A method was developed for purification of 20S proteasome (20S core particle, CP) by combining differential centrifugations with nondenaturing polyacrylamide gel electrophoresis (native-PAGE), irrespective of species origins of CPs. CP purified from human erythrocytes was subjected to proteomic analysis by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), revealing 33 spots of subunit isoforms with different molecular weights and isoelectric points, more than 14 constituent subunits. Furthermore, other four CPs were purified from yeast, mouse liver, two pancreatic cancer cell lines SW1990 and PANC-1 using this method mentioned above, and subjected to proteasome heterogeneity analysis by native/SDS-PAGE (native/sodium dodecyl sulphate polyacrylamide gel electrophoresis), together with CP from erythrocytes. The method described acts as a rapid and effective tool for CP isolations, and the results obtained may be served as a footstone for the investigations of species-dependent proteasome heterogeneity.

Objective To investigate the antibacterial activity of combined antimicrobial agents for imipenem-resistant Acinetobacter baumannii in vitro,so as to provide evidence for guiding clinical practice.Methods Twenty-seven isolates of irnipenem resistant Acinetobacter baumannii were collected from March,2007 to February.2008.The minimal inhibitory concentrations(MIC)of the isolates against eleven frequently used antibiotics were determined by E test and agar dilution.Combined drug sensitivity testing was performed by broth checkerboard titration assay.Synergism between minocycline and cefoperazone-sulbaetam was analyzed by time-killing curve.Results The results from checkerboard titration assay and time-kill studies showed that the synergy rates between minocycline and cefperazone-sulbactam,minocycline and amikacin,minocycline and ciprofloxacin were 74.1%,28.0%and 48.1%,respectively.The synergy rates between amikacin and cefoperazone-sulbactam,anaikacin and ciprofloxaein were 23.1%and 37.0%,respectively.The synergy rate between cefoperazone-sulbactam and ciprofloxacin was 7.1%.With combination therapy.both MICs of cefoperazone-sulbactam and MIC of minocyeline decreased to lower than breakpoints.According to time-killing curve,the synergy rate between cefoperazone-sulbactam and minocycline was 100%.Conclusion Minocycline and cefoperazone-sulbaetam can be used for treating infections caused by multidrug resistant Acinetobacter baumannii.

OBJECTIVE To get the message of drug sensitivity and characteristic of Candida infecting patients in our hospital. METHODS The CHROMagar candida color medium was applied to isolate and identify Candida strains.Drug sensitivity was analyzed by the K-B slip diffusion method. RESULTS Totally 393 strains of Candida in our hospital were isolated in the three years,including Candida albicans;C.tropicalis;C.glabrata;(C.krusei);Cryptococcus neoformans and the others;and their infection rate was 63.10%,12.47%,9.41%,8.40%,1.53%,and 5.09%,respectively.Among all strains,156(39.69%) strains were isolated from sputum,(66(16.79%))from stool,45(11.45%) from urine,38(9.67%)from leucorrhea,and 34(8.65%)from throat swabs.Sensitivity results to nine antifungals showed that 5-flucytosine,fluconazole,clotrimazole,amphotericin B,and nystatin had higher drug sensitivity to these fungi. CONCLUSIONS Isolated rate of Candida is higher from clinical samples.Drug sensitivity of different Candida to various antibiotics is different.Culturing and drug sensitivity of Candida should be put on important place and antibiotics should be used reasonably based on drug sensitive test to reduce drug resistance.

Mutagenesis of several male contraceptives in sperm bead anomalies was investigated. Results show that glycosides of tripterygium wilfordii hook (GTW) and its monomer T13, microwave induce sperm head anomalies. However, gossypol and monomer T4 and GTW do not induce sperm head anomalies. Adult male mice and rats were given orally GTW, monomer T4, T13 and gossypol. These chemical agents were delivered in 1% methylcellulose. Result indicated that frequency of abnormal sperm heads in GTW, T13 groups were significantly increased, while frequency of abnormal sperm heads in T4 and gossypol-treated animals were similar to that of normal controls. When male mice were exposed to microwaves of 0.5 kW for 1-2 min, for five weeks abnormal shape of spermatozoa could be found.

