Animals ; Antioxidants ; physiology ; Dehydroepiandrosterone ; physiology ; Lipid Metabolism ; physiology ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P

OBJECTIVETo observe the effect of Xinfeng Capsule (XFC) on ankylosing spondylitis (AS) patients' symptoms and signs, serum immunoglobulin levels, peripheral blood lymphocyte autophagy protein, autophagy gene, and to explore its mechanism.

METHODSTotally 59 AS patients were assigned to the treatment group (39 cases) and the control group (20 cases) according to random digit table. Patients in the treatment group received XFC, 0.5 g each pill, three pills each time, 3 times per day, while those in the control group received sulfasalazine (SASP), 0.25 g per tablet, 4 tablets each time, twice per day. Three months consisted of one therapeutic course. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) were statistically calculated. Serum immunoglobulins (IgG1, IgG2, IgG3, IgG4, IgA , SIgA, and IgM) were detected using ELISA. Changes of Beclin1, LC3-II, phosphatidylinositol 3-kinase (PI3K), Akt, the mammalian target of rapamycin (mTOR) were detected using Western blot. Serum autophagy related genes such as Atg1, Atg5, Atg12, Atg13, and Atg17 were detected using the polymerase chain reaction (PCR). The correlation between immunoglobulin subtypes and autophagy gene in AS patients using Spearman correlation.

RESULTSCompared with before treatment, BASDAI, IgG1, lgG3, and IgA decreased (P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.01); ATG1, ATG12, ATG13, and ATG17 mRNA expressions decreased, ATG5 mRNA expression increased (P < 0.01) in the treatment group. But BASDAI, IgG1, and IgA levels decreased (P < 0.05, P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.05); ATG1 and ATG13 mRNA expressions decreased (P < 0.05, P < 0.01) in the control group. Compared with the control group, BASDAI, IgG1, and IgA levels decreased (P < 0.05); PI3K, Akt, mTOR protein expressions decreased (P < 0.01); ATG12 and ATG17 mRNA expression decreased, ATG5 mRNA expression increased (P < 0.01) in the XFC group. Correlation analysis showed AS patients' IgG1, IgG2, IgG3, IgA, SIgA, IgM had negative correlation with ATG17; IgG4 and ATG17 were positively correlated (P < 0.05, P < 0.01).

CONCLUSIONXFC could elevate clinical efficacy of AS patients and enhance their autophagy, which might be achieved by acting on PI3K/Akt/mTOR signal, affecting autophagy gene and autophagy protein expression, taking part in the regulation of proliferation and differentiation of lymphocyte B, and strengthen humoral immunity.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; Beclin-1 ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Lymphocytes ; drug effects ; Membrane Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Spondylitis, Ankylosing ; drug therapy ; Sulfasalazine ; therapeutic use ; TOR Serine-Threonine Kinases ; metabolism

Objective To investigate the effect of 131I on apoptosis of thyrocytes in patients with Graves disease. Methods Forty-seven patients with Graves disease were divided into two groups, two week group (G2w) and four week group (G4w). All patients underwent thyoid needle biopsy before 131I treatment and the repeated biopsy at two weeks (G2w) or four weeks (G4w) after 131I treatment. The positive units of pro-apoptotic proteins (Fas, FasL) and anti-apoptotic protein (Bcl-2) were studied with immunohistochemistry staining. The differences of the two groups were compared with t-test. Liner correlation analysis was applied to study the correlation between 131I dose and apoptosis-related proteins and that between serum sTSH after 131I treatment and apoptosis-related proteins. Results Fas, FasL and Bcl-2 expression (positive units) were significantly increased in both groups after 131I treatment, G2w :22.84 ± 9.31 vs 16.20 ± 6.75,21.13±6.29vs 14.56±4.06, 21.69±7.83 vs 15.22 ±5.94, t= -3.08, -3.73, -4.05 (allP<0.05); G4w:21.69 ±4.52 vs 15.83 ±5.03, 19. 11 ±3.75 vs 14.02 ±4.98, 19.06 ±3.44 vs 16.63 ±4. 73, t = - 5.26, - 5.00, - 2.41 (all P<0.05). However, no statistical differences were found between G2w and G4w (t = 0. 53, 0. 82, 1.46, all P > 0.05). Significant correlation was found between 131I 0. 727, rFasL = 0. 763 (both P<0.05)), but not between the dose and Bcl-2, rBcl-2 = - 0. 094, 0. 102(both P > 0.05). There were significant correlation between serum sTSH three months after 131I treatment and apoptosis-related proteins, rFas = 0.433, rFasL = 0. 601, rBcln2 = - 0. 397, (all P<0. 05). Conclusions 131I can induce thyrocytes to express the pro-apoptotic proteins in patients with Graves disease.

OBJECTIVETo evaluate the association between two single nucleotide polymorphisms located in the promoter of transforming growth factor-β1 receptor 2 (TGFBR2) gene and hypertension in Han Chinese population.

