Objective: To study the mechanism of K562 leukemia cells proliferation and apoptosis induced by emodin. Methods: Inhibitory effect of emodin on proliferation of K562 leukemia cells was assayed by MTT method. The morphologic changes of K562 cells were observed under microscop after Wight-Giemsa staining. Apoptosis was checked by Annexin V/PI. The changes of cell cycle and apoptosis were detected by flow cytometry. The activities of Caspase-3,8,9 were checked by chromatometry to analyze the apoptosis of K562 cells. Results: The proliferation of K562 leukemia cells was significantly inhibited by emodin. The IC50 of K562 cells inhibited with emodin for 24 h,48 h and 72 h were 80,50 and 40 ?mol/L respectively. Typical morphological changes of K562 cells were observed by microscopy. The proliferation of K562 cells was obviously inhibited in dose-dependent manner. In Annexin V/PI,emodin induced K562 cells toward apoptosis in dose-dependent manner(P

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.

Acute promyelocytic leukemia (APL) is characterized by the generation of the prototypic promyelocytic leukemia-retinoicacid receptor alpha (PML-RARα), an oncogenic fusion protein due to chromosomal translocation. In a human myeloid cell line,PML-RARα is cleaved by neutrophil elastase (NE) to produce the mutational PML [nuclear localization signal (NLS) deletion andRARα (NLS-RARα, containing NLS of PML), both of which may play an important role in APL pathogenesis. The yeast two-hybridtechnique was used to screen the intracellular proteins interacting with NLS-RARα, which may be involved in NLS-RARα signaling. The NLS-RARα coding sequence was amplified by polymerase chain reaction method and was cloned into the bait plasmid pGBKT7vector, which, after the confirmation by sequencing, was transformed into yeast AH109 and the subsequent expression of bait plasmidwas proved by Western-blot. The transformed yeast AH109 was mated with yeast Y187 (containing leukemia cDNA library plasmidspACT2) in medium. Diploid yeast was plated on synthetic dropout nutrient medium containing X-α-gal for screening. After beingreintroduced into yeast AH109 and sequenced to verify the expression of ORF, eight positive colonies were obtained, among whichonecontaining JTV-1 was cloned. The interaction between NLS-RARα and JTV-1 was further supported by indirect immunofluorescence,GST pull-down and co-immunoprecipitation, respectively. These findings brought some new clues for the further exploration ofNLS-RARα signaling to APL.

Objective:To understand the biological characteristics, identification methods, genome structure and clinical significance of a rare strain of Aureimonas altamirensis isolated from clinical blood sample. Methods:The culture and biochemical characteristics of a strain isolated from blood culture were observed. The routine biochemical identification methods, MALDI-TOF mass spectrometer and 16S rRNA gene sequencing were used to identify the isolate. A phylogenetic tree based on the 16S rRNA gene sequences of the isolate and related strains was constructed. The genome of the isolated strain was sequenced and assembled, and gene prediction and functional annotation were made using related software. Phylogenetic analysis based on 31 house-keeping genes and genome-wide average nucleotide identity (ANI) analysis were conducted between the isolate and other Aureimonas sp. strains.Results:The isolated bacteria were gram-negative bacillus positive for catalase, urease and oxidase. It grew slowly on blood plate and could not be reliably identified by automatic bacterial biochemical identification systems or MALDI-TOF mass spectrometry. Results of the 16S rRNA gene sequencing and phylogenetic tree showed that the 16S rRNA gene sequence of the isolated strain was highly homologous to the strains of Aureimonas altamirensis NML070722 and IARI-ABL-26 (GenBank accession number: EU442518.1 and KC581669.1) in GenBank, and the gene sequence similarity was 99.93%. The total genome (National Microbiology Data Center genome accession number: NMDC60043566) length was 4 332 458 bp and GC content was 65.14%. There were 4 088 protein-coding genes and functional gene annotation showed that functional genes were mainly enriched in protein, amino acid, carbohydrate transport and metabolic functional regions. Pathogenic gene analysis predicted two high reliable virulence factor genes, but no drug resistance genes. House-keeping gene phylogenetic tree analysis showed that this strain was highly homologous to Aureimonas altamirensis strains of DSM21988 and C2P003 (GenBank accession number: GCF 001463885.1 and GCF 000800175.1), but ANI analysis showed that its genome was significantly different from those of the two strains. Conclusions:A rare strain of Aureimonas altamirensis was isolated from clinical specimen in China. As the biological and genomic characteristics of Aureimonas altamirensis had not been fully recognized, it was difficult to be correctly identified by conventional methods. The pathogenicity of Aureimonas altamirensis to immunocompromised patients and the significance of isolation in clinical specimens might need more case studies.

