Humans ; Gastrointestinal Microbiome ; Constipation/therapy* ; Probiotics/therapeutic use* ; Patients

AIM: To compare the efficacy of intravitreal injection of ranibizumab(IVR)and intravitreal injection of conbercept(IVC)in children with retinopathy of prematurity(ROP).METHODS: Retrospective study. A total of 1 100 eyes with ROP treated with intravitreal anti-VEGF at our hospital from January 2015 to June 2023 were included. According to the different therapeutic drugs, the children were divided into two groups: IVR group and IVC group. According to the degree of ROP, the patients were divided into three groups: aggressive ROP(A-ROP), Zone Ⅰ type 1 ROP and Zone Ⅱ type 1 ROP. The reactivation and retreatment between the two groups were compared after propensity score matching(PSM)analysis, and they were followed-up for at least 3 mo after surgery.RESULTS: In Zone Ⅱ type 1 ROP, there was a statistically significant difference in the rates of reactivation and retreatment between the IVR and IVC groups(P<0.05); however, in A-ROP and Zone I type 1 ROP, there were no statistically significant differences in the rates of reactivation and retreatment between the two groups(P>0.05). The risk of reactivation and retreatment of Zone I type 1 ROP was higher than the Zone II type 1 ROP. Furthermore, the use of drugs and corrected gestational age of first treatment were influencing factors of lesion recurrence and retreatment.CONCLUSION: There is a significant difference in the initial cure effect between the two drugs in Zone II type 1 ROP, with the reactivation and retreatment rates of the IVC group being much lower than those of the IVR group.

Objective To explore the role of mitochondrial autophagy in ovarian inflammation associated with poly-cystic ovary syndrome(PCOS)based on the NOD-like receptor thermoprotein domain-related protein 3(NLRP3)pathway.Methods Human ovarian granulosa cell line SVOG was treated with 25 nmol/L dihydrotestosterone(DHT)for 24 h to establish PCOS cell model.SVOG cells were transfected with adenovirus carrying NLRP3(Ad-NLRP3)and negative vector(Ad-EV)or NLRP3 shRNA(sh-NLRP3)and negative control(sh-NC)to overex-press or knockdown NLRP3.Mito-Tracker staining and GFP-LC3 staining were used to evaluate mitochondrial auto-phagy in cells.TUNEL staining,JC-1 staining and Mito-SOX staining were used to analyze the apoptosis,mito-chondrial membrane potential and mitochondrial-derived superoxide production.32 female BALB/c mice were ran-domly divided into three groups:control(Con)group,DHEA group,DHEA+sh-NC group and DHEA+sh-NL-RP3 group,with 8 mice in each group.Except the control group,all other groups treated mice with dehydroepi-androsterone(DHEA)to establish PCOS mouse model.DHEA+sh-NLRP3 group and DHEA+sh-NC group were administrated with sh-NLRP3 or sh-NC encapsulated in lentivirus at a concentration of 1 x 109 TU/ml via tail vein injection.The ultrastructure of mitochondria in ovarian tissue of mice in each group was observed by transmission e-lectron microscope.Results Compared with DHT+sh-NC group,the level of NLRP3 of SVOG cells in DHT+sh-NLRP3 group decreased(P＜0.05).The co-location of GFP-LC3 and mitochondria in SVOG cells in DHT+sh-NLRP3 group was higher than that in DHT+sh-NC group(P＜0.05).Compared with DHT+sh-NC group,the number of TUNEL positive cells and Mito-SOX fluorescence density of SVOG cells in DHT+sh-NLRP3 group de-creased,and the ratio of polymer JC-1 to monomer JC-1 increased(P＜0.05).Compared with Con+Ad-EV group,the level of NLRP3,the number of TUNEL-positive cells and the fluorescence density of mito-SVOG in Con+Ad-NLRP3 group increased(P＜0.05),and the co-location level of GFP-LC3 and mitochondria decreased;the ratio of polymer JC-1 to monomer JC-1 decreased(P＜0.05).Compared with the control group,TUNEL positive cells,relative ROS intensity and percentage of damaged mitochondria in the ovarian tissue of mice in DHEA group increased(P＜0.05).Compared with DHEA+sh-NC group,TUNEL positive cells,relative ROS intensity and percentage of damaged mitochondria in DHEA+SH-NLRP group decreased(P＜0.05).Conclusion Inhibition of mitochondrial autophagy induced by activation of NLRP3 leads to mitochondrial dysfunction and promotes mito-chondrial-related apoptosis in GCs.Knockdown of NLRP3 is beneficial to mitochondrial homeostasis and improves the resistance of GCs to oxidative stress injury,thus promoting the recovery of PCOS.

