Objective: To report the antibody identification, blood management during pregnancy and the monitoring process of fetal hemolytic disease of fetus and newborn (HDFN) in a pregnant woman with a history of blood transfusion and pregnancy who developed anti-Jr . Methods: Saline tube technique and anti-human globulin technique were used for maternal blood typing, unexpected antibody screening and identification, as well as for determining antibody titer and IgG subclasses. PCR-SSP was employed for genotyping of 18 blood group systems. Next-generation sequencing (NGS) was utilized for gene sequencing of 38 blood group systems. Sanger sequencing was applied to verify rare blood group mutations detected by NGS and to investigate the corresponding rare blood group genes in family members. Blood preparation was achieved through anemia management in prenatal clinics and autologous blood collection during pregnancy. The newborn underwent the three primary tests for HDFN and plasma IgG subclass testing. Results: The pregnant woman's blood type was B, RhD positive, with a positive unexpected antibody screen, and the antibody identification pattern was consistent with a high-frequency antigen antibody. Gene sequencing revealed a homozygous ABCG2 c.376C>T mutation in the woman, resulting in the Jr(a-) phenotype, and anti-Jr antibody was present in her plasma. No compatible Jr(a-) blood was found among family members. The maternal anti-Jr IgG titer remained stable at 256 during pregnancy, with no detectable IgG1 or IgG3 subclasses against the Jr antigen. A total of 800 mL of autologous blood was collected in two stages during pregnancy. The newborn was B, RhD positive, Jr(a+), with a positive unexpected antibody screen (anti-Jr ). IgG subclass typing detected no IgG1 or IgG3. The direct antiglobulin test was positive, while the acid elution test was negative. Conclusion: The combination of serology and blood group genetic analysis provides a diagnostic basis for identifying antibodies to high-frequency antigens. Managing perinatal anemia and implementing staged autologous blood storage can secure blood supply for the perioperative period. IgG antibody subclass typing offers a reference for clinical assessment and prevention of HDFN.

Abstract Childhood hypertension is becoming a substantial public health challenge with profound implications for children s quality of life and long term health. The study analyzes the global prevalence of childhood hypertension and the relationship between macroecological factors, meso environmental factors, and micro individual factors based on the perspective of life course and childhood hypertension. And it further summarizes existing prevention and control strategies: systematic prevention and control based on policy and social support, health promotion based on behavioral science theory, and dynamic monitoring and management based on individualized prevention and control, to provide a reference for promoting the advancement of childhood hypertension prevention and control strategies.

ObjectiveTo investigate the triglyceride-glucose (TyG) index and the level of serum homocysteine (Hcy) in association with the incidence of stroke in type 2 diabetes mellitus (T2DM) patients. MethodsBased on the chronic disease risk factor surveillance cohort in Pudong New Area, Shanghai, excluding those with stroke in baseline survey, T2DM patients who joined the cohort from January 2016 to October 2020 were selected as the research subjects. During the follow-up period, a total of 318 new-onset ischemic stroke patients were selected as the case group, and a total of 318 individuals matched by gender without stroke were selected as the control group. The Cox proportional hazards regression model was used to adjust for confounding factors and explore the serum TyG index and the Hcy biochemical indicator in association with the risk of stroke. ResultsThe Cox proportional hazards regression results showed that after adjusting for confounding factors, the risk of stroke in T2DM patients with 10 μmol·L⁻¹-¹ and Hcy>15 μmol·L⁻¹ was 1.532 times (95%CI: 1.017‒2.309 times, P=0.041) and 1.738 times (95%CI: 1.119‒2.699 times, P=0.014) higher respectively, compared to T2DM patients with Hcy≤10 μmol·L⁻¹. T2DM patients with TyG>9.074 had 1.396 times higher risk of stroke (95%CI: 1.091‒1.787, P=0.008) compared to those with TyG≤9.074. The study found that urea and α1-antitrypsin (α1-AT) were independent risk factors for the incidence of stroke in T2DM patients. For every unit increase in urea andα1-AT, the risk of stroke in T2DM patients increased by 233.6% (HR=3.336, 95%CI: 1.588‒7.009, P=0.001) and 11.3% (HR=1.113, 95%CI: 1.028‒1.205, P=0.008), respectively. Cumulative risk curves showed that Hcy>10 μmol·L⁻¹ and TyG>9.074 accelerated the time to stroke occurrence in T2DM patients. ConclusionElevated levels of Hcy>10 μmol·L⁻¹ and a higher TyG index are risk factors for stroke in T2DM patients. Effective interventions for Hcy and TyG should be conducted for diabetic patients to reduce the incidence of stroke.

Vaccine immunization is the most effective and cost-efficient method for infectious disease prevention and control.Since the outbreak of novel coronavirus pneumonia(caused by the novel coronavirus,COVID-19)at the end of 2019,third generation mRNA nucleic acid vaccines has been applied to stop viral spread.mRNA vaccines,rather than relying on the immune activation mode of traditional vaccines,are an innovative breakthrough using the body's own cells to produce antigens,thereby activating double specific immunity,forming immune memory,and providing more lasting specific immunity.Com-pared with the traditional first-generation(inactivated)and second-generation(genetically engineered)vaccines,mRNA vac-cines,because of the advantages provided by this platform,play important roles in the prevention and control of major sudden infectious diseases.Consequently,mRNA vaccines were the world's first COVID-19 vaccines to be applied clinically,thus ser-ving as a barometer in the field of vaccine research and development.Herein,the molecular design and presentation of mRNA vaccines,and the molecular mechanisms of their activation of the immune response are reviewed,to provide a theoretical basis for future application of novel mRNA vaccines in the prevention and control of animal infectious diseases.

Objectives:This study aims to investigate the incidence,characteristics and clinical outcomes of vagal response(VR)during pulmonary vein isolation(PVI).Methods:A total of 702 patients with nonvalvular atrial fibrillation(AF)who received the first PVI radiofrequency ablation in Central China Fuwai Hospital from January 2022 to December 2023 were consecutively enrolled.PVI was initiated from right superior pulmonary vein(RSPV),followed by other pulmonary veins(PVs).The VR was defined as atrioventricular block(AVB),asystole or a 50%increase in the RR interval.Results:Among 702 patients with AF,380 patients(54.1%)were paroxysmal AF and 322 patients(45.9%)were persistent AF.77 patients(11.0%)developed 81 VR episodes,which were more common in paroxysmal group than in persistent AF group(19.5%vs.0.9%,P<0.001).VR manifestations included 51 sinus arrest(63.0%),26 sinus bradycardia(32.1%),and 4 atrioventricular block(AVB,4.9%).Most VR episodes were observed in the left superior ganglionated plexi(67[82.7%]).Paroxysmal AF(OR=18.667,95%CI:6.638-52.491,P<0.001),body mass index(BMI)≥28.0 kg/m2(OR=2.361,95%CI:1.376-4.051,P=0.002)and left ventricular ejection fraction(LVEF)≥62.0%(OR=1.964,95%CI:1.119-3.447,P=0.019)were independent risk factors of VR.During a mean of(13.0±7.1)months follow up,among paroxysmal AF patients,6 patients(8.1%)with VR and 33 patients(10.8%)without VR experienced AF recurrence(P=0.496).Kaplan-Meier curves estimated that the AF-free survival rate was similar between VR group and non-VR group among paroxysmal AF patients(log-rank P=0.735).Conclusions:The most common sites of VR when initiating PVI from right RSPV occur in left superior ganglionated plexi.Paroxysmal AF,BMI≥28.0 kg/m2 and LVEF≥62.0%are independent risk factors of VR.VR does not affect AF-free survival.

