Objective To investigate the relationship between total homocysteine (tHcy) and carotid intima-media thickness (CIMT) in brain infarction patients. Methods Sixty patients with fasting plasma tHcy levels ≤10μmol/L (non-Hhcy group), 60 patients with fasting plasma tHcy levels>10μmol/L and≤15μmol/L (H1 group), and 60 patients with fast-ing plasma tHcy levels>15μmol/L (H2 group) were chosen in hospitalized patients with acute cerebral infarction. Values of CIMT were detected in three groups of patients. The clinical biochemical indicators including triglyceride (TG), total choles-terol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), fasting blood sugar (FBS), folic acid (FA), Vitamin B12 (VitB12) and glycated hemoglobin (HbA1c) were also detected. Results There was signifi-cant difference in CIMT between three groups (P<0.01). The value of CIMT increased in H2 group [0.98(0.90, 1.05)mm] com-pared with that of non-Hhcy group [0.85(0.80, 0.95)mm]. The value of CIMT increased in H2 group compared with H1 group [0.98(0.90, 1.05)mm vs 0.85(0.85, 0.95)mm], P<0.05). There were significant differences in tHcy, FA and VitB12 between three groups. Based on the log-transformed values of CIMT as the dependent variable, multiple stepwise linear regression showed significant associations of the following variables with increased CIMT: increasing age, the history of smoking, the history of diabetes, higher LDL-C and tHcy levels. Conclusion Brain infarction in patients with higher tHcy level often have lower levels of FA and VitB12, and increased CIMT. When the level of tHcy >15 μmol/L, there is more significantly higher level of CIMT. The increased CIMT level was associated with some cerebrovascular risk factors in patients with brain infarction.

Objective To evaluate the efficacy of double filtration plasmapheresis (DFPP) combined with immunosuppressive agents (leflunomide plus methotrexate) on synovitis in magnetic resonance imaging (MRI) in patients with high active rheumatoid arthritis (RA). Methods Fifty eight patients with RA (disease duration 6 months to 12 years) were randomly divided. Thirty-one were randomized to the treatment group and 27 were randomized to the control group. All patients received leflunomide 10 mg, two times daily; plus methotrexate 15 mg orally once weekly. DFPP was performed in the treatment group once 1-2 weeks for 3-4 sessions. Control patients did not receive DFPP. All patients underwent contrast-enhanced MRI of the right wrist at the baseline and 6 months, 1 month in the treatment group. The signs including synovitis pannus, bone marrow edema and effusion were observed on MRI. The scoring of synovial hypertrophy, pannus, bone marrow edema were measured according to the outcome measures in RA MRI scoring system. Comparisons between groups were performed with paired-samples t test and independent-sample t test. Results The MRI synovitis score, MRI pannus score and MRI bone marrow edema in the treatment group was (1.4±1.6), (0.13± 0.35) and (5±4) respectively,so was significantly lower than that of the control group [respectively for (7.9± 1.3), (2.76±0.43), (16±12),P<0.01]. 53% of the treatment group satisfied both the disease activity score 28-joint assessment and MRI synovitis assessment (no enhancement of synovium or pannus, no effusion), but none in the control group (P<0.01). Significant changes at 1 month was observed in DAS28 and HAQ scores (P<0.01), but not in the MRI synovitis score, MRI pannus score, MRI bone marrow edema score and effusion in the treatment group (P>0.05). Conclusion DFPP combined with immunosuppressive agents can significantly improve synovitis in MRI in patients with high active RA. Improvement of the signs of MRI is later than that in the clinic. So imaging assessment may be necessary for accurate evaluation of disease status and selection of therapy.

