Objective To study the frequency of peripheral blood T helper 17(Th17)and regulatory T cells(Treg)and their as-sociated cytokines level in child chronic hepatitis B(CHB)and to investigate the balance characteristics of Th1 7/Treg under differ-ent progression status in child CHB.Methods Peripheral blood and serum of 60 children with CHB and 16 healthy controls were collected for detecting the frequency of peripheral blood Fox3+Treg and Th1 7 by the flow cytometry and serum levels of Th1 7/Treg differentiation associated cytokine IL-17A,TGF-β,IL-22,IL-23,IL-6 and IL-1βwere detected by the enzyme-linked immu-nosorbent assay(ELISA).Results The frequency of peripheral blood Th1 7 and Treg,and the ratio of Th1 7/Treg in the CHB group were significantly higher than those in the healthy controls,the frequency of Th1 7 was positively correlated with disease progres-sion.In addition,serum IL-17A,IL-22 and IL-23 levels in the CHB children were higher than those in the healthy controls(P<0. 05).Conclusion Th17 and Treg play the important role in the mechanism of child CHB.The imbalance of Th17/Treg exists in child CHB.

Objective To investigate the effects of laparoscopic operation in treating type Ⅲ child biliary atresia and its influence on postoperative infections .Methods 85 children cases of type Ⅲ biliary atresia were randomly divided into the conventional sur-gery group(OP group ,n=42) and the laparoscopic surgery group(LP group ,n=43) .The operation situation ,postoperative changes of inflammatory factors ,changes of liver function indexes before and after operation and postoperative infection were compared be-tween the two groups .Results The operative time of the LP group was longer than that of the OP group (P<0 .05) ,the intraoper-ative blood loss and the VAS scores of the LP group were less than those of OP group (P<0 .05) ,the postoperative eating recovery time of the LP group was earlier than that of the OP group (P<0 .05) ,the length of hospital stay of the LP group was shorter than that of the OP group(P<0 .05) and the postoperative infection rate of the LP group was lower than that of the OP group (P<0 . 05) .The liver function indexes recovery showed no statistically significant differences before and in postoperative 1 ,3 ,6 ,12 months between the two groups(P>0 .05) .Conclusion The laparoscopic operation and the conventional operation in treating type Ⅲ child biliary atresia have their own advantages and disadvantages .The appropriate operation mode should be selected according to the children′s actual situation .

Objective To investigate the effects of celastrol on the cell growth and apoptosis of human gallbladder cancer NOZ cells,and explore its potential molecular mechanism.Methods NOZ cell were cultured in vitro.And CCK-8 assay,Annexin V-FITC/PI staining method,cell cycle analysis were conducted to investigate the effects of celastrol on the growth and apoptosis of NOZ cells after being treated with drugs.The mitochondrial membrane potential and Bax and Bcl-2 protein expression level were determined by Rhodamine 123 and Western blot,respectively.Results Celastrol could inhibit NOZ cell growth,and the IC50 value was 5.3 μmol/L.Annexin-V/PI staining showed that cell apoptosis of NOZ cells were induced as the celastrol concentration increased,and the apoptosis ratio of control group was 4.4％,while the apoptosis rates of the test groups (2,5,10 p mol/L) were 7.4％,27.1％ and 43.4％,respectively.In addition,cell cycle analysis revealed that celastrol could induce G1-phase arrest.The G1-phase rate of control group was 25.6％,while the G1-phase rates of the test groups (2,5,10 μmol/L) were 36.5％,45.7％ and 92.5％,respectively.The mitochondrial membrane potential was measured after treatment with celastrol and the results indicated that the mitochondrial membrane potential was significantly decreased.Western Blot showed that the protein expression of Bax increased and Bcl-2 decreased in a time-dependent manner after treatment with celastrol.Conclusions Celastrol may inhibit cell proliferation of human gallbladder cancer NOZ cells and induce cell apoptosis partly by inducing the loss of mitochondrial membrane potential.

AIM: To establish a sensitive and specific liquid chromatobraphy-mass spectrometry(time-of-flight)[LC-MS(TOF)] for the determination of astragaloside Ⅳ in rabbit plasma. METHODS: The HPLC-MS utilizing solid phase extraction was established to determine the concentration of astragaloside IV and ginsenoside R_~g1 , was used as internal standard. The analysis was carried on Agilent Hypersiol ODS(5 ?m, 4.6 mm?250 mm) column with a mobile phase of methanol-water (80∶20, v/v).Detection was performed on a time-of-flight mass spectrometry equipped with an ESI internal and operated in positive-ionization mode. Astragaloside Ⅳ quantitation was realized by computing the peak area ratio (astragaloside Ⅳ-ginsenoside R_~g1 )(astragaloside Ⅳ m/z807[M+Na]+ and ginsenoside R_~g1 m/z823[M+Na]+) and comparing them with calibration curve (r=~0.999 ). RESULTS: The linear calibration curve was obtained in the concentration range of 0.01-5 ?g?L~-1 .The detection limit of astragaloside Ⅳ was 0.01 ?g?L~-1 .The average recovery was more than 98%.The intra-and inter-run precision was measured to be below 5% of RSD. CONCLUSION: The method is sensitive, simple and rapid ,so, it can meet the need for the pharmacokinetics and bioavailability of astragaloside Ⅳ.

