AIM:To explore the effects of small interfering RNA(siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells growth and cycle.METHODS:Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent.The PCNA mRNA and PCNA protein were detected by real-time polymerase chain reaction(RT-PCR) and immunohistochemical method.Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA-siRNA transfected.Flow cytometry was used to observe the cell cycle.RESULTS:In CNE2 cells after PCNA-siRNA transfection,the expressions of PCNA mRNA and protein were down-regulated at different degree.Inhibition ratio of PCNA mRNA was 98.5%.Meanwhile,the cell cycle was suffocated at G0/G1 stage.CONCLUSION:The synthesized PCNA-siRNA effectively interferes nasopharyngeal carcinoma cells by down-regulating the expressions of the PCNA mRNA and its protein,therefore inhibits the growth of CNE2 cells.Future application of PCNA-siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.

A strain of thermotolerant halophilic bacteria YJ0238 was isolated from the salt well at 47℃ in the Kangning countryside where was located at the side of Lancang River in the Tibet Autonomous Region. Some physiological and biochemical properties were characterized.16S rRNA gene sequence of YJ0238 was amplified by PCR,and its nucleotide sequence was determined. Based on it’s physiological and biochemical properties,homology and phylogenetic analysis of 16S rRNA gene sequence,strain YJ0238 was identified as a subspecies of the species Idiomarina zobellii. The GenBank accession number of the 16S rRNA gene sequence of strain YJ0238 is EF693953. Until now,there were few reports on the study of high-temperature and high-salt microbial in domestics. The results of this study will provide research material and information for further studies in this area.

Objective To investigate the correlation of P,IFN-γlevel of artificial insemination during pregnancy and pregnancy outcome.Methods 468 patients with infertility were done artificial insemination treatment cycles.The dydrogesterone were taken orally after surgery to luteal support.The blood P and IFN-γlevel of 54 patients with pregnant were observed.Results The correlation analysis of P,IFN-γ level of normal pregnancy group showed that the P level was negatively correlated with IFN-γlevel (r =-0.594,P ＜ 0.05).In the biochemical pregnancy group,the P level was decreased from the ninth days begin,and compared with the normal pregnancy group,the difference was significant(t =12.319,P ＜0.05).In the abortion group,the P level was decreased from the sixteenth days begin,and compared with the normal pregnancy group,the difference was significant (t =7.663,19.315,all P ＜0.05).Compared with the normal pregnancy group,the IFN-γ level of the biochemical pregnancy group and abortion group was significantly increased,the difference was significant (P ＜ 0.05).Conclusion The use of luteal support methods to raise the level of P in vivo has a positive effect on the prevention of biochemical pregnancy and spontaneous abortion,and monitoring the blood P and IFN-γlevel after artificial insemination can predict the prognosis of patients.

An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.

Objective To determine the viral etiology and its epidemic features of community-acquired pneumonia ( CAP) among pediatric inpatients in Guangzhou.Methods A total of 1 539 children with CAP admitted in Guangzhou Women and Children’ s Medical Center during June 2012 and June 2013 were enrolled in the study.Throat swab specimens were collected, and fluorescence quantitative polymerase chain reaction ( FQ-PCR) was performed to detect 11 respiratory pathogens.SPSS 17.0 was used for data processing, and χ2 test was performed to compare the infection rates among different groups.Results Among 1 539 patients, 550 cases (35.7%) were infected with at least one pathogen, and 101 (6.6%) were infected with two or more pathogens.The most popular viral etiologies were respiratory syncytial virus (RSV) (102, 6.6%), rhinovirus (RHV)(101, 6.6%), adenovirus (ADV) (78, 5.1%), influenza virus A (IVA) (78, 5.1%) and bocavirus (HBOV) (74, 4.8%).RSV infection often occurred in children with age≤3 years, while ADV or IVA infection often occurred in those with age >3 years.RSV infection rate peaked in winter and spring, IVA infection rate peaked in spring and summer, while ADV and HBOV infection rates peaked in summer.The rate of multiple infections in critically ill children (22/135, 16.3%) was significantly higher than that in other CAP patients (79/1 404,5.6%,χ2 =116.049, P<0.01).Conclusions Viral infection is common in pediatric inpatients with CAP, and RSV infection is the most popular.Multiple infections are more often to cause critical conditions.

