Objective· To investigate whether vitamin E could reverse the disruptive effects of bisphenol A (BPA) on steroidogenesis and to explore the optimal vitamin E concentration.Methods· Rat primary granulosa cells were extracted and exposed to BPA (0,0.01,0.1,1,10,100 μmol/L).After 48 h of incubation,culture medium was collected and estradiol (E2) and progesterone (P4) were measured using ELISA kits.Then,granulosa cells were incubated with 5 μmol/L(average concentration in follicular fluid) or 25 μmol/L (high concentration in follicular fluid) vitamin E (α-tocopherol) or vitamin E (5 μmol/Lor 25 μmol/L) plus BPA (10 μmol/L or 100 μmol/L) for 48h,E2 and P4 were measured.Results· BPA at 10 μmol/L and 100 μmol/L suppressed E2 and P4 production in a dose-dependent manner (P＜0.05).Vitamin E at 25 μmol/L significantly increased E2 and P4 levels by (44.89±15.18) ％ and (43.33±8.82) ％(P＜0.05),respectively.Coincubation of the granulosa cells with BPA and vitamin E (5 μmol/L or 25 μmol/L) restored the productions of E2 and P4,which were not significantly different from the control group (P＞0.05).Conclusion· Vitamin E (5 μmol/L/25 μmol/L) could reverse BPA-induced reduction in steroid hormone production in rat ovary granulosa cells,and the antagonistic effect of 25 μmol/L vitamin E was more obvious than that of 5 μmol/L vitamin E.

Objective:To observe therapeutic effect of lyophilized recombinant human brain natriuretic peptide (Lrh-BNP)on stress cardiomyopathy (SCM)complicated acute bump failure.Methods:A total of 23 patients with SCM complicated acute bump failure were randomly divided into routine treatment group (n=10,received routine treatment)and Lrh-BNP group (n=13,received Lrh-BNP based on routine treatment).Clinical symptoms and signs,cardiac function :left ventricular ejection fraction (LVEF),stroke volume (SV),cardiac index (CI),peak early diastolic velocity/peak late diastolic velocity (E/A)assessed by echocardiography before and after treatment, and total effective rate were compared between two group.Results:Total effective rate of Lrh-BNP group was sig- nificantly higher than that of routine treatment group (92.3% vs.80.0% ,P<0.05).There were no significant difference in cardiac function indexes between two groups before treatment (P>0.05 all);after treatment,com- pared with routine treatment group,there were significant rise in LVEF [(50.2±16.3)% vs.(59.4±14.1)%],SV [(39.5±10.4)ml vs.(48.3±12.5)ml],CI [(3.7±1.1)L min-1 m-2 vs.(4.6±1.4)L min-1 m-2 ]and E/A [(1.0±0.5)vs.(1.3±0.7)]in Lrh-BNP group,P<0.05 all.Conclusion:Lrh-BNP possesses significant thera- peutic effect on stress cardiomyopathy complicated acute left heart failure.

Macrolactins are 24-membered macrolides produced by unidentified marine bacterium, Actinomadura sp. and Bacillus sp., which exhibit both antibacterial and antitumor activities in vitro. The environmental strain X-2 which was isolated from the sediment of the East China Sea produce Macrolatin A, B and O. In this study, a set of degenerate oligonucleotide primers, designed for amplification ketosynthase(KS) domains, had been employed to identify KS gene fragments of the X-2 DNA samples. One 645 bp KS fragment(GenBank accession no. EF486351)had been cloned and used as a probe to screen the genome DNA fosmid library of X-2. Three positive clones were selected and sequenced, Homologous analysis and the function prediction of the obtained PKS gene fragments suggested that macrolactin is the Polyketide Biosynthesis Product of the gene cluster obtained in the environmental strain X-2.

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.

