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BACKGROUND: There are two minimally invasive ways of treating cholecystolithiasis combined with choledocholithiasis, but there remains some controversy regarding which technique is better, since they both have advantages and disadvantages. The one-step method involves laparoscopic cholecystectomy, laparoscopic common bile duct exploration, and primary close (LC + LCBDE + PC), while the two-step procedure consists of endoscopic retrograde cholangiopancreatography, endoscopic sphincterotomy, and laparoscopic cholecystectomy (ERCP + EST + LC). OBJECTIVE: This multicenter retrospective study aimed to analyze and compare the effects of the two techniques. METHODS: The data of patients who underwent either one-step LCBDE + LC + PC or two-step ERCP + EST + LC treatment for gallstones in the gallbladder and bile duct at the Shanghai Tenth People’s Hospital, Shanghai Tongren Hospital, and Taizhou Fourth People’s Hospital between January 1, 2015 and December 31, 2019 were collected, and the preoperative indicators of the two groups were compared. RESULTS: The surgical success rate of the one-step laparoscopic group was 96.23% (664/690), the transit abdominal opening rate was 2.03% (14/690), and there were 21 cases of postoperative bile leakage. The success rate of the two-step endolaparoscopic surgery was 78.95% (225/285), the transit opening rate was 2.46% (7/285), and there were 43 postoperative cases of pancreatitis and five of cholangitis. Postoperative cholangitis, pancreatitis, postoperative stone recurrence, postoperative hospitalization, and treatment costs were significantly lower (P< 0.05) in the one-step laparoscopic group than in the two-step endolaparoscopic group. However, the amount of intraoperative bleeding, the postoperative extraction time of the abdominal drainage tube, and the incidence of bile leakage were higher (P< 0.05) in the one-step laparoscopic group than in the two-step endolaparoscopic group. CONCLUSION: The two methods of treating choledocholithiasis combined with choledocholithiasis that were analyzed in this study were safe and effective, and each method had its own advantages.

Acute appendicitis is one of the most common causes of acute abdominal pain. Accurate diagnosis is often hindered due to various presentations that differ from the typical signs of appendicitis, especially the position of the appendix. A delay in diagnosis or treatment may result in increased risks of complications, such as perforation, which is associated with increased morbidity and mortality rates. Necrotizing fasciitis caused by perforated appendicitis is extremely rare. We herein report a case of 50-year-old man presenting with an appendiceal abscess in local hospital. After ten days of conservative treatment with intravenous antibiotics, the patient complained about pain and swelling of the right lower limb and computed tomography (CT) demonstrated a perforated appendix and gas and fluid collection extending from his retroperitoneal cavity to the subcutaneous layer of his right loin and right lower limb. He was transferred to our hospital and was diagnosed with necrotizing fasciitis caused by perforated appendicitis. Emergency surgery including surgical debridement and appendectomy was performed. However, the patient died of severe sepsis and multiple organ failure two days after the operation. This case represents an unusual complication of a common disease and we should bear in mind that retroperitoneal inflammation and/or abscesses may cause necrotizing fasciitis through lumbar triangles.

Circulating hepatocellular carcinoma cells may be detected by reverse transcription-polymerase chain reaction. We investigated the relationship between circulating hepatocellular carcinoma cells and hepatoma patient survival after different managements and survival periods.Peripheral vein blood (5 ml) samples were obtained from 113 patients with hepatocellular carcinoma and from 33 control subjects (9 with liver cirrhosis after hepatitis B, 14 with chronic hepatitis B, 10 healthy individuals) between January 1, 2009, and December 31, 2013. To detect circulating hepatocellular carcinoma cells in peripheral blood, alpha-fetoprotein messenger RNA was amplified from total RNA extracted from whole blood by reverse transcription-polymerase chain reaction.Alpha-fetoprotein messenger RNA was detected in 59 blood samples from the hepatocellular carcinoma patients (59/113, 52.2%). In contrast, there were no clinical control subjects whose samples showed detectable alpha-fetoprotein messenger RNA. The presence of alpha-fetoprotein messenger RNA in blood seemed to be correlated with the stage (by TNM classification) of hepatocellular carcinoma, serum alpha-fetoprotein value, and the presence of intrahepatic metastasis, portal vein thrombosis, tumor diameter and/or distant metastasis. In addition, alpha-fetoprotein messenger RNA was detected in the blood of 25 patients showing distant metastasis at extrahepatic organs (100%), in contrast to 32 of 88 cases without metastasis (36.4%). All the patients with hepatocellular carcinoma were followed. Seventeen patients with resection of a T 2 stage hepatocellular carcinoma had a survival of 3.2 years after surgical management, 38 cases with resection of a T3 stage hepatocellular carcinoma had a 1.3-year survival, and only 37 cases with T4 stage disease after different treatments except surgery survived for 0.6 years (P0.01).The presence of alpha-fetoprotein messenger RNA in peripheral blood may be an indicator of circulating hepatocellular carcinoma cells, which might predict hematogenous spreading metastasis in patients with hepatocellular carcinoma and may be a poor prognostic factor for hepatocellular carcinoma patients.

