1.Establishment of a porcine small intestinal epithelial cell line with IRF8 gene knockout based on AAV-SaCas9
Mingliang ZHANG ; Kaiqi LIAN ; Yao WANG ; Bingqian WANG ; Shengming MA ; Yifan ZHANG ; Xinying JI ; Xuekun DOU ; Longfei ZHANG ; Shaoting WENG
Chinese Journal of Veterinary Science 2025;45(6):1169-1177
The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P<0.05)and IFN-λ(P<0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.
2.Study on mechanical stability of maxillary LeFort Ⅰ type osteotomy after fixation with mortise and tenon and absorbable plate
Yankun ZHANG ; Zixuan SHEN ; Ran ZHANG ; Lijia DONG ; Yu TIAN ; Li LU ; Mingliang YANG ; Guangqi YAN
Chinese Journal of Stomatology 2025;60(6):635-643
Objective:To evaluate the mechanical stability of the bone block after LeFort Ⅰ osteotomy with maxillary advancement using absorbable plates fixed with tenon-and-mortise structures.Methods:This study developed three finite element models: one for the maxillary LeFort Ⅰ osteotomy with anterior advancement fixed with absorbable plates (Model 1); another for the maxillary LeFort Ⅰ osteotomy with anterior advancement fixed with absorbable plates assisted by tenon-and-mortise structures (Model 2); and the last one for the maxillary LeFort Ⅰ osteotomy with anterior advancement fixed with titanium plates and screws (Model 3). Simulated occlusal forces were applied on the anterior and posterior teeth in each model. The displacement changes of the nasal-palatine point (NP) and posterior nasal spine point (PNS) in the finite element coordinate system were compared and analyzed. The Mises equivalent stress distributions of the metal and absorbable plates were also examined to assess the mechanical stability of the three finite element models. Clinical data from 45 patients with dentofacial deformities treated from January 2017 to January 2023 at the Stomatology Hospital of China Medical University were collected. The age of the patients was 21±3 years. Among these, 15 patients had absorbable plates for fixation, 15 had absorbable plates assisted by tenon-and-mortise structures, and 15 had titanium plates and screws fixation after maxillary advancement. All patients underwent preoperative (T0), postoperative 3 days (T1), and 6 months (T2) spiral CT scans. The CT data in DICOM format were input into digital software, which was used to calculate the distances from the NP and PNS points to the horizontal plane (HP), right sagittal plane (FZSR), and coronal plane (CP) at T1 and T2. The distances at T1 and T2 were statistically analyzed using the Wilcoxon signed-rank test with SPSS 20.0, and a P value of<0.05 was considered statistically significant.Results:The finite element analysis showed that in the absorbable plate-only fixation group, the maximum displacement of the NP point (mm) under anterior and posterior tooth force conditions were 0.6 and 0.12, respectively, and for the PNS point, the maximum displacements were 0.5 and 0.11. In the tenon-and-mortise-assisted absorbable plate fixation group, the displacement of the NP point was 0.40 and 0.02 mm, and the displacement of the PNS point was 0.5 and 0.015 mm. In the titanium plate-screw fixation group, the NP point displacement was 0.33 and 0.055 mm, and the PNS point displacement was 0.16 and 0.1 mm. The Mises equivalent stress on the absorbable plates with tenon-and-mortise structure was significantly lower than that in the absorbable plate-only fixation group, while the titanium plate experienced the highest Mises equivalent stress. The clinical data analysis showed that in the horizontal direction, the postoperative stability of the three fixation methods was similar. However, in the vertical and anterior-posterior directions, the absorbable plate-only fixation group showed significant differences in the distances of PNS-HP, PNS-CP, and NP-CP between T1 and T2 ( P=0.018, P=0.009, P=0.017), suggesting significant postoperative bone displacement. In contrast, the tenon-and-mortise-assisted absorbable plate fixation group and the titanium plate-screw fixation group showed no significant differences in displacement during surgery and postoperatively(all P>0.05), demonstrating higher stability. Conclusions:The tenon-and-mortise-assisted absorbable plate fixation provides comparable stability to titanium plate fixation in clinical results, and it is more stable than absorbable plate-only fixation. In the mechanical study, when force was applied on the anterior teeth, the stability of the tenon-and-mortise-assisted absorbable plate fixation was slightly less than that of titanium plate fixation, but when posterior teeth were used, its stability exceeded both titanium plate fixation and absorbable plate-only fixation. The tenon-and-mortise-assisted absorbable plate fixation serves as an effective alternative to titanium plate fixation after LeFort Ⅰ osteotomy.