Objective To evaluate the effect of hydrogen-rich saline on Toll-like receptor 4 (TLR4) /nuclear factor kappa B (NF-κB) signaling pathway in the sciatic nerve of rats with diabetic neuropathic pain (DNP).Methods Pathogen-free male Sprague-Dawley rats, aged 8 weeks, weighing 180-210 g, were used in the study.DPN model was established by intraperitoneal injection of 1％ streptozocin (STZ) 65 mg/kg.Twenty-four diabetic rats were randomly divided into 2 groups (n =12 each) using a random number table: DPN group and hydrogen-rich saline group (HRS group).Another 12 normal rats were randomly selected and served as control group (group C).At 14 days after STZ injection, hydrogenrich saline 5 ml/kg was injected intraperitoneally once a day for 14 consecutive days in group HRS, while the equal volume of normal saline was given in C and DNP groups.Mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 days before STZ injection (T0) , and 7, 14, 21 and 28 days after STZ injection (T1-4).The motor nerve conduction velocity (MNCV) of the right hindlimb was measured after pain threshold was measured at T4.After measurement of neurological function was completed, the expression of TLR4 and NF-κB was detected in the sciatic nerve (by Western blot) , the tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) contents in sciatic nerves were measured by enzyme-linked immunosorbent assay, and the neuronal apoptosis was detected by TUNEL.The apoptosis index was calculated.Results Compared with group C, the MWT was significantly decreased at T1-4, TWL was shortened at T2-4, and MNCV was decreased at T4, the expression of TLR4 and NF-κB, contents of TNF-α and IL-6, and apoptosis index were increased in HRS and DNP groups (P＜0.05).Compared with group DNP, the MWT was significantly increased, and TWL was prolonged at T3,4 MNCV was increased T4, and the expression of TLR4 and NF-κB, contents of TNF-α and IL-6, and apoptosis index were decreased in group HRS (P＜ 0.05).Conclusion Hydrogen-rich saline can mitigate DNP through blocking TLR4/NF-κB signaling pathway in the sciatic nerve of rats.

Objective To compare the safety , surgery effects , pain scores of the close-distance-two-port video-assisted lobectomy and systematic dissection of the mediastinal nodes , and single-port video-assisted thora-coscopic surgery (VATS). Methods Between October 2012 and January 2015 in Peking University First Hospi-tal and Beijing Hospital , 269 patients who were going to be performed lobectomy and systematic lymph node dissection, were gathered and 205 patients were included in this study finally. Of the total, 122 patients were performed close-distance-two-port VATS , and 83 patients underwent single-port VATS. The clinical data were gathered, and statistically analyzed. Result In both groups, no severe postoperative complications or death oc-curred. No significant differences existed between the two groups in terms of intraoperative blood loss , duration and volume of chest tube drainage , postoperative pain score , hospital stay after surgery and hospitalization ex-penses (P > 0.05) but the data about operating time, number of dissected lymph nodes, complications(subcuta-neous emphysema , air-leak from the port of drainage tube and poor wound healing ) were significantly different (P < 0.05). Conclusions Compared with the single-port VATS, the method of close-distance-two-port video-as-sistant lobectomy and systematic dissection of the mediastinal nodes is safe and practicable with definite thera-peutic effect, less operation difficulty and complications.

Objective To examine the sleep status and relevant psychosocial factors in patients with colorectal cancer before surgery and improve their sleep quality. Methods A cross?sectional survey method was used. Participants were 107 cases of patients with colorectal cancer from the Department of Anorectal Surgery in The First Hospital ,which is affiliated with China Medical University. The Athens insomnia scale(AIS),Ham?ilton depression scale(HAMD),Hamilton anxiety scale(HAMA),perceived social support scale(PSSS),self?esteem scale(SES),medical cop?ing questionnaire(MCMQ),memorial University of Newfoundland scale of happiness(MUNSH),Eysenck personality questionnaire(EPQ),and Wong?Baker face scale were used to assess preoperative sleep in patients with colorectal cancer and related psychosocial factors. Results Among the 107 cases,there were 24 cases of insomnia,with an insomnia incidence of 22.43%. No statistical differences were found in demographic charac?teristics and clinical characteristics(P>0.05). According to the degree of insomnia,there were significant differences between groups(P<0.05) for depression,anxiety,social support,avoidance,yield,happiness,and EPQ?N. Before the surgery,the degree of depression,anxiety,and EPQ?P had a significant positive effect on the degree of insomnia (P< 0.05). Conclusion The degree of insomnia before surgery in patients with colorectal cancer is closely associated with depression,anxiety,coping styles,social support,and personality characteristics.

