Objective:To identify and validate the key genes of ferroptosis in phospholipase A2 receptor (PLA2R) associated membranous nephropathy through bioinformatics analysis and in vitro experiments, and to explore the potential role of ferroptosis in PLA2R associated membranous nephropathy (PMN). Methods:The GSE115857 dataset obtained by retrieving the Gene Expression Omnibus (GEO) database and the ferroptosis-related genes obtained by retrieving the FerrDb database were intersected. The intersected genes were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key ferroptosis genes associated with PMN were identified by intersecting genes selected using support vector machines-recursive feature elimination and least absolute shrinkage and selection operator regression. The results were validate by real-time PCR, cell counting kit-8, Western blotting and immunofluorescence in human renal podocyte line AB 8/13 from both the control group and model group.Results:A total of 25 genes related to ferroptosis of PMN were obtained, and GO and KEGG analysis showed that these genes were mainly involved in cell ferroptosis metabolism. The key ferroptosis genes of PMN obtained by machine learning method were activating transcription factor 3 ( ATF3) and coiled coil domain containing 6 ( CCDC6). The results of in vitro experiments showed that the human renal podocyte line AB 8/13 in the model group was significantly deformed and retracted compared with the control group. The surface area density of foot processes was significantly reduced, and the podocyte cytoskeleton was allosteric. The morphology of F-actin was disordered and the expression of synaptopodin was decreased. The cell proliferation activity was significantly decreased ( P<0.05). The expression of PLA2R protein was increased ( P<0.05), and the expression of GPX4 protein was decreased ( P<0.01). The protein and mRNA levels of ATF3 and CCDC6 were significantly up-regulated (all P<0.05). Conclusions:Ferroptosis may be one of the key mechanisms in the occurrence and development of PMN. In vitro experiments show that ATF3 and CCDC6 are the key genes in the ferroptosis of PMN podocytes, which provides new insights and ideas for the pathogenesis of PMN.

Objective:To explore the characteristics of T cells and natural killer (NK) cells in cerebrospinal fluid of patients with multiple sclerosis (MS).Methods:Cerebrospinal fluids from patients with relapsing-remitting multiple sclerosis (RRMS) and healthy controls attending the Second Hospital of Lanzhou University from January 2023 to October 2024 were collected and analyzed by single-cell RNA sequencing (scRNA-seq) and flow cytometry, and T and NK cell characteristics were summarized and compared in the two groups. The proposed time-series analysis was used to explore the differentiation trajectories of T cells and NK cells.Results:A total of 3 patients with RRMS and 3 healthy controls underwent scRNA-seq, and 6 patients with RRMS and 4 healthy controls underwent flow cytometry. The composition of cerebrospinal fluid T and NK cell subtypes was similar in the RRMS group and the healthy control group, but the proportion of the cell subtypes differed. The RRMS group exhibited significantly higher frequencies of CD4 +Naive T- IL7R [1 529/9 055(16.89%) vs 1 423/9 910(14.36%), χ2=22.980, P<0.001], CD4 +Naive T- CCR7 [1 573/9 055(17.37%) vs 948/9 910(9.57%), χ2=250.114, P<0.001], and CD4 +Naive T- LTB [1 369/9 055(15.12%) vs 1 079/9 910(10.89%), χ2=75.336, P<0.001] subsets compared to controls. Conversely, the control group demonstrated greater proportions of CD4 +Th1- GZMA [1 255/9 910(12.66%) vs 719/9 055(7.94%), χ2=113.213, P<0.001] and CD8 +Tem- GZMK cells [1 607/9 910(16.22%) vs 1 232/9 055(13.61%), χ2=25.326, P<0.001] than the RRMS group. The transcription factors and gene expression of each T cell subtype were also different between the 2 groups, and CD4 initial T cells and CD8 effector T cells were located at the beginning and the end of the differentiation trajectory, respectively. Conclusions:The cerebrospinal fluid of MS patients is characterized by increased expression of genes involved in T cell differentiation and over-activation of immune cells.

