Objective To explore the role of methylation of Human MutS homolog 2 (MSH2) and Human runt-related transcription factor 3 genes (RUNX3) in DNA prepared from nasopharyngeal swabs in early diagnosis and prognosis prediction of nasopharyngeal carcinoma. Methods The methylation-specific PCR was used to detect hypermethylation of MSH2 and RUNX3 genes in DNA prepared from nasopharyngeal swabs from 54 nasopharyngeal carcinoma patients, 18 chronic nasopharyngitis patients and 20 healthy volunteers. The specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma by detecting MSH2 and/or RUNX3 gene methylation were evaluated. The relationship between methylation of MSH2 or RUNX3 gene and the biological behavior of nasopharyngeal carcinoma was analyzed. Results Hypermethylation of MSH2 and RUNX3 gene was respectively detected in 38 out of 54 (70.37%) and in 28 out of 54 (51.85%) nasopharyngeal swabs obtained from nasopharyngeal carcinoma patients, while there was no methylation in nasopharyngeal swabs from 18 chronic nasopharyngitis patients and 20 healthy volunteers. The differences were statistically significant (P<0.001). The specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma were respectively 100% and 70.37% by detecting MSH2 gene methylation, 100% and 51.85% respectively by detecting RUNX3 gene methylation in nasopharyngeal swabs from nasopharyngeal carcinoma patients determined by methylation-specific PCR, while parallel combined detection of MSH2 and RUNX3 gene methylation increased the diagnostic specificity and sensitivity up to 100% and 90.74%, respectively. No close correlation was found between methylation of MSH2 or RUNX3 gene and the biological behavior of nasopharyngeal carcinoma in patients (P>0.05). Conclusions Parallel combined testing of MSH2 and RUNX3 gene methylation in DNA prepared from nasopharyngeal swabs determined by methylation-specific PCR could increase the specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma, and is of important clinical significance. However it may not serve as an index in evaluating the clinical prognosis of nasopharyngeal carcinoma at present.

Objective To explore the role of methylation of Human MutS homolog 2 (MSH2) and Human runt-related transcription factor 3 genes (RUNX3) in DNA prepared from nasopharyngeal swabs in early diagnosis and prognosis prediction of nasopharyngeal carcinoma. Methods The methylation-specific PCR was used to detect hypermethylation of MSH2 and RUNX3 genes in DNA prepared from nasopharyngeal swabs from 54 nasopharyngeal carcinoma patients, 18 chronic nasopharyngitis patients and 20 healthy volunteers. The specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma by detecting MSH2 and/or RUNX3 gene methylation were evaluated. The relationship between methylation of MSH2 or RUNX3 gene and the biological behavior of nasopharyngeal carcinoma was analyzed. Results Hypermethylation of MSH2 and RUNX3 gene was respectively detected in 38 out of 54 (70.37%) and in 28 out of 54 (51.85%) nasopharyngeal swabs obtained from nasopharyngeal carcinoma patients, while there was no methylation in nasopharyngeal swabs from 18 chronic nasopharyngitis patients and 20 healthy volunteers. The differences were statistically significant (P<0.001). The specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma were respectively 100% and 70.37% by detecting MSH2 gene methylation, 100% and 51.85% respectively by detecting RUNX3 gene methylation in nasopharyngeal swabs from nasopharyngeal carcinoma patients determined by methylation-specific PCR, while parallel combined detection of MSH2 and RUNX3 gene methylation increased the diagnostic specificity and sensitivity up to 100% and 90.74%, respectively. No close correlation was found between methylation of MSH2 or RUNX3 gene and the biological behavior of nasopharyngeal carcinoma in patients (P>0.05). Conclusions Parallel combined testing of MSH2 and RUNX3 gene methylation in DNA prepared from nasopharyngeal swabs determined by methylation-specific PCR could increase the specificity and sensitivity in early diagnosis of nasopharyngeal carcinoma, and is of important clinical significance. However it may not serve as an index in evaluating the clinical prognosis of nasopharyngeal carcinoma at present.

