Objective To investigate the expression of microRNA (miRNA)-10a in the intestinal mucosa,serum and peripheral blood mononuclear cell (PBMC) of patients with inflammatory bowel disease (IBD) and explore its role and relevance in the pathogenesis of the disease.Methods The intestinal or colonic mucosal biopsy specimens of nine active ulcerative colitis (UC) patients,11 active Crohn's disease (CD) patients and eight patients with negative colonoscopy result as control were collected.The sera of 12 active UC patients,13 active CD patients and nine healthy controls were collected.The PBMC of nine active UC patients,11 active CD patients and eight healthy controls were collected.The expression of miRNA-10a in the intestinal mucosa,sera and PBMC and the expression of IL-12/IL-23 p40 in the intestinal mucosa were detected by real-time polymerase chain reaction (PCR).Each 8 cases of active UC and CD patients were collected.The intestinal mucosa before infliximab (IFX) treatment and six weeks after three times of IFX treatment were collected.And at same time,the intestinal mucosa of 11 active UC patients and 10 active CD patients were collected and cultured for 18 hours stimulated with IFX in vitro and then the expression of miRNA-10a in the intestinal mucosa was tested.One-way analysis of variance was used for comparison in three samples.Paired t-test was used for two samples comparison.Spearman test was used for correlation analysis.Results Compared with healthy controls,the expression of miRNA-10a in the intestinal mucosa,serum and PBMC of UC and CD patients significantly decreased (F=38.45,30.46 and 14.74,all P＜0.05).There was no statistic significance between UC and CD groups.The expression of IL-12/IL-23 p40 in the intestinal mucosa of UC and CD patients significantly increased (F=32.90,P＜0.05).The expression of IL-12/IL-23 p40 was negatively correlated with the expression of miRNA-10a in the intestinal mucosa of CD patients.After three times of IFX treatment,the expression of miR-10a in the intestinal mucosa of IBD patients significantly increased (t=3.341,3.382,both P＜0.05).After stimulated with IFX in vitro,the expression of miRNA-10a in the intestinal mucosa significantly increased (t=3.095,7.193,both P＜0.05).Conclusions miRNA-10a was closely correlated with the inflammation of IBD patients and with the role of targeting IL-12/IL-23 p40.miRNA-10a might be a new target for the IBD treatment.

Objective To create Mucin gene 1 (MUC1) antisense peptide nucleic acid (PNA),and to observe its effects on MKN-45 cell invasion and explore the mechanism. Methods The sequence of antisense PNA was designed according to MUC1 gene sequence and transfected into human gastric cancer cells (MKN-45) by liposome,and the empty vector group (randomized control group)and blank control group (negative control group) were involved. The expression of MUC1 was detected by real time quantitative PCR and the changes of E-cadherin expression were also observed.The effects on gastric cancer cell invasion were tested with transwell chamber assays.Results The expression of MUC1 gene was effectively suppressed by the 3 created antisense PNA,and their expression level (0.62±0.18,0.49±0.12 and 0.60±0.21) was significantly lower than that of negative control group (1.18 ± 0.03,P ＜ 0.01). There was no significant difference between radomized control group and negative control group (1.00±0.04,P=0.657).After MUC1 PNA transfected,the capability of gastric cancer cell invasion decreased significantly (P=0.005).And the expression of E-cadherin at mRNA and protein level was up-regulated.Conclusions There is negative correlation between MUC1 and E-cadherin expression in gastric cancer cell MKN-45.The capability of tumor cell invasion is significantly inhibited by suppressing MUC1 gene expression.

Cell Transplantation ; adverse effects ; Graft Rejection ; prevention & control ; Hepatocytes ; transplantation ; Humans ; Liver Failure ; surgery

Emerging evidence has demonstrated that genomes are organized into higher-order structures in vivo and long range interactions between genomic regions largely contribute to the regulation of gene expression. Hematopoiesis, orchestrated by the precise spatial regulation and organization of hematopoietic transcription factors, serves as a good model for exploring these issues. The chromosome conformation capture (3C) methodology and its high throughput based technology provide an innovative solution for analyzing the regulation of functional elements through inter-chromosomal and intra-chromosomal interactions, and contacts of functional components in nuclei, thus leading to a more comprehensive understanding of human genome and gene expression. This review focuses on the recent progress of 3C and its derivatives, and their applications in unraveling the mechanisms of transcriptional regulation in hematopoiesis.
Chromosomes ; Gene Expression Regulation ; Genetic Techniques ; Genomics ; Hematopoiesis ; genetics ; Humans ; Nucleic Acid Conformation

