Objective:To investigate the antioxidant function of Mycoplasma pneumoniae MPN662 and analyze the key active sites, and to explore the role of MPN662 in the regulation of the production of reactive oxygen species (ROS) and superoxide dismutase (SOD) in THP-1 cells. Methods:pET28a(+ )- mpn662, recombinant mutant plasmids pET28a(+ )- mpn662-Ser 66 (the 66 th Cys was mutated to Ser) and pET28a(+ )- mpn662-Ala 66 (the 66 th Cys was mutated to Ala) were constructed, recombinant proteins rMPN662, rMPN662-Ser 66 and rMPN662-Ala 66 were expressed, identified, and purified. DTNB method was employed to analyze the MetO reduction activity of rMPN662 and recombinant mutant protein. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were applied to examine the transcription level of the mpn662 gene and the expression level of MPN662 protein after Mycoplasma pneumoniae were stimulated with different concentrations of hydrogen peroxide (H 2O 2) or tert-butyl hydroperoxide (t-BHP), respectively. Fluorescent probes (DCFH-DA) and the total SOD activity detection kit were used to test the levels of intracellular ROS and SOD in THP-1 cells, which were pretreated with rMPN662, and then stimulated by Mycoplasma pneumoniae lipid-associated membrane proteins (LAMPs). Results:Mycoplasma pneumoniae rMPN662 could reduce MetO to Met, and the enzyme activities of mutant protein were significantly lower than those of rMPN662 protein. mpn662 gene mRNA transcription level and MPN662 protein expression level were significantly increased in a dose-dependent manner when Mycoplasma pneumoniae was stimulated with H 2O 2 and t-BHP. Treatment with rMPN662 before THP-1 cells were exposed to LAMPs could decrease the level of ROS and increase the production of SOD. Conclusions:Mycoplasma pneumoniae MPN662 can reduce MetO to Met, and Cys66 is the key amino acid for this activity. MPN662 can decrease the release of ROS and increase the production of SOD in Mycoplasma pneumoniae LAMPs stimulated THP-1 cells.

Objective This study aims to investigate the therapeutic effect and mechanism of Naoxinqing on non-alcohol fatty liver disease (NAFLD) induced by a high-fat diet through network pharmacology,molecular docking and in vitro and in vivo experiments. Methods ApoE-/-mice were given a high-fat diet for 12 weeks to establish the NAFLD model,followed by a 12-week Naoxinqing administration. To evaluate the therapeutic effect of Naoxinqing on NAFLD induced by a high-fat diet,biochemical and histopathological experiments were performed,including assessment of blood lipids,liver function,serum inflammatory factors,as well as Hematoxylin and eosin (HE),Oil red O,and Sirius red staining of liver. Subsequently,network pharmacology and molecular docking techniques were employed to predict the key targets of Naoxinqing. Finally,the mechanism of Naoxinqing was validated by Western Blot in HepG2 cells and liver tissue. Results The results of serum biochemistry and liver tissue pathology showed that Naoxinqing can significantly improve high-fat diet-induced hepatic lipid accumulation,hepatocellular injury,and inflammation. Network pharmacology and molecular docking analysis results suggested that Naoxinqing may affect lipid metabolism through the AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway. Finally,in vitro cell experiment confirmed that the main mechanism of Naoxinqing is to activative the AMPK/SIRT1 pathway,upregulate the expression of downstream carnitine palmitoyltransferase 1 (CPT1A),promote fatty acid oxidation,and ultimately improve NAFLD. Conclusion This study demonstrated that Naoxinqing improved NAFLD by promoting fatty acid oxidation through the activation of the AMPK/SIRT1 pathway.