Objective:To construct a death education intervention program for advanced cancer caregivers to improve the reference for death education for advanced cancer caregivers.Methods:Content analysis, semi-structured interview, Delphi expert consultation method were used to develop a preliminary death education program based on the theory of knowledge, belief, and behavior. From April to May 2022, fifteen experts from palliative care, life and death education, oncology nursing, psychological nursing and other related fields were selected for two rounds of expert consultation, and the contents of the program were revised and improved through preliminary experiments.Results:After two rounds of expert consultation, the results showed that the expert opinions tend to be unanimous. The authoritative coefficient of experts was 0.87, and the Kendall coordination coefficients of feasibility, validity and scientificity of the two rounds of consultation were 0.181, 0.303, 0.363 and 0.249, 0.355, 0.366, respectively (both P<0.05). The preliminary experiments revised and improved the intervention frequency and content, and finally formed a death education intervention program for advanced cancer caregivers which included four-stage progressive death themes: made an appointment with death, made a discussion on death, made an embrace with death and made friends with death. Conclusions:The process of constructing a death education program for advanced cancer caregivers is scientific, and the content is feasible, valid, and scientific. In addition, it is of great significance to promote death education in palliative care.

Objective To access the prevalence of type 2 diabetes mellitus ( T2DM) and its associated risk factors among adults with obesity in Zhejiang province. Methods The enrolled subjects were selected among local residents aged≥18 years with body mass index≥28 kg/m2 from 15 counties by multi stage stratified cluster random sampling from July to November, 2010. Each participant was required to attend complete questionnaire, physical examination, and testing overnight fasting blood specimen. Results A total of 1 351 residents were enrolled, including 613 males and 738 females. The prevalence of T2DM in adult population with obesity was 15. 03%, being 14. 03% in male, and 15. 85% in female;and that in urban area was 16. 64%, and in rural area was 13. 93%. Data from multivariable logistic regression showed that factors such as ageing (OR=1. 473, 95% CI 1. 243-1. 747), a family history of T2DM(OR=8. 945, 95% CI 5. 481-14. 598), staple food intake (OR=1. 185, 95% CI 1. 017-1. 380), triglyceride(≥1. 7 mmol/L, OR=1. 542, 95%CI 1. 066-2. 232) were risk factors of T2DM;while annual income(OR=0. 695, 95%CI 0. 544-0. 888), and milk intake(OR=0. 750, 95%CI 0. 567-0. 993) were shown as protective factors. Conclusion The prevalence of T2DM in adults with obesity was raised, ageing, a family history of T2DM, staple food intake, and dyslipidemia appeared to be major risk factors for T2DM.

Objective:To study the genotyping characteristics of Brucella strains isolated from Lianyungang City (non-brucellosis epidemic area) of Jiangsu Province. Methods:Preliminary identification of 13 suspected strains of Brucella isolated from blood culture in Clinical Microbiology Laboratory of the First People's Hospital of Lianyungang City in 2018 was conducted; at the same time, the specific gene bcsp31 and insertion sequence IS-711 of Brucella were detected by quantitative real-time PCR (Real-time PCR), and the identification results were rechecked and typed. Multiple locus variable-number tandem repeat analysis (MLVA) was applied for genotyping, and the sequencing results were edited by Mega 4.0 software. Results:All the 13 strains were identified as Brucella by preliminary identification. Real-time PCR confirmed that all the 13 strains were Brucella melitensis. The results of MLVA showed that 13 strains of Brucella melitensis were divided into 12 genotypes and clustered in the "middle Mediterranean cluster". Among 13 strains of Brucella melitensis, 3 strains were biovar 1, 2 strains were biovar 2 and 8 strains were biovar 3. Conclusion:All the Brucella strains isolated from Lianyungang City are Brucella melitensis and the MLVA cluster is in the "middle Mediterranean cluster".

