1.New progress of refractive enhancements for residual refractive error after cataract surgery
Xiang LI ; Meixin LI ; Shuo ZHANG ; Haijuan WU ; Jinsong ZHANG ; Jing WANG
International Eye Science 2025;25(6):918-923
Cataract surgery is one of the most common ophthalmologic procedures. Advances in technology and medical policies have made it more precise. Residual refractive errors and deviation of target diopters are a main cause of dissatisfaction among patients. Refractive enhancement after cataract surgery can correct or eliminate these errors, improving patients' visual quality of life. There are multiple options for correcting residual refractive errors. The best approach depends on factors like the cause of the error, degrees of residual refractive errors, type of intraocular lens, ocular comorbidities, and patient preference. This paper summarizes the incidence and types of residual refractive errors, advancements in refractive enhancement surgeries, and provides practical solutions for clinical practice.
2.Safety and effectiveness of lecanemab in Chinese patients with early Alzheimer's disease: Evidence from a multidimensional real-world study.
Wenyan KANG ; Chao GAO ; Xiaoyan LI ; Xiaoxue WANG ; Huizhu ZHONG ; Qiao WEI ; Yonghua TANG ; Peijian HUANG ; Ruinan SHEN ; Lingyun CHEN ; Jing ZHANG ; Rong FANG ; Wei WEI ; Fengjuan ZHANG ; Gaiyan ZHOU ; Weihong YUAN ; Xi CHEN ; Zhao YANG ; Ying WU ; Wenli XU ; Shuo ZHU ; Liwen ZHANG ; Naying HE ; Weihuan FANG ; Miao ZHANG ; Yu ZHANG ; Huijun JU ; Yaya BAI ; Jun LIU
Chinese Medical Journal 2025;138(22):2907-2916
INTRODUCTION:
Lecanemab has shown promise in treating early Alzheimer's disease (AD), but its safety and efficacy in Chinese populations remain unexplored. This study aimed to evaluate the safety and 6-month clinical outcomes of lecanemab in Chinese patients with mild cognitive impairment (MCI) or mild AD.
METHODS:
In this single-arm, real-world study, participants with MCI due to AD or mild AD received biweekly intravenous lecanemab (10 mg/kg). The study was conducted at Hainan Branch, Ruijin Hospital Shanghai Jiao Tong University School of Medicine. Patient enrollment and baseline assessments commenced in November 2023. Safety assessments included monitoring for amyloid-related imaging abnormalities (ARIA) and other adverse events. Clinical and biomarker changes from baseline to 6 months were evaluated using cognitive scales (mini-mental state examination [MMSE], montreal cognitive assessment [MoCA], clinical dementia rating-sum of boxes [CDR-SB]), plasma biomarker analysis, and advanced neuroimaging.
RESULTS:
A total of 64 patients were enrolled in this ongoing real-world study. Safety analysis revealed predominantly mild adverse events, with infusion-related reactions (20.3%, 13/64) being the most common. Of these, 69.2% (9/13) occurred during the initial infusion and 84.6% (11/13) did not recur. ARIA-H (microhemorrhages/superficial siderosis) and ARIA-E (edema/effusion) were observed in 9.4% (6/64) and 3.1% (2/64) of participants, respectively, with only two symptomatic cases (one ARIA-E presenting with headache and one ARIA-H with visual disturbances). After 6 months of treatment, cognitive scores remained stable compared to baseline (MMSE: 22.33 ± 5.58 vs . 21.27 ± 4.30, P = 0.733; MoCA: 16.38 ± 6.67 vs . 15.90 ± 4.78, P = 0.785; CDR-SB: 2.30 ± 1.65 vs . 3.16 ± 1.72, P = 0.357), while significantly increasing plasma amyloid-β 42 (Aβ42) (+21.42%) and Aβ40 (+23.53%) levels compared to baseline.