Objective: To evaluate the clinical and radiological outcomes,and the complications of staged minimally invasive surgery for adult degenerative scoliosis, comparing with that of the conventional open surgery. Methods: From Jun 2013 to Jun 2017, a total of 42 cases of degenerative scoliosis underwent surgical treatment. Among which, 23 cases underwent staged minimally invasive surgery(MIS group) and 19 cases underwent posterior open surgery(open group). The intra-operative bleeding, and operation time were recorded.The clinical outcomes were assessed using the Oswestry Disability Index (ODI), Visual Analogue Scale (VAS) for low back pain and leg pain.The radiological outcome was evaluated with Cobb angles and sagittal balance parameters (Sagittal Vertical Axis (SVA), Pelvic Incidence (PI)-Lumbar Lordosis (LL), Pelvic Tilt (PT)). The occurrence of complications was recorded. Results: The baseline demographic features of both groups (gender, age, medical comorbidity, etc.) were similar.The mean follow-up period for both groups was more than 2 years.The intraoperative bleeding in the MIS group 405.7±144.8 ml was significantly lower than that in the open group (2 005.3±728.4 ml, t=10.31, P<0.001); The total operation time 414.3±63.0 min of MIS group was significantly longer than that of the open group (304.2±51.8 min, t=6.10, P<0.001), but the operating time of each stage was shorter than the open group. The VAS score for back pain and leg pain,the ODI scores were significantly improved after surgery for both group, there was no significant difference between the two groups, but the low back pain and function in MIS group were better than the open group. The parameters of the coronal and sagittal deformity were significantly improved in both groups, correction of coronal deformity in MIS groupwassignificantly superior to the open group (F=12.02, P=0.001), there was no significant difference in sagittal balance correction between the two groups. The overall complication rate (63.2%) was slightly higher in the open group than in the MIS group (34.8%) without significant difference(χ²=3.36, P=0.07). The incidence of major complications in the open group was significantly higher than that in the MIS group (P=0.014). Conclusion Staged minimally invasive surgery can significantly relieve the patient's pain, improve functionand the coronal and sagittal balance of the patient, the early clinical and radiological outcomes were comparable to the open surgery; Minimally invasive surgery may significantly reduce the amount of bleeding and major complications. Minimally invasive surgery is safe and effective for carefully selected patients with degenerative scoliosis.However, the long-term outcomes of minimally invasive surgery remains to be followed up.

Objective: To evaluate the effect of Danhong injection on endothelial function after percutaneous coronaryintervention (PCI) in coronary heart disease (CHD) . It provides the basis for further research and design. Methods: Sevendatabases of CNKI, VIP, Wanfang Data, CBM, PubMed, The Cochrane Library, Embase and others were searched bycomputer cents with coronary heart disease treated by Danhong injection combined with conventional drugs. Twoevaluators independently sifted the literature, extracted the data and evaluated the bias risk in the study. The data werestatistically analyzed by RevMan 5.3 software. Results: A total of 12 RCTs, involving 1325 patients were included. Metaanalysis showed that the treatment group (Danhong injection combined with routine therapy) was superior to the controlgroup in improving the endothelium index after operation. The index NO[MD=9.57, 95％CI (8.22, 10.93), P ＜ 0.00001], vWF [MD=-31.60, 95％CI (-41.47, -21.72), P ＜ 0.00001], ET-1 [MD=-2.19, 95％CI (-3.11, -1.27), P ＜ 0.00001], ET[SMD=-0.92, 95％CI (-1.49, -0.35), P ＜ 0.01], FMD[MD = 1.81, 95％CI (1.26, 2.37), P ＜ 0.00001]. There was statisticalsignificance between each index. Conclusion: Danhong injection combined with conventional therapy can improveendothelial function after PCI. However, due to the low quality of included studies and the problem of heterogeneity, these conclusions need to be further verified by high quality multicenter, large sample and double blind randomizedcontrolled trials.

