Background:As an important catalytic subunit of telomerase,human telomerase reverse transcriptase (hTERT)plays an important role in the development and progression of many cancers including gastric cancer.It has been reported that several single nucleotide polymorphisms (SNPs)of hTERT had varying degrees of association with risk of neoplasms. Aims:To study the correlation between SNPs of hTERT rs2853676 and rs2853677 and susceptibility to gastric cancer. Methods:Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotypes of rs2853676 and rs2853677 of hTERT in 297 gastric cancer patients,105 atrophic gastritis and 402 controls. Helicobacter pylori (Hp)infection was detected by pathological examination and 13 C-urea breath test.Results:Frequency of AA genotype of rs2853676 was significantly higher in gastric cancer group when compared with control group (15.2%vs.6.5%,P =0.01).The risk of gastric cancer in AA genotype carriers increased 2.47-fold (95% CI:1.46-4.16) when compared with GG carriers.No significant differences in the frequencies of CC,TC and TT genotypes of rs2853677 were found among gastric cancer patients,atrophic gastritis patients and controls.Hp infection rates in atrophic gastritis group and gastric cancer group were significantly increased than those in controls (64.8%,56.9% vs.40.3%,P all <0.01),OR were 2.73 (95% CI:1.74-4.26),1.96 (95% CI:1.44-2.67),respectively.Logistic regression analysis showed that there was no significant interaction between Hp infection and gene mutation.Conclusions:Polymorphism of hTERT gene rs2853676 may play a role in susceptibility to gastric cancer,and Hp infection may not be involved in the increase of risk of gastric cancer caused by hTERT gene polymorphism.

Objective To analyze the drug resistant characteristics of 84 clinical isolated Helicobacter pylori (Hp) strains, and to observe the inhibitory effects of anti-Hp Lactobacillus acidophilus (La)4 and La6 on different antibiotic-resistant Hp strains. Methods Hp strains were isolated and cultured from gastric mucosa of 84 different gastropathy patients (20 patients with chronic gastritis, 24 with gastric ulcer, 19 with duodenal ulcer and 16 with gastric cancer). The minimum inhibitory concentration (MIC) of metronidazole, clarithromycin and amoxicillin were tested by E-test in order to determine the resistance of these three antibiotics in clinical isolated Hp strains. With standard La as control, the supernatant of anti-Hp La4 and La6 was added into Hp strains culture wells. Hp strains were cultured in solid media for 72 hours, and then inhibition ring were recorded. Anti-Hp Lactobacillus acidophilus liquid was also added to culture medium of different Hp strains, which were in liquid culture, culture medium were taken at different time points (4,8,12,24,48 hrs) to calculate bacteria colony number and test urease activity. Results In 84 clinical isolated Hp strains, the resistant rates of metronidazole, clarithromycin and amoxicillin resistance rates were 67.9%, 17.9% and 1.2% respectively. Of those 11 strains were mixed drug resistance, which included 10 strains of metronidazole and clarithromycin mixed drug resistance, and one of metronidazole and amoxicillin mixed drug resistance. In solid culture conditions, supernatant of anti-Hp Lactobacillus acidophilus La4 and La6 had obvious inhibitory effect on antibiotic-resistant and non-resistant Hp strains. In liquid culture conditions, anti-Hp Lactobacillu acidophilus La4 and La6 bacterium liquid could inhibit the proliferation of antibiotic-resistant and non-resistant Hp strains, the antagonistic role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). The urease activity of antibiotic-resistant Hp strains was inhibited since mixed cultured with anti-Hp Lactobacillu acidophilus La4 and La6 for 4 hours, the urease activity gradually decreased as culture time extended, and the inhibitory role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). Conclusions In 84 Hp strains, most were metronidazole resistant strains, followed by clarithromycin resistant strains, metronidazole and clarithromycin mixed resistance strains. In vitro, anti-Hp Lactobacillu acidophilus La4 and La6 had obvious inhibitory effects on antibiotic-resistant and non-resistant Hp strains.

