Objective To study the induction and detection methods of Staphylococcus aureus viable but non-culturable state (VBNC).Methods Staphylococcus aureus (ATCC13565) of 107 cfu/ml was induced by low temperature at 4 ℃,freezing at-20 ℃ and adding copper ions from 0.01 mmol/L to 0.05 mmol/L,Staphylococcus aureus was tested,and explored the resuscitation conditions by stepwise heating-heating and chemokinesis.Results Frozen 72 h at the temperature of-20 ℃ or added 0.015 mmol/L Cu2+ for 4 days of culture could induce staphylococcus aureus into the state of VBNC.Adding 0.5% tween-20 or 1% catalase for 24 h of culture could make the strain in VBNC achieve recovery.Conclusion Staphylococcus aureus can be induced into the VBNC and the recovered Staphylococcus aureus is the same as the normal bacteria in colony morphology and physiological and biochemical response in the common test medium.

Objective To identify the effects of parenteral nutrition (PN) on the gene expressions of rat intestinal inflammatory cytokincs and receptors and to explore the role of these changes in PN-related intestinal impairment. Methods Totally 12 male Sprague-Dawley rats were equally divided into the control group and the PN group. A silastic catheter was inserted into the right jugular vein of each rat. No food or water was administered to the PN group except for a continuous 24-hour PN infusion through the silastic catheter in the jugular vein. The control group, while being regularly fed, was administered with an infusion of normal saline through the silastic catheter in thc jugular vein. After 7 days, intestinal tissues were taken for electron microscopy and real-time PCR array to analyze thc microstructure change in rat intestine and thc gene expressions of inflammatory cytokines and their receptors. Results Electron microscopy revealed atrophy of microvillus, engorgement of mitochondria, cell-cell junction breakage, and several apoptotic bodies in the PN group and normal intestinal microstructure in the control group. Compared with the control group, the PN group showed an up-regulation in the gene expressions of interferon γ, interleukin-1 receptor type I , interlcukin-8 receptor type b and a down-regulation in the gene expressions of CC chemokine ligand 17 (CCL17) , CCL19, CCL21, CCL22, CCL9, CXC chemokine receptor 3, CC chemokine receptor 3 ( CCP3 ), CCR7, CCR5, C-reactive protein, and interleukin-10. Conclusions PN influences the gene expressions of rat intestinal inflammatory cytokincs and receptors. The expression of cytokine interferon γ increases and that of interleukin-10 declines, and the expressions of CCL19, CCL21, CXC chemokine receptor 3, CCR3,CCR7, and CCR5 decline. The alterations of these genes may be associated with the impairment of intestinal immune and mechanical functions.

OBJECTIVE:To assess the cost and therapeutic effects of Chinese traditional medicine and western medicine in treating varicocele sterility. METHODS:165 patients with sterility were randomly divided into 2 groups. Chinese traditional medicine group included 110 cases who took Zhang's varicosity prescription, while the western medicine group included 55 cases who took human chorionic gonadotropin, clomiphene citrate, zinc gluconate, ATP and vitamin AD,Vitamin E,Vitamin C,for 3mo～9mo normally. The therapeutic effects and adverse effects of two groups were monitored and evaluated with cost- effectiveness analysis. RESULTS:The total effective rates of Chinese traditional medicine and western medicine were 81.82%and 50.91% respectively, while the costs were 3 688.2 Yuan and 2 399.2 Yuan accordingly, C/E were evaluated to be 45.08 and 47.13 respectively and ?C/?E was 41.7. CONCLUSION:Zhang's varicosity prescription is the better choice for treatment of varicocele sterility

OBJECTIVE:To explore the clinical role of two-hour(C2)of CsA in allogeneic hematopoietic stem cell transplantation.METHODS:The whole blood CsA concentration of 9 patients of C0 and 12 of C2 were determined by FPIA method . RESULTS: The satisfactory immunosuppressive concentration was found as follow: C0 was 200～400 ?g?L-1and C2 was 500～700 ?g?L-1.Both C0 and C2 could predict the acute rejection and side-effects,but C2 is more effective.CONCLUSION:The factors that affect the whole blood concentration of CsA were complex. CsA blood concentration monitoring can be used to guide rational use of CsA,but C2 is more effective than C0.