Objective To explore the correlation between the clinical nurses’turnover intention and their job satisfaction of their present working status.Method The Mueller/Mccloskey Satisfaction Scales(MMSS)and Turnover Intention Scale(TIS)were employed to implement the questionnaire survey among 531 registered nurses from a hospital in Taishan,Guangdong.Results The total scores on nurses turnover intention and their job satisfaction were(3?34±0?40)and(14?54±1?38)respectively.The opportunity for professional improvement,welfare and income,balance between job and family were negatively correlated with their turnover intention (all P<0?05).Conclusions The nursing administration should look into the conditions of nurse’s turnover intention and their job satisfactions so that they can take effective measures to improve their job satisfaction and decrease their turnover intention.

Objective To investigate the impact of 5-fluorouracil (5-FU) and cisplatin on miR-449b expression in human hepatocellular carcinoma (HCC) and elucidate the molecular mechanism of 5-FU and cisplatin inhibiting the migration of HCC cells.Methods Real-time qPCR analysis was conducted to determine the expression of miR-449b in 50 HCC tissues.RT-PCR assay was performed to detect the expression of miR-449b in HCC cells with 5-FU and cisplatin treatment.The migration of HCC cells with the overexpression of miR-449b was determined by wound-healing assay;Rescue assay was employed to investigate the correlation between 5-FU & cisplatin, miR-449b and the migration capacity of HCC cells;The putative targets of miR-449b were predicted and validated using target prediction programs and immunoblots.Results The expression of miR-449b decreased in HCC tissues (P<0.0001).miR-449b expression increased in HCC cells upon the treatment of 5-FU and cisplatin (P<0.001).The overexpression of miR-449b inhibited the migration of HCC cells (P<0.001).Rescue assay revealed that inhibition of miR-449b to prevent 5-FU and cisplatin induction resulted in suppressed migration in SMMC7721 cells(P<0.05).Catenin-δ was a functional target of miR-449b.Conclusions 5-FU and cisplatin inhibit the migration of HCC cells at least partly via inducing the expression of miR-449b.

AIM: To establish a model of subtotal nephrectomy (SNx) and investigate the changes of apoptosis and apoptosis-related genes (Bax, bcl-2, caspase-3, caspase-8 and caspase-9) in the rat remnant kidney. METHODS: Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at 1, 2, 4, 8, 12, 16, 26 and 40 weeks after operation. The tissues of remnant kidneys were collected to detect apoptosis cells by in situ end-labeling of cleaved DNA (TUNEL) and proliferating cells by determining the proliferating cell nuclear antigen (PCNA). The expression of Bax, bcl-2, caspase-3, caspase-8 and caspase-9 was measured by RT-PCR and Western blotting. The proteins were detected by immunohistochemistry staining. The relation between apoptosis, proliferation, glomerulosclerosis and renal interstitial fibrosis was also observed. RESULTS: The results showed the renal pathological dynamic changes in 5/6 nephrectomy remnant kidneys were tubule-interstitial inflammation and fibrosis, as well as glomerulosclerosis. There were transient increases in both proliferating and apoptotic processes in glomerulus, tubules and interstitium. Apoptosis was increased and most of apoptotic cells were detected in tubular epithelial cells and interstitial area. The mRNA and protein expression of Bax, caspase-3, caspase-8 and caspase-9 were increased in all course, and peaked at week 4 and 40 in the SNX rats. The successive changes of these parameters were parallel to the level of focal inflammation in interstitium. Glomerulosclerosis index was related with focal inflammation cells and 24 hours urine protein (r=0.788, 0.822; P

To look for new genes from human brain, get a fragment was obtained using adaptor primer and 3' anchor polymerase chain reaction (PCR) with the human adult whole brain cDNA as template. The fragment was cloned into T easy vector and automatically sequenced with 310 Genetic Analyzer. Later the whole length cDNA of this novel gene was got with the method of 3'rapid amplification of cDNA end (RACE). The whole length of cDNA of this novel gene is 2 024 bp. Chromsome location is at 14q11.2 including 16 extrons and 15 introns. After scanning the sequence against GenBank it is proved that the sequence is a new one. ORF analysis showed that there is a complete coding region in it,it can interprate a protein containing 357 amino acid residules. ProDom analysis result showed that there is an acyl carrier protein (ACP) like domain in it. The gene was banked into GenBank. Then, a pare of primers were designed and were used to amplify the coding region and cloned into pGEX-4T1 expressing vector to express it in E. coli . The Dot blotting and Northern blot showed that this novel gene is highly expressed in the normal adult human brain.