Objective To discuss the growth suppressing, apoptosing effect of new type tumor-supressor gene-p33ING1 in stomach cancer cell strain, and to explore new strategies and methods in tumor therapy. Methods The PCDNA3/p33ING1 nuclear expressing microsome was constructed, p33ING1 and wt-p53 were implanted to human stomach cancer cell both and to evaluate the effect of p33ING1 and p53 toward stomach cancer cell and synergism between them. Results The PCDNA3/p33ING1 nuclear expressing microsome was successfully constructed. The human stomach cancer cell strain SSCG-7901 under implantation of p33ING1 and wt-p53 showed a significant decrease in cell growth, the coupling time was delayed, DNA synthetic phase was shortened and G0/G1 phase prolonged. The cell collapse increased. Conclusions Despite of the tumor-inhibiting effect and biochemical activation of p33ING1, it also plays a role with p53 gene in controling growth of stomach cancer cell, inducing cell collapse and hampering cell proliferation cycle. P33ING1 and p53 are synergistic to each other.

Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; Child ; Child, Preschool ; DNA, Viral ; blood ; Female ; Follow-Up Studies ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Recombinant Proteins ; Time Factors ; Treatment Outcome

OBJECTIVETo assess the association of the A260G and A386G single nucleotide polymorphisms (SNP) of the DAZL gene with male infertility in the Chinese population of Zhejiang Province.

METHODSWe collected the peripheral blood samples from 317 idiopathic infertile males with azoospermia or oligozoospermia and 246 normal fertile men, and genotyped the polymorphic loci of the A260G and A386G polymorphisms of the DAZL gene using the SNaPshot technique.

RESULTSThe DAZL gene A260G was found genetically polymorphic in the Chinese population of Zhejiang Province, with the gene frequencies and their distribution consistent to the Hardy-Weinberg equilibrium. The frequencies of the AA, AG and GG genotypes of the A260G polymorphism were 92.3%, 7.3%, and 0.4% respectively in the normal controls and 94.3%, 5.7%, and 0% in the infertile patients, with no statistically significant differences between the two groups (P = 0.43, OR = 0.78, 95% CI 0.413-1.46). Heterozygosis (AG) of A386G was found in 1 of the control males but not in the infertile patients, while homozygosis (GG) of A386G was not observed in either group (P = 0.259, OR = 0.698, 59% CI: 0.374-1.306).

CONCLUSIONA260G and A386G SNPs of the DAZL gene are not associated with spermatogenic failure and neither represents a molecular marker for the genetic diagnosis of male infertility in the Chinese population of Zhejiang Province.

Asian Continental Ancestry Group ; Azoospermia ; genetics ; China ; Gene Frequency ; Genetic Markers ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Oligospermia ; genetics ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; RNA-Binding Proteins ; genetics

Objective To explore the possible mechanisms of Galectin - 1(Gal - 1)protein in promoting the invasion and migration of gastric cancer cells. Methods After treated with different concentrations(0,1,5 μg/ mL)of Gal - 1 protein, the Trans - well model was used to analyze the invasion and migration ability of gastric cancer. WB and gelatin zymography method were used to detect the MMP - 9 expression and active form change in gastric cancer cells after Gal - 1 stimulate, in order to explore the possible molecular mechanisms of Gal - 1 protein in promoting the invasion and migration of gastric cancer cells. Results In cell migration assay,the number of gastric cancer cells BGC - 823 treated with 1and 5 μg/ mL Gal - 1 stimulate were 117 ± 8. 19 and 167 ± 7. 55,higher than that treated with 0 μg/ mL(P < 0. 05). The number of gastric cancer cells 7 901 treated with 1and 5 μg/ mL Gal - 1 stimulate were 151 ± 5. 13 and 190. 3 ± 6. 8,higher than that treated with 0 μg/ mL(P < 0. 05). In cell invasion assay,the number of gastric cancer cells BGC - 823 treated with 1and 5μg/ mL Gal - 1 stimulate were 51 ± 3. 6 and 76. 7 ± 9. 07,higher than that treated with 0 μg/ mL(P < 0. 05). The number of gastric cancer cells 7 901 treated with 1and 5 μg/ mL Gal - 1 stimulate were 74. 0 ± 7. 21 and 105. 3 ± 11. 37,higher than that treated with 0 μg/ mL(P < 0. 05). The migration and invasion level were significantly increased in gastric cancer cells after Gal - 1 stimulate. The MMP - 9 expression level and active form change in gastric cancer cells were also increased after Gal - 1 stimulate. Conclusion Gal - 1cound significantly promote gastric cancer cell migration and invasion by up - regulated the MMP - 9 expression and active its enzyme activity.