OBJECTIVETo investigate the effects of substance P on cultured rat osteoclasts.

METHODSNeurokinin-1 (NK1) receptor expression in osteoclasts was examined by immunohitochemical method, and changes of bone resorption activity caused by substance P and NK1 receptor antagonists were detected by pit formation assay.

RESULTSImmunoreactivity for NK1 receptor was distributed in the cytoplasm of osteoclasts. The average of pit formation areas significantly increased with addition of substance P (10(-7)-10(-4) mol/L) (P < 0.05), but the number of pitformations did not change (P > 0.05). NK1 receptor antagonists inhibited the enhancement of the bone resorption by substance P addition.

CONCLUSIONThe findings suggested that substance P may stimulate osteoclasts and result in bone resorption by the mediation of NK1 receptor.

Animals ; Osteoclasts ; Rats ; Receptors, Neurokinin-1 ; Substance P

ABSTRACT:The virulence genes ,hemolytic and antibiotic resistance characteristics from 19 isolated Streptococcus suis se‐rotype 2 in Anhui Province were investigated in this study .The PCR method was developed for detecting cps2J ,mrp ,ep f and sly gene;the plate method and micro‐ELISA were developed for detecting the hemolytic type and hemolytic titer ;the K‐B method was developed for detecting antibiotic resistance of 25 kinds of antibiotics .Results showed that there were 11 strains with the dominant virulence genotype of cps2J+ /mrp+ /ep f+ /sly+ accounting for 57 .9% .All strains were α‐hemolysis orβ‐hemolysis ,with hemolytic titer of 1∶4 to 1∶128 .Strains sensitive to rifapentine ,ceftazidime ,florfenicol and cefazolin was existed in high frequency with the sensitive rates for 84 .2% ,and the resistance to doxycycline ,tetracycline ,bacitracin and sul‐famethoxazole was existed in high frequency with the resistance rates for 82 .9% .The multi‐antibiogram typing was doxycyc‐line ,tetracycline ,bacitracin and sulfamethoxazole account for 63 .2% .In conclusion ,the distribution of virulence genes of S .suis 2 Anhui isolates was similar with that of domestic reported strains ,and there was some differences with that of over‐seas reported strains .CPS ,MRP ,EPF and SLY are important virulence factors and the incompleteness of sly gene had no effect to its haemolyticus .The multi‐antibiotic resistance of S .suis 2 Anhui isolates was serious .There is difference for anti‐biogram typing between S .suis 2 Anhui isolates and those in other areas .

OBJECTIVETo explore the mechanism governing the reversal of multidrug resistance in human breast carcinoma cells by chelerythrine.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to determine the expressions of protein kinase Cα (PKCα) and multidrug resistance-related genes ABCG2, ABCC1, MDR1, and P-glycoprotein (P-gp) in MCF-7Taxol cells after treatment with chelerythrine and phorbol-12-myristate-13-acetate (PMA). Also, the antitumor effect of PMA or chelerythrine and effects of PKCα activator or inhibitor in combination with paclitaxel or adriamycin on multidrug resistance in MCF-7Taxol cells were evaluated by MTT.

RESULTSRT-PCR or Western blot showed that the expressions of MDR1 and P-gp were significantly higher in MCF-7Taxol cells exposed to PMA stimuli (both P0.05).

CONCLUSIONPKCα inhibitor chelerythrine can reverse multidrug resistance in breast carcinoma cells by inhibiting the expressions of MDR1 and P-gp expression in vitro.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Benzophenanthridines ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Humans