Objective To study the association of SNP of the AdipoR1 gene with T2DM in Xi’an population. Methods The amplification refractory mutation system(ARMS) analysis and gene sequencing were used to investigate the AdipoR1 gene polymorphism in 100 type 2 diabetics and 84 normal control subjects. Results (1) The genotype and allele frequencies of -106A/G, 5843A/G were not significantly different between type 2 diabetics and normal control subjects. (2) The diagnosis age of diabetics was significantly younger in AdipoR1 5843GG genotype group than in other genotype groups. Conclusions The data implicate that the AdipoR1 gene -106A/G and 5843A/G polymorphism may be not associated with pathogenesis of T2DM. AdipoR1 5843 GG genotype may be associated with the earlier diagnosis of T2DM

Objective To study the change of serum nitric oxide level in patients with type 2 diabetes (2-DM) and in patients with diabetic peripheral neurology (DPN). Methods Nitrose reductase method was used to examine the serum concentration of NO in type 2 diabetics (n=74) and those accompanied with DPN (n=22) and without DPN (n=21). Results ① The serum NO level in the early stage of metabolic disorder of 2-DM patients was higher than that in the control group; it showed a declining trend in the process of DM and was significantly lower in the middle and late stages of DM. ② The 2-DM patients with DPN had significantly lower serum NO level than in those without DPN, but no difference was observed in serum NO level between the diabetics with mild DPN and severe DPN. Conclusion ① Serum NO level in 2-DM patients has a descending changes in the progression of the disease. ② Serum NO level is closely correlated with the occurrence of DPN, but has no obvious association with its progression.

Marrow stromal cells ( MSCs) , component of the mar-row stroma, play an important role in the growth of bone and metabolic balance. This paper mainly summarizes and reviews the molecular regulaory effects of vitamin D and its active form 1, 25 dihydroxy vitamin D3[1,25(OH)2D3] on the osteogenet-ic differentiation of MSCs by Wnt signaling pathway, Wnt5a/ROR2 axis, BMP/TGF-β/Samd signaling pathway and ROS/ERK signaling pathway, so as to clarify the molecular signaling pathway through which vitamin D regulates MSCs metabolically.

Objective To separate and screen components with fibrinolytic enzyme activity from twelve Chinese herbal medicines. Methods The components were extracted with water and precipitated with salt, and they were tested by fibrinolytic protein plates method. The active components with fibrinolytic activity were separated and screened which were compared with urokinase. Results Eleven of the twelve extracts showed fibrinolytic activity, while Trichosanthes kirilowii got the biggest fibrinolytic zone after 36 hours, followed by Alisma plantago-aquatica and Leonurus japonicus, and the Radix Astagali got the smallest one. According to the concentration of the protein, the area of the fibrinolytic zone and the specific activity of the components, the extract from Angelica sinensis exhibited the best specific activity at level of 48.46U/mg. Conclusion The extracts from Chinese herbal medicines except Semen Persicae exhibit fibrinolytic enzyme activity which can dissolve the fibrin in different degrees.

AIM: To investigate the immunoblot changeS of dystrophin expression in mdx mice after bone marrow stem cells transplantation. METHODS: The experiment was conducted in the laboratory of Department of Neurology, the First Affiliated Hospital, Sun Yat-sen University from September to December 2004. ①Twenty-five mdx mice of 7-8 weeks old were selected and randomly divided into transplantation 4, 8, 12 and 16 weeks groups and blank control group with 5 mice in each group. Meanwhile, 20 C57 mice of 4 to 6 weeks old were selected as donor mice, and 5 C57 mice aged 10 weeks served as positive control mice. ②All mice were preconditioned with 7 Gy ?-ray. After radiotherapy, 2?107 marrow stem cells per mouse (about 0.3-0.5 mL) were injected into the vena caudalis of mice; 0.3 mL PBS was injected into the blank control group and positive control group. ③The mice in 4 transplantation groups were killed at each time point, and the gastrocnemius was harvested to prepare dystrophin samples. The amount of dystrophin expression was detected by Western blot with GAPDH as control. RESULTS: All 25 mdx mice and 5 C57 mice were involved in the result analysis. Western Blot analysis of dystrophin after transplantation: No dystrophin was detected in the blank control group; in the 4 weeks after transplantation group, only few dystrophin expressions were detected, and the dystrophin/GAPDH was about 0.095?0.267; in the 12 weeks after transplantation group, dystrophin/GAPDH was about 0.218?0.338; dystrophin expressions were increased with time, at 16 weeks after transplantation, the expressions were much more than those at 8 weeks (dystrophin/GAPDH: 0.393?0.385, 0.173?0.284, t =6.062, P

Amyotrophic lateral sclerosis is a progressive neurodegenerative disease that targets motor neurons without efficient treatment. Stem cell receives the enormous attention because it from specific tissues can differentiate into motor neuron. Stem cells from specific tissues could be used for amyotrophic lateral sclerosis, such as embryonic stem cells, neural stem cells, bone marrow mesenchymal stem cells, umbilical cord blood-derived stem cells and induced pluripotent stem cells. The mechanism of stem-cell therapy includes cell-replacement, delivery neurotrophic factors and immunomodulation. Animal researches and some clinical trails have confirmed that stem cells have great potential to treat neurodegenerative diseases. Till now, people are still unknown some aspects of stem cells, and many problems still need to be resolved.

