Objective To investigate the relationship between refractory hypertension and renal hemodynamics in end stage renal diseases (ESRD) patients.Methods ESRD patients were classified into:patients with refractory hypertension (group A) and patients with normal blood pressure(group B).Renal hemodynamic indices were ex- amined by duplex ultrasonography.Fasting serum lipid (TC,TG,HDL-C,LDL-C,Lp(a),ox-LDL) and serum parathyroid hormane (PTH) were determined in all patients.Results Significant differences were found in renal hemodynamic indices such as peak systolic velocity (PSV),mean flow velocity (MV),pulsatility index (PI),renal- aortic ratio (RAR) and in clinical index such as Lp(a) and ox-LDL between the two group.Refractory hyperten- sion patients had lower renal hemodynamic indices and higher Lp(a) and ox-LDL levels than in patients with con- trolled BP.Logistic regression analysis revealed that refractory hypertension was related with PSV,EDV,Pl and RAR,but not relevant with sex,age,dialysis time,hematocrit,BUN,creatinine,TC,TG,HDL-C,LDL-C, PTH,MV and RI.Conclusion Atherosclerotic renal artery stenosis and severe disorder in renal hemodynamics is likely the cause for refractory hypertention in ESRD patients.The rise of serum Lp(a) and ox-LDL might acceler- ate renal artery atherosclerosis.

The aim of this study was to identify some biomarkers for predicting lymph node metastasis and prognosis of human epidermal growth factor receptor 2 (Her-2)-positive breast cancer (BC). We analyzed correlations between microRNAs (miRNAs) and the prognosis of patients with BC based on data collected from The Cancer Genome Atlas (TCGA) database. The expression levels of miR-455, miR-143, and miR-99a were measured in clinical samples of Her-2-positive BC patients with different degrees of lymph node metastasis. We investigated the impacts of overexpressed miR-455 on the proliferation and invasiveness of MDA-MB-453 cells and measured its effects on the expression of long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of miR-455 was significantly and positively correlated to the prognosis and overall survival (OS) of the BC (P=0.028), according to TCGA information. The expression level of miR-455 was positively correlated with OS and relapse-free survival (RFS) of patients with Her-2- positive BC, and was negatively correlated with the number of metastatic lymph nodes (P 0.05). Transwell assay suggested that MDA-MB-453 cells became much less invasive (P 0.01) after being transfected with miR-455 mimics. During the qRT-PCR, the expression level of MALAT1 declined significantly after transfection (P 0.01). Overexpressed miR-455 significantly inhibited the proliferation and migration of MDA-MB-453 cells and the expression of MALAT1. We conclude that miR-455 may be a useful potential biomarker for forecasting lymph node metastasis and the prognosis of Her-2-positive BC patients. miR-455 may play an important role in lymph node metastasis of BC by interacting with MALAT1.

OBJECTIVETo provide a basis for development and preparation of the new anti-tumor agents from Cortex A-canthopanacis senticosus (CAS), through isolating the active substances from CAS and studying the anti-tumor effect of CAS extracts in vivo and in vitro.

METHODSThe effects of CAS extracts and its isolated ingredients on tumor cell proliferation in vitro was determined by 3H-TdR incorporation; the anti-tumor component of CAS was isolated and purified by chromatography; the tumor bearing mice model was established by injecting tumor cell subcutaneously, and the model was used to observe the anti-tumor effect of CAS extract administered through gastrogavage.

RESULTSCAS extract showed obvious inhibition on tumor cell proliferation originated from multiple tissues (P < 0.01) and displayed a better dose-effect relationship. After orally taken CAS extract, the general condition of mice in the experimental group were better than that in the untreated control group, revealing a slower growth and significantly prolonged survival period (P < 0.01). A protein component, having inhibitory effect on tumor cell proliferation and the molecular weight was 64 kda, it was isolated by the thin layer gel chromatography.

CONCLUSIONCAS has not only the in vitro inhibitory effect on cell proliferation of multiple kinds of tumor, but also a good anti-tumor effect in vivo. The anti-tumor activity of CAS is correlated with a protein component with the molecular weight of 64 kda. Further isolation, purification, study on mechanism will provide scientific evidence for clinical application and development of CAS in anti-tumor effect.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; Female ; HL-60 Cells ; pathology ; Humans ; Leukemia, T-Cell ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Tumor Cells, Cultured

OBJECTIVETo study the effect of Astragalus membranaceus extract (AME) in regulating the immune function of human peripheral blood immune cells (PBIC) in vitro.

