Objective　To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.

To clarify the species, pathogenicity, and distribution of the pathogens causing the root rot of Atractylodes lancea in Hubei province, the tissue separation method was used to isolate the pathogens from root rot samples in the main planting areas of A. lancea in Hubei. Based on the preliminary identification of the Fusarium genus by the internal transcribed spacer(ITS) sequence, three housekeeping genes, EF1/EF2, Btu-F-FO1/Btu-F-RO1, and FF1/FR1, were amplified and sequenced. Subsequently, a phylogenetic tree was constructed based on these TEF gene sequences to classify the pathogens. The pathogenicity of these strains was determined using the root irrigation method. A total of 194 pathogen strains were isolated using the tissue separation method. Molecular identification using the three housekeeping genes identified the pathogens as F. solani, F. oxysporum, F. commune, F. equiseti, F. tricinctum, F. redolens, F. fujikuroi, F. avenaceum, F. acuminatum, and F. incarnatum. Among them, F. solani and F. oxysporum were the dominant strains, widely distributed in multiple regions, with F. solani accounting for approximately 54% of the total isolated strains and F. oxysporum accounting for approximately 34%. Other strains accounted for a relatively small proportion, totaling approximately 12%. The results of pathogenicity determination showed that there were certain differences in pathogenicity among strains. The analysis of the pathogenicity differentiation of the widely distributed F. solani and F. oxysporum strains revealed that these dominant strains in Hubei were mainly highly pathogenic. This study determined the species, pathogenicity, and distribution of the pathogens causing the root rot of A. lancea in Hubei province. The results provide a scientific basis for further understanding the root rot of A. lancea and its epidemic occurrence and scientifically preventing and controlling this disease.
Plant Diseases/microbiology* ; Atractylodes/microbiology* ; Phylogeny ; Plant Roots/microbiology* ; Fusarium/classification* ; China ; Virulence ; Fungal Proteins/genetics*

OBJECTIVE: The present study evaluated the effects of deep acupuncture at Weizhong acupoint (BL40) on bladder function and brain activity in a rat model of overactive bladder (OAB), and investigated the possible mechanisms around the acupuncture area that initiate the effects of acupuncture. METHODS: Adult female Sprague-Dawley rats were randomly divided into six groups, comprising a control group, model group, group treated with deep acupuncture at BL40, group treated with shallow acupuncture at BL40, group treated with acupuncture at non-acupoint next to BL40, and group treated with acupuncture at Xuanzhong (GB39). Urodynamic evaluation was used to observe the urination, and functional magnetic resonance imaging was used to observe the brain activation. The mechanism of acupuncture at BL40 in regulating bladder function was explored by toluidine blue staining and enzyme-linked immunosorbent assay, and the mechanism was verified by stabilizing mast cells (MCs) or blocking tibial nerve. RESULTS: Deep acupuncture at BL40 significantly increased the intercontraction interval in OAB rats and enhanced the mean amplitude of low frequency fluctuation of primary motor cortex (M1), periaquaductal gray matter (PAG), and pontine micturition center (PMC). It also increased the zero-lag functional connectivity between M1 and PAG and between PAG and PMC. Shallow acupuncture at BL40 and acupuncture at non-acupoint or GB39 had no effect on these indexes. Further studies suggested that deep acupuncture at BL40 increased the number and degranulation rate of MCs as well as the contents of 5-hydroxytryptamine, substance P, and histamine in the tissues around BL40. Blocking the tibial nerve by lidocaine injection or inhibiting MC degranulation by sodium cromoglycate injection obstructed the effects of acupuncture on restoring urinary function and modulating brain activation in OAB rats. CONCLUSION Deep acupuncture at BL40 may be more effective for inhibiting OAB by promoting degranulation of MCs around the acupoint and stimulating tibial nerve, thereby regulating the activation of the brain area that controls the lower urinary tract. Please cite this article as: Liu X, Zhang CY, Du XY, Li SS, Wang YQ, Zheng Y, Deng HZ, Fang XQ, Li JY, Wang ZQ, Xu SF, Mi YQ. Acupuncture at Weizhong (BL40) attenuates acetic acid-induced overactive bladder in rats by regulating brain neural activity through the modulation of mast cells and tibial nerves. J Integr Med. 2025; 23(1): 46-55.
Animals ; Urinary Bladder, Overactive/physiopathology* ; Mast Cells/physiology* ; Rats, Sprague-Dawley ; Female ; Acupuncture Therapy ; Acupuncture Points ; Rats ; Brain/physiopathology* ; Tibial Nerve/physiopathology* ; Acetic Acid ; Urinary Bladder/physiopathology*

