Objective:To explore the clinical features of recurrence after choledocholithotomy and to analyze the risk factors.Methods:The clinical data of 730 patients with choledocholithiasis who were treated in our hospital from January 2005 to July 2016 were analyzed retrospectively,550 cases who were received choledocholithotomy were defined as laparotomy group,30 cases with laparoscopic common bile duct exploration (LCBDE) were defined as the LCBDE group,and 150 cases with endoscopic sphincterotomy (EST) were defined as EST group.The recurrence rate of the three groups were compared.The patients of three groups were divided into recurrence group (n=227) and non recurrence group (n=503) according to the recurrent situation,then the clinical features and risk factors of recurrent patients were analyzed by univariate and multivariate Logistic regression analysis.Results:The recurrence rate of EST group was 38.67％,which was significantly higher than that of LCBDE group with 26.67％ and the laparotomy group with 29.27％,and there was statistical difference (P＜0.05).The results of univariate analysis showed that there were statistically significant differences in age,history of HBV infection,jaundice,abnormal total bilirubin,peripapillary diverticulum,biliary infection,biliary stricture,papillary stenosis,sphincter of Oddis dysfunction,history of biliary surgery,cholecystectomy,bile duct diameter ≥ 15 mm,bile duct angle ≤120°,operation type,stone quantity ≥ 2 grains,stone diameter ≥ 10 mm,with or without gallstones (P＜0.05).The results of Logistic multivariate regression analysis showed that age,having peripapillary diverticulum,having history of biliary surgery,bile duct diameter ≥ 15 mm,stone quantity ≥ 2grains and EST operation type were the independent risk factors of the recurrence after choledocholithotomy (P＜0.05).Conclusion:There are many risk factors of recurrence after choledocholithotomy,and operation method should be based on the size and the number of the stones,and the constitution of patients.Preventive measures should be strengthened to control the recurrence after choledocholithotomy.

As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cricetinae ; Cricetulus ; Gene Expression Profiling ; Green Fluorescent Proteins ; genetics ; Humans ; Molecular Sequence Data ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Tissue Distribution ; Transfection

Translational medicine is a systemic project because it is patient and clinical problems oriented, aiming at research results application, and involves multidisciplinary cooperation. Studies on molecular events in the precancerous stage, early stage and metastasis of colorectal cancer (CRC) are the CRC hot research topics currently. Investigations on the earliest molecular events can help to find out the markers which may improve the effect of CRC screening and predict CRC liver metastasis and prognosis. Based on the concept of micro environment, molecular targeted drugs to interfere with metastasis and invasion and new concepts of surgical resection margin and neoadjuvant therapy will gain recognition from clinicians.
Colorectal Neoplasms ; Humans ; Translational Medical Research

Objective The aim of the present study was to investigate the relationship between the intake of salt and salted food and the infection of Helicobacter pylori (Hp) among 40-69 years old local residents in a county with high gastric cancer risk in Anhui province. Methods From July 2015 to August 2018, we conducted a questionnaire and a serological test for Hp among 40-69 years old local residents in Lujiang county, Anhui province. The questionnaire focused on the consumptions of salt and salted food. The relationship between Hp infection and risk factors was analyzed by gender. Univariate and multivariate logistic regression analysis were used to analyze the relevant influencing factors. Results The Hp infection rate of total local residents was 50.07%. Among male subjects, age, body mass index(BMI), marital status, educational level, job, labor intensity and income had no link to Hp infection (all P>0.05). But among female subjects, BMI was associated with Hp infection ( 2=13.454,P=0.001). Besides, alcohol consumption was a risk factor for Hp infection in male subjects(OR=1.789，95% CI:1.188-2.694，P=0.003). But, high intake of salt and salted food had no effect on Hp infection after adjustment for alcohol consumption variable in men using multivariate analysis (all P>0.05). After adjusted for BMI variable among female individuals, high salt intake (≥9 g/day) (OR=1.462，95% CI:1.060-2.015，P=0.021) and the high salted food intake (≥1 times /day) were risk factors for Hp infection in women(OR=1.560，95% CI:1.021-2.383，P=0.040). Conclusions In one county with high gastric cancer risk in Anhui province, high salt intake (≥9 g/day) and high salted food intake (≥1 times/day) are risk factors for Hp infection among 40-69 years old female local residents.

OBJECTIVETo study an intervention model of "schools without infected students with schistosoma japonica", to control and prevent students from schistosoma infection.

METHODSTwelve primary schools of four heavy endemic counties (districts) with schistosomiasis in the Poyang Lake areas were selected as the study fields, of which, ten schools were the experimental groups, and the other two schools were the control groups by cluster random sampling. All enrolment students were the target population. The baseline survey was carried out in 2005, and an intervention model, "information dissemination + behavior participation + behavior encouragement", was applied in the experiment groups in 2006 - 2008, then the effect of intervention was assessed.

