OBJECTIVETo investigate the inhibitive effects of small interfering RNA (siRNA) on hepatitis C virus (HCV) replication in cells infected by HCV in vitro.

METHODSThe HCV RNA transcripts prepared by pFL-JC1 were transfected into Huh-7.5.1 cells. Na ve Huh-7.5.1 cells were incubated with the supernatants of transfected cells and the expression of HCV core protein in infected cells was detected by indirect immunofluorescence. The infected cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA for 24 h, 48 h and 72 h, respectively. The normal Huh-7.5.1 cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA. Group of blank, lipofectamine 2000, unrelated siRNA and IFNα-2b (1000 IU/ml) served as controls. The HCV RNA and PKR mRNA levels were examined by quantitative RT-PCR.

RESULTSThe HCV core protein in HCV infected cells was detected. Compared with control groups, the HCV RNA levels in infected cells significantly decreased when transfected with 40 and 200 nmol/L of siRNA for 24 h; 4, 40 and 200 nmol/L of siRNA for 48 h and 72 h (P<0.05). The HCV RNA levels in infected cells treated with IFNα-2b (1000 IU/ml) for 24 h, 48 h and 72 h were significantly lower than those in control groups (P<0.05 or P<0.01). The PKR mRNA levels in Huh-7.5.1 cells transfected with siRNA of three concentrations did not have significant difference, as compared with control groups (P>0.05).

CONCLUSIONsiRNA against HCV NS5B region can effectively inhibit HCV replication in HCV infected cells, but can not activate the dsRNA-dependent protein kinase (PKR).

Cell Line, Tumor ; Hepacivirus ; drug effects ; genetics ; physiology ; Humans ; RNA, Small Interfering ; pharmacology ; Transfection ; Viral Nonstructural Proteins ; genetics ; Virus Replication ; drug effects

Hydrophilic low molecular drugs, peptides and proteins, which are always poor in bioavailability, are mainly absorbed through the paracellular way in which the tight junction is the elementary framework. The tight junctions are a multiple unit structure composed of multiprotein complex that affiliates with the underlying apical actomyosin ring. Tight junction proteins are identified including transmembrane proteins (occludin, claudin and JAM) , cytoplasmic plaque proteins (ZO-1, ZO-2, ZO-3 and cingulin) and cytoskeleton. Traditional absorption enhancers can usually impair mucous membranes which constraint the utilization of these enhancers. Recently, with the increasing knowledge of the structure and function of tight junctions, many new absorption enhancers have been developed such as NO donor, CPE, Zot, and so on. In vivo and in vitro studies have shown that these enhancers could be effectively used to increase the absorption of paracellular markers and low bioavailable drug across intestinal epithelium with lower side effect. In short, the transient opening of the tight junctions by these enhancers provides new ideas that could help in novel drug delivery of therapeutic agents.
Animals ; Biological Availability ; Cell Adhesion Molecules ; metabolism ; Cholera Toxin ; pharmacology ; Claudin-1 ; Cytoskeleton ; metabolism ; Decanoic Acids ; pharmacology ; Drug Delivery Systems ; Enterotoxins ; pharmacology ; Humans ; Intestinal Absorption ; drug effects ; Membrane Proteins ; metabolism ; Nitric Oxide Donors ; pharmacology ; Occludin ; Phosphoproteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Tight Junctions ; metabolism ; physiology ; Zonula Occludens-1 Protein

OBJECTIVETo study the inhibition of the expression of bcl-xL gene induced by RNA interference in CNE-2Z cell line in addition to the inhibition of its proliferation and apoptotic induction.

METHODSSmall interfering RNAs targeting bcl-xL gene were synthesized by using web design software provided by Amnion and the silencer short interfering RNA (siRNA) construction kit; fluorescein-labeled siRNAs were done by FAM-silencer siRNA labeling kit; siRNAs were transfected into CNE-2Z cells by using lipofectamine 2000 reagent; siRNA transfection efficiencies were analyzed by fluorescent microscopy; down-regulation of bcl-xL was detected by RT-PCR; thiazolyl blue (MTT) assay was used to assess the cell growth; apoptosis of CNE-2Z cells was analyzed by flow cytometry.