OBJECTIVE To study the toxicokinetics and tissue distribution characteristics of alpha-amanitin in rats.METHODS The tail venous blood was collected from SD rats before and 5,10,20,30 and 45 min,1,1.5,2.5,4 and 8 h after intraperitoneal injection of alpha-amanitin(1.5 mg·kg-1),and the concentration of alpha-amanitin in blood was determined by liquid chromatography-mass spectrometry(LC-MS/MS).DAS 2.0 software was used to analyze and plot the drug-time curve with toxicokinetic parame-ters.Based on the toxicokinetics results,18 SD rats were randomly divided into three groups.The rats were sacrificed,and left ventricular arterial(LVA)blood and 9 types of tissue samples involving the heart,liver,spleen,lung,kidney,whole brain,small intestine,stomach wall and testis were collected 15 min,40 min and 2.5 h after dosing,and the concentrations of alpha-amanitin were measured by LC-MS/MS to obtain the tissue distribution results of alpha-amanitin in SD rats.RESULTS Toxicokinetics studies revealed that the peak blood concentration(Cmax)was(633±121)μg·L-1,the elimination half-life(T1/2)was(0.72±0.37)h,and the peak time(Tmax)was(0.52±0.16)h.The total clearance rate(CLz)was(1.62±0.26)L·h·kg-1,the area under the curve(AUC0-t)was(946±183)μg·h·L-1,and the mean reten-tion time(MRT0-t)was(1.18±0.17)h.The apparent volume of distribution(Vz)was(1.65±0.86)L·kg-1.The results of tissue distribution study showed that alpha-amanitin was widely distributed in SD rats with the highest concentration in the kidney,followed by the lung,small intestines,stomach wall,LVA blood and liver,but was low in the heart,spleen,testicles and other tissues,and very low in the brain.Alpha-amanitin was absorbed and eliminated quickly,peaked at 40 min in each tissue,and the concen-tration was minimized after 2.5 h.CONCLUSION The absorption and elimination of alpha-amanitin by intraperitoneal injection are rapid in SD rats,and the blood concentration reaches the peak about 31 min after administration,but can not be detected 4 h later.Alpha-amanitin is mainly distributed in the kidney,followed by the tissues and metabolic organs with rich blood flow,such as the lung,small intestines,stomach wall,LVA blood and liver.The content of alpha-amanitin is low in the heart,spleen,testicles and other tissues,and very low in the brain.It is speculated that it may have toxic targeting effect on the kidney and low blood-brain barrier permeability.

Objective To investigate the value of electrocardiographic parameters in assessing severe coronary artery lesions in patients with coronary artery disease.Methods A total of 364 patients with coronary heart disease admitted to the Second Affiliated Hospital,Zhengzhou University from January 2022 to September 2023 were selected as the subjects.The patients were divided into severe lesion group(n=123)and non-severe lesion group(n=241)based on the Gensini score.The two groups were compared in terms of baseline data and electrocardiographic parameters.Logistic regression was used to analyze the factors affecting severe lesions and the efficacy of ECG parameters assessment.A column-line plot of ECG parameters to assess severe lesions was created.The joint incremental value of ECG parameters and the area under the ROC curve for assessing severe lesions were calculated.Results The patients in the severe lesion group were elder in age and they showed significantly higher rates in creatine kinase isoenzyme,amino-terminal brain natriuretic peptide precursor,QRS time frame,frontal QRS-T pinch angle,smoking,hypertension,diabetes mellitus,pathologic Q waves,left ventricular hyper voltage,fragmented QRS waves,and significantly lower rates in low HDL cholesterol and ejection fraction,as compared to the non-severe lesion group(all P<0.05).Logistic regression analysis showed that pathological Q waves(OR=3.71,95%CI:1.93～7.15,P<0.001),left ventricular hyper voltage(OR=5.05,95%CI:1.66～15.41,P=0.004),fragmented QRS waves(OR=2.17,95%CI:1.14～4.11,P=0.018),and frontal QRS-T pinch angle(OR=1.01,95%CI:1.00～1.02,P=0.023)were the factors for assessing severe lesions.The incremental value of combined ECG parameters for severe lesions:C-statistic 0.09(95%CI:0.02～0.16,P=0.009),NRI 0.37(95%CI:0.20～0.55,P<0.001),and IDI 0.12(95%CI:0.08～0.17,P<0.001).The ROC curves showed an AUC of 0.79(95%CI:0.74～0.84,P<0.001),a sensitivity of 63.41%,and a specificity of 85.89%for the combined assessment of severe lesions by ECG parameters.Conclusion The combined observations of electrocardiographic parameters such as pathologic Q waves,left ventricular hyper voltage,fragmented QRS waves,and frontal QRS-T pinch angle have an affirmativeclinical value in the assessment of severe lesions.

Objective:Rejection remains the most important factor limiting the survival of transplanted kidneys.Although a pathological biopsy of the transplanted kidney is the gold standard for diagnosing rejection,its limitations prevent it from being used as a routine monitoring method.Recently,peripheral blood lymphocyte subpopulation testing has become an important means of assessing the body's immune system,however,its application value and strategy in the field of kidney transplantation need further exploration.Additionally,the development and utilization of routine test parameters are also important methods for exploring diagnostic strategies and predictive models for kidney transplant diseases.This study aims to explore the correlation between peripheral blood lymphocyte subpopulations and T cell-mediated rejection(TCMR)and antibody-mediated rejection(ABMR),as well as their diagnostic value,in conjunction with routine blood tests. Methods:A total of 154 kidney transplant recipients,who met the inclusion and exclusion criteria and were treated at the Second Xiangya Hospital of Central South University from January to December,2021,were selected as the study subjects.They were assigned into a stable group,a TCMR group,and an ABMR group,based on the occurrence and type of rejection.The basic and clinical data of these recipients were retrospectively analyzed and compared among the 3 groups.The transplant kidney function,routine blood tests,and peripheral blood lymphocyte subpopulation data of the TCMR group and the ABMR group before rejection treatment were compared with those of the stable group. Results:The stable,TCMR group,and ABMR group showed no statistically significant differences in immunosuppressive maintenance regimens or sources of transplanted kidneys(all P>0.05).However,the post-transplant duration was significantly longer in the ABMR group compared with the stable group(P<0.001)and the TCMR group(P<0.05).Regarding kidney function,serum creatinine levels in the ABMR group were higher than in the stable group and the TCMR group(both P<0.01),with the TCMR group also showing higher levels than the stable group(P<0.01).Both TCMR and ABMR groups had significantly higher blood urea nitrogen levels than the stable group(P<0.01),with no statistically significant difference between TCMR and ABMR groups(P>0.05).The estimated glomerular filtration rate(eGFR)was lower in both TCMR and ABMR groups compared with the stable group(both P<0.01).In routine blood tests,the ABMR group had lower hemoglobin,red blood cell count,and platelet count than the stable group(all P<0.05).The TCMR group had higher neutrophil percentage(P<0.05)and count(P<0.05)than the stable group,and the ABMR group had a higher neutrophil percentage than the stable group(P<0.05).The eosinophil percentage and count in the TCMR group were lower than in the stable and ABMR groups(all P<0.05).Both TCMR and ABMR groups had lower basophil percentage and count,as well as lower lymphocyte percentage and count,compared with the stable group(all P<0.05).There were no significant differences in monocyte percentage and count among the 3 groups(all P>0.05).In lymphocyte subpopulations,the TCMR and ABMR groups had lower counts of CD45+cells and T cells compared with the stable group(all P<0.05).The TCMR group also had lower counts of CD4+T cells,NK cells,and B cells than the stable group(all P<0.05).There were no significant differences in the T cell percentage,CD4+T cell percentage,CD8+T cell percentage and their counts,CD4+/CD8+T cell ratio,NK cell percentage,and B cell percentage among the stable,TCMR,and ABMR groups(all P>0.05). Conclusion:The occurrence of rejection leads to impaired transplant kidney function,accompanied by characteristic changes in some parameters of routine blood tests and peripheral blood lymphocyte subpopulations in kidney transplant recipients.The different characteristics of changes in some parameters of routine blood tests and peripheral blood lymphocyte subpopulations during TCMR and ABMR may help predict and diagnose rejection and differentiate between TCMR and ABMR.