OBJECTIVES: To investigate the prognostic value of molecular minimal residual disease (Mol-MRD) monitored before and after allogeneic hematopoietic stem cell transplantation (HSCT) in pediatric acute myeloid leukemia (AML). METHODS: Clinical data of 71 pediatric AML patients who underwent HSCT between August 2016 and December 2023 were analyzed. Mol-MRD levels were dynamically monitored in MRD-positive patients, and survival outcomes were evaluated. RESULTS: No significant difference in the 3-year overall survival (OS) rate was observed between patients with pre-HSCT Mol-MRD ≥0.01% and <0.01% (77.3% ± 8.9% vs 80.4% ± 7.9%, P=0.705). However, patients with pre-HSCT Mol-MRD <1.75% had a significantly higher 3-year OS rate than those with Mol-MRD ≥1.75% (86.6% ± 5.6% vs 44.4% ± 16.6%, P=0.020). The median Mol-MRD level in long-term survivors was significantly lower than in non-survivors [0.61% (range: 0.04%-51.58%)] vs 10.60% (range: 1.90%-19.75%), P=0.035]. Concurrent flow cytometry-based MRD positivity was significantly higher in non-survivors (80% vs 24%, P=0.039). There was no significant difference in the 3-year overall survival rate between patients with Mol-MRD ≥0.01% and those with <0.01% at 30 days post-HSCT (P=0.527). For children with Mol-MRD <0.22% at 30 days post-HSCT, the 3-year overall survival rate was 80.4% ± 5.9%, showing no significant difference compared to those with molecular negativity (87.0% ± 7.0%) (P=0.523). CONCLUSIONS Patients with pre-HSCT Mol-MRD <1.75% or post-HSCT Mol-MRD <0.22% may achieve long-term survival outcomes comparable to Mol-MRD-negative cases through HSCT and targeted interventions.
Humans ; Hematopoietic Stem Cell Transplantation ; Neoplasm, Residual ; Leukemia, Myeloid, Acute/genetics* ; Child ; Male ; Female ; Child, Preschool ; Prognosis ; Adolescent ; Infant ; Transplantation, Homologous

Objective To observe the effects of Yuebi Decoction on adriamycin-induced nephrotic syndrome(NS)and adhesion molecule levels in rats;To explore the possible mechanism of its inhibition of NS edema.Methods A NS rat model was established by a one-time tail vein injection of 6.5 mg/kg adriamycin.The rats were randomly divided into normal group,model group,prednisone group and Yuebi Decoction group.The prednisone group and Yuebi Decoction group were given 5 mg/kg prednisone and 22.2 g/kg Yuebi Decoction by gavage,respectively.The normal group and the model group were given an equal amount of normal saline by gavage,once a day for 2 consecutive weeks.The wet and dry weight ratios of skin and kidney were detected,24 hour urine total protein quantification(24 hUTP)and the contents of blood urea nitrogen(BUN),serum creatinine(SCr),total cholesterol(TC),albumin(ALB),hemoglobin(HB)were detected,HE and PAS staining was used to observe the morphology of renal tissue,ELISA was used to detect vascular cell adhesion molecule-1(VCAM-1)and intercellular adhesion molecule-1(ICAM-1)contents in renal tissue,the positive expressions of VCAM-1 and ICAM-1 were detected by immunohistochemistry,the expressions of VCAM-1,Rac1,signal transducer and activator of transcription 3(STAT3)and p-STAT3 proteins were detected by Western blot.Results Compared with the normal group,the wet dry weight ratio of the skin and kidney of the model group rats significantly increased(P<0.01),and the 24 hUTP,SCr,BUN and TC contents significantly increased(P<0.01),while the ALB and HB contents significantly decreased(P<0.01);with glomerular balloon atrophy,the renal tubules exhibit protein casts,inflammatory cell infiltration in renal interstitium,and significant increase in the contents of VCAM-1,ICAM-1 and protein expressions of VCAM-1,Rac1,p-STAT3 in renal tissue(P<0.01).Compared with the model group,the wet dry weight ratio of the skin and kidneys of rats in the prednisone group and Yuebi Decoction group was significantly reduced(P<0.05,P<0.01),and the 24 hUTP,SCr,BUN and TC were significantly reduced(P<0.01),while the contents of ALB and HB significantly increased(P<0.01);the infiltration of inflammatory cells in renal tissue was reduced,the protein casts decreased,and the contents of VCAM-1 and ICAM-1 in renal tissue,as well as the expressions of VCAM-1,Rac1 and p-STAT3 proteins significantly decreased(P<0.05,P<0.01).Conclusion Yuebi Decoction can alleviate edema,reduce urine protein,and improve capillary permeability in NS rats.Its mechanism is related to inhibiting the Rac1/STAT3 signaling pathway and reducing the expression of renal tissue adhesion molecules.

Objective To investigate the size variation in the first metatarsal of extant primates.Methods In this study,we analyzed 135 first metatarsal specimens across 12 primate genera,quantifying eight linear measurements,articular surface areas,mid-shaft cross-sectional area,total surface area,volume,and derived indices.Multivariate patterns were assessed through mean-based correspondence analysis and principal component analysis(PCA).Results Eulemur,Otolemur,Cebus,and Perodicticus exhibited a relatively high metatarsal surface-area-to-volume ratio.Perodicticus additionally showed a low articular facet index.Propithecus,Colobus,and Macaca displayed lower values for metatarsal shaft robusticity,the ratio of shaft cross-sectional area to base articular surface area,and the proximal articular facet index.Nasalis possessed a relatively high articular facet index.Pongo,Pan,Gorilla,and Homo sapiens were characterized by higher metatarsal shaft robusticity and a lower metatarsal surface-area-to-volume ratio.Principal component analysis revealed that the 12 extant primate genera could be broadly divided into two groups.Group 1 comprised Pongo,Pan,Gorilla and H.sapiens,although H.sapiens formed a distinct cluster relative to the extant great apes.Group 2 included Eulemur,Otolemur,Perodicticus,Propithecus,Cebus,Colobus,Macaca and Nasalis.Conclusion The relative sizedistribution of the first metatarsal provides some reference value for classifying extant primates.However,it demonstrates no clear correlations with specific locomotor patterns or foot grasping ability.

Objective:To explore the trajectory of depressive symptoms and its related factors in rural women in Yunnan from first trimester of pregnancy to 42 days after childbirth.Methods:The longitudinal study design was used,and 213 pregnant women from a rural county of Yunnan were selected.The Edinburgh Postnatal Depression Scale(EPDS),Generalized Anxiety Disorder(GAD-7),and Perceived Social Support Scale(PSSS)were used to surveyed in their first trimester of pregnancy,and followed up in their second and third trimesters of pregnancy and 42 days after childbirth.The analysis was conducted using latent growth curve models,latent class growth models,and multivariate logistic regression models.Results:The results of the latent growth curve models showed a gradual decrease in perinatal depressive symptom scores over time.The latent class growth models revealed three distinct la-tent classes:persistent high depression score group(1.41％),stable low depression score group(91.08％),and fluctuating depression symptoms group(7.51％).Multivariate logistic regression indicated that women with lower education levels(OR=4.04),anxiety symptoms(OR=4.12),and lower social support(OR=3.12)were more likely to be classified into the persistent high depression score group and fluctuating depression symptoms group.Conclusions:The trajectories of perinatal depression symptoms among rural women in Yunnan could be di-vided into three heterogeneous subgroups.Women with low education levels,anxiety symptoms,and lack of social support are more likely to be detected with perinatal depression symptoms.

Objective To observe the effects of Yuebi Decoction on adriamycin-induced nephrotic syndrome(NS)and adhesion molecule levels in rats;To explore the possible mechanism of its inhibition of NS edema.Methods A NS rat model was established by a one-time tail vein injection of 6.5 mg/kg adriamycin.The rats were randomly divided into normal group,model group,prednisone group and Yuebi Decoction group.The prednisone group and Yuebi Decoction group were given 5 mg/kg prednisone and 22.2 g/kg Yuebi Decoction by gavage,respectively.The normal group and the model group were given an equal amount of normal saline by gavage,once a day for 2 consecutive weeks.The wet and dry weight ratios of skin and kidney were detected,24 hour urine total protein quantification(24 hUTP)and the contents of blood urea nitrogen(BUN),serum creatinine(SCr),total cholesterol(TC),albumin(ALB),hemoglobin(HB)were detected,HE and PAS staining was used to observe the morphology of renal tissue,ELISA was used to detect vascular cell adhesion molecule-1(VCAM-1)and intercellular adhesion molecule-1(ICAM-1)contents in renal tissue,the positive expressions of VCAM-1 and ICAM-1 were detected by immunohistochemistry,the expressions of VCAM-1,Rac1,signal transducer and activator of transcription 3(STAT3)and p-STAT3 proteins were detected by Western blot.Results Compared with the normal group,the wet dry weight ratio of the skin and kidney of the model group rats significantly increased(P<0.01),and the 24 hUTP,SCr,BUN and TC contents significantly increased(P<0.01),while the ALB and HB contents significantly decreased(P<0.01);with glomerular balloon atrophy,the renal tubules exhibit protein casts,inflammatory cell infiltration in renal interstitium,and significant increase in the contents of VCAM-1,ICAM-1 and protein expressions of VCAM-1,Rac1,p-STAT3 in renal tissue(P<0.01).Compared with the model group,the wet dry weight ratio of the skin and kidneys of rats in the prednisone group and Yuebi Decoction group was significantly reduced(P<0.05,P<0.01),and the 24 hUTP,SCr,BUN and TC were significantly reduced(P<0.01),while the contents of ALB and HB significantly increased(P<0.01);the infiltration of inflammatory cells in renal tissue was reduced,the protein casts decreased,and the contents of VCAM-1 and ICAM-1 in renal tissue,as well as the expressions of VCAM-1,Rac1 and p-STAT3 proteins significantly decreased(P<0.05,P<0.01).Conclusion Yuebi Decoction can alleviate edema,reduce urine protein,and improve capillary permeability in NS rats.Its mechanism is related to inhibiting the Rac1/STAT3 signaling pathway and reducing the expression of renal tissue adhesion molecules.

