ABSTRACT
A molecular diagnostic assay which could be stored at room temperature was developed to rapidly detect Mycobacterium tuberculosis (MTB) based on loop-mediated isothermal amplification (LAMP) technology and dry-reagent process. LAMP uses 4 or 6 primers and Bst DNA polymerase to amplify DNA at a constant temperature. The results showed that the LAMP assay could detect the amplification of IS6110 target gene within 20 min using real-time fluorescence signal detection. The sensitive of LAMP assay was similar to the PCR technology while the precision of PCR was better than LAMP (coefficient of variation, LAMP 18.9%, PCR 3.4%), meaning LAMP was more suitable for qualitative detection. The LAMP assay did not amplify DNA of other 10 types of pathogens, including Neisseria meningitidis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Rubivirus, mumps virus, adenovirus (type 3), adenovirus (type 7), respiratory syncytial virus B and parainfluenza virus type 2, indicating a good specificity. Furthermore, a dry-reagent assay was developed using air-drying and freeze-drying process. The performance of dried reagents did not change after 10 days storage at 50 ℃, meaning the dried reagents could be stored at room temperature (25 ℃) for more than six months. The dry-reagent LAMP assay also successfully amplified MTB DNA from several clinical samples within 20 min. In conclusion, the developed LAMP assay together with isothermal amplifier could rapidly detection MTB.
Subject(s)
Humans , Mycobacterium tuberculosis/genetics , Indicators and Reagents , Sensitivity and Specificity , Nucleic Acid Amplification Techniques/methods , DNAABSTRACT
A molecular diagnostic assay which could be stored at room temperature was developed to rapidly detect Mycobacterium tuberculosis (MTB) based on loop-mediated isothermal amplification (LAMP) technology and dry-reagent process. LAMP uses 4 or 6 primers and Bst DNA polymerase to amplify DNA at a constant temperature. The results showed that the LAMP assay could detect the amplification of IS6110 target gene within 20 min using real-time fluorescence signal detection. The sensitive of LAMP assay was similar to the PCR technology while the precision of PCR was better than LAMP (coefficient of variation, LAMP 18.9%, PCR 3.4%), meaning LAMP was more suitable for qualitative detection. The LAMP assay did not amplify DNA of other 10 types of pathogens, including Neisseria meningitidis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Rubivirus, mumps virus, adenovirus (type 3), adenovirus (type 7), respiratory syncytial virus B and parainfluenza virus type 2, indicating a good specificity. Furthermore, a dry-reagent assay was developed using air-drying and freeze-drying process. The performance of dried reagents did not change after 10 days storage at 50 ℃, meaning the dried reagents could be stored at room temperature (25 ℃) for more than six months. The dry-reagent LAMP assay also successfully amplified MTB DNA from several clinical samples within 20 min. In conclusion, the developed LAMP assay together with isothermal amplifier could rapidly detection MTB.
Subject(s)
Humans , Mycobacterium tuberculosis/genetics , Indicators and Reagents , Sensitivity and Specificity , Nucleic Acid Amplification Techniques/methods , DNAABSTRACT
@#AIM: To determine the relationship between interleukin-8(IL-8)levels in aqueous ocular samples and diabetic retinopathy(DR)through systematic evaluation and Meta-analysis. <p>METHODS: The PubMed, Embase and Web of Science database were searched from January 2010 to June 2021. A random effects model was used to combine the results, and the sensitivity analysis was performed to determine the stability and reliability of the arithmetic results, and subgroup analysis was used to identify possible sources of heterogeneity. <p>RESULTS: A total of 25 case-control studies were included. IL-8 levels in patients with DR were significantly higher than those in patients without DR(<i>SMD</i>: 1.57, 95%<i>CI</i>: 1.19-1.95, <i>P</i><0.01). Sensitivity analysis shows that the calculation results of random effects are stable and reliable. Subgroup analysis based on test method, region, sample source, and type of DR showed that the choice of these factors greatly influenced the relationship between IL-8 levels and patients with DR. Among them, the samples from Bead-based multivariate analysis(<i>I</i> 2=18%, <i>P</i>=0.27), Europe(<i>I</i> 2=38%, <i>P</i>=0.17)and nonproliferative diabetic retinopathy(NPDR)(<i>I</i> 2=0%, <i>P</i>=0.49)showed good consistency. ELISA, American, Asian, vitreous fluid, proliferative diabetic retinopathy(PDR)and other factors may increase the effect size.<p>CONCLUSION: Elevated IL-8 levels in aqueous eye solution are associated with the risk of DR, and IL-8 may serve as a potential predictor or therapeutic target for DR.