Objective To investigate the effect of resveratrol on the expression of peroxisome proliferator activated re-ceptor γ coactivator-1α(PGC-1α)in skeletal muscle of rats with chronic obstructive pulmonary disease(COPD). Methods A total of 45 male Sprague Dawley rats were randomly divided into control group,model group and resveratrol group,15 rats in each group. The rats in the model group and resveratrol group were made COPD model through the intratracheal instillation of lipopolysaccharide and repeated smoke exposure,except the rats in the control group. From the 29th day of smoke exposure,the rats in the control group and model group were given 2 mL saline by gavage,once a day for 30 days;and the rats in the resvera-trol group were given 2 mL resveratrol solution by gavage(100 mg·kg - 1 ·d - 1 ),once a day for 30 days. After 30 days of con-tinuous gavage,the rats were sacrificed,then arterial blood and skeletal muscles were harvested. The level of tumor necrosis factor-α(TNF-α)in serum and skeletal muscle of rats was detected by enzyme linked immunosorbent assay. The expression of PGC-1α,nuclear respiratory factor 1(NRF1),mitochondrial transcription factor A(Tfam)and cytochrome C oxidase Ⅳ(COXⅣ)mRNA in skeletal muscle tissues of rats was determined by quantitative real-time polymerase chain reaction. The expres-sion of PGC-1α,NRF1,Tfam and COX Ⅳ protein was detected by Western blot. Results The level of TNF-α in serum and skeletal muscle tissues of rats in the model group and resveratrol group was significantly higher than that in the control group (P < 0. 01). The level of TNF-α in serum and skeletal muscle tissues of rats in the resveratrol group was significantly lower than that in the model group(P < 0. 01). The expression of PGC-1α,NRF1,Tfam,COX Ⅳ protein and mRNA in skeletal mus-cle tissues of rats in the resveratrol group and model group was significantly lower than that in the control group(P < 0. 01). The expression of PGC-1α,NRF1,Tfam,COX Ⅳ protein and mRNA in skeletal muscle tissues of rats in the resveratrol group was significantly higher than that in the model group(P < 0. 01,P < 0. 05). Conclusion Resveratrol can reduce the level of TNF-α in serum and skeletal muscle tissues of COPD rats,increase the expression of PGC-1α,thereby improving the mitochon-drial biosynthesis function.

OBJECTIVETo investigate the clinicopathologic features of calcium pyrophosphate dihydrate crystal deposition disease (CPPD-CDD).

METHODSThe clinical and pathologic profiles were retrospectively analysed in 20 cases of CPPD-CDD.

RESULTSCPPD-CDD was far more common in women, most frequently involving joints, especially the knees and presenting with various arthrisis. Abnormally calcified and the articular damages were characteristic features by imageing. Histologically, multifocal indigo granular calcinosis was seen in synovium and sometimes appeared as needle-shaped or rhomboid crystals, which characterized the CPPD.

CONCLUSIONSThough clinical symptoms of CPPD are quite variable, the definite diagnosis can be made by the abnormal calcification and joint damage radiographically and the indigo CPPD crystals histopathologically.

Adult ; Aged ; Aged, 80 and over ; Chondrocalcinosis ; diagnostic imaging ; pathology ; surgery ; Female ; Follow-Up Studies ; Hip Joint ; diagnostic imaging ; pathology ; surgery ; Humans ; Intervertebral Disc ; diagnostic imaging ; pathology ; surgery ; Knee Joint ; diagnostic imaging ; pathology ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Spinal Diseases ; diagnostic imaging ; pathology ; surgery ; Synovial Membrane ; pathology ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of the photodynamic therapy (PDT) with the new water-soluble metalloporphyrin compound on human prostate cancer PC-3 cells in vitro and the anticancer mechanism of PDT.

METHODSThe new water-soluble manganese, 5,10,15, 20-tetra (N-methyl4-pyridyl) porphinato (2-) tetraiodide salt, was synthesized. The PC-3 cells were treated with the compound of serial concentrations(0, 0.1, 1, 1.0 micromol/L) followed by irradiation of different dosages of visible light. The techniques of MTT and Annexin-V/propidium iodide double-labeled flow cytometry (FCM) were applied to measuring the inhibitory effect of the compound on the growth activity and apoptosis of the cells.