Objective:To explore the characteristics of T cells and natural killer (NK) cells in cerebrospinal fluid of patients with multiple sclerosis (MS).Methods:Cerebrospinal fluids from patients with relapsing-remitting multiple sclerosis (RRMS) and healthy controls attending the Second Hospital of Lanzhou University from January 2023 to October 2024 were collected and analyzed by single-cell RNA sequencing (scRNA-seq) and flow cytometry, and T and NK cell characteristics were summarized and compared in the two groups. The proposed time-series analysis was used to explore the differentiation trajectories of T cells and NK cells.Results:A total of 3 patients with RRMS and 3 healthy controls underwent scRNA-seq, and 6 patients with RRMS and 4 healthy controls underwent flow cytometry. The composition of cerebrospinal fluid T and NK cell subtypes was similar in the RRMS group and the healthy control group, but the proportion of the cell subtypes differed. The RRMS group exhibited significantly higher frequencies of CD4 +Naive T- IL7R [1 529/9 055(16.89%) vs 1 423/9 910(14.36%), χ2=22.980, P<0.001], CD4 +Naive T- CCR7 [1 573/9 055(17.37%) vs 948/9 910(9.57%), χ2=250.114, P<0.001], and CD4 +Naive T- LTB [1 369/9 055(15.12%) vs 1 079/9 910(10.89%), χ2=75.336, P<0.001] subsets compared to controls. Conversely, the control group demonstrated greater proportions of CD4 +Th1- GZMA [1 255/9 910(12.66%) vs 719/9 055(7.94%), χ2=113.213, P<0.001] and CD8 +Tem- GZMK cells [1 607/9 910(16.22%) vs 1 232/9 055(13.61%), χ2=25.326, P<0.001] than the RRMS group. The transcription factors and gene expression of each T cell subtype were also different between the 2 groups, and CD4 initial T cells and CD8 effector T cells were located at the beginning and the end of the differentiation trajectory, respectively. Conclusions:The cerebrospinal fluid of MS patients is characterized by increased expression of genes involved in T cell differentiation and over-activation of immune cells.

Objective:To identify and validate the key genes of ferroptosis in phospholipase A2 receptor (PLA2R) associated membranous nephropathy through bioinformatics analysis and in vitro experiments, and to explore the potential role of ferroptosis in PLA2R associated membranous nephropathy (PMN). Methods:The GSE115857 dataset obtained by retrieving the Gene Expression Omnibus (GEO) database and the ferroptosis-related genes obtained by retrieving the FerrDb database were intersected. The intersected genes were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key ferroptosis genes associated with PMN were identified by intersecting genes selected using support vector machines-recursive feature elimination and least absolute shrinkage and selection operator regression. The results were validate by real-time PCR, cell counting kit-8, Western blotting and immunofluorescence in human renal podocyte line AB 8/13 from both the control group and model group.Results:A total of 25 genes related to ferroptosis of PMN were obtained, and GO and KEGG analysis showed that these genes were mainly involved in cell ferroptosis metabolism. The key ferroptosis genes of PMN obtained by machine learning method were activating transcription factor 3 ( ATF3) and coiled coil domain containing 6 ( CCDC6). The results of in vitro experiments showed that the human renal podocyte line AB 8/13 in the model group was significantly deformed and retracted compared with the control group. The surface area density of foot processes was significantly reduced, and the podocyte cytoskeleton was allosteric. The morphology of F-actin was disordered and the expression of synaptopodin was decreased. The cell proliferation activity was significantly decreased ( P<0.05). The expression of PLA2R protein was increased ( P<0.05), and the expression of GPX4 protein was decreased ( P<0.01). The protein and mRNA levels of ATF3 and CCDC6 were significantly up-regulated (all P<0.05). Conclusions:Ferroptosis may be one of the key mechanisms in the occurrence and development of PMN. In vitro experiments show that ATF3 and CCDC6 are the key genes in the ferroptosis of PMN podocytes, which provides new insights and ideas for the pathogenesis of PMN.

As the incidence of chronic kidney disease (CKD) increases year by year, the number of patients with CKD is increasing year by year before or during pregnancy. Pregnancy management is increasingly important for patients with kidney disease, and women with CKD face a number of challenges during pregnancy, such as the risk of kidney disease progression, fetal growth restriction, the potential teratogenicity of medications, and an increased risk of complications such as preeclampsia and preterm birth. In order to ensure a good outcome for pregnant women and the fetus, it is necessary for multidisciplinary physicians in the department of nephrology, obstetrics, reproductive health and other departments to assess the risk of pregnancy in patients with CKD in a standardized manner, adjust medication in a timely manner, and closely monitor to detect maternal and fetal complications early, stabilize the patient′s condition, and determine the safest time for pregnancy and delivery.