OBJECTIVETo investigate the antithrombotic mechanisms of holothurian glycosaminoglycan (GAG) extracted from sea cucumber.

METHODSHuman endothelial cell line EA. hy926 cells were treated with 10 mg/L GAG or 10U/mL unfractionated heparin (UFH) by short-term (15 min - 2 h) and longer-time incubation (6 h - 48 h). Different doses of GAG were used to stimulate EA. hy926. Released free tissue factor pathway inhibitor(TFPI) was determined by ELISA assay. TFPI expression was investigated by immunofluorescent method and TFPI mRNA level by real-time PCR. In a 96-wells microtitre plate, pooled normal plasma containing different concentrations of GAG was allowed to clot by addition of thrombin and calcium chloride, fibrinolysis was induced by addition of t-PA. TRR (TAFI-related retardation of clot lysis) was used to assess thrombin-activatable fibrinolysis inhibitor(TAFI) functional activity.

RESULTSGAG increased TFPI synthesis, expression and secretion in a dose- and time dependent manner. GAG at low concentrations could lengthen while at intermediate concentrations could shorten clot lysis times significantly as compared to control values. TRR was dose-dependently decreased on addition of GAG.

CONCLUSIONSGAG increases TFPI synthesis, expression and secretion of endothelial cells. GAG at intermediate concentrations significantly affects clot stability of a developing clot by means of diminishing TAFI activation.

Animals ; Carboxypeptidase B2 ; antagonists & inhibitors ; Cell Line ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; metabolism ; Glycosaminoglycans ; pharmacology ; Heparin ; pharmacology ; Holothuria ; Humans ; Lipoproteins ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Tissue Extracts ; pharmacology

Adult ; Hemochromatosis ; etiology ; Hepatitis B, Chronic ; complications ; Humans ; Male

OBJECTIVETo screen the mutations of RET proto-oncogene in sporadic patients with pheochromocytoma.

METHODSForty-two cases of sporadic pheochromocytoma were tested for mutations of RET gene. Of these 42 DNA samples, 12 were extracted from peripheral blood cells and 30 from paraffin-embedded pheochromocytoma specimens. The PCR product of exon 10 and exon 11 was used to molecular analysis of the RET proto-oncogene.

RESULTSAmong 42 patients, 2 were found to have RET gene mutations. One of mutations located at codon 634 (TGC>TAC) in exon 11 of RET proto-oncogene. Another one located at codon 632 (GAG>AAG).

CONCLUSIONSome patients with apparently sporadic pheochromacytoma were carrier of mutations, a routine genetic analysis for mutations of RET gene is indicated for these patients.

Adrenal Gland Neoplasms ; diagnosis ; genetics ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Female ; Genetic Predisposition to Disease ; genetics ; Genetic Testing ; Humans ; Male ; Middle Aged ; Mutation ; Pheochromocytoma ; diagnosis ; genetics ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-ret ; genetics

This study was aimed to investigate the effect of GATA-2 over-expression on function of mouse fetal liver hematopoietic stem cells. GATA-2 was introduced into mouse fetal liver cells via retrovirus mediated transduction with GFP as a detecting marker. Flow cytometry, colony-forming assay and cell cycle assay were used to detect the biologic changes of these retrovirus infected mouse fetal liver hematopoietic stem cells. The results showed that GATA-2 over-expression increased the Lin(-)Sca1(+)C-Kit(+) (LSK) population dramatically. Cell cycle of LSK cells didn't show abnormal, while colony forming ability decreased significantly. These data indicated that GATA-2 over-expression inhibited definitive differentiation of mouse fetal liver hematopoietic stem cells. It is concluded that over-expression of GATA-2 can significantly raise the LSK cell proportion in mouse fetal liver and inhibit the differentiation capability, the underlying mechanisms may be related to up-regulation of Hes-1, which may lead to the blocking of cell differentiation at the stem/progenitor cell stage.
Animals ; Cell Differentiation ; Cells, Cultured ; Female ; GATA2 Transcription Factor ; genetics ; Hematopoietic Stem Cells ; cytology ; Liver ; cytology ; Male ; Mice ; Mice, Inbred C57BL