Objective To evaluate the efficacy of double filtration plasmapheresis (DFPP) in the treatment of patients with refractory rheumatoid arthritis (RA). Methods Eighty-two patients were randomly aesigned,42 to the DFPP group and 40 to the no-DFPP group. All patients previously experienced an incomplete response to 2-3 dis-ease-modifying antirheumatic drugs (DMARDs) and 1-2 nonsteroidal anti-inflammatory drugs (NSAIDs) or predni-sene. All patients received sulphasalazine (SASP,0.75 g three times daily) plus methotrexate (MTX, 10 mg orally once weekly). DFPP was performed once a week for 2-3 sessions. A total of 121 plasmapheresis procedures were per-formed in 42 patients. Control patients did not receive sham DFPP. The efficacy measures recorded one day after the final treatment and latest month in follow up for 12～24 months included the American College of Rheumatology 20% ,50% ,and 70% improvement criteria (ACR20, ACR50, and ACR70), the Health Assessment Questionnaire estimate of disability (HAQ); and the disease activity index. Results Patients in the DFPP group had ACR 20, ACR 50 and ACR 70 improvements of 100% ,92.9% and 81.0%,as compared with the patients in no-DFPP group 17.5% ,0,and 0 (P<0.001). Significant change from baseline was observed in HAQ scores in the DFPP group but not in the no-DFPP group (P<0.001). The changes from baseline in the disease activity scores were significantlygreater than in the no-DFPP group (P<0.001). Conclusion DFPP therapy significantly alters the signs and symp-toms of refractory RA. There are significant increases in physical function and improvement in quality of life.

Objective To study the effect of gonadotropin releasing hormone agonist(GnRHa)against car-boplation-induced gonadotoxicity in rats. Methods Forty Wistar rats were divided into four groups which received carboplation, GnRHa + carboplation, GnRHa and normal saline respectively(n=10 for each group). Blood samples were collected from the abdominal aorta and the levels of blood follicle stimulation hormone (FSH) and estradiol (E<2>) were determined. Both ovaries and uterus of each rat were removed to measure the amount and the maturity of follicles. Body mass and morphological and pathological features of the rats were also observed. Results Compared with that in control group, the body mass of ovary and uterus decreased (P<0.05), and a significant reduction was observed in the number of ovarian follicles at each grade (P<0.05). The levels of E2 significantly lowered (P<0.05) and the level of FSH markedly ascended in group carboplation. Compared with that in group carboplation, the amount of primitive follicles significantly increased in group GnRHa + carboplation (P<0.05), and carboplation showed markedly protective effect on the ovarian and uterine morphological construction of rats. Conclusion Gn-Rha, appliying to preventing the rat reproduction damage in advance, has the certain protective function.

Objective To investigate the cytokines levels in peripheral blood of ulcerative colitis (UC) patients at active stage, and to observe the effect of baicalin at various concentrations on the cytokines. Methods Quantitative polymerase chain reaction (Q-PCR) assay was used for the detection of interleukin 4 receptor (IL4R), IL6R, IL23R and RAR-related orphan receptor C (RORC) expression in single peripheral mononuclear cell of UC patients. Enzyme-linked immunosorbent assay (ELISA) was used to examine the serum levels of interferon gamma (IFN-γ), IL-4, IL-5, IL-6, IL-10, and transforming growth factor beta (TGF-β) levels in UC patients, and was applied for in-vitro detection of these indicators after baicalin intervention. Results Q-PCR results showed that IL4R, IL6R, IL23R, RORC gene expression levels in UC patients were increased to various degrees as compared to diarrhea-predominant irritable bowel syndrome (IBS-D) patients and the healthy volunteers, and then the levels were decreased to various degrees after intervention by baicalin at different concentrations, in particular at 20 and 40μmol/L. The results of ELISA showed that serum levels of cytokines of IFN-γ, IL-5, IL-6 in UC patients were increased to various degrees while IL-4, IL-10 and TGF-β1 was decreased as compared to IBS-D patients and the healthy volunteers. Baicalin at the concentrations of 20 and 40 μmol/L had an effect on decreasing IFN-γ, IL-5, IL-6 and on increasing IL-4 and IL-10. Conclusion Higher concentrations of baicalin show obvious effect on inhibiting RORC and IL23R expression, on decreasing IFN-γ, IL-5, IL-6 levels, and on increasing IL-4, IL-10 and TGF-β1 levels, which indicated that the therapeutic mechanism of baicalin in relieving ulcerative colonic inflammation is related with the regulation of immune function.

