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Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.
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Objective To investigate the level change of interleukin-37(IL-37) and its clinical significance in the patients with chronic kidney disease(CKD) and nephrotic syndrome(NS) .Methods 57 cases of CKD (CKD group) and 13 cases of NS (NS group) in the Affiliated Donghua Hospital of Sun Yat-sen University from September to November 2016 were selected .The CKD group was redivided into the CKD stage 1 ,stage 3 and stage 5 groups .At the same time 22 individuals undergoing physical examina-tion were selected as the healthy control group .The ELISA sandwich method was adopted to detect the plasma IL-37 level .Then the detection results were statistically analyzed .Results The level of plasma IL-37 in the CKD group and NS group was significant-ly higher than that in the healthy control group ,the difference was statistically significant (P<0 .01);the level of plasma IL-37 had no statistical difference among the CKD stage 1 group ,CKD stage 3 group and CKD stage 5 group(P>0 .05);the plasma IL-37 lev-el after treatment in the CKD group and NS group was decreased compared with before treatment ,the difference was statistically significant(P<0 .01);the plasma IL-37 level in CKD patients was positively correlated with the WBC and lymphocyte cells levels . Conclusion The patients with CKD and NS have high level expression of plasma IL-37 ,which might have good auxiliary diagnostic value for CKD and NS .
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We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.
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Objective:To establish a double antibody sandwich ELISA assay for detection of human IL-37 in serum.Methods: Mouse anti-human IL-37 monoclonal antibody was used as capturing antibody,rabbit anti-human IL-37 polyclonal antibodies served as detection antibody,HRP labeled goat anti-rabbit IgG employed as second antibody and recombinant human IL-37 protein used as reference standard for the establishment of a doubleantibody sandwich ELISA.The working conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients with Dengue fever.Results: The sensitivity of the established ELISA method was 1.465 μg/L approximately.Likewise,the linearity range of this method was about (1.465-46.875) μg/L.Further,the co efficient of variation (CV) of inter-batch and intra-batch in this study were 6.6％ and 11.7％,respectively.Notably,this method could be used in the detection of IL-37 in serum of the patients with Dengue fever,showing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls.Conclusion: The double antibody sandwich ELISA assay for the detection of human IL-37 was successfully established,which can be apply to detect of human IL-37 in clinical samples.
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Objective To analyze the dynamic changes of white blood cells in patients with cavitary pulmonary tuberculosis (CPTB) during chemotherapy procedure ,and explore immunological indexes which could evaluate the curative effect and prognosis . Methods 85 cases of patients with CPTB (CPTB group) and 100 healthy people conducted physical examination (control group) were enrolled in this study ,The immunological indexes of white blood cells were detected and compared between CPTB group and control group ,and dynamic changes of white blood cells were analyzed in patients with CPTB .Results Compared with the control group ,absolute lymphocyte count and percentage of lymphocyte were decreased in the CPTB group before chemotherapy ,while the total white blood cells count ,absolute neutrophil count and percentage of neutrophil ,absolute monocyte count and percentage of monocyte were increased in the CPTB group before chemotherapy ,there were statistically significant differences between the two groups(P<0 .05) .Except the absolute lymphocyte count ,other indexes were statistically increased or decreased during chemothera‐py procedure .Conclusion After chemotherapy ,the immunological reactions induced by lymphocyte are under inhibitory state .The dynamic changes of white blood cells could reflect the state of cellular immunity in patients with CPTB during chemotherapy ,which could be helpful for the evaluation of curative effect and prognosis .
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Objective To detect plasma interleukins-35 (IL-35 )level in the patients with active tuberculosis complicated with bronchiectasis and to analyze its clinical significance.Methods Peripheral blood of patients with active tuberculosis from depart-ment of Dongguan 6th People′s hospital were collected,assigned to the active tuberculosis complicated with bronchiectasis group and active tuberculosis group.The healthy volunteers served as the control group.The plasma IL-35 level was measured by ELISA, and peripheral blood neutrophils and lymphocytes were detected by hematology analyzer.Results The levels of plasma IL-35 signif-icantly increased in both patients with active tuberculosis complicated with bronchiectasis and patients with active tuberculosis.The level of plasma IL-35 of patients with active tuberculosis complicated with bronchiectasis was significantly higher than that of the patients with active tuberculosis.The absolute value and percentage of peripheral blood neutrophils of patients with active tubercu-losis complicated with bronchiectasis were significantly higher than those of healthy volunteers.However,the percentage of periph-eral blood lymphocytes of patients with active tuberculosis complicated with bronchiectasis was significantly lower than that of healthy volunteers.Pearson correlation analysis showed that the absolute value of peripheral blood neutrophils of patients with ac-tive tuberculosis was positively correlated to the level of plasma IL-35.Conclusion IL-35 may play an important role in the progres-sion of active tuberculosis complicated with bronchiectasis.The determination of IL-35 may be helpful to the diagnosis of patients with active tuberculosis complicated with bronchiectasis.
