Objective To investigate the abnormal expression of microRNA (miRNA)-25 in the serum of gastric cancer patients and its clinical significance. Methods In Xijing Hospital of Digestive Diseases,Fourth Military Medical University,86 gastric cancer patients with operation and completed follow-up data,70 gastric adenoma patients and 80 healthy controls were selected as study objects.Total RNA was isolated from the serum. After the stable and sensitive miRNA-25 absolute quantity detection method established,the serum levels of miRNA-25 in gastric carcinoma patients,gastric adenoma patients and healthy controls were tested according to this method. The expression differences of miRNA-25 in the serum of patients with gastric cancer and gastric adenoma and healthy controls were analyzed with statistic analysis,and the correlation between miRNA-25 expression level and clinic pathological features of gastric cancer was also analyzed. Results The expression level of miRNA-25 in the serum of gastric cancer patients (135. 6 fmol/μg total RNA) was significantly higher than that of gastric adenoma patients (67. 7 fmol/μg total RNA) and healthy controls (62. 2 fmol/μg total RNA)(P＜0. 01). The receiver operating characterisstic curve of miRNA-25 indicated that serum miRNA-25 with good specificity and sensitivity in gastric cancer diagnosis (AUC=0. 827). The serum level of miRNA-25 in gastric cancer patients with lymph node metastasis [(148. 3±10. 2) fmol/μg total RNA] or clinicopathological stage Ⅲ /Ⅳ patients [(146. 7±9.5) fmol/μg total RNA] was significantly higher than that of gastric cancer patients without lymph node metastasis [(120. 3±10. 1)fmol/μg total RNA] or clinicopathological stage Ⅰ/Ⅱpatients [(119. 4±12. 2) fmol/μg total RNA] (P＜0.05). The correlation statistical analysis result indicated that there was no significant difference in survival period between serum miRNA-25 highly expressed and lowly expressed gastric cancer patients (P＞0. 05).Conclusion Serum miRNA-25 testing maybe helpful in diagnosis and prognosis of gastric cancer.

OBJECTIVETo investigate the effect of ketamine on the high-voltage-activated calcium currents (ICa(HVA)) in rat hippocampal neurons.

METHODSNeurons were cultured from Wistar rat hippocampus. ICa(HVA) was recorded using whole-cell patch clamp technique. After application with ketamine at different concentrations (10, 30, 100, 300, and 1000 μmol/L), the effect of ketamine on ICa(HVA) was evaluated.

RESULTSICa(HVA) was inhibited by ketamine in a concentration-dependent manner. Ketamine at 10 μmol/L showed no effect on ICa(HVA). Four concentrations of ketamine (30, 100, 300,and 1000 μmol/L) reduced the peak ICa(HVA) currents by (17.5 ∓ 4.5)%, (25.5 ∓ 6.9)%, (38.5 ∓ 4.1)%, and (42.3 ∓ 4.6)% respectively,with a mean half maximal inhibitory concentration of 68.2 μmol/L and Hill coefficient of 0.47. The maximal activation membrane potential was shifted to (5.3 ∓ 0.8) from (5.4 ∓ 0.9). The half maximal activation membrane potential of inactivation curve was shifted from(-26.7 ∓ 3.9) mV to(-32.8 ∓ 4.2) mV.

CONCLUSIONKetamine can remarkably inhibit calcium currents in the central neurons,which may explain at least partly the action of ketamine on central nervous system.

Animals ; Calcium Channels ; drug effects ; physiology ; Cells, Cultured ; Hippocampus ; drug effects ; physiology ; Ketamine ; pharmacology ; Membrane Potentials ; drug effects ; Neurons ; drug effects ; physiology ; Rats ; Rats, Wistar

OBJECTIVE: To determine the feasibility whether the bovine jugular venous conduit (BJVC) can be fixed with polyepoxy compound (PC). METHODS: Twenty-four BJVCs were divided into 3 groups and fixed with polyepoxy compound (PC group, n = 8), glutaraldehyde (GA group, n = 8), and unfixed group (Control group, n = 8), respectively. The morphologic and mechanical properties of BJVCs in the 3 groups, including thickness, diameter, moisture content, denaturation temperature, tensile strength, elongation at break, and fixation index were measured. The rat subcutaneous model for the assessment of tissue calcification was used. The calcium content in bovine jugular vein patches and valves was determined by flame atomic absorption spectrophotometer. RESULTS: There was no difference in the wall thickness, diameter, and tissue water content between PC and the control group, but significant difference was found between GA and PC groups. The mechanical properties of PC group and GA group were not significantly different, but they were better than those of the control group. GA-fixed BJVC samples showed clear calcification, while PC fixed BJVC were calcified significantly less. CONCLUSION PC is an effective and suitable choice for the treatment of BJVC since it can effectively preserve the structure and the anti-reflow function of valves in bovine jugular vein and it has better anti-calcification properties.
Animals ; Biocompatible Materials ; Bioprosthesis ; Blood Vessel Prosthesis ; Cattle ; Cross-Linking Reagents ; pharmacology ; Epoxy Compounds ; pharmacology ; Jugular Veins ; Polymers