ABSTRACT
Diabetic retinopathy(DR), one of the common complications of diabetes, is a major cause of blindness. Traditionally, DR has been considered primarily a microvascular disease, and as research has progressed, it is now believed that disruption of the neuro-glia-vascular unit(NVU)and imbalance in its coupling mechanisms(coupling)play a key role in the early onset of DR. Understanding the cellular and molecular basis of NVU and how diabetes alters normal cellular communication and disrupts the cellular environment is important for the early prevention and treatment of DR. This paper summarizes the retinal NVU and its involvement in the molecular mechanism of DR pathogenesis, DR treatment based on retinal NVU repair, and discusses the future prospects and problems of DR.
ABSTRACT
@#Toll-like receptors(TLRs)are bridges of innate and acquired immunity. With the deepening of the research on the pathogenesis of dry eye, the induction and regulation of TLRs and signaling pathways in the immune response to dry eye have attracted more and more attention. Current studies have shown that the regulation of TLRs and signaling pathways helps to reduce or prevent the inflammatory response process in dry eye, so the regulation and control of TLR signaling pathways in the treatment and prevention of dry eyes has important significance. In this paper, TLRs and their signaling pathways are summarized in the pathogenesis of dry eye, and the effects of different intervention methods on the TLR pathway of dry eye are discussed, and the future development prospects and problems are also discussed.
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Objective To explore the consistency between identification diagnosis and pre-identification clinical diagnosis of patients with mental disorder undergoing forensic psychiatry identification. Methods The identification data of 1 369 appraised individuals who underwent criminal responsibility identification carried out by the Forensic Institute of Second Affiliated Hospital of Jining Medical University from 2014 to 2017 were collected retrospectively using self-designed investigation data sorting table. A comparative analysis of the mental disorder diagnosis results of expert opinion and past clinical diagnosis results was made. Results Among 1 369 appraised individuals, 964 cases （70.4%） were identified and diagnosed with mental disorder and 405 cases （29.6%） without mental disorder. Among the former, 63.3% （610 cases） were clinically diagnosed, which was higher than 43.2% （175 cases, P<0.05） in the latter. Among the various mental disorders that had been identified and diagnosed, patients with hysteria, stress, and neurosis had the highest proportion of clinical diagnoses （86.7%）, while patients with mental retardation had the lowest proportion of clinical diagnoses （9.6%）. Schizophrenia had the highest overall consistency rate of identification diagnosis and clinical diagnosis （98.4%）, while personality and behavior disorder had the lowest （33.3%）. The overall consistency rate between clinical diagnosis and identification diagnosis of the mental disorder group was 84.1%, and the Kappa value was 0.759. Compared with clinical diagnosis, the consistency rate between inpatient diagnosis and identification diagnosis was higher （85.9%, P<0.05）. Conclusion In forensic psychiatry identification that provides data of past clinical diagnosis and treatment, a high consistency between identification diagnosis and clinical diagnosis of the appraised individual who is identified and diagnosed with mental disorder exists. Clinical diagnosis （especially the inpatient diagnosis） has a relatively good reference value for forensic psychiatry identification.
