Objective To analyze the genetic characterization(evolution, antigenicity, enzyme activity sites and glycosylation sites)of the neuraminidase(NA)gene of the novel influenza virus A/H1N1 in 2009 pandemic in Guangdong Province. Methods The viral RNA was extracted from 69 isolates of influenza virus A/H1N1 from patients in 2009 pandemic in Guangdong Province. NA gene fragments were amplified by reverse transcription polymerase chain reaction (RT-PCR) and sequenced. The other 52 NA gene sequences of influenza virus A in different years and different regions were retrieved from GenBank. The analysis of evolution and amino acid sequences were analyzed by MEGA 4.0 software. Results The homology of 2009 novel H1N1 influenza viruses in Guangdong and avian H5N1 influenza virus strains was high(＞85 % ). The amino acid distributions of potential antigenic sites were identical. The enzyme activity sites of NA genes of all virus strains were strictly conserved, which had eight glycosylation sites. But there were amino acid substitutions in 5 glycosylation sites, while it was identical with the 2001 avian H5N1 influenza virus. Conclusion The NA genes of 2009 novel H1N1 influenza viruses in Guangdong are high homologous with avian H5N1 influenza virus and the viral specific binding sites of neuraminidase inhibitor are not changed.

Objective To explore the clinical efficacy of laparoscopic left hemihepatectomy for the treatment of intrahepatic bile duct stones.Methods The clinical data of 30 patients with left intrahepatic bile duct stones who were admitted to the Second Affiliated Hospital of Nancbang University from June 2013 to June 2014 were retrospectively analyzed.All the patients underwent laparoscopic left hemihepatectomy by the Glisson intra-and extra-pedicles vascular inflow occlusion techniques together with the removal of choledocholithiasis and right bile duct stones,and T tube placement or laparoscopic primary suture of common bile duct were selected according to the condition of bile duct.All the 30 patients were readmitted to hospital and detected by color Doppler ultrasound (CDUS),computed tomography (CT) and T tube cholangiography at postoperative month 1,and then received CDUS reexamination every 3 months.CT and MRI reexaminations were applied to patients with complication of residual stones if necessary.All the patients were followed up till July 2014.Results All the 30 patients were treated by laparoscopic hepatectomy with left hemihepatic vascular inflow occlusion,including 5 with conversion to open surgery and 25 with successful operation.The Glisson extra-and intra-pedicel vascular inflow occlusion techniques were used in 11 and 14 patients,respectively.The operation time and volume of blood loss were (158 ± 85) minutes and (405 ± 215) mL.Two patients received intraoperative blood transfusion.There were no residual stones in the 8 patients with choledocholithiasis by intraoperative choledochoscope,and primary suture of bile duct and T tube placement were done in 5 and 3 patients,respectively.No patients died.After operation,there were 2 patients with bile leakage and 1 with pleural effusion,and they were cured though drainage.One patient with subphrenic effusion was cured by B ultrasound-guided puncture and drainage.One patient had bleeding with the volume of blood loss of 500 mL,and was cured by conservative treatment.The duration of hospital stay in all the patients was (8.5 ± 2.3)days.No bile leakage and abdomen infection were detected by outpatient examination.The time of followup was 1-12 months,without recurrence of stones.Conclusion Laparoscopic left hemihepatectomy for the treatment of left intrabepatic bile duct stones is safe and feasible with satisfactory outcome.

. Conclusions Two multiplex RT-PCR assays show high consistency with common RT-PCR. The multiplex RT-PCR assays were initially established.

Objective To investigate the expression and clinical significance of LINC00638 in colon cancer,and further to explore the impact of LINC00638 on the biological function of colon cancer cells.Methods The cancer genome atlas(TCGA)database was used to preliminarily compare the expression differences of LINC00638 in normal colon tissue and colon cancer tissue,and to explore the rela-tionship between LINC00638 and clinical pathological characteristics of patients.Kaplan-Meier survival analysis was used to further in-vestigate the effect of LINC00638 expression on the prognosis of colon cancer patients.The GEPIA database and the Gene Expression Om-nibus(GEO)database were used to further validate the effect of LINC00638 expression on the prognosis of colon cancer patients.Univari-ate and multivariate COX regression analysis were used to explore the effect of LINC00638,clinical and pathological related factors on the prognosis of colon cancer patients.Small interfering RNA(siRNA)was used to silence the expression of LINC00638 in HCT116 cells and DLD1 cells,and the effect of LINC00638 on the proliferation,migration and invasion of colon cancer cell was explored through CCK-8 experiment,colony formation experiment,scratch experiment and Transwell experiment.Results The expression level of LINC00638 was significantly increased in colon cancer tissue,and its expression level was closely related to lymph node metastasis and TNM staging in co-lon cancer patients.The results of Kaplan-Meier survival curve analysis showed that the higher the expression level of LINC00638,the worse the prognosis of colon cancer patients(P＜0.05).The results of COX regression analysis showed that LINC00638,age,and TNM staging could be independent prognostic factors for colon cancer patients.The results of cell function experiments showed that compared with the control group,the proliferation,migration,and invasion ability of cells in knockdown group were significantly decreased(P＜0.05).Conclusion LINC00638 is highly expressed in colon cancer,and the higher its expression level,the worse the prognosis of colon cancer patients.After knocking down LINC00638,it can inhibit the proliferation,migration and invasion ability of colon cancer cells.