Objective To analyze the association of blood uric acid level with the severity of coronary artery stenotic changes, metabolic syndrome (MS), and its components. Methods A total of 343 individuals ( male 223,female 120) who underwent coronary angiography and had complete data on MS and serum uric acid were collected. The severity of coronary artery disease (CAD) was assessed by the coronary stenesis index (CSI). MS was diagnosed according to the Guideline on Prevention and Treatment of Blood Lipid Abnormality in Chinese Adults. Results (1)The mean uric acid level was significantly lower in women than in men [ ( 306.3±76.9 vs 358.9±85.2 ) μmol/L, P＜0.01 ]. The prevalence of MS and its components showed no difference between men and women. (2) The uric acid level in women with 3 components was higher than those with1( P＜0. 01 ) or 2 ( P＜0.05 ) components of metabolic disorders, but not in men. (3) Quartiles of concentration of uric acid were computed. Compared with those in the lowest quartile of uric acid, women in the highest quartile had higher CSI score [ 7.0 (2.5-12.0) vs 2. 0( 0.0-6.0), P= 0. 025 ]. Moreover, the uric acid level was higher in women with multivessel lesions than nonCAD patients [ (327.0±81.9 vs 284.9±78.6) μmol/L, P = 0.033 ]. However, no correlation was found between uric acid level and the severity of coronary artery lesion in men. (4) Logistic regression showed that age (β=0.042, P=0. 007) and dyslipidemia(β=0.836, P=0. 037 ) were the independent risk factors of CAD in men, and hypertension(β=1. 127, P=0.039) and dyslipidemia(β=0.901, P=0.009)in women. Conclusions In women with higher uric acid level, the clustering of metabolic abnormalities was increased, and the coronary artery lesion was more severe. High uric acid level might be a marker of CAD for women.

Objective To analyze the clinical characteristics of coronary artery disease (CAD) in patients with nonalcoholic fatty liver disease ( NAFLD). Methods Totally 234 subjects underwent coronary angiography, including 148 men and 86 women with complete data on metabolic syndrome ( MS) and abdominal ultrasonography; the mean age was 66. 6 years. The severity of CAD was assessed by coronary stenosis index (CSI). Metabolic syndrome was diagnosed according to the Guideline on Prevention and Treatment of Blood lipid Abnormality in Chinese Adults. Results Ultrasonography revealed that 62 patients had NAFLD (26. 5%). In patients with NAFLD, the prevalence of central obesity was higher than those without NAFLD (75. 8% vs 50. 0% , P< 0.01). With regards to age, CAD patients with NAFLD were more common in patients under 60 years (27.4% vs 13.7% , P=0.005). CSI score was similar in CAD subgroup and CAD & NAFLD subgroup (P>0.05), however the age of patients in CAD & NAFLD subgroup was significantly lower compared to CAD subgroup ( P = 0.006). According to the results of logistic regression, central obesity was the independent risk factor of NAFLD (β= 1.701, P<0.001). Logistic regression demonstrated that age was independently associated with CAD (β = 0.032, P=0.027). Further more, multiple stepwise regression analysis showed that age was the single parameter that best predicted CSI score (β= 0. 125, P = 0. 022). Conclusions It is important to screen coronary artery disease in middle aged patients with central obesity or NAFLD.

Objective To investigate the correlation of multidrug resistance gene 1 (MDR1),NR3C1 gene polymorphisms and clinical risk factors with efficacy,dependence,and resistance of glucocorticoid (GC) in patients with inflammatory bowel disease (IBD).Methods Anti coagulation blood samples of 196 healthy controls and 105 IBD patients received GC therapy were collected.There were 62 ulcerative colitis (UC) and 43 Crohn's disease (CD) in the IBD patients.The number of GC sensitive,GC dependent and GC resistant of UC patients were 36,13 and 13,respectively,and those of CD patients were 24,11 and eight.GC refractoriness included GC dependence and resistance.The genotype of MDR1 C3435T and NR3C1 Bcl Ⅰ of all the subjects was detected by the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR).The correlation between each genotype frequency,clinical features of patients with IBD and the efficacy of GC treatment was analyzed by Chisquare test,Fisher exact probability method or t test.Results Among UC patients,the disease course of GC refractory group and GC resistant group was longer than that of GC sensitive group ((6.660±1.523)years,(6.500±1.111) yearsvs (3.350±0.697) years,t=2.211,P=0.031; t=2.930,P=0.005).The serum level of C reaction protein (CRP) of GC refractory group was higher than that of GC sensitive group ((47.628±13.913) mg/Lvs (16.854±4.121) mg/L,t=2.121,P=0.047).The chronic relapse type was more common in GC refractory UC patients (Fisher exact probability method,P=0.035),and severe patients were more common in UC with GC resistance (Fisher exact probability method,P=0.021).The white blood cell count of GC resistant and GC refractory CD patient was lower than that of GC sensitive CD patients ((5.710 ± 0.604) ×109/L,(5.878±0.405) × 109/L vs (7.814 ±0.670) × 109/L,t=2.334,P=0.028; t=2.045,P=0.018).Patients with extraqntestinal manifestations was more common in CD with GC resistance (Fisher exact probability method,P=0.035).There was no statistically significant difference in the frequencies of MDR1 C3435T,NR3C1 Bcl Ⅰ genotypes,allelic genes and gene carrier among control group and GC sensitive dependent and resistant group of IBD patients.However,the frequency of MDR1 C3435T gene carrier was significantly different between GC sensitive group and GC refractory group,especially between GC sensitive group and GC resistance group (68.33％ vs 48.89％,x2 =4.051,P=0.044; 68.33％ vs 42.86％,x2 =4.274,P =0.039).Conclusions GC sensitivity of IBD patients with MDR1 C3435T loci T gene carrier was higher than that of IBD patients without T gene carrier.NR3C1 gene polymorphisms was not related with GC resistance and GC dependence.Compared with GC sensitive IBD patients,in GC resistant and GC dependent IBD pantient UC patients with long disease course,chronic relapse type,severe type,high level of CRP and CD patients with low white blood cell count and extra-intestinal manifestations were more common.

Objective To investigate the changes in peripheral blood 25-hydroxyvitamin D3 [25-(OH)D3],CD4+ regulatory T (Treg) cells,and Th17 cells in patients with primary biliary cirrhosis (PBC) and their mechanism of action in PBC.Methods A total of 22 patients with PBC were enrolled and the male/female ratio was 1∶21,with a mean age of 61± 12 years.There were 7 healthy volunteers matched for age in the normal control group.Electrochemiluminescence immunoassay was used to measure the peripheral blood 25-(OH)D3 level in the PBC group and normal control group,and flow cytometry was used to analyze the changes in Th1 7 cells and CD4+ Treg cells.The t-test,rank sum test,Pearson correlation analysis,or Spearman's rank correlation analysis was used for statistical analysis according to the type of the data.Results The PBC group had a significantly lower serum 25-(OH)D3 levcl than the normal control group (9.49±3.65 vs 27.35±2.35 ng/ml,P ＜ 0.01).Compared with the normal control group,the PBC group had a significantly higher percentage of Th1 7 cells (2.05％±1.17％ vs 0.99％±0.12％,P ＜ 0.01) and a significantly lower percentage of CD4+ Treg cells (2.54％± 1.14％ vs 3.78％±0.51％,P ＜ 0.05);there was a significant difference in Th17/Treg ratio between the PBC group and the normal control group (1.00±0.63 vs 0.26±0.02,P ＜ 0.01).In the PBC group,peripheral blood 25-(OH)D3 was not correlated with Th17 cells or Th17/Treg ratio (r =-0.062 and-0.328,P ＞ 0.05),while it was positively correlated with the percentage ofCD4+ Treg cells (r =0.468,P ＜ 0.05).Conclusion Patients with PBC have significant reductions in peripheral blood 25-(OH)D3 and percentage of CD4+ Treg cells,a significant increase in the percentage ofTh17 cells,and immune unbalance of Th17 cells and CD4+ Treg cells.25-(OH)D3 can upregulate the percentage of CD4+ Treg cells and thus affect the development and progression of PBC,and exogenous vitamin D may improve immune function in PBC patients.

Objective To investigate the outcome of postoperative hypopituitarism and hormone replacement in patients with pituitary adenoma,and to analyze the potential factors related to postoperative hypopituitarism.Methods A total of 215 postoperative patients with pituitary adenoma were analyzed.Pituitary functions( including gonadal,thyroid,and adrenal axes ) were asessed by strict criteria.Data of surgery history and hormone replacement situation were collected for statistical analysis.Results The prevalence of hypopituitarism was 54.0％,including 36.7％ hypogonadism,32.6％ hypothyroidism,and 28.4％ hypoadrenalism.Replacements of gonadal steroid,glucocorticoid,and thyroxine were carried out in 25.6％,84.3％,and 80.6％ of the cases,respectively.Univariate analysis showed that male sex and large tumor were related to hypopituitarism. Conclusion After pituitary adenomectomy,approximately half of the patients present anterior pituitary dysfunction,while quite a number of them have not been treated appropriately.

We studied the effect of surfactin on cell proliferation, apoptosis and the cytoskeleton in human breast cancer cell line MCF-7 in vitro. The result of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) showed that the surfactin inhibited proliferation of MCF-7 cells in a dose- and time-dependent manner, with IC50 at 48 h of 27.3 micromol/L. Surfactin-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological changes by AO/EB staining. Flow cytometric analysis also demonstrated that surfactin caused time-dependent apoptosis of MCF-7 cells through cell arrest at G2/M phase. Immunofluorescence and Western blotting showed that surfactin significantly suppressed the expression of vimentin, induced the alpha-tubulin depolymerization and rearrangement and then the skeleton system of the cells changed dramatically. Based on our findings, surfactin can significantly inhibit the growth of MCF-7 cells and induce apoptosis.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytoskeleton ; drug effects ; Female ; Humans ; Lipopeptides ; pharmacology ; Peptides, Cyclic ; pharmacology