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AIM:To investigate the effect of mesalazine treatment on regulation of Th1, Th17 and Treg cells in mice with dextran sulfate sodium ( DSS)-induced ulcerative colitis ( UC) .METHODS:The expression of IL-17, IFN-gam-ma and Foxp3 in the peripheral blood mononuclear cells ( PBMC) and intestinal mucosa lamina propria mononuclear cells ( LPMC) of DSS-induced UC mice was detected by flow cytometry analysis.The effect of mesalazine treatment on regulaiton of Th1, Th17 and Treg cells in the mice with DSS-induced ulcerative colitis was examined.RESULTS:The expression of IL-17, IFN-γand Foxp3 on CD4 +T cells were significantly higher in the PBMC of DSS-induced mice than those in control group.CD4+IFN-γ+T cells and CD4 +Foxp3 +T cells were higher in LPMC than those in control group, except CD4+IL-17 +T cells.Moreover, the Th1, Th17 and Treg cells were higher in DSS group than those in control group in LPMC.How-ever, only Tregs was higher in PBMC.Pre-treatment with mesalazine significantly decreased the number of Th17, Th1 and Treg cells of UC model mice both in PBMC and LPMC.CONCLUSION: The Th1, Th17 and Tregs cells in DSS-induced mice were significantly higher than those in control mice, suggesting that CD4 +T cell subsets play an important role in the pathogenesis of UC.Mesalazine may play a role in the treatment of UC by regulating the Th1, Th17 and Tregs cells.
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To adapt to the system reformation of medical laboratory science from five academic years to four academic years and to meet the new professional technology-oriented requirements, the medical laboratory science institute of Guangdong Medical College has carried out a comprehensive reform of curriculum system. This paper has analyzed the current problems in the school medical ex-amination and explored the curriculum system reform from three respects such as adjusting curriculum by restructuring and integrating programs, implementing modular teaching to build its characteristics and strengthening practice teaching.And it has also explored the full assessment mode by optimizing the traditional one-stop assessment.
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Stromal cell types of the neoplastic microenvironment play important roles in the occurrence and development of tumor.Tumor-associated fibroblasts (TAFs) are the most abundant cells in tumor stroma.They promote the malignant transformation of epithelial cells and other cells through cell-cell communication via various soluble factors.
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Objective To investigate the risk factors related to progressive intracranial hemorrhage (PIH) in patients with acute traumatic brain injury (TBI) and analyze their clinical significance.Methods PIH was validated by comparing the initial and repeated CT scans. Data including gender,age, injury causes, Glasgow Coma Score (GCS) on admission, time interval from injury to the first CT scan, initial CT scan manifestations, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fg), thrombin time (TT), platelet (PLT) and D-dimer (D-D) in both groups were compared with Logistic regression analysis to observe the risk factors related to PIH. Results The study involved 498 patients with acute TBI, of which 139 (27.91%) patients suffered from PIH. There were 116 patients (83.45%) with PIH who received the initial CT scan within two hours post injury.There was statistical difference in aspects of age, GCS on admission, time interval from injury to the first CT scan, initial CT scan manifestations ( including fractures, subarachnoid hematoma, contusion and onset hematoma), PT, Fg and D-D values in both groups (P ＜0.01 ). Logistic regression analysis showed that CT scans (subarachnoid hemorrhage, brain contusion and primary hematoma) and plasma D-D values were predictors of PIH ( P ＜ 0.01 ). Conclusions For patients with the initial CT scan manifestations including subarachnoid hemorrhage, brain contusion, primary hematoma together with D-D value increase within two hours post injury, a continuous CT scan should be performed promptly to detect PIH early.
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Objective To explore the effects and mechanism in patients with ulcerative colitis (UC) treated by Yin Yang equilibrated preparation. Method 82 patients with UC as treatment group and 35 health parsons as control group underwent the testing. The serum and mucosal of all testing persons were taken pre and post treatment to detect CD+4 CD+25 regulatory T cells(Tregs) by flow cytometry at the end of one month,treating with SASP and Yin Yang equilibrated preparation. Result Compared with normal control group,the proportion of CD+4 CD+25 Tregs was markedly decreased in PB and mucesal of group A and B(P <0.01). But it was significanfly increased in therapeutic groups of SASP and Yin Yang equilibrated preparation, and their CD+4 CD+25 Tregs in PB and mucosal were much more than the control group at the end of one month after treating with SASP and Yin Yang equilibrated preparation(P <0.01 or P< 0.05). Conclusion CD+4 CD+25 Tregs with strong immune suppression play a central role in the initia-tion and development of UC,and the Yin Yang equilibrated preparation might exert its therapeutic effects on UC by the regulation of number and function of CD+4 CD+25 Tregs.
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Objective:To observe the effects of sera Th1/Th2-typed cytokines in patients allergic rhinitis (AR) who were treated with kebimin decoction(KD),and to study the therapeutic mechanism of KD in AR.Methods:60 patients of AR were randomly divided into two groups (treating group,TG and control group,CG). Patients in TG were treated with KD,while those in CG were treated with Xinfang Rhinitis Capsule before and after treatment,the levels of serum Th1-typed cytokines IFN-?、IL-2、IL-12 and Th2-typed cytokines IL-4、IL-5、IL-10 were determined by enzyme-linked immune sorbent assay (ELISA); and compared with 30 healthy controls.Results:The levels of serum Th2-typed cytokines IL-4、IL-5、IL-10 were higher and the levels of Th1-typed cytokines IFN-?、IL-2、IL-12 were lower in AR than that in healthy control ( P 05).Conclusion:Kebimin decoction is highly effective in treating allergic rhinitis by regulating the levels of Th1、Th2-typed cytokines,correcting the imbalance Th1/Th2-typed cytokines network.