ObjectiveTo observe the clinical effect of Xiaozhi Hugan capsules in treating patients with non-alcoholic steatohepatitis （NASH） combined with phlegm-dampness and blood stasis syndrome and its effects on serum interleukin-6 （IL-6） and monocyte chemoattractant protein-1 （MCP-1）. MethodsA total of 124 patients with NASH combined with phlegm-dampness and blood stasis syndrome who were admitted to the Department of Spleen， Stomach， and Hepatobiliary Diseases， the First Affiliated Hospital of Henan University of Chinese Medicine from July 2020 to December 2022 were selected. According to the random number table method， patients were randomly divided into an observation group （62 cases） and a control group （62 cases）. The treatment duration was 6 months. The observation group received Xiaozhi Hugan capsules orally， while the control group received polyene phosphatidylcholine capsules. The efficacy indicators included alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， controlled attenuation parameter （CAP）， liver stiffness measurement （LSM）， traditional Chinese medicine （TCM） syndrome scores （discomfort/dull pain/distending pain in liver region， fatigue， etc.）， body mass index （BMI）， waist-to-height ratio （WHtR）， high-density lipoprotein cholesterol （HDL-C）， total cholesterol （TC）， triglycerides （TG）， homeostatic model assessment for insulin resistance （HOMA-IR） ［including fasting blood glucose （FBG） and fasting insulin level （INS）］， free fatty acids （FFA）， IL-6， and MCP-1. Adverse drug reactions were recorded. ResultsAfter treatment， the total effective rate in the observation group was 92.3% （48/52）， while that in the control group was 75.5% （39/49）. The total effective rate in the observation group was higher than that in the control group （χ²=5.339， P<0.05）. After treatment， the TCM syndrome scores in both groups were significantly reduced （P<0.05）， and the post-treatment scores in the observation group were better than those in the control group （P<0.05）. After treatment， the levels of ALT， AST， TC， FFA， fasting insulin （FINS）， HOMA-IR， MCP-1， IL-6， CAP， LSM， BMI， and WHtR were decreased （P<0.05） significantly in both groups， and the observation group showed superior improvement in the above indicators compared to the control group （P<0.05）. The observation group exhibited significant reductions in TG and FBG （P<0.05） and an increase in HDL-C （P<0.05）， while no significant changes were observed in the control group. The observation group was superior to the control group after treatment （P<0.05）. No severe adverse reactions occurred in either group during the treatment. ConclusionXiaozhi Hugan capsules have significant clinical efficacy in treating patients with NASH combined with phlegm-dampness and blood stasis syndrome. It reduces hepatic steatosis， lowers liver stiffness， inhibits the expression of serum inflammatory factors， and alleviates liver inflammation. No obvious adverse reactions occur， suggesting it is suitable for clinical application.

ObjectiveTo observe the clinical effect of Xiaozhi Hugan capsules in treating patients with non-alcoholic steatohepatitis （NASH） combined with phlegm-dampness and blood stasis syndrome and its effects on serum interleukin-6 （IL-6） and monocyte chemoattractant protein-1 （MCP-1）. MethodsA total of 124 patients with NASH combined with phlegm-dampness and blood stasis syndrome who were admitted to the Department of Spleen， Stomach， and Hepatobiliary Diseases， the First Affiliated Hospital of Henan University of Chinese Medicine from July 2020 to December 2022 were selected. According to the random number table method， patients were randomly divided into an observation group （62 cases） and a control group （62 cases）. The treatment duration was 6 months. The observation group received Xiaozhi Hugan capsules orally， while the control group received polyene phosphatidylcholine capsules. The efficacy indicators included alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， controlled attenuation parameter （CAP）， liver stiffness measurement （LSM）， traditional Chinese medicine （TCM） syndrome scores （discomfort/dull pain/distending pain in liver region， fatigue， etc.）， body mass index （BMI）， waist-to-height ratio （WHtR）， high-density lipoprotein cholesterol （HDL-C）， total cholesterol （TC）， triglycerides （TG）， homeostatic model assessment for insulin resistance （HOMA-IR） ［including fasting blood glucose （FBG） and fasting insulin level （INS）］， free fatty acids （FFA）， IL-6， and MCP-1. Adverse drug reactions were recorded. ResultsAfter treatment， the total effective rate in the observation group was 92.3% （48/52）， while that in the control group was 75.5% （39/49）. The total effective rate in the observation group was higher than that in the control group （χ²=5.339， P<0.05）. After treatment， the TCM syndrome scores in both groups were significantly reduced （P<0.05）， and the post-treatment scores in the observation group were better than those in the control group （P<0.05）. After treatment， the levels of ALT， AST， TC， FFA， fasting insulin （FINS）， HOMA-IR， MCP-1， IL-6， CAP， LSM， BMI， and WHtR were decreased （P<0.05） significantly in both groups， and the observation group showed superior improvement in the above indicators compared to the control group （P<0.05）. The observation group exhibited significant reductions in TG and FBG （P<0.05） and an increase in HDL-C （P<0.05）， while no significant changes were observed in the control group. The observation group was superior to the control group after treatment （P<0.05）. No severe adverse reactions occurred in either group during the treatment. ConclusionXiaozhi Hugan capsules have significant clinical efficacy in treating patients with NASH combined with phlegm-dampness and blood stasis syndrome. It reduces hepatic steatosis， lowers liver stiffness， inhibits the expression of serum inflammatory factors， and alleviates liver inflammation. No obvious adverse reactions occur， suggesting it is suitable for clinical application.

Objective To explore the potential mechanism of Xinnao Maikang in improving ