Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

ObjectiveBy exploring the influencing factors of readmission in patients with stable angina of coronary heart disease （CHD） based on real-world clinical data， to establish a risk prediction model of integrated traditional Chinese and western medicine， in order to provide a basis for early identification of high-risk populations and reducing readmission rates. MethodsA prospective clinical study was conducted involving patients with stable angina pectoris of CHD， who were divided into a training set and a validation set at a 7∶3 ratio. General information， traditional Chinese medicine （TCM）-related data， and laboratory test results were uniformly collected. After a one-year follow-up， patients were classified into a readmission group and a non-readmission group based on whether they were readmitted. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for readmission. A risk prediction model of integrated traditional Chinese and western medicine was constructed and visualized using a nomogram. The model was validated and evaluated in terms of discrimination， calibration， and clinical decision curve analysis. ResultsA total of 682 patients were included， with 477 in the training set and 205 in the validation set， among whom 89 patients were readmitted. Multivariate logistic regression analysis identified heart failure history ［OR = 6.93， 95% CI （1.58， 30.45）］， wiry pulse ［OR = 2.58， 95% CI （1.42， 4.72）］， weak pulse ［OR = 3.97， 95% CI （2.06， 7.67）］， teeth-marked tongue ［OR = 4.38， 95% CI （2.32， 8.27）］， blood stasis constitution ［OR = 2.17， 95% CI （1.06， 4.44）］， phlegm-stasis mutual syndrome ［OR = 3.64， 95% CI （1.87， 7.09）］， and elevated non-high-density lipoprotein cholesterol ［OR = 1.30， 95% CI （1.01， 1.69）］ as influencing factors of readmission. These factors were used as predictors to construct a nomogram-based risk prediction model for readmission in patients with stable angina. The model demonstrated moderate predictive capability， with an area under the receiver operating characteristic curve （AUC） of 0.818 ［95% CI （0.781， 0.852）］ in the training set and 0.816 ［95% CI （0.779， 0.850）］ in the validation set. The Hosmer-Lemeshow test showed good calibration （χ² = 4.55， P = 0.80）， and the model's predictive ability was stable. When the threshold probability exceeded 5%， the clinical net benefit of using the model to predict readmission risk was significantly higher than intervening in all patients. ConclusionHistory of heart failure， teeth-marked tongue， weak pulse， wiry pulse， phlegm-stasis mutual syndrome， blood stasis constitution， and non-high-density lipoprotein cholesterol are influencing factors for readmission in patients with stable angina of CHD. A clinical prediction model was developed based on these factors， which showed good discrimination， calibration， and clinical utility， providing a scientific basis for predicting readmission events in patients with stable angina.

Porcine hemagglutinating encephalomyelitis virus(PHEV),a betacoronavirus with a sin-gle-stranded,positive-sense RNA genome,is an important pathogen in the swine industry.It causes porcine hemagglutinating encephalomyelitis(PHE),presenting with symptoms such as vomiting,diarrhea,neurological symptoms,and respiratory distress,particularly in piglets.The virus is asso-ciated with high mortality rates and growth stunting in subclinical cases.PHEV has a well-docu-mented global distribution,with occurrences reported across Europe,South America,North Amer-ica,and East Asia,posing a considerable challenge to the swine industry.This review aims to pro-vide a comprehensive overview of PHE's virological characteristics,epidemiology,clinical and pathological manifestations,mechanisms of infection and pathogenicity,and the current state of di-agnostic techniques.It further evaluates advancements in vaccine and antiviral development,along-side comprehensive prevention and control strategies,contextualized within the framework of the latest foundational research.

OBJECTIVE: To evaluate the clinical significance of a hemizygous c.1042-10G>C variant of the SLC9A7 gene NM_001257291.2) previously identified in individuals with neurodevelopmental disorders, and to provide an evidence-based guidance for prenatal genetic counseling. METHODS: Four families presented at the Medical Genetics Center of Henan Provincial People's Hospital between December 2022 and July 2024 were included in this study. Phenotypic information and biological samples were collected from family members. Genomic DNA was extracted and subjected to whole-exome sequencing and copy number variation analysis to identify candidate pathogenic variants. Sanger sequencing was performed for familial co-segregation analysis. Reverse-transcription PCR was used to assess the RNA splicing pattern of the variant in peripheral blood samples. Quantitative PCR was employed to analyze the expression profiles of various SLC9A7 transcripts in fetal brain tissue and peripheral blood samples. Pathogenicity of the variant was classified based on guidelines from the American College of Medical Genetics and Genomics (ACMG). This study was approved by the Medical Ethics Committee of Henan Provincial People's Hospital (Ethics No.: 2021-171). RESULTS: Six hemizygous males carrying the SLC9A7 c.1042-10G>C variant were identified among the four families, which included three adult males and two male infants with normal phenotypes. Only one affected male from family 3 exhibited global developmental delay, short neck, webbed neck, ocular dysplasia, and congenital corneal leukoma. He also had a history of perinatal asphyxia and carried an additional hemizygous variant HUWE1 c.12283C>G. Reverse-transcription PCR showed no aberrant splicing in heterozygous or hemizygous carriers compared to healthy controls, suggesting that the variant does not affect RNA splicing. Quantitative PCR revealed that NM_001257291.2 is the predominant transcript expressed in fetal brain tissue and peripheral blood. CONCLUSION The SLC9A7 c.1042-10G>C variant does not alter RNA splicing and is present in multiple phenotypically normal males, which supported its classification as a benign variant.
Humans ; Male ; Female ; Genetic Counseling ; Pedigree ; Adult ; DNA Copy Number Variations/genetics* ; Sodium-Hydrogen Exchangers/genetics* ; Exome Sequencing

