1.Construction and biological characterization of lmo2363 gene deletion strain of Listeria monocytogenes
Ji ZHI ; Qing CAO ; Xuehui ZHAO ; Haohao ZHANG ; Ziqiu FAN ; Yonghui MA ; Jing DENG ; Zengwen HE ; Jinrui MA ; Kunzhong ZHANG ; Qian CHONG ; Caixia WANG ; Huiwen XUE ; Huitian GOU
Chinese Journal of Veterinary Science 2024;44(9):1923-1929,1956
This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7%NaCl,10%NaCl,3.5%ethanol,4.0%ethanol and pH5 stress(P<0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P<0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P<0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.
2.Analysis of medical reimbursement rate and influencing factors under the DIP payment method
Meng-Yuan ZHAO ; Kun-He LIN ; Ying-Bei XIONG ; Yi-Fan YAO ; Zhi-He CHEN ; Yu-Meng ZHANG ; Li XIANG
Chinese Journal of Health Policy 2024;17(6):40-46
Objective:Analyze the medical reimbursement rate and influencing factors under the DIP payment method to refine the DIP payment policy,promote the optimization of internal operations in medical institutions,and ensure reasonable compensation.Methods:Based on the 2022 DIP fund settlement data from 196 medical institutions in City A,the study used multiple linear regression to analyze the factors affecting medical reimbursement rate and conducted a heterogeneity analysis for medical institutions of different levels.Results:The medical reimbursement rate for medical institutions in City A in 2022 was 103.32%.Medical institutions with lower CMI standardized inpatient costs,lower rates of deviation cases,tertiary care institutions,lower proportion of level-four surgeries,and lower ratios of resident to employee medical insurance cases have higher medical reimbursement rate(P<0.05).Heterogeneity analysis reveals that therates of deviation cases,the proportion of primary care diseases,the ratio of resident to employee medical insurance cases,and the low-standard admission rate have different impacts on medical institutions of different levels.Conclusion:Medical insurance departments should improve policies for primary care diseases,dynamically adjust disease catalogs and payment standards,optimize funding levels and institutional coefficients,and increase penalties for violations to ensure effective use of funds.Medical institutions need to strengthen their understanding of policies,focus on refined internal management,promote standardized and rational diagnosis and treatment through performance assessment transformation,and leverage their own advantages in medical services to reasonably increase the medical reimbursement rate.
3.Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines
Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG
Parasites, Hosts and Diseases 2024;62(4):476-483
Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines.
4.Protective effects of Shiwei Ruxiang Powder on renal injury in rats with gouty nephritis by regulating mitochondrial autophagy
Yan-Rong ZHU ; He-Bing XIE ; Chun-Xiang GONG ; Jie-Nan ZHAO ; Zhi-Bing YUAN
Chinese Traditional Patent Medicine 2024;46(9):2923-2930
AIM To investigate the renal protective effects of Shiwei Ruxiang Powder on gouty nephritis in rats based on mitophagy.METHODS Rats were randomly divided into the blank group,the model group,the low-dose,medium-dose,and high-dose Shiwei Ruxiang Powder groups(200,400,800 mg/kg)and allopurinol group(10 mg/kg).The rat model of gouty nephropathy was established by gavage of potassium oxyzinate(750 mg/kg)and uric acid(300 mg/kg).The rats had their levels of UA,SCr,BUN,XOD,SOD,MDA,ROS measured by automatic biochemical analyzer,ELISA and chemical fluorescence method;their renal pathological changes observed by HE staining;their apoptosis of renal tissue cells observed by TUNEL staining;and their mRNA and protein expressions of IL-1β,TNF-α,Bax,Bcl-2,caspase-3,caspase-9,PINK1,Parkin and LC3-Ⅱ detected by RT-qPCR and Western blot.RESULTS Compared with the model group,Shiwei Ruxiang Powder groups displayed dose-dependently decreased serum levels of UA,BUN and SCr,renal deposition of urate crystal and apoptosis(P<0.05);decreased renal levels of ROS and inflammatory factors IL-1β and TNF-α(P<0.05);and increased renal expressions of mitochondrial autophagy-related proteins PINK1,Parkin and LC3-Ⅱ(P<0.01).CONCLUSION Shiwei Ruxiang Powder may relieve gouty kidney injury in rats by reducing the uric acid level,the renal oxidative stress and inflammatory response,and activating mitophagy pathway as well.
5.Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines
Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG
Parasites, Hosts and Diseases 2024;62(4):476-483
Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines.
6.Allyl isothiocyanate exacerbates acute toxoplasmosis through inhibition of inflammatory cytokines
Qiu-Mei LIN ; Hong-Bin LONG ; Jun-Ting HE ; Zhi-hao ZHANG ; Ho-Woo NAM ; Fu-Shi QUAN ; Qi ZHONG ; Xu-Qing LIU ; Zhao-Shou YANG
Parasites, Hosts and Diseases 2024;62(4):476-483
Allyl isothiocyanate (AITC) is a natural product commonly used in food preservation and pharmaceutical applications. Toxoplasmosis, caused by the protozoan pathogen Toxoplasma gondii, is prevalent globally while the impact of AITC on toxoplasmosis is unclear. We explored the effect of AITC on acute toxoplasmosis. We infected C57BL/6 mice with T. gondii type I RH strain following AITC administration. On the 4th day after infection, which corresponds to the initial stage of infection, we collected serum for the determination of inflammatory cytokine levels. The mice serum of the AITC-administered group contained significantly lower levels of granulocyte colony-stimulating factor, interferon-gamma, interleukin (IL)-23 subunit p19, IL-4, IL-6, and monocyte chemoattractant protein-1. The lifespan of the mice in the AITC-administered group was significantly reduced. In vitro experiments showed that AITC promoted the proliferation of intracellular T. gondii accompanied by the inhibition of IL-4, IL-1β, and IL-6 production in RAW264.7 macrophages. Our results showed that AITC facilitated T. gondii infection in the early stage by inhibiting the production of several inflammatory cytokines.
7.Rapid diagnosis of invasive candidiasis by droplet digital PCR
Zhijie HE ; Weichao LI ; Minghui HE ; Xiaotong CHEN ; Zhao LIN ; Yaowei ZHI
The Journal of Practical Medicine 2024;40(19):2738-2746
Objective To establish a rapid detection method for invasive candidiasis based on droplet digital polymerase chain reaction(ddPCR).Methods We developed an assay system using a microtitre-based digital PCR platform and designed primer probes specific for four Candida species,namely Candida albicans,Candida smoothii,Candida near-smoothii,and Candida tropicalis.(1)The Limit of Blank(LOB)range and positive judg-ment value were determined by analyzing No Template Control(NTC)samples.(2)The Limit of Detection(LOD)range was determined by diluting positive samples with 10 replicate extractions at each concentration gradient.(3)The Linear Limit of Quantitation(LOQ)range was determined by repetitive testing of diluted samples.(4)The linear range limit was determined through gradient dilution of the positive samples.(5)The coefficient of variation(CV),calculated from the logarithmic values of the resultant concentrations,was assessed by extracting and test-ing positive samples in 12 repetitions at both high and low concentrations.(6)Method reliability was evaluated by calculating the CV from the logarithmic values of the resultant concentrations obtained from clinical samples with fungal culture results.Results The ddPCR assay detected Candida LOB at a range of 0~81 copies/mL,with a positive threshold set at≥3 positive microdroplets.The LOD and LOQ were determined to be 3×102 copies/mL.The linear range for detecting different concentration gradients was found to be between 3×102 and 3×107 copies/mL,with high correlation coefficients observed for Candida albicans(R2=0.999 5),Candida smoothii(R2=0.998 9),Candida near-smoothii(R2=0.999 4),and Candida tropicalis(R2=0.999).Additionally,the coefficient of variation for the resultant concentration logarithmic values was less than 5%,meeting precision requirements.Furthermore,preliminary validation using clinical specimens demonstrated consistent results compared to clinical culture findings.Conclusion ddPCR exhibits rapidity,high sensitivity,good repeatability,and high specificity in detecting inva-sive candidiasis in critically ill patients.This study highlights the potential value of droplet digital PCR as a diag-nostic tool for invasive candidiasis.