METHODSEffects of AME on the proliferation activity of peripheral blood mononuclear cells (PBMC) and the tumor cell phagocytosis of peripheral blood adherent monocytes (PBAM) were measured by using 3H-TdR incorporation. Effect of the tumor-killing activity of cytotoxic T-lymphocyte (CTL) was determined by using 51Cr-releasing assay. Effects on production of IgG by peripheral blood B cells (PBBC) and IL-6 by PBAM were tested by means of ELISA, and effect on production of TNF-alpha by PBAM was studied by means of biological method. Besides, the protein elements of AME were analysed by SDS-PAGE.

RESULTSAME could promote the proliferation of human PBMC, elevate the tumor cell-killing activity of CTL, strengthen the tumor cell phagocytosis and cytokines (TNF-alpha and IL-6) production of PBAM, and promote the IgG production of PBBC. AME contained multiple protein elements.

CONCLUSIONAME has effect in enhancing human immuno-function and anti-tumor activity, it could be applied in clinical practice for immuno-modulation and tumor treatment.

Adjuvants, Immunologic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Astragalus membranaceus ; chemistry ; Cytotoxicity, Immunologic ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; In Vitro Techniques ; Interleukin-6 ; blood ; Leukocytes, Mononuclear ; immunology ; Phagocytosis ; drug effects ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo identify enterovirus 71 (EV71) strains isolated from patients with hand, foot and mouth disease (HFMD) in Guangdong and Fujian provinces from 2000 to 2001 by using phylogenetic analysis.

METHODSAll 25 samples were first tested for enteric viruses by RT-PCR using enterovirus specific primers EV-1 and EV-2, and then were identified for EV71 by RT-PCR using EV71 specific primers 159S and 162A. The amplicons of 485bp segment (part of the VP1 gene) were cloned into pGEM-T and sequenced. A phylogenetic tree was constructed by comparison of the sequences with other 12 EV71 strains isolated from China, Japan, Hungary, and the United States including the prototype BrCr.

RESULTSThe positive rate of EV71 was about 20%. The sequence analysis showed that the new isolate (GZH2000) shared 94%-96% nucleotide identity with three strains isolated in 1998 and 2000, and 91% with a strain isolated in 1987 from Chinese mainland, but shared only 82%-84% homology with EV71 isolates studied abroad.

CONCLUSIONSEV71 is one of the important pathogens of HFMD in south China. The strains isolated from mainland were closely related with most isolates from Taiwan, but different from most EV71 strains reported abroad. The symptoms of EV71 infection in mainland were not as intensive as those described in Taiwan's outbreak.

Adolescent ; Child ; Child, Preschool ; China ; Enterovirus ; genetics ; isolation & purification ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Phylogeny ; Sequence Analysis ; Sequence Homology, Nucleic Acid

It was expected that recombinant Aspergillus niger glucose oxidase could be expressed in Trichoderma reesei with stable activity. T. reesei CBHI promoter--CBHI ss. gene--A. niger glucose oxidase gene--T. reesei CBHI terminator--A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator--pUC19 (pCBHGOD) vector was constructed in E. coli DH5alpha by PCR application and gene cloning methods. T. reesei QM9414 protoplast was transformed by T. reesei CBHI promoter-CBHI ss. Gene--A. niger glucose oxidase gene--T. reesei CBHI terminator-A. nidulans gpd promoter--E. coli Hygromycin B phosphotransferase gene--A. nidulans trpC terminator linear DNA fragment (CBHGOD fragment) that was made by digestion of pCBHGOD with Kpn I. T. reesei mutant clone with homologous recombinant A. niger glucose oxidase gene was selected by PCR method. Recombinant glucose oxidase was produced by mutant T. reesei strain under induction of wheat straw for 5 days. Recombinant glucose oxidase molecular mass was showed the same as native A. niger glucose oxidase standard from Sigma company by Western blot analysis. Recombinant glucose oxidase activity was 25u/mL in medium. The yield was 0.5 g/L in comparison with Sigma company glucose oxidase standard. There was no recombinant GOD degradation during Trichoderma reesei cultivation that was showed in Western blot analysis. Trichoderma reesei has capability to be a new recombinant host for Aspergillus niger GOD production.
Aspergillus niger ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Glucose Oxidase ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Trichoderma ; genetics ; metabolism

BACKGROUNDFor patients with severe endometriosis, the spontaneous pregnancy rates have been reported to be near 0 due to extreme distortion of normal pelvic anatomy. Surgery is one of the treatment options; however, if patients failed to conceive after surgery, in vitro fertilization (IVF) is effective. The objective of this retrospective study was to determine the clinical characteristics of IVF/intracytoplasmic sperm injection (ICSI) in patients with stage III/IV endometriosis, and to determine the impact of the interval from surgery to IVF/ICSI on outcome.

METHODSOne hundred and sixty patients who were diagnosed with stage III/IV endometriosis underwent IVF/ICSI cycles between February 2004 and June 2009 were enrolled. The mean interval from surgery to IVF, number of oocytes retrieved, fertilization rate, implantation rate, embryos transferred, and good embryos transferred were compared between two age groups (35 years).

