Objective The use of in vivo cryotechnique (IVCT) in combination with electrocardiograph (ECG) to study cardiac microcirculation under different hemodynamic conditions in living mouse.Methods Living mouse heart monitored by electrocardiograph was suffered from IVCT and freezing substitution under normal blood flow,myocardial ischemia or cardiac arrest conditions.Hematoxylin eosin (HE)staining,Schiff's staining and immunofluorescence staining for serum albumin,immunoglobulin were utilized on continuous paraffin sections,respectively.Confocal microscopy and statistical analyses were used.Results Comparing with normal hemodynamics,microvascular red cell volume reduction,morphology changed,myocardial cell glycogen loss,serum albumin ectopic distribution to myocardial cytoplasm,T tubular network failure and spacing width were happened in myocardial iscbemia condition; different shapes of red blood cells,myocardial cells glycogen deficiency,T tubular network failure and interval narrowing were found under cardiac arrest conditions.Conclusions Cardiac microcirculation,pathological changes of myocyte and its surrounding microenviroument in living mouse heart can be immediately captured in situ by the application of IVCT and ECG.

Objective Vaccination is a most effective method for the prevention of severe diseases caused by pandemic influenza and microRNA ( miRNA) mediated gene silencing has offered a novel approach to the construction of new vaccines.Our study aimed to construct a recombinant influenza A ( H1 N1 ) virus with the PB1 gene that carries the target fragment of miRNA Let-7b. Methods After comparing the sequence of the A/Nanjing/108/2009 H1N1 viral fragments with that of Let-7b, we selected PB1 as the optimal gene sequence, inserted the Let-7b binding target gene into PB1, ligated the modified fragments with pDP 2000, and named the recombinant plasmids pDP-mu-PB1 and pDP-sclb-PB1, respectively.We co-transfected the MDCK and 293T cells with the recombinant and other seven plasmids and injected the supernatant into the allantoic cavity of the chickenembryo for virus propagation, followed by detection of the virus by hemagglutination ( HA) assay and measurement of the viral titer by TCID50 .We amplified the viral cRNA by RT-PCR and identified the viruses by agarose gel electrophoresis and nucleotide sequence analysis. Results PB1 was the optimal sequence ( 83 bp -107bp) for the attenuation of viruses.The HA-titers of miRT-H1N1 and scbl-H1N1 were 1∶32 and 1∶64, and their viral loads were 4.68 ×105 and 7.94 ×104 TCID50/mL, respectively.Nucleotide sequence analysis showed the expected fragment in the rescued virus. Conclusion A recombinant strain vaccine was successfully constructed, which has laid the foundation for fur-ther assessment of virulence.

AIM: To evaluate the specific cellular immune response in mice inoculated with the recombinant hepatitis B virus (HBV) surface vaccine (rHBs). METHODS: Spleen T lymphocyte reactivity to rHBs was assessed by a proliferation assay and cytokine secretion. BALB/c mouse were intraperitoneally inoculated with rHBs at doses of 0.65, 1.25, 2.5 or 5 ?g for once or twice. 4 weeks later, spleen lymphocytes were harvested and restimulated with rHBs in experimental group or with PBS as control. The spleen lymphocytes were labeled with [~3H]-thymidine for 3 days. The [~3H]-thymidine incorporation was measured, which expressed as the incorporation of [~3H] (counts?min~ -1 ) and stimulation index (SI) was calculated by the method of dividing the cpm obtained in the experimental group by that in control group. The content of IL-2 and IFN-? secreted from the spleen lymphocyte were measured by the method of ELISA. RESULTS: 2 weeks after primary vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.55, 1.93, 2.41, 2.81 ng/L, respectively. IL-2 was 5.48?8.88, 9.28?6.98, 28.53?14.32, 64.69?20.88 ng/L, respectively. IFN-? was 8.22?8.61, 9.89?9.34, 20.27?15.50, 30.77?22.12 ng/L, respectively. 2 weeks after boost vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.61, 2.05, 3.74, 3.62 ng/L, respectively. The IL-2 was 5.75?5.04, 102.53?67.52, 177.13?91.12, 332.10?124.31 ng/L, respectively. IFN-? was 3.63?4.42, 28.33?13.04, 59.66?25.75, 80.73?19.30 ng/L, respectively. CONCLUSION: Specific proliferation activity and IL-2, IFN-? secretion from the spleen lymphocytes of the mouse inoculated with rHBs are produced,that the strength is dependent on the dose of vaccination.

AIM To study the chemical constituents from Vigna umbellata Ohwi et Ohashi.METHODS The ethyl acetate fraction of 80％ ethanol extract from V.umbellata was isolated and purified by silica,Sephadex LH-20 and ODS column,then the structures of obtained compounds were identified by spectral data and physicochemical properties.RESULTS Twelve compounds were isolated and elucidated as (+) catechin (1),(-) epicatechin (2),3-furanmethanol-β-D-glucopyranoside (3),myricetin-3-O-β-D-glucopyranoside (4),quercetin7-O-β-D-glucopyranoside (5),(+) catechin-3-O-β-D-glucopyranoside (6),(+) catechin-5-O-β-D-glucopyranoside (7),quercetin-3 '-O-α-L-rhamnoside (8),(±) dihydroquercetin (9),quercetin (10),ethyl gallate (11),propanediol (12).CONCLUSION All the compounds are isolated from V.umbellata for the first time,and compounds 4,8,9,11,12 are first obtained from genus Vigna.