3.Influence factors for the development and regression of metabolic dysfunction-associated fatty liver disease:A study based on the health check-up population in Beijing,China
Haiqing GUO ; Xiaohui LIU ; Mingliang LI ; Feng LIU ; Yali LIU ; Jing ZHANG
Journal of Clinical Hepatology 2025;41(7):1319-1326
Objective To investigate the clinical and metabolic factors associated with the development and regression of metabolic dysfunction-associated fatty liver disease(MAFLD)in the physical examination population.Methods A retrospective observational study was conducted on 6 809 individuals who underwent physical examination in a physical examination institution in Beijing from December 2013 to December 2019,with a mean follow-up time of 52.1±13.5 months.According to the new diagnostic criteria for MAFLD,these individuals were divided into MAFLD group and non-MAFLD group,and the two groups were compared in terms of demographic indicators,body measurement indicators,and laboratory indicators at the first(baseline)and last physical examinations.The two-independent-samples t test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test was used for comparison of categorical data.A Logistic regression analysis was used to investigate the impact of various observation indicators on the development and regression of MAFLD.Results In this study,there were 4 533 individuals(66.6%)in the non-MAFLD group at baseline,among whom 15.6%developed MAFLD at the last physical examination.Compared with the non-MAFLD population,the MAFLD population had significantly higher age(Z=-6.739),number of male patients(χ2=178.534),body weight(Z=-22.302),body mass index(BMI)(Z=-22.818),waist circumference(Z=-23.117),hip circumference(Z=-18.446),systolic blood pressure(SBP)(Z=-13.301),diastolic blood pressure(DBP)(Z=-13.491),fasting blood glucose(FBG)(Z=-11.787),triglyceride(TG)(Z=-16.623),low-density lipoprotein cholesterol(LDL-C)(Z=-10.256),alanine aminotransferase(ALT)(Z=-14.250),aspartate aminotransferase(AST)(Z=-7.481),and proportion of patients with metabolic syndrome(MetS)at baseline(χ2=185.283),and there were more patients with increases in body weight,waist circumference,hip circumference,TG,TC,ALT,and AST at the final physical examination(all P<0.05);these patients had a lower level of HDL-C at baseline(Z=15.416),and there were more patients with a reduction at the last physical examination(P<0.05).There were 2 276 individuals(33.4%)in the MAFLD group at baseline,among whom 23.8%showed regression of MAFLD at the last physical examination.Compared with the population without regression of MAFLD,the population with regression of MAFLD had a significantly younger age(Z=2.185),a significantly higher number of female patients(χ2=0.340),significantly lower levels of body weight(Z=-8.909),BMI(Z=-10.205),waist circumference(Z=-11.183),hip circumference(Z=-7.178),SBP(Z=-3.627),DBP(Z=-3.443),TG(Z=-5.945),ALT(Z=-9.664),and AST(Z=-5.904),and a significantly lower proportion of patients with MetS(χ2=42.082),and there were more patients with reductions in body weight,waist circumference,hip circumference,blood pressure,TG,TC,ALT,and AST at the final physical examination(all P<0.05);these patients had a higher level of HDL-C at baseline(Z=6.778),and there were more patients with an increase at the last physical examination(P<0.05).The multivariate Logistic regression analysis showed that sex and changes in body weight and HDL-C during physical examination were independently associated with the development and regression of MAFLD(all P<0.05).Conclusion There is a relatively high prevalence rate of MAFLD among the physical examination population in Beijing,with a higher proportion of male patients.There are significant metabolic disorders and liver function abnormalities,and changes in body weight and HDL-C are the most important predictive indicators for the development and regression of MAFLD.
4.The application of surgical robots in head and neck tumors.