Subject(s)
Humans , Criminals , Expert Testimony , Forensic Psychiatry , Mental Disorders/diagnosis , Psychotic Disorders , Reproducibility of Results , Retrospective StudiesABSTRACT
@#AIM: To investigate the relationship between single nucleotide polymorphism(SNP)in the TERT gene and risk of age-related macular degeneration(ARMD).<p>METHODS: Seven reported SNPs in TERT were selected and genotyped using SEQUENOM MassArray technology in 191 ARMD patients and 197 healthy normal controls. The correlations of the alleles, genotypes and the susceptibility of ARMD were evaluated by Logistic regression analysis.<p>RESULTS: The frequencies of SNP rs10069690 CC, CT, TT genotypes in ARMD patients were 64.4%, 31.4%, 4.2% compared with 74.6%, 22.8%, 2.5% in normal controls, respectively. Genetic model analysis revealed that SNP rs10069690 CT+TT genotypes were significantly associated with an increased risk of ARMD patients(<i>OR</i>=1.63, 95% <i>CI</i>=1.05-2.53).<p>CONCLUSION: SNP rs10069690 in the TERT gene is associated with the risk of ARMD.
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Objective To investigate the expression of integrin-linked kinase (ILK) and Toll-like receptor (TLR-2) in ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma and the effects on patients' outcomes.Methods Tissue specimens from 46 patients with MALT lymphoma were collected in this study.The expression of ILK and TLR-2 protein was detected by immunohistochemical methods,and the correlation of ILK and TLR-2 protein expression with clinicopathological features and patients' outcomes was analyzed.Results The positive rates of ILK and TLR-2 protein expression in tumor tissues were 67.4％ (31/46) and 71.7％ (33/46),respectively,which was related to the clinical stage of AnnArbor (P ＜ 0.05),rather than to sex,age and lesion location (all P ＞0.05).The survival of patients with ILK positive expression was less than that of ones with ILK negative expression [(21.5 ± 2.7)months vs.(29.2 ± 2.1) months] (P ＜ 0.05);meanwhile,the survival of patients with TLR-2 positive expression was less than that of ones with TLR-2 negative expression [(20.4 ±1.7) months v.(27.6 ± 2.3) months] (P ＜ 0.05).Conclusion ILK and TLR-2 are closely related to biological behavior of ocular adnexal MALT lymphoma,and combination detection of ILK and TLR-2 has a certain guiding value for diagnosis and prognosis.
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<p><b>OBJECTIVE</b>To study on the effects of sub-chronic exposures to high-frequency electromagnetic field (HF-EMF) on the estrous cycle, ovarian pathological changes and related hormones and preliminarily investigate the female genital toxicities of HF-EMF in rats.</p><p><b>METHODS</b>60 Wistar female adult rats were randomly divided into five groups based on body weight and radiated with different levels of 30 MHz electromagnetic field (0, 25, 100, 400 and 1600 V/m) for eight hours daily. Weekly the rats were continuously exposed five days. From the 48th day the four stage of estrus cycle were observed with smear method of the vaginal cell. Fifty-six days later the serum levels of sexual hormones were detected with the radioimmunoassay on estrus stage. The constituent ratio of the distinct follicle number on ovaries were observed with the HE staining and the ultrastructure was observed with the transmission electron microscope. Meanwhile, the ovarian humid weight and organ coefficient were observed.</p><p><b>RESULTS</b>There was no significant difference in ovarian humid weight and organ coefficient between the exposure and the control groups. The time of proestrus in the 100 V/m group, the 400 V/m group and the 1600 V/m group was decreased significantly [(15.00 +/- 5.06), (11.40 +/- 2.05) and (10.56 +/- 0.96) h in the exposure group compared with (18.70 +/- 2.96) h in the control group, P < 0.01], and the time of Anestrum in the 400 V/m group and the 1600 V/m group were increased significantly [(101.20 +/- 17.81) and (115.33 +/- 19.28) h in the exposure group compared with (69.80 +/- 11.42) h in the control group, P < 0.01)]. Serum LH in the 400 V/m and 1600 V/m group was increased significantly [(11.02 +/- 1.11) and (14.70 +/- 1.94) mU/ml in the exposure groups compared with (8.70 +/- 0.53) mU/ml