OBJECTIVETo observe the effect of Cordyceps sinensis (CS) powder on renal oxidative stress and mitochondria functions in 5/6 nephrectomized rats, and to primarily explore its possible mechanisms.

METHODSTotally 30 male Sprague-Dawley rats were divided into the sham-operation group, the model group, and the treatment group by random digit table, 10 in each group. A chronic kidney disease (CKD) rat model was prepared by one step 5/6 nephrectomy. Rats in the treatment group were intragastrically administered with CS powder solution at the daily dose of 2 g/kg, once per day. Equal volume of double distilled water was intragastrically administered to rats in the sham-operation group and the model group. All medication lasted for 12 weeks. The general condition of rats, their body weight, blood pressure, 24 h proteinuria, urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (SCr) , and blood urea nitrogen (BUN) were assessed before surgery, at week 2, 4, 6, 8, 10, and 10 after surgery. Pathological changes of renal tissues were observed under light microscope. Morphological changes of mitochondria in renal tubular epithelial cells were observed under transmission electron microscope. Activities of antioxidant enzymes including reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), and malondialdehyde (MDA) in fresh renal tissue homogenate were detected. Mitochondria of renal tissues were extracted to detect levels of mitochondrial membrane potential and changes of reactive oxygen species (ROS). And expressions of cytochrome-C (Cyto-C) and prohibitin in both mitochondria and cytoplasm of the renal cortex were also measured by Western blot.

RESULTS(1) Compared with the sham-operation group, body weight was significantly decreased at week 2 (P <0. 01), but blood pressure increased at week 4 (P <0. 05) in the model group. Compared with the model group, body weight was significantly increased at week 12 (P <0. 01), but blood pressure decreased at week 8 (P < 0. 01) in the treatment group. (2) Compared with the sham-operation group, 24 h proteinuria, urinary NAG, blood SCr and BUN significantly increased in the model group (all P <0. 01). Compared with the model group, blood and urinary biochemical indices all significantly decreased in the treatment group (all P <0. 01). (3) Results of pathological renal scoring: Glomerular sclerosis index, scoring for tubulointerstitial fibrosis, degree of tubulointerstitial inflammatory infiltration were all obviously higher in the model group than in the sham-operation group (all P <0. 01). All the aforesaid indices were more obviously improved in the treatment group than in the model group (all P <0. 01). (4) Compared with the sham-operation group, activities of MnSOD and GSH-Px were significantly reduced, but MDA contents obviously increased in the renal cortex of the model group (all P <0. 01). Compared with the model group, activities of MnSOD and GSH-Px obviously increased (P <0. 05, P <0. 01), but MDA contents obviously decreased in the renal cortex of the treatment group (P <0. 01). (5) Compared with the sham-operation group, the mitochondrial membrane potential significantly decreased, but ROS levels significantly increased in the model group (all P <0.01). Compared with the model group, mitochondrial transmembrane potential increased in the treatment group, thereby inhibiting the tendency of increased production of ROS (both P < 0. 01). (6) Results of Western blot showed that, compared with the sham-operation group, expression levels of mitochondrial Cyto-C and Prohibitin were significantly reduced in the renal cortex (P <0. 01), but significantly elevated in the cytoplasm of the model group (P <0. 01). Compared with the model group, each index was obviously improved in the treatment group with statistical difference (P <0. 05, P <0. 01).

CONCLUSIONCS powder had renal protection, and its mechanism might partially depend on in- hibition of oxidative stress and protection for mitochondria.

