Brucellosis is a zoonotic disease of global importance for infecting humans .The early diagnosis of brucellosis infection plays a significant role in the treatment and rehabilitation .This paper reviews the different methods used to diagnose brucellosis ,particularly introduces the basic principles and applications of fluorescence polarisation assay as a diagnostic tool for brucellosis ,which could provide the reference for clinical diagnosis and epidemiological investigation on brucellosis .

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Objective To explore the effect and indication of splenectomy in liver transplantation.Methods From January 2001 to April 2006,260 patients underwent piggyback orthotopic liver transplantation(PBOLT),and 28 patients had undergone combined PBOLT and splenectomy(splenectomy group).These patients were compared to 56 randomly selected non-splenectomy patients from the same transplant period,meaningly two controls were se- lected for every non-spleneetomy case.Two groups were analyzed with respect to rate of infection and survival rate, as well as biopsy-proven acute allograft rejection within 30 days after transplantation.Results Rate of infection in the splenectomy group was higher than that in the non-splenectomy patients(85.7% vs 55.4%,P

BACKGROUND: Greatly importance has been attached to ceramic-on-ceramic bearing surface due to its excel ent wear resistance. But the risks of squeaking and ceramic fracture also go with it. Up til now, the choice between ceramic-on-ceramic and ceramic-on-polyethylene bearing surfaces in primary total hip arthroplasty remains controversial. OBJECTIVE: To compare the clinical outcomes and safety between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty based on meta analysis. METHODS: We electronical y searched databases including PubMed/Medline, Embase, Web of Science, Cochrane Col aboration database, Chinese Biomedical Literature Database (CBMdisc) and China National Knowledge Internet for randomized control ed trials on the comparison between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty from inception to January 2015. References of the included studies were also retrieved. Investigators severely selected the studies, extracted data and assessed the quality according to the inclusion and exclusion criteria. Then, meta-analysis was performed using RevMan 5.2 software. RESULTS AND CONCLUSION: Nine randomized control ed trials were included, involving 1 231 hips with ceramic-on-ceramic prosthesis and 932 hips with ceramic-on-polyethylene prosthesis. Meta analysis showed that both bearing surfaces achieved satisfied function recovery. But ceramic-on-ceramic had significantly increased risks of squeaking and ceramic fracture, meanwhile ceramic-on-polyethylene showed significantly higher wear rate. There were no significant differences in intra- or post-operative dislocation, osteolysis and other complications and prosthesis failure with any reason between two bearing surfaces. These results suggest that during the short- to mid-term fol ow-up period, no sufficient evidence can tel that ceramic-on-ceramic was obviously super than ceramic-on-polyethylene. Long-term fol ow-up is required for further evaluation.

Objective: To learn the awareness rate of adverse events following immunization ( AEFI ) among Chinese parents, so as to provide suggestions for promoting vaccination. Methods: We searched relevant articles published before 24th June, 2020 from CNKI, Wanfang, VIP, CBM, PubMed and Web of Science, calculated the pooled awareness rate and 95% confidence interval ( CI ) , conducted Egger＇s test for publication bias and sensitivity analysis for stability of results. Results: Eight articles using cross-sectional design were included after screening from 235 initial records. Among 5 433 subjects, the pooled awareness rate of AEFI was 66.76% ( 95%CI: 52.75%-78.33% ) . Non-immigrant population possessed a higher awareness rate ( 67.32% ) compared with the immigrant population ( 56.54% ) . The parents with different levels of education showed various awareness rate of AEFI ( P<0.05 ) . The awareness rates of "children should be observed for at least 30 minutes after vaccination","slight adverse effects were commonly seen after vaccination","local redness and induration might occur after diphtheria-tetanus-pertussis ( DTP ) immunization","polio vaccine might bring mild diarrhea" were 86.18%, 66.76%, 41.89% and 30.22%, respectively. Egger＇s test showed that there was no publication bias. Sensitivity analysis indicated that the results were robust. Conclusion The pooled awareness rate of AEFI among Chinese parents is 66.76%, with lower rates found in the parents who are immigrants and have lower level of education.

Hepatocellular carcinoma (HCC) is a common malignant tumor worldwise. The incidence rate of HCC is high and is easy to metastasis and recurrence, which seriously affects human health. Traditional chemical drugs have some challenges such as toxicity, side effects, and multidrug resistance, thus it is urgent to find new drugs and effective targets. Here we synthesized a novel chemical, protonic bis-phenanthroline (H-BP), and the antitumor effect was investigated in the study. The results showed that H-BP could selectively inhibit the proliferation of tumor cells and cause HCC apoptosis. And also, in HCC tumor-bearing mice, H-BP could effectively prevent the growth of tumor mass, even completely eliminate the tumor at medium dose (5 mg·kg^-1) and high dose (10 mg·kg^-1), and meanwhile H-BP has no significant effect on the body weight of mice. The experimental protocol was approved by the Animal Ethics Committee of Southwest University, and the experimental operation was strictly carried out in accordance with the ethical principles of animal use and care. Mechanism studies showed that H-BP induced HCC apoptosis was related to down-regulation the expression of pleomorphic adenoma gene like-2 (PLAGL2), a oncogene transcription factor, resulting in the down-regulation of PLAGL2 downstream proteins hypoxia inducible factor and β-catenin. This study not only introduces the dimerization method to form novel compounds that will provide a new approach for drug design, but also suggests that PLAGL2 may be an effective target in tumor therapy.