OBJECTIVE: To investigate the effect of decellular treatment on the framework constituents of extracellular matrix and tissue stability in bovine jugular vein conduit (BJVC), and to provide an evidence for tissue engineering of vascular prosthesis. METHODS: Bovine jugular veins were obtained fresh from a local slaughterhouse and were stored in chilled PBS. In the laboratory, any fat and loose connective tissue on the outer surface of the vessel was trimmed. BJVCs were decellularized by a 3-step extraction method as detergent Triton X-100 (0.5%), Trypsin (0.025%) EDTA (0.02%), and DNase I(30kU/L) RNaseA(0.3g/L). Histological and transmission electron microscopy (TEM) techniques were used to study the framework constituents of extracellular matrix of treated the examples, and fresh tissues were used as controls. Tissue contents of hydroxyproline(alkaline hydrolysis method) and elastin (Fastin Elastin Assay) were assayed respectively in the fresh and decellularized groups (n=10). The vascular wall heat shrinking temperature and mechanical strength were measured to evaluate the tissue stability (n=10). RESULTS: Histochemical and TEM analysis of BJVCs treated with decellularization proved a complete removal of nuclear and other cell components. Tissue collagen was well kept,but elastin was partly lessened. Tissue content of hydroxyproline increased comparatively [(25.73+/-2.97)mg/g vs. (29.25+/-2.99)mg/g, P<0.05] and the elastin content obviously decreased [(159.71+/-21.06)mg/g vs. (134.91+/-35.40)mg/g, P<0.05] in the decellular treatment group compared with the control group. The heat shrinking temperature and tensile stress of decelluarized tissue were lower than those of the fresh tissue[(72.50+/-0.53) degrees C vs. (69.75+/-0.54)degrees C ,P<0.05], [(5.19+/-0.65)MPa vs. (3.13+/-0.94)MPa, P<0.05]. CONCLUSION The basic framework of extracellular matrix in the decellularized BJVC is partly damaged and tissue stability is reduced. Decellularized BJVC should be further crosslinked before being used as a tissue engineering scaffold for clinical pulmonary artery graft.
Animals ; Blood Vessel Prosthesis ; Cattle ; Extracellular Matrix ; Jugular Veins ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo promote stem cells differentiation into neurons and enhance neuromotor function after brain injury through brain-derived neurotrophic factor (BDNF) induction.

METHODSRecombinant adenovirus vector was applied to the transfection of BDNF into human-derived umbilical cord mesenchymal stem cells (UCMSCs). Enzyme linked immunosorbent assay (ELISA) was used to determine the secretion phase of BDNF. The brain injury model of athymic mice induced by hydraulic pressure percussion was established for transplantation of stem cells into the edge of injury site. Nerve function scores were obtained, and the expression level of transfected and non-transfected BDNF, proportion of neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP), and the number of apoptosis cells were compared respectively.

RESULTSThe BDNF expression achieved its stabilization at a high level 72 hours after gene transfection. The mouse obtained a better score of nerve function, and the proportion of the NSE-positive cells increased significantly (P<0.05), but GFAP-positive cells decreased in BDNF-UCMSCs group compared with the other two groups (P<0.05). At the site of high expression of BDNF, the number of apoptosis cells decreased markedly.

CONCLUSIONBDNF gene can promote the differentiation of the stem cells into neurons rather than glial cells, and enhance neuromotor function after brain injury.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Brain Injuries ; physiopathology ; therapy ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Mice ; Nerve Tissue Proteins ; analysis ; Neurons ; cytology ; Phosphopyruvate Hydratase ; analysis ; Transfection

OBJECTIVETo investigate the safety and efficacy of intracoronary transplantation of G-CSF mobilized autologous peripheral blood stem cells in patients with acute myocardial infarction (AMI).