RESULTSWhen the metalloporphyrin compound concentration was within 10 micromol/L and the irradiation time was within 30 min, the water-soluble metalloporphyrin compound had a significant inhibitory effect on the proliferation of PC-3 cells and induced PC-3 cell apoptosis, and the effects depended greatly on metalloporphyrin concentration and illumination dosages. Higher concentrations and dosages induced the death of the majority of PC-3 cells.

CONCLUSIONThe PDT of the water-soluble metalloporphyrin compound followed by light irradiation has a distinctive killing effect on PC-3 cells in vitro, and the rates of proliferation inhibition and cell apoptosis are correlated with metalloporphyrin concentration and the dosages of light irradiation. The results suggest that the mechanism of metalloporphyrin PDT may be involved with the induction of apoptosis in human prostate cancer cells.

Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Dose-Response Relationship, Radiation ; Humans ; Male ; Metalloporphyrins ; pharmacology ; Photochemotherapy ; Prostatic Neoplasms ; pathology

OBJECTIVETo investigate the possible mechanisms of miR-21-mediated regulation of proliferation and activation of hepatic oval cells.

METHODSThe 2-acetamidofluorene/partial hepatectomy (2-AAF/PH) method was applied to generate hepatic oval cell activation model in male Sprague-Dawley rats; after the 7 days of 2-AAF/PH or PH alone (control), the rats were sacrificed at 0 h, 6 h, 12 h, 24 h, 72 h and 168 h. Expression of miR-21 was detected by real-time PCR and differences between groups were evaluated using the two-sample t-test. Differential transcription of miR-21 target genes was assessed bioinformatically, and with western blotting to detect changes in protein expression of the target gene.

RESULTSThe rat hepatic oval cell activation model was successfully established.The 2-AAF/PH rats showed miR-21 expression beginning to increase at 12 h, peaking at 24 h, and decreasing thereafter until an increase at 168 h.For the control group, the miR-21 expression began to increase at 6 h, until 24 h when expression began steadily declining to reach the original level.Compared to the control group, the experimental group showed expression of miR-21 that was significantly less at 6 h (P=0.039, t =3.029) and significantly more at 24 h and 168 h (P=0.026, t =-3.433 and P=0.007, t =-5.105). Among the predicted target genes of miR-21 were WW domain containing E3 ubiquitin protein ligase 1 (WWD), Smad family member 7 (Smad7), and polybromo-1 (Pbrm1).Smad7 protein expression began to decrease at 6 h in the control group, until reaching its minimum at 24 h when it increased; in the experimental group, SMAD7 expression increased at 6 h, then began to decrease with the minimum detected at 168 hour.In the control group, the Smad7 mRNA expression decreased slightly at 6 h, then began to increase, reaching its peak at 24 h when the expression fell to the original level. In the experimental group, the Smad7 mRNA expression began to increase at 6 h and reached its peak at 24 h when it decreased; the expression was little more than its original level at 168 h.Smad7 protein expression was negatively correlated with miR-21, and Smad7 mRNA expression was positively correlated with miR-21 but negatively correlated with Smad7 protein expression.

CONCLUSIONmiR-21 may play a vital role in the activation and proliferation of hepatic oval cells.As a target gene of miR-21, Smad7 might be involved in the process.

2-Acetylaminofluorene ; Animals ; Cell Proliferation ; Hepatectomy ; Hepatocytes ; cytology ; Liver ; cytology ; Male ; MicroRNAs ; genetics ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

OBJECTIVETo study the effects of different immunodepressants on the sperm parameters of kidney transplant recipients.