CONCLUSIONS:
Lecanemab demonstrated a favorable safety profile in Chinese patients with early AD. Cognitive stability and biomarker changes over 6 months suggest potential efficacy, though high dropout rates and absence of a control group warrant cautious interpretation. These findings provide preliminary real-world evidence for lecanemab's use in China, supporting further investigation in larger controlled studies.
REGISTRATION
ClinicalTrials.gov , NCT07034222.
Humans
;
Alzheimer Disease/drug therapy*
;
Male
;
Female
;
Aged
;
Middle Aged
;
Cognitive Dysfunction/drug therapy*
;
Aged, 80 and over
;
Amyloid beta-Peptides/metabolism*
;
Biomarkers
;
East Asian People
3.Risk of Hospitalization for Genitourinary System Diseases Following Exposure to Cold Spells.
Qing Hua SUN ; Chen CHEN ; Jie BAN ; Han Shuo ZHANG ; Jing Yi SUN ; Hang DU ; Tian Tian LI
Biomedical and Environmental Sciences 2025;38(11):1369-1377
OBJECTIVE:
To assess relationships between cold spells and genitourinary hospitalization risk.
METHODS:
Hospitalization records for genitourinary system diseases (GUDs) from 16 districts in Beijing (2013-2018) were analyzed. Cold spells were defined based on varying intensity thresholds. A two-stage analytical method was employed: first, generalized linear models assessed district-specific associations between cold spells and hospitalizations; second, random-effects meta-analysis aggregated the district-level results. Subgroup analyses were performed by admission type (emergency vs. outpatient), age, and sex.
RESULTS:
A total of 271,579 GUD-related hospitalizations were recorded. Cold spells (p1day2,daily mean temperature below the 1 st percentiles of the daily mean temperature distribution from January 1, 2013, to December 31, 2018, lasting for two or more consecutive days) were linked to a significant rise in hospitalization risks: 1.43 (95% CI: 1.32-1.56) for all GUDs, 1.35 (95% CI: 1.23-1.49) for urinary system diseases, and 1.46 (95% CI: 1.28-1.67) for renal failure, when compared to non-cold spell days. Emergency admissions showed higher risk increases than outpatient admissions.
CONCLUSION
Extreme cold spells significantly elevate hospitalization risks for GUDs. This highlights the urgent need for targeted public health interventions to mitigate cold-related health impacts, especially for vulnerable populations.
Humans
;
Hospitalization/statistics & numerical data*
;
Male
;
Female
;
Cold Temperature/adverse effects*
;
Infant
;
Child, Preschool
;
Middle Aged
;
Adult
;
Child
;
Aged
;
Adolescent
;
Young Adult
;
Beijing/epidemiology*
;
Female Urogenital Diseases/etiology*
;
Male Urogenital Diseases/etiology*
;
Infant, Newborn
;
Risk Factors
4.Effect of over-expression of NR2F2 on biological behaviors of human ovarian cancer SKOV3 cells
Shuo ZHANG ; Yunxiu XIA ; Weiwei CHEN ; Hongliang DONG ; Bingjie CUI ; Cuilan LIU ; Zhiqiang LIU ; Fei WANG ; Jing DU
Journal of Jilin University(Medicine Edition) 2025;51(1):58-67
Objective:To investigate the effect of nuclear receptor subfamily 2 group F member 2(NR2F2)on the biological behaviors of human ovarian cancer SKOV3 cells,and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)Database analyse the expression level of NR2F2 gene in ovarian tissue,and analyse its correlation with clinical prognosis of ovarian cancer patients.The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression(NR2F2 OE)group,which were transfected with mCherry control virus and NR2F2 OE over-expression virus,respectively,when the cell deusity reached 70%,and the stable transfection SKOV3 cell lines were screened with puromycin(puro)48h lafter.