Background Gastric cancer is the second most common cause of death from cancer worldwide. Epidemiologic studies have examined the possible association between tomato products consumption and gastric cancer, but the relationship between tomato products and the risk of gastric cancer is controversial. We performed a meta-analysis of cohort and case–control studies to analyze this association. Methods We systematically searched MEDLINE and EMBASE and contacted authors to identify potential studies published from January 1966 to June 2012. We pooled the relative risks from individual studies using a random-effects model and performed heterogeneity and publication bias analyses. Results Twenty-one studies were eligible for our inclusion criteria, in a pooled analysis of all studies, consumption of large amounts of tomato products (in a comparison of the highest and lowest consumption groups) reduced the risk for gastric cancer (odds ratio, 0.73; 95% confidence interval, 0.60–0.90). The pooled OR of lycopene consumption and serum lycopene was 0.88 (95% CI = 0.67–1.16) and 0.79 (95% CI = 0.59–1.07), respectively. Conclusions Consumption of large amounts of tomato products is associated with a reduced risk of gastric cancer. However, because of potential confounding factors and exposure misclassification, further studies are required to establish these findings.

To study the effect of triple organ transplantation (liver, kidney, and pancreas) in patient of end-stage liver disease with renal failure and diabetes, and to explore the optimal surgical procedure.Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation was performed on a 43-year-old male patient with exocrine pancreatic insufficiency and insulin-dependent diabetes related to chronic pancreatitis (CP) who developed hepatic and renal failure. The pancreatic exocrine secretions were drained enterically to the jejunum. Prednisone, tacrolimus, mycophenolate mofetil, and ATG were used as immunosuppression therapy.Good liver and pancreas allograft function recovery was achieved within 7 days after the operation. And the recovery of renal allograft function was delayed. The renal allograft was removed because of break-down of renal blood flow 16 days after the transplantation. A new renal transplantation was performed at the same position. The second kidney graft recovered its normal function 3 days later. Up to the writing of this paper no acute rejection of organs and such complications as pancreatitis, thrombosis, and localized infection occurred. The patient became insulin independent with normal liver and renal function.Simultaneous piggyback orthotopic heterotopic liver, pancreas-duodenum, and kidney transplantation can be a good method for the patients with exocrine pancreatic insufficiency and insulin-dependent diabetes combined with hepatic and renal failure.

Expression of Fas ligand (FasL) on the graft by gene transduction is expected to introduce apoptosis to lymphocytes to protect rejection, but the FasL-expressing graft cells may also induce apoptosis as the graft usually expresses Fas antigens. In this study, a strong antiapoptotic gene, bcl-2, was cotransfected with the FasL gene in rat liver graft to protect against Fas-mediated cell death and to prolong recipient survival.Orthotopic liver transplantation was done in a strain combination of DA to LEW rats. After donor vascular isolation, adenovirus-mediated FasL and bcl-2 genes were cotransfected in the liver graft.Intragraft expression of FasL mRNA was constitutively expressed after adenovirus-mediated transduction, although expression of FasL increased mildly in control grafts. Bcl-2 mRNA was highly expressed at 2 days after reperfusion. In contrast, lower expression of bcl-2 was observed in the control group. The average survival of the gene transferred allografts increased from (9.8+1.3) days to (18.5+8.7) days compared with the control group.Our results indicate that rat liver allografts can be protected against host immune responses by adenovirus-mediated FasL and bcl-2 transfection, and that bcl-2 expression prevents the graft from Fas-mediated apoptosis.

