and IL-17 was positively correlated to Hb. Conclusions IL-17,IL-6 and CRP in patients is higher than that in normal people,and the microinflammation status exists in MHD. IL-17 may play a role in pathogenesis of microinflammation in MHD patients with anemia.

The clinical data and routine biochemical parameters of 64 maintenance hemodialysis (MHD)patients and 20 controls were analyzed in the study.Serum cystatin C levels were detected by latex particle enhalice inununo-turbidimetry:and the cardial structure and function were assessed by echocardiography.As MHD time extended,the levels of semm cystatin C increased gradually,accompanied with high incidence of left ventricular hypertrophy(LVH).The LVH-positive patients had higher systolic blood pressure and left veHtricular mass index,and had higher serum cystatin C than tllose in LVH-negative patients.The serum cystatin C levels were positively correlated with left yentricular mass index(r=0.633,P<0.01)and systolic blood pressure(r:0.397,P<0.01).The results suggest that serum cystatin levels may be an influencing factor for long-term cardiacvascular complication in MHD patients.

Objective To observe the effects of mesenchymal stem cells (MSCs) surface CXC chemokine receptor 4 over-expression on the repair of kidney after ischemia reperfusion (I/R) injury.Methods The MSCs were co-cultured with I/R injured renal cell homogenate supernatant.The MSCs surface CXCR4 and stromal cells derived factor-1 (SDF-1α) protein levels were detected by Western blot, chemotactic ability of MSCs to SDF-1 was investigated by transwell test.The I/R injured renal model was made and pathological changes were observed in control group, I/R group, MSCs injection group and CXCR4 neutralize antibody group.Renal CXCR4 protein expression was measured by immunofluorescence histochemistry, SDF-1α、 CXCR4、 hepatocyte growth factor (HGF) and epidermal growth factor (EGF) mRNA were detected by Real-time quantitative polymerase chain reaction (RT-PCR).Comparisons among multiple groups were performed using One-way analysis of variance, and comparisons between groups were carried out using independent-sample t-test.Results In vitro, the SDF-1α protein expression markedly increased in I/R injured renal tissue homogenate, but the difference was not significant between I/R group and CXCR4 antibody group (t =0.862, P =0.403).MSCs surface CXCR4 protein expression increased significantly after co-cultured with I/R injured renal tissue homogenate (F =95.957, t =10.166, P ＜ 0.01), and the chemotactic ability of MSCs to SDF-1 increased at the same time (F =82.459, t =6.826, P ＜ 0.01), the CXCR4 protein expression (t =13.657, P ＜ 0.01) and the chemotactic ability (t =12.662, P ＜0.01) could be decreased by CXCR4 neutralize antibody.In vivo, renal tubular structure was destroyed in I/R group.After MSCs injection, the renal pathological injury improved rapidly, but the improvement could be inhibited by CXCR4 antibody.The expression of SDF-1α mRNA and level of SDF-1a protein increased in I/R group, but there was no significant difference among different groups (F =1.909,P =0.173).MSCs injection markedly up-regulated the CXCR4 protein and mRNA expression (F =6.663, P =0.006).Following the increase in CXCR4 expression, the expressions of HGF mRNA (F =11.898,P ＜ 0.01) and EGF mRNA (F =5.309, P ＜ 0.05) increased gradually which could be restrained by CXCR4 antibody (t =5.312, t =4.310, P ＜ 0.01).Conclusions I/R injured renal microenvironment markedly increased the mesenchymal stem cells surface CXCR4 expression, and increased CXCR4 expression can induce MSCs chemotaxis and stimulate the secretion of renal protective growth factors paracrine promoting the repair of the kidney.

BACKGROUND:Can adipose-derived mesenchymal stem cells be used to treat sensorineural deafness caused by hair celldegeneration and loss? OBJECTIVE:To investigate the effect of adipose-derived mesenchymal stem cells from guinea pigs transplanted via the scala tympani on the recovery from sensorineural hearing in an animal model. METHODS:Intraperitoneal injection of gentamicin was performed to prepare sensorineural deafness models in guinea pigs. Then, adipose-derived mesenchymal stem cells from guinea pig were transplanted via the scala tympani. After 1 and 3 weeks, auditory brainstem responses were detected to observe the hearing variation after adipose-derived mesenchymal stem celltransplantation. Meanwhile, the migration and distribution of EDU-labeled adipose-derived mesenchymal stem cells in the cochlea were traced. RESULTS AND CONCLUSION:After 1 and 3 weeks of implantation, the results from auditory brainstem response showed that the hearing was gradual y improved. At 1 week of implantation, most of the cells were distributed in the perilymph and some cells migrated to the Corti organ and attached to the basement membrane. At 3 weeks of implantation, cells migrated and attached to the basement membrane of Corti organ, furthermore, some cells migrated to the cochlear nerve. The longer the time of implantation, the less cellsurvived. The results indicate that adipose-derived mesenchymal stem cells from guinea pig that are transplanted via the scala tympani can directly migrate and survive ultimately to improve the hearing.

