AIM:To evaluate effects of different chemotherapeutic agents on reversing the acquired resistance to TRAIL gene and clarify the involved mechanisms in DLD1-TRAIL/R colon cancer cells.METHODS: Human colon cancer cell line DLD1-TRAIL/R cells that were resistant to TRAIL-expressing adenovector(Ad/gTRAIL) were treated with Ad/gTRAIL combined with different chemotherapeutic agents.Then,the cell viability was measured by MTT method,and apoptotic signaling conditions,including activation of caspase-3 and caspase-8,expression of Bax and Bcl-XL,were measured by Western blotting analysis.RESULTS: In vitro data showed that several chemotherapeutic agents,including 5-fluorouracil(5-FU) and mitomycin c(MMC),overcome the acquired resistance to TRAIL gene in DLD1-TRAIL/R colon cancer cells.The combination of Ad/gTRAIL and 5-FU effectively suppressed tumor growth in vivo in subcutaneous tumors established from DLD1-TRAIL/R cells.Further data showed that treatment with the combination of Ad/gTRAIL and 5-FU or MMC led to enhance the activation of caspase-3.Moreover,MMC but not 5-FU induced overexpression of Bax gene that was sufficient to overcome the resistance to TRAIL gene in DLD1-TRAIL/R cells.CONCLUSION: Chemotherapeutic agents,such as 5-FU and MMC,overcome the acquired resistance to TRAIL gene in DLD1-TRAIL/R cells.The candidate mechanisms for MMC but not 5-FU to overcome this resistance might involve the induction of over-expressed Bax protein in DLD1-TRAIL/R cells.

OBJECTIVETo evaluate the efficacy and safety of long-term on-demand use of vardenafil in the treatment of erectile dysfunction (ED).

METHODSWe conducted a questionnaire investigation among 891 ED patients treated by on-demand use of oral vardenafil at 20 mg every 3 days from March 2007 to January 2010, covering the general information of the patients, their need for and attitudes towards the treatment, clinical efficacy and adverse events of the drug, and satisfaction of the patients and their partners after 12 weeks of treatment.

RESULTSTreatment and follow-up were completed in 700 patients, of whom 504 (72%) achieved sufficient hardness and duration of penile erection and overall sexual satisfaction, 84 (12%) admitted to improvement of erectile hardness and duration but not adequate satisfaction, and the other 112 (16%) experienced no significant changes. Significant differences were found in IIEF-5 scores, Rigiscan test results and partners TSS scores before and after the treatment (P < 0.05). Most frequent adverse events included flushing (15%), dizziness and headache (10%), dyspepsia (3%), and nasal congestion (1%).

CONCLUSIONLong-term on-demand use of oral vardenafil, in addition to its safety and good tolerance, can effectively improve the erectile function of ED patients, their success rate of sexual intercourse, and overall quality of sexual life.

Adult ; Erectile Dysfunction ; drug therapy ; Follow-Up Studies ; Humans ; Imidazoles ; adverse effects ; therapeutic use ; Male ; Piperazines ; adverse effects ; therapeutic use ; Sulfones ; adverse effects ; therapeutic use ; Treatment Outcome ; Triazines ; adverse effects ; therapeutic use ; Vardenafil Dihydrochloride ; Vasodilator Agents ; adverse effects ; therapeutic use ; Young Adult

This study was aimed to compare the expression level of eIF4E in patients with leukemia and normal controls, and to explore its role in leukemogenesis. White blood cells were collected in 76 leukemia patients and 10 healthy volunteers. The mRNA and protein expressions of eIF4E were detected by QT-PCR and Western blot in 39 cases of acute myeloid leukemia (AML), 15 cases of chronic myeloid leukemia (CML), 22 cases of acute lymphocytic leukemia (ALL) and 10 healthy volunteers as normal controls. The results demonstrated that compared with normal controls, the absolute expression levels of eIF4E mRNA increased in patients with AML, ALL and CML in blastic phase (P < 0.05), but had no significant change between groups of CML in chronic and accelerated phase although some increasing in group of CML in accelerated phase. The relative expression level of eIF4E mRNA had no significant change in AML, ALL, CML groups except the two subtypes of leukemia M4 and M5. Furthermore, the protein expression level in group of CML in accelerated phase and blastic phase and all acute leukemia patients including AML and ALL were higher than that in normal controls (P < 0.05). It is concluded that although its mRNA relative expressions had no significant change in most leukemia patients, the absolute expression level of eIF4E mRNA and its protein expression is up-regulated in most leukemia patients, which may play an important role in leukemogenesis, so the eIF4E may be a promising target for leukemia therapy and eIF4E-targeted therapy may be an option especially for the relapse and refractory leukemia.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Eukaryotic Initiation Factor-4E ; genetics ; metabolism ; Female ; Humans ; Leukemia ; genetics ; metabolism ; pathology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Acute ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; RNA, Messenger ; genetics ; Young Adult