Objective To analyze the polymorphism of histidine rich protein 2(HRP II)gene in Plasmodium falciparum (Pfhrp2)from falciparum malaria patients in Yunnan Province,so as to lay the foundation for studying the defection of antigen genes of Plasmodium. Methods The filter paper blood samples and related information of falciparum malaria cases reported were obtained in Yunnan Province from August 2012 to September 2015. Under the guidance of the specific primers,the exon2 regions in Pfhrp2 gene in P. falciparum from DNA samples were amplified by PCR,and the PCR products were sequenced. The sequences of exon2 region in Pfhrp2 gene were blasted by comparing with the reference sequences AY816237,AY816240,and AY816301. Next,the polymorphism of the sequence in exon2 region of Pfhrp2 gene was analyzed by MEGA 5.04 software. The conserved sites and genetic distances between sequences were calculated by using the software as well,and the clustering tree was drawn according to the genetic distances between the amino acid sequences. Results A total of 218 bloods samples from the falciparum malaria cases in 15 prefectures of Yunnan Province were collected,and the sources of infection included Yun?nan,Africa and Myanmar. The PCR results showed that the exon2 regions in Pfhrp2 genes of 155 samples were positive by am?plification and their products were sequenced successfully. The sequence analysis showed that the length range of the amino acid residues of exon2 region in Pfhrp2 gene was from 115 aa to 298 aa,the average length was 239.7 aa. There was no statistically significance among the means of the amino acid residues of the isolates from Africa( 239.9 aa),Myanmar(239.5 aa)and Yun?nan(241.6 aa)(F=0.025,P>0.05). All the 155 amino acid sequences ended with type 12 repeat,98.1%(152/155)of them started with type 1 repeat and 1.9%(3/155)of them started with type 2. Type 2 presented most frequently repeat in all the se?quences and the average repeat times were 12.9. The homologous locus of the DNA sequences in exon2 regions of the 155 Pfhrp2 genes was 894 bp,among which the conservative sites accounted for 20.6%(186/894),and the variable sites for 78.2%(699/894). The genetic distances between the sequences of Africa isolates ranged from 0 to 0.741,and those of the Myanmar and Yun?nan isolates were 0-0.948 and 0-0.750,respectively. The cluster analysis showed that all the 155 sequences clustered into 3 cat?egories on genetic distances between amino acid sequences according to the size of the amino acid sequence length. At the same level,the sequences had approximate lengths and amino acid repeat types. Conclusion The sequence of exon2 region in Pfhrp2 gene of P. falciparum from falciparum malaria cases in Yunnan Province is highly polymorphic,the P. falciparum iso?lates are clustered mainly according to the size of the amino acid sequence of exon2 region in Pfhrp2 gene.

Objective To investigate the time distribution characteristics and the epidemic trends of imported malaria cases in Yunnan Province. Methods The malaria case records and epidemiological history data of Yunnan Province were collected, and the local infection cases were excluded. The data were statistical analyzed. Results The imported malaria cases had a sig-nificantly seasonal periodicity(Q=26.574,P<0.05)and epidemic trends(Q=35.487,P<0.05). The imported peak was in May,while February was the lowest month of imported cases,and the difference was significant(Z=-2.619,P<0.05). The simple seasonal prediction model was the best model(R2= 0.677,BIC = 4.867)for forecast while the residual sequence was white noise(Q=14.226,P>0.05). By using the model to predict the cases in January,February and March of 2016,the num-ber(95％CI)were 29(7-50),22(0-44)and 31(8-54),and the actual number of imported malaria cases were 29,24 and 38 cases respectively and all cases were included in the 95％CI. Conclusion The imported malaria cases in Yunnan Province had a significantly seasonal periodicity and epidemic trends,and the established model has good prediction on the recent cases.