Objective To understand the evolutionary characterization of hemagglutinin (HA)gene of pandemic H1N1 influenza virus in Guangdong during 2009-2011. MethodsWe selected 83 pandemic H1N1 strains isolated in Guangdong during 2009-2011. The HA1 genes were sequenced and analyzed comparatively by Bioedit 7.0 and MEGA 4.0. ResultsThe evolutionary rate of Hal gene of pandemic H1N1 and seasonal H1N1 viruses was 5.2×10-3 substitutions/site/year, higher than that of seasonal H1N1 viruses. Most amino acid changes in HA1 molecules accumulated on the surface of the molecule and were partly located in antigenic sites. Two fatal infections were detected with a mutation at HA residue 222, in one virus with a change D222G, and in one virus D222N. ConclusionThe phylogenetic analysis demonstrates that the influenza epidemic in Guangdong at the beginning of 2011 are due to occurrence of genetic changes of pandemic H1N1 virus. The amino acid change at residue 222 of the HA1 are likely to be associated with severe or even fatal illness.

Objective To investigate the susceptibility to oseltamivir of influenza pandemic A (H1N1) 2009 viruses in Guangdong province during 2009,provide valuable information of prescription for clinics,and elucidate the variant trend of the epidemic strain based on phylogenetic analysis.Methods During April to December 2009,clinical specimens were collected from sentinel hospitals covering the whole Guangdong province.Virus isolation was performed by in MDCK cells or embryonated chicken eggs.A fluorescence-based neuraminidase (NA) enzyme inhibition assay was conducted to measure influenza susceptibility.The NAI susceptibility of influenza virus was expressed as the concentration of NAI needed to inhibit the NA enzyme activity by 50％.A subset of 68 viruses were performed NA sequencing for detecting resistant mutations and studying variant trends.Results During surveillance,221 influenza pandemic A ( H1N1 ) viruses were isolated.All strains were sensitive to oseltamivir inhibition assay,with a median IC50 of 0.24 nmol/L (range 0.02 -1.66 nmol/L).No mutation related to resistance was found.Phylogenic analysis illustrated that these NA genes were homology high to 99.5％ - 100.0％ with those from other countries.Conclusions influenza pandemic A (H1N1) 2009 viruses were sensitive to oseltamivir in Guangdong,and useful for prophylaxis and treatment of influenza infection.Little selective pressure was found by phylogenic analysis.Our laboratory will continue to observe antiviral-resistance among circulating influenza viruses.

Objective To investigate the efficacy of amoxicillin,clavulanate potassium and cefotaxime in the treatment of neonatal pneumonia and its influence on intestinal microflora.Methods 310 cases of neonatal pneumonia diagnosed in the hospital from June 2014 to June 2016 were selected as the subjects.They were divided into three groups according to random number table.The group A (100 cases) was treated with amoxicillin and clavulanate potassium,group B(105 cases) was treated with cefotaxime,group C(105 cases) was treated with amoxicillin,clavulanate potassium and cefotaxime.The effect and the difference of intestinal microecology were compared among the three groups.Results There were no statistically significant differences among the three groups in effect and the cure time [(5.2 ± 1.3) d vs.(5.4 ± 1.5) d vs.(5.3 ± 1.4) d] (P ＞ 0.05).There were statistically significant differences in intestinal microecology among the group A,group B,group C and control group (x2 =11.741,7.066,6.163,8.752,22.851,all P ＜ 0.05).The lactobacillus,Bifidobacterium,Enterococcus and true Bacillus of group A,group B,group C were less than those of the control group,which in group C were significantly less than group A and group B,there were statistically significant differences (t =10.453,6.843,5.974,17.654,all P ＜ 0.05).The Enterobacteriaceae of group A,group B and group C was significantly more than that of the control group,and the Enterobacteriaceae of group C was significantly more than that in group A and group B,there were statistically significant differences (t =6.574,4.634,3.968,9.893,all P ＜ 0.05).Conclusion Amoxicillin,clavulanate potassium and cefotaxime are effective in the treatment of neonatal pneumonia,the combined use of drugs has great influence on intestinal microflora.