Objective:To predict and evaluate the antibacterial efficacy of linezolid, teicoplanin and daptomycin against Staphylococci bloodstream infections with Monte Carlo simulation, and to optimize the clinical administration program. Methods:A total of 1 847 Staphylococci strains isolated from blood samples between January 2018 to December 2019 were collected with the help of the Blood Bacterial Resistant Investigation Collaborative System (BRICS). Minimum inhibitory concentrations (MIC) of linezolid and daptomycin were detected by broth dilution method, while MIC of teicoplanin were detected by agar dilution method. The dosage regimens of linezolid were 800 mg once daily, 500 mg once every 12 hours, 600 mg once every 12 hours and 600 mg once every eight hours. The dosage regimens of teicoplanin were 400 mg once every 12 hours, 600 mg once every 12 hours, 800 mg once every 12 hours, and 1 000 mg once every 12 hours. The dosage regimens of daptomycin were 4 mg·kg -1·d -1, 6 mg·kg -1·d -1, 8 mg·kg -1·d -1, 10 mg·kg -1·d -1and 12 mg·kg -1·d -1. The probability of target attainment (PTA) and cumulative fraction of response (CFR) of three different dosage regimens were calculated by Monte Carlo simulation. A dosage regimen with CFR≥90.0% was a reasonable choice for empirical antimicrobial therapy. Results:PTA of linezolid against Staphylococci when MIC≤0.500 mg/L at four dosage regimens (800 mg once daily, 500 mg once every 12 hours, 600 mg once every 12 hours and 600 mg once every eight hours) were all over 90.0%. When MIC was 1.000 mg/L, the PTA of linezolid against Staphylococci under the dosages of 500 mg once every 12 hours, 600 mg once every 12 hours and 600 mg once every eight hours were 92.2%, 96.6% and 97.6%, respectively. The CFR of the four dosage regimens of linezolid were 73.9%, 83.7%, 90.8% and 95.3%, respectively. When MIC≤1.000 mg/L, PTA of teicoplanin against Staphylococci were all 100.0% at four dosage regimens (400 mg once every 12 hours, 600 mg once every 12 hours, 800 mg once every 12 hours and 1 000 mg once every 12 hours). When MIC was 2.000 mg/L, the PTA of teicoplanin (800 mg once every 12 hours and 1 000 mg once every 12 hours) against Staphylococci were both 100.0%. The CFR of the four dosage regimens of teicoplanin were 90.8%, 92.8%, 93.5% and 94.6%, respectively. When MIC≤0.500 mg/L, PTA of daptomycin against Staphylococci under the five dosages of 4 mg·kg -1·d -1, 6 mg·kg -1·d -1, 8 mg·kg -1·d -1, 10 mg·kg -1·d -1 and 12 mg·kg -1·d -1 were all over 90.0%. When MIC was 1.000 mg/L, the PTA of daptomycin against Staphylococci under the three dosages of 8 mg·kg -1·d -1, 10 mg·kg -1·d -1 and 12 mg·kg -1·d -1were 96.9%, 100.0% and 100.0%, respectively. The CFR of the five dosage regimens of daptomycin against Staphylococci were 97.4%, 99.2%, 99.9%, 100.0% and 100.0%, respectively. Conclusions:Linezolid (600 mg once every 12 hours), teicoplanin (400 mg once every 12 hours) and daptomycin (4 mg·kg -1·d -1) can achieve satisfactory antibacterial activity for Staphylococci bloodstream infections.

ObjectiveTo study the epidemiological characteristics of diabetes mellitus and awareness rate in Zhejiang adults.MethodA total of 17 437 adults from 15 counties were randomly selected with stratified multi-stage cluster sampling method from July to November,2010.Each participant was required to complete a set of standardized questionnaire,physical examination,and blood specimen collection.ResultsThe overall standardized rate of diabetes mellitus was 5.94％ ( crude prevalence 8.80％ ).It was increased by 96.67％ during past 8 years,as compared with the prevalence 3.02％ in 2002.The standardized rate of diabetes mellitus of the urban and rural area were 7.52％ and 5.19％ ( crude prevalence 11.33％ and 7.09％ ),respectively.The standardized rate of diabetes mellitus in the urban was higher than rural ( u =6.58,P＜0.05 ).The standardized rate of diabetes mellitus of the male and the female were 5.74％ and 6.15％ ( crude prevalence 8.36％ and 9.13％ ),respectively,without significant difference ( u =1.39,P＞0.05 ).The awareness rate of diabetes mellitus was 59.19％ ( 56.66％ in male and 61.23 ％ in female,x2 =3.26,P＞0.05 ).The awareness rate of the urban was higher than the rural (63.47％ vs 54.69％,x2=12.20,P＜0.01 ).ConclusionThe prevalence of diabetes mellitus showed a rapidly rising trend in Zhejiang province.The effective intervention should be taken at its early stage.

OBJECTIVEUsing multilevel analysis model to analyze the regional-cluster and risk factors of blood pressure among adult residents in Zhejiang province.

METHODSTotally, 17 437 residents were studied through a multi-stage cluster random sampling method from 60 streets/townships. Risk factors on SBP and DBP were analyzed through a two-level regression model, respectively.

RESULTSThe average blood pressure in males appeared as: SBP (125.14±17.90) mmHg, DBP (80.02±10.07) mmHg, compared with SBP (123.36±20.20) mmHg, DBP (77.42±10.09) mmHg in females. The average blood pressure in urban areas were SBP (126.29 ± 19.46) mmHg, DBP (79.64±10.20) mmHg, compared with SBP (122.82±18.86) mmHg, DBP (77.99±10.08) mmHg in rural areas, with differences statistically significant (P < 0.001), between the two areas. Pearson correlation coefficients between SBP, SDP and age were 0.44 and 0.21(P < 0.001), respectively. 5.24% variation of the SBP and 4.96% variation of the DBP were attributed to the environmental factors. Factors as age, gender, hypertension in family history, taking medication for blood pressure, physical exercise, smoking, alcohol drinking, BMI, waist, FPG, TG and TC at the individual level. were associated with both SBP and SDP (P < 0.05). Family income seemed to be related to SBP. Education, reducing salt intake, intensity of occupation would affect on DBP. Influence of FPG and TC on SBP varied among streets/townships while relation between TC and DBP also varied.

CONCLUSIONThe distribution of blood pressure showed regional-cluster phenomenon. Strategies for different regions directed to prevent hypertension should consider the individual factors such as age, BMI, FPG, TG, TC, the regional factors and the sensitivity of different population to factors.

Adolescent ; Bias ; Blood Pressure ; physiology ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Hypertension ; epidemiology ; prevention & control ; Male ; Middle Aged ; Multilevel Analysis ; Regression Analysis ; Risk Factors