The aim of this study was to establish a high performance liquid chromatography-mass spectrometry method for the determination of 5-(4′-(bromomethyl)-［1, 1′-biphenyl］-2-yl)- 1H-tetrazole(BBT1)and 5-(4′-(dibromomethyl)-［1, 1′-biphenyl］-2-yl)-1H-tetrazole(BBT2), which are two genotoxic impurities in olmesartan medoxomil. Chromatographic separation was based on an Agilent Zorbax Eclipse Plus C18(250 mm × 4. 6 mm, 5 μm)column using water(containing 0. 1% formic acid)- acetonitrile as mobile phase in gradient elution mode. Mass spectrometry was operated in positive ion mode. Selective ion monitors were set at m/z 315 for BBT1 and at m/z 395 for BBT2. Good linear correlations were observed in the range of 0. 009 4- 0. 561 0 μg/mL(r=0. 998)with the quantification limit at 9. 35 ng/mL and the detection limit at 3. 12 ng/mL for BBT1, and in the range of 0. 018 2- 0. 547 5 μg/mL(r=0. 999)with the quantification limit at 18. 25 ng/mL and the detection limit at 6. 08 ng/mL for BBT2. Furthermore, the average recoveries of the three spiked concentration level were 96. 5%(n=9, RSD=4. 8%)and 98. 0%(n=9, RSD=5. 1%)for BBT1 and BBT2, respectively. The proposed method is simple, specific and accurate, and quite suitable for the determination of BBT1 and BBT2 in olmesartan medoxomil.

Objective To compare the short-term clinical effects of totally thoracoscopic surgery and traditional open-heart sugery in the treatment of esophageal cancer. Methods 64 patients with middle thoracic esophageal cancer admitted in our hospital from June 2013 to June 2016 were selected. Among them, 32 patients underwent totally thoracoscopic surgery were identified as totally thoracoscopic group and the remaining 32 patients underwent traditional open-heart surgery were identified as control group. We compared various surgical indicators of patients in the two groups. Results In the totally thoracoscopic group, the duration of operation was(123. 8±25. 1) min, the amount of blood loss during surgery was (172. 1±30. 3) mL, the retention time of chest drainage tube was (3. 1±1. 1) d and the duration of hospitalization was (15. 6±2. 7) d. Compared with the control group, these indicators showed significant difference(P＜0. 05). The incidence of postoperative complications in the totally thoracoscopic group was 9. 3％, which was significantly lower than that in the control group(31. 3％)(P＜0. 05). The pain severity of totally thoracoscopic group reduced significantly than the control group(P＜0. 05). Conclusion Compared with the traditional open-heart surgery, totally thoracoscopic surgery for esophageal cancer has advantages of less bleeding, less trauma, few complications and less pain, which is worthy of promoting and using widely in clinic.

Objective To analyse the breast conserving surgery feasibility in patients with advanced breast cancer after neoadjuvant chemotherapy.Methods Sixty patients with advanced breast cancer were collect-ed from June 2010 to June 2011, and were divided into breast conserving surgery after neoadjuvant chemotherapy group and modified radical mastectomy group according to the intention,30 cases in each group.They were given breast conserving surgery after neoadjuvant chemotherapy and modified radical mastectomy respectively,with fol-lowing-up for 3 years.The treatment effect and contrast analysis of two groups of breast cancer CTCs positive of the two groups were compared,two groups of late follow-up of patients with local recurrence and distant metasta-sis rate,overall survival and disease free survival rate were recorded.Results Compared with the modified radi-cal mastectomy group,CTCs detection rate in breast conserving surgery group had no statistical differences(P>0.05).Two groups of patients in clinical complete remission rate,partial remission rate,disease stability factor and local recurrence and distant metastasis rate,overall survival and disease free survival rate had no statistical significances(P>0.05).Conclusion The application of breast conserving surgery after neoadjuvant chemother-apy for patients with advanced breast cancer can achieve similar effect as modified radical mastectomy surgery treatment,and can be used as a effective treatment for advanced breast cancer.

Although phosphodiesterase type 5 inhibitors (PDE5Is) are a revolution in the treatment of erectile dysfunction (ED) and have been marketed since 1998, they cannot restore pathological changes in the penis. Low-energy shock wave therapy (LESWT) has been developed for treating ED, and clinical studies have shown that LESWT has the potential to affect PDE5I non-responders with ED with few adverse effects. Animal studies have shown that LESWT significantly improves penile hemodynamics and restores pathological changes in the penis of diabetic ED animal models. Although the mechanisms remain to be investigated, recent studies have reported that LESWT could partially restore corpus cavernosum fibromuscular pathological changes, endothelial dysfunction, and peripheral neuropathy. LESWT could be a novel modality for treating ED, and particularly PDE5I non-responders with organic ED, in the near future. However, further extensive evidence-based basic and clinical studies are needed. This review intends to summarize the scientific background underlying the effect of LESWT on ED.
Animals ; Erectile Dysfunction* ; Hemodynamics ; Lithotripsy ; Male ; Models, Animal ; Penis ; Peripheral Nervous System Diseases ; Phosphodiesterase 5 Inhibitors ; Shock*

ted in a loss of cell fusion,which suggests the 928 site on G2 is crucial for cell fusion and the fusion peptide is likely on G2.