RESULTSGreen fluorescence in the cells was seen clearly in FAM-labeled siRNA transfected group under the fluorescent microscope while none in the untransfected group. Different down-regulations of bcl-xL mRNA expression were found in the transfected groups. The expression of bcl-xL mRNA decreased by 10% - 70% in the siRNAs transfected CNE-2Z by RT-PCR scan analysis. The inhibitory rate of cell proliferation depended on time and concentrations to some extent. Different cell apoptosis could be induced by different concentrations of siRNA4.

CONCLUSIONSThe synthesized siRNAs in vitro were able to down-regulate the expression of bcl-xL There were different capabilities of the specific siRNAs down-regulation. The transient transfected bcl-xL siRNA4 could effectively inhibit the growth of the cancer cells and induce theirs apoptosis. It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for anti-nasopharyngeal carcinoma gene therapy.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Messenger ; genetics ; Transfection ; bcl-X Protein ; genetics

BACKGROUNDAssociations between glutamine (Gln) enriched nutrition support and surgical patients with gastrointestinal (GI) tumor remain controversy. The purpose of this meta-analysis was to assess the effect of Gln enriched nutrition support on surgical patients with GI tumor in term of relevant biochemical indices, immune indices, and clinical outcomes.

METHODSSix databases were systematically searched to find eligible randomized controlled trials (RCTs) from 1966 to May 2014. When estimated the analysis indexes, the relative risk (RR) was used as the effect size of the categorical variable, while the weighted mean difference (MD) was used as the effect size of a continuous variable. Meta-analysis was conducted with Rev Man 5.2.

RESULTSThirteen RCTs, involving 1034 patients, were included in the meta-analysis. The analysis showed that Gln enriched nutrition support was more effective in increasing serum albumin (MD: 0.10; 95% confidence interval [CI]: 0.02-0.18; P < 0.05), serum prealbumin (MD: 1.98; 95% CI: 1.40-2.55; P < 0.05) and serum transferring (MD: 0.35; 95% CI: 0.12-0.57; P < 0.05), concentration of IgG (MD: 1.26; 95% CI: 0.90-1.63; P < 0.05), IgM (MD: 0.18; 95% CI: 0.11-0.25; P < 0.05), IgA (MD: 0.22; 95% CI: 0.10-0.33; P < 0.05), CD3 + (MD: 3.71; 95% CI: 2.57-4.85; P < 0.05) and CD4/CD8 ratio (MD: 0.27; 95% CI: 0.12-0.42; P < 0.05). Meanwhile, it was more significant in decreasing the incidence of infectious complications (RR: 0.67; 95% CI: 0.50-0.90; P < 0.05) and shortening the length of hospital stay (MD: -1.72; 95% CI: -3.31--0.13; P < 0.05).

CONCLUSIONSGlutamine enriched nutrition support was superior in improving immune function, reducing the incidence of infectious complications and shortening the length of hospital stay, playing an important role in the rehabilitation of surgical GI cancer patients.

Enteral Nutrition ; Gastrointestinal Neoplasms ; surgery ; Glutamine ; therapeutic use ; Humans ; Parenteral Nutrition ; Postoperative Complications ; prevention & control ; Randomized Controlled Trials as Topic

