Objective:To improve the isolation and culture method of seasonal influenza virus in embryonated chicken eggs (ECEs), and evaluate their isolation efficiency.Methods:We randomly selected 80 positive samples of H1N1 (H1N1pdm09) and seasonal H3N2 (H3N2snl) influenza virus nucleic acid, and inoculated them into the amniotic and urinary sac cavities of 10-day-old (traditional method) and 14-day-old (improved method) ECEs respectively to adapt the virus to the ECEs (E1-E2). Both method were used to inoculate 10-day-old urinary sac amplification virus (E2-E3), and the final virus isolation positive rates of the two method were compared; using fluorescence quantitative PCR method to detect viral nucleic acids in the improved amniotic and urinary sac cultures, and evaluate the viral proliferation at different inoculation sites; we analyzed the correlation between virus content and isolation positivity rate in the original specimen based on the CT value of nucleic acid testing and the final virus isolation positivity rate using the improved method.Results:The improved method obtained 42 strains of H1N1pdm09 strain, with a positive rate of 52.5% ( χ2=38.571, P<0.01); obtained 54 strains of H3N2snl strain, with a positive rate of 67.5% ( χ2=40.921, P<0.01). Significant differences were observed in the isolation efficiency of H1N1pdm09 samples when the improved method was applied to different inoculation sites of chicken embryos ( χ2=30.476, P<0.01), and similar differences were noted for H3N2snl samples ( χ2=4.928, P=0.026). There was no significant difference in the isolation rate of different CT value intervals of the original samples ( χH1N1pdm092=10.226, χH3N2snl2=3.764, P>0.05). Conclusions:The improved method of inoculating 14-day old ECEs adapted the virus, and the final number of strains obtained was significantly higher than the traditional method of inoculating 10 day old ECEs, which can significantly improve the positive isolation rate of H1N1pdm09 and H3N2snl influenza virus in ECEs. The amniotic cavity is more sensitive to H1N1pdm09 and H3N2snl influenza viruses, which helps the virus adapt in ECEs. There was no significant difference in the sample isolation rate and total positive rate of virus isolation among different CT value ranges, and further verification is needed.

Objective To explore the risk factors of nonalcoholic fatty liver disease(NAFLD)in obese children,and evaluate the diagnostic value of each index for NAFLD and establish a Nomogram prediction model.Methods A total of 207 obese children admitted at Department of Pediatrics the Second Hospital of Jiaxing from January 2022 to January 2025 were selected.These children were divided into two groups based on NAFLD diagnosis:non-NAFLD group(n=99)and NAFLD group(n=108).Differences in gender,age,body mass index(BMI),and related metabolic indicators were compared between two groups.Logistic regression was employed to analyze potential risk factors for NAFLD development,while receiver operating characteristic(ROC)curve and Nomograms were used to evaluate the predictive value of different factors for NAFLD.Results ROC curve analysis demonstrated diagnostic value for NAFLD in triglyceride-glucose index(TyG),triglyceride-to-cholesterol ratio,TyG-waist circumference,and TyG-BMI.Among these,the area under the curve(AUC)of TyG showed the highest value of 0.713,with an optimal cutoff of 8.189,sensitivity of 50.5％and specificity of 83.3％.Univariate Logistic regression analysis revealed multiple insulin resistance indicators associated with NAFLD development.Multivariate analysis identified homeostatic model assessment of insulin resistance(HOMA-IR)and TyG as independent risk factors,with TyG showing the best predictive value(OR=3.038 95％CI:1.089-8.475,P＜0.05).The constructed Nomogram prediction model demonstrated strong comprehensive discriminant capability(AUC=0.742).Conclusion The Nomogram model based on HOMA-IR,TyG and its derived indexes has certain clinical application value in the screening of NAFLD in children.

