BACKGROUND: Evidence on the relations of the American Heart Association's ideal cardiovascular health (ICH) with mortality in Asians is sparse, and the interaction between behavioral and medical metrics remained unclear. We aimed to fill the gaps. METHODS: A total of 198,164 participants without cancer and cardiovascular disease (CVD) were included from the China Kadoorie Biobank study (2004-2018), Dongfeng-Tongji cohort (2008-2018), and Kailuan study (2006-2019). Four behaviors (i.e., smoking, physical activity, diet, body mass index) and three medical factors (i.e., blood pressure, blood glucose, and blood lipid) were classified into poor, intermediate, and ideal levels (0, 1, and 2 points), which constituted 8-point behavioral, 6-point medical, and 14-point ICH scores. Results of Cox regression from three cohorts were pooled using random-effects models of meta-analysis. RESULTS: During about 2 million person-years, 20,176 deaths were recorded. After controlling for demographic characteristics and alcohol drinking, hazard ratios (95% confidence intervals) comparing ICH scores of 10-14 vs. 0-6 were 0.52 (0.41-0.67), 0.44 (0.37-0.53), 0.54 (0.45-0.66), and 0.86 (0.64-1.14) for all-cause, CVD, respiratory, and cancer mortality. A higher behavioral or medical score was independently associated with lower all-cause and CVD mortality among the total population and populations with different levels of behavioral or medical health equally, and no interaction was observed. CONCLUSIONS ICH was associated with lower all-cause, CVD, and respiratory mortality among Chinese adults. Both behavioral and medical health should be improved to prevent premature deaths.
Adult ; Humans ; Cardiovascular Diseases/prevention & control* ; East Asian People ; Prospective Studies ; Risk Factors ; Smoking

Objective To establish and preliminarily apply an effective PCR assay for detection of Tupaia(tree shrew)paramyxovirus(TPMV). Methods Using TPMV genomic DNA from NCBI GenBank, bases 8231 to 8720 were synthesized and inserted into a plasmid as a positive standard. One primer pair was designed based on this sequence. In total,60 respiratory swabs and 12 lung tissues from the tree shrews were tested in this PCR assay. Results A PCR method for detection of TPMV was successfully established,with high specificity and sensitivity of 11.5 × 10 -5μg/mL. PCR result testing 60 respiratory swabs and 12 lung tissues were negative. Conclusions PCR for detecting TPMV has good specificity and high sensitivity and can be used for conventional tree shrew paramyxovirus detection.

Objective: To study the specific killing effect of CD4 membrane protein targeted chimeric antigen receptor modified T (CAR-T) cell. Methods: The second generation CD4 targeted chimeric antigen receptor containing 4-1BB costimulation domain was insert into lentiviral vector through recombinant DNA technology. Lentivirus was prepared and packaged by 293T cells with four plasmids. Beads activated T cells were transduced with lentivirus and the transduction efficiency was checked with Protein L and flow cytometry. T cell subsets and IFN-γ concentrations were detected with probe-tagged antibody and cytometric bead assay. Results: ①The transduction efficiency of activated T cells with prepared lentivirus were 50.0%-70.0%. A subset of CD8⁺ T cell acquired dim expression of CD4 membrane protein after activation. CD4⁺T cell and CD8⁺CD4^dim T cell were gradually killed by CD4 targeted CAR-T post lentivirus transduction. ②The kill efficacy of CD4 targeted CAR-T cell and control T cell toward KARPAS 299 T cell at an E∶T ratio of 8∶1 for 24 h was (96.9±2.1)% and (11.2±3.1)%, CAR-T cell has a higher killing efficacy than control T cell (t=7.137, P=0.028). The IFN-γ concentrations in culture supernatant of CAR-T cell with K562-CD4 cell, CAR-T cell with K562 cell and CAR-T cell alone were (15 648±2 168), (1 978±354) and (1 785±268) pg/ml, CAR-T cell cocultured with K562-CD4 cell produced more IFN-γ than the other two controls (P<0.01). Conclusions CD4 targeted CAR-T has an immunophenotype of CD8⁺CD4^-T cell. CD4 targeted CAR-T cell has killing efficacy toward normal CD4⁺T cell and CD4⁺T lymphoma cell. CD4 targeted CAR-T cell also has a killing efficacy toward CD4^dim target cell.

Objective: To explore the impact of ticagrelor on platelet aggregation in patients with acute coronary syndrome (ACS) after percutaneous coronary intervention(PCI). Methods: A total of 98 ACS patients received PCI in our hospital from 2015-01 to 2015-12 were enrolled. The patients were randomly divided into 2 groups: Clopidogrel group, the patients received oral clopidogrel 300mg at first time and then maintained by 75mg/qd, n=48 and Ticagrelor group, the patients received oral ticagrelor 180mg at first time and then maintained by 90mg/bid, n=50. All patients were treated for 12 months.The level of vasodilator stimulated phosphoprotein (VASP) phosphorylation and platelet reactivity index (PRI) at pre-medication and 24h, 7 days and 1 month after PCI were detected; major adverse cardiovascular events (MACE) and bleeding events were recorded within 1 month after PCI, the incidence of platelet aggregation, MACE and bleeding events were compared between 2 groups.Results: The baseline information and PCI condition were similar between 2 groups, P>0.05. The overall average PRI was different between 2 groups, P<0.001 and PRI at each time point was different between 2 groups, P<0.001, different group and time point had interactive effect on PRI, P<0.001. Compared with Clopidogrel group, Ticagrelor group had the lower ratio of PRI≥50% at different time points after PCI, P<0.001. The incidence of MACE and bleeding event were similar between 2 groups within 1 month after PCI, P>0.05. Conclusion: Ticagrelor was superior toclopidogrel for anti-platelet aggregation in ACS patients after PCI, it didn't increase bleeding events.

Objective We established a rapid detection method of Pasteurella spp.and provided a reference for microbiological quality control of laboratory animal .Methods According to the β subunit of bacterial RNA polymerase ( rpoB) protein multiple alignments of 13 different Pasteurella spp.published in NCBI .The degenerate primers were designed by CODEHOP designer online .CODEHOP PCR method was applied to detecting Pasteurella spp.after the specificity and sensitivity of the method had been evaluated by 21 reference strains .Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5.The primers are able to distinguish between Pasteurella spp.and the other pathognic organisms of laboratory animal respiratory tracts .Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella.The Pasteurella positive rate was 19.1% in 609 animal ' s respiratory samples .The accuracy of positive results was 100%through verifying by sequenced and blast .Conclusions The established method has good specificity and sensitivity .It can be used to detect Pasteurella spp.in animal samples .

Objective To establish a detection technique for H.pylori(HP) infection in Mongolian gerbils using nested PCR technique.Methods H.pylori was cultured in vitro and inoculated into Mongolian gerbils.At the 10th week after infection, the HP in the gastric juice of Mongolian gerbil was detected by conventional PCR assay and the gastric juice, gastric mucosa, duodenal contents and colon stool were examined by nested PCR.Rapid urease test and ELISA were used to analyze the accuracy of the nested PCR assay.All of the PCR products were verified by sequencing.Results The positive rate of gastric juice detected by conventional PCR was 30%, while the positive rates of gastric juice, gastric mucosa, duodenal contents and colon stool detected by nested PCR were 100%, 100%, 90%, and 10%, respectively.The positive detection rates of rapid urease test and serum ELISA were 100% and 0%, respectively.Comparing the results of different methods, both the positive rates of gastric juice and gastric mucosa detected by nested PCR and the detection rate of rapid urease test were 100%, but the results of conventional PCR detection of gastric juice, the nested PCR detection result of stool in colon and of serum ELISA assay were lower than other methods.Conclusions Due to its high accuracy and sensitivity, the nested PCR assay of gastric juice can be used for the long-time detection of H.pylori infection in Mongolian gerbils, especially useful in the experiments of prevention and treatment of H.pylori infection.

Objective To understand the characteristics of minipigs infected withJapanese encephalitis virus(JEV).Methods After the brain tissues were treated, the pig brain tissue treatment solution was inoculated with BHK21 cells.Then, virus culture,indirect immunofluorescence assay, neutralization test, electron microscopic observation, and reverse transcription-polymerase chain reaction (RT-PCR) amplification of the new isolate E segment and PrM segment nucleotide sequence were performed and the genotype was identified.Results BHK21 cells were inoculated into 25 pigbrain tissues.Among them, three tissue-treated fluid couldinduce shrinkage and aggregation of BHK21 cells, and immunofluorescence staining showed strong green fluorescence response.The results of neutralization test showed that the neutralization titer of these three new isolates was 1:64, and the size of the virus particles was about 40nm under the electron microscope.The homology of both RT-PCR product sequencing results and E-segment of vaccine strain were 95%.Three new isolates were type GIII JEV.Conclusion The results ofthisstudydemonstrate that there is G III type Japanese encephalitis virus infection in the minipig farm.

Objective To acquire the prevalence and molecular identification data on Syphacia muris and provide reference for the revision of national standard. Methods 923 batches of 5199 SPF animals ( including one batch of 5 monkeys, 3 batches of 25 mini?pigs, 28 batches of 55 rabbits, 13 batches of 248 hamsters, 37 batches of 198 guinea pigs, 93 batches of 459 rats, 742 batches of 4179 mice, 5 batches of 25 chickens and one batch of 5 ducks) and 145 batches of 1389 clean animals ( including one batch of 3 rabbits, 4 batches of 31 hamsters, 16 batches of 157 guinea pigs, 32 batches of 268 rats and 92 batches of 930 mice ) came from 50 different manufactures in China. Direct microscopy real?time dynamic video recording techniques in combination with morphological identification method were applied to screen the Syphacia muris infestation. A multiple polymerase chain reaction ( multiple?PCR ) testing of the isolate based on amplification of the conserved portions of the Syphacia muris internal transcribed spacer (ITS), 28S ribosomal RNA (28S rRNA), NADH dehydrogenase subunits 1 (nad1) and cytochrome c oxidase subunit 1 (cox1) genes, and the molecular sequencing of the multiple?PCR amplicons was used to confirm the Syphacia muris infection. Results Syphacia muris eggs, larvae and adults were detected by using direct microscopy real?time dynamic video recording technique. Syphacia muris were detected based on the morphology and size of ovum, larvae, and female and male adult worms. Multiple?PCR and sequencing were performed to identify ITS, 28S rRNA, nad1 and cox1 genes of DNA extracted from the single egg, larva and adult parasite Syphacia muris. This approach allowed the specific identification with no amplicon being amplified from heterogeneous DNA samples, and sequencing confirmed the identity of the amplified sequences. Molecular characterization by multiple?PCR amplification and sequencing of the ITS, 28S rRNA, nad1 and cox1 genes demonstrated the presence of Syphacia muris. Multiple?PCR followed by sequencing confirmed 285 of 5199 SPF and 135 of 1389 clean animal samples classified as positive by using direct microscopy real?time dynamic video recording technique in the study as containing Syphacia muris?specific DNA. Comparison of the partial sequences of the ITS, 28S rRNA, nad1 and cox1 genes revealed 100% similarity amongst Syphacia muris from different animals. The prevalence of Syphacia muris infection in SPF and clean animals were 5?5% (285/5199) and 9?7% (135/1389), respectively. Conclusions Direct microscopy real?time dynamic video recording technique, multiple?PCR and sequencing can be used to rapidly detect and accurately identify Syphacia muris. The zoonotic nature of Syphacia muris can be regard as a public health alter, hence the good quality control of animal has an important role in protecting human health and safeguarding people safety. This is the first molecular identification and infection investigation of Syphacia muris in SPF and clean animals in China.

Objective To understand the Salmonella detectability in the laboratory animal testing laboratories, im-prove the level of detection for the quality of laboratory animals, by means of laboratory animals Salmonella proficiency tes-ting program.Method According to the proficiency testing program approved by CNAS, freeze-dried animal stool samples containing Salmonella bacteria and interference bacteria were prepared, and through stability and homogeneity tests quali-fied as proficiency testing samples.The randomly numbered samples were issued to the participating units by cold-chain transportation, and attached work instructions.The original reports and copies of the tests should be submitted on time.The sample results consistent with the results of the pretested results were considered as satisfactory, and the results inconsistent or fails to submit were judged as unsatisfactory.Results A total of 30 laboratories from 20 provinces and cities nationwide participated in this proficiency testing programs for Salmonella, including 28 ( 93.3%) laboratories with satisfactory re-sults, and two laboratories unsatisfactory ( 6.7%) .29 laboratories used separate culture methods, and two laboratories used PCR method.Conclusions The laboratory animal quality inspection agencies have good detection ability for Salmo-nella.The implementation of the capacity verification plan can well reflect the detection level of laboratories.

Objective To investigate the detection capacity of esterase-3 ( Es-3) in the laboratory animals monito-ring laboratories in China, and to improve the quality management of laboratories.Methods We prepared the test sam-ples according to the criteria of China National Accreditation Service for Conformity Assessment(CNAS), all the samples were certificated by homogeneity test and stability test.Then, samples with random numbers and standard operation instruc-tion were distributed to the participant laboratories.The laboratories should submit their reports before the deadline expires. When the results are the same as the standard results, the laboratories will receive excellent remark; when the results are the same as the standard results except the hybridization type, the laboratories will receive satisfactory remark;otherwise, it will receive unsatisfactory remark.If a laboratory did not submit report, the laboratory will also receive unsatisfactory re-mark.Results Ten laboratories participated in the program, and no laboratory received excellent remark.Nine laboratories (90.0%of enrolled laboratories) had satisfactory results, while one laboratory (10.0%of enrolled laboratories) had un-satisfactory results.Conclusions The nationwide overall detection level of laboratories in Es-3 is relatively high.Howev-er, some details should be noticed and several laboratories should improve their detecting ability.