Objective To identify the pathogen of an influenza epidemic situation and analyze the genetic characteristic of hemagglutinin( HA ) gene and neuraminidase(NA) gene of this pathogen. Methods Real-time RT-PCR was used to dectect nucleic acid of the pandemic( H1N1 ) 2009 virus from oropharyngeal swabs of initial influenza-like illness in epidemic. The viruses were was inoculated and isolated with embryonated eggs. And the HA gene and NA gene were sequenced to analyze their characteristic. Results The influenza epidemic situation was caused by the pandemic( H1N1 ) 2009 virus. The HA and NA sequences data showed that the virus had the high homology with reference virus, and the NA sequences had not the H274Y mutation. Conclusion In this study, the pandemic( H1N1 ) 2009 virus were similar with the vaccine-like virus and the isolated virus of China, and sensitive to oseltamivir.

AIM:To study the effect of proprotein convertases (PCs) on the transforming growth factor (TGF) ?1-induced inhibition of HBV replication.METHODS:HepG2.2.15 cells cultured regularly were exposed to recombinant TGF?1 at concentration of 2 ?g/L or 5 ?g/L and/or PC inhibitor at concentration of 20 ?mol/L for 18 h.The total RNA and HBV core particle DNA were extracted from these cells,and PC mRNA and core-associated HBV DNA were detected by real-time PCR technique.RESULTS:The mRNA expression levels of 7 PCs in HepG2.2.15 cells were observed with various degrees.Recombinant TGF?1 significantly up-regulated the mRNA expression of all PCs except for the down-regulation of PC5 /6,though PC1 /3 and PC2 were up-regulated most obviously.Furin and PACE4 were the predominant PCs before and after TGF?1 exposure when the basic mRNA expression was taken into account.Further study showed that TGF?1-induced the inhibition of HBV replication was abrogated by PC inhibitor in HepG2.2.15 cells.CONCLUSION:TGF?1-induced the inhibition of HBV replication is mediated by the up-regulation of PCs,which might be of many implications in efficient interferences of TGF?1 on HBV replication.

Objective To explore the influence of intestinal trefoil factor (ITF) on interleukin-6(IL-6) in neonatal rat with necrotizing enterocolitis(NEC) ,and to discuss the protective machanism of ITF on NEC.Methods Thirty-two neonatal rats were divided randomly into four groups,group A as control group,group B as NEC group,group C as NEC+NS 0.2 mL group,group D as NEC+ITF 0.2 mg group.NEC model of neonatal rats were established.On the 4th day,all the subjects were put to death.Intestinal tissue within the boundary of ileum and cecum was obtained to observe histological changes.Other intestinal tissue was treated into homogenate.After the homogenate was centrifuged,supernates were used to test the density of IL-6.Results The density of IL-6 significantly decreased in group A,D than those in group B and C (Pa0.05).The pathological lesions indicated that intestinal tissue necrosis was severe in group B and C,which was graded as 3 points,but obviously lessen in group D,which was graded as 1 point,with ITF interfering.Conclusions Intestinal inflammation is ameliorated after ITF are injected hypodermically or intraperitoneally.ITF may provide a brand-new way for the therapy of NEC in neonatal rats.

Objective:To search, evaluate and integrate the best evidence for lactation maintenance of very low birth weight infant(VLBWI) mothers, and to provide evidence for lactation maintenance.Methods:Up to Date, BMJ best practice, Newborn Services Clinical Guidelines, ABM, NICE, OVID evidence-based database, NGC, Joanna Briggs Institute Library, Cochrane Library, Web of Science, PubMed, CNKI, CBM and Wanfang Data were retrieved by computer on the Maintenance of Mother′s Lactation in VLBWI: Guidelines, Systematic Evaluation, Expert Consensus/Opinions, Evidence Summary Literature screening, quality evaluation, evidence extraction and integration of documents which met the requirements.Results:A total of 10 articles were included, including 3 guidelines, 4 systematic reviews, 1 expert consensus and 2 expert opinions. A total of 41 evidences were summarized from 9 aspects, including lactation support, lactation initiation, lactation monitoring and the application of lactating agents.Conclusion:The evidence of lactation maintenance of VLBWI mothers is sufficient, which can provide a basis for medical staff to make clinical decisions.

