Aim To observe the change of amyloid, acetylcholine transferase and glial fibrillary acidic protein expression in Macaca Rhesus hippocampal after infused the Aβ_(1-42) and thiorphan and explore the possibility of the establishment of Macaca Rhsus AD model in brain.Method The Rhesus monkeys were anesthetized (im), the skull was exposed by a midline scalp incision, and oriented craniotomy was performed on left side by dental drill.First, neprilysin in cerebral cortex and basal nucleus was consumed by infusion thiorphan. Then cerebral cortex and basal nucleus were slowly infused with fibrilla Aβ_(1-42). Finally, the cannula for thiorphan infusion was implanted into the basal nucleus.Miniosmotic pump (Alzet MODEL 2ML4,) was subcutaneously fixed by bio gel 454 on the calvaria (Loctite Co. Ltd,USA) according to the manufacturer's instructions.After 50 days' survival, animals were deep anesthetized with ketamine and sacrificed. The pathological changes were observed by HE staining and immunostaining in monkey brains.Result Neuronal loss and a proliferation of microglia were detected in hippocampal formation by HE staining.Immuno-staining showed Aβ_(1-42),ChAT and GFAP positive cells density were 0.59±0.05,0.21±0.04 and 0.19±0.04 separately.Compared with control group, the density in experimental groups showed distinct difference in statistic analysis (P<0.01).Conclusion The same pathological change was detected in the thioaphan and Aβ_(1-42) infusion in Macaca Rhesus hippocampal formation as what was found in AD patients.

To investigate the clinicopathological features of endometrial endometrioid adenocarcinoma(EEA)with adnexal involvement.Methods The clinicopathological data of 86 EEA patients who underwent surgical treatment at our center from January 2000 to December 2015 were analyzed retrospectively.The clinicopathological features were compared between patients with occult adnexal involvement and those with gross adnexal involvement.Results A total of 86 EEA patients with adnexal involvement(mean age:58.1 years)were included in this study,accounting for 5.4%(86/1592)of the EEA patients during the same period.Among these 86 patients,there were 13 premenopausal patients(15.1%)including 2 premenopausal patients aged under 40 years.Gross adnexal involvement was found in 47 patients(54.7%),while occult adnexal involvement was found in 39 patients(45.3%)in pathology evaluation.Ovarian metastasis was found in 34 patients(39.5%),followed by both ovarian and tubal metastasis in 19 patients(22.1%)and tubal metastasis in 33 patients(38.4%).The expressionss of estrogen receptor(χ=8.086,P=0.042)and progesterone receptor(PR)(χ=9.149,P=0.026)were significantly different between gross adnexal involvement group and occult adnexal involvement group,whereas no significant difference was found in other clinicopathological features(all P>0.05).The non-conditional Logistic regression analysis showed that,compared with PR no-expression group,the rate of occult microscopic adnexal involvement in PR low-expression group was 6.375 times of that of the gross adnexal involvement(P=0.005,95%CI:1.768-22.976),and the rate of occult microscopic adnexal involvement in the PR high-expression group was 3.719 times of that of gross adnexal involvement(P=0.048,95%CI:1.009-13.702). Conclusion PR expression level is remarkably lower in EEA patients with gross adnexal involvement than those with occult adenxal involvement.

OBJECTIVETo investigate the relationship between the resistance index (RI) of the renal artery and serum creatinine (Cr) level in patients early (within one month) after renal transplantation.

METHODSA total of 123 patients receiving renal transplantation underwent examinations by color Doppler ultrasound for measurement of the RI of the renal artery within one month after the operation. According to the results of RI measurement, the patients were divided into RI≥0.75 and RI<0.75 groups for analyzing the correlation between RI and serum Cr level measured at the same time points.

RESULTSThe RI and Cr levels in patients with RI≥0.75 showed a significant positive correlation (P<0.05), whereas they showed an inverse correlation in patients with RI<0.75 (P<0.05). The patients with RI≥0.75 had significantly lower RI of the renal artery and Cr level than those with RI≥0.75.

CONCLUSIONRI is significantly correlated to Cr, and may serve as an indicator for predicting renal graft function after transplantation.

Adolescent ; Adult ; Aged ; Creatinine ; blood ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Postoperative Period ; Renal Artery ; diagnostic imaging ; physiopathology ; Ultrasonography, Doppler ; Vascular Resistance ; Young Adult

Objective To investigate the effects of cognitive interventions (CIs) in the context of communi- ty based rehabilitation (CBR) on cognitive deficits (CDs) in stroke patients.Methods Ninety-two stoke patients with CDs were randomly divided into a CI group and a control group.All patients were treated with conventional CBR.In addition,the patients in the CI group were also treated with special intervention therapy.The patients in both groups were assessed with the neurological and cognitive status examination (NCSE) for cognitive functioning, the FCA for motor function and the BI for their ability in the activities of daily living.Results The NCSE,FCA and BI scores in the cognitive intervention group after treatment were significantly higher than those before treatment and also significantly higher than those in the control group after treatment.Conclusion CIs can not only improve CDs,but also enhance recovery of motor function and ADL.