METHODSIn 15 healthy fertile men and 37 kidney transplant recipients, ejaculates were aseptically obtained by masturbation. Thirty-seven patients were divided into two groups, 20 patients were treated with Prograf (FK506) combination with mycophenolate mofetil (MMF) and prednisone; 17 patients were treated with cyclosporine (CsA) combination with azathioprine with prednisone. The sperm viability, mobility parameters such as prorsad percentage motility, straight line velocity (VSL), curve line velocity (VCL), velocity of average path (VAP) and morph were estimated with a computer-assisted sperm analyzer (CASA) provided with a multiple-exposure photography system.

RESULTSThere were no significant difference in sperm viability rate [(81.7 +/- 5.7)%, (79.4 +/- 6.8)% and (83.8 +/- 6.0)%], VCL [(24.1 +/- 8.6)%, (23.9 +/- 4.4)%, (24.8 +/- 4.2)% ] and VAP [(19.7 +/- 6.6)%, (18.6 +/- 2.9)%, (21.0 +/- 4.0)%] among groups of FK506, CsA and control, respectively (P > 0.05). The rate of anomaly [(67.8 +/- 5.7)%], the prorsad percentage motility [(46.4 +/- 8.1)%] and VSL [(15.4 +/- 4.6)%] in the group of FK506 were respectively significantly lower and higher than those in the group of CsA [(80.1 +/- 5.6%, (33.3 +/- 6.4)%, (10.2 +/- 2.4)%] (P < 0.05).

CONCLUSIONThe application of FK506 combined with MMF could help recover the mobility and morphology of the sperm in kidney transplantation recipients.

Adolescent ; Adult ; Case-Control Studies ; Cyclosporine ; pharmacology ; Drug Therapy, Combination ; Humans ; Immunosuppressive Agents ; pharmacology ; Kidney Transplantation ; Male ; Mycophenolic Acid ; analogs & derivatives ; pharmacology ; Prednisone ; pharmacology ; Sperm Motility ; drug effects ; Tacrolimus ; pharmacology

OBJECTIVETo investigate the effects of the small interfering RNA plasmid-dextran magnetic nanoparticles (siRNA-DMN) combination with external magnetic fields on silencing survivin gene expression of bladder cancer cells and apoptosis when DMN used as gene carrier to transfer siRNA-survivin recombinant plasmid in vivo.

METHODSThe siRNA-survivin recombinant plasmid specific targeted survivin was synthesized in previous experiment. DMN were prepared by chemical coprecipitation method and used as gene carrier. The siRNA-DMN were constructed by static electricity of polylysine and transferred into human bladder cancer BIU-87 cells with the help of external magnetic fields. The growth inhibiting rate (IR) of BIU-87 cells was observed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide's test and the apoptosis index (AI) was detected by transferase-mediated dUTP nick end labeling method. The relatively transcription levels of survivin mRNA and protein expression were respectively detected by semi-quantitive reverse transcription polymerase chain reaction and Western Blotting techniques.

RESULTSThe diameter, effective diameter and saturation magnetization of DMN-siRNA were about 10 - 12 nm, 94.8 nm and 0.19 emu/g, respectively. The IR (39.60%) and AI (28.72%), the relative expression of survivin mRNA and protein of siRNA-DMN combination with external magnetic fields on BIU-87 cells were significantly higher and lower than those in the control group and single siRNA-DMN group, respectively (P < 0.05).

CONCLUSIONSThe siRNA-survivin plasmid-DMN combination with external magnetic fields could effectively inhibit survivin expression and induce BIU-87 cells apoptosis which provided experimental basis for the magnetic targeting gene therapy of bladder tumor.