Objective To screen immune-related long non-coding RNAs(LncRNAs)in the TCGA database pancreatic cancer dataset and construct a prognostic risk assessment model with immune-related LncRNAs to explore prognosis-related potential molecular mechanisms.Methods RNA-seq data of 171 pancreatic cancer samples and corresponding clinical information were obtained by The Cancer Genome Atlas(TCGA)database,and two classical immune-related gene datasets(GO0006955/IMMUNE RESPONSE and GO0002376/IMMUNE SYSTERM PROCESS)and gene annotation information were used to identify immune-related LncRNAs.The immune-related LncRNAs associated with pancreatic cancer prognosis were used for univariate and multivariate Cox analyses to establish a model for the assessment of pancreatic cancer prognostic risk based on immune-associated LncRNAs.This risk model was used for survival analysis,clinical correlation analysis,immune cell infiltration analysis,pathway enrichment analysis,and prognostic column line plot modeling.Results We screened 119 immune-related LncRNAs in pancreatic cancer,and five immune-related LncRNAs(AC064836.3,LINC00941,ZNF236-DT,TMEM161B-AS1 and AC068580.2)were identified for the development of pancreatic cancer prognostic risk assessment model.According to the prognostic risk assessment model,pancreatic cancer patients were divided into low-risk group(n=86)and high-risk group(n=85).Compared with the low-risk group,the high-risk group showed a significant negative enrichment trend for immune-related signaling pathways,the 5-year overall survival of pancreatic cancer patients was significantly increased in the low-risk group compared with the high-risk group.The expression of low-risk immune-related LncRNAs(AC064836.3,ZNF236-DT and TMEM161B-AS1)gradually decreased with increasing clinical stage of pancreatic cancer patients.Patient age(P=0.031,risk ratio and 95％CI:1.025/1.002-1.048)and prognostic risk score(P＜0.001,risk ratio and 95％ confidence interval 1.801/1.465-2.215)could be used as independent prognostic risk factors for overall survival in pancreatic cancer.In addition,the prognostic risk assessment model had better predictive efficiency(area under the curve=0.695)compared with the disease predictive ability of common clinical characteristics.Steroid biosynthesis,pentose phosphate pathway,intercellular linkage,cytoskeletal rearrangement and other pathways related to energy metabolism and invasive migration of pancreatic cancer cells were significantly activated in the high-risk group.Meanwhile,pancreatic cancer patients in the high-risk group had lower levels of naive B cells,plasma cells and neutrophils with anti-tumor activity,but their macrophage infiltration levels were significantly higher than those in the low-risk group.Conclusion The prognostic risk assessment model constructed based on five immune-related LncRNAs can effectively predict the survival status,clinical characteristics,molecular pathways,and immune cell infiltration differences of pancreatic cancer patients.Meanwhile,relying on this model,the prognosis of pancreatic cancer patients can be prospectively predicted,which enhances the usefulness of this risk prediction model.

Objective:To explore the effect of contezolid on platelet count in patients with severe pneumonia and analyze the risk factors.Methods:The study was designed as a retrospective case-control study. The research subjects were selected from patients with severe pneumonia who were admitted to the Department of Respiratory and Critical Care Medicine of Shaanxi Provincial People′s Hospital from July 1, 2022 to November 30, 2024 and were treated with contezolid or linezolid. The clinical data of patients were collected and the incidence of thrombocytopenia [platelet count (PLT)<100×10 9/L after medication], the PLT before and at 1 week of treatments, and the lowest PLT value during treatments were compared in patients treated with contezolid and linezolid. The patients were divided into 2 groups based on whether contezolid- related thrombocytopenia occurred. The clinical characteristics of the patients were compared, and the independent risk factors of contezolid-related thrombocytopenia were analyzed by binary logistic regression method. Results:A total of 175 patients were included, among whom 73 received contezolid and 102 received linezolid. There was no statistically significant difference in PLT between the 2 groups before medication ( P=0.364). Compared with patients treated with linezolid, the incidence of thrombocytopenia in patients treated with contezolid was lower [19.2% (14/73) vs. 40.2% (41/102)], and the PLT at 1 week of treatments and the lowest value of PLT during treatments were higher. The differences were all statistically significant (all P<0.05). Compared with patients without contezolid-related thrombocytopenia(59 patients), patients who developed thrombocytopenia after using contezolid had a longer duration of contezolid medication, lower PLT and creatinine clearance rate before medication, and higher procalcitonin and serum creatinine levels before medication. All these differences were statistically significant (all P<0.05). The results of binary logistic regression analysis showed that lower PLT [odds ratio ( OR)=0.971, 95% confidence interval ( CI): 0.950-0.992, P=0.008] and higher procalcitonin level ( OR=7.292, 95% CI: 1.067-49.814, P=0.043) before medication and longer duration of contezolid medication ( OR=1.165, 95% CI: 1.002-1.355, P=0.046) were the independent risk factors of contezolid-related thrombocytopenia. Conclusions:Compared with linezolid, contezolid has a relatively safer profile in the treatment of patients with severe pneumonia and the risk of thrombocytopenia after medication is lower. Patients with lower PLT and higher procalcitonin levels before medication, and those with a longer duration of contezolid medication have a higher risk of contezolid-related thrombocytopenia and should be closely monitored.