in the control group, P < 0.01], and serum E2 was decreased significantly [(57.16 +/- 31.56) and (50.57 +/- 25.16) pg/ml in the exposure groups compared with (95.04 +/- 32.62) pg/ml in the control group, P < 0.01]. The composition ratio of the corpus luteum/albicans number in the 400 V/m group and the 1600 V/m group was increased significantly (19.75% and 19.04% in the exposure groups compared with 14.01% in the control group, P < 0.05). The composition ratio of the atretic follicle number was increased significantly in the 100 V/m, the 400 V/m and the 1600 V/m group (8.45%, 9.95% and 11.70% in the exposure groups compared with 7.72% in the control group, P < 0.01). The composition ratio of the mature follicle and the pri/sec follicle was decreased significantly in the 400 V/m and the 1600 V/m group (1.50% and 1.55% in the exposure groups compared with 3.36% in the control group. 22.24% and 21.09% in the exposure groups compared with 26.60% in the control group, P < 0.01). Along with the increase of radiation dose, the ultrastructure of cell on the ovaries appeared more abnormal.</p><p><b>CONCLUSIONS</b>The toxicities of female gonads are closely associated with exposures to HF-EMF. The nonage damage of female gonadal toxicities might emerge on the ovaries.</p>
Subject(s)
Animals , Female , Rats , Electromagnetic Fields , Estrous Cycle , Radiation Effects , Ovary , Pathology , Radiation Effects , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To analyze the toxicity of high-frequency electromagnetic field (HF-EMF) on the reproductive and endocrine functions of female workers.</p><p><b>METHODS</b>180 female workers exposed to HF-EMF for more than one year were selected as the exposure group, and 349 female workers in the marketplace were selected as the control group. Their menstrual status and pregnancy outcomes were evaluated with historical cohort study. Meanwhile, the serum levels of reproductive hormones including follicle stimulation hormone (FSH), luteinizing hormone (LH), estradiol (E(2)) and progesterone (P(4)) from 30 workers respectively in the exposure group and the control group selected randomly (in periovulatory period) were detected with the radio immunoassays.</p><p><b>RESULTS</b>With the increase of residue of HF-EMF, the incidence of menstrual disorder was increased (33.8% and 26.8% in the exposure group compared with 12.0% in the control group) (P < 0.01) and the incidence of menorrhagia in the exposure I group (16.9%) was significantly higher than that in the control group (8.0%) (P < 0.01). However, serum P(4) was significantly lower in the exposure group [(2.10 +/- 1.57) and (2.39 +/- 1.35) mg/L] than in the control group [(3.80 +/- 2.67) mg/L] (P < 0.01). There was no significant difference in serum E(2), LH and FSH between the exposure and the control groups.</p><p><b>CONCLUSION</b>Increase of the incidence of menstrual disorder as well as that of menorrhagia, and decrease of levels of some reproductive hormones are closely associated with HF-EMF (25 approximately 30 MHz).</p>
Subject(s)
Adolescent , Adult , Female , Humans , Young Adult , Case-Control Studies , Electromagnetic Fields , Endocrine System , Radiation Effects , Hormones , Blood , Menstruation Disturbances , Epidemiology , Occupational Exposure , Surveys and QuestionnairesABSTRACT
OBJECTIVE@#To explore social function in assessment of self-defense capability of patients with mental retardation against sexual assault.@*METHODS@#The data of 125 sexual assault victims with mental retardation were collected according to our forensic psychiatry assessment criteria. Mental retardation was rated with WAIS-RC, GAF, and the rating scale of the intelligence disability.@*RESULTS@#There were significant differences in occupation, marital status, education, knowledge about sex, and history of pregnancy among the patients with different levels of self-defense capability. The kappa values of the rating scale score of the intelligence disability, GAF, and IQ were 0.710, 0.661, and 0.494; the kappa values of that and self-defense capability were 0.538, 0.472, and 0.316 respectively.@*CONCLUSION@#The level of social function is very important in the assessment of self-defense capability in patients with mental retardation.
Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Crime Victims , Forensic Psychiatry/methods , Intellectual Disability/diagnosis , Intelligence , Intelligence Tests , Retrospective Studies , Severity of Illness Index , Sex Offenses , Social AdjustmentABSTRACT
<p><b>AIM</b>To investigate the effect on myocardial apoptosis and Bcl-2/Bax induced by remote preconditioning (RP) and to discuss the hypothesis from opioid receptors in pigs.</p><p><b>METHODS</b>Skeletal muscle ischemia was performed in pigs by occlusion of the femoral artery (FAO) for 15 min followed by a 10 min of reperfusion. Infarction of the heart was induced by 40 min of left anterior descending coronary artery (LAD) occlusion followed by 120 min reperfusion. In the RP model induced by FAO, the role of opioid receptors was investigated by using antagonist of the opioid receptors (naloxone). The signal transduction pathway of RP was investigated by using hexamethonium. Apoptosis of left ventricular samples from nonischemic and ischemic areas was detected in situ with end-labeling (TUNEL) method and measured by flow cytometry. Bcl-2 and Bax was also measured by flow cytometry.</p><p><b>RESULTS</b>(1) The apoptosis rate in ischemic myocardium in RP group measured by flow cytometry was lower (4.43% +/- 0.74%) compared with that in CONT group (15.4% +/- 1.15%), but Bcl-2/Bax was higher (1.36 +/- 0.09, CONT group: 0.56 +/- 0.08). (2) The protective effect could be prevented by naloxone used before RP protocol (apoptosis rate: 13.0% +/- 0.56% and Bcl-2/Bax: 0.69 +/- 0.18, P < 0.05). (3) Naloxone had no effect on apoptosis rate in CONT group. (4) Hexamethonium used before RP protocol had no effect on apoptosis rate and bcl-2/bax. Apoptotic cardiomyocytes detected in TUNEL correspond to the above.</p><p><b>CONCLUSION</b>RP induced by skeletal muscle ischemia could prevent myocardium from apoptosis, in which Bcl-2 and Bax might take part in regulation and control. Furthermore opioid receptors could take part in triggering the course and a neuronal signal transmission from the remote area to heart could be excluded.</p>
Subject(s)
Animals , Apoptosis , Ischemic Preconditioning , Methods , Muscle, Skeletal , Myocardial Reperfusion Injury , Pathology , Myocardium , Pathology , Receptors, Opioid , SwineABSTRACT
Objective To improve the CR image quality and to promote the digital image standard constitution by analyzing the common problems and malpractiees in application of computed radiography. Methods Phenomenon and reasons of 107 CR junk-films from nine three-"A"-hospitals were analyzed, discussed,recorded,and statistised by 20 radiologists,radiographers and engineers.Results Among 107 junk fihns,there are 36 cases(33.64%)of incorrect operations,29 cases(27.10%)of artifacts in reading and transferring the data of IP,15 cases(14.02%)of artifacts in IP system,and 13 cases (12.15%)of selection of inappropriate radiographic parameters,and 9 eases(8.41%)of printer-failures, and 5 eases(4.67%)of inappropriate post-processing techniques.By analyzing the reasons of 107 junk films we found that 60.74% were due to less responsibilities and incorrect operations,and 35.51% were due to new problems in CR techniques,and other were due to inappropriate post-processing techniques. Conclusion Responsibilities,operation regulations,digital image quality standards,studying of new techniques and appropriate use of the post-processing techniques are the key points for improving the CR image quality and the diagnosis level.