Objective To explore the mechanism of IgA nephropathy(IgAN)caused by multi-glycosides of Tripterygium wilfordii(GTW)through the regulation of Silent information regulatory factor 1(SIRT 1)/nuclear transcription factor E2-related factor 2(Nrf 2)/antioxidant enzyme heme oxygenase 1(HO-1)signaling pathway.Methods Forty-five male SD rats were selected and randomly divided into two groups:the blank group(n=9)and the model group(n=36).In addition to the blank group,the BSA+CCl4+LPS group was used.At the end of 12 weeks,two rats were randomly selected for verification,and the model was successfully established.The 34 model rats were randomly divided into 3 groups:the model group(n=10),prednisone group(n=12),and GTW group(n=12).Urine,blood and kidney tissues were harvested 4 weeks after drug administration.Urinary erythrocyte number,24-h urinary protein quantification(24 h-UTP),alanine transaminase(ALT),serum albumin(ALB),urea nitrogen(BUN),and blood creatinine(SCr)were performed for each group;the protein expression of SIRT1,Nrf2,HO-1 and PINK1 was detected by Western blotting analysis;real-time polymerase chain reaction(RT-PCR)detection of SIRT1,Nrf2,HO-1 and PINK1 mRNA expression in rat kidney tissue;and detection of IgA deposition in the renal mesangial area by immunofluorescence.Kidney histopathological changes were observed in all the rats by hematoxylin-eosin(HE)staining.Results The results compared with those in the blank group,the urinary red blood cell count and 24 h-UTP,ALT,BUN,and SCr levels were significantly greater(P<0.01);The ALB level was significantly lower(P<0.01);renal tissue SIRT1,Nrf2,HO-1,PINK1 protein and mRNA expression were significantly lower(P<0.01);IgA deposition in the mesentery was obvious;renal pathological damage was severe;and the difference was statistically significant. Compared with those in the model group,urinary red blood cell counts and 24 h-UTP,ALT,BUN,and SCr levels in the prednisone and GTW groups were significantly lower (P<0.01);ALB levels were significantly greater (P<0.01);SIRT1,Nrf2,HO-1,PINK1 protein and mRNA expression were significantly greater (P<0.01);IgA deposition in the mesangial area was reduced,and renal pathology was improved,with statistically significant difference. Conclusions GTW may alleviate oxidative stress injury,protect renal function,and improve renal injury by activating the SIRT 1/Nrf 2/HO-1 signaling pathway.

Objectives:To analyze the disease burden and trends of ischemic stroke attributable to major metabolic risk factors in China from 1990 to 2021,and provide a basis for disease prevention and management.Methods:Based on the Global Burden of Disease(GBD)2021 data,we compared the age-standardized mortality rate(ASMR)and age-standardized disability-adjusted life year rate(ASDR)of ischemic stroke attributable to major metabolic risk factors(including overweight/obesity,hypertension,elevated low-density lipoprotein cholesterol,low bone density,hyperglycemia,and impaired renal function)in China and globally,and estimated their time trends through estimated annual percentage change(EAPC).We also analyzed the number of deaths,mortality rate,disability adjusted life year(DALY),and DALY rate of ischemic stroke attributable to major metabolic risk factors in China from 1990 to 2021,and compared the epidemiological differences among different gender and age groups.Results:From 1990 to 2021,the disease burden of ischemic stroke attributable to major metabolic risk factors in China showed a slight upward trend(1990 to 2005)and then gradually decreased,with a relatively small overall decline.,while the global disease burden showed a decreasing trend.In 2021,the ASMR and ASDR of ischemic stroke attributable to major metabolic risk factors in China were 48.11/100 000 and 898.35/100 000(EAPC[95%CI]were-0.24%[-1.13%to 0.66%]and-0.25%[-1.00%to 0.51%]),respectively,both higher than the global average levels(33.96/100 000 and 651.46/100 000,EAPC[95%CI]were-1.75%[-2.03%to 1.47%]and-1.49%[-1.74%to-1.23%],respectively).Compared with 1990,the actual mortality rate and DALY rate of ischemic stroke attributable to major metabolic risk factors in China increased by 139.60%and 110.53%respectively in 2021.There are gender differences in the disease burden of ischemic stroke attributable to major metabolic risk factors in China,with significantly higher death numbers,mortality rates,DALY,and DALY rates in males compared to females.At the age level,DALY and DALY rates were both higher in individuals aged 15-49,DALY increased but DALY rates decreased in individuals aged 50-69.DALY surged in individuals aged 70 and above,male DALY rates showed increasing trend and female DALY rates showed slightly decreasing trend.Conclusions:The disease burden of ischemic stroke in China is significantly affected by metabolic risk factors.Although prevention and control have achieved certain positive results,the overall disease burden in China is higher than that of the world,and precise intervention strategies need to be developed for different genders and ages in China.

Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Objective To investigate the relationship between plasma von willebrand factory-lytic protease (ADAMTS13) level and the severity of the disease and its prognostic value in patients with endometriosis (EMS). Methods 130 patients with EMS treated in the hospital from January 2019 to December 2022 were retrospectively selected. Plasma ADAMTS13 level was detected by enzyme-linked immunosorbent assay (ELISA),and its correla-tion with American Fertility Society (r-AFS) stage and endometriosis Health Status Scale (EHP-30) score was analyzed. The 1-year prognosis was recorded,and the risk factors of poor prognosis were analyzed by logistic regression. Receiver operating characteristic curve (ROC) was plotted to evaluate the predictive value of plasma ADAMTS13 levels in patients with EMS. Results The plasma ADAMTS13 level of EMS patients decreased signifi-cantly with the increase of r-AFS stage,and was negatively correlated with the score of EHP-30 (P<0.001). Multiple factors showed that low plasma ADAMTS13 level,bilateral location of pelvic lesions and high r-AFS stage were independent risk factors for poor prognosis (all P<0.05). ROC analysis showed that the area under the curve (AUC) of plasma ADAMTS13 was 0.889 (cut-off point 13.315,sensitivity 0.931,specificity 0.690). Conclusion Plasma ADAMTS13 level is negatively correlated with the severity of EMS,and is an independent risk factor for poor prognosis of EMS patients,and has a certain predictive value for prognosis of EMS patients.

Objective To construct a model of knee osteoarthritis(KOA)through the co-culture of dorsal root ganglia(DRG)and fibroblast-like synoviocytes(FLSs).To investigate the effects of transient receptor potential vanilloid type 1(TRPV1)desensitization of sensory neurons induced by mechanical stimulation,including the alleviation of the FLSs inflammatory response.Methods DRG neuronal cells were identified through immunofluorescence.The stress loading of DRG neurons was realized using the FX-6000T cell stress system,and the effect of mechanical strain on the activity of DRG neurons was measured using the CCK-8 method.Ca2+ion flux in DRG neurons was studied through flow cytometry.A Transwell chamber and FLSs were used to establish a co-culture system.The contents of the pro-inflammatory factors IL-1β,TNF-α,and TGF-β in the supernatant were determined by ELISA.Gene and protein expression levels of TRPV1 and its desensitizing negative regulatory proteins PP2B,CaM,IL-1β,TNF-α,TGF-β,and α-SMA in DRG neurons were evaluated using RT-qPCR and Western blot,respectively.Results The Ca2+ion flux in DRG neurons increased under inflammatory conditions,and low intensity(sinusoidal,2％,1 Hz,6 h)thumb-pressing-induced mechanical stimulation did not alter Ca2+ion flux(P＞0.05).However,middle intensity(sinusoidal,4％,1 Hz,6 h)and high intensity(sinusoidal,8％,1 Hz,6 h)stimulation increased Ca2+ion flux significantly(P＜0.05).Notably,high intensity stimulation did not lead to a further increase in Ca2+ion flux over that for middle intensity stimulation(P＞0.05).There was no significant effect of low intensity stimulation on TRPV1,PP2B,or CaM gene or protein expression in DRG neurones,on IL-1β,TNF-α,or TGF-βconcentrations in the supernatant of co-cultured cells,or on IL-1β,TNF-α,TGF-β,or α-SMA gene or protein expression in FLSs(P＞0.05).Middle and high intensity mechanical stimulation up-regulated TRPV1 at the gene and protein expression levels in DRG neurons in the inflammatory group(P＜0.05)but down-regulated PP2B and CaM at the gene and protein expression levels(P＜0.05).Middle and high intensity mechanical stimulation decreased IL-1β,TNF-α,and TGF-β levels in the supernatants of co-cultured cells(P＜0.05)and decreased the gene and protein expression levels of IL-1β,TNF-α,TGF-β,and α-SMA in FLSs(P＜0.05).Conclusions Middle and high intensity thumb-pressing-induced mechanical stimulation induced TRPV1 desensitization of rat sensory neurons,reduced the release of pain mediators,and suppressed the FLSs inflammatory response through downregulating IL-1β,TNF-α,and TGF-β.