Objective To screen immune-related long non-coding RNAs(LncRNAs)in the TCGA database pancreatic cancer dataset and construct a prognostic risk assessment model with immune-related LncRNAs to explore prognosis-related potential molecular mechanisms.Methods RNA-seq data of 171 pancreatic cancer samples and corresponding clinical information were obtained by The Cancer Genome Atlas(TCGA)database,and two classical immune-related gene datasets(GO0006955/IMMUNE RESPONSE and GO0002376/IMMUNE SYSTERM PROCESS)and gene annotation information were used to identify immune-related LncRNAs.The immune-related LncRNAs associated with pancreatic cancer prognosis were used for univariate and multivariate Cox analyses to establish a model for the assessment of pancreatic cancer prognostic risk based on immune-associated LncRNAs.This risk model was used for survival analysis,clinical correlation analysis,immune cell infiltration analysis,pathway enrichment analysis,and prognostic column line plot modeling.Results We screened 119 immune-related LncRNAs in pancreatic cancer,and five immune-related LncRNAs(AC064836.3,LINC00941,ZNF236-DT,TMEM161B-AS1 and AC068580.2)were identified for the development of pancreatic cancer prognostic risk assessment model.According to the prognostic risk assessment model,pancreatic cancer patients were divided into low-risk group(n=86)and high-risk group(n=85).Compared with the low-risk group,the high-risk group showed a significant negative enrichment trend for immune-related signaling pathways,the 5-year overall survival of pancreatic cancer patients was significantly increased in the low-risk group compared with the high-risk group.The expression of low-risk immune-related LncRNAs(AC064836.3,ZNF236-DT and TMEM161B-AS1)gradually decreased with increasing clinical stage of pancreatic cancer patients.Patient age(P=0.031,risk ratio and 95％CI:1.025/1.002-1.048)and prognostic risk score(P＜0.001,risk ratio and 95％ confidence interval 1.801/1.465-2.215)could be used as independent prognostic risk factors for overall survival in pancreatic cancer.In addition,the prognostic risk assessment model had better predictive efficiency(area under the curve=0.695)compared with the disease predictive ability of common clinical characteristics.Steroid biosynthesis,pentose phosphate pathway,intercellular linkage,cytoskeletal rearrangement and other pathways related to energy metabolism and invasive migration of pancreatic cancer cells were significantly activated in the high-risk group.Meanwhile,pancreatic cancer patients in the high-risk group had lower levels of naive B cells,plasma cells and neutrophils with anti-tumor activity,but their macrophage infiltration levels were significantly higher than those in the low-risk group.Conclusion The prognostic risk assessment model constructed based on five immune-related LncRNAs can effectively predict the survival status,clinical characteristics,molecular pathways,and immune cell infiltration differences of pancreatic cancer patients.Meanwhile,relying on this model,the prognosis of pancreatic cancer patients can be prospectively predicted,which enhances the usefulness of this risk prediction model.