Objectives To compare the dose distribution of the whole body γ-knife and three-dimensional conformal radiotherapy (3D-CRT) in the treatment of primary hepatic carcinoma to provide evidence for clinical treatment choice.Methods Totally 35 patients with primary hepatic carcinoma treated with whole body γ-knife from March 2014 to July 2015 underwent three-dimensional conformal radiotherapy planning,and dose volume histograms were used to evaluate the dose distribution of target volumes and normal tissue,and moreover conformity index (CI) and homogeneity index (HI) were analyzed.Results Between the two treatment plans,D98 and the mean dose Da of GTV were higher in whole body γ-knife plan than those in three-dimensional conformal radiotherapy plan.For PTV,the V95 of γ-knife and three-dimensional conformal radiotherapy were (95.05±0.02)％ and (90.00±0.03)％ (P=0.00) respectively.Target coverage of whole body γ-knife was better.For organ at risk,γ-knife showed better protection of normal tissue.Three-dimensional conformal radiotherapy showed better HI (GTV:0.11±0.15 vs 0.23±0.02,PTV:0.14±0.04 vs 0.21±0.03),while more normal liver tissue received excessive exposure.The whole body γ-knife increased dose of target volumes and decreased dose of normal tissue,while three-dimensional conformal radiotherapy showed better HI.Conclusion Compared with three-dimensional conformal radiotherapy,the whole body γ-knife shows advantage in dose distribution but HI of target volumes.For clinical using,appropriate treatment needs to be chosen according to the patients' actual situation.

In order to build a protein expression system in a cold-adapted bacterium Acinetobacter sp. DWC6, a promoter probe vector was constructed based on the plasmid pBR322. A fragment containing the promoter of the beta-lactamase gene (the ampicillin resistance gene) in pBR322 was eliminated and replaced by a fragment comprizing a kanamycin resistance gene amplified from pJRD215. DNA fragment harboring in the Acinetobacter species specific ori was also inserted into the plasmid pBR322 to construct a promoter probe vector named pBAP1, which could replicate both in E. coli and in Acinetobacter sp. DWC6. The promoter selection library was constructed by randomly inserting genomic DNA fragment of Acinetobacter sp. DWC6 at upstream of reported gene, and target promoters were screened from genomic library on ampicillin selection plates. The function of pBAP1 and isolated promoters were determined by detection of the ampicillin sensitivity and the expression level of beta-lactamase in the host cell.
Acinetobacter ; genetics ; Adaptation, Physiological ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Cold Temperature ; DNA, Bacterial ; genetics ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Models, Genetic ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; beta-Lactamases ; genetics ; metabolism

OBJECTIVETo investigate the effect of prostaglandin E2 (PGE(2)) on the proliferation of cultured hepatocellular carcinoma cells and explore which subtypes of EP prostanoid receptor mediate the action.

METHODSRT-PCR was used to determine COX-2 and EP receptor mRNA expression levels in human hepatocellular carcinoma cell line Hep3B and human normal hepatocyte line QSG7701. Cell counting kit-8 (CCK-8) assay was employed to investigate the effect of PGE(2), selective EP2 receptor agonist butaprost and EP3/EP4 receptor agonist PGE1 alcohol on the proliferation of the cells.