OBJECTIVETo investigate the apoptosis-promoting effect of PDCD5 on human prostate cancer cells PC-3M-1E8.

METHODSPCI-neo and PCI-neo-PDCD5 were transfected into PC-3M-1E8 cells by Lipofectamine 2000, the viability of the cells was analyzed by MTT assay 16 hours after removal of the serum, and the apoptosis was determined by in situ end-labeling and electron microscopy.

RESULTSThe viability and growing speed of the transfected cells were significantly decreased and their apoptotic indexes significantly increased as compared with the control group (P < 0.001).

CONCLUSIONPDCD5 may significantly inhibit the in vitro growth and promote the apoptosis of human prostate cancer cells PC-3M-1E8.

Apoptosis ; genetics ; physiology ; Apoptosis Regulatory Proteins ; genetics ; physiology ; Cell Line, Tumor ; Humans ; In Situ Nick-End Labeling ; Lipids ; chemistry ; Male ; Neoplasm Proteins ; genetics ; physiology ; Plasmids ; chemistry ; genetics ; Prostatic Neoplasms ; genetics ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods

OBJECTIVETo investigate the effects of electromagnetic radiation on the physiological indices and immune function of operators.

METHODSThe general conditions and electromagnetic radiation awareness rate of 205 operators under electromagnetic radiation were evaluated using a self-designed questionnaire. Physical examination, electrocardiography, and routine urine test were performed in these operators. Peripheral blood was collected from the operators under electromagnetic radiation for blood cell counting and biochemical testing, and their peripheral blood lymphocytes were cultured for determination of chromosomal aberrant frequency and micronucleus frequency. The data from these operators (exposure group) were compared with those of 95 ordinary individuals (control group).

RESULTSThe chief complaint of giddiness, tiredness, dizziness, and amnesia showed significant differences between the exposure group and control group (P < 0.01), and the difference in headache became larger with an increase in working years. The awareness rate of electromagnetic radiation damage was significantly higher in the exposure group than in the control group. The difference in bradycardia was significant between the two groups (P <0.01), and the incidence was higher with longer working years. Significant differences between the two groups were also found in the numbers of individuals with elevated alanine aminotransferase, total bilirubin, and direct bilirubin (P < 0.01), populations with increased lymphocyte ratio and decreased neutrophil ratio (P < 0.01), populations with positive occult blood, urobilinogen, and bilirubin tests, and the number of individuals with increased micronucleus frequency of cultured peripheral blood lymphocytes (P < 0.01). In addition, the exposure group had significantly increased complement C3 and C4 (P < 0.01), significantly increased IgG (P < 0.05), and significantly decreased IgM (P < 0.01), as compared with the control group.

CONCLUSIONElectromagnetic radiation may lead to the changes in physiological indices, genetic effects, and immune function and affect the health and immune function in operators. The adverse effects are increased as the working years increase. So it is important to strengthen occupational protection of operators under electromagnetic radiation.

Adult ; Chromosome Aberrations ; radiation effects ; Electromagnetic Radiation ; Female ; Humans ; Lymphocytes ; radiation effects ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Young Adult

OBJECTIVETo study the DNA methylation status of WT1 gene promoter region in hematologic malignancy cell lines and its correlation with WT1 gene expression.

METHODS1. RT-PCR and methylation-specific PCR were performed for detecting WT1 gene expression and DNA methylation status in its promoter region in 8226, HL-60, Jurkat, K562, KG-1, NB4, Raji, SHI-1, U266 and U937 cell lines. 2. Treatment of U937 cells with 5-aza-CdR, a demethylation inducing agent and the changes in WT1 gene expression level and its promoter region methylation status were determined.

RESULTS1. HL-60, K562, KG-1, NB4 and SHI-1 cells showed higher levels while 8226, Jurkat, Raji, U266 and U937 cells showed extremely low levels of WT1 expression. DNA hypermethylation in WT1 gene promoter region was identified in 8226, Jurkat, Raji, U266 and U937 cells. 2. The WT1 gene expression in U937 was enhanced after treatment with 5-aza-CdR accompanied with the decrease of methylated and the increase of unmethylated levels in its promoter region.

CONCLUSIONModulation of the DNA methylation status in WT1 promoter region is one of the epigenetic mechanisms for regulating its expression.

Cell Line, Tumor ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hematologic Neoplasms ; genetics ; Humans ; Promoter Regions, Genetic ; genetics ; WT1 Proteins ; genetics