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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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AIM:To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentia-tion of K562 cells.METHODS:The K562 cells were treated with different concentrations of DAC .The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture .The cell viability was detected with MTT assay.The morphologic features were observed under inverted microscope with Wright ’s staining.The changes of the cell cycle distribution and the expression of CD 11b and CD42b were analyzed with flow cytometry .The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot .RESULTS:DAC signifi-cantly decreased the colony number of the cells and cell viability in a dose-dependent manner .The morphological changes of the cells displayed partial differentiation .After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased , while the cell proportion in G 2/M phase was obviously increased in a dose-dependent manner . After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated , and the protein expression of P27, GATA-1 and PU.1 was increased.However, the protein expression of CDK2 and cyclin E1 was decreased .CONCLUSION:DAC inhibits the proliferation and induces differentiation of the K 562 cells via regulation of cell cycle .
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Objective To investigate the distribution and antibiotic resistance of clinical Pseudomonas aeruginosa in Yunnan provincal of China in 2014.Methods Pseudomonas aeruginosa were collected from 28 hospitals in Yunnan surveillance of China.All hospitals were carried with the unified solution for bacteria culture,isolation,identification and antibiotic sensitivity tests according to CLSI M100-S24.The data of 2 873 strains pseudomonas aeruginosa were analyzed by WHONET5.6 software.Results 2 873 clinical strains of non-repetitive Pseudomonas aeruginosa isolates,90.36%were isolated from hospitalized patients and 60.32% from sputum,8.42% from urine,8.11% from secretion,4.70% from abscess,2.92% from blood,etc.The sensitive rates of common antimicrobial agents of Pseudomonas aeruginosa in top five were turn amikacin (88.7%),piperacillin/he azole temple (85.0%),tobramycin (83.1%),piperacillin (80.3%) and cefepime (80.1%).59.0% of the Pseudomonas aeruginosa strains were resistant to Aztreonam.20.9%-29.7% of the Pseudomonas aeruginosa strains were resistant to Imipenem,Ceftazidime,Meropenem,Ciprofloxacin and Levofloxacin.The Pseudomonas aeruginosa isolates showed the lowest resistance rate (10.3%-19.9%) to Piperacillin,Gentamicin,Cefepime,Tobramycin,Piperacillin/Tazobactam and Amikacin.Conclusion The antimicrobial susceptibility pattern varies widly with Pseudomonas aeruginosa isolated in Yunnan of China in 2014.Antimicrobial resistance sur-Monitoring the antibiotic resistant trend of Pseudomonas aeruginosa and implementing the nosocomial infection control policy become more important in hospital management setting.
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Objective To study the incidence and clinical significance of abnormal neuroendocrine hormone in patients with the syndrome of brain injury.Methods 67 cases with the syndrome of brain injury were included in the study group,and 95 cases without the post traumatic syndrome after brain injury were included in the control group.The level of FT3,FT4,TSH,growth hormone(GH),andrenocortico hormone(ACTH),cortisol (Cor),prolactin(PRL),testosterone (T),estradiol (E2),follicular stimulating hormone (FSH),luteotropic hormone (LH),and progesterone (P) in peripheral blood were measured by radioimmunoassay.The incidence of the abnormal neuroendocrine hormone after brain injury was statistically analyzed.Patients with abnormal hormone were given hormone replacement therapy and the curative effects were observed.Results The incidence of neuroendocrine hormone abnormalities was 38.8％ in patients with the syndrome of brain injury,while it was 10.5％ in the control group.There was significant difference between the 2 groups(P＜0.05).The symptom remission rate was 88.5％ after hormone treatment.Conclusions There was a high incidence of abnormal neuroendocrine hormone secretion in patients with the syndrome of brain injury.The hormone level may be used as an important index to guide clinical therapy.
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Aim To observe mesenchymal stem cell differentiation into osteoblast induced by TSPG, and explore how TSPG enhances the promotion of hemato-poiesis of osteoblast differentiation from mesenchymal stem cell. Methods MSCs were cultured by TSPG combined with osteoinductive medium. The cellular vi-ability of proliferation was detected with MTT assay. The content of alkaline phosphatase in the cultural su-pernatant was tested with pNPP assay. The ability of MSCs to form calcium nodes was also observed after a-lizarin red stain. The protein expression of RUNX2 was analyzed with Western blot. The content of cytokines associated with hematopoiesis was tested with Elisa as-say. The ability of promoting hematopoiesis was detec-ted with hematopoietic colony forming assay. Results Both MTT and pNPP assay showed that optical den-sity ( OD) values were increased in response to TSPG treatment in a dose-dependent manner. The mineraliza-tion formation ability was enhanced with TSPG-treat-ment. Meanwhile, the expression of RUNX2 protein was up-regulated in TSPG-treated cell. Moreover, the content of cytokines associated with hematopoiesis and the number of hematopoietic progenitor colony were in-creased by TSPG-treatment compared with the control group. Conclusion TSPG could induce MSCs differ-entiation in to osteoblast and enhance the effect of oste-oblast differentiation from MSCs on promoting hemato-poiesis.
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Objective To learn the species distribution characteristics and proportion occurrence of methicillin-resistant strains about Staphylococcus detected in the First People′s Hospital of Kunming.Methods The species distribution characteristics and proportion occurrence of methicillin-resistant strains were analyzed retrospectively from January 2005 to December 2013.Results A total of 3 561 Staphylococcus strains were detected in 9 years,included 21 species and subspecies,and another 12 strains were not i-dentified to species.2005-2013 species composition showed an increasing trend,there were five kinds of Staphylococcus in 2005, until 2013 reach to 13 kinds.Each year the main bacterial were Staphylococcus aureus,Staphylococcus epidermidis,Staphylococcus haemolyticus and staphylococcal hominis.Methicillin resistant Staphylococcus aureus incidence decreased significantly since 201 1, decrease from 76.3% in 2010 to 25.6% in 2013.Staphylococcus epidermidis,Staphylococcus haemolyticus,Staphylococcal hominis and coagulase-negative Staphylococci resistant to high incidence of methicillin-resistant strains of the average,remained stable at a-round 70.0%.Conclusion The distribution characteristic of Staphylococcus in this hospital was increasingly complex year by year, the opportunity of infection caused by Staphylococcus was also increased,the detection rate of methicillin-resistant strains was high, it should be noted to use clinical drug rationally.
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[ ABSTRACT] AIM:To observe the effects of total saponins of Panax ginseng ( TSPG) on the promotion of hema-topoiesis and to explore the underlying mechanism in treating aplastic anemia ( AA) in a mouse model.METHODS:For preparation of AA model, BALB/c mice were exposed to sublethal dose of 5.0 Gyγradiation, followed by transplantation of lymphocytes from DBA/2 donor mice.The experiment was divided into 6 groups, including normal control group, AA model group, TSPG treatment groups with low, medium and high doses, and positive control group with cyclosporine A ( CsA) .Both TSPG and CsA were administered by gastrogavage.After 15 d treatment, the peripheral hemogram was test-ed, the cytokine contents in serum were measured, and bone marrow semisolid culture of colony-forming assay was conduc-ted for determining hematopoietic progenitor cells.The protein levels of extracellular signal-regulated kinase 1/2 ( ERK1/2) and its phosphorylation status in the bone marrow cells were determined.RESULTS:Curative effect of TSPG in trea-ting AA mice was satisfactory.Peripheral counts of white blood cells and platelet, and concentration of hemoglobin in TSPG groups with medium and high doses were significantly higher than those in model control.TSPG increased the colony num-bers of CFU-E, BFU-E, CFU-GM and CFU-MK derived from hematopoietic progenitor cells.Meanwhile, up-regulation of the phosphorylated ERK1/2, decreased contents of Th1 cytokines and increased contents of Th2 cytokines in the serum were observed.CONCLUSION:TSPG possess the dual efficacies on the promotion of hematopoiesis and immunoregulation in AA mice by regulating the expression of Th1/Th2/Th17 cytokines and increasing the phosphorylation of ERK1/2.