RESULTSCOX-2 mRNA was highly expressed in Hep3B cells but scarcely in QSG7701 cells. Hep3B cells expressed the mRNAs for all the EP receptor subtypes, but EP2 and EP4 receptors were much more strongly expressed than EP1 and EP3 receptors. PGE(2) significantly promoted Hep3B cell proliferation in a time- and dose-dependent manner, and 10 µmol/L PGE(2) increased the cell proliferation by 22.57% (P<0.001) after a 48-h incubation; treatment with 0.1, 1.0, and 10 µmol/L PGE(2) for 72 h resulted in significantly increased cell proliferation by 12.13% (P<0.01), 17.58% (P<0.01) and 33.07% (P<0.001), respectively. EP2 receptor agonist butaprost (20 µmol/L) increased Hep3B cell proliferation by 21.96% (P<0.001), but the EP3/EP4 receptor agonist PGE(1) alcohol (2-20 µmol/L) exhibited no significant mitogenic effect in Hep3B cells, and 200 µmol/L PGE(1) alcohol decreased the cell viability.

CONCLUSIONSelective activation of EP2 receptor promotes Hep3B cell proliferation, indicating the predominant role of EP2 receptor in mediating the mitogenic effect of PGE2.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; genetics ; metabolism ; Dinoprostone ; pharmacology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Receptors, Prostaglandin E, EP2 Subtype ; genetics ; metabolism

OBJECTIVETo evaluate the diagnostic value of liver fibrosis markers and ultrasonic examination for determining compensated liver cirrhosis in patients with chronic hepatitis B, and screen applicable non-invasive diagnostic marker for compensated liver cirrhosis.

METHODSSerum hyaluronic acid (HA), Type III procollagen (PCIII), laminin (LN) and Type IV collagen (CIV) were measured from 350 patients with chronic hepatitis B, who were also detected with liver biopsy and ultrasonography. To determine the cut-off value of every serum liver fibrosis marker for diagnosing compensated liver cirrhosis, data was analysed with clinical epidemiology methods. Then evaluated and compared all the markers.

RESULTS85 out of 350 patients were diagnosed as compensated liver cirrhosis by liver biopsy, and 81 had liver cirrhosis images by ultrasonic examination. HA achieved the biggest area under the ROC curve. The cut-off values with best sensitivity and accuracy of HA, PCIII, LN and CIV were 154.35 microg/L, 198.44 microg/L, 137.58 microg/L and 100.80 microg/L respectively. The related diagnostic sensitivities of HA, PCIII, LN and CIV were 82.4%, 63.5%, 57.3% and 70.6%, specificities were 79.3%, 54.0%, 56.8%, 68.3%, and accuracies were 80.0%, 56.3%, 56.9%, 68.9%, respectively. Parallel tests could increase the diagnostic sensitivity, but decreased specificity and accuracy accordingly. Compared with other non-invasive diagnostic methods, HA was the best marker (mu > or =1.814, P<0.05). The level of HA at 119.17 microg/L was suitable for determining compensated cirrhosis, with a 87.1% sensitivity, 67.6% specificity, 72.3% accuracy, 46.25% positive predictive value and 94.7% negative predictive value.

CONCLUSIONAmong the non-invasive serum diagnostic markers for liver fibrosis and ultrasonic examination for cirrhosis image, HA is the best marker for diagnosing compensated liver cirrhosis.

Adolescent ; Adult ; Biomarkers ; blood ; Child ; Collagen Type IV ; blood ; Female ; Hepatitis B, Chronic ; complications ; Humans ; Hyaluronic Acid ; blood ; Laminin ; blood ; Liver Cirrhosis ; diagnosis ; etiology ; Male ; Middle Aged ; Procollagen ; blood

OBJECTIVETo evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms.

METHODSSixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-β1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing.

RESULTSIntraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-β1 and collagen I expressions in the ischemic tissues.

CONCLUSIONSIntraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-β signaling pathway.

Animals ; Cecum ; injuries ; pathology ; Collagen Type I ; metabolism ; Fibrinolysis ; Injections, Intraperitoneal ; Peritoneum ; injuries ; pathology ; Phenanthrenes ; pharmacology ; Plasminogen Activator Inhibitor 1 ; metabolism ; Postoperative Complications ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Tissue Adhesions ; prevention & control ; Tissue Plasminogen Activator ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing