Effect of trifluoro-icaritin on protein levels related to Akt/mTOR signaling pathway and autophagy in myocardial ischemia/reperfusion injury rats / 中国药理学与毒理学杂志

OBJECTIVE: To investigate the effect of trifluoro-icaritin (ICTF) on myocardial ischemia/reperfusion injury (MI/RI) in rats and to explore whether it plays a role in regulating autophagy through the serine/threonine kinase/mammalian target of rapamycin (Akt/mTOR) signaling pathway. METHODS: Male SD rats were ligated for 45 min and reperfused for 60 min to establish an MI/RI model. The rats were divided into sham, MI/RI model and model+ICTF 0.5, 1.0 and 2.0 mg·kg-1groups. II lead electrocardiogram (ECG) in T wave and ST segment changes were recorded. The area of myocardial infarction was determined by TTC. The protein levels and phosphorylation levels of microtubule-associated protein 2/1 light chain 3 (LC3 II/LC3 I), Beclin-1, Akt and mTOR in myocardial tissues were detected by Western blotting. The level of LC3 in myocardial tissues was detected by immunofluorescence test. RESULTS: The ECG showed that the T wave (P<0.05) and ST segment (P<0.01) of the model group were significantly higher than those of the sham group after 60 min of reperfusion, while the T wave (P<0.05) and ST segment (P<0.01) of the ICTF 1.0 mg·kg-1group were obviously lower than those of the model group. TTC staining of heart sections showed that the area of myocardial infarction in the model group was larger than in the sham group (P<0.01), while that in the ICTF 1.0 mg·kg1group was smaller than in the model group (P<0.01). Western blotting results showed that compared with the sham group, the ratios of LC3 II/LC3 I (P<0.01) and p-Beclin-1/Beclin-1 (P<0.05) in the model group were significantly increased, while Akt (P<0.01) and mTOR (P<0.05) were decreased. In addition, compared with the model group, the ratio of LC3 II/LC3 I (P<0.01) and p-Beclin-1/Beclin-1 (P<0.01) in the ICTF 1.0 mg·kg-1group was reduced, and the phosphorylation levels of Akt (P<0.01) and mTOR (P<0.01) were increased. Immunofluorescence results of frozen sections of myocardial tissues showed that LC3 protein expression increased in the model group compared with the sham group (P<0.01), but decreased in the ICTF 1.0 mg·kg-1group compared with the model group (P<0.01). CONCLUSION: ICTF has a protective effect on myocardial ischemia/reperfusion injury in rats, and its mechanism may be related to the regulation of Akt/mTOR signaling pathway to inhibit excessive autophagy.

Recent progress of positron radionuclide labeled small molecule inhibitors of prostate specific membrane antigen for PET/CT imaging of prostate cancer / 生物医学工程学杂志

Prostate cancer is the most common tumor of the urinary system, and its mortality rate is second only to lung cancer. With the specific and high expression on the surface of prostate cancer cells, prostate-specific membrane antigen (PSMA) has been an ideal theranostic target of prostate cancer with great clinical significance and research value. Positron emission tomography/computed tomography (PET/CT), a new modality of molecular imaging combining functional metabolic information and anatomical structure, provides high diagnostic performance for cancer detection. This paper mainly reviewed recent progress of PSMA inhibitors labeled by positron-emitting radionuclides for early diagnosis, preoperative staging, response assessment, restaging and metastasis detection of prostate cancer.

Correlation between pulsed-field gel electrophoresis profiles and Salmonella serotypes / 中华流行病学杂志

Objective To analyze the relationship between pulsed-field gel electrophoresis (PFGE) subtyping and serotyping of Salmonella (S.).Methods PFGE was performed and profiles were analyzed on 1230 Salmonella isolates which comprising the top five serotypes including Typhimurium,Enteritidis,Derby,Agona and Senftenberg identified in China.The potential predictive relationship between PFGE banding patterns and particular serotypes was compared and the discriminatory consensus band class markers of individual serotypes were identified.Results Among all the 1230 Salmonella strains,1149 strains were found assistant with serotyping through PFGE cluster analysis,providing the matching accuracy reaching 93.4％.For the five serotypes,the positive prediction rate appeared more than 90.0％ and the negative prediction rate was over 95.0％ on serotype cluster prediction.Conclusion Results presented in this study were representatives of the top 5 Salmonella serovars,showing that PFGE cluster analysis could provide clues to identity and confirmation of serotypes.

Molecular analysis on non-O1 and non-O139 Vibrio cholerae isolates / 中华流行病学杂志

Objective According to results from the two-month consecutive surveillance program in Maanshan,six suspected cases of non-O1 non-O139 Vibrio (V.) cholerae infection,were found that called for identification of pathogens as well as molecular-epidemiological analysis to determine the aggregation of the epidemic situation.Methods Biochemical and serotype identification,hemolysis test,and drug sensitive test were used to detect the drug resistance spectrum.Real-time PCR and conventional PCR were used to detect the presence of V.cholerae specific genes,virulent genes and its related genes,including ompW,ctx,tcpA,toxR,hlyA,zot,ace,rstR and g ⅢCTX.Pulsed-field gel electrophoresis (PFGE) was used to analyze the molecular type of strains.Results All the six isolates of non-O 1 non-O 139 V.cholerae were identified by biochemical and serologic tests,and appeared to be β hemolytic.Twelve out of the 14 kinds of drugs showed 100％ sensitive.All isolates were positive of ompW gene by real-time PCR,but negative for ctx,tcpA,zot,ace,rstR and gⅢ CTK.Five of the six isolates were positive for toxR and hlyA,except for strain 1001434446.All strains had different PFGE types,but two strains had similar types.All strains had a low similarity compared to the toxigenic V.cholerae.Conclusion Six cases ofnon-O1 and non-O139 nontoxigenic V.cholerae infection appeared in the same period.Along with epide(m)iological information,we noticed that these cases had a sporadic nature,but frequently appeared in the same area.We got the impression that public health measurements should be strengthened,with special attention paid to those diarrhea outbreaks caused by non-O 1 /non-O 139 strains since V.cholerae had appeared in low incidence.

Molecular subtyping of Vibrio cholerae isolates from outbreaks of cholera by pulsed-field gel electrophoresis in Hainan in 2008 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the molecular characteristics and genetic correlations of Vibrio cholerae isolates in Hainan in 2008, so as to provide pathogenic proof to diagnose the plague.</p><p><b>METHODS</b>Seventy six cholera strains were isolated from this cholera epidemic.69 strains were obtained from patients, 7 were isolated from external environment, among which, one was from patient's toilet, one from water sample, three were isolated from fish pond near patient's home, one came from swab of the patient vomit on the ground of health center and one from swab of kitchen knife from Hainan University canteen respectively. With conventional aetiological methods, pulse-field gel electrophoresis was conducted and the patterns of the 76 isolates were analyzed. The PFGE image was analyzed using BioNumerics (Version4.0, Applied Maths BVBA, Belium). Image bands were identified and similarity coefficient was automatically generated.</p><p><b>RESULTS</b>Seventy six strains were isolated from Vibrio cholerae outbreaks in Hainan in 2008.5 PFGE patterns of patient's isolates in June were the same, sharing a similarity coefficient of 100%. 70 PFGE patterns of patients and water in October and November were completely same, the similarity coefficient being 100%. But they were not same as that of June. 1 PFGE pattern of isolate from the sample in Hainan University was different, only sharing a similarity coefficient of 79.7%, which showed no correlation with the outbreak.</p><p><b>CONCLUSION</b>Different outbreaks of Vibrio cholera occurred in Hainan in 2008. The epidemic in October and November at different counties was one outbreak. The pollution of water in environment was an important factor for outbreak.</p>

Study on the epidemiological characteristics and molecular typing of Salmonella enterica subsp. enterica serovar Senftenberg in Shanghai / 中华流行病学杂志

ation were complicated, with the characteristics as the obvious decreasing number of patients, with no food-borne isolates in 2007.