Objective To explore the risk factors of nonalcoholic fatty liver disease(NAFLD)in obese children,and evaluate the diagnostic value of each index for NAFLD and establish a Nomogram prediction model.Methods A total of 207 obese children admitted at Department of Pediatrics the Second Hospital of Jiaxing from January 2022 to January 2025 were selected.These children were divided into two groups based on NAFLD diagnosis:non-NAFLD group(n=99)and NAFLD group(n=108).Differences in gender,age,body mass index(BMI),and related metabolic indicators were compared between two groups.Logistic regression was employed to analyze potential risk factors for NAFLD development,while receiver operating characteristic(ROC)curve and Nomograms were used to evaluate the predictive value of different factors for NAFLD.Results ROC curve analysis demonstrated diagnostic value for NAFLD in triglyceride-glucose index(TyG),triglyceride-to-cholesterol ratio,TyG-waist circumference,and TyG-BMI.Among these,the area under the curve(AUC)of TyG showed the highest value of 0.713,with an optimal cutoff of 8.189,sensitivity of 50.5％and specificity of 83.3％.Univariate Logistic regression analysis revealed multiple insulin resistance indicators associated with NAFLD development.Multivariate analysis identified homeostatic model assessment of insulin resistance(HOMA-IR)and TyG as independent risk factors,with TyG showing the best predictive value(OR=3.038 95％CI:1.089-8.475,P＜0.05).The constructed Nomogram prediction model demonstrated strong comprehensive discriminant capability(AUC=0.742).Conclusion The Nomogram model based on HOMA-IR,TyG and its derived indexes has certain clinical application value in the screening of NAFLD in children.

Objective:To explore the effect of contezolid on platelet count in patients with severe pneumonia and analyze the risk factors.Methods:The study was designed as a retrospective case-control study. The research subjects were selected from patients with severe pneumonia who were admitted to the Department of Respiratory and Critical Care Medicine of Shaanxi Provincial People′s Hospital from July 1, 2022 to November 30, 2024 and were treated with contezolid or linezolid. The clinical data of patients were collected and the incidence of thrombocytopenia [platelet count (PLT)<100×10 9/L after medication], the PLT before and at 1 week of treatments, and the lowest PLT value during treatments were compared in patients treated with contezolid and linezolid. The patients were divided into 2 groups based on whether contezolid- related thrombocytopenia occurred. The clinical characteristics of the patients were compared, and the independent risk factors of contezolid-related thrombocytopenia were analyzed by binary logistic regression method. Results:A total of 175 patients were included, among whom 73 received contezolid and 102 received linezolid. There was no statistically significant difference in PLT between the 2 groups before medication ( P=0.364). Compared with patients treated with linezolid, the incidence of thrombocytopenia in patients treated with contezolid was lower [19.2% (14/73) vs. 40.2% (41/102)], and the PLT at 1 week of treatments and the lowest value of PLT during treatments were higher. The differences were all statistically significant (all P<0.05). Compared with patients without contezolid-related thrombocytopenia(59 patients), patients who developed thrombocytopenia after using contezolid had a longer duration of contezolid medication, lower PLT and creatinine clearance rate before medication, and higher procalcitonin and serum creatinine levels before medication. All these differences were statistically significant (all P<0.05). The results of binary logistic regression analysis showed that lower PLT [odds ratio ( OR)=0.971, 95% confidence interval ( CI): 0.950-0.992, P=0.008] and higher procalcitonin level ( OR=7.292, 95% CI: 1.067-49.814, P=0.043) before medication and longer duration of contezolid medication ( OR=1.165, 95% CI: 1.002-1.355, P=0.046) were the independent risk factors of contezolid-related thrombocytopenia. Conclusions:Compared with linezolid, contezolid has a relatively safer profile in the treatment of patients with severe pneumonia and the risk of thrombocytopenia after medication is lower. Patients with lower PLT and higher procalcitonin levels before medication, and those with a longer duration of contezolid medication have a higher risk of contezolid-related thrombocytopenia and should be closely monitored.