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the transfection efficiencies of the cells;RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2(SOX2)mRNA in the cells in two groups;Western blotting method was used to detect the expression levels of NR2F2,ATP-binding cassette superfamily G member 2(ABCG2),and programmed cell death 1-ligand 1(PD-L1)protcins in the cells in two groups.CCK-8 assay was used to detect the proliferation activities of the cells in two groups;Wound assay was used to detect the migration rates of the cells in two groups;Transwell chamber assay was used to detect the number of transmembrane cells;Spheroidization assay was used to detect the numbers of spheroids in the cells;peripheral blood mononuclear cells(PBMCs)-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells;CCK-8 assay was used to detect the half maximal inhibitory concentration(IC50)of paclitaxel(PTX)and carboplatin(CBP).Results:Compared with normal ovarian tissue,the expression level of NR2F2 gene in ovarian tumor tissue was decreased(P<0.05),and decreased with the improvement of clinical pathological grading of ovarian tumor.The patients with higher expression level of NR2F2 gene had better clincal prognosis.The SKOV3 cells with NR2F2 over-expresson were successfully constructed,and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group(P<0.001).The CCK-8 assay results showed that compared with control group,the proliferation activities of the cells in NR2F2 OE group were decreased at different time points(1,2,3,and 4 d)(P<0.05 or P<0.01).The cell wound assay results showed that compared with control group,the migration rate of the cells in NR2F2 OE group was decreased(P<0.001).The Transwell assay results showed that compared with control group,the number of transmembrane cells in NR2F2 OE group was decreased(P<0.01).Compared with control group,the number of the spheroids in NR2F2 OE group was decreased(P<0.05),and the expression levels of SOX2 mRNA(P<0.01)and protein(P<0.001)were increased.Compared with control group,the relative density of surviving tumor cells in NR2F2 OE group was decreased,but the difference was not significant(P<0.05),and the expression level of PD-L1 protein was decreased(P<0.05).Compared with control group,the proliferation activities of cells in NR2F2 OE group were decreased(P<0.05),and the drug sensitivities of the cells to PTX and CBP were enhanced(P<0.05);the IC50 of PTX was significantly reduced,while the IC50of CBP could not be calculated due to excessively high drug concentration;the expression level of ABCG2 protein was decreased(P<0.05).Conclusion:The over-expression of NR2F2 may inhibit the proliferation,migration,and invasion of the human ovarian cancer SKOV3 cells,decrease the expression levels of SOX2,PD-L1 and ABCG2 proteins,suppress the stemness and immune evasion ability of the SKOV3 cells,and enhance the sensitivities of the SKOV3 cells to PTX and CBP.
5.Long non-coding RNA LINC00973 promotes migration,invasion and distal metastasis of epithelial ovarian cancer and its molecular mechanism
Yunxiu XIA ; Shuo ZHANG ; Huanhai ZHANG ; Fei WANG ; Hongliang DONG ; Jing DU
Journal of Jilin University(Medicine Edition) 2025;51(4):866-878
Objective:To discuss the effect of long non-coding RNA(lncRNA)LINC00973 on the migration,invasion,and distant metastasis of epithelial ovarian cancer,and to clarify its molecular mechanism.Methods:The human ovarian cancer SKOV3 and OVCAR3 cells were divided into EF1a-FH empty vector control group(control group),LINC00973 overexpression group(LINC00973 OE group),U6-shRNA empty vector control group(SHV group),and LINC00973 knockdown group(LINC00973 KD group),and were transfected with lentivirus containing nonsense sequence(pLent-EF1a-FH-CMV-copGFP-P2A-Puro),LINC00973 overexpression,nonsense sequence(pLent-U6-shRNA-CMV-copGFP-P2A-Puro)and LINC00973 shRNA,respectively,followed by puromycin screening to obtain stably transfected SKOV3 and OVCAR3 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of target genes in the cells in various groups;wound healing assay was used to detect the migration rate of the cells in various groups;Transwell chamber assay was used to detect the number of transmembrane cells in various groups;The mice were divided into control group(WT group),LINC00973 OE group,and LINC00973 KD group,with 4 mice in each group.SKOV3 wild-type cells,LINC00973 OE cells,and LINC00973 KD cells were intraperitoneally injected into the mice in various groups,respectively,to establish the epithelial ovarian cancer intraperitoneal implantation and metastasis model;HE staining was used to observe the morphology of the colon and liver tissues of the mice in various groups;RNA-secquencing(RNA-seq)was used to analyze the differentially expressed genes between SHV and LINC00973 KD groups in