Objective: This study aimed to identify the risk factors for genitourinary fistulas and delayed fistula recognition after radical hysterectomy for cervical cancer. Methods: This study was a retrospective analysis of data collected in the Major Surgical complications of Cervical Cancer in China (MSCCCC) database from 2004–2016. Data on sociodemographic characteristics, clinical characteristics, and hospital characteristics were extracted. Differences in the odds of genitourinary fistula development were investigated with multivariate logistic regression analyses, and differences in the time to recognition of genitourinary fistula were assessed by Kruskal–Wallis test. Results: In this study, 23,404 patients met the inclusion criteria. Surgery in a cancer center, a women’s and children’s hospital, a facility in a first-tier city, or southwest region, stage IIA, type C1 hysterectomy, laparoscopic surgery and ureteral injury were associated with a higher risk of ureterovaginal fistula (UVF) (p<0.050). Surgery in southwest region, bladder injury and laparoscopic surgery were associated with greater odds of vesicovaginal fistula (VVF) (p<0.050). Surgery at cancer centers and high-volume hospitals was associated with an increase in the median time to UVF recognition (p=0.016; p=0.005). International Federation of Gynecology and Obstetrics (FIGO) stage IIA1-IIB was associated with delayed recognition of VVF (p=0.040). Conclusion Intraoperative urinary tract injury and surgical approach were associated with differences in the development of UVFs and VVFs. Patients who underwent surgery in cancer centers and high-volume hospitals were more likely to experience delayed recognition of UVF. Patients with FIGO stage IIA1-IIB disease were more likely to experience delayed recognition of VVF.

It has been a long time since ultrasound hyperthermia began to be used in the clinical management of cancers and benign diseases. Numerous biological and clinical investigations have demonstrated that: hyperthermia in the range of 41-45 degrees C can significantly enhance clinical response to radiation therapy and chemotherapy, and high-temperature hyperthermia (greater than 65 degrees C) alone is now being used as an alternative to conventional invasive surgery for selective tissue destruction, causing tumor coagulation and necrosis. As a promising noninvasive and effective local therapy, HIFU has attracted great attention. China is advanced in the clinical applications of HIFU. This article gives an introduction of the development and applications of ultrasound hyperthermia technology, and also provides a general review of a selection of ultrasound hyperthermia systems both in clinical use and under development.
Equipment Design ; Humans ; Hyperthermia, Induced ; instrumentation ; methods ; Neoplasms ; therapy ; Ultrasonics ; Ultrasound, High-Intensity Focused, Transrectal

OBJECTIVETo systematically assess the effect of early enteral nutrition support after gastrointestinal operation on prognosis.

METHODSThe Cochrane Library, PubMed, CBM, CNKI, Wanfang, and VIP databases were retrieved via computer system for randomized controlled trails(RCTs) with early enteral nutrition support to patients undergoing gastrointestinal operation. Quality of studies was evaluated by the Cochrane Jadad rating scale. Nutrition indexes, bowel function indices, postoperative complications, health-economics indices were collected. Meta-analysis was conducted with RevMan 5.2.

RESULTSEleven relevant RCTs studies with 1087 cases were enrolled, including 541 patients in the study group(early enteral nutrition) and 546 in the control group. Meta-analysis showed that patients in the study group had significantly higher levels of plasma albumin and prealbumin than those in the control group(WMD=2.87, 95%CI:1.03-4.71; WMD=0.04, 95%CI:0.02-0.05). The time of postoperative bowel ventilation in the study group was significantly shorter than that in the control group(WMD=4.10, 95%CI:-5.38--2.82). The postoperative complication rate in the study group was significantly lower as compared to the control group(RR=0.64, 95%CI:0.44-0.93).

CONCLUSIONEarly enteral nutrition support after gastrointestinal operation is safe and effective, which can improve the nutritional status, promote bowel function return, and reduce postoperative complication rate.

Digestive System Surgical Procedures ; Enteral Nutrition ; Gastrointestinal Diseases ; surgery ; Humans ; Postoperative Complications ; Prognosis ; Randomized Controlled Trials as Topic

Cystic tumour of the atrioventricular node is a rare primary cardiac tumour that can cause complete heart block and sudden death. Here, we describe a male case aged 42 years who suddenly died without a medical and family history of cardiac illnesses. After detailed macroscopic and microscopic examinations, a cystic mass was found in the atrioventricular nodal region. The small lesion was less than 1 cm in diameter, and consisted of small and large cystic spaces and tubular structures lined by flat, cuboidal or squamous epithelium. Immunohistochemical staining revealed the tumour epithelium positive for epithelial membrane antigen, carcinoembryonic antigen, antigen epitopes AE1/AE3, cytokeratins CK5/6 and CK7, but negative for calretinin, HBME-1, Wilms' tumor 1, factor VIII, chromogranin, synaptophysin or smooth muscle actin, suggesting an endodermal rather than mesothelial origin.
Adult ; Atrioventricular Node ; metabolism ; pathology ; Heart Neoplasms ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Male

