Objective To explore the effects of application of probiotics on intestinal flora and related cytokines in infants with cholestatic liver disease.Methods (1) Eighty-four infants with cholestatic liver disease had been hospitalized from October 2010 to June 2011 in the First Hospital of Guangxi Medical University.The infants with cholestatic liver disease were randomly divided into the probiotic intervention group and the non-probiotic intervention group.Quantification of intestinal bacteria was detected by SYBR Green Ⅰ real-time fluorescent quantitative polymerase chain reaction,then the number of 3 kinds of bacteria before and after the treatment was compared.(2) The indices of liver function,blood ammonia,cholesterol were detected.The levels of serum transforming growth factor-β1,tumor necrosis factor-α(TNF-α)and interleukin-6 (IL-6) were measured by enzyme linked immunosorbent assay.Results (1) The number of bifidobacterium,lactobacillus and bifidobacterium/escherichia coli (B/E) were significantly increased (all P ＜ 0.01),whereas the number of escherichia coli was significantly decreased in the probiotic intervention group(P ＜ 0.05),however,there were no differences in the non-probiotic intervention group(all P ＞ 0.05).(2)The indices of total billirubin,direct billirubin,γ-Glutamyltransferase,total bile acid,alanine aminotransferase,blood ammonia,alkaline phosphatase were significantly improved after therapy in 2 groups (all P ＜ 0.05).The levels of TNF-α and IL-6 were significantly decreased in the probiotic intervention group (t--7.31,P =0.00;t =-2.90,P =0.01),but there were no differences in the non-probiotic intervention group.The level of BA was significantly decreased in the probiotic intervention group than the non probiotic intervention group (t =-8.37,P =0.00).(3) The B/E value were significantly inverse correlated with level of serum IL-6 (r =-0.796,P =0.01).Conclusions It may help to restore the intestinal flora and balance the immune function in infants with cholestatic liver disease after application of probiotics.

ObjectiveTo investigate the safety and efficacy of carbon dioxide ( CO2 ) insufflation during ERCP.MethodsBetween January and August 2011,a total of 102 consecutive patients who underwent ERCP were randomized to accept CO2 insufflation ( n =52 ) of air insufflation ( n =50 ) during the procedure.ERCP was carried out with the same instrument by an expert endoscopist who was blinded to the insufflation gas used and the procedure was controlled at 30 minutes to 1 hour.The heart rate,oxygen saturation of the patient was continuously monitored during the procedure.Before the procedure and 1 hour after the end of operation,abdominal X-ray was taken to evaluate the width of intestine,and the degree of intestinal expansion was defined as normal,mild,moderate and severe according to the width increased.A questionnaire with 100 mm visual analogue scale (VAS) was used to quantify the abdominal pain and distention experienced at 1 hour,2 hours,and 6 hours after the procedure.The patients' vital signs,bowel dilatation,the average operating time,abdominal pain score and distention score on VAS,and complications in 2 groups were analyzed.ResultsThe baseline characteristics of 2 groups were comparable.ERCP was successfully performed in all the patients and no complication was observed.In CO2 group,the average operating time,mean heart rate and oxygen saturation were (45.2 ± 10.6) min,( 102.2 ± 10.3 ) bpm and ( 99.5 ± 0.5)％,which were (48.5 ± 11.2) min,( 100.3 ± 11.4) bpm and (98.9 ±0.6)％,respectively,in air group.There were no significant differences on these items between the 2 groups ( P ＞ 0.05 ).Moderate to severe intestinal expansion 1 hour after ERCP was found in 14 patients (26.9％ ) in CO2 group and in 28 patients (56.0％ ) in air group,and the latter was significantly higher than the former (x2 =11.61,P =0.009).Both of the mean abdominal pain and abdominal distention scores at 1 hour post-ERCP in CO2 group were lower than those in air group,but without significant difference (P ＞0.05).However,the mean abdominal pain scores at 2 hours and 6 hours post-ERCP in CO2 group were significantly lower than those of patients in air group (7.4 ±2.2 vs.18.7 ±4.6 at 2 hours post-ERCP,9.6 ±3.7 vs.20.1 ±4.5 at 6 hours post-ERCP,all P ＜ 0.05 ).Similarly,the mean abdominal distention scores at 2 hours and 6 hours post-ERCP in CO2 group were significant lower than those of patients in air group (7.6 ±3.6 vs.18.3 ±4.1 at 2 hours post-ERCP,8.9 ±3.7 vs.19.4 ±4.2 at 6 hours post-ERCP,all P ＜0.05).ConclusionThe use of CO2 insufflation instead of air during ERCP appears to be safe.Insufflation of CO2 during ERCP palliates the intestinal expansion,post-ERCP abdominal pain and distention comparison to insufflation of air.However,because of the single-center clinical observation with limited number of cases,the safety and efficacy of CO2insufflation during ERCP requires to be further evaluated.

In order to clone and express alcohol dehydrogenase II (ADH2) gene from Saccharomyces cerevisiae in E. coli BL21 (DE3) efficiently, we extracted the total RNA as template and obtained ADH2 gene by RT-PCR and connected ADH2 gene to pTAT plasmids to gain recombinant expression plasmid pTAT-ADH2, then transformed this recombinant expression plasmid pTAT-ADH2 into E. coli BL21 (DE3). The recombinant was induced by IPTG to express ADH2. After purification, ADH2 activity was tested in vitro and toxicologic test was done in mouse. Sequence test showed that the acquired fragments exhibited 90% homology to ADH2 gene sequence from GenBank report. The target gene expressed efficiently and took up to approximant 50% of total protein by SDS-PAGE and band scanning analysis. The purified protein exhibited the identified activity through biochemical test and mouse toxicological test. As a result, the acquired ADH2 gene was highly homology to the published sequence and expressed at a high level in E. coli BL21 (DE3), more importantly, ADH2 proved to have ethanol dehydrogenase activity.
Alcohol Dehydrogenase ; genetics ; isolation & purification ; Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Mice ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Saccharomyces cerevisiae ; enzymology ; genetics ; Saccharomyces cerevisiae Proteins ; genetics ; isolation & purification