METHODSPatients with AMI were randomly assigned to receive intracoronary PBSCs transplantation following bone marrow cells mobilization by granulocyte colony-stimulating factor (300-600 microg/day subcutaneously for 5 days) in addition to standard therapy (standard drug therapy and PCI, PBSCs transplantation group, n = 35) or standard therapy (standard drug therapy and PCI, n = 35). One day after G-CSF treatment was finished the patient's mononuclear cells were harvested by Baxter CS 3000 blood cell separator in a volume of 57 ml and then transferred into the infarct related artery by occluding the over the wire balloon and infusing artery through balloon center lumen. Complications during intervention and left ventricular function at baseline and 6 months thereafter were monitored.

RESULTSNo severe side effects of G-CSF treatment could be observed. Malignant arrhythmias were not observed either. Left ventricular function was significantly improved 6 months after G-CSF mobilized autologous peripheral blood stem cell transplantation compared to baseline (global left ventricular function ejection fraction: 57.1 +/- 7.8 vs. 50.0 +/- 8.2%, P < 0.0001; WMSI: 1.101 +/- 0.118 vs. 1.219 +/- 0.190, P < 0.0001; left end-systolic volume: 52.6 +/- 20.3 vs. 63.8 +/- 23.9 ml, P = 0.01 and left end-diastolic volume: 119.2 +/- 30.3 vs. 134.2 +/- 36.7 ml, P = 0.07) while these parameters remained unchanged in the control group.

CONCLUSIONThe present study demonstrates that G-CSF mobilized autologous intracoronary PBSCs transplantation is a safe and feasible treatment for patients with AMI and global left ventricular function is improved and left ventricular remodeling attenuated at six-month follow-up.

Aged ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; surgery ; therapy ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous ; Treatment Outcome

Objective To learn the spatial and temporal patterns of primary syphilis and secondary syphilis in Shenzhen and to provide evidence for carrying out further research on syphilis.Methods Primary syphilis and secondary syphilis cases among residents in Shenzhen between 2005and 2009(n=11 303) were geocoded at street office level (n=55) based on residence at the time of diagnosis. Both spatial and space-time scan statistics were used to identify clusters of street office by using SaTScan software. Results In the purely spatial analyses, clusters were seen in the junction of the Baoan district and Nanshan district (Xinan, Xixiang, Nanshan and Nantou street office) and in the region near Hong Kong (Dongmen, Shekou, and Futian street office), as well as in the other streets where entertainment industry was relatively developed (Longhua, Huafu, Huangbei and Cuizu street office). The clusters had not changed much in the first four years, but nine clusters appeared in 2009.Annually, the most likely clusters were located in Longhua (2005, P≤0.001, RR=3.34), Bamboo (2006, P≤0.001, RR=9.59), Huafu (2007, 2008 years, P≤0.001, RR values were 4.18 and 4.75)and Cuizu (2009, P≤0.001, RR=8.02). In the space-time scan analysis, we found 16 significant clusters, which were similar to the pure spatial analyses. However, regional difference were also found, with the most likely cluster was the Guiyuan street office in 2006. Conclusion Spatial and space-time scan statistics seemed to be effective ways in describing the circular disease clusters. We have had a better understanding on spatial and temporal patterns of primary syphilis and secondary syphilis in Shenzhen through spatial and space-time scan statistics of syphilis surveillance data in the recent years. The changes of spatial and temporal patterns of primary syphilis and secondary syphilis were also described by SaTScan software, which also provided useful reference for the preventive strategies on sexually transmitted diseases as well as on HIV. Useful information was also provided for financial investment and cost-effective studies.

Objective: To evaluate clinical characteristics and prognosis of primary myelofibrosis (PMF) patients with thrombocytopenia in varied degrees. Methods: Clinical features and survival data of 1 305 Chinese patients with PMF were retrospectively analyzed. The prognostic value of thrombocytopenia in patients with PMF was evaluated. Results: 320 subjects (47%) presented severe thrombocytopenia (PLT<50×10(9)/L), 198 ones (15.2%) mild thrombocytopenia [PLT (50-99)×10(9)/L] and 787 ones (60.3%) without thrombocytopenia (PLT ≥ 100×10(9)/L). The more severe the thrombocytopenia, the higher the proportions of HGB<100 g/L, WBC<4×10(9)/L, circulating blasts ≥ 3%, abnormal karyotype and unfavourable cytogenetics (P<0.001, P<0.001, P=0.004, P<0.001 and P<0.001, respectively) were observed in this cohort of patients. The more severe the thrombocytopenia, the lower the proportion of JAK2V617F positive (P<0.001) was also noticed. Platelet count was positively correlated with splenomegaly, HGB and WBC (P<0.001, correlation coefficients were 0.131, 0.445 and 0.156, respectively). Platelet count was negative correlated with constitutional symptoms and circulating blasts (P=0.009, P=0.045, respectively; correlation coefficients were -0.096 and -0.056, respectively). The median survival of patients with severe thrombocytopenia, mild thrombocytopenia and without thrombocytopenia were 32, 67 and 89 months, respectively (P<0.001). Multivariate analysis identified thrombocytopenia in varied degrees (HR=1.693, 95%CI 1.320-2.173, P<0.001) and Dynamic Internation Prognostic Scoring System(DIPSS) prognostic model (HR=2.051, 95%CI 1.511-2.784, P<0.001) as independent risk factors for survival. Conclusion: PMF patients with severe thrombocytopenia frequently displayed anemia, leucopenia, circulating blasts and short survival, so active treatment measures should be taken especially in these patients.
Humans ; Primary Myelofibrosis ; Prognosis ; Retrospective Studies ; Thrombocytopenia

