Humans ; Infant, Newborn ; Syphilis Serodiagnosis ; Syphilis, Congenital ; diagnosis

Objective To investigate the role of cathepsin G in photoaged fibroblasts. Methods Human fibroblasts were cultured and induced to premature senescence using UVA + MOP methods. Senescence-associated-β-galactosidase (SA-β-gal) stain was used to evaluate the positive rate of aged cells. The mRNA and protein expression of cathepsin G in photoaged fibroblasts were detected by real-time RT-PCR and Western blot techniques. Results Over 98 % induced cells presented a positive SA-β-gal straining. The expression of cathepsin G, detected by Western blot, was increased to (1. 70±0. 028) times of the control. And RT-PCR revealed that the synthesis of cathepsin G mRNA was also up-regulated to 1. 42±0. 09. Conclusion The results of our study demonstrates a significant correlation between photoaged fibroblasts and cathepsin G. The up-regulation of cathepsin G may play an important role in the damages of extracellular matrix and activation of MMPS in photoaged human skin.

Objective To observe the short-term effect of Paris polyphylla Smith var. yunnanensis powder combined with anti-tuberculosis drugs in the observation of superficial lymph node tuberculosis. Methods A total of 170 patients were randomly divided into two groups: 80 in the control group and 90 in the observation group. The observation group consisted of nodular type, infiltrating type and abscess type, each of which had 30 cases. The control group were treated with Isoniazid, Rifampicin, Pyrazinamide and Ethambutol. Besides the four medications, the observation group were treated with external application of Paris polyphylla Smith var. yunnanensis. Results The response rate was 30.00％in the control group and 64.44％in the observation group. The response rate in the observation group was higher than that in the control group, with a statistically significant difference ( <0.01). In the observation group, the response rates of abscess, infiltration and nodule were 76.67％, 73.33％and 43.33％, respectively.By the comparative analysis, the response rate of infiltration was higher than that of nodule ( <0.05), with a significant difference; the response rate of abscess was also higher than that of nodule ( <0.05), with a significant difference. There was no significant difference between the response rates of abscess and infiltration ( >0.5) .Conclusions The external application of Paris polyphylla Smith var. yunnanensis powder combined with anti-tuberculosis drugs is curative in the observation of superficial lymph node tuberculosis, especially in the types ofinfiltrating and abscess.

Objective To construct a risk assessment tool framework for surgical wound infection(SWI). Methods The assessment system was constructed by referring to expert interviews, relevant domestic and foreign literatures, and the latest SW1 guidelines. After two rounds of Delphi expert consultation, the assessment items were drawn up, selected and modified, and the initial scale was established. Results The positive coefficients of two rounds of expert Delphi methods were 84％ and 100％ respectively; expert personal authority coefficient were both ＞75％, the all expert authority coefficient was 0.86; two rounds of Kendall's W coefficients were 0. 450 and 0. 441 respectively (all P く0. 05). The recommendations of two rounds of experts were 26 and 5 items respectively. The final assessment system indexes included 4 first-level indicators, 10 second-level indicators, and 27 third-level indicators. Conclusion The risk assessment tool framework for SWI is preliminarily established, which can provide a scientific basis for the effective evaluation of the risk of postoperative SWI.

OBJECTIVETo investigate the effects and clinical pregnancy outcomes of intracytoplasmic sperm insemination (ICSI) with microamount frozen-thawed sperm obtained by percutaneous epididymal sperm aspiration (PESA) or testicular sperm aspiration (TESA) in azoospermia patients.

METHODSWe divided 365 azoospermia patients treated by ICSI into an experimental group (n = 123) and a control group (n = 242) , the former with microamount frozen-thawed sperm, and the latter fresh sperm obtained by PESA or TESA. The rates of fertilization, good embryos, clinical pregnancy, miscarriage, ectopic pregnancy and multiple pregnancy were analyzed and compared between the two groups.

RESULTSWith PESA, the experimental group showed no statistically significant differences from the control group in the rates of fertilization (75.67% vs 76.49%), good embryos (64.96% vs 66.09%), clinical pregnancy (55.21% vs 57.22%), clinical miscarriage (13.21% vs 12.61%), ectopic pregnancy (3. 77% vs 5.41%) and multiple pregnancy (37.74% vs 37.84%) (P > 0.05); nor with TESA (74.41% vs 76.43%, 64.63% vs 66.35%, 46.81% vs 53.39%, 18.18% vs 14.55%, 4.55% vs 1.82%, 37.74% vs 37.84%, P > 0.05). The revival rate of the frozen-thawed sperm from PESA was 70.07%, not significantly different from that of TESA (62.67%) (P > 0.05).

CONCLUSIONICSI with frozen-thawed micro-amount sperm obtained by PESA or TESA is a safe, economic and effective method for the treatment of azoospermia. The techniques for reviving frozen sperm from PESA or TESA remain to be optimized, and whether these techniques may result in long-term genetic risks in the offspring deserves further investigation.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; methods ; Sperm Retrieval

OBJECTIVETo investigate effect of tumor necrosis factor-alpha (TNF-alpha) on the Src-suppressed C kinase substrate (SSeCKS) in C6 glioma cells.

METHODSCultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-alpha (2 ng/mL) for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-alpha (0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL) for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS's subcellular localization.

RESULTSTNF-alpha induced rapid phosphorylations of protein kinase C (PKC) substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-alpha induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220.

CONCLUSIONTNF-alpha activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.

A Kinase Anchor Proteins ; Animals ; Astrocytes ; metabolism ; Brain Neoplasms ; metabolism ; Cell Cycle Proteins ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation ; Glioma ; metabolism ; Immunohistochemistry ; Protein Kinase C ; metabolism ; Protein Transport ; physiology ; Random Allocation ; Rats ; Signal Transduction ; physiology ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; administration & dosage ; physiology

OBJECTIVETo study the survival rate and secretory function of islet cells in rats under condition of three-dimensional microgravity.

METHODSIsolated islet cells were assigned to flask-cultured or bioreactor-cultured. Survival rate of islets cultured for days 3, 7, 14 in stationary flasks or microgravity bioreactors was measured by AO-PI double-staining. Cultured islets were identified by dithizone (DTZ) staining, and insulin contents of different culture liquids were measured by radioimmunoassay.

RESULTSPancreatic islets stained nacarat with DTZ were easily visualized. When islet cells were cultured for 7 days and 14 days, survival rate of bioreactor-cultured islets was (0.9000 +/- 0.0107)% and 0.8038% +/- 0.0092% and higher than flask-cultured islets (P < 0.01). Insulin level of bioreactor-cultured islets is (70.875 +/- 0.31) m micro /L on the cultured 7 days while flask-cultured islets is (41.246 +/- 0.35) m micro /L. There was statistically significant difference of insulin production between the two groups (P < 0.01). Bioreactor-cultured islet contents were higher than flask-cultured on the cultured 14, 21 and 30 days (P < 0.01).

CONCLUSIONSIslet cells survival rate and secretory function revealed that bioreactor-cultured islets functioned better compared with flask-cultured islets.

Animals ; Bioreactors ; Cell Culture Techniques ; methods ; Cell Survival ; Cells, Cultured ; Insulin ; secretion ; Islets of Langerhans ; cytology ; secretion ; Male ; Rats ; Rats, Wistar ; Weightlessness Simulation

OBJECTIVETo investigate the expression and correlation of Skp2 and p27kipl in non-Hodgkin's lymphoma.

METHODSThe expression of Skp2, p27(kip1) and Ki-67 (the proliferation index)were detected in sections of 92 cases of non-Hodgkin's lymphoma (NHL) and 14 cases of reactive lymph nodes by immunohistochemistry and histopathology. The expression of Skp2 and p27(kip1) in 4 NHL cell lines were detected by Western blot.

RESULTSThe expression of Skp2 in NHL cases were significantly higher than that in reactive lymph nodes (except the germinal centers), positively correlated with proliferation activity, and an increasing tumor aggressiveness was associated with the increased expression of Skp2. The expression of p27(kip1) protein in NHL cases were significantly lower than that in reactive lymph nodes (except the germinal centers), negatively correlated with proliferation activity, and an increasing tumor aggressiveness was associated with decreased expression of p27(kip1). The statistical analysis indicated that there was no obvious correlation between Skp2 and p27(kip1) expression in NHL tissues.

CONCLUSIONThe higher expression of Skp2 and lower expression of p27(kip1) in NHL tissues may play a role in the tumorigenesis and development of NHL.

Blotting, Western ; Castleman Disease ; metabolism ; pathology ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; Cytoplasm ; metabolism ; Humans ; Immunohistochemistry ; Intracellular Signaling Peptides and Proteins ; metabolism ; Ki-67 Antigen ; metabolism ; Lymph Nodes ; metabolism ; pathology ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; metabolism ; pathology ; S-Phase Kinase-Associated Proteins ; metabolism

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae

Objective To evaluate the AChE and nAChR in the NMJ and the morphology of the muscle in the bilateral triceps surae after the unilateral shock wave therapy. Method 60 male New Zealand rabbits weighing (2± 0.2) Kg were used in this study. Two thousand shock waves at an energy flux density of 1.5bar and the frequency of 10Hz were applied to their left calf muscles. Divided into six groups, both sides of the triceps muscle of calf were taken out on the day of the shock wave and the1,2,4,6 and 8 weeks after the treatment. HE staining was used to observe the morphological changes of muscle tissue and the average optical density was measured after AChE stain so as to calculate the receptor count after Acetylcholine receptor immunohistochemistry. Result No abnormal morphological abnormalities were observed in all rabbits. In the first five groups, the AChE was significantly higher in the side of the shockwave treatment compairing with the control side (<0.05),slow decrease after 1 week after the treatment. In the first five groups, the nAChR was significantly lower in the side of the shock wave treatment compairing with the control side ( <0.05), and gradually increased to normal after 8 weeks. Conslusion Suitable dose of shock wave will not have a greater impact on morphology of muscle tissue. After the shock wave treatment, the amount and degree of stimulate of muscle cells were decreased, and the production of action potentials was reduced. While the experimental side AchE and AchR in shock wave treatment day to 8 weeks after treatment showed a significant trend to normal, it shows that the effect of shock wave on NMJ is transient and reversible.