This study was aimed at analyzing the Salmonella infection rate,serotype distribution,and drug resistance rate among children with foodborne diseases in Wuhan,to provide evidence for the prevention and control of foodborne diseases in children.We collected pediatric cases of foodborne diseases and fecal specimens in Wuhan from 2017 to 2022,and conducted statistical analysis in SPSS 22.0.A total of 1 180 cases of foodborne diseases in children were monitored,and 281 strains of Salmonella were detected.Salmonella was distributed across 27 serotypes,primarily Salmonella enteritidis(138 strains,ac-counting for 49.11％)and Salmonella typhimurium(37 strains,accounting for 13.17％).The Salmonella-positive cases in-cluded more boys than girls(1.58∶1 ratio),and the age group with the highest detection rate was 7-18 years(34.48％).The proportion of positive patients gradually decreased,and the detection rate gradually increased,with increasing age.Milk and dairy products,fruits and fruit products,and grain and grain products were the main suspected food exposures with high Sal-monella detection rates.The Salmonella detection rate was highest in the third quarter(44.64％),and significant differences were observed in the positivity rates of Salmonella samples across seasons(x2=178.483,P＜0.05).The main clinical mani-festations of children with Salmonella infection were fever(87.19％)and diarrhea(96.44％),primarily watery stool(51.96％).Salmonella showed different degrees of resistance to 14 antibiotics(1.08％-75.99％),primarily ampicillin(75.99％),tetracycline(68.10％),and cefazolin(50.18％).A significant seasonal pattern was observed in the detection of Sal-monella in foodborne diseases among children in Wuhan,and in various suspected food exposures.The serotype composition was polymorphic,and the Salmonella resistance rate was relatively high.Therefore,active monitoring of foodborne diseases and Salmonella resistance surveillance must continue to be strengthened.

On the basis of the clinical characteristics of atherosclerosis(AS)in traditional Chinese medicine(TCM)and Western medicine,this paper analyzes common animal models of AS.The coincidence of clinical characteristics of the models was scored in the hope of providing new ideas and a reference for those studying AS.This paper reviews the varieties,modeling method,modeling principles,and characteristics of common animal models of AS.Moreover,similarities among common animal models,in terms of their clinical diagnostic criteria and symptom characteristics,were assessed.High-fat feeding type,mechanical injury combined with high-fat feeding type,genetic engineering combined with high-fat feeding type,chemical induction combined with high-fat feeding type,and combined Chinese clinical syndrome and Western disease AS models are widely established.Comparative analysis showed that balloon injury combined with high fat feeding type,ApoE receptor-knockout mouse combined with high-fat diet type,and phlegm and blood stasis type models of disease and symptom combinations showed a comparatively high level of clinical agreement between Chinese and Western medicine.Presently,most animal models of AS have a high degree of relevance to Western medicine,and the evaluation criteria used for the models are predominately from a Western medicine perspective.Models that combine disease and syndrome are lacking,hindering the development of wholism concepts and treatment through the differentiation of syndromes used in TCM.Therefore,establishing an animal model with a high degree of accuracy and coincidence between TCM and Western perspectives that combines the disease and its TCM symptoms is a top priority for studying the prevention and treatment of AS.