Objective To investigate the genotype distribution of microsatellite locus CAI among Candida albicans ( C.albicans ) strains and to evaluate its relationship with the epidemic of vulvovaginal candidiasis ( VVC) in Guizhou region .Methods Ninety independent C.albicans strains isolated from pa-tients with VVC in Guizhou were investigated based on single-strand conformation polymorphisms ( SSCP ) and GeneScan analysis .The genotypes of C.albicans strains were identified by microsatellite locus CAI pol-ymorphism analysis .The gene polymorphism and the cluster of C.albicans strains were analyzed by using software SPSS 19.0.A logistic regression model was used to analyze the relationship between genotype distri -bution of CAI microsatellite among C.albicans strains and VVC infection .Results Twenty-seven distinct CAI genotypes with various patterns were identified from 90 C.albicans strains by GeneScan analysis .Clus-ter analysis showed that the C.albicans strains were classified into three clusters ( ClusterⅠto Cluster Ⅲ) . Three predominant genotypes including 30-45, 32-46 and 30-46 and other 7 highly similar genotypes be-longed to clusterⅡthat accounted for 70.0%(63 strains) in all strains.The odds ratio for the predominant genotypes associated with VVC infection was 4.3.Conclusion The predominant distribution of genotypes was observed among the isolated C.albicans strains.The predominant genotypes of C.albicans were highly associated with the occurrence of VVC .

Background:Tryptophan(TRP)is an essential amino acid,and can produce 5-hydroxytryptamine(5-HT)via 5-HT signal pathway and kynurenine( KYN)metabolic pathway under the catalysis of enzyme,thereby participating in the pathogenesis of visceral hypersensitivity in diarrhea-predominant irritable bowel syndrome(IBS-D). Aims:To investigate the relationship between visceral sensitivity and TRP metabolic pathway in patients with IBS-D. Methods:Thirty patients with IBS-D and 30 healthy controls from June 2012 to January 2014 at Guangdong Provincial TCM Hospital were enrolled. Score of gastrointestinal symptom rating scale( GSRS)was evaluated. Visceral sensitivity was measured by anorectal manometry. RT-PCR and Western blotting were used to detect mRNA and protein expressions of colon mucosal IDo, respectively. Serum 5-HT,5-HIAA,TRP,IDo,KYN,KYNA concentrations and IDo activity,KAT activity were determined by high performance liquid chromatography assay. Results:Compared with control group,GSRS score was significantly increased(P < 0. 05),initial perception threshold,defecation threshold,pain threshold were significantly decreased(P < 0. 05),anorectal constriction pressure was significantly increased( P < 0. 05),serum 5-HT,5-HIAA concentrations were significantly increased(P < 0. 05),mRNA and protein expressions of IDo were significantly increased (P < 0. 05),serum KYN was significantly increased(P < 0. 05),KYNA was significantly decreased(P < 0. 01),IDo activity was significantly incseased(P < 0. 01),and KAT activity was significantly decreased in IBS-D group(P < 0. 01). Correlation analysis showed that initial perception threshold,defecation threshold,pain threshold and anorectal constriction pressure were correlated with 5-HT,5-HIAA,TRP,KYN,KYNA,IDo activity and KAT activity in patients with IBS-D (P < 0. 05). Conclusions:TRP metabolic pathway is correlated with visceral hypersensitivity in patients with IBS-D.

AIM: To detect the effect of conjunction matrigel with mammary fad pat(MFP)implantation on the tumorigenesis, proliferation, apoptosis and metastasis of Her2 positive and negative breast cancer model. METHODS: The Her2 positive BT 474 and Her2 negative MDA-MB 231 breast cancer cells were injected into MFP of nude mice with or without matrigel to establish breast cancer model. The tumor volume was measured every 3 d. Followed up for 30 d after implantation, the nude mice were killed and the tumors and associated organs were dissected for pathological sectioning and staining with hematoxylin and eosin. The time of tumor formation and the tumorigenesis were determined after implantation. The tumor volume and metastasis rate were calculated and compared with each other. The proliferation and apoptosis of Her2 positive and negative tumors were also determined. RESULTS: Matrigel and MFP implantation technology shortened the time of tumorigenesis significantly(P<0.01). The tumorigenesis rate of BT 474 and MDA-MB 231 breast cancer cells did not show any different(P>0.05). The metastasis rate of MDA-MB 231 breast cancer cells were improved from 25.0% to 37.5%(P<0.05). CONCLUSION: Matrigel and MFP implantation can be combined to shorten the time of tumor formation by two kinds of breast cancer cells, and improve the metastasis of Her2 negative MDA-MB 231 cells. Using matrigel does not show any effect of proliferation and apoptosis on Her2 positive and negative breast cancer cells.