Xiaoming HUANG ; Qingqing HE ; Dan WANG ; Jiqi YAN ; Yu WANG ; Xuekui LIU ; Chuanming ZHENG ; Yan XU ; Yanxia BAI ; Chao LI ; Ronghao SUN ; Xudong WANG ; Mingliang XIANG ; Yan WANG ; Xiang LU ; Lei TAO ; Ming SONG ; Qinlong LIANG ; Xiaomeng ZHANG ; Yuan HU ; Renhui CHEN ; Zhaohui LIU ; Faya LIANG ; Ping HAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(11):1001-1008
5.HOTAIR rs920778 single nucleotide polymorphism is associated with breast cancer susceptibility and HER2-targeted therapy resistance in Chinese population.
Mingliang ZHANG ; Feifan SUN ; Zhuoqi HAN ; Yue GAO ; Yi LUO
Journal of Southern Medical University 2025;45(10):2270-2276
OBJECTIVES:
To investigate the association of HOTAIR gene rs920778 single nucleotide polymorphism (SNP) with breast cancer susceptibility and response to HER2-targeted therapy in a Chinese population.
METHODS:
TaqMan probe-based real-time quantitative PCR was used for genotyping of the rs920778 locus (chr12:54,376,218) in peripheral blood genomic DNA from 287 breast cancer patients and 260 healthy individuals from northern Anhui Province. The genotype (GG, GT and TT) and allele (G/T) distribution frequencies were compared between the two groups to evaluate their association with breast cancer risk. Multivariate logistic regression analysis was conducted to assess the relationship between SNP at this locus and aggressive clinicopathological features (including tumor size, lymph node metastasis, ER/PR/HER2 status, and molecular subtypes) of breast cancer. For the HER2-positive subgroup, the association between rs920778 genotype and responses to dual-targeted therapy (trastuzumab [6 mg/kg q3w]+pertuzumab [420 mg q3w] + docetaxel [75 mg/m²]) was analyzed. The primary endpoints included pathological complete response rate (pCR), objective response rate (ORR), and progression-free survival (PFS).
RESULTS:
The TT genotype of rs920778 was associated with a significantly increased breast cancer susceptibility (OR=1.54, 95% CI: 1.09-2.19; P=0.017), an advanced tumor stage (P<0.001), lymph node metastasis (P<0.001), and the triple-negative subtype (P<0.001). In HER2-positive patients, TT genotype carriers had a markedly reduced objective response rate to dual HER2-targeted therapy (33.3% vs 89.3%, P=0.001) and a lower pathological complete response rate after neoadjuvant therapy (P=0.018).
CONCLUSIONS
The TT genotype of HOTAIR rs920778 serves as an independent risk factor for breast cancer susceptibility and aggressive progression in Chinese population and may predict the resistance to HER2-targeted therapies, suggesting its potential as a prognostic biomarker for precision oncology.
Adult
;
Aged
;
Female
;
Humans
;
Middle Aged
;
Breast Neoplasms/drug therapy*
;
Case-Control Studies
;
China
;
Drug Resistance, Neoplasm/genetics*
;
Genetic Predisposition to Disease
;
Genotype
;
Polymorphism, Single Nucleotide
;
Receptor, ErbB-2
;
RNA, Long Noncoding/genetics*
;
East Asian People/genetics*
6.Analysis of association of IL-23R gene polymorphisms with susceptibility for psoriasis.
Quan GAN ; Lixia WANG ; Beibei WANG ; Manman ZHANG ; Mingliang DONG ; Beibei SU
Chinese Journal of Medical Genetics 2025;42(4):505-511
OBJECTIVE:
To assess the association of single nucleotide polymorphisms (SNPs) of interleukin-23 receptor (IL-23R) gene with susceptibility to psoriasis.
METHODS:
Two hundred and ten psoriasis patients admitted to Xinxiang Central Hospital from January 2019 to December 2024 were selected as the study group, and 210 healthy individuals undergoing physical examination during the same period were selected as the control group. 3 mL of peripheral venous blood sample was collected from each individual from the two groups, and PCR-Restriction fragment length polymorphism (PCR-RFLP) assay was used to determine the polymorphisms of the IL-23R gene at rs2201841, rs1004819, rs10889677, rs1343151 and rs1495965 loci. Genotypic and allelic distribution of each SNP locus was calculated to assess the association between SNPs of the IL-23R gene with the onset of psoriasis, and the difference in serum IL-23 levels among patients with different genotypes at each locus was compared. This study was approved by the Medical Ethics Committee of Xinxiang Central Hospital (Ethic No. 2024-749).