"Early clinical practice, multiple clinical practice and repeated clinical practice" is the mainstream of the reform of higher medical education in the 21st century. Based on the shortages of the current medical education model, a university started to reform from both theoretical teaching and practical teaching, and constructed a teaching model of "early clinical practice, multiple clinical practice and repeated clinical practice" that fits China's reality. And the teaching effect was evaluated questionnaire survey and OSCE tests. The results showed that this teaching model could promote the all-round development of medical students' knowledge, ability and quality.

Objective To investigate the levels of trace elements in preschool children in Guangzhou and their relationships with nutritional status, and to provide a scientific basis for guiding children's nutritional diet and timely and reasonable supplementation of trace elements. Methods A cross-sectional study was used to perform physical examinations and collect blood samples from 5 002 children who were examined in the Department of Pediatric Health of a Maternal and Child Health Hospital in Guangzhou in 2018. The levels of trace elements including iron, zinc and copper were detected by atomic absorption spectrometer. Results The median (upper and lower quartile) levels of iron, zinc and copper for preschool children in a district of Guangzhou were 7.80 (7.39, 8.19) mmol / L, 69.0 (61.0, 76.5) μmol / L, 18.6 (15.9, 21.4) μmol / L. There was no significant difference in the levels of iron, zinc and copper between different genders (P>0.05). The levels of iron and zinc increased significantly with the age of children (P=0.000). There was no significant difference in the level of copper in different ages (P>0.05). The prevalence rates of iron, zinc and copper deficiency in preschool children were 14.93%, 21.93% and 0.24%, respectively, and there was no significant difference between different genders (P>0.05).There were no significant differences in the prevalence of iron and copper deficiency in different ages (P>0.05), while the zinc deficiency rate decreased with the age of children (P = 0.000). (P<0.05). The levels of iron and zinc in overweight children, and those suffering from malnutrition,and obesity were lower than those in normal children. There was no statistically significant difference in the levels of copper in children with different nutritional status (P>0.05). The prevalence rates of iron and zinc deficiency in children with malnutrition, overweight and obesity were higher than those in normal children. There was no significant difference in the prevalence of copper deficiency in children with different nutritional status (P>0.05). Conclusion The prevalence of iron, zinc deficiency is high in preschool children in Guangzhou. Preschool children with malnutrition, overweight and obesity are more prone to iron and zinc deficiency.

Objective To investigate the influencing factors of quality in contrast-enhanced ultrasound (CEUS) perfusion analysis for hepatocellular carcinoma (HCC) and the reproducibility of using CEUS perfusion analysis on HCC.Methods A total of 136 HCC lesions were undergone CEUS perfusion analysis.Maximum intensity (Imax),rise time (RT),time to peak (TTP),mean transit time (mTF) and quality of fit (QOF) of HCC lesion,top-enhanced region inside HCC lesion and adjacent liver parenchyma were measured.The quality of perfusion analysis was classified into three grades " Good" (QOF ＞ 75％),"Common" [QOF (50％-75％)],and " Poor" (QOF ＜ 75％).Ultrasound machine,patients' age,tumor depth,tumor size and tumor location were recorded and compared among the three groups.20 consecutive patients with 20 HCC lesions were received CEUS perfusion analysis by two operators and another 20 consecutive HCC lesions were received CEUS perfusion analysis by one operator at different time.Intra-class correlation coefficient (ICC) was used to evaluate the reproducibility of inter-and intra-observer.Results Fifty-six HCC lesions were classified as " Good",39 as " Common" and 39 as " Poor",respectively.Tumor size (P =0.015) and tumor location (P =0.041) were significantly different among the three groups.Tumor size ＞ 3.0 cm and tumor located in S3,S4b,S5,S6 were apt to gain a better QOF.No significant difference was found for the different ultrasound machine,tumor depth and patients' age.For all CEUS perfusion analysis parameters of both lesion and adjacent parenchyma,intra-class correlation coefficient of inter-and intra-observer were higher than 0.90,the reproducibility of CEUS perfusion analysis was good.For Imax,RT and TTP in top-enhanced region inside HCC lesion,ICC of inter-and intra-observer was all higher than 0.90 (good).For mITT in highest enhanced area inside HCC lesion,ICC of inter-and intra-observer were 0.459 (poor),0.609 (common),respectively.Conclusions The parameters of HCC quantitative perfusion analysis were reproducible in peripheral liver parenchyma and tumor,but the mTT repeatability was poor in the highest enhancement area of the tumor.The size and location of the lesion were the factors affecting quantitative analysis.The quantitative analysis of ＞ 3.0 cm and the tumor located in S3,S4b,S5,S6 in the liver were satisfactory.