the SKOV3 cell line;RT-qPCR method was used to detect the mRNA expression levels of LINC00973 in the normal ovarian epithelial cells IOSE-80 and epithelial ovarian cancer cells SKOV3,A2780 and OVCAR3,the mRNA expression levels of LINC00973,Vimentin,Snail family transcriptional repressor 1(Snail),Twist family basic helix-loop-helix transcription factor 1(Twist),zinc finger E-box binding homeobox 1(ZEB1),zinc finger E-box binding homeobox 2(ZEB2),CXCL8 and matrix metalloproteinase(MMP)16 in the cells in various groups,and the mRNA expression levels of LINC00973,Vimentin and Twist in liver and colon tissues of the mice in various groups.Results:Compared with normal ovarian epithelial cells IOSE-80,the expression level of LINC00973 mRNA in the epithelial ovarian cancer cells SKOV3,OVCAR3 and A2780 was significantly increased(P<0.01),with the highest expression level of LINC00973 in SKOV3 and OVCAR3 cells,which were therefore selected for subsequent experiments.In SKOV3 and OVCAR3 cells,compared with control group,the expression level of LINC00973 mRNA in the cells in LINC00973 OE group was increased(P<0.01);compared with SHV group,the expression level of LINC00973 mRNA in the cells in LINC00973 KD group was decreased(P<0.05 or P<0.01),indicating successful construction of LINC00973 overexpression and knockdown cell lines.In SKOV3 cells,compared with control group,the mRNA expression levels of Vimentin and Twist in LINC00973 OE group were increased(P<0.05 or P<0.01),while no significant difference was observed in Snail mRNA expression level(P>0.05);compared with SHV group,the mRNA expression levels of Vimentin,Snail and Twist in LINC00973 KD group were decreased(P<0.01).In OVCAR3 cells,compared with control group,the mRNA expression levels of Vimentin,Snail and Twist in LINC00973 OE group were increased(P<0.01);compared with SHV group,the expression levels of Vimentin,Snail,and mRNA Twist in LINC00973 KD group were decreased(P<0.01).The wound healing assay results showed that compared with control group,the wound healing rates of the SKOV3 and OVCAR3 cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the wound healing rates of the cells in LINC00973 KD group were significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with control group,the numbers of transmembrane cells of the SKOV3 and OVCAR3 cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the numbers of transmembrane cells in LINC00973 KD group were significantly decreased(P<0.01).Compared with WT group,the number of peritoneal nodules in LINC00973 OE group was increased,with rough liver surface and multiple nodules formed on mesentery and colon surface,and the expression levels of LINC00973,Vimentin,and Twist mRNA in colon tissue were increased(P<0.01);compared with WT group,no nodules were formed in the peritoneal cavity of LINC00973 KD group,with smooth liver surface,no nodules in liver tissue,and decreased expression levels of LINC00973,Vimentin,and Twist mRNA,and no nodules were observed on mesentery and colon surface.The HE staining results showed that compared with WT group,the multiple lesions were observed in liver and colon tissues in LINC00973 OE group,manifested as uneven cell size,irregular shape,unclear cell boundaries,increased nuclear division,and uneven red staining in cytoplasm,while in LINC00973 KD group,the cells in liver and colon tissues were arranged neatly with regular shape,and uniform distribution of nuclei and cytoplasm.The RNA-seq results showed that compared with SHV group,no key signaling pathways related to tumor metastasis were enriched in LINC00973 KD group,and the transcription levels of metastasis-related genes CXCL8,MMP16,ZEB1,and ZEB2 were decreased.The RT-qPCR results showed that compared with control group,the expression levels of ZEB1,ZEB2,CXCL8,and MMP16 mRNA in the cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the expression levels of ZEB1,ZEB2,CXCL8,and MMP16 mRNA in the cells in LINC00973 KD group were significantly decreased(P<0.01).Conclusion:LINC00973 can up-regulate the expression of metastasis-related factors Vimentin,Snail,Twist,ZEB1,ZEB2,CXCL8,and MMP16,and promote the migration,invasion,and distant metastasis of epithelial ovarian cancer.