Objective To explore the relationship between alcohol drinking and fatty liver disease and its influencing factors.Methods From October to December,2013,a total of 394 people who underwent a physical examination in the medical examination center of Wenzhou central hospital were selected for this study.Anthroposomatology measurement and biochemical tests were conducted.Results There were significantly statistical differences of triglyceride,uric acid and cholesterol in different drinking groups (P <0.05).The prevalence of high triglyceride and uric acid were increased with alcohol consumption (P <0.05).There was no significant difference of alcohol consumption between fatty liver and non -fatty liver group (P =0.42).Logistic regression showed that waist -hip ratio,hypertension,overweight,obesity and hypertriglyceridemia were risk factors of fatty liver,while daily alcohol consumption cannot be thought as risk factor yet. Conclusion Waist -hip ratio,hypertension,overweight -obesity and hypertriglyceridemia were the risk factors of fatty liver.Alcohol consumption could contribute to the prevalence of triglyceride and uric acid.

OBJECTIVEEarly gestational mammalian fetuses possess the amazing ability to heal cutaneous wounds in a scarless fashion. Over the past years, scientists have been working to decipher the mechanisms underlying this regenerative repair. The remarkable phenotypic differences between fetal and adult healings behooves us to learn their characteristics in genetics, which represents potentially important mechanisms involved in wound repair observed in fetal versus adult tissues. In this sense, it is reasonable to construct subtractive cDNA library for future research.

METHODSMiddle laparotomy and hysterotomy were performed on pregnant rabbits at 20-day gestation to expose the fetal back, and a longitudinal incision through the skin was made on the back of the fetus. The traumatized fetal skin was harvested 12 hours post-operation, the fetus control and traumatized adult skin specimens were taken at the same time. dscDNA was synthesized from total RNA of skin samples with SMART technology. Taking one of the three samples as Tester respectively and the other two as Drivers, we obtained 1 forward and 2 reverse hybridization products. After being amplified with selective polymerase chain reaction, the products were inserted into a vector, and then transferred into E. coli HB101. The colonies were screened afterwards.

RESULTSThe wounded fetuses were alive for a long time even after birth. Every determinant step, such as RNA isolation, cDNA synthesis, Rsa I digestion, adaptor ligation and hybridization, was well-operated. Subtractive efficiency identification demonstrated that the suppression subtractive hybridization (SSH) was successful. Insertion into vector and transferring to E. coli were satisfactory.

CONCLUSIONSInstead of classic SSH, an improved SSH with 2 Drivers was applied for the experiment. Results confirmed that the improved program was reasonable and correct in both theory and practice. The subtractive cDNA library we have obtained is going to be used for future researches to reveal scarless healing related gene(s) and its (their) expression.

Animals ; Cicatrix ; genetics ; DNA, Complementary ; Disease Models, Animal ; Female ; Fetus ; Gene Library ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy, Animal ; RNA, Messenger ; analysis ; Rabbits ; Sensitivity and Specificity ; Skin ; injuries ; Wound Healing ; genetics ; physiology

OBJECTIVETo establish a RP-HPLC method for simultaneous determination of total quercetin, kaempferol and isorhamnetin in rat plasma after oral administration of Folium Mori extract (FME).

METHODSAfter a single dose of FME (110 mg/kg) was taken, rat plasma samples were collected. The samples were hydrolyzed with hydrochloric acid (c=3.0 mol/L), the mixed solution was extracted with ether acetone mixture. The total quercetin, kaempferol and isorhamnetin in plasma samples were determined by HPLC, pharmacokinetic parameters were calculated by DAS 3.0 software.

RESULTSThe method was linear over the concentration ranges of 0.0545-8.70, 0.0954-14.7 and 0.0545-8.55 μg/ml for quercetin, kaempferol and isorhamnetin, respectively (r=0.9979, 0.9993, 0.9981). The absolute recoveries were 85.3%-86.1%, 79.4%-86.7% and 62.8%-89.7%, respectively and the assay recoveries were all from 94.7% to 107%. The relative standard deviation (RSD) of intra-and inter-day were less than 9.5% and 9.8%, respectively. The main pharmacokinetic parameters were as follows: T(1/2z) was 92.7, 67.9 and 54.2 h; Tmax was 0.400, 0.400 and 3.87 h; AUC(0-∞) was 68.0, 67.5 and 32.8 mg/h/L; MRT(0-∞) was 128, 85.2 and 72.0 h for quercetin, kaempferol and isorhamnetin, respectively.

CONCLUSIONThe method established in this study is accurate, reliable and reproducible, and can be applied for determination of total quercetin, kaempferol and isorhamnetin in rat plasma after oral administration of FME; the pharmacokinetic studies showed that the distribution of drugs is rapid and elimination is very slow.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Flavonols ; blood ; pharmacokinetics ; Kaempferols ; blood ; pharmacokinetics ; Male ; Plant Extracts ; pharmacokinetics ; Quercetin ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the safety and feasibility of the total laparoscopic anastomosis in laparoscopic gastrectomy.

METHODSClinical data of 36 patients who received totally laparoscopic anastomosis and another 47 patients who received anastomosis through small incision in our department from July 2012 to July 2013 were retrospectively analyzed. Clinical outcomes were compared between the two groups.

RESULTSThe operation was successfully carried out in all the 83 patients. The mean incision length was (7.1±0.9) cm in small incision group and (2.6±0.4) cm in totally laparoscopic group, while the mean time of anastomosis was (70.9±9.0) min and (29.1±4.9) min respectively. Six patients felt moderate pain and 41 felt severe pain in small incision group, while 29 patients felt moderate pain and 7 felt severe pain in totally laparoscopic group. Anastomotic leakage occurred in 1 case after operation in small incision group and there was no related anastomosis complication in totally laparoscopic group.

CONCLUSIONSTotal laparoscopic anastomosis is safe and feasible in laparoscopic gastrectomy for gastric cancer. Compared with small incision-assisted anastomosis, totally laparoscopic anastomosis is associated with shorter time and less pain.

Adult ; Aged ; Anastomosis, Surgical ; methods ; Female ; Gastrectomy ; Gastroenterostomy ; methods ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo study the feasibility and influence of vagus nerve preservation in radical operation for proximal gastric cancer.

METHODSThirty-two patients with early or T2 cardia cancer from May 2007 to May 2009 were enrolled and randomized into two groups, i.e. vagus nerve preservation group(n=16) and control group(n=16). Two groups were compared with regard to operative time, anastomotic fistula, digestive discomforts, body weight, survival rate, findings on gastroscope and abdominal ultrasonography.

RESULTSThere were no statistically significant differences between the two groups in operative time (2.8 vs. 2.5 h), postoperative complications rate (25.0% vs. 31.3%). No recurrence or mortality was observed after one-year follow-up. However, patients who underwent vagus nerve preservation had less postprandial discomforts(3 vs. 12 cases), bile reflux(3 vs. 10 cases), atrophic gastritis(1 vs. 9 cases), gallstones(1 vs. 8 cases), body mass index, and diarrhea(P<0.05).

CONCLUSIONFor patients with early gastric cancer, preservation of the vagus nerve during radical gastrectomy results in less complications and does not compromise patient survival.

Adult ; Aged ; Cardia ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prospective Studies ; Stomach Neoplasms ; surgery ; Vagus Nerve ; surgery

OBJECTIVETo study the expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.

METHODSA tissue microarray was constructed using 85 gastric carcinoma and 25 normal gastric mucosa specimens. The expression of Smo and Gli1 proteins were detected immunohistochemically and the correlation between their expression in gastric carcinoma was analyzed.

RESULTSOnly weak expression, if any, of Smo and Gli1 proteins was detected in normal gastric mucosa, but in papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, their expressions were significant increased as the differentiation degree was lowered. Smo protein expression in gastric carcinoma was significantly correlated with that of Gli1 protein with correlation coefficient of 0.989 (P<0.001).

CONCLUSIONThe abnormal activity of Sonic hedgehog signal transduction pathway may play an important role in the occurrence of papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, and this abnormality is associated with Smo protein overexpression, which upregulates the expression of the downstream transcription factor Gli1 protein.

Adenocarcinoma ; metabolism ; pathology ; physiopathology ; Adult ; Aged ; Female ; Hedgehog Proteins ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; Smoothened Receptor ; Stomach Neoplasms ; metabolism ; pathology ; physiopathology ; Transcription Factors ; biosynthesis ; Zinc Finger Protein GLI1

OBJECTIVETo explore the possibility of repairing chemically induced acute hepatic injuries with allogeneic bone marrow stem cell (BMSC) transplantation.

METHODSA SD rat model of CCl(4)-induced acute hepatic injury was established, which received transplantation of BMSCs (2.0 ml, 1x10(6)/ml) or normal saline injection into the local liver parenchyma, respectively. The rats were sacrificed at 6 h before and 6 h, 1, and 5 weeks after transplantation, and the livers were prepared for microscopic examination.

RESULTSCellular necrosis, bridging necrosis, congestion in the hepatic sinusoid, and inflammatory cell infiltration were seen in the chemically injured livers 6 h after model establishment, and these changes were ameliorated in rats receiving BMSC transplantation.

CONCLUSIONSAllogeneic BMSC transplantation can repair chemically induced acute liver injuries.

Animals ; Bone Marrow Cells ; cytology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; surgery ; Hematopoietic Stem Cell Transplantation ; methods ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Treatment Outcome