Purpose To investigate the clinicopathologic,immunophenotypic features,genetic alterations and prognosis of melanotic Xp11 neoplasms/melanotic TFE3-rearrangement neo-plasms.Methods Five cases were selected from the Depart-ment of Pathology,Union Hospital,Huazhong University of Sci-ence and Technology from November 2018 to July 2023.The clinicopathologic,immunohistochemical,FISH assays,next-generation sequencing(NGS)and follow-up details were collect-ed.Results There were 1 male and 4 females,with their ages ranging from 16 to 59 years(mean 28.2 years).The maximum diameters of the masses were 3-6 cm(average 4.7 cm).The tumors located in right kidneys(3 cases),tubal interstitium(1 case)and pelvis(1 case).Microscopically,most tumors shared similar morphology such as nested,acinar structures sep-arated by a delicate vascular network.Epithelioid tumor cells presented with clear to granular eosinophilic cytoplasm.Lym-phocytic infiltration was seen in the background;melanin depo-sition was noted in the cases;neoplastic necrosis was detected in 4 cases.Mitotic activity was low with 5 cases showing＜3/10 HPF.Intravascular tumor thrombus was detected in 2 cases,no lymphovascular and nerve invasions were detected in other 3 ca-ses.Immunohistochemically,all 5 cases expressed TFE3 dif-fusely,and expressed HMB45,Melan A to varying degrees,CK(AE1/AE3),CK7,EMA,PAX8,TFEB,S-100,SOX10,SMA,desmin were all non-reactive in the 5 cases.The Ki67-la-beling index was＜20％.TFE3 separation signal in 4 cases were detected by FISH,1 case was interpreted as negative due to atypical signal which was confirmed by next-generation se-quencing(NGS)assay as RBM10-TFE3.Clinical follow-up was available for five patients for 2-60 months,in which four pa-tients were alive with no evidence of disease after initial resec-tion,and one patient with thoracic spine metastasis was currently in stable condition.Conclusion Melanotic Xp1 1 neoplasms/melanotic TFE3-rearrangement neoplasms has unique morpholog-ic,immunophenotypic and genetic characteristics.It might be reclassified into a distinctive malignant mesenchymal tumor enti-ty.

Purpose To explore the value of contrast-enhanced ultrasound(CEUS)in the differential diagnosis of gallbladder polypoid lesions(GPLs)(diameter≥10 mm).Materials and Methods A prospective enrollment of 229 patients with GPLs who underwent cholecystectomy in 17 hospitals from December 1 2021 to June 30 2024 was conducted to analyze the relationship between general data,conventional ultrasound,CEUS characteristics and the nature of GPLs.Multivariate Logistic regression was employed to identify independent risk factors for neoplastic polyps,the differential diagnostic value of different indicators was compared.Results Among 229 patients with GPLs,there were 108 cases of cholesterol polyps,102 cases of adenoma and 19 cases of gallbladder cancer.Age(Z=-4.476,P<0.001),polyp number(χ2=15.561,P<0.001),diameter(Z=-8.149,P<0.001),echogenicity(χ2=9.241,P=0.010),vascularity(χ2=23.107,P<0.001),enhancement intensity(χ2=47.610,P<0.001),enhancement pattern(χ2=6.468,P=0.011),vascular type(χ2=84.470,P<0.001),integrity of gallbladder wall(χ2=7.662,P=0.006)and stalk width(Z=-9.831,P<0.001)between cholesterol polyps and neoplastic polyps were statistically significant.Age,location,diameter,echogenicity,enhancement pattern,vascular type and stalk width between adenoma and gallbladder cancer were statistically significant(Z=-4.333,-3.902,-5.042,all P<0.05).Multivariate Logistic regression analysis showed that hyper-enhancement,branched vascular type and stalk width were independent risk factors for neoplastic polyps(OR=4.563,5.770,3.075,all P<0.001).The combination of independent risk factors was better than single factor and diameter in the differential diagnosis of cholesterol polyps and neoplastic polyps(all P<0.01).Conclusion CEUS can effectively identify the nature of GPLs and provide a valuable imaging reference for the selection of treatment methods.