Objective To construct a ciaR gene-knockout (ΔciaR) mutant of Streptococcus pneu-moniae ( S. pneumoniae) and to investigate the effects of CiaR in CiaH/CiaR, a streptococcal two-component signal-transducing system, on the expression of genes encoding penicillin-binding proteins ( pbps genes) and cia-dependent small RNAs (csRNAs). Methods Electrophoretic mobility shift assay (ESMA) was per-formed to detect the recombinant CiaR (rCiaR)-binding pbps genes. A suicide plasmid pEVP3ciaR for ciaR gene knockout was constructed and then aΔciaR mutant was obtained through homologous recombination and insertion inactivation of the suicide plasmid, and screening with chloromycin. The mutant was identified using PCR and sequencing analysis. E-test was used to detect the minimal inhibitory concentrations ( MIC) of penicillin ( PCN) and cefotaxime ( CTX) against S. pneumoniae strains. Changes and differences in the expression of pbps genes and csRNAs in theΔciaR mutant and its wild-type strain before and after treatment with 1/4 MIC of PCN or CTX were detected using real-time quantitative RT-PCR. Results The rCiaR could bind to the promoter regions in pbp1a, pbp1b and pbp2b genes of S. pneumoniae. The ciaR gene in ΔciaR mutant was inactivated by insertion according to the results of PCR and sequencing analysis. After treatment with 1/4 MIC of PCN or CTX, the expression of pbps genes at mRNA level ( pbps-mRNAs) in theΔciaR mu-tant was significantly increased (P<0. 05), but the levels of csRNAs were significantly decreased (P<0. 05);whereas a significantly decreased pbps-mRNAs (P<0. 05) and increased csRNAs (P<0. 05) were observed in its wild-type strain. The result of E-test showed that the MICs of PCN and CTX against ΔciaR mutant were increased by 250-fold as compared with those against its wild-type strain. Conclusion The CiaR can enhance the drug resistance of S. pneumoniae to PCN and CTX through down-regulating the expres-sion of PBP1a, PBP1b and PBP2b and up-regulating the expression of csRNAs to inhibit the expression of PBPs.

Objective:To study the Fanggan Decoction,s death prevention on mouse and inhibition effects on Influenza A virus in vivo.Methods:After setting up the model of mouse infected with Influenza A virus(H1N1),we observed the death prevention with Fanggan Decoction,done hemagglutination test and detected the dynamic contents of virus with Real-Time Fluorescence Quantitative RT-PCR.Results:Fanggan Decoction can prevent the death of infected mouse and delay the survival time.The death rate was 66.67%,33.33% and 25% respectively in low,middle and high dose of Fanggan Decoction groups and the average survival time was respectively 8.75 days,11.41 days and 12.33 days.Virus contents reached peak on the 5th day,while compared with the model group,virus contents were lower in each Fanggan Decoction groups,especial in the middle and high dose groups.Conclusion:Fanggan Decoction had good effect in inhibiting Influenza A virus,and can prevent the death of infected mouse,delay the survival time,while get better antivirus dose-effect relationship at double dose.

Objective To evaluate the value of serum procalcitonin (PCT ) in the diagnosis of liver cirrhosis complicating sponta‐neous bacterial peritonitis (SBP) .Methods The patients with cirrhosis were divided into non‐SBP group and complicating SBP group according to whether complicating SBP ,and the patients with common hepatitis served as the control group .Serum expres‐sion levels of PCT ,CRP and IL‐6 were detected by electrocheniluminescence and Immunoturbidimetry methods .The receiver operat‐ing characteristic curve (ROC) curve was drawn for evaluating the diagnostic efficiency of each indicator .Results The levels of ser‐um PCT ,CRP and IL‐6 in the complicating SBP group were significantly higher than those in the non‐SBP group and control group , the differences were statistically significant (P<0 .01);The ROC curve analysis showed that the diagnostic value of PCT at the op‐timum threshold value of 0 .51 ng/mL for diagnosing cirrhosis complicating SBP was superior to CRP and IL‐6 ,the sensitivity of se‐rum PCT for diagnosing SBP in 3 groups was 62 .68% ,the specificity was 76 .59% and the accuracy was 80 .01% .Conclusion The serum PCT level has an important value for the early diagnosis of liver cirrhosis complicating SBP .

ObjectiveTo evaluate the feasibility, safety and the long-termoutcomes of laparoscopy-assisted distal gastrectomy (LADG) for advanced gastric cancer (AGC).MethodsWe retrospectively analyzed the clinical and follow-up data of 346 cases after LADG from January 2004 to June 2009, compared with 313 cases after conventional open distal gastrectomy (ODG) for advanced gastric cancer at the same period at our hospital. The surgical safety, postoperative complications, survival rate, and the recurrence and metastasis of cancer were compared.ResultsThere was no significant difference at the average time of LADG and ODG procedures (211 ± 56) min vs.(204 ±41 ) min, but blood loss during operation and length of incision in LADG group were significantly less than in the ODG group. The proximal and distal length were, respectively, (6. 3 ± 2. 0) cm and (5. 7 ± 1.7 ) cm in LADG group and (6. 3 ±2. 1 ) cm and (5.6 ± 1.6) cm in ODG group, the difference was not significant. The number of lymph node dissections was also similar: (33 ± 13) in LADG group and (33 ± 16) in ODG group. The incidence of postoperative complications in LADG group was significantly lower than that in ODG group ( 6. 7％ vs.13. 1％, P ＜ 0. 05). During the follow-up period of 6-72 months (average 37 months), the 1-, 3-and 5-year survival rates were, respectively, 87. 2％, 57. 2％ and 50. 3％ in LADG group and 87. 1％, 54. 1％and 49. 2％ in ODG group, the difference was not significant. The differences in recurrence and metastasis between the two groups were not statistically significant.ConclsionLaparoscopy-assisted gastrectomy for advanced gastric cancer is not significantly different with open surgery in postoperative survival rate or recurrence. It is less traumatic and of fewer complications.