Objective To investigate the correlation between methylation status and death-associated protein kinase 1 (DAPK1) in SiHa and Hela cell line of cervical carcinoma and intervention of DNA methyltransferase inhibitor 5-azacytidine on the expression of DAPK1 and the proliferation of the cells. Methods DAPK1 methylation status was analyzed using methylation-specific PCR methods. The expression of mRNA and protein of DAPK1 were analyzed by RT-PCR and SABC methods after the treatment with 5-azacytidine. MTT assay was used to observe the changes of proliferation activity of the cells after 5-azacytidine treatment. Results DAPK1 genes were methylated and did not express in SiHa cells in the cervical carcinoma. Its expression could be restored by 5-azacytidine. MTT assay showed 5-azacytidine could weaken the proliferation of cancerous cells. Conclusion DAPK1 methylation plays an important role in the carcinogenesis of cervical cells and can reexpress after the treatment with 5-azacytidine which also restored its inhibitory function on carcinoma.

OBJECTIVE:To Evaluate the characteristics of national essential drugs on the view of pharmaceutics.METHODS:Referring to related literatures,the characteristics of national essential medicines were analyzed and summarized on the basis of pharmaceutics.The national essential drugs list was compared with that of WHO's.RESULTS:In the equipment part of primary healthy care institution national essential drugs list (2009 edition),chemical and biological medicines had 205 species which included 270 formulations,were classified as 21 kinds of dosage forms.Chinese traditional medicines had 102 species which included 188 formulations,were classified as 13 kinds of dosage forms.The ration of the amount of dosage forms to species is 1.49,a little higher than WHO(1.31) now.CONCLUSION:The dosage forms of national essential drugs were abundant and enable to meet the clinical requirement.But should optimize the breed and add in many species of dosage form to satisfy the needs of medicines useage from urban to rural.

To compare the changes of serum osteocalcin (OC), interleukin-18 (IL-18) in patients with different glucose tolerance and to explore their relationship to carotid atherosclerosis in type 2 diabetes mellitus (T2DM). Methods: According to the result of oral glucose tolerance test (OGTT), 140 research subjects were divided into 3 groups: Normal control group, n=50, Pre-diabetes group, n=30 and T2DM group, n=60 which included in 2 subgroups:Normal carotid intima-media thickness (IMT) subgroup, n=26 and Carotid IMT thickening subgroup, n=34. Carotid IMT, serum OC, IL-18 and glycosylated hemoglobin (HbA1c), fasting glucose, OGTT 2-hour blood glucose (2hPG), TC, TG, LDL-C, HDL-C, body mass index (BMI), insulin resistance index (HOMA-IR) and insulin cell function index (HOMA-β) were measured in all subjects. Pearson correlation and multi liner regression model were conducted to analyze the relevant parameters. Results: From Normal control to Pre-diabetes to T2DM groups, serum OC was decreased and IL-18 was increased gradually, all P<0.05. OC was negatively related to HbA1c (r=-0.426), fasting glucose (r=-0.582), 2hPG (r=-0.489), HOMA-IR (r=-0.456), TC (r=-0.451) and carotid IMT (r=-0.559), all P<0.05; while positively related to HOMA-β (r=0.439), P<0.05. IL-18 was positively related to BMI (r=0.395), HbA1c (r=0.693), fasting glucose (r=0.880), 2hPG (r=0.715), HOMA-IR (r=0.667), TC (r=0.734), TG (r=0.326), LDL-C (r=0.471) and carotid IMT (r=0.857), all P<0.05; while negatively related to HOMA-β (r=-0.678), P<0.05. In T2DM group, carotid IMT was positively related to IL-18 (r=0.817), fasting glucose (r=0.415), HOMA-IR (r=0.356), TC (r=0.396) and TG (r=0.362), all P<0.05; while negatively related to OC (r=-0.588), P<0.05. Multi liner regression analysis indicated that IL-18, OC, TC and fasting glucose were the independent impact factors for carotid IMT (regression coefficients were 0.013, -0.011, 0.044 and 0.044 respectively), P<0.05. Conclusion: Serum OC and IL-18 had been involved in glucolipid metabolism and closely related to the occurrence and development of carotid atherosclerosis in T2DM patients.