Objective:To observe the effects of All-trans retinoic acid (ATRA) on the immune functions of AChR-specific lymphcytes via in vitro assays,and investigate the possibility of ATRA in the clinical treatment of myasthenia gravis (MG).Methods:CFA control group and EAMG experimental rats were established to obtain single lymphocytes suspension and cells were followed by AChR97-116 peptide with or without ATRA stimulation for 72 h,and then viable cell population,cell apoptosis,cell cycle and the distribution of Th cells were determined by flow cytometry.CCK-8 assay was selected to evaluate the effects of ATRA on proliferatory ability of lymphocytes.ELISA was used to detect the antibody secretion of B cells affected by ATRA.Results:Compared with CFA group,lymphocytes obtained from EAMG rats had higher ratios of living cells,and this ratio was obviously decreased after ATRA treatment,P＜0.001.Different concentrations of ATRA promoted the apoptosis of AChR-specific cells (P＜0.001),and the promoted effects were ATRA dose-dependent,however,cell cycles were not changed.ATRA markedly inhibited the proliferation of cells from both CFA and EAMG groups,moreover,AChR-specific cells were more sensitive to ATRA treatment (P＜0.01) than that of cells from CFA rats (P＜0.05).The ratio of AChR-specific CD4+T cells was reduced by ATRA (P＜0.01),and ATRA incubation significantly promoted the percentages of Th2,(PCD4+-4IL-4+＜0.001),Treg (PCD4+-Foxp3+＜0.001) cell types,but markedly inhibited the percentages ofThl7 (PCD4+-IL-17+＜0.05),Thl (PCD4+-IFN-γ+＜0.001) cells.ELISA data showed us that ATRA obviously down regulated the antibody secretion of AChR-specific B cells,P＜0.01.Conclusions:ATRA not only inhibited the functions of AChR-specific T cells,but also suppressed the roles of AChR-specific B cells,predicating a therapeutic effect of ATRA on myasthenia gravis therapy.

Objective To investigate the effect of cervical knife conization (CKC) or loop electrical excision procedure (LEEP)on the outcome of subsequent pregnancies and mode of deliveries. Methods A retrospective case-control study including 228 women after treatment with LEEP or CKC for cervical intraepithelial neoplasia (CIN) Ⅱ -Ⅲ who gave birth in the First Affiliated Hospital of Sun Yat-sen University and He-xian Memorial Hospital of Pangyu from January 2004 to January 2010 was performed.Patients (n = 228) without cervical surgical history were randomly extracted from the respective hospitals birth registries as controls and were matched by age, gestation,parity and income.The information including gestational age, premature rupture of membranes (PROM), type of deliveries and birth weight of the two groups were collected.Results The gestational age of women treated with conization was (268.3±26.2) d, longer than that of the women without surgery (279.4±25.3) d (t=4.60, P＜0.01). The incidence of preterm birth was 18.0％(41/228) and 4.4％ (10/228) (x2 = 21.22, P＜ 0. 05). The incidence of PROM was higher in conizationgroup (10.1％, 23/228) than that (1.3％, 3/228) in control group (x2=16.32, P＜0. 05). Risk for PROM was almost eight fold (OR=8. 42, 95％CI: 2.49-28.44) higher in conization group. Cesarean section rate was higher in conization group (69.3 ％ ) than in control group (39.0 ％ )(x2=42.06, P＜0. 01). The gestational age of women treated with LEEP was longer than those treated with CKC[(269.8±24.6) d vs (260.2± 26.5) d, t= 4. 01, P＜0.01]. The incidence of preterm birth was 13. 1％ (22/168) and 31.6％ (19/60) (x2 = 10. 34, P＜0. 05). The mean birth weight of women with LEEP was heavier than that with CKC[(3358.5 ±812.2) g vs (3295.9 ±832.6) g, t=3.08, P＜0. 01]. The incidence of PROM (7.1％, 12/168) of woman with CKC was higher than that (1.3％, 11/60) of women with LEEP (x2 =6.10, P＜0.05). Conclusions Conization might increase the incidence of preterm delivery and preterm PROM. LEEP showed less adverse effect onthe outcome of subsequentpregnanciesthan CKC,and waspreferredfor primigravida, and the risk of treatment should be informed in advance.

Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.

Objective To observe the expression of MMP-1, TNF-a in cholesteatoma and to determine their roles in the destruction of bone and their correlation. Methods Immunohistochemical method and the computer image quantitative analysis were used to examine the expression of TNF-α and MMP-1 in 22 cases of chotesteatomamiddle ear and 20 cases of normal external acoustic meatus skin. Results Positive stainings of MMP-1 and TNF-α were both localized in cytoplasm. The MMP-1 positive cells were found in all strata of cholesteatoma epithelium and active multiplication stromal cell. TNF-α was expressed in both epithlium and stromal cells. The results of the computer image quantitative analysis showed that the mean optical density of MMP-1 (0. 2013±0. 0106) and TNF-α (0.3852±0.0318) in cholesteatoma were higher than that in normal skin epithelial tissue( P＜0.05 ). Conclusion (1)MMP-1 and TNF-α are overexpressed in cholesteatoma. (2)MMP-1 and TNF-α have a correlation in their expression. (3)MMP-1 and TNF-α are both observed in stromal cells which indicates that stromal cells play an irnportant role in bone destruction.