RESULTSThe mean interval from surgery to IVF was (37.9+/-28.9) months for the group35 years of age. Twenty-five IVF/ICSI cycles (12.8%) were performed during the first year after surgery, and 34.9% IVF/ICSI cycles were performed 2 years after surgery. No significant differences existed between the two groups with respect to the fertilization rate, implantation rate, number of embryos transferred, number of good embryos, clinical pregnancy rates, live birth rates, and cumulative clinical pregnancy rates (P>0.05). The probability of cumulative clinical pregnancies was 75%, 50%, and 25% ((29.0+/-4.8), (61.0+/-7.6), and (120.0+/-16.9) months after surgery, respectively).

CONCLUSIONSFor infertile patients with stage III/IV endometriosis, the optimal time to conceive by IVF/ICSI is <2 years after surgery; nevertheless, most of the patients took a longer time to conceive.

Adult ; Endometriosis ; surgery ; Female ; Humans ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Time Factors

Objective To investigate the impacts of LINC00943 on proliferation,apoptosis,and invasion of esophageal squamous cell carcinoma(ESCC)cells by regulating the miR-126-5p/HOXB2 axis.Methods QRT-PCR was applied to detect the expression of LINC00943 in 45 ESCC tissues and cell lines;The effects of LINC00943 knockdown on the proliferation,apoptosis and invasion of KYSE30 cells were detected by MTT assay,flow cytometry and Transwell chamber.Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti apoptotic factor B cell lymphomatoma-2(Bcl-2),cleaved caspase-3,matrix metalloproteinase-2(MMP-2),MMP-9,and HOXB2;dual luciferase reporter gene experiment was applied to verify the relationship between miR-126-5p and LINC00943 and HOXB2;the ESCC nude mouse in vivo model was constructed and separated into si-NC and si-LINC00943 groups,the tumor mass and volume were measured,qRT-PCR was applied to detect the expression of miR-126-5p in transplanted tumor tissue,immunohistochemistry was applied to detect the expression of HOXB2 protein in transplanted tumor tissue,and TUNEL staining was applied to detect cell apoptosis in tumor tissue.Results The expression of LINC00943 mRNA increased in ESCC tissues and cell lines(P＜0.05);Compared with the si-NC group,the proliferation activity,migration and invasion cell number,HOXB2,PCNA,MMP-2,MMP-9 and Bcl-2 protein expression of KYSE30 cells in the si-LINC00943 group were decreased,and the expression of miR-126-5 p,Bax,Cleaved Caspase-3 and apoptosis rate were increased(P＜0.05);downregulation of miR-126-5p was able to weaken the inhibitory effect of LINC00943 knockdown on the malignant behavior of KYSE30 cells(P＜0.05);dual luciferase reporter gene experiment confirmed the targeted regulatory relationship between miR-126-5p and LINC00943,and miR-126-5p and HOXB2(P＜0.05);The ESCC nude mouse model was constructed in vivo and divided into si-NC and si-LINC00943 groups.The tumor mass,tumor volume,expression of miR-126-5p and HOXB2 in transplanted tumor tissues and apoptosis rate of tumor cells were measured.(P＜0.05).Conclusion LINC00943 is highly expressed in ESCC tissues and cell lines.Knocking down LINC00943 may inhibit the expression of HOXB2 by up-regulating the expression of miR-126-5p,promote the apoptosis of ESCC cells,and inhibit their proliferation,migration and invasion.

Objective To explore the relation between hepatitis B virus DNA load and genotype with the level of large envelope protein. Methods Serum HBV DNA was quantitively detected by using real time polymerase chain reaction (RT-PCR). The LHBs were detected by using enzyme linked immuno sorbent assay (ELISA) and HBV markers were detected by time differentiate immunofiuorescence assay in 140 serum samples collected from chronic hepatitis B patients. The genotypes of HBV were identified by DNA sequencing; and analyze their relationship. Results There was no significant difference between positive rate of LHBs and that of HBV DNA in HBeAg negative and positive group(P＞0.05); The HBV LHBs absorbency was markedly correlated with the HBV DNA load(R2=0.9267). The difference of HBV LHBs absorbency between HBV genotype B and C was not significant. Conclusions The close correlation between HBV LHBs absorbence and HBV DNA load illustrated that he level of serum LHBs can be used to estimate the state of HBV replication; and there is no relationship between HBV LHBs absorbency and genotypes. So HBV LHBs may be used as a new serological marker to detect HBV replication.

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKα/β and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKα/β level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKα/β. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.
Apoptosis* ; Aspirin ; Cell Survival ; Chickens* ; Heat Stress Disorders ; Heat-Shock Proteins* ; Hot Temperature* ; HSP90 Heat-Shock Proteins ; In Vitro Techniques* ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Transducers