RESULTSBefore intervention (2005), the anti-schistosomiasis knowledge awareness rate of experimental and control groups were 14.75% (324/2196) and 16.58% (91/549), and the different was not significant (χ(2) = 1.14, P > 0.05); the rate of accurate attitude of anti-schistosomiasis were 14.71% (323/2196) and 11.84% (65/549) in experimental and control groups, and the difference was not significant (χ(2) = 2.98, P > 0.05); the rate of contacting infected water were 15.44% (18 988/122 976) and 15.03% (4622/30 744) in experimental and control group and the difference was not significant (χ(2) = 3.13, P > 0.05); and the infection rate of schistosomiasis of experiment control groups were 9.65% (212/2196) and 10.56% (58/549), the difference was not significant (χ(2) = 0.41, P > 0.05). After one year intervention (2006), the anti-schistosomiasis knowledge awareness rate of experimental and control groups were 97.79% (2032/2078) and 18.11% (98/541), and the different was significant (χ(2) = 1794.31, P < 0.01); the rate of accurate attitude of anti-schistosomiasis were 99.09% (2059/2078) and 13.49% (73/541) in experimental and control group, and the difference was significant (χ(2) = 2077.45, P < 0.01). After 1 - 3 years intervention (2006 - 2008), there were no any contactors with infected water and infectors with schistosome in students of the experiment group in successive 3 years. While in the control group of the same period, the rate contacting infected water were 16.12% (4884/30 296), 11.11% (3079/27 720) and 12.25% (3451/28 168); the infection rate of schistosomiasis were 8.87% (48/541), 7.47% (37/495) and 7.95% (40/503), respectively.

CONCLUSIONThe intervention model of health promotion, "information dissemination + behavior participation + behavior encouragement", can effectively control and prevent students from infecting schistosoma japonica in heavy endemic areas with schistosomiasis.

Animals ; Health Promotion ; Humans ; Schistosomiasis ; prevention & control ; Schistosomiasis japonica ; School Health Services ; Schools ; Students

AIMBLT1 and BLT2 were both recently cloned and identified as two subtypes of leukotrine B4 (LTB4) receptors. With the usage of U-75302 and LY255283, the specific antagonists of BLT1 and BLT2 respectively, the involvement of BLT1 and BLT2 in the inflammatory and immunological responses was in vitro explored.

METHODS(1) To investigate inhibition of U-75302 and LY255283 on the proliferation of rat synovial cells, 3H-TdR incorporation into the cells was quantified. (2) Flow cytometric assay for interferon-gamma (IFN-gamma) and interleukine 4 (IL-4) profiles in CD4+ T lymphocytes from rat spleen was carried out to determine the ratio of Th1/Th2.

RESULTS(1) For inhibition on rat synovial cells proliferation, U-75302 exerted its effect only at a high concentration of 10 micromol/L and LY255283 at the concentrations of 10 micromol/L-10 micromol/L. (2) Both U-75302 and LY255283 could elevate the percentage of Th2, but could not influence that of Th1.

CONCLUSIONBLT1 and BLT2 were involved in the synovial cells proliferation change the ratio of Th1/Th2. Their meaning served as targets for prevention and treatment of infectious diseases should be emphasized.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Fatty Alcohols ; pharmacology ; Glycols ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Wistar ; Receptors, Leukotriene B4 ; antagonists & inhibitors ; physiology ; Synovial Membrane ; cytology ; immunology ; Tetrazoles ; pharmacology ; Th1-Th2 Balance

OBJECTIVETo investigate the effects and complications of primary and secondary placements of motility coupling post (MCP) in the unwrapped porous polyethylene orbital implant (PPOI) following enucleation.

METHODSWe investigated 198 patients who received PPOI implantation following the standard enucleation procedure in the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China, from 2002 to 2004. These patients were subgrouped into PPOI-only patients (112 cases, received PPOI following enucleation), primary MCP patients (46 cases, received primary placement of MCP during PPOI operation), and secondary MCP patients (40 cases, received secondary placement of MCP 6 months after the initial surgery). Effects and complications among these three groups were compared.

RESULTSThe PPOI-only patients took shorter treatment course when compared with other two MCP groups (P<0.001), without significant difference noted between the two MCP groups. However, the two MCP groups had better prosthetic motility than PPOI-only group (P<0.001), without significant difference between the two MCP groups. In the early stage, 2 eyes in the PPOI-only group and 1 eye in the primary MCP group had PPOI infection. In PPOI-only group, 3 (2.68%) eyes had PPOI exposure, which occurred after fitting the prostheses; 4 eyes (8.70%) in primary MCP group and 1 eye (2.50%) in secondary MCP had PPOI exposure, which occurred before fitting the prostheses. After prosthesis was fit successfully, the excessive discharge and granuloma were 33.9% and 1.79% in PPOI group-only, 53.3% and 8.9% in primary MCP group, and 52.5% and 7.5% in secondary MCP group, respectively.

CONCLUSIONBoth primary and secondary placements of MCP into the PPOI following enucleation can help patients to obtain desirable prosthetic motility, but may be associated with more complications. The primary placement of MCP with skilled operation in selected patients is more recommendable than secondary placement.