Objective: To establish a method for determing the trichloroethylene(TCE)and trichloroethanol(TCOH)in blood samples by liquid-liquid extraction-gas chromatography with electron capture detector. Methods: With this method,ether was used as extraction solvent and trichloromethane was used as an internal standard. The whole blood sample was extracted with ether, and dehydrated by anhydrous sodium sulfate. Then the analytes were separated on HP-5 capillary column(30m×0.32mm×0.15μm)and detected byECD.The retention time was for qualitative analysis and the internal standard was for quantitation. Results: The standard curves of TCE and TCOH showed significant linearity between 95.5μg/L-7640.0μg/L(r=0.9997)and 19.0μg/L-1520.0μg/L(r=0.9992). The average recovery was 95.5%-103.6%.The intra-day and inter-day precisions(RSD)were 2.5%-6.8%(n=6)and 1.6%-4.3%(n=6) respectively. The detect limit of TCE and TCOH were 2.10 μg/L and 0.56μg/L(S/N=3)respectively.The blood can be kept 7 days at-20℃ refrigerator without significantly loss. Conclusion This method is proved to be simple,practical and highly sensitive. It can satisfy the request for the determination of blood samples of humans exposed to TCE.

Objective: To establish a method for determination the S-phenylmercapturic acid in urine by dispersive solid-phase extraction using Humic Acid/Fe₃O₄ magnetic nanocomposite as adsorbent. Methods: The 5 ml of urine samples were adjusted to pH 1.0 and extracted by Fe3O4@HA. Then the analytes were separated on EC-C₁₈ capillary column and detected by HPLC-VWD. The S-phenylmercapturic acid was characterized by the retention time and quantified by peak area and external standard method. Results: The standard curves of SPMA showed significant linearity between 0.04~1.00 mg/L (r=0.999 7) . The average recovery was 94.2%~102.4%. The intra-day and inter-day precisions (RSD) were 2.9~6.7% (n=6) and 3.1~7.5% (n=6) respectively. The detect limit of SPMA was 0.012 g/L (S/N=3) . Conclusion This method is simple, rapid, sensitive and accurate. It is applicable for determination of SPMA in the urine of works who were exposed to benzene.

PURPOSE: Effective biomarkers and models are needed to improve the prognostic prospects of clear cell renal cell carcinoma (ccRCC). The purpose of this work was to identify DNA methylation biomarkers and to evaluate the utility of DNA methylation analysis for ccRCC prognosis. MATERIALS AND METHODS: An overview of genome-wide methylation of ccRCC tissues derived from The Cancer Genome Atlas (TCGA) database was download for analysis. DNA methylation signatures were identified using Cox regression methods. The potential clinical significance of methylation biomarkers acting as a novel prognostic markers was analyzed using the Kaplan-Meier method and receiver operating characteristic (ROC) curves. RESULTS: This study analyzed data for 215 patients with information on 23171 DNA methylation sites and identified a two-DNA methylation signature (cg18034859, cg24199834) with the help of a step-wise multivariable Cox regression model. The area under the curve of ROCs for the two-DNA methylation signature was 0.819. The study samples were stratified into low- and high-risk classifications based on an optimal threshold, and the two groups showed markedly different survival rates. Moreover, the two-DNA methylation marker was suitable for patients of varying ages, sex, stages (I and IV), and histologic grade (G2). CONCLUSION: The two-DNA methylation signature was deemed to be a potential novel prognostic biomarker of use in increasing the accuracy of predicting overall survival of ccRCC patients.
Biomarkers ; Carcinoma, Renal Cell ; Classification ; DNA Methylation ; Genome ; Humans ; Methods ; Methylation ; Prognosis ; ROC Curve ; Survival Rate

How to give full play to the advantages of rehabilitation institutions, and to improve service quality and administration efficiency are some of the complex issues faced by the high-level administrators of rehabilitation institutions. This paper studied a total of 67 institutions to investigate the operation and management model of rehabilitation institutions in China, as well as their advantages and disadvantages.

The establishment of an efficient performance management system is crucial for a rehabilitation institution. This article discussed the concept of performance, performance management and ways to establish an efficient performance management system.

BACKGROUND: Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer. METHODS: Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38. RESULTS: lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer. CONCLUSION Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.
Humans ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic/genetics* ; MAP Kinase Signaling System ; MicroRNAs/metabolism* ; p38 Mitogen-Activated Protein Kinases/metabolism* ; RNA, Long Noncoding/metabolism* ; Triple Negative Breast Neoplasms/pathology*