Ebosin is a novel exopolysaccharide produced by Streptomyces sp.139 with remarkable activity against rheumatic arthritis in vivo. In this paper, we reported effects of Ebosin on the inflammatory cytokines including interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNFalpha) in THP-1 cells. With the special fluorogenic peptide as substrates, the enzymatic activities of interleukin-1beta converting enzyme (ICE) and TNFalpha-converting enzyme (TACE) were inhibited by Ebosin separately. Using the real-time reverse transcription polymerase chain reaction (real-time PCR), the mRNA synthesis of the three cytokines were identified decline separately by Ebosin. The secretion quantum of three cytokines in THP-1 cells with Ebosin was lower than that of normal THP-1 cells determined by ELISA assay and Western blotting. All of these results showed that Ebosin has remarkably suppressed synthesis of the three cytokines in THP-1 cells through different pathways. The primary study of Ebosin on anti-inflammation mechanism was promoted developing the new drugs treating rheumatic arthritis.

0.05).Ambulation scores revealed significant difference (P

Objective To isolate,culture and identify epidermal stem cells(ESCs).The gene of enhanced green fluorescent protein(eGFP) was introduced via recombinant adeno-associated virus(rAAV) infection into the epidermal stem cells in vitro and the transfection efficiency under various tite of culture was determined. Methods 1.Getting fetal Wistar rats'dissociated single epidermal cells.2.Making laminin and CollagenⅣ the substitution of basal memrane,ESCs were isolated by adhering to murine laminin and CollagenⅣ.3.Epidermal stem cells were cultured in twenty-four-well plate,and cells number was controled 5?10~4 or so.After epidermal stem cells adhered to plate,diluted rAAV_2/eGFP virus fluid with serum-free medium.The diluted rAAV_2/eGFP virus fluid was added to the twenty-four-well plate according to the different MOI(viru gene/cells).The number of green fluorescence cells were counted under fluorescence microscope.The transfection efficiency was determined. Results 1.ESCs had better adhesive ability to laminin and higher colony formation efficiency(CFE) than that of keratinocytes.2.ESCs wer strongly positive with immunocytochemical staining of integrin ?1 and keration 19(K19).3.After rAAV_2/eGFP genetic transfection of ESCs,the positive cloning expressed highly eGFP gene along with time.Conclusion 1.These results suggest that rats'epidermal stem cells could been successfully isolated and enriched in vitro by means of rapid adherence to laminin and CollagenⅣ.After rAAV_2/eGFP genetic transfection of epidermal stem cells,the positive cloning expressed highly eGFP gene stably along with time.

Objective We compare the effects of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation in order to explore the relationship between three-dimensional scaffold and cartilage tissue engineering seed cells BMSCs differentiation. Methods BMSCs together with chitin and fibrin gel complexes were cultured in vitro and implanted into rat's articular cartilage defect location. After 14 days in vitro culture, HE staining, toluidine blue staining and type II collagen immunohistochemical staining were performed;After transplantation in vivo, for 2 weeks, 4 weeks and 6 weeks, morphology observations, expression of cartilage-specific protein analysis and BMSCs in vivo tracer method were performed. We analyzed the differentiation of BMSCs into chondrocytes by statistical methods. Results The comparison of positive cell rate of type II collagen immunohistochemical staining in BMSCs-fibrin gel group and BMSCs-chitin group which were cultured in vitro, showed no significant difference with control group. Integrated absorbance(IA) change rate of toluidine blue staining and type II collagen immunohistochemical staining in BMSCs-fibrin gel group which was cultured in vivo, showed significantly different with other groups and control group.Conclusion The results showed that in vitro fibrin gel or chitin has very weak induction of BMSCs to cartilage differentiation, while in vivo BMSCs-fibrin gel can facilitrate BMSCs to differentiate into chondrocyte-like cells.

Data were analyzed from self-filled questionnaires for the students who took the elective course of general practice in China Medical University. 95.5% of students thought it was necessary to study general practice in the college. 0-10 was used to score the degree of how they want to choose general practice as a career, the median was 0 before they had the class and the median was 5 after the class (Wilcoxon rank test, P<0.01). But there were still some places need to be improved in terms of curriculum design, teaching methods and community practice. The General Practice teaching need to be innovated and the teaching mode should be different from the traditional one.

The high mobility group A2 (HMGA2),one of non-histone chromatin protein,can specificly bind to AT-rich DNA sequences by its AT-hook and work as oncofetal gene and architectural transcription factor.HMGA2 expresses in almost all kinds of malignant neoplasms,which is closely related to the formation,development and poor prognosis of the neoplasms.HMGA2 plays a very important role in every biological process including cell proliferation,cell cycle,stem cell self-renewal,epithelial-mesenchymal transition and DNA damage repair.It is of great significance to study the effect and mechanism of HMGA2 in neoplasms.