To investigate the mechanism of enhancement of invasion of hepatocellular carcinoma (HCC) cells by lysophosphatidic acid (LPA).Human HCC cells of the line SMMC7721 were cultured. LPA at different concentrations (2, 5, and 25 micromol/L) was added into the culture fluid. The Rho activity was detected with Rho activity detection kit. Western blotting was used to detect the protein expression of Rho. Adhesion test was conducted to calculate the adhesion percentage of the SMMC7721 cells. The invasion potential of the SMCC7721 cells was observed using transwell cell test.LPA at the concentrations of 5, and 25 micromol/L increases the activity of Rho protein. When the concentration of LPA was 25 micromol/L the activity of Rho protein was 400 times that of the control group (P0.01). The Rho protein expression in the SMCC7721 cells increased when stimulated by LPA, peaked 20 approximately 25 hours after stimulation, and then gradually decreased. When the concentration of LPA was 25 micromol/L the Rho protein expression level was 242% higher than that of the control group. LPA at the concentration of 5 micromol/L and over increased the migratory and invading potential of the SMCC7721 cells and increased the adhesiveness of the SMCC7721 cells time-dependently.LPA increases the migratory and invading potential of HCC cells through Rho signal transduction pathway.

To obtain recombinant rat betacellulin with biological activity.A 534 bp of rat betacellulin gene fragment was amplified by RT-PCR from rat kidney and cloned into pET28a(+) vector to construct recombinant plasmid pET28a-rBTC. The recombinant plasmid was transformed into E. coli BL-21(DE3) and the betacellulin was expressed under IPTG induction. The expressed betacellulin was detected by SDS-PAGE and Western blot. The expressed protein was purified by Ni(2+) affinity chromatography and then renatured by dialysis. The effect of the renatured protein on proliferation of NIH3T3 cells was detected by MTT colorimetry.Rat betacellulin protein with M(r) being 20 000 was expressed under IPTG induction. The purity of purified protein reached over 96%. After renaturation, the expressed protein could significantly stimulate the proliferation of NIH3T3 cells.Rat betacellulin gene is successfully cloned into the expression vector pET28a(+) and highly expressed in E.coli. Purified and refolded betacellulin protein can obviously stimulate the proliferation of NIH3T3 cells.

To investigate the expression of human telomerase reverse transcriptase(hTERT) gene and its relationship with proliferating cell nuclear antigen and P53.Immunohistochemical staining was used to detect the expressions of hTERT, PCNA and P53 in 42 hepatocellular carcinoma(HCC) tissues. The relationship between hTERT expression, pathological characters of HCC and PCNA and P53 expression was analyzed.The expression rates of hTERT, PCNA and P53 in HCC tissues were 71.4%(30/42),76.2%(32/42) and 73.8%(31/42), respectively. hTERT was not expressed in normal liver tissues. The expression rates of hTERT gene in HCC tissues of different pathological grades (I, II and III)were 40.0%(4/10), 70.0%(14/20) and 100%(12/12), respectively. The expression of hTERT was correlated with HCC recurrence.The expression of hTERT gene may relate to the genesis and progression of HCC. There is no significant correlation between the expression of hTERT and PCNA and P53. The detection of hTERT gene expression may be regarded as a marker for recurrence of HCC.