Acetylglucosaminidase ; metabolism ; Animals ; Blood Urea Nitrogen ; Cordyceps ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; Kidney Cortex ; Kidney Diseases ; Kidney Function Tests ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; Nephrectomy ; Oxidative Stress ; drug effects ; Proteinuria ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Sequential methodology based on the application of three types of experimental designs was used to optimize the fermentation conditions for elastase production from mutant strain ZJUEL31410 of Bacillus licheniformis in shaking flask cultures. The optimal cultivation conditions stimulating the maximal elastase production consist of 220 r/min shaking speed, 25 h fermentation time, 5% (v/v) inoculums volume, 25 ml medium volume in 250 ml Erlenmeyer flask and 18 h seed age. Under the optimized conditions, the predicted maximal elastase activity was 495 U/ml. The application of response surface methodology resulted in a significant enhancement in elastase production. The effects of other factors such as elastin and the growth factor (corn steep flour) on elastase production and cell growth were also investigated in the current study. The elastin had no significant effect on enzyme-improved production. It is still not clear whether the elastin plays a role as a nitrogen source or not. Corn steep flour was verified to be the best and required factor for elastase production and cell growth by Bacillus licheniformis ZJUEL31410.
Bacillus ; enzymology ; growth & development ; Fermentation ; Pancreatic Elastase ; biosynthesis ; Research Design

Objective Dyslipidemia and chronic inflammation are risk factors of cardiac fibrosis.This study was aimed to investigate their possible synergetic effects and underlying mechanisms on progression of cardiac fibrosis in apolipoprotein E knockout (ApoE-/-) mice.Methods Twenty-four ApoE-/-mice were divided into normal chow diet (control),high fat diet (HFD group),and HFD plus subcutaneously injection of 10％ casein (inflammation group) for 8 weeks.Lipid profile and serum amyloid A (SAA) were examined by clinical biochemical assays and Enzyme-Linked Immunosorbent Assay,respectively.Hematoxylin-eosin staining (HE) and Masson staining were used to evaluate the myocardial accumulation of lipid and collagen.Collagen Ⅰ protein expression was detected by immunohistochemical staining.Endothelial-to-mesenchymal transition related protein expressions were determined by Western blot.Results Serum SAA level was significantly higher in inflammation group [(127.42 ± 26.99) ng/ml] than in control [(15.40 ± 7.62) ng/ml] and HFD [(8.17 ± 0.72) ng/ml] group (all P ＜ 0.01).However serum levels of triglyceride,total cholesterol,and low density lipoprotein (LDL) cholesterol were significantly higher in HFD group than in inflammation and control groups[TG (7.53 ± 2.05) mmol/L vs.(3.43 ± 0.79) mmol/L ; TC (27.80 ± 3.99) mmoL/L vs.(14.94 ± 1.92) mmol/L ; LDL-C (11.56 ±2.56) mmol/L vs.(9.46 ± 1.31) mmol/L,all P ＜ 0.05).Foam cell formation in cardiac vessels.myocardial collagen deposit,protein expressions of collagen Ⅰ,CD31,and alpha-smooth muscle actin (α-SMA) were all significantly higher in inflammation group than in HFD group (all P ＜ 0.05) suggesting that inflammation contributes to the phenotype endothelial-to-mesenchymal transition in heart.Conclusion Inflammation exacerbates dyslipidemia mediated cardiac fibrosis in ApoE-/-mice partly through enhancing myocardial endothelial-to-mesenchymal transition.

Objective:To discuss the pathological relationship between P33ING1 and stomach cancer and its clinical significance. Methods: In 71 cases of stomach cancer specimen, twelve cases of gas tric mu cous membrane atypical hyperplasia tissues and 18 cases of normal gastric mucous membrane tissue（as control）,the expression of P33ING1 were detected b y EnVision immunohistochemical method,while the expression of P53 and Bcl -2 in stomach cancer were also detected. Results: P33IN G1 expression in mucous membrane atypical hyperplasia group and control group was positive, the expression in stomach cancer group was extremely low(62.0%，44 /71), significantly different from the other 2 groups(P<0.01).P33ING1 expression in stomach cancer was related to the tumor growth, lymph node meta s tasis and tumor polarization (P<0.01), P53 expression was related to tumor s ize, growth and lymph node metastasis (P<0.01). Bcl-2 expression was relate d to lymph node metatasis and tumor polarization.The expression of P33ING1 was related to that of P53 in stomach cancer(P<0.05),while had no relation with that of Bcl-2.Conclusion:P33ING1 may play an importa nt role in the occurrance and development of stomach cancer.It's very important to detect the expression of P33ING1 and P53 simultaneously.