Objective To investigate the number and activities changes of peripheral blood endothelial progenitor cells (EPCs) in continuous ambulatory peritoneal dialysis (CAPD) patients,and explore the connection between EPCs' number and the levels of advanced glycosylation end products (AGEs),homocysteine (Hcy) and C-reactive protein (CRP).Methods Twenty-five CAPD patients and thirty healthy volunteers were involved.Total mononuclear cells (MNCs) were isolated from peripheral blood of patients.EPCs were characterized as adherent cells by double staining of FITCUEA-1 and DiL-AcLDL binding,and were further demonstrated by positive cells of CD34,CD133 and KDR using flow cytometry.The abilities of cell proliferation,adhesion and migration were further observed by fluorescent microscope.The correlations between the CEPCs' number and the levels of AGEs,Hcy and CRP were analyzed.Results The number and activities including migration and adhesion of EPCs in CAPD group were significantly lower than control group (P ＜ 0.05).The levels of serum AGEs,Hey and CRP in CAPD patients were increased (all P＜0.05) and had negative correlation with EPCs' number.Conclusions The number and activities of EPCs decrease in patients with CAPD,and EPCs' number is negatively correlated to the levels of AGEs,Hcy and CRP.

Background Our previous study demonstrated that the aptamer S58 specifically targeted transforming growth factor-β receptor Ⅱ (TβRⅡ) and inhibited the transdifferentiation of human Tenon capsule fibroblasts (HTFs) mediated by transforming growth factor-β (TGF-β).Chitosan-nanoparticles (CS-NP) are good drug carriers,but the efficacy and safety of CS-NP/aptamer complexes deserve attention.Objective The aim of this study was to synthesize a novel CS-NP/aptamer complex called CS (S58)-NP and investigate its properties and applicability.Methods Human Tenon capsule tissue was obtained from patients during strabismus surgery,and HTFs were cultured and passaged using the explant culture method.The fourth to tenth generations of cells were used in the experiment.Different concentrations of CS-NP were used to prepare the CS(S58)-NP by the ionic cross-linking method with a surface charge rate (N/P) for S58 of 10,20,30 or 40.The particle size and Zeta potential were measured by the Zeta analyzer.The shape and distribution of CS (S58)-NP particles were examined under the scanning electron microscope.The binding of CS-NP with S58 and resistance of CS (S58)-NP to DNase Ⅰ were examined by agarose gel eletrophoresis.The release rate of S58 from CS (S58)-NP in PBS was quantitatively analyzed by a ultraviolet spectrophotometer.The cytotoxicity of CS(S58)-NP to HTFs was evaluated by detecting the production of lactate dehydrogenase (LDH).Results The Zeta analyzer showed that the particle size of CS (S58)-NP was 130-270 nm and its electric potential ranged from + 16 to +28 mV.The CS (S58)-NP particles appeared spherical with an even distribution under the scanning electron microscope.The mean encapsulation efficiency of CS(S58)-NP was 88.9％,89.3％,91.7％ or 90.5％,respectively,when the N/P was 10,20,30 or 40.After being encapsuled by CS-NP,S58 could resist the degradation from DNase I.Its total releasing level in PBS increased with the lapse of time,with a maximum releasing speed at 24 to 36 hours.The total releasing level reached 100％ at 96 hours.With increaseing concentrations of CS(S58)-NP,the relative releasing level of LDH in HTFs suspension gradually elevated with a significant difference among the groups (F =588.018,P =0.000),with the highest released LDH level at 50 nmol/L of CS(S58)-NP (12.853％ ±0.375％).Conclusions CS-NP provides a protective and slow-releasing effect on the S58 aptamer.CS (S58)-NP shows a good biocompatibility with HTFs with a low cytotoxicity at a concentration of ＜50 nmol/L.CS(S58)-NP could be used to inhibit TGF-β induced transdifferentiation of HTFs in the future.