To obtain massive human pancreatic islets with modified techniques and evaluation of the islets for the clinical allo-transplantation to treat type I and II diabetes.28 consecutive adult human pancreata were isolated with modified automated techniques. Islets were purified using continuous density gradient. The islet yield was counted with international standard known as islet equivalent (IEQ). The function of the isolated islets was evaluated by measuring DNA/insulin ratio, static glucose stimulating test in vitro and transplanting the islets into diabetic nude mice in vivo followed by abdominal glucose tolerance test and C peptide measurement.The yield of 28 consecutive human pancreata isolations ranged from 5 000 to 1 030 000 IEQs/pancreas with the average of 291 635 IEQs/pancreas. The first 13 isolations yielded 49 123 IEQs/pancreas, 846 IEQs/g and, purity 87% in average. The remained 15 isolations after the modifications yielded 501 813 IEQs/pancreas, 7 003 IEQs/g and purity 89% in average. The results of in vitro SGS showed good response to the different glucose concentration. 34 diabetic nude mice were transplanted under the renal capsule with the freshly isolated islets. 29 out of 34 diabetic mice obtained normoglycemia within 12 hours and the glucose tolerance tests were near normal. Serum C peptide level of transplanted mice is close to that of the control group.Massive human islets can be isolated with the modified techniques. Quality assessment of these islets both in vitro and in vivo has indicated that these high quality human islets could be used for the clinical allogeneic islet transplantation.

AIM: To investigate the effects of proliferation and apoptosis induced by cyclooxygenase-2 (COX-2) inhibitor celecoxib in combination with cisplatin. METHODS: The human pancreatic cancer cell line BxPC-3 cells were treated with COX-2 inhibitors celecoxib and cisplatin. The cell relative viability was examined using 3 (4, 5-dimethylethiazoly 1-2-) 2, 5-diphonyl tetrazolium bromide (MTT) assays. the expression of COX-2 mRNA was detected by RT-PCR, flow cytometry and Hoechst-33258 were used to demonstrate apoptotic changes in celecoxib and cisplatin treated cells. RESULTS: After treatment of BxPC-3 cells with celecoxib, as measured by MTT, cell viability was inhibited in a dosedependent and time-dependent manner with an IC50 of 100 nM at the time of 24h. The expression of COX-2 mRNA could be significantly decreased by celecoxib. Furthermore, we demonstrated that the combination of celecoxib with cisplatin inhibited cell growth and induced cell apoptosis to a greater degree than either compound alone. The apoptotic morphologies were demonstrated by Hoechst-33 258. CONCLUSION: Combination of celecoxib with cisplatin inhibits cell proliferation and induces cell apoptosis, and the potent effectiveness of celecoxib in combination with gemcitabine may hold a promise in the clinical treatment of pancreatic cancer. Liu JW, Li KZ, Dou KF, Song ZS, Su MQ, Yu WB. Effects of COX-2 inhibitor with cisplatin on proliferation and apoptosis of pancreatic cancer cells. Shijie Huaren Xiaohua Zazhi 2004;12(5):1139-1143

Background MicroRNAs are a class of small non-coding RNAs that play an important role in various human tumor initiation and progression by regulating gene expression negatively. The aim of this study was to investigate the effect of miR-214 on cell proliferation, migration and invasion, as well as the functional connection between miR-214 and PTEN in gastric cancer. Methods miR-214 and PTEN expression was determined in gastric cancer and matched normal tissues, and human gastric cancer cell lines by quantitative real-time PCR. The roles of miR-214 in cell proliferation, migration and invasion were analyzed with anti-miR-214 transfected cells. In addition, the regulation of PTEN by miR-214 was evaluated by Western blotting and luciferase reporter assays. Results miR-214 was noted to be highly overexpressed in gastric cancer tissues and cell lines using qRT-PCR. The expression level of miR-214 is significantly associated with clinical progression and poor prognosis according to the analysis of the clinicopathologic data. We also found that the miR-214 levels are inversely correlated with PTEN in tumor tissues. And PTEN expression level is also associated with metastasis and invasion of gastric cancer. In addition, knockdown of miR-214 could significantly inhibit proliferation, migration and invasion of gastric cancer cells. Moreover, we demonstrate that PTEN is regulated negatively by miR-214 through a miR-214 binding site within the 3’-UTR of PTEN at the posttranscriptional level in gastric cancer cells. Conclusions These findings indicated that miR-214 regulated the proliferation, migration and invasion by targeting PTEN post-transcriptionally in gastric cancer. It may be a novel potential therapeutic agent for gastric cancer.