Objective To investigate the effect of castration on heart function in male rabbits postinfarction congestive heart failure model. Methods 32 male 4-5 months old New Zealand rabbits were randomly assigned to ligation of first branch of left coronary artery or sham operation (sham group, n=8). 3 weeks later 20 survived MI model rabbits were randomly subjected to castration (castration group, n=10) or sham operation (MI group, n=10). Serum FT, DHEAS levels and LVEF were checked at week 0, 3 and 15. Heart diameter and left ventricular wall thickness were measured at week 15. Results The serum FT and DHEAS levels decreased significantly at week 3. In MI group and castration group serum androgen level was lower and heart weight, ventricular diameters, ventricular wall thickness corrected by body weight were higher than that in sham group at week 15. The androgen level and body weight of castration group were lower than that in MI group. But LVEF in castration group was not higher than MI group. Conclusion There was no benefit effect of castration on heart function in male rabbits postinfarction congestive heart failure model.

ObjectiveTo investigate the effects of low dose testosterone undecanonate (TU) and dehydroepiandrosterone (DHEA) on serum androgen levels in male adult rabbits postinfarction congestive heart failure (CHF) model.Methods48 male 4～5 months old New Zealand rabbits were randomly assigned to ligation of first branch of left coronary artery (n=40) or sham operation (n=8). 3 weeks later, 30 survived model rabbits were randomly treated with TU 3 mg/kg per 2 weeks intramuscular injection (TU group, n=10), DHEA 3 mg/kg/d oral (DHEA group, n=10) or placebo (placebo group, n=10) for 3 months. Serum androgen changes were recorded at 0th, 3rd and 15th week.ResultsAt 3rd week after peration, left ventricular ejection fraction of model animals was 47.6±4.5% lower than that of the sham group (66.7±5.8%) (P<0.05). Serum free testosterone level of model animals returned to normal ranges after low dose TU or DHEA treatment, otherwise DHEAS level was higher than the sham group (P<0.05).ConclusionThe doses used in this research reached physiological dosage to male adult rabbits in postinfarction CHF model.

ObjectiveTo investigate the gene expressions of TAZ and Wnt/β-catenin on the postosteogenic cells of mesenchymal stem cells(MSC)in multiple myeloma(MM)patients and to explore the potential therapeutic target of multiple myeloma bone disease (MBD).MethodsBone marrow mononuclear cells MNC from MM and controls were isolated,cultured,expanded and then induced to osteogenic differentiation.Realtime quantitative RT-PCR was employed to detect the osteogenic markers (TAZ,Wnt/β-catenin,OPN,OC,ALP and Cbf α1); and alizarin red staining for mineral deposition.The mRNA expressions of TAZ and Wnt/β-catenin in the two groups were analysed.ResultsAlizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC.The mRNA expressions of OC,ALP and Cbf α1 were 2.0958±0.5665,2.6670±0.3847,0.8463±0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly higher than those of pre-osteogenic cells(1.3487±0.9291,1.1452±0.6054,0.4439±0.2945) (t =2.171,6.709,2.795; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP and Cbf α1 were 2.1096±0.8267,2.8991±0.3531,4.3045±0.2844,1.3273±0.4075,respectively,on the post-osteogenic cells of MSC in the controls,which were significantly higher than those of pre-osteogenic cells (1.2200±0.9091,0.8780±0.3927,1.9161±0.2684,0.6736±0.2513) (t =2.289,12.103,25.134,4.411; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP,Cbf α1 were 1.2710±0.5636,2.0958±0.5665,2.6670± 0.3847,0.8463+0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly lower than those of control groups(2.1096 ±0.8267,2.8991 ±0.3531,4.3045±0.2844,1.3273±0.4075) (t =-2.650,-3.805,-10.822,-2.841; all P ＜ 0.05).The mRNA expression of TAZ and β-catenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic cells of MSC in MM patients,which were significantly lower than those of control groups (4.4140±0.8325,0.9516±0.2920) (t =±5.085,-3.235;both P ＜ 0.05).ConclusionThe gene expressions of OPN,OC,ALP and Cbf α1,the osteogenesis related genes,are increased in post-osteogenic cells of MSC,which showed the MSC have been successfully induced to osteoblasts.Comparing with control groups,the osteogenic potential of MSC in MM patients is lower.Based on the above research,TAZ and Wnt/β-catenin may present a novel target for the future therapy of MBD.