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Objective:To explore the impacts of Yiqi Wenfei Tongqiao Formula on inflammatory response and T helper cell(Th)1/Th2 immune balance in rats with lung Qi deficiency cold type allergic rhinitis.Methods:Sixty SD rats were randomly divided into model group,low dose Yiqi Wenfei Tongqiao Formula group,medium dose Yiqi Wenfei Tongqiao Formula group,high dose Yiqi Wenfei Tongqiao Formula group,and desloratadine group,with 12 rats in each group,using a combination of smoking,cold stimula-tion combined with ovalbumin(OVA)sensitization and stimulation construct lung Qi deficiency cold type allergic rhinitis model.An-other 12 normal SD rats were selected as control group.Each group of rats was treated with corresponding drugs once a day for 14 days.Model group and control group rats were given physiological saline.Scoring method was used to analyze the behavioral score and lung index of rats with allergic rhinitis.HE staining was applied to detect pathological changes in rat nasal mucosa tissue.ELISA was ap-plied to detect levels of IFN-γ,IL-4 and TNF-α in rats serum;flow cytometry was applied to detect Th1/Th2.Western blot was applied to detect expressions of T-bet and GATA-3 protein in nasal mucosa tissues of rats in each group.Results:Compared with control group,pathological damage to nasal mucosa tissue of model group rats was severe,allergic rhinitis behavior score(days 41,48,56),lung index,levels of IL-4 and TNF-α,Th2 and protein expression of GATA-3 were increased,while levels of IFN-γ,Th1,Th1/Th2,and protein expression of T-bet were decreased(P<0.05);compared with model group,pathological damage to nasal mucosa tissue of rats in low,medium,high doses Yiqi Wenfei Tongqiao Formula groups and desloratadine group were alleviated,the allergic rhinitis behavior score(days 48,56),lung index,levels of IL-4 and TNF-α,Th2 and protein expression of GATA-3 were decreased,while levels of IFN-γ,Th1,Th1/Th2,and protein expression of T-bet were increased(P<0.05).Conclusion:Yiqi Wenfei Tongqiao Formula can inhibit inflammatory response of lung Qi deficiency cold type allergic rhinitis rats By regulating Th1/Th2 immune balance.

Objective:To investigate the gene expression of pulmonary metastatic papillary thyroid cancer (PTC) and its association with lung metastasis, facilitating risk assessment and personalized therapeutic strategies.Methods:A retrospective cohort study was conducted on 269 PTC patients (61 males, 208 females, age (38.9±11.9) years) treated at the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University from January 2017 to June 2020, including 60 cases with lung metastasis and 209 without. Primary tumor tissues underwent targeted next-generation sequencing (tNGS). Univariate and multivariate logistic regression analyses were performed to identify risk factors for lung metastasis, and χ2 test was used to evaluate the differences between the effective and ineffective groups of radioactive iodine (RAI) therapy. Results:In 60 PTC patients with lung metastasis, the mutation rates of B-Raf proto-oncogene, serine/threonine protein kinase (BRAF) V600E, telomerase reverse transcriptase (TERT) promoter, and rearranged in transformation (RET) fusion mutations were 28.3%(17/60), 25.0%(15/60), and 26.7%(16/60), respectively. Univariate analysis demonstrated that age≥55 years, multifocality, lateral cervical lymph node metastasis, extrathyroidal extension, BRAF V600E mutation, RET fusion, and TERT promoter mutation were significantly associated with lung metastasis in PTC ( Wald χ2 values: 4.13-31.28, all P<0.05). However, no significant statistical associations were observed between lung metastasis and gender, rat sarcoma type GTPase family (RAS) mutation, tumor protein p53 (TP53) mutation, phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) mutation, neurotrophic tyrosine receptor kinase 1 (NTRK1) fusion, or anaplastic lymphoma kinase (ALK) fusion ( Wald χ2 values: 0.01-3.50, all P>0.05). Multivariate analysis identified TERT promoter mutation (odds ratio ( OR)=11.86, 95% CI: 3.68-38.29, P<0.001), multifocality ( OR=5.30, 95% CI: 2.41-11.69, P<0.001), extrathyroidal extension ( OR=3.98, 95% CI: 1.77-8.98, P=0.001), and lateral cervical lymph node metastasis ( OR=3.13, 95% CI: 1.28-7.68, P=0.013) as independent risk factors for lung metastasis in PTC. Conversely, BRAF V600E mutation emerged as a potential protective factor ( OR=0.09, 95% CI: 0.04-0.21, P<0.001). The proportions of BRAF V600E mutation ( χ2=20.49, P<0.001) and TERT promoter mutation ( χ2=4.91, P=0.027) were higher in the RAI ineffective group. Conclusions:BRAF V600E mutation, TERT promoter mutation and RET fusion are related gene expression in lung metastasis of PTC. Multifocality, extrathyroidal extension, lateral cervical lymph node metastasis and TERT promoter mutation are risk factors for lung metastasis of PTC (TERT is an independent molecular risk marker), while BRAF V600E mutation may be a protective factor. RAI treatment efficacy of PTC patients with lung metastasis and BRAF V600E mutation/TERT promoter mutation is worse.