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Objective To analyze the detected pathogens composition in positive blood culture samples and drug resistance in our hospital from January 2005 to December 2012 in order to accumulate the data information of pathogenic bacteria distribution and drug resistance in bacteremia .Methods The BD9240 and BacT /Alert3D 240 blood culture systems were used to perform the blood culture .The identification of isolated bacteria and the drug susceptibility test were conducted by using Microscan walkaway 40 sys‐tem and the Vitec2 compact system .The Data were analyzed by adopting the Whonet5 .6 software .Results In 1 829 positive bacte‐rial strains by blood culture ,986 strains were Gram negative bacilli ,accounting for 53 .9% ;721 strains were Gram positive coccus , accounting for 39 .4% ;104 strains were fungi ,accounting for 5 .68% .The resistant rate of staphylococcus to vancomycin ,linezolid and teicoplanin was 0% ,which to amoxycillin/clavulanic acid ,rifampicin ,amikacin ,sulfamethoxazole compound and chloramphenicol was lower than 40% .The sensitive of enterococcus to linezolid and teicoplanin was 100% ,but enterococcus faecium was resistant to vancomycin(2 .6% ) .The penicillin resistant rate of Streptococcus was 21 .7% .The resistant rates of E .coli and K lebsiella pneumo‐nia were 0% to imipenem and meropenem ,and less than 22% to amikacin ,piperacillin/tazobactam and cefoxitin .The resistant rates of salmonella to CLSI recommended five kinds of detection drug were less than 6 .5% .The resistant rates of pseudomonas aerugino‐sa were more than 25% to imipenem and more than 25% to meropenem .Conclusion The pathogens spectrum detected by blood culture is widespread .The resistance rates of different bacteria vary widely .
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Aim To observe the effects of panaxdiol saponins component ( PDS-C) extracted and isolated from Chinese ginseng herb as new Chinese patent med-icine on the promotion of hematopoiesis and the regula-tion of the immune system in treating mice models with aplastic anemia ( AA ) . Methods For preparation of immune mediated AA models, BALB/c mice were ex-posed to sublethal doses of 5. 0 Gy γ radiation, fol-lowed by transplanted lymphocytes from DBA/2 donor mice. The mice models were divided into six groups in-cluding normal control, AA model, PDS-C treated groups with lower, medium and higher dosages, cy-closporine ( CsA) as positive drug control. Both PDS-C and CsA were administered by gastrogavage for 15 days. The peripheral blood cells counts and bone mar-row pathological examination were tested, the percenta-ges of Th1/Th2/Treg cells from spleen were measured, the protein expression levels of T-bet, GATA-3 and FOXP3 transcription factors in spleen cells were detec-ted. Results Curative effect of PDS-C on treating AA mice was satisfactory. The peripheral hemoglobin, white blood cells and platelet counts in PDS-C groups with medium and higher doses were significantly higher than those in model control. Meanwhile, PDS-C ele-vated the percentages of Th2 cells and Treg cells, but decreased the percentage of Th1 cells, as well as up-regulated the GATA-3 , FOXP3 and down-regulated the T-bet protein levels. Conclusion PDS-C possesses the activities of promoting hematopoiesis obviously. It can improve marrow myelosuppression, enhance the re-covery of hematopoiestic function, and elevate the pe-ripheral blood cells counts. PDS-C also pays its immu-noregulatory efficacy though recovering from unbal-anced Th1/Th2/Treg cells in treating immune media-ted AA mice.
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Aim To investigate the effect of resveratrol on proliferation and differentiation in K562 cells. Methods K562 cells were treated with different con-centrations of resveratrol for 6d. The colony number of K562 cells was detected with semi-solid culture assay. Expression of GATA-1 and PU. 1 in K562 cells was re-spectively measured with immunocytochemistry and Western blot. Expression of differentiation related anti-gen, CD11b, CD14 and CD42b, was measured with flowcytometry on K562 cells. Results Resveratrol could significantly decrease the colony number of K562 cells in a dose-dependent manner, and enhance the ex-pression of GATA-1,PU. 1,CD11b, CD14 and CD42b in K562 cells. Conclusion Resveratrol could inhibit the proliferation and induce differentiation of K562 cells via up-regulating the expression of GATA-1 and PU. 1 protein.