Apoptosis ; drug effects ; Cell Line, Tumor ; Electromagnetic Fields ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Glucans ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Nanoparticles ; Neoplasm Proteins ; genetics ; Plasmids ; RNA, Messenger ; genetics ; RNA, Small Interfering ; pharmacology ; Transfection ; methods ; Urinary Bladder Neoplasms ; genetics ; pathology ; therapy

Objective To determine the efficacy of temozolomide (TMZ) in treating patients with glioblastoma using carmustine (BCNU) as control, and examine the effect of O6-methylguanine-DNA methyltransferase (MGMT) expression on the prognosis of patients with glioblastoma. Methods Two hundred and eighty-three patients with pathologically confirmed glioblastoma,admitted to and received treatment without chemotherapeutics in our hospital from January 2004 to January 2009, were enrolled in our study; TMZ was used in 97 patients and BCNU in 186 patients.The glioma tissues were examined for MGMT protein expression by immunohistochemistry.All patients in these 2 groups were performed long-term follow-up for survival time of the patients, tumor response and drug safety. Results In patients of the TMZ treatment group,the median survival time was (19.2±0.6) months,the 2-year survival rate 31％ (30/97),the 5-year survival rate 6.2％ (6/97) and the objective response rate 75.26％ (73/97); while in patients of the BCNU treatment group,the median survival time of was (15.6±0.6) months,the 2-year survival rate 14％ (26/186),the 5-year survival rate 0.5％ (1/186),and objective response rate 45.16％ (84/186); the cumulative survival rate of patients received TMZ treatment was significantly higher than that of patients received BCNU treatment (P＜0.05);the objective response rate between the 2 groups was obviously different (x2=24.753, P=0.000); the incidence of hypoleukocytosis in patients received TMZ treatment was significantly lower than that in patients received BCNU treatment (x2=15.681,P=0.000). Conclusion TMZ shows more efficient oobjective response as compared with BCNU, with less adverse reaction and better tolerability.Therefore,it is an ideal drug of postoperative adjuvant chemotherapy for malignant glioma.

Background: According to the 7th edition of the American Joint Committee on Cancer (AJCC) staging system, over 50% of patients with nasopharyngeal carcinoma (NPC) have N1 disease at initial diagnosis. However, patients with N1 NPC are relatively under-researched, and the metastasis risk of this group is not well-stratified. This study aimed to evaluate the prognostic values of gross tumor volume of metastatic regional lymph node (GTVnd) and pretreatment serum copy number of Epstein–Barr virus (EBV) DNA in predicting distant metastasis of patients with N1 NPC, and to develop an integrated prognostic model that incorporates GTVnd and EBV DNA copy number for this group of patients. Methods: The medical records of 787 newly diagnosed patients with nonmetastatic, histologically proven N1 NPC who were treated at Sun Yat-sen University Cancer Center between November 2009 and February 2012 were ana-lyzed. Computed tomography-derived GTVnd was measured using the summation-of-area technique. Blood sam-ples were collected before treatment to quantify plasma EBV DNA. The receiver operating characteristic (ROC) curve analysis was used to evaluate the cut-off point for GTVnd, and the area under the ROC curve was used to assess the predicted validity of GTVnd. The survival rates were assessed by Kaplan–Meier analysis, and the survival curves were compared using a log-rank test. Multivariate analysis was conducted using the Cox proportional hazard regression model. Results: The 5-year distant metastasis-free survival (DMFS) rates for patients with GTVnd > 18.9 vs. ≤ 18.9 mL were 82.2% vs. 93.2% (P < 0.001), and for patients with EBV DNA copy number > 4000 vs. ≤ 4000 copies/mL were 83.5% vs. 93.9% (P < 0.001). After adjusting for GTVnd, EBV DNA copy number, and T category in the Cox regression model, both GTVnd > 18.9 mL and EBV DNA copy number > 4000 copies/mL were significantly associated with poor prognosis(both P < 0.05). According to combination of GTVnd and EBV DNA copy number, all patients were divided into low-, moderate-, and high-risk groups, with the 5-year DMFS rates of 96.1, 87.4, and 73.8%, respectively (P < 0.001). Multi-variate analysis confirmed the prognostic value of this model for distant metastatic risk stratification (hazard ratio [HR], 4.17; 95% confidence interval [CI] 2.34–7.59; P < 0.001). Conclusions: GTVnd and serum EBV DNA copy number are independent prognostic factors for predicting distant metastasis in NPC patients with N1 disease. The prognostic model incorporating GTVnd and EBV DNA copy number may improve metastatic risk stratification for this group of patients.