Objective To explore the effects of tripterygium glycosides(TGs)on adipocyte differentiation,inflam-mation,and fibrosis of orbital fibroblasts(OFs)in thyroid associated ophthalmopathy(TAO).Methods OFs isolated from 12 TAO patients were cultured and identified.CCK-8 experiment was used to detect the effect of different concentra-tions of TGs(0,12.5,25.0,50.0,100.0 mg·L-1)on the activity of OFs.Then,adipocyte differentiation phenotypes of OFs were induced and divided into a control group,an adipocyte differentiation group(DM),and TG groups(12.5,25.0,and 50.0 mg·L-1 TGs).Inflammation phenotypes of OFs were induced and divided into a control group,an IL-1 β group,and TG groups(12.5,25.0,and 50.0 mg·L-1 TGs).Fibrosis phenotypes of OFs were induced and divided into a control group,a TGF-βi group,and TG groups(12.5,25.0,and 50.0 mg·L-1 TGs).Oil red O staining was used to detect the formation of lipid droplets in OFs of adipocyte differentiation phenotype.ELISA was used to measure IL-6 and TNF-α levels in OFs of inflammation phenotype and HA levels in OFs of fibrosis phenotype.Transwell experiment was used to detect the cell migration rate of OFs of fibrosis phenotype.Real time quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the expression levels of PPAR-γ,c/EBP-α,FABP-4,perilipin-1,and adiponectin mRNAs and pro-teins in OFs of adipocyte differentiation phenotype,the expression levels of IL-6,TNF-α,COX-2,MCP-1 and ICAM-1 mR-NAs and proteins in OFs of inflammation phenotype,and the expression levels of Vimentin,Fibronectin,COL1A1,COL1A2 and ACTA2 mRNAs and proteins in OFs of fibrosis phenotype.Results The cells were identified as OFs.12.5,25.0,and 50.0 mg·L-1 of TGs had no significant effect on OF activity and these three concentrations were used in the subsequent ex-periment.In OFs of adipocyte differentiation phenotype,the number of orange red lipid droplets greatly decreased after TG treatment.The expression levels of PPAR-γ,c/EBP-α,FABP-4,perilipin-1,and adiponectin mRNAs and proteins in cells of TG groups were significantly lower than those of the DM group(all P＜0.05).In OFs of inflammation phenotype,IL-6 and TNF-α levels in supernatant and cells of TG groups were significantly decreased,compared with those of the IL-1βgroup(all P＜0.05).The expression levels of COX-2,MCP-1,and ICAM-1 mRNAs and proteins in cells of TG groups were significantly lower than those of the IL-1 β group(all P＜0.05).In OFs of fibrosis phenotype,the cell migration rates,HA levels in cell culture supernatant,and the expression levels of Vimentin,Fibronectin,COL1A1,COL1A2,and ACTA2 mR-NAs and proteins in cells of TG groups were significant lower than those of the TGF-β1 group(all P＜0.05).Conclusion TGs can inhibit the adipocyte differentiation,IL-1 β-induced inflammatory responses and TGF-β1-induced fibrosis of TAO-OFs.

Objective:To analyze the screening results of the Urban Cancer Early Diagnosis and Treatment Project in Qinghai Province from 2019 to 2024.Methods:A summary and statistical analysis were conducted on six years of screening data from the Urban Cancer Early Dia-gnosis and Treatment Program in Qinghai Province,with the high-risk rate,screening rate,and detection rate calculated separately for each type of cancer.Results:From 2019 to 2024,56,882 high-risk individuals were identified.The high-risk rates for lung,colorectal,breast,up-per gastrointestinal,and liver cancer were 22.02%,21.57%,14.23%,13.52%,and 6.10%,respectively.Overall,13,592 individuals com-pleted clinical screening,with detection rates of 0.32%for lung cancer,0.41%for liver cancer,0.08%for precancerous gastric lesions,3.63%for precancerous colorectal lesions,0.08%for esophageal cancer,0.16%for gastric cancer,and 0.14%for colorectal cancer.Conclusions:The implementation of the Urban Cancer Early Diagnosis and Treatment Program in Qinghai Province aids in the early detection of cancer,improves early diagnosis and survival rates,and reduces mortality.Nevertheless,due to low public awareness and limited participation,en-hancements in program management and public outreach are required.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.