The neuroimaging results of drug-resistant epilepsy patients play an important role in the surgery decision and prognosis. The aim of this study was to evaluate the impact of these results on the efficacy of epilepay surgery, and then to explore surgical benefit for epilepsy patients with negative magnetic resonance (MR) images. Twenty-four subgroups describing the outcomes of 1475 epilepsy patients with positive-neuroimaging results and 696 patients with negative-neuroimaging results were involved in the meta-analysis. Overall, the odds of postoperational seizure-free rate were 2.03 times higher in magnetic resonance imaging-positive (MRI-positive) patients than in MRI-negative patients [odds ratio (OR)=2.03, 95% CI (1.67, 2.47), P<0.00001]. For patients with temporal lobe epilepsy (TLE), the odds were 1.76 times higher in those with MRI-positive results than in those with MRI-negative results [OR=1.76, 95% CI (1.34, 2.32), P<0.0001]. For patients with extra-temporal lobe epilepsy (extra-TLE), the odds were 2.88 times higher in MRI-positive patients than in MRI-negative patients [OR=2.88, 95% CI (1.53, 5.43), P=0.001]. It was concluded that the seizure-free rate of MRI-positive patients after surgery was higher than that of MRI-negative patients. For patients with negative results, an appropriate surgery should be concerned for TLE.
China ; epidemiology ; Epilepsy ; diagnosis ; epidemiology ; surgery ; Humans ; Magnetic Resonance Imaging ; statistics & numerical data ; Neurosurgical Procedures ; statistics & numerical data ; Prevalence ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; Surgery, Computer-Assisted ; statistics & numerical data ; Treatment Outcome

OBJECTIVE: To compare the flow-mediated dilatation (FMD) among the newly diagnosed impaired glucose tolerance (IGT), type 2 diabetes mellitus (T2DM), and the normal controls (NC) and to analyze relevant factors under different glucose levels. METHODS: The study included IGT (n=34), DM1 (n=52), DM2 (n=33) and NC (n=25). Levels of fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (PPG), fasting insulin (FINS), 2-hour postprandial insulin (PINS), triglyceride (TG), total cholesterol (TC), and hemoglobin A1C (HbA1C) were determined in all participants. High resolution ultrasound examining FMD was performed to measure vascular endothelial function subsequently. RESULTS: There was statistically significant difference between IGT, DM, HG and NC group in FMD (P=0.008). Partial correlation analysis found that a significant negative correlation existed between FMD and homeostasis model assessment-index (HOMA-IRI), difference of plasma glucose (DPG), FPG and PPG (P<0.01), and a negative correlation between FMD and HbA1C (P<0.05). Setting FMD as dependent variable to conduct multiple linear stepwise regression, in IGT group it was the waist/hip ratio (WHR) and HOMA-IRI that entered the regression equation; in DM1 group it was HOMA-IRI, PPG and DPG that entered the regression equation; in DM2 group it was FPG and HOMA-beta that entered the regression equation. CONCLUSION There exists a flow-mediated vasodilatation dysfunction in patients of newly diagnosed IGT and T2DM. Effect of relevant factors on FMD differs with different glucose levels.
Adult ; Blood Glucose ; metabolism ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; blood ; physiopathology ; Endothelium, Vascular ; physiopathology ; Female ; Glucose Intolerance ; blood ; physiopathology ; Humans ; Male ; Middle Aged

Enhanced green fluorescent protein( EGFP), myc epitope and polyhistidine metal-binding tag are often used as a marker for recombinant fusion protein in many gene expression vectors, each marker has its own function, EGFP emits green fluorescence for direct detection, myc epitope facilitates recombinant fusion protein detection using its antibodies, polyhistidine tag allows purification of recombinant fusion protein using resin.Hitherto, no a plasmid vector can integrate all of these functions. In this study we constructed a novel eukaryotic expressive plasmid, designated as pcDNA6/myc-his-EGFP B, which integrated the functions of EGFP, myc epitope and polyhistidine tag. Importantly, a linker octo - peptide in N terminal of EGFP was designed using LINKER program. A DNA fragment encoding a putative protein containing a signal peptide of human interleukin 2(IL-2) in N terminal was cloned into pcDNA6/myc-his-EGFP B in frame with the C-terminal peptide to construct pMHES. 2.2.15 Cells were transfected with pcDNA6/myc-his-EGFP B and pMHES, and Balb/c mice were intravenously injected with pcDNA6/myc-his-EGFP B by tails, results revealed that both of the plasmids worked in 2.2.15 Cells and livers of Balb/c mice. Assuming gene of the IL-2 was inserted into pcDNA6/myc-his -EGFP B in frame with EGFP, myc and 6 × His, three-dimensional structure for this putative expression product was simulated using Modeller8V2, results revealed that IL-2, EGFP, myc and 6 × his did not interfere each other and octo- peptide linker owned certain flexibility. The results suggest that pcDNA6/myc-his-EGFP B may be useful as a genetic tool for mammalian cells and a vector for gene therapy.