OBJECTIVETo demonstrate molecular insight into the pathology of Peyronie's disease (PD). A preliminary profile of differential gene expression between the PD plaque and control tunica albuginea was obtained with DNA microarrays. Also, to investigate the effect of intervention in PD cells, transforming growth factor-beta1 (TGF-beta1) was recruited to treat PD cell lines.

METHODSThree PD plaques and control tunica albugineas were constructed and studied. cDNA probes were prepared from RNA isolated from those cells and hybridized with the Clontech Atlas 3.6 Array. Relative changes of greater than 2.0 defined up-regulation and down-regulation, respectively. The expression of selected individual gene MCP-1 and the effect of TGF-beta1 on MCP-1 were analyzed by reverse transcriptase-polymerase chain reaction.

RESULTSSome up-regulated genes in the PD plaque detected by the Clontech assay were screened, one of them was monocyte chemotactic protein. One involved the pathogenesis of PD as a downstream gene and responded to the TGF-beta1 treatment but not CTGF. The results were also confirmed by TR-PCR in all the types of cell.

CONCLUSIONSThe cell lines from plaque tissue and normal tunica from men with PD were successfully established. The findings indicate a potential role for MCP-1 over expression in the pathogenesis of PD as a downstream gene regulated by some genes and could be a new therapeutic target in PD. The information may allow a better understanding of the basic mechanisms involved in the etiology and pathogenesis of PD. Furthermore, it may permit some strategies of therapeutic interventions combine routine methods with Chinese herbal medicine.

Cell Line ; Chemokine CCL2 ; Gene Expression ; drug effects ; Gene Expression Profiling ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Penile Induration ; drug therapy ; genetics ; pathology ; Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; pharmacology

BACKGROUNDTo investigate the relationship between immune status and rubella virus (RV) infection of central nervous system (CNS).

METHODSBALB/c mice were given dexamethaxone and cytoxan before RV JR23 strain infection. Immune functions and RV invasion to CNS were assayed at 21 days postinfection via abdominal cavity and their relationship was analyzed.

RESULTST cell functions of cytoxan group were obviously worse than those of other groups (P <0.05) by MTT method. Infection rates of dexamethaxone and cytoxan and the group without any intervention were 60%, 90% and 50% (P >0.05), respectively. Cellular immune functions of the mice with CNS infection were obviously worse than those of the mice without CNS infection (P <0.001). Specific antibodies (Ab) were assayed in all groups with ELISA and the results showed that there were no significant differences among groups (P >0.05), neither between the groups with and without CNS infections.

CONCLUSIONSRV infection of CNS may relate to cellular immune status before specific antibody was produced in the body.

Animals ; Antibody Specificity ; Central Nervous System Infections ; immunology ; virology ; Female ; Immunity, Cellular ; Male ; Mice ; Mice, Inbred BALB C ; Rubella ; immunology ; Rubella virus ; immunology ; T-Lymphocytes ; immunology

The alkaline elution and fluorometrie DNA assay were adapted to the evaluation of DNA single-strand breaks in the cultured human fetal lung 2BS cell. the DNA damage was induced in vitro by the treatment with Alternariol monomethyl ether (AME) and Alternariol (AOH) which were the main mycotoxins of Alternariol alternata. The results showed that both AME and AOH could induce the DNA single-strand breaks in the cultured human fetal lung 2BS cells and there existed the dose-response association betweenthe fractions of DNA remaining on the filters and the doses. The fractions of DNA remaining on the filters were 0.57?0.04, 0.45?0.02, 0.30?0.02, 0.18?0.01 respetively under the AME concentration of 10, 25, 50, 100?g/ml and were 0.68?0.3, 0.54?0.01, 0.47?0.03, 0.34?0.01 respetively under the AOH concentration of 0.1, 1, 5, 10?g/ml when DNA was eluted for 6hr. It were significantly different from the fractions by solvent control (0.87?0.02) (P