METHODSThe subjects were recruited from the population of cluster sampling survey for essential hypertension (EH) in two townships of Yixing city, Jiangsu province in 2009. Overall, 2012 patients with hypertension and 2116 age (± 2 years) and sex-matched unrelated controls were selected. Epidemiological data, physical measurements results and serum glucose and lipid biomarker were collected and detected. Linkage disequilibrium (LD) analysis were applied and two tagging single nucleotide polymorphisms (tagSNP) in 5' upstream of TGFBR2 gene (rs6785358, -3779A/G; rs764522, -1444C/G) were selected for genotyping and analyzing for the association with hypertension.

RESULTSThe frequencies of AA, AG, GG in case and control of rs6785358 were 1455 (72.3%), 517 (25.7%), 40 (2.0%) and 1582 (74.8%), 490 (23.2%), 43 (2.0%) respectively, and CC, CG, GG of rs764522 were 1524 (75.7%), 464 (23.1%), 24 (1.2%) and 1654 (78.2%), 436 (20.6%), 26 (1.2%) respectively. SNP rs764522 was significantly associated with EH and OR (95%CI) were 1.17 (1.01 - 1.36) (P < 0.05) in dominant model after adjustment for confounding factors such as age, sex, glucose, lipids, smoking and alcohol drinking. Further stratification analysis by age, sex, smoking and alcohol drinking indicated that individuals carrying G allele (CG/GG genotype) of SNP rs764522 had higher susceptibility to EH than CC genotype (OR = 1.21, 95%CI: 1.01 - 1.45) (P < 0.05) in ≥ 55 years group. No statistical significance was detected in the distribution of genotypes and allele frequencies for SNP rs6785358 between cases and controls (P > 0.05). Haplotype analysis showed that no significant frequency difference of haplotype structured by rs6785358 and rs764522 was found between cases and controls (P > 0.05), and no significant blood pressure change was found between genotype variations of rs6785358 and rs764522 (P > 0.05).

CONCLUSIONSNP rs764522 of TGFBR2 gene is associated with increased risk of EH in elderly Han Chinese population.

Aged ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, Transforming Growth Factor beta ; genetics

The female sex is traditionally considered a favorable prognostic factor for nasopharyngeal carcinoma (NPC). However, no particular study has reported this phenomenon. To explore the prognostic impact of gender on patients with NPC after definitive radiotherapy, we reviewed the clinical data of 2063 consecutive patients treated between 1st January 2000 and 31st December 2003 in the Sun Yat-sen University Cancer Center. The median follow-up for the whole series was 81 months. The female and male patients with early stage disease comprised 49.4% and 28.1% of the patient population, respectively. Both the 5-year overall survival (OS) and disease-specific survival (DSS) rates of female patients were significantly higher than those of male patients (OS: 79% vs. 69%, P < 0.001; DSS: 81% vs. 70%, P < 0.001). For patients with locoregionally advanced NPC, the 5-year OS and DSS rates of female vs. male patients were 74% vs. 63% (P < 0.001) and 76% vs. 64%, respectively (P < 0.001). A multivariate analysis showed that gender, age, and TNM stage were independent prognostic factors for the 5-year OS and DSS of NPC patients. The favorable prognosis of female patients is not only attributed to the early diagnosis and treatment but might also be attributed to some intrinsic factors of female patients.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Chemotherapy, Adjuvant ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; drug therapy ; pathology ; radiotherapy ; Neoplasm Staging ; Prognosis ; Radiotherapy, High-Energy ; Sex Factors ; Survival Rate ; Young Adult

Objective Aim of this paper was to explore the trend and characteristics of cancer incidence in 11 areas (5 cities and 6 counties) in China. Methods Data from cancer registries during 1988 to 2002 collected from the 11 cancer registry points were used to analyze the trends and characteristics of cancer incidence rates. Results There were 695 050 newly developed cancer cases in this study. The crude rate of incidence and the world age-adjusted incidence were 215.50/105 and 170.97/105 respectively. The leading cancer sites were lung, stomach, liver, esophagus, breast, colon, rectum, pancreas, bladder and leukemia. The sixteen key cancers accounted for 85.56% of all the cancer cases. The crude incidence rate of all cancers had been significantly increased from 1988 to 2002. Among them, prostate (185.48%) ranked the fastest growing one followed by cancers of the gallbladder, breast, colon, ovarian, lymphoma, bladder, pancreas, rectum, lung, leukemia and liver. The one that had reduced the most was cervix uteri (17.00%), followed by esophagus, stomach and nasopharynx. Conclusion Crude cancer incidence rate increased in the 11 areas in China from 1988 to 2002. The ranking of pancreas cancer, bladder cancer and leukemia came into the top ten. Even though the incidence rates of prostate and gallbladder cancer were relative low but had a fast increase. The results of this study provided a scientific base for the development of a better strategy on cancer prevention and control in China.

OBJECTIVETo investigate the incidence and mortality of colorectal cancer in China from 1998 to 2002, and to analyze its prevalence trend.