OBJECTIVETo search for an effective method for treatment of obstinate hiccup.

METHODSNinety cases of obstinate hiccup secondary to cerebral diseases were randomly divided into 3 groups, 30 cases in each group. Group A were treated by injection of compound chlorpromazine into Yintang (Ex-HN 3), group B by intramuscular injection of compound chlorpromazine and group C by acupuncture at Yintang (Ex-HN 3). Their therapeutic effects were compared.

RESULTSThe effective rate was 93.3% in the group A, 10.0% in the group B and 30.0% in the group C, the group A being better than the group B and the group C (P < 0.01).

CONCLUSIONInjection of compound chlorpromazine into Yintang (Ex-HN 3) has a significant therapeutic effect on obstinate hiccup.

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.
Cluster Analysis ; DNA, Fungal ; genetics ; Endophytes ; classification ; genetics ; physiology ; Fusarium ; classification ; genetics ; physiology ; Phylogeny ; Plant Leaves ; metabolism ; Thymelaeaceae ; microbiology ; Wood ; metabolism ; microbiology

Objective To investigate the overall survival prediction of the Bolondi substaging model for patients in intermediate-stage of Barcelona clinic liver cancer (BCLC) after hepatectomy.Methods The retrospective cohort study was adopted.The clinical data of 343 patients with intermediate-stage hepatocellular carcinoma (HCC) who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University between February 2008 and January 2010 were collected.All the patients received the detailed medical history collection,physical examination,laboratory and imaging examinations after admission,and then hepatectomy was performed according to the results of above examinations.Research methods:(1) patients were allocated into the B1,B2 and B3/4 groups based on the Bolondi's substaging model,and the prognostic analyses among groups were conducted.(2) The related factors affecting the prognosis of patients in the B1 and B2 groups were analyzed.(3) The patients in the B1 and B2 groups were allocated into the 4 groups [patients of B1 group with negative microvascular invasion (MVI) were divided in the M1 group,patients of B1 group with positive MVI in the M2 group,patients of B2 group with negative MVI in the M3 group and patients of B2 group with positive MVI in the M4 group] according to the situations of MVI,and stratified analysis was conducted.Observation indicators:basic clinical and pathological features and survival of patients in the B1,B2 and B3/4 groups were observed.Risk factors analysis affecting the prognosis of patients and stratified analysis of MVI in the B1 and B2 groups were conducted.All the patients were followed up by outpatient examination and telephone interview up to February 2014,and the abdominal ultrasound,liver function and serum alpha-fetoprotein (AFP) tests was performed once every 3 months within 2 years postoperatively and once every 6 months after 2 years postoperatively.The continuous variables and categorical variables were respectively represented as M(Qn) and percentage.The comparisons of continuous variables and categorical variables among groups were analyzed by ANOVA or Kruskal-Wallis test and chi-square test or Fisher exact probability,respectively,and one-way ordinal categorical variables were analyzed by the Kruskal-Wallis test.The survival curve was drawn using the KaplanMeier method.The univariate analysis and multivariate analysis were done using the Log-rank test and COX regression model.Results (1) The basic clinical pathological features:of 343 patients with HCC,143,183 and 17 patients (12 in the B3 substaging and 5 in the B4 substaging) were respectively allocated into the B1,B2 and B3/4 groups.There were statistically significant differences in the age,peritoneal effusion,total bilirubin (TBil),albumin (Alb),alanine transaminase (ALT),prothrombin time (PT),platelet (PLT),alpha-fetoprotein (AFP),extent of liver resection,surgical margin ivasion,tumor diameter,number of tumor,Edmondson-Steiner grade,Up-to-7 score,Up-to-7 standard and Child-pugh score among the 3 groups (F =3.377,NA,11.245,32.616,6.884,11.564,33.100,12.902,NA,NA,239.089,10.357,x2=8.906,F =251.508,x2 =343.000,106.790,P ＜ 0.05).