Objective: To explore the clinical implications and prognostic value of TP53 gene mutation and deletion in patients with myelodysplastic syndromes (MDS) . Methods: 112-gene targeted sequencing and interphase fluorescence in situ hybridization (FISH) were used to detect TP53 mutation and deletion in 584 patients with newly diagnosed primary MDS who were admitted from October 2009 to December 2017. The association of TP53 mutation and deletion with several clinical features and their prognostic significance were analyzed. Results: Alterations in TP53 were found in 42 (7.2%) cases. Of these, 31 (5.3%) cases showed TP53 mutation only, 8 (1.4%) cases in TP53 deletion only, 3 (0.5%) cases harboring both mutation and deletion. A total of 37 mutations were detected in 34 patients, most of them (94.6%) were located in the DNA binding domain (exon5-8) , the remaining 2 were located in exon 10 and splice site respectively. Patients with TP53 alterations harbored significantly more mutations than whom without alterations (z=-2.418, P=0.016) . The median age of patients with TP53 alterations was higher than their counterparts[60 (21-78) years old vs 52 (14-83) years old, z=-2.188, P=0.029]. TP53 alterations correlated with complex karyotype and International prognostic scoring system intermediate-2/high significantly (P<0.001) . Median overall survival of patients with TP53 alterations was shorter than the others[13 (95%CI 7.57-18.43) months vs not reached, χ(2)=12.342, P<0.001], while the significance was lost during complex karyotype adjusted analysis in multivariable model. Conclusion: TP53 mutation was more common than deletion in MDS patients. The majority of mutations were located in the DNA binding domain. TP53 alterations were strongly associated with complex karyotype and always coexisted with other gene mutations. TP53 alteration was no longer an independent prognostic factor when complex karyotype were occurred in MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Genes, p53 ; Humans ; In Situ Hybridization, Fluorescence ; Middle Aged ; Mutation ; Myelodysplastic Syndromes/genetics* ; Prognosis ; Tumor Suppressor Protein p53 ; Young Adult

OBJECTIVETo investigate the association of single nucleus polymorphisms(SNP)of tumor necrosis factor alpha (TNF-α) gene (-308 G>A and -238 G>A genotypes) with susceptibility to primary myelodysplastic syndromes (MDS).

METHODSTwo SNPs (TNF-α-308 G>A,TNF-α-238 G>A) of TNF-α gene were detected by Taqman probes in 341 MDS patients and 365 unrelated-healthy controls.

RESULTSCompared to healthy controls, the frequency of TNF-α-308 AA+AG genotype and A allele increased (18% vs 10%, P=0.015, 9% vs 5%, P=0.021, respectively) in refractory cytopenia with multilineage dysplasia (RCMD) patients. There was no correlation of TNF-α-308 G>A genotype and allele frequency between MDS and controls. No difference in the genotype and allele frequency of TNF-α-238 G>A were found between controls and MDS or the subtypes of MDS (P>0.05). We did not find any linkage between plasma level of TNF-α and TNF-α-308 G>A or TNF-α-238 G>A genotype. Statistic differences were observed between platelet count[58(1-611)×10⁹/L vs 90(7-352)×10⁹/L]and bone marrow blasts in MDS patients carrying TNF-α-308 G>A GG and AA+AG genotype (P=0.024, 0.019, respectively).

CONCLUSIONTNF-α-308 G>A polymorphism was correlated with susceptibility to MDS-RCMD.

Case-Control Studies ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Myelodysplastic Syndromes ; genetics ; Polymorphism, Single Nucleotide ; Tumor Necrosis Factor-alpha ; blood ; genetics