Objective To construct a recombinant adenovirus vector of Hairpin RNA specific for MRP1 gene and study its inhibition of MRP1 gene expression in K562/AS2 cell line resistant to AS_2O_3 (ATO). Methods A MRP1-specific hairpin RNA recombinant adenovirus vector was constructed and used to infected K562/AS2 cells. Expression level of MRP1 mRNA detected by real-time fluorescent quantitative PCR. MRP1 protein detected by flow cytometry. MTT method was used to detected the cytotoxicity of ATO and etoposide. Results MRP1 mRNA and protein expression level in K562/AS2 cells before and after the pAd-MRPl-shRNA adenovirus infection was (34.70±0.28 vs 4.19±0.03, P <0.05) and (26.40±0.16 vs 10.85±0.37, P<0.05), respectively. RR of K562/AS2 to arsenic trioxide and etoposide was (11.4078±0.3183 fold vs 1.6126±0.3015 fold, P<0.05) and (5.9141 ±0.0149 fold vs 1.7664±0.1038 fold, P <0.05), respectively. The reversal fold of ATO and etoposide was (7.2409±1.3668) and (3.3555±0.1886), respectively. Conclusion Successfully constructed pAd-EGFP-U6-shRNA-MRPl adenovirus vector, the vector of infection K562/SA2 cells can inhibit MRP1 gene expression and reverse the resistance of the ATO and etoposide.

OBJECTIVETo study the possible pathogenic mechanism of liver injury in type 2 diabetes mellitus (T2DM) and the intervening effect of puerarin on it.

METHODSMice with T2DM (KKAy) were randomly divided into two groups, the model group and the puerarin group. And the C57BL/J mice of the same age were set up as normal controls. They were sacrificed at 28 weeks old for observing serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST) by automatic biochemistry; liver cell apoptosis by flow cytometry; pathomorphology by electron microscope; and mRNA expressions of bcl-2 and bax genes by RT-PCR; as well as the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Na(+)-K(+)-ATPase; and content of malondialdehyde (MDA) in liver tissue by spectrophotometer.

RESULTSIn KKAy mice, blood levels of FBG, TG, TC, ALT, AST and liver cell apoptosis rate were higher; the bax mRNA expression was higher and bcl-2 mRNA was lower markedly; the activities of SOD, GSH-Px, Na(+)-K(+)-ATPase in liver tissue were lower, and MDA content was higher than those in the normal control significantly (all P <0.01). Besides, mitochondria swelling and damage were found in liver tissue. While in the puerarin group after treatment, all the above-mentioned changes were alleviated to some extent.

CONCLUSIONSObvious liver injury emerges in KKAy mice. Puerarin shows a protective effect on the T2DM caused oxidative damage by way of up-regulating bcl-2 to inhibit oxidative stress, and improving the energy metabolic dysfunction in liver of mice.

Animals ; Diabetes Mellitus, Experimental ; complications ; drug therapy ; pathology ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; pathology ; Isoflavones ; therapeutic use ; Liver ; metabolism ; physiopathology ; Liver Diseases ; drug therapy ; etiology ; Male ; Mice ; Mice, Inbred C57BL ; Mitochondria, Liver ; pathology ; Oxidative Stress ; Phytotherapy ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2

Objective To evaluate the feasibility and safety of performing endoscopic retrograde cholangiopancreatography (ERCP) assisted by single-balloon enteroscopy (SBE) in patients with biliary obstruction after gastrointestinal reconstruction.Methods Clinical data about 7 cases of single-balloon enteroscopy assisted-ERCP (SBE-ERCP) were summarized including the completion of treatment,operation time,and complications for retrospective study.Results The papilla or anastomotic site was reached and therapeutic ERCP were performed successfully in 6 patients.The overall success rate was 85.7％ (6/7),and the mean operation time of SBE-ERCP was 42 min (ranging from 28 to 72 min).The afferent loop and papilla were failed to be confirmed in 1 patient.No complication such as perforation,pancreatitis or bleeding ocurred in all the 7 patients.Conclusion Single-balloon enteroscopy assisted-ERCP (SBE-ERCP) is feasible and relatively safe in postsurgical patients with gastrointestinal reconstruction.