6.Efficacy and safety of recombinant human anti-SARS-CoV-2 monoclonal antibody injection(F61 injection)in the treatment of patients with COVID-19 combined with renal damage:a randomized controlled exploratory clinical study
Ding-Hua CHEN ; Chao-Fan LI ; Yue NIU ; Li ZHANG ; Yong WANG ; Zhe FENG ; Han-Yu ZHU ; Jian-Hui ZHOU ; Zhe-Yi DONG ; Shu-Wei DUAN ; Hong WANG ; Meng-Jie HUANG ; Yuan-Da WANG ; Shuo-Yuan CONG ; Sai PAN ; Jing ZHOU ; Xue-Feng SUN ; Guang-Yan CAI ; Ping LI ; Xiang-Mei CHEN
Chinese Journal of Infection Control 2024;23(3):257-264
Objective To explore the efficacy and safety of recombinant human anti-severe acute respiratory syn-drome coronavirus 2(anti-SARS-CoV-2)monoclonal antibody injection(F61 injection)in the treatment of patients with coronavirus disease 2019(COVID-19)combined with renal damage.Methods Patients with COVID-19 and renal damage who visited the PLA General Hospital from January to February 2023 were selected.Subjects were randomly divided into two groups.Control group was treated with conventional anti-COVID-19 therapy,while trial group was treated with conventional anti-COVID-19 therapy combined with F61 injection.A 15-day follow-up was conducted after drug administration.Clinical symptoms,laboratory tests,electrocardiogram,and chest CT of pa-tients were performed to analyze the efficacy and safety of F61 injection.Results Twelve subjects(7 in trial group and 5 in control group)were included in study.Neither group had any clinical progression or death cases.The ave-rage time for negative conversion of nucleic acid of SARS-CoV-2 in control group and trial group were 3.2 days and 1.57 days(P=0.046),respectively.The scores of COVID-19 related target symptom in the trial group on the 3rd and 5th day after medication were both lower than those of the control group(both P<0.05).According to the clinical staging and World Health Organization 10-point graded disease progression scale,both groups of subjects improved but didn't show statistical differences(P>0.05).For safety,trial group didn't present any infusion-re-lated adverse event.Subjects in both groups demonstrated varying degrees of elevated blood glucose,elevated urine glucose,elevated urobilinogen,positive urine casts,and cardiac arrhythmia,but the differences were not statistica-lly significant(all P>0.05).Conclusion F61 injection has initially demonstrated safety and clinical benefit in trea-ting patients with COVID-19 combined with renal damage.As the domestically produced drug,it has good clinical accessibility and may provide more options for clinical practice.