BACKGROUND/AIMS: The co-occurrence of obesity aggravates asthma symptoms. Diet-induced obesity increases helper T cell (TH) 17 cell differentiation in adipose tissue and the spleen. The 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor pravastatin can potentially be used to treat asthma in obese patients by inhibiting interleukin 17 (IL-17) expression. This study investigated the combined effects of pravastatin and anti-IL-17 antibody treatment on allergic inflammation in a mouse model of obesity-related asthma. METHODS: High-fat diet (HFD)-induced obesity was induced in C57BL/6 mice with or without ovalbumin (OVA) sensitization and challenge. Mice were administered the anti-IL-17 antibody, pravastatin, or both, and pathophysiological and immunological responses were analyzed. RESULTS: HFD exacerbated allergic airway inflammation in the bronchoalveolar lavage fluid of HFD-OVA mice as compared to OVA mice. Blockading of the IL-17 in the HFD-OVA mice decreased airway hyper-responsiveness (AHR) and airway inflammation compared to the HFD-OVA mice. Moreover, the administration of the anti-IL-17 antibody decreased the leptin/adiponectin ratio in the HFD-OVA but not the OVA mice. Co-administration of pravastatin and anti-IL-17 inhibited airway inflammation and AHR, decreased goblet cell numbers, and increased adipokine levels in obese asthmatic mice. CONCLUSIONS: These results suggest that the IL-17–leptin/adiponectin axis plays a key role in airway inflammation in obesity-related asthma. Our findings suggest a potential new treatment for IL-17 as a target that may benefit obesity-related asthma patients who respond poorly to typical asthma medications.
Adipokines ; Adipose Tissue ; Animals ; Asthma* ; Bronchoalveolar Lavage Fluid ; Cell Differentiation ; Diet, High-Fat ; Goblet Cells ; Humans ; Inflammation* ; Interleukin-17 ; Mice ; Obesity ; Ovalbumin ; Ovum ; Oxidoreductases ; Pravastatin ; Respiratory Hypersensitivity ; Spleen

OBJECTIVETo screen the differentially expressed proteins especially at the precancerous stage of diethylnitrosamine (DEN) induced hepatocarcinogenesis by comparative proteome research.

METHODSRats were divided into normal and DEN groups and sacrificed periodically. The liver samples were stained with gamma-glutamyl transpeptidase (GGT) and HE to distinguish the preneoplastic lesion (pre-HCC) from the normal and HCC tissues. The two-dimensional electrophoresis (2-DE) and mass spectrometry (MALDI-TOF-MS/MS) were then applied to analyze the differentially expressed protein between pre-HCC and normal tissues, pre-HCC and HCC, as well as HCC and normal tissues. A few of the candidate proteins such as laminin receptor 1 (67LR) and agmatinase were validated by Western blot and RT-PCR.

RESULTSTotally, there were 82 proteins that differentially expressed two fold or more in one kind of tissues sample than the other, 47 of which occurred in the pre-HCC tissues. Eight proteins including 67LR were consistently up-regulated from normal tissue to pre-HCC and then to HCC tissues, while 22 proteins including agmatinase showed progressively down-regulated in these tissues samples.

CONCLUSIONThe protein expression profiles are different during the process of hepatocarcinogenesis. Further study on the differentially expressed protein, especially these upregulated in the precancerous stage such as 67LR and agmatinase, might contribute to prevention and early diagnosis of human HCC.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; chemically induced ; metabolism ; pathology ; Diethylnitrosamine ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; chemically induced ; metabolism ; pathology ; Male ; Neoplasm Proteins ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; Proteins ; metabolism ; Proteome ; Rats ; Rats, Wistar ; Receptors, Laminin ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Ureohydrolases ; metabolism ; gamma-Glutamyltransferase