Objective To analyze the distribution and drug resistance of pathogens isolated from blood culture specimens in a grade 3A hospital so as to provide a basis for the prevention and control of bloodstream infection ,and rational antibacterial drugs use .Methods A total of 3 241 blood culture results in our center from June 2014 to September 2016 were analyzed retrospectively . The blood samples were cultivated with the BacT/ALERT 3D and VersaTrek instruments .The bacterial identification and antibiotic susceptibility tests were conducted with VITEK‐2 Compact and ARIS 2X instruments .Results Among 3 241 blood culture speci‐mens ,99 strains of pathogenic bateria were isolated with the positive rate of 3 .1% ,including 42(42 .4% ) strains of Gram‐negative bacteria ,54(54 .5% ) strains of Gram‐positive bacteria and 3(3 .0% ) strains of fungus .The top three of Gram‐negative bacteria were in turn Eescherichia coli ,Klebsiella pneumonia and Pseudomonas aeruginosa ;the top three of Gram‐positive bacteria were in turn coagulase‐negative staphylococcus (CNS ) ,Staphylococcus aureus and streptococcus pyogenes .Enterobacteriaceae remained 100 .0% sensitivity to imipenem ,meropenem ,piperacillin/tazobactam and amikacin .The resistance rate of Gram‐positive bacterium to vancomycin ,linezolid ,teicoplanin and gentamycin was 0 .0% ,the rest had different degrees of drug resistance .Conclusion Ee‐scherichia coli ,CNS and Staphylococcus aureus are the most frequent pathogenic bacteria of the blood stream infection in our hospi‐tal .Pathogens show different resistance to different kinds of antibacterial agents .Clinic should rationally select the antibacterial a‐gents according to the drug susceptibility test results so as to slow down the generation of drug resistance bacterial strains .

Objective To investigate the relationship between mtND4 point mutation in sperms and asthenospermia. Methods Fifty-six asthenospermia cases and 44 control cases were collected using the WHO criterion for defining asthenospermia, the regions of mtND4 gene were amplified by using PCR of 3 pairs primers. Consequently, the point mutation, missense mutation and multiple single nucleotide polymorphisms (SNP) were analyzed by employing sequencing technology and bioinformatics tools. Results Six mutations never before identified were found. The frequency of single point mutation T10873C and T11944C in the control group were significantly higher than those in the asthenospermia group (P＜0.05). Eight cases involved T10873C or T11944C among the 10 cases in control groups with missense mutations were found. But, there were only 2 cases with such mutation in the 10 asthenospermia cases with missense mutations (P＜0.05). The previous 20 cases of missense mutations can be described as either multiple SNP group (with T10873C or T11944C) or nonmultiple SNP group. The percentage of a range and a plus b range of multiple SNP group of sperm was significantly higher than the non-multiple SNP group(P＜0.05). Conclusions mtND4 gene mutation, especially the missense mutation may induce loss of sperm motility. The mutations of T10873C and T11944C may be useful for sperm motility or counteract the influence for the sperm motility caused by these harmful mutations.

Objective To investigate sequence variations of 12 miRNA genes in multiple myeloma(MM) in order to find whether sequence variations in miRNA genes are associated with tumorigenesis and discuss the clinical significance of MM associated with miRNA genes mutations. Methods The miRNA gene mutations in 20 cases of MM, 4 MM-derived cell lines and 20 controls were detected by the methods of polymerase chain reaction single stranded conformation polymorphism (PCR-SSCP) and silver staining technique. Both clinical features and laboratory results were analyzed simultaneously. Results The electrophoretic patterns showed a total of three variations in miR-19a, miR-19b and miRNA-335,which were observed in 3 MM cells (15 %, 3/20). We also found variations of miRNA-335 in MM-derived cell lines KM-3and RPMI8226. However, no sequence alteration in the miRNA genes was observed in our set of controls. One of the three MM patients died, and two of them were detected mutations at the terminal stage of the disease.Conclusion A relative high frequency of miRNA gene mutation was found in MM and MM derived cell lines, which suggests possibility of a main mechanism underlying tumorigenesis. And, detecting miRNA gene mutations in MM might be benefit to evaluate the progression and prognosis of disease.

Objective To investigate the reversal effect of Topo Ⅱα-shRNA and Topo Ⅱβ-shRNA on Topo Ⅱ gene in K562/AS2 cells. Methods Three pieces of Topo Ⅱα-shRNA and three pieces of Topo Ⅱβ-shRNA were designed,synthesized and transfected into K562/AS2 cells by liposome. Expression level of Topo Ⅱα and Topo Ⅱβ mRNA were determined by real time fluorescence quantitative PCR. The expressions of Topo Ⅱα and Topo Ⅱβ protein were assayed with flow cytometer. Results After treated with Topo Ⅱα-shRNA or Topo Ⅱβ-shRN A for 24 hours,the expression level of Topo Ⅱα mRNA and Topo Ⅱβ mRNA protein in K562/AS2 cells decreased at most (78.22±0.01) %,(31.17±1.27) % (P <0.05),and (57.36±0.01)%,(23.98±1.22) % (P <0.05) respectively. Conclusion The expression of Topo Ⅱ gene can be down-regulated after infected with Topo Ⅱ-shRNA in K562/ AS2 cell line.