Objective To observe the protective effect of Yuebi Decoction on adriamycin(ADR)-induced mouse glomerular endothelial cells(MGEC),and to explore the possible mechanism of action.Methods MGEC were stimulated with ADR at a concentration of 1.25 μmol·L-1 for 12 h.MGEC were divided into five groups:normal group,model group,and Yuebi Decoction low,medium,and high dosage groups(5%,10%,and 20%of drug-containing serum);the viability of cells was detected by CCK-8 assay,the cell mobility was detected by Transwell assay.The relative expression levels of MGEC VCAM-1,Rac1,STAT3,and p-STAT3 were detected by Western blot.The expression of endothelial marker CD31,VCAM-1 and ICAM-1 was detected by immunofluorescence.Results Compared with the normal group,the endothelial cell migration rate was decreased in the model group(P<0.01),the protein expression levels of VCAM-1,ICAM-1,Rac1 and p-STAT 3 were significantly increased(P<0.05,P<0.01).The endothelial cell migration rate of the high-dose group of Yuebi Decoction was significantly increased compared with that of the model group(P<0.05).VCAM-1,ICAM-1,Rac1 and p-STAT3 protein expression levels of the drug-containing serum of Yuebi Decoction each group were significantly reduced(P<0.05,P<0.01).Conclusion Yuebi Decoction inhibited ADR-induced adhesion molecule expression in MGEC,alleviated endothelial injury,and improved endothelial cell migration rate.Its mechanism may be related to the inhibition of the Rac1/STAT3 signaling pathway.

[Purpose]To analyze the compliance and its influencing factors of lung cancer screening using low-dose computed tomography(LDCT)among high-risk population in urban districts of Chongqing from 2013 to 2021.[Methods]The lung cancer screeing of Cancer Early Diagnosis and Treatment Project was conducted among permanent residents aged 40～69 years old from 14 urban districts of Chongqing selected by cluster sampling method from 2013 to 2021.The questionnaire survey was performed to assess the risk level of lung cancer,and individuals with high risk were advised to have LDCT examination.The compliance rate of LDCT examination among high-risk populations was calculated and compared using Chi-square test among residents with different de-mographic features;the influencing factors of compliance was analyzed with generalized linear mixed models.[Results]A total of 316 066 residents completed the risk assessment questionnaire survey,52 858 people were assessed as high-risk(17.17％).Among the high-risk population,20 398 completed LDCT screening,with an overall compliance rate of 38.59％.The generalized linear mixed model showed that male participants(OR=0.871,95％CI:0.823～0.922)and smokers(light smokers:OR=0.829,95％CI:0.775～0.886;heavy smokers:OR=0.842,95％CI:0.792～0.896)had lower compliance rates;while people with higher education level(OR=1.347,95％CI:1.265～1.435),occupational exposure to harmful substances(OR=1.400,95％CI:1.340～1.463),passive smoking for 20 years or more(OR=1.472,95％CI:1.376～1.576),infrequent physical exercise(OR=1.203,95％CI:1.152～1.256),family history of lung cancer(OR=2.312,95％CI:2.201～2.429),and those having media promotion by community staff(OR=1.365,95％CI:1.223～1.524),and trained community staff(OR=1.343,95％CI:1.227～1.470)had higher compliance rates.Comorbidities were also factors influencing compliance,and there was an increasing trend of compliance rate with the increase of comorbidity numbers(P＜0.001).[Conclusion]The compli-ance rate of LDCT examination for lung cancer screening in Chongqing needs to be improved,and more precise health education should be implemented for groups with different characteristics to improve the compliance among high-risk population.