OBJECTIVETo investigate the risk factors of early surgical intervention in Crohn's disease (CD) patients with spontaneous intra-abdominal abscess.

METHODSClinical data of 94 CD patients with spontaneous intra-abdominal abscess admitted to The Sixth Affiliated Hospital of Sun Yat-sen University between May 2008 and Dec 2015 were analyzed retrospectively. Univariate and multivariate analysis were applied to evaluate the early surgery risk of CD patients with spontaneous intra-abdominal abscess using logistic regression model.

RESULTSA total of 94 eligible patients were identified from our registry, including 70 males and 24 females. The mean age at the diagnosis of CD and at development of abscess was 28.4 years and 30.4 years old, respectively. The median duration of CD between the diagnosis and development of an abscess was 3 years. According to the Montreal classification, L3 (ileocolonic) was the most common disease location (81.9%) in these patients. Most of the patients(76.6%) developed a single abscess, while multiple abscesses were detected in 22 patients(23.4%). Forty-four patients(46.8%) underwent surgery within 60 days after hospitalization due to spontaneous intra-abdominal abscess complicating CD. Multivariate logistic regression analysis revealed that history of abdominal surgery(OR=3.23, 95%CI:1.12 to 9.31, P=0.030), concomitant intestinal stenosis (OR=3.52, 95%CI:1.26 to 9.85, P=0.017) and concomitant intestinal fistula (OR=4.31, 95%CI:1.25 to 14.80, P=0.020) were the independent risk factors of early surgical intervention, while enteral nutrition (OR=0.18, 95%CI:0.05 to 0.62, P=0.007) was the independent protective factor.

CONCLUSIONSNearly half of CD patients with spontaneous intra-abdominal abscess will undergo early surgical intervention. Patients with history of abdominal surgery, concomitant intestinal stenosis and concomitant intestinal fistula have higher risk of early surgical intervention, and appropriate application of enteral nutrition may reduce the risk.

Objective:To evaluate extrathecal vs. intrathecal approach of Glisson's pedicle in laparoscopic hepatectomy of hepatocellular carcinoma (HCC). Methods:Clinical data of 96 HCC patients receiving laparoscopic hepatectomy at the Department of Hepatobiliary Surgery, the 900 Hospital of the United Logistics Force from Jan 2019 to Jun 2022 were retrospectively analyzed. Thrity-six cases were treated with extrathecal approach of Glisson's pedicle and 60 cases were by intrathecal approach.All those 60 undergoing intrathecal approach cases were matched by 1∶1 propensity score matching, and the caliper value was 0.02. The duration of surgery, blocking-time of hepatoduodenal ligament, intraoperative blood loss, indexes of the liver function and the prothrombin time on the 1st day of the postoperative period, rate of the postoperative complications, postoperative hospital stay, postoperative survival and recurrence were compared between the two groups.Results:After propensity score matching, 32 cases in each group were identified. The operation time of the extrathecal group was shorter than that of the intrathecal group [(207.5±77.4) minutes vs. (248.6±74.7) minutes, P=0.03], the intraoperative bleeding volume was less than that of the intrathecal group [(150.0(100.0, 300.0) ml vs. 250.0(150.0, 587.5) ml, P=0.01], and the albumin level was higher on the 1st day after the surgery [(36.5±3.2) g/L vs. (34.3±3.2) g/L, P<0.01]. There was no statistically significant difference in postoperative liver function, coagulation, complications, hospitalization time, all over survival rate between the two groups (all P>0.05). Conclusion:Extrathecal approach for Glisson's pedicle is safe and feasible in laparoscopic hepatectomy for HCC, with simpler surgical procedure, less intraoperative trauma.

OBJECTIVETo investigate the influence of CCL21 on the invasion and metastasis of colorectal cancer (CRC).

METHODSCCL21 over-expressing CRC cell line was constructed by lentivirus infection and CCL21 low-expressing CRC cell line was constructed by lipofection. The effects of CCL21 on the invasion and metastasis of CRC cells and the stem cell-like phenotype were investigated by Transwell migration, invasion assay, wound healing assay and sphere formation assay.

RESULTSReal-time quantitative PCR and western blot confirmed that the expression of CCL21 was up-regulated by lentiviral transfection and down-regulated by siRNA liposome transfection. In vitro, Transwell assays showed that the invasion and migration in CCL21 over-expressing CRC cells decreased significantly as compared to those of CCL21 low-expressing cells. In wound healing assay, the CCL21 over-expressing CRC cells showed a significantly lower rate of migration. In addition, the sphere formation rate and density of CCL21 over-expressing CRC cells were lower than those with low-expression of CCL21.

CONCLUSIONCCL21 can suppress the migration and invasion of CRC cells and weaken their stem cell-like phenotype.