7.Implementation of surveillance,prevention and control of healthcare-asso-ciated infection in maternal and child healthcare institutions:A nation-wide investigation report
Shuo LI ; Xi YAO ; Hui-Xue JIA ; Wei-Guang LI ; Xun HUANG ; Shu-Mei SUN ; Xi CHENG ; Qing-Lan MENG ; Xiang ZHANG ; Jing-Ping ZHANG ; Ya-Wei XING ; Qing-Qing JIANG ; Lian-Xuan WU ; Bing-Li ZHANG ; Xiao-Jing LIU ; Liu-Yi LI
Chinese Journal of Infection Control 2024;23(3):323-329
Objective To investigate the implementation of surveillance,prevention and control measures for healthcare-associated infection(HAI)in maternal and child healthcare(MCH)institutions,and provide policy evi-dence for optimizing HAI prevention and control in MCH institutions.Methods Stratified sampling was conducted among the MCH institutions at provincial,municipal and county levels in 8 provinces/autonomous regions.A uni-fied questionnaire was designed and the online survey was conducted through"Questionnaire Star".Results The data from 123 MCH institutions were included in the analysis.90.24%of the MCH institutions carried out compre-hensive surveillance on HAI.The ratios of MCH institutions which implemented targeted surveillance on HAI in neonatal intensive care unit(NICU),surgical site infection,multidrug-resistant organisms(MDROs)and HAI in intensive care units(non-NICU excluded)were 89.66%,85.96%,80.77%,and 74.19%,respectively.51.22%MCH institutions adopted information surveillance system on HAI cases.94.31%MCH institutions carried out surveillance on hand hygiene compliance.Over 90%MCH institutions carried out surveillance on environment hy-giene in high-risk departments.71.54%MCH institutions conducted centralized cleaning,disinfection,sterilization and supply for reusable medical instruments in the central sterile supply department(CSSD).Over 90%MCH insti-tutions established three-level pre-examination triage systems.86.18%set up transitional wards.MCH institutions generally adopted a management model with established effective communication,full appointment visits,and sepa-rate visits for special medical groups,such as registered pregnant women,high-risk newborns,healthcare groups,and long-term rehabilitation patients.However,the ratio of institutions conducting on-line follow-up visits was less than 50%.Conclusion MCH institutions have generally carried out comprehensive and targeted surveillance on HAI.Information surveillance need to be facilitated.Hand hygiene and environmental hygiene surveillance has been popularized to a certain extent at all levels of MCH institutions.The cleaning,disinfection,sterilization,and supply processes of reusable medical devices in a few MCH institutions are not standardized.Special medical populations get effective management.On-line healthcare is to be further promoted.
8.Clinical and immunological characteristics of acute viral infection-related encephalopathy
Jianzhao ZHANG ; Caihui MA ; Jing SUN ; Dongqing LI ; Zhao LIU ; Shuo MIAO ; Hui JIAO ; Jian YANG
Chinese Journal of Applied Clinical Pediatrics 2024;39(4):298-302
Objective:To analyze the clinical and immunological characteristics of children with acute viral infection-related encephalopathy.Methods:Case-control study.A retrospective analysis was conducted on the clinical data of children diagnosed with acute viral infection-related encephalopathy during hospitalization at the Children′s Hospital, Capital Institute of Pediatrics from January 2020 to January 2023.According to the last follow-up modified Rankin scale (mRS) score, these children were divided into a good prognosis group (mRS score ≤2) and a poor prognosis group (mRS score >2), and the clinical and immunological characteristics of the children with different prognoses were analyzed.The binary Logistic regression was used to analyze the risk factors for poor prognosis.Results:A total of 28 children with acute viral infection-related encephalopathy aged 4 months to 11 years were included.There were 16 males (57%) and 12 females (43%). Among the preinfection viruses, there were 16 children of Corona virus disease 2019, 8 children of influenza A virus, 3 children of influenza B virus, and 1 child of norovirus.Among them, there were 21 children with acute necrotizing encephalopathy, 4 children with acute encephalopathy with biphasic seizures and late reduced diffusion, 2 children with mild encephalitis with a reversible splenial lesion, and 1 child with hemorrhagic shock and encephalopathy syndrome.Among the first symptoms, 24 children (85.7%) had consciousness disorders, 23 children (82.1%) had seizures, 17 children (60.7%) had speech disorders, 11 children (39.3%) had involuntary movements, and 10 children (35.7%) had abnormal mental behavior.For the site of lesion, the cranial nuclear magnetic resonance imaging revealed 17 in the thalamus, 10 in the brainstem, 9 in the basal ganglia, 8 in the cerebellar hemisphere, and 4 in the corpus callosum.In the last follow-up evaluation, 17 children had a mRS score of >2, and 11 children had a mRS score of ≤2.Univariate analysis showed that disturbance of consciousness, seizure cluster, brain stem lesion, absolute value of serum T lymphocytes, cerebrospinal fluid(CSF) protein, CSF cytokines [interleukin(IL)-1β, IL-6 and IL-8]were higher in the poor prognosis group than those in the good prognosis group.Multivariate Logistic regression analysis indicated that brain stem disease, CSF IL-1β and T lymphocyte absolute number were independent risk factors for poor prognosis.Conclusions:Brain stem lesions, cerebrospinal fluid IL-1β and the absolute number of T lymphocytes have predictive value for the prognosis of acute viral infection-associated encephalopathy.The more severe the conditions, the lower the T lymphocytes, and the higher the cytokines in some cerebrospinal fluid.