RESULTS:
The results showed that the frequency of CC genotype at rs1004819 locus of the study group was significantly higher than that of the control group (26.19% vs. 18.10%, P < 0.05), and the frequency of C allele was also significantly higher than that of the control group (54.05% vs. 42.62%, P < 0.05). There was no significant difference in allelic and genotypic frequencies between the two groups at rs2201841, rs10889677, rs1343151, and rs1495965 loci (P > 0.05). The dominant and recessive inheritance patterns at the rs1004819 locus are associated with susceptibility to psoriasis (P < 0.05), while the different inheritance patterns at rs2201841, rs10889677, rs1343151, and rs1495965 loci are not associated with psoriasis (P > 0.05). The serum IL-23 levels of patients with CC genotype at the rs1004819 locus were higher than those with the CT and TT genotypes (P < 0.05). No significant difference was detected in the serum levels of IL-23 between patients with different genotypes for the rs2201841, rs10889677, rs1343151, and rs1495965 loci (P > 0.05).
CONCLUSION
The polymorphism at the rs1004819 locus of the IL-23R gene is associated with susceptibility to psoriasis, and individuals carrying the CC genotype and C allele have a higher risk of developing the disease.
Humans
;
Psoriasis/genetics*
;
Polymorphism, Single Nucleotide
;
Receptors, Interleukin/genetics*
;
Genetic Predisposition to Disease
;
Male
;
Female
;
Adult
;
Middle Aged
;
Genotype
;
Alleles
;
Gene Frequency
;
Case-Control Studies
;
Interleukin-23/blood*
7.Prevalence, influencing factors, and fibrosis risk stratification of metabolic dysfunction-associated fatty liver disease in the health check-up population in Beijing, China
Haiqing GUO ; Mingliang LI ; Feng LIU ; Jing ZHANG
Journal of Clinical Hepatology 2025;41(4):643-649
ObjectiveTo identify the patients with metabolic dysfunction-associated fatty liver disease (MAFLD) among the health check-up population, and to perform stratified management of patients with the low, medium, and high risk of advanced fibrosis based on noninvasive fibrosis scores. MethodsA cross-sectional study was conducted among 3 125 individuals who underwent physical examination in Beijing Physical Examination Center from December 2017 to December 2019, and they were divided into MAFLD group with 1 068 individuals and non-MAFLD group with 2 057 individuals. According to BMI, the MAFLD group was further divided into lean MAFLD group (125 individuals with BMI<24 kg/m2) and non-lean MAFLD group (943 individuals with BMI≥24 kg/m2). Indicators including demographic data, past history, laboratory examination, and liver ultrasound were compared between groups. Fibrosis-4 (FIB-4) score, NAFLD fibrosis score (NFS), aspartate aminotransferase-to-platelet ratio index (APRI), and BARD score were calculated for the patients in the MAFLD group to assess the risk of advanced fibrosis. The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. A logistic regression analysis was used to investigate the influence of each indicator in MAFLD. ResultsCompared with the non-MAFLD group, the MAFLD group had significantly higher age (Z=-9.758, P<0.05), proportion of male patients (χ2=137.555, P<0.05), and levels of body weight (Z=-27.987, P<0.05), BMI (Z=-32.714, P<0.05), waist circumference (Z=-31.805, P<0.05), hip circumference (Z=-26.342, P<0.05), waist-hip ratio (Z=-28.554, P<0.05), alanine aminotransferase (ALT) (Z=-25.820, P<0.05), aspartate aminotransferase (AST) (Z=-16.894, P<0.05), gamma-glutamyl transpeptidase (GGT) (Z=-25.069, P<0.05), alkaline phosphatase (Z=-12.533, P<0.05), triglyceride (Z=-27.559), total cholesterol (Z=-7.833, P<0.05), low-density lipoprotein cholesterol (LDL-C) (Z=-8.222, P<0.05), and uric acid (UA) (Z=-20.024, P<0.05), as well as a significantly higher proportion of patients with metabolic syndrome (MetS) (χ2=578.220, P<0.05), significantly higher prevalence rates of hypertension (χ2=241.694, P<0.05), type 2 diabetes (χ2=796.484, P<0.05), and dyslipidemia (χ2=369.843, P<0.05), and a significant reduction in high-density lipoprotein cholesterol (HDL-C) (Z=23.153, P<0.001). The multivariate logistic regression analysis showed that male sex (odds ratio [OR]=1.45, 95% confidence interval [CI]: 1.203 — 1.737), ALT (OR=1.05, 95%CI: 1.046 — 1.062), LDL-C (OR=1.23, 95%CI: 1.102 — 1.373), and comorbidity with MetS (OR=5.97, 95%CI: 4.876 — 7.316) were independently associated with MAFLD. Compared with the non-lean MAFLD group, the lean MAFLD group had significantly higher age (Z=3.736, P<0.05) and HDL-C (Z=2.679, P<0.05) and significant reductions in the proportion of male patients (χ2=28.970, P<0.05), body weight (Z=-14.230, P<0.05), BMI (Z=-18.188, P<0.05), waist circumference (Z=-13.451, P<0.05), hip circumference (Z=-13.317, P<0.05), ALT (Z=-4.519, P<0.05), AST (Z=-2.258, P<0.05), GGT (Z=-4.592, P<0.05), UA (Z=-4.415, P<0.05), the proportion of patients with moderate or severe fatty liver disease or MetS (χ2=42.564, P<0.05), and the prevalence rates of hypertension (χ2=12.057, P<0.05) and type 2 diabetes (χ2=3.174, P<0.05). Among the patients with MAFLD, 10 patients (0.9%) had an FIB-4 score of >2.67, 4 patients (0.4%) had an NFS score of >0.676, 8 patients (0.7%) had an APRI of >1, and 551 patients (51.6%) had a BARD score of ≥2. ConclusionThere is a relatively high prevalence rate of MAFLD among the health check-up population in Beijing, but with a relatively low number of patients with a high risk of advanced fibrosis, and such patients need to be referred to specialized hospitals for liver diseases.
8.Analysis of psychological crisis related factors of college students based on the dual factor model of mental health
SUN Yujing, YIN Fei, WANG Mingliang, JIANG Wenlong, ZHANG Jing
Chinese Journal of School Health 2025;46(6):847-851
Objective:
To analyze the current status and influencing factors of psychological crisis among college students, so as to provide a scientific basis for the formulation of psychological crisis intervention plans in colleges and universities.
Methods:
From September to December 2024, 645 college students from a medical undergraduate university in Heilongjiang Province were selected with a convenience sampling method. A convergent mixed analysis design was used. Quantitative analysis was conducted using College Students Psychological Crisis Screening Scale, Emotion Regulation Questionnaire, Short-Egna Minnen av Barndoms Uppfostran and Perceived School Climate Scale. Binary Logistic regression analysis was used to explore the related factors of psychological crisis among college students. Qualitative research was conducted on 15 college students with psychological crisis identified in the quantitative analysis by a purposive sampling method. The interview data were organized and analyzed using the thematic framework analysis method.
Results:
Among the surveyed college students, 92 (14.3%) had psychological crisis. Binary Logistic regression analysis results showed that positive parenting style ( OR=0.97,95%CI =0.95-0.99), negative parenting style ( OR=1.01,95%CI =1.00-1.02), cognitive reappraisal ( OR=0.88, 95%CI =0.83-0.92), expressive suppression ( OR=1.08, 95%CI =1.02-1.15), and perceived campus atmosphere ( OR=0.97, 95%CI =0.95-0.98) were all related factors of psychological crisis among college students ( P <0.05). The qualitative analysis results showed that there were three themes for the influencing factors of college students psychological crisis, including differential impact of emotion regulation strategies on psychological state, shaping of psychological state of college students by family and bidirectional effect of perceived campus atmosphere on psychological state. Mixed analysis results showed that the influencing factors of college students psychological crisis were consistent in terms of emotion regulation strategies, and were expansive in terms of parenting style and perceived campus atmosphere.