Metastasis is a multistep process involving modification of morphology to suit migration, reduction of tumor cell adhesion to the extracellular matrix, increase of cell mobility, tumor cell resistance to anoikis, and other steps. MicroRNAs are well-suited to regulate tumor metastasis due to their capacity to repress numerous target genes in a coordinated manner, thereby enabling their intervention at multiple steps of the invasion-metastasis cascade. In this study, we identified a microRNA exemplifying these attributes, miR-124, whose expression was reduced in aggressive MDA-MB-231 and SK-3rd breast cancer cells. Down-regulation of miR-124 expression in highly aggressive breast cancer cells contributed in part to DNA hypermethylation around the promoters of the three genes encoding miR-124. Ectopic expression of miR-124 in MDA-MB-231 cells suppressed metastasis-related traits including formation of spindle-like morphology, migratory capacity, adhesion to fibronectin, and anoikis. These findings indicate that miR-124 suppresses multiple steps of metastasis by diverse mechanisms in breast cancer cells and suggest a potential application of miR-124 in breast cancer treatment.
Anoikis ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Connective Tissue Growth Factor ; metabolism ; DNA Methylation ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; genetics ; metabolism ; Neoplasm Metastasis ; rho GTP-Binding Proteins ; metabolism ; rho-Associated Kinases ; metabolism

Objective To investigate the effects of small interfering RNA (siRNA) CXCR4 gene on proliferation, invasion and epithelial-mesenchymal transition factors including Twist, Slug and E-cadherin (E-cad)in HeLa cells of human cervical carcinoma.Methods HeLa cells were cultured in vitro and CXCR4-siRNA was transfected into HeLa cells. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the changes of cell proliferation. Transwell experiment was used to observe the changes of cell invasion, and the expressions of CXCR4,Twist,Slug and E-cad protein were detected by Western blot.Results Compared with the blank control group and the negative control group,CXCR4-siRNA transfected into HeLa cells significantly inhibited the expression of CXCR4 protein (0.959±0.197, 0.932±0.141 vs. 0.379±0.022 respectively; F=16.286, P =0.004). MTT results showed that the proliferation of HeLa cells was inhibited at 48 h and 72 h (48 h: 0.846 ± 0.034, 0.823 ± 0.025 vs. 0.744 ± 0.039, F = 7.379, P= 0.024; 72 h: 0.996±0.026, 0.964± 0.059 vs. 0.829±0.051, F = 10.425, P= 0.011). Transwell results showed that the invasiveness of HeLa cells in CXCR4-siRNA group was inhibited at 48 h and 72 h(cell membrane number,48 h:37.4±8.1,33.6±6.2 vs.25.4 ±3.2, F = 4.830, P= 0.029. 72 h: 48.4 ±7.6, 43.0 ±5.4 vs. 33.4 ±6.7, F = 6.579, P= 0.012). After transfection for 48 h, the expression of Twist and Slug protein was decreased and E-cad protein was increased in CXCR4-siRNA group (Twist: 0.93±0.11, 1.00±0.17 vs. 0.53±0.07, F = 10.962, P= 0.010; Slug: 0.515± 0.084,0.470±0.055 vs.0.190±0.028,F =25.579,P=0.001;E-cad:0.53±0.04,0.55±0.24 vs.1.11±0.14,F =12.494, P= 0.007). Conclusions Silencing CXCR4 gene can inhibit the proliferation and reduce the invasiveness of HeLa cells. Inhibiting EMT of HeLa cells can be achieved by down-regulating the expressions of Twist and Slug protein,and up-regulating the expression of E-cad.