BACKGROUND: Tissue engineering method has been employed to recover cartilage defect and overcome many traditional shortages.OBJECTIVE: In vitro marrow mesenchymal stem cells (MSCs) of adult rat was induced to differentiate into chondrocyte phenotype so as to probe into the feasibility of MSCs to be cartilage seed cell in tissue engineering.DESIGN: Completely randomized design and controlled experimental study.SETTING: Department of Histology and Embryology, and Department of Neurobiology, Harbin Medical University MATERIALS: Six Wistar rats of either gender, cleanness grade, were provided by Experimental Animal Center, Affiliated Second Hospital, Harbin Medical University. Permission number of experimental animal production was SCXK(black) 20020002.METHODS: The MSCs of the second generation adult rat was taken and divided into test group and control group. The test group was induced with serum free and control group was induced with 10% fetal calf serum. Collagen type Ⅱ immunohistochemical staining, toluidine blue staining was performed to detect differentiation.MAIN OUTCOME MEASURES: Identification of chondrocyte, comparison of the positive rate of Collagen type Ⅱ immunohistochemical staining at different induction time point.RESULTS: Induced MSCs had identical characteristic to the chondrocyte.CONCLUSION: In vitro culture can induce MSCs to differentiate directly to chondrocyte-like cell. The results suggest that it is feasible to use MSCs as seed cells in the cartilage tissue engineering.

Objectives To summarize screening and therapeutic effects of congenital hypothyroidism (CH) in Zhongshan. Methods The thyroid stimulating hormone (TSH) concentration in dried heel blood samples on iflter paper was detected using time-resolved fluorescence immunoassay. The cases of positive screening tests were called back for further examination of venous blood TSH concentration using chemiluminescence method. Fifty-four children with permanent CH treated routinely for 2 years (CH group) and 120 age-gender matched health children (control group) were recruited. The physical development (height, body weight) was monitored. The neurodevelopment and temperament type were tested using Pediatric Nneuropsychological Development Assessment and Children's Temperament Scale respectively at 6 and 24 months after birth. Results Two hundred eight-five thousand two hundred forty-two neonates were screened. One hundred and forty cases were confirmed and the incidence rate was 1/2037. There was no statistical difference in length-for-age z score (LAZ) and weight-for-age z score (WAZ) between CH and control group (P>0.05). The neurodevelopment in CH group was normal, but gross motor development was worse than that in control group (P<0.05). The temperament type and distribution had statistical difference between CH and control group (P<0.05). The percentage of the dififcult type and central dififcult type was increased in CH group as compared to control group, especially in the activity, adaptability, reaction intensity and perseverance (P<0.05). Conclusions The physical and neurodevelopment are nearly normal in patients with CH after early supplementation, but the psychological behavior problems need to be focused on in the process of intervention.

Objective To investigate the clinical significance of testing serum kisspeptin in central precocious puberty (CPP) girls. Methods Sixty eight CPP girls and 68 healthy girls was studied from December 2012 to December 2014. HEK293 cells were cultured. Luciferase reporter assay was performed to verify the binding of miR-137 to the 3′UTR of KISS1. Serum miR-137 level was levaluated by qRT-PCR. Level of serum luteinizing hormone , prolactin , follicle stimulating hormone , thyrotropin , free thyroxine and estradiol was evaluated by chemi-luminescence immunoassay. The level of serum kisspeptin was detected by ELISA. Results MiR-137 was confirmed to bind to the 3′UTR of KISS1. The level of serum miR-137 was downregulated and kisspeptin was enhanced in CPP girls. The expression of miR-137 and kisspeptin was negatively correlated. Serum miR-137 level was negatively related to bone age and bone age advancement. According to the results of GnRH stimulating test, serum miR-137 was related to peak LH and peak/basal LH ratio. Conclusions MiR-137 could bind to the 3′UTR of KISS1. MiR-137 may be a potential biomarker for CPP assisted diagnosis.

With IL-6R as target, a new compound 2460A was identified from fungus using HTS screening model. The taxonomics of the produced strain was confirmed to be Trichoderma hazianum rifai after sequencing analysis of rDNA-ITS (internal transcribed spacer). Results showed that this compound has a binding activity on IL-6R competed with IL-6, thus it is a new ligand of IL-6R originating from microbe. With MTT assay, the anti-tumor activities of 2460A were demonstrated on CM126 and HT-29 cell lines separately, the IC50 are 2.17 x 10(-5) mol x L(-1) and 1.8 x 10(-5) mol x L(-1) respectively. The compound affected lightly the HT-29 cell cycle at S phase. Studies for the anti-tumor activity of 2460A in vivo are in progress in our lab.