(2) Survival of patients:all the patients were followed up for 2.8-70.8 months with a median time of 38.7 months.The 1-,3-,5-year survival rates and median survival time in the B1,B2 and B3/4 groups were 85.8％,72.8％,52.9％ and 63.2％,47.5％,16.8％ and 45.5％,30.4％,8.4％ and 55.1 months,35.1 months,12.2 months,respectively,showing a statistically significant difference (x2 =22.800,P ＜ 0.05).(3) Risk factors analysis:the results of univariate analysis showed that the peritoneal effusion,Alb,Hb,AFP,esophagogastric varices,surgical margin invasion,tumor diameter,MVI and Edmondson-Steiner grade were related risk factors affecting the prognosis of patients with HCC after hepatectomy [HR =2.04,2.46,2.50,1.78,1.55,3.54,1.71,1.76,1.69,95％ confidence interval (CI):1.13-3.69,1.20-5.02,1.51-4.15,1.29-2.45,1.06-2.25,1.65-7.61,1.23-2.38,1.23-2.51,1.08-2.64,P＜0.05].The results of multivariate analysis showed that the Alb ＜ 35 g/L,Alb ＜ low limit of normal,tumor invading to surgical margin,tumor diameter ＞ 5 cm and positive MVI were independent risk factors affecting the overall survival of patients with HCC after hepatectomy (HR =2.82,2.16,2.93,1.48,1.53,95％ CI:1.37-5.80,1.27-3.69,1.33-6.44,1.05-2.09,1.06-2.22,P＜0.05).(4) There were 61,82,57 and 126 patients in the M1,M2,M3 and M4 groups,and M2 and M3 groups were merged into the M2/3 group because of being similar survival situations of patients.The 1-,3-,5-year survival rates and median survival time in the M1,M2/3,and M4 groups were 90.0％,83.2％,67.7％ and 68.8％,59.9％,41.6％ and 52.7％,42.1％,23.6％ and 69.0 months,49.2 months,24.9 months,respectively,with a statistically significant difference among the 3 groups(x2=20.200,P ＜ 0.05).Conclusions The Bolondi substaging model produces an optimal survival prediction for patients in intermediate stage of BCLC after hepatectomy.The patients in the B1 and B2 substaging have better long-term survival outcomes after hepatectomy.

Objective To investigate the changes of the N-acetylaspartate(NAA) concentrations in different brain regions and executive function skills in alcohol dependence,and to study the relationship between NAA levels and cognitive functions in subjects.Methods 49 male,non-smoking,alcohol-dependent patients and 45 healthy control subjects were measured with Proton 1H Magnetic resonance spectroscopy (MRS) and Wisconsin Card Sorting Test (WCST).Results Alcoholics had lower NAA/Cr ratios in prefrontal grey matter(GM) (1.59± 0.13) and white matter(WM) (1.58±0.12) regions and performed poorly on executive function tests compared to controls (P＜0.001).NAA/Cr in left prefrontal regions positively correlated with certain parameters of EF testing (number of correct responses 30.37± 3.73,perseverative errors 11.49± 3.39,random errors 6.18± 2.64,categories completed 2.08± 1.59)in alcoholic group (P＜0.01).NAA/Cr in prefrontal WM regions correlated with certain parameters of EF testing in alcoholic group (number of correct responses r=0.379,categories completed r=0.433,P＜ 0.05).Conclusion Long-term,chronic alcoholism will damage neuronal viability and cognitive functions,which suggests that NAA concentrations can reflect the extent of damage of cognitive functions with decreased levels reflecting neuronal loss.