Conclusion
Schools and mental health service departments can reduce the risk of psychological crisis by optimizing cognitive reappraisal and reducing expressive suppression, improve the level of psychological crisis by strengthening positive family interaction and blocking negative parenting style, and maintain the mental health level of college students by building a supportive campus environment and alleviating high pressure.
9.Characteristics and lifestyles of patients with metabolic dysfunction-associated fatty liver disease based on the physical examination population
Haiqing GUO ; Mingliang LI ; Feng LIU ; Yali LIU ; Jing ZHANG
Journal of Clinical Hepatology 2025;41(6):1090-1096
ObjectiveTo screen for the patients with metabolic dysfunction-associated fatty liver disease (MAFLD) among the physical examination population, to observe the characteristics of MAFLD patients, and to compare the differences in lifestyle between the MAFLD population and the non-MAFLD population. MethodsA cross-sectional study was conducted among 6 206 individuals who underwent physical examination in a physical examination institution in Beijing from December 2015 to December 2019, and according to the new diagnostic criteria for MAFLD, the examination population was divided into MAFLD group and non-MAFLD group. Based on body mass index (BMI), the MAFLD group was further divided into lean MAFLD group (BMI<24 kg/m2) and non-lean MAFLD group (BMI ≥24 kg/m2). Related data were compared between groups, including demographic indicators, education level, work pressure, physical measurement indicators, and lifestyles such as sleep, diet, and exercise. The independent-samples t test was used for comparison of normally distributed continuous data between two groups, the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, and the chi-square test was used for comparison of categorical data between groups. ResultsOf all individuals in this study, 1 926 (31.1%) had MAFLD and 4 280 (68.9%) did not have MAFLD. Compared with the non-MAFLD group, the MAFLD group had significantly higher age (Z=-14.459, P<0.001), proportion of male patients (χ2=72.004, P<0.001), work pressure (χ2=7.744, P=0.005), body weight (Z=-43.508, P<0.001), BMI (Z=-47.621, P<0.001), waist circumference (Z=-48.515, P<0.001), hip circumference (Z=-42.121, P<0.001), and waist-hip ratio (Z=-43.535, P<0.001), as well as a significantly lower education level (χ2=33.583, P<0.001). In terms of behavior, the MAFLD group had a significantly shorter sleep time (χ2=5.820, P=0.016) and a significantly faster eating speed (χ2=74.476, P<0.001). In terms of diet, the patients in the MAFLD group consumed more high-sodium, high-sugar, and high-calorie diets (χ2=42.667, P<0.001) and low-fiber diet (χ2=4.367, P=0.008). In terms of exercise, the MAFLD group had a significantly higher proportion of patients without exercise habits (χ2=10.278, P=0.001). Further analysis showed that there were 202 individuals (10.5%) in the lean MAFLD group and 1 724 (89.5%) in the non-lean MAFLD group. Compared with the non-lean MAFLD group, the lean MAFLD group had significantly higher age (Z=3.368, P=0.001) and education level (χ2=9.647, P=0.002) and significantly lower proportion of male patients (χ2=27.664, P<0.001), body weight (Z=-18.483, P<0.001), BMI (Z=-23.286, P<0.001), waist circumference (Z=-18.565, P<0.001), and hip circumference (Z=-18.097, P<0.001), and in terms of behavior, the non-lean MAFLD group had a significantly faster eating speed (χ2=4.549, P=0.033). ConclusionThere is a relatively high prevalence rate of MAFLD among the physical examination population in Beijing, with a higher number of people with unhealthy lifestyles compared with the non-MAFLD population.
10.Establishment of a porcine small intestinal epithelial cell line with IRF8 gene knockout based on AAV-SaCas9
Mingliang ZHANG ; Kaiqi LIAN ; Yao WANG ; Bingqian WANG ; Shengming MA ; Yifan ZHANG ; Xinying JI ; Xuekun DOU ; Longfei ZHANG ; Shaoting WENG
Chinese Journal of Veterinary Science 2025;45(6):1169-1177
The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P<0.05)and IFN-λ(P<0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.


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