Molecular characterization of Vibrio cholerae phage-type 6b epidemic isolates from 1998 to 2001 in Sichuan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the molecular characteristics of phage-type 6b isolates emerging in 1998-2001 cholera epidemics in Sichuan province.</p><p><b>METHODS</b>Isolates were analyzed by phage-typing, pulsed field gel electrophoresis (PFGE) and ompW gene sequencing.</p><p><b>RESULTS</b>All phage-type 1b and 6b isolates in Sichuan province from 1998 to 2001 were toxigenic. A total of 24 patterns were identified after PFGE analysis, and one predominant pattern consisted of 13 isolates. Several 1b and 6b isolates from Sichuan and isolates of the 1b from other provinces showed the same PFGE pattern. Mutation in ompW gene was found in 6b isolates.</p><p><b>CONCLUSION</b>V.cholerae O1 6b isolates in Sichuan province from 1998 to 2001 have special genetic markers, and might genetically correlate with contemporaneous 1b isolates.</p>

Study on the application of pulsed-field gel electrophoresis regarding infection sources identification during an outbreak of Vibrio cholerae in Jiangxi Province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the correlation between Vibrio cholerae strains isolated from natural enviroment and fishery products and the source of infection during V. cholerae outbreaks.</p><p><b>METHODS</b>Cholera toxin gene was detected by polymerase chain reaction (PCR) amplification. Pulsed-field gel electrophoresis (PFGE) was used to subtype the isolates. Results of PFGE were analyzed and clustered by BioNumerics software (Version 4.0).</p><p><b>RESULTS</b>During the outbreaks, a total number of thirty O139 V. cholerae related serogroups were collected from patients, carriers, sewage and fishery products were identified and proved to be toxigenic. They could be clustered into four PFGE patterns when digested by Not I. These two V. cholerae outbreaks were caused by the same source of infection because of the following reasons: (1) PFGE patterns of the predominant strains isolated from two outbreaks were identical; (2) they were identical to the PFGE patterns of the strains isolated from the green turtle and rana catesbiana which were bought from the same wholesale store.</p><p><b>CONCLUSION</b>Green turtle and rana catesbiana that were contaminated by toxigenic O139 V. cholerae strains seemed to be the source of infection causing the O139 V. cholerae outbreaks in Jiangxi province. Rapid laboratory surveillance and epidemiologic investigation were important in identifying the source of infection during the outbreaks of V. cholera.</p>

Establishment of fluorescent amplified fragment length polymorphism in Vibrio cholerae and evaluation in molecular typing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To develop fluorescent amplified fragment length polymorphism (AFLP) method and to evaluate the its typing capability with pulsed-field gel electrophoresis (PFGE) in molecular typing of Vibrio cholerae.</p><p><b>METHODS</b>Forty-seven strains of V. cholerae, with different PFGE patterns, were selected as the reference group to optimize the selective primers of AFLP analysis. Eighty-three strains including 20 strains from one epidemic episode, isolated from different provinces during 1961 and 2005, were used to compare the typing abilities of AFLP and PFGE. LI-COR4300 DNA sequencing system was used for AFLP electrophoresis. The images were recorded by Saga(MX) software and transferred to BioNumerics for clustering analysis. A standard protocol for V. cholerae from PulseNet was used in PFGE.</p><p><b>RESULTS</b>When comparison was made with different selective primers on AFLP based on the 47 strains, results showed that the optimized selective primer pair was EcoR I-G/Mse I-T, and the reproducibility of the tests was 99.2%. Eighty-three isolates showed 52 AFLP patterns and 44 PFGE patterns, with D values as 0.9545 (AFLP) and 0.9251 (PFGE) respectively.</p><p><b>CONCLUSION</b>The protocol of fluorescent AFLP on V. cholerae typing was established. AFLP was higher than PFGE in discrimination of V. cholerae which could be used for molecular typing. When combined with PFGE, AFLP became a more insightful tool to identify genome difference of different isolates.</p>

Molecular typing of Neisseria meningitidis serogroup C strains with pulsed field gel electrophoresis in China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the characteristics of epidemiology and molecular typing on Neisseria meningitidis serogroup C strains associated with outbreaks of Anhui province and sporadic cases in China, using pulsed field gel electrophoresis (PFGE).</p><p><b>METHODS</b>212 Neisseria meningitidis serogroup C strains were isolated from invasive meningococcal cases, close contacts and healthy carriers, including 48 strains from Anhui province with 38 strains associated with serogroup C outbreaks. PFGE were performed by genomic DNA digestion with Nhe I restriction enzyme. The results of PFGE were analyzed by BioNumerics software (Version 4.0, Applied Maths BVBA, Belgium).</p><p><b>RESULTS</b>A total number of 212 Neisseria meningitidis serogroup C isolates were typed by 43 patterns, named AH1 to AH43. In China, AH1 pattern was the major PFGE pattern with 69.3% (n = 147) of all strains, distributed in 11 provinces. Three types of PFGE patterns (AH1 to AH3) were found in 48 strains from Anhui province, in which, 93.8% (n = 45) belonged to AH1. 97.4% (n = 37) of 38 strains associated with serogroup C outbreaks in Anhui province showed AH1 pattern. A total of 53 serogroup C strains were isolated from invasive meningococcal cases with 67.9% (36/53) of AH pattern. 71.9% (87/121) of serogroup C strains isolated from contacts of invasive meningococcal cases was AH1 pattern and 63.2% (24/38) of the strains from healthy carriers showed AH1 pattern.</p><p><b>CONCLUSION</b>By PFGE typing and analysis, AH1 pattern of Neisseria meningitidis serogroup C strains was proved to be the main clone which causing the outbreaks in Anhui province and might be responsible for the sporadic serogroup C meningococcal disease epidemics else where in the country.</p>