OBJECTIVETo investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.

METHODSHUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.

RESULTSs The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.

CONCLUSIONDigestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.

Cell Culture Techniques ; Cell Cycle ; Cells, Cultured ; Culture Media ; chemistry ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 8 ; chemistry ; Serum

Objective To investigate the characteristics of gut microbiota in patients with schizophrenia. Methods The composition of the fecal microbiota in 16 schizophrenia patients and 20 healthy controls was analyzed by 16S rRNA sequencing. Results At the phylum level, TM7 was increased in the patients (P<0.05). At the class level, the abundance of Peptostreptococcaceae, Lactobacillaceae and Actinomycetaceae in the patients increased (P<0.05), whereas the abundance of Tissierellaceae decreased relative to health controls (P<0.05). At the genus level, Actinomyces, Scardovia, Atopobium, Moryella, G07 and Lactobacillus were increased in the patients relative to healthy controls (P<0.05). Prediction of functional properties of bacterial flora showed that the metabolic pathways of fatty acids and pyruvate were different between the patients and healthy controls (P<0.05). Conclusion The present study reveals an alteration in the characteristics of gut microbiota in schizophrenia patients which may be an auxiliary diagnosis marker for clinical diagnosis of schizophrenia.

OBJECTIVEMultidrug resistance (MDR) is one of the primary causes of suboptimal outcomes in chemotherapy of children with acute myeloblastic leukemia (AML). The mechanisms of drug transport resistance may chiefly contribute to MDR. Expression and/or activity of P-glycoprotein (P-gp), multiple resistance-associated protein-1 (MRP1), lung-resistance related protein (LRP) and breast cancer resistance protein (BCRP) have been considered to be associated with unfavourable outcomes in pediatric AML patients. In previous studies, we found WASP-family verprolin-homologous protein-1 (WAVE1) was involved in the MDR mechanisms in K562/A02 leukemia cells. To investigate the expression of WAVE1, P-gp, MRP1, LRP/MVP and BCRP; and if WAVE1 is involved in MDR of human leukemia cell.

METHODSWAVE1, P-gp, MRP1, LRP, BCRP mRNA and protein expression in bone marrow mononuclear cells (BMMCs) were measured by real-time fluorescence quantitative PCR (RQ-PCR) and Western blot in a cohort of 52 children with acute myeloblastic leukemia. During follow-up, of the 52 patients, 21 were documented as being relapsing or refractory, and 31 were induced into complete continuous remission. Furthermore, HL60 cells and HL60/ADR cells were transiently transfected with PCDNA3.1-WAVE1 reconstructed plasmid and specifically siRNA to WAVE1 respectively, and the expression of WAVE1, MRP1 and BCRP before and after transfection was assessed by real-time PCR and Western blot analysis.

RESULTS(1) The expression levels of WAVE1, P-gp, MRP, LRP and BCRP in refractory/relapsing group were much higher than that in complete continuous remission (CCR) group. (2) WAVE1 mRNA and protein expression in BMMCs of children were at higher levels when they were newly diagnosed or relapsed, compared with complete continuous remission. (3) The WAVE1 expression at mRNA and protein level in HL60/ADR cells was increased by about 353% and 95% respectively as compared with that in HL60 cells. (4) Overexpression of WAVE1 in HL60 cell lines upregulated the expression levels of MRP and BCRP (MRP mRNA and protein level were increased by about 16.54 times and 129% respectively, BCRP was increased by 4.93 times and 96%); whereas suppression of WAVE1 expression by RNA interference downregulated the expression levels of MRP1 and BCRP (MRP mRNA and protein level was only 11% and 43% of pre-disturbance respectively, BCRP was 14% and 71%).

CONCLUSIONSHigher levels of WAVE1 in the BM indicate an unfavorable prognosis in children with AML. WAVE1 is related to the development of AML and involved in the MDR mechanisms, and regulates the level of MRP1 and BCRP.

Adolescent ; Child ; Child, Preschool ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Infant ; Leukemia, Myeloid, Acute ; genetics ; Male ; RNA, Small Interfering ; Wiskott-Aldrich Syndrome Protein Family ; genetics