9.PMA-ddPCR method for detecting high ethanol-producing klebsiella pneumoniae in viable but non-culturable state
Shuo ZHAO ; Chenpu DOU ; Jian ZHANG ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(7):998-1003
Objective:To establish an absolute quantitative method for high ethanol-producing klebsiella pneumoniae in a viable non-culturable (VBNC) state. Methods:High ethanol-producing Klebsiella pneumonia was induced to enter the VBNC state and then the ethanol production was evaluated. A PMA-ddPCR method was established to count the copies of live cell genes in the VBNC state of high ethanol-producing Klebsiella pneumoniae using single-copy genes. Further, the sensitivity and adaptability of ddPCR for detecting low-concentration samples were evaluated in VBNC fecal simulation. Results:The lower detection limit of ddPCR for quantitative analysis of high ethanol-producing Klebsiella pneumoniae gradient diluent was 10 times that of qPCR. At low temperature and low nutritional state, high ethanol-producing Klebsiella pneumoniae entered the VBNC state on the 45 th day. The quantitative results of PMA-ddPCR on VBNC state cells were (5.46±0.05) log 10 DNA copies/ml. The ethanol production in the VBNC state was<2.2 mmol/L and the ability to produce ethanol was restored after recovery. The minimum detection limit for ddPCR in fecal simulated samples with VBNC state was 3.2 log 10 DNA copies/ml. Conclusion:The ddPCR detection method for high ethanol-producing Klebsiella pneumoniae with VBNC state has good sensitivity and adaptability, and can be used for the detection of VBNC state cells in clinical samples.
10.PMA-ddPCR method for detecting high ethanol-producing klebsiella pneumoniae in viable but non-culturable state
Shuo ZHAO ; Chenpu DOU ; Jian ZHANG ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(7):998-1003
Objective:To establish an absolute quantitative method for high ethanol-producing klebsiella pneumoniae in a viable non-culturable (VBNC) state. Methods:High ethanol-producing Klebsiella pneumonia was induced to enter the VBNC state and then the ethanol production was evaluated. A PMA-ddPCR method was established to count the copies of live cell genes in the VBNC state of high ethanol-producing Klebsiella pneumoniae using single-copy genes. Further, the sensitivity and adaptability of ddPCR for detecting low-concentration samples were evaluated in VBNC fecal simulation. Results:The lower detection limit of ddPCR for quantitative analysis of high ethanol-producing Klebsiella pneumoniae gradient diluent was 10 times that of qPCR. At low temperature and low nutritional state, high ethanol-producing Klebsiella pneumoniae entered the VBNC state on the 45 th day. The quantitative results of PMA-ddPCR on VBNC state cells were (5.46±0.05) log 10 DNA copies/ml. The ethanol production in the VBNC state was<2.2 mmol/L and the ability to produce ethanol was restored after recovery. The minimum detection limit for ddPCR in fecal simulated samples with VBNC state was 3.2 log 10 DNA copies/ml. Conclusion:The ddPCR detection method for high ethanol-producing Klebsiella pneumoniae with VBNC state has good sensitivity and adaptability, and can be used for the detection of VBNC state cells in clinical samples.

Result Analysis
Print
Save
E-mail