Objective:To evaluate a Meta-analysis of randomized controlled trials ( RCTs) in multiple sclerosis ( MS) patients to evaluate the efficacy of vitamin D as add-on therapy. Methods: Searched Pubmed,EMbase,the Cochrane Library,CNKI,Wanfang Data base and so on up to february 2016 using the keywords:multiple sclerosis or MS and the drug names:vitamin D orCholecalciferol. Two authors independently selected the articles and extracted the data. We performed meta-analysis using Review Manager ( RevMan) version 5. 3 software. Results:Four RCTs with a total of 247 patients were selected.①Compared to the placebo, the EDSS score[MD=-0. 33,95% Confidence interval (CI)= (0. 68,0. 01),P=0. 05],the annual relapse rate[MD=-0. 08, 95%CI=(-0.37,0.21),P=0.60]and the number of gadolinium-enhancing lesions[MD=-0.16,95%CI=(-0.57,0.25),P=0. 45] showed no significant difference at 12 months,meanwhile the EDSS score[MD=-0. 48,95%CI=(0. 87,-0. 09),P=0. 02] and the annual relapse rate[MD=-0. 27,95%CI=(-0. 52,-0. 02),P=0. 03] were significantly less in the vitamin D group at 24 months.②Safety evaluation:There was no hypercalcaemia in vitamin D treated patients in each studies,main adverse events reported were diarrhoea, fever, constipation, dyspepsia, headache and so on. These symptoms were mild, after stopping drug can relieve the general. Conclusion: Vitamin D as an added in the treatment of MS showed as same as the placebo in some clinical indicators. However,after a longer treatment, the clinical indicators were significantly lower in the vitamin D group. Due to limited quantity and quality of the included studies,further larger and more prolonged studies are merited to verify the above conclusion.

Objective To compare the features of different heating modes cauterization mode and standard ablation mode of Cool-tip on coagulation zone and thermal field distribution in order to provide references for focused and accurate vascular occlusion and explore the primary efficacy of Cool-tip applied to intrahepatic vascular occlusion in living dogs Methods Ex-vivo porcine livers were ablated for 2 4 6 8 minites with cauterization mode and standard ablation mode respectively and the long- and short-axis diameters in the greatest dimension were compared The tissue temperature around the tip midpoint and end of the exposed part of radiofrequency electrode were measured under the circumstance of 4 min ablation by the two modes respectively Animal experiment the intrahepatic vessels of 2 Beagles were ablated with cauterization mode one procedure for each animal and contrast-enhanced ultrasound was used to evaluate the perfusion change of liver tissue Gross anatomy and pathological examination of the liver was performed after 1 week Results In the 2 min 4 min 6 min and 8 min ablation the coagulation volume of cauterization mode and standard ablation mode were 1 01 ± 0 41 cm 3 vs 2 95 ± 0 74 cm 3 1 47 ± 0 33 cm 3 vs 5 03±1 06 cm 3 2 29±0 49 cm 3 vs 9 23±2 53 cm 3 2 70±0 24 cm 3 vs 1 5 89±0 77 cm 3 The coagulation volume was significant smaller in cauterization mode than in standard ablation mode P <0 05 for all The peak temperature of cauterization mode presented on the tip of electrode which averaged 94 4℃ in maximum and maintain around 70 ℃ in the equilibrium phase The peak temperature of standard mode presented on the midpoint of the electrode which periodically variated between 70 ℃- 100 ℃ The result of the animal experiment showed that cauterization mode could be used to occlude the intrahepatic vessel in vivo which induced the ischemia and necrosis in the corresponding area Conclusions Cauterization mode and standard alation mode of Cool-tip have different characteristic in terms of coagulation zone and thermal distribution and carterization mode may have a promising application in intrahepatic vascular occlusion.