ObjectiveTo evaluate the diagnostic value of CT in adults with midgut malrotation.MethodsThe CT and clinical findings of 13 cases confirmed by surgery were analyzed retrospectively.ResultsCT found “whirl sign” in 9 cases,“transposition sign” in 5 cases,and the superior mesenteric vein located in right lower abdomen in 4 cases.At the same time,duodenum and upper jejunum located in right upper abdomen.Conclusion“Whirl sign” and “transposition sign” were the typical CT findings of midgut malrotation in adults.Ectopic superior mesenteric vein highly indicated slight midgut malrotation in adults.

Objective To investigate the value of remnant stomach-jejunal dual pathways reconstruction after laparoscope-assisted radical proximal gastrectomy in the treatment of upper gastric cancer. Methods Twenty-five patients with upper gastric cancer underwent laparoscope-assisted radical proximal gastrectomy and the remnant distal stomach was preserved for side-to-side remnant stomach-jejunal anastomosis and end-to-side jejuno-jejunal anastomosis to reconstruct dual pathways. Results The mean operation time was (240±35) minutes, the mean number of lymph nodes dissected were 22±5, and all the incised margins were negative. No anastomotic leakage, obstruction or stenosis occurred. All patients received postoperative barium meal examination. A large amount of barium directly entered the jejunum, leaving a small amount of barium entered the jejunnum via the route of remnant stomach-duodenum, and was detained in the remnant stomach for 30-60 minutes. No esophageal reflux of barium was observed. All the patients were followed up for 4-18 months, no reflux esophagitis was detected and the short-term life quality was satisfactory. Conclusions Remnant stomach-jejunal dual pathways reconstruction prevents the reflux esophagitis and dumping syndrome, preserves the pathway of duodenum and promotes the life quality of patients.

Objective To investigate the feasibility,method and result of laparoscopic total gastrectomy for gastric cancer. Methods Clinical data of 63 cases of gastric cancer treated with laparoseopic total gastrectomy were analyzed retrospectively. Results In this study,52 cases underwent laparoseopic radical total gastrectomy and 5 cases did laparoseopic palliative total gastrectomy.The procedure Was hand assisted in tlle first 45 cases for fashoning esophagojejunostomy through a small incision.In six cases the procedure was converted to open surgery.The operative time was(312±35)min,the blood loss was(190±50)ml,the number of lymph nodes dissected Was(32±7).It began to pass flatus(4.0±1.2)days postoperatively.It was(4.5±1.5)days to start oral liquids.Patients were up and about on(4.0±1.5)days postoperatively. Minor postoperative complications occurred in 5 cases. Conclusion Laparoscopic total gastrectomy for gastric cancer is safe,feasible,less traumatic and of fast postoperative recovery.

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.

The operation path, lymph node dissection and reconstruction of the alimentary tract are the three most technical difficulties of laparoscopy-assisted gastrectomy. The essential difference between laparoscopy-assisted gastrectomy and open gastrectomy is the operation path. Based on our clinical experience, we investigated reasonable paths for laparoscopyassisted gastrectomy. Patients were placed in a supine position with their legs apart, and the operator stood on the left side of the patient. Five trocars were placed in the abdominal wall in a curved line. The operation was carried out in the order of greater gastric curvature, the lower region of the pylorus and antrum,the upper region of the pancreas, omentum minus, cardia, and arcuate diaphragm. From May 2004 to April 2010, we successfully carried out 761 laparoscopy-assisted gastrectomies with satisfactory outcomes.

Objective To explore the efficacy of laparoscopy-assisted radical gastrectomy for patients with gastric cancer. Methods The clinical data of 726 patients who received laparoscopy-assisted radical gastrectomy for gastric cancer at the Southwest Hospital from January 2004 to April 2010 were retrospectively analyzed. The operation time, operative blood loss, number of lymph nodes dissected, length of hospital stay and mobidity were evaluated using t test or chi-square test. The survival of the patients were evaluated by Kaplan-Meier method.Results Laparoscopy-assisted radical gastrectomy was successfully carried out on 707 patients, and 19 patients were converted to open surgery. The mean operation time, operative blood loss, number of lymph nodes dissected were (179 ±52)minutes, (87 ±51) ml and 33 ± 14, respectively. The average distances of proximal and distal resection margin to the tumors were (6.3 ± 1.9)cm and (5.6 ± 1.7)cm, respectively. The average time to flatus, time to fluid diet and length of hospital stay were (2.9 ± 1.4) days, (3.1 ± 1.7) days and (7.9 ± 3.5) days,respectively. The peri- and postoperative mobidities were 2.2% (16/726) and 4.0% (29/726), respectively. A total of 685 patients were followed up for 6-82 months (mean, 48.3 months), and the 5-year survival rate was 58.4%. Conclusions Laparoscopy-assisted radical gastrectomy is a feasible procedure with minimal trauma, low morbidity and quick recovery of patients.