Adult ; Biocompatible Materials ; Eye, Artificial ; adverse effects ; Female ; Granuloma ; etiology ; Humans ; Infection ; etiology ; Male ; Middle Aged ; Movement ; Orbital Implants ; adverse effects ; Polyethylene ; Postoperative Complications ; etiology

OBJECTIVETo establish a simple, rapid and economical method in detecting mutations of oncogene K-ras and to investigate its mutations in colorectal cancer tissues and its relationship with clinicopathologic characteristics of colorectal carcinoma.

METHODSForty colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using polymerase chain reaction (PCR) followed by direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by sequence analysis. The other 113 colorectal cancer tissues were tested for K-ras mutations at codon 12 and codon 13 using PCR-RFLP followed by sequence analysis only. The mutation results were analyzed with the corresponding clinical pathological data.

RESULTSAmong 40 colorectal cancer cases, none of K-ras mutations at codon 12 and codon 13 was detected by PCR followed by direct sequencing. However, K-ras mutations were found in 11 cases (11/40, 27.5%) by PCR-RFLP followed by sequence analysis, including 8 cases at codon 12 and 3 cases at codon 13 respectively. Among 153 colorectal cancer cases, point mutations were detected by PCR-RFLP followed by sequence analysis in 58 cases (37.9%). Point mutations at codon 12 were found in 46 cases and 12 cases at codon 13. Mutations with the highest frequency were G→A transitions (25/58, 43.1%) at codon 12. No significant correlation was observed between mutations of K-ras and gender, invasive depth, tumor differentiation, number of invaded lymph nodes, distant metastasis and clinical stage (P > 0.05). Mutation of oncogene K-ras at codon 12 and codon 13 was closely related with age and tumor location (P < 0.05). The incidence of K-ras mutation was significantly higher in younger patients and in patients with ascending colon cancer.

CONCLUSIONSPCR-RFLP followed by sequence analysis is a rapid, simple, sensitive and low-cost method. It is a suitable technology for detecting hot-spot mutations in the K-ras oncogene. Mutation of oncogene K-ras at codon 12 and codon 13 is a common molecular event in colorectal carcinogenesis, which might be related with age and tumor location.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Polymorphism, Restriction Fragment Length

OBJECTIVETo cooperate with the study of HBV vector, hygromycin-resistant packaging cell line was developed that allows encapsidation of plasmids into HBV particles.

METHODSFree of packaging signal, HBV genome was inserted into plasmid pMEP4, which expresses the HBV structural proteins including core, pol and preS/S proteins. HepG2 cell lines were employed to transfect with the construct. Hygromycin selection was done at a concentration of 150 micrograms/ml in the culture medium. The hygromycin-resistant clones with the best expressions of HBsAg and HBcAg were theoretically considered as packaging cell line and propagated under the same conditions. It was infected with recombinant retrovirus vector and hen selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSHygromycin-resistant HBV packaging cell line was generated, which harbored an HBV mutant whose packaging signal had been deleted. Expressions of HBsAg and HBcAg were detectable. Infected with recombinant retrovirus pRV-CP, the hygromycin-resistant packaging cell line was found to secrete mutant HBV particles and no wild-type HBV was detectable in the culture medium.

CONCLUSIONAfter the packaging signal was deleted and transfected into HepG2 cell lines, the partial HBV genome lost its ability to form wild-type HBV, but conserves cis-action providing structural proteins for the packaging of the replication-defective HBV.

Cell Line ; Drug Resistance, Viral ; Genetic Vectors ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Hygromycin B ; pharmacology ; Mutation ; Plasmids ; Retroviridae ; genetics ; Transfection ; Virus Assembly

BACKGROUNDTo explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular expression of dominant negative mutants of core protein.

METHODSTwo kinds of full length mutant HBV genome, which express Core-partial P and Core-S fusion protein, were transfected into HepG 2.2.15 cell lines. Positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA and intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR.

RESULTSThe mean inhibitory rates of HBsAg were 2.74+/-3.83%, 40.08+/-2.05% (P less than 0.01) and 52.94+/-1.93% (P less than 0.01) in group 2.2.15-pMEP4, 2.2.15-CP and 2.2.15-CS, respectively. The mean inhibitory rates of HBeAg were 4.46+/-4.25%, 52.86+/-1.32% (P less than 0.01) and 41.60+/-1.65% (P less than 0.01), respectively. The inhibitory rates of HBc related HBV DNA were 15.3%, 82.0% and 67.2%, respectively. Recombinant HBV virion was detectable in the culture medium of only group 2.2.15-CP.

CONCLUSIONDominant negative mutants of core protein can efficiently suppress wt-HBV replication and the expressions of HBV antigens. With the help of wild-type HBV, the recombinant HBV genome can form and secret HBV like particles, which provides evidence that the antiviral gene will be hepatotropic expression and the antiviral effects will be amplified.

Cell Line, Tumor ; Genetic Engineering ; Genetic Therapy ; Genetic Vectors ; Genome, Viral ; Hepatitis B Core Antigens ; biosynthesis ; physiology ; Hepatitis B virus ; genetics ; Humans ; Point Mutation ; Recombinant Fusion Proteins ; biosynthesis ; physiology ; Virus Replication