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Objective To observe the effects of whole body γ-ray radiation on hematopoiesis and cytokines related to T cell subsets in mouse,to detect the expression of transcription factors of splenic T cell subsets,and to investigate the correlation between hematopoiesis injury and abnormal immune function.Methods Totally 50 BALB/c mice were divided into radiation group and blank control group with the random number table method.The former group were given 5.5 Gy 60Co γ-ray radiation on whole body and another received sham radiation.The numbers of white blood cells and platelets of radiation group were counted at 4,8,12 and 20 d after radiation,and these numbers of blank control mice were counted only at 20 d.Hematopoietic tissue proliferation was evaluated by biopsy sections of mice femur.The contents of Th1,Th2,and Th17 in peripheral blood were detected with cytometric bead array (CBA).The expressions of T-bet/GATA-3 and RORγt/Foxp3 proteins related to the differentiation of T cell subsets in spleen tissue were measured by Western blot.Results The numbers of white blood cells and platelets of radiation group mice were reduced obviously (t WBC =18.48,15.72,9.79,3.30; t PLT =22.52,19.74,11.78,4.70,P ＜ 0.05) compared with blank control group.Biopsy sections showed that bone marrow hematopoietic cells of the radiated mice were less than those of blank group,and adipocytes became more.At 8 d,the marrow suppressions were more obvious than those at 20 d.Serum contents of Th1 cytokines IFN-γ,TNF-α and Th2 cytokines IL-4,IL-6 in the radiation group were higher than those in the blank control group at 8 and 12 d(t IFN-γ =2.93,3.36,t TNF-α =6.09,8.11,6.43,4.49,tIL-4 =4.49,3.18,t IL-6=5.11,8.67,6.67,8.55,P＜0.05).IL-17A secreted mainly by Th17 cells was also higher than the blank (t =3.68,6.24,5.32,4.06,P ＜ 0.05).Compared with the blank control group,the expression of T-bet protein increased significantly (t =5.64,2.75,3.56,4.65,P ＜ 0.05),and the expressions of GATA-3,RORγt,and Foxp3 proteins decreased at 4,8 and 12 d except the RORγt at 20 d (tRORγt =6.79,4.31,4.47,tGATA-3 =3.88,8.06,2.84,3.23,tFoxp3 =10.00,8.06,2.89,5.93,P＜ 0.05).Conclusions 5.5 Gy whole body γ-ray radiation inhibits bone marrow hematopoiesis of BALB/c mice and makes the differentiation and function of T cells to be abnormal,which may be associated with bone marrow hematopoiesis obstacle.
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Objective: To construct the packaging plasmid pSNAV-hEndostatin-CMV-EGFP of adeno-associated virus vector (AAV) encoding human Endostatin (hEndostatin) cDNA. Methods: The hEndostatin eDNA obtained from plasmid pCD-sEndostatin was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hEndostatin-CMV-EGFP was identified by PCR analysis, restriction enzymes analysis and sequencing analysis.Results: The recombinant pSNAV-hEndostatin-CMV-EGFP was correctly constructed and confirmed by PCR analysis, restriction enzymes analysis and sequencing analysis. Conclusion: The constructed AAV-hEndostatin packaging plasmid pSNAV-hEndostatin-CMV-EGFP could be the packaging plasmid of rAAV-hEndostatin-EGFP.
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[Objective]To explore the potential alteration of their osteogenesis and adipocyte differentiation in bone marrow mesenchymal stems cells from mice with immune-mediated aplastic anemia.[Methods]Balb/c mice model of immune-mediated aplastic anemia was established by radiation with sublethal dose of 60Co following the intravenously infusion lymphocytes of DAB/2 mice.The culture of the MSC and CFU-F and pathological examination of bone marrow were carried out 15 days later.The amount of calcium node and the frequency of adipocyte differentiation were evaluated respectively by alizarin red and oil red O.[Result]The number of CFU-F and the number of calcium node in model mice decreased more greatly than normal mice,but the ferenqucy of adipocyte differentiation was more increased greatly in model mice than normal mice;the pathological examination showed in the model mice,the hematopoietic structure was destroyed and filled with abundant adipocyte.[Conclusion]The potention of osteogenesis and adipocyte differentiation was altered in the mice with immune-mediated aplastic anemia.
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Objective To explore the effects of garlic and oat and their compounds on lipid metabolism and hemorheology of hyperlipidemic rats. Methods Sixty adult Wistar rats were randomly divided into five groups (12 in each group), and fed in different cages with basic forage, hyperlipid forage, low garlicin forage, high garlicin forage, and compound forage of garlicin and oat for 8 weeks. The blood lipid and hemorheology of hyperlipidemic rats were measured. Results After the rats were fed with low garlicin forage, high garlicin forage, and compound forage for 8 weeks, the levels of TC, TG, VLDL, and LDL decreased significantly, but HDL increased significantly. The contents of the high, middle and low blood viscosity, and plasma adhesion, blood reduction viscosity and hematocrit decreased significantly in low garlicin forage group, high garlicin forage group, compound forage of garlicin and oat group. In the normal forage group, these changes were not significant. In hyperlipid forage group, the contents of serum TG and TC, plasma VLDL-C and LDL-C increased significantly, but the contents of plasma HDL-C decreased significantly. Conclusion Garlic and oat may play a role in the decrease of lipid and ameliorate hemorheology, but low-dose compound forage of garlicin and oat may facilitate lipid metabolism and prevent hyperliemia and atherosclerosis.