Objective:To improve the isolation and culture method of seasonal influenza virus in embryonated chicken eggs (ECEs), and evaluate their isolation efficiency.Methods:We randomly selected 80 positive samples of H1N1 (H1N1pdm09) and seasonal H3N2 (H3N2snl) influenza virus nucleic acid, and inoculated them into the amniotic and urinary sac cavities of 10-day-old (traditional method) and 14-day-old (improved method) ECEs respectively to adapt the virus to the ECEs (E1-E2). Both method were used to inoculate 10-day-old urinary sac amplification virus (E2-E3), and the final virus isolation positive rates of the two method were compared; using fluorescence quantitative PCR method to detect viral nucleic acids in the improved amniotic and urinary sac cultures, and evaluate the viral proliferation at different inoculation sites; we analyzed the correlation between virus content and isolation positivity rate in the original specimen based on the CT value of nucleic acid testing and the final virus isolation positivity rate using the improved method.Results:The improved method obtained 42 strains of H1N1pdm09 strain, with a positive rate of 52.5% ( χ2=38.571, P<0.01); obtained 54 strains of H3N2snl strain, with a positive rate of 67.5% ( χ2=40.921, P<0.01). Significant differences were observed in the isolation efficiency of H1N1pdm09 samples when the improved method was applied to different inoculation sites of chicken embryos ( χ2=30.476, P<0.01), and similar differences were noted for H3N2snl samples ( χ2=4.928, P=0.026). There was no significant difference in the isolation rate of different CT value intervals of the original samples ( χH1N1pdm092=10.226, χH3N2snl2=3.764, P>0.05). Conclusions:The improved method of inoculating 14-day old ECEs adapted the virus, and the final number of strains obtained was significantly higher than the traditional method of inoculating 10 day old ECEs, which can significantly improve the positive isolation rate of H1N1pdm09 and H3N2snl influenza virus in ECEs. The amniotic cavity is more sensitive to H1N1pdm09 and H3N2snl influenza viruses, which helps the virus adapt in ECEs. There was no significant difference in the sample isolation rate and total positive rate of virus isolation among different CT value ranges, and further verification is needed.

Objective To investigate the mechanism of fibroblast growth factor(FGF)21 in liver lipid deposition.Methods HepG2 cells were divided into control group and induction group.Fatty liver cell model was constructed using oleic acid and palmitic acid in induction group.Lipid deposition under the microscope was observed between two groups by oil red O staining,and the protein expression of FGF21 and glutathione peroxidase 4(GPX4)in two groups were detected.FGF21 lentivirus-infected fatty liver cells were used as interference group,and scramble lentivirus-infected fatty liver cells were used as non-interference group.The expression level of FGF21 mRNA,lipid deposition and expression level of GPX4 protein were compared between two groups.Results Compared with control group,induction group had significant lipid deposition,significantly increased expression level of FGF21 protein,and significantly decreased expression level of GPX4 protein(P＜0.05).After the interference of FGF21 lentivirus,expression level of FGF21 mRNA was significantly decreased,lipid droplets were decreased,lipid deposition was significantly improved,and GPX4 protein expression level was significantly increased in interference group(P＜0.05).Conclusion FGF21 may promote liver lipid formation by inhibiting lipid peroxidation.

Objective:To investigate the clinicopathological and genetic features of confined placental mosaicism (CPM) and its effect on fetal intrauterine growth.Methods:Fourteen CPM cases of Haidian Maternal and Children Health Hospital were collected from May 2018 to March 2022. Clinicopathological examination on placental specimens and molecular genetic analysis were performed.Results:The age of the parturient women ranged from 27 to 34 years, with an average age of (30.0±3.54) years. The gestational weeks ranged from 35 +1 to 41 +2 weeks. There were 4 premature births and 10 term births, among which 6 were female and 8 were male fetuses. Nine cases (9/14) had adverse pregnancy outcomes, including 7 cases of fetal growth restriction. The weight of CPM placenta decreased, with 6 cases below the 10th percentile of weight standards and 5 cases between the 10th and 25th percentile. All 14 CPM placental specimens showed morphological changes of perfusion dysfunction to varying degrees, with mainly placental-maternal vascular malperfusion followed by placental-fetal vascular malperfusion. The mosaic chromosomes in different CPM cases varied, with 16-trisomy/monosomy mosaicism being the most common followed by 7-trisomy and 21-trisomy/monosomy mosaicism. The mosaic proportion was unequal in different parts of the same CPM placenta, with the mosaic proportion of umbilical cord, fetal membranes, fetal surface, maternal surface, and edge ranging from 1% to 70%. Conclusions:The mosaic chromosomes in different CPM cases vary, and the mosaic proportion is unequal in different parts of the same CPM placenta. The pathological morphology is mainly manifested as perfusion dysfunction, which can lead to adverse pregnancy outcomes such as fetal growth restriction and preterm birth.