8.Risk factors for bronchopulmonary dysplasia in twin preterm infants:a multicenter study
Yu-Wei FAN ; Yi-Jia ZHANG ; He-Mei WEN ; Hong YAN ; Wei SHEN ; Yue-Qin DING ; Yun-Feng LONG ; Zhi-Gang ZHANG ; Gui-Fang LI ; Hong JIANG ; Hong-Ping RAO ; Jian-Wu QIU ; Xian WEI ; Ya-Yu ZHANG ; Ji-Bin ZENG ; Chang-Liang ZHAO ; Wei-Peng XU ; Fan WANG ; Li YUAN ; Xiu-Fang YANG ; Wei LI ; Ni-Yang LIN ; Qian CHEN ; Chang-Shun XIA ; Xin-Qi ZHONG ; Qi-Liang CUI
Chinese Journal of Contemporary Pediatrics 2024;26(6):611-618
Objective To investigate the risk factors for bronchopulmonary dysplasia(BPD)in twin preterm infants with a gestational age of<34 weeks,and to provide a basis for early identification of BPD in twin preterm infants in clinical practice.Methods A retrospective analysis was performed for the twin preterm infants with a gestational age of<34 weeks who were admitted to 22 hospitals nationwide from January 2018 to December 2020.According to their conditions,they were divided into group A(both twins had BPD),group B(only one twin had BPD),and group C(neither twin had BPD).The risk factors for BPD in twin preterm infants were analyzed.Further analysis was conducted on group B to investigate the postnatal risk factors for BPD within twins.Results A total of 904 pairs of twins with a gestational age of<34 weeks were included in this study.The multivariate logistic regression analysis showed that compared with group C,birth weight discordance of>25%between the twins was an independent risk factor for BPD in one of the twins(OR=3.370,95%CI:1.500-7.568,P<0.05),and high gestational age at birth was a protective factor against BPD(P<0.05).The conditional logistic regression analysis of group B showed that small-for-gestational-age(SGA)birth was an independent risk factor for BPD in individual twins(OR=5.017,95%CI:1.040-24.190,P<0.05).Conclusions The development of BPD in twin preterm infants is associated with gestational age,birth weight discordance between the twins,and SGA birth.
10.No Incidence of Liver Cancer Was Observed in A Retrospective Study of Patients with Aristolochic Acid Nephropathy.
Tao SU ; Zhi-E FANG ; Yu-Ming GUO ; Chun-Yu WANG ; Jia-Bo WANG ; Dong JI ; Zhao-Fang BAI ; Li YANG ; Xiao-He XIAO
Chinese journal of integrative medicine 2024;30(2):99-106
OBJECTIVE:
To assess the risk of aristolochic acid (AA)-associated cancer in patients with AA nephropathy (AAN).
METHODS:
A retrospective study was conducted on patients diagnosed with AAN at Peking University First Hospital from January 1997 to December 2014. Long-term surveillance and follow-up data were analyzed to investigate the influence of different factors on the prevalence of cancer. The primary endpoint was the incidence of liver cancer, and the secondary endpoint was the incidence of urinary cancer during 1 year after taking AA-containing medication to 2014.
RESULTS:
A total of 337 patients diagnosed with AAN were included in this study. From the initiation of taking AA to the termination of follow-up, 39 patients were diagnosed with cancer. No cases of liver cancer were observed throughout the entire follow-up period, with urinary cancer being the predominant type (34/39, 87.17%). Logistic regression analysis showed that age, follow-up period, and diabetes were potential risk factors, however, the dosage of the drug was not significantly associated with urinary cancer.
CONCLUSIONS
No cases of liver cancer were observed at the end of follow-up. However, a high prevalence of urinary cancer was observed in AAN patients. Establishing a direct causality between AA and HCC is challenging.
Humans
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Retrospective Studies
;
Incidence
;
Carcinoma, Hepatocellular
;
Liver Neoplasms/epidemiology*
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Kidney Diseases/chemically induced*
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Aristolochic Acids/adverse effects*

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