Study of Immobilization of Yeast Alcohol Dehydrogenase Cross-linked Magnetic Chitosan Microspheres / 中国生物工程杂志

Magnetic chitosan microspheres(M-CS) were prepared and used for yeast alcohol dehydrogenase(YADH) immobilization.The optimum technology and the properties of immobilized YADH were studied.The optimal immobilization conditions for YADH were:20ml of 0.25mg/ml of YADH in phosphate buffer(0.05mol/L,pH 7.0) reacted with 50mg of magnetic M-CS at 4℃ for 2h.The microspheres were characterized by transmission electron microscopy,the results showed that M-CS were regular sphere with an average size of 30nm and had magnetic response characteristic.The M-CS suspended in H2O solution was easily precipitated and separated by magnetic field.Mechanical strength and crosslinking degree of M-CS were influenced by the ratio of carrier and immobilized YADH,ion concentration in phosphate buffer and pH of the solution.The immobilization was slightly influenced by the reaction temperature.The immobilization would improve its thermal,basic resistant and acid resistant stability.After the immobilized enzyme was kept between 35℃ to 75℃ for one hour,it still had 70 % of initial enzyme activity,when it was kept pH between 5.0 to 7.4 for one hour,it still had 80% of initial enzyme activity.The optimal reaction temperature of the immobilized enzyme was 40℃ compared to 30℃ of the free YADH,the optimal reaction pH of the immobilized enzyme was 6.8 as same as one of the free enzyme.Storaged at the temperature of 4℃ for 30 days without any protection by reagent,the free enzyme only kept 26% of the original activity but the immobilized enzyme still retained 60% of the activity.The immobilized enzyme maintained 70% of the activity after circular use 5 times.The Km value of immobilized YADH for Pyruvate was 2.58 mmol/L compared to 3.31 mmol/L of the free YADH,it would reduce its appetency for the substrate.

Immunoscreeening of Schistosoma japonicum schistosomula cDNA library with sera from Microtus fortis and bioinformatic analysis of novel genes / 中国血吸虫病防治杂志

Objective To obtain novel vaccine candidate antigens against Schistosoma japonicum. Methods S. japonicum schistosomula cDNA library was screened by using sera of Microtus fortis that was naturally resistant to schistosomiasis. The positive clones were transformated into Escherichia coli BM25.8, E. coli clones containing the plasmid cultured in LB, and then selected for plasmid extraction, the plasmid DNA was digested by EcoRⅠand Hind Ⅲ, and analysed by agarose gel electrophoresis. The positive clones were also sequenced and the data were analysed through the internet Nucleotide BLAST software of NCBI and Expert Protein Analysis system of GeneRunner and HNN. Results Twelve positive clones were obtained after repeatedly immunoscreening the library and their sizes ranged from 300 bp to 1100 bp. Two novel genes (named as Sj-sMf1 and Sj-sMf2) with complete ORF were obtained. The deduced protein of Sj-sMf1 consisted of 93 amino acids while Sj-sMf2 consisted of 61 amino acids. Sj-sMf1 protein predicted containing one cAMP phosphorylation site and Casein kinase C phosphorylation site, respectively. Sj-sMf1 protein predicted containing one Casein kinase C phosphorylation site and two Protein kinase C phosphorylation sites. Conclusion Two novel genes predictably encoding unknown proteins are obtained from immunoscreening of Schistosoma japonicum schistosomula cDNA library by M. fortis sera.