METHODSThe cancer registration data in 10 cities and counties in China during the period of 1988-1992, 1993-1997 and 1998-2002 were used to investigate the incidence, mortality, and prevalence trend of colorectal cancer from 1988 to 2002.

RESULTSThe total number of new cases of colorectal cancer in the 10 cities and counties during 1988-2002 was 62,793, accounting for 9.27% of all malignant tumors. The crude incidence rate was 20.10/10(5), and the age-standardized incidence adjusted by world population was 15.63/10(5). The total number of death of colorectal cancer in the 10 cities and counties during 1988-2002 was 35,545, accounting for 7.37% of all malignant tumors. The mortality rate was 11.38/10(5), the age-standardized mortality rate adjusted by world population was 8.70/10(5). The incidence and mortality of colorectal cancer during 1988-2002 increased by 38.56% and 15.30%, respectively, and the incidence and mortality rates in urban area was higher than that in rural area, and higher in males than in females. The crude incidence rate of colon cancer was higher than that of rectal cancer, especially in urban area, but the mortality of rectal cancer was a little bit higher than that of colon cancer.

CONCLUSIONThere is an increasing trend in both the incidence and mortality rates of colorectal cancer from 1988-2002 in the 10 cities and counties in China. Measures should further be taken in the prevention and treatment of colorectal cancer in the whole population of China in future.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Cities ; Colonic Neoplasms ; epidemiology ; mortality ; Colorectal Neoplasms ; epidemiology ; mortality ; Female ; Humans ; Incidence ; Infant ; Male ; Middle Aged ; Rectal Neoplasms ; epidemiology ; mortality ; Rural Population ; Sex Factors ; Urban Population ; Young Adult

Soy isoflavones have been reported to be natural chemopreventive in several types of human cancer. Daidzein and genistein are two main components of soy isoflavones. In our previous study, they were shown to be anti-proliferative and induce cell cycle arrest at S phase of SHZ-88 rat breast cancer cells. We hypothesized that soy isoflavones might exert its anticancer effect by activating cAMP/PKA pathway. The present study was designed to analyze the effect of soy isoflavones on the cAMP/PKA pathway in SHZ-88 cells. Daidzein and genistein were dissolved in DMSO. Cells were treated with 50 mug/ml daidzein and 15 mug/ml genistein, respectively, and with only equal DMSO in the culture medium as control. The cellular cAMP content was tested by radioimmunoassay (RIA). The activity of adenylate cyclase (AC), phosphodiesterase (PDE) and PKA were measured by RIA and (gamma-(32)P) ATP incorporation. Reverse transcript-polymerase chain reaction (RT-PCR) was used to analyze the expression of cAMP response element binding protein (CREB) mRNA of the cells. The results showed that the concentration of cAMP in the cells treated with 50 mug/ml daidzein and 15 mug/ml genistein was significantly increased by 9.5%and 11.0%, respectively, 5 min later (P<0.05), then increased by 31.0%and 40.3%, respectively, 10 min later (P<0.01), compared with that of the control group cells. The activity of AC was not affected during the course of experiment, but that of PDE was decreased to 71.8%and 71.6%, respectively, in the control group 5 min later (P<0.05). The PKA activity was increased to 125.8%and 122.3%, respectively, in the control group 20 min after the cells were treated with daidzein and genistein (P<0.05), and kept at high level till 40 min after treatment. CREB mRNA of the cells treated with daidzein and genistein was increased by 31.6%and 51.1%, respectively, 3 h later (P<0.05), then began to decrease 6 h after treatment. The current study suggests that soy isoflavones activate the cAMP/PKA pathway in SHZ-88 rat breast cancer cells by inhibiting the activity of phosphodiesterase.

The present study aimed to investigate the effect of proto-oncogene c-src on the viability of rat spermatogonial stem cells from 9 day-old rat in vitro. MTT method was used to observe the viability of the spermatogonial stem cells treated with antisense c-src oligodeoxynucleotides (ODNs) in vitro; RT-PCR was utilized to observe the expression of c-src mRNA and Western blot was used to observe the protein expressions of pp60c-src and phosphorylated signal transducer and activator of transcription-3 (p-STAT3). Compared with that in control group, the viability of spermatogonial stem cells decreased by 8.1% (P<0.05) and the expression of c-src mRNA decreased significantly after treatment with 10 μmol/L antisense c-src ODNs for 12 h. Compared with that in the control group, the protein expressions of pp60c-src and p-STAT3 decreased by 33.8% and 45.3% (both P<0.01), respectively, in the spermatogonial stem cells after being transfected with antisense c-src ODNs. The results suggest that proto-oncogene c-src regulates the viability of rat spermatogonial stem cells through p-STAT3.
Animals ; Cells, Cultured ; Genes, src ; Male ; Phosphorylation ; Proto-Oncogene Proteins pp60(c-src) ; metabolism ; RNA, Messenger ; Rats ; STAT3 Transcription Factor ; metabolism ; Spermatogonia ; cytology ; metabolism ; Stem Cells ; cytology ; metabolism ; Transfection