To investigate the genetic character and origin of the first imported infection case of middle East respiratory syndrome coronavirus (named as MERS-CoV_China GD01), RNA was extracted from swabs of this patient followed by RT-PCR amplification. All coding gene of structural (S, E, M, E) and accessory (ORF3, ORF4a, ORF4b, ORF5, ORF8b) proteins were sequenced and analyzed. Phylogenetic analyses of structural protein coding genes of MERS-CoV_ China GD01 indicates that several substitutes exists in S coding gene and its origin belong group 5 of MERS-CoV, which were recent circulated in Saudi Arabia area, while other three structural genes (N, E, M) were very conserved. Phylogenetic analyses of accessory protein coding genes of MERS-CoV China GD01 indicates that several substitutes exists among ORF3, ORF4a, ORF4b and ORF5, while ORF8b was conserved. In conclusion, genome of MERS-CoV_ China GD01 was general conserved although several genetic variations were found among structural and accessory protein coding genes. This is the first report on sequencing and phylogenetic analyses of the first imported MERS case in China, which may pay the way for prevention and control of imported MERS-CoV infection.
China ; Conserved Sequence ; Coronavirus Infections ; transmission ; virology ; Evolution, Molecular ; Genomics ; Humans ; Middle East Respiratory Syndrome Coronavirus ; genetics ; physiology ; Phylogeny ; Sequence Analysis ; Viral Proteins ; genetics

OBJECTIVETo assess the effect of cholesterol in bile on cholecystokinin receptor (CCK-R) in the gallbladder.

METHODSOne hundred Guinea pigs were randomly divided into four groups, 25 animals for each. The control group was fed a standard diet, and the cholesterol group fed a diet containing 2% cholesterol. After taking the 2% cholesterol diet for two weeks, the natural group persisted on the standard diet, and the treated group was perfused by traditional Chinese medicine. Serum cholecystokinin (CCK) level in the portal vein and maximal binding capacity (B(max)) and Kd of CCK-R in the gallbladder were measured in the four groups by RIA and RBA, and the concentrations of cholesterol in bile were also observed.

RESULTSCompared with the control group, after high-cholesterol feeding for two weeks, the gallbladder emptying rate [(65.83 +/- 7.32)% approximately (47.22 +/- 5.24)%] and B(max) of CCK-R [(60 +/- 27) approximately (32 +/- 13) fmol/mg protein] and in decreased fasting gallbladder volume (FV) [(0.89 +/- 0.26) approximately (1.34 +/- 0.61) cm(3)] and concentration of cholesterol [(0.44 +/- 0.11) approximately (0.60 +/- 0.13) mmol/L] in bile increased, but no change was in the serum CCK level and Kd of CCK-R in the cholesterol group. Compared with the natural group, after two-week in take of herb decoction of qingre lidan and liqi huoxue, FV [(1.27 +/- 0.60) approximately (0.90 +/- 0.27) cm(3)], RV [(0.85 +/- 0.45) approximately (0.32 +/- 0.12) cm(3)], FB [(0.92 +/- 0.35) approximately (0.73 +/- 0.21) cm(3)], RB [(0.76 +/- 0.34) approximately (0.29 +/- 0.08) cm(3)] in the treated group decreased significantly; but gallbladder emptying rate [(43.06 +/- 4.27)% approximately (67.01 +/- 6.82)%] increased significantly. The concentration of cholesterol in bile was lower in the treated group than in the natural group [(0.59 +/- 0.14) approximately (0.43 +/- 0.10) mmol/L], but no change was found in the serum CCK level. Bmax of CCK-R in the treated group increased significantly [(39 +/- 19) approximately (59 +/- 11) fmol/mg protein], Kd of CCK-R showed no significant changes between the treated group and natural group.

CONCLUSIONHigh cholesterol in gallbladder bile causes defective muscle contraction by down-regulating CCK-R in the gallbladder, so the reduction of cholesterol concentration of bile may contribute to gallbladder contraction.

Animals ; Bile ; chemistry ; Cholecystokinin ; blood ; Cholesterol ; analysis ; physiology ; Gallbladder ; chemistry ; physiology ; Guinea Pigs ; In Vitro Techniques ; Male ; Muscle Contraction ; Receptors, Cholecystokinin ; analysis

Objective To detect the positive rates of antibodies against avian influenza virus (AIV) subtypes H5, H6, H7 and H9 among people in poultry occupations in Guangdong province and to analyze the transmission of various subtypes of AIV from poultry to human contacts for the prevention and control of novel AIV infection in human beings.Methods Serum specimens were collected from 1066 peo-ple in poultry occupations ( occupational group) and 205 people not in poultry occupations ( non-occupational group) in 10 cities of Guangdong province.The inactivated AIV strains, isolated from poultry or environment of Guangdong province, were used as antigens to detect antibodies against AIV subtypes H5, H6, H7 and H9 by using the hemagglutination inhibition ( HI) assay.Results The positive rates of antibodies against AIV subtypes H5, H6, H7 and H9 carried by people from the occupational group were respectively 0.44%, 0%, 0.30%and 0.30%in 2013 and 1.08%, 0.0%, 0.0%and 0.27%in 2014.Only the anti-H9 anti-bodies were detected in serum samples collected form people in the non-occupational group in 2013 with a positive rate of 0.95%.No significant differences with the positive rates of anti-AIV antibodies were found between the occupational group and the non-occupational group.However, the geometric mean titer ( GMT) of anti-AVI antibodies in people from the occupational group was higher than that of the non-occupational group.Conclusion Although a grand spread of AIV from avian to human is not likely to happen yet, con-tacting with poultry is the risk factor for AIV infection in Guangdong population.A long-term surveillance of anti-AIV antibodies in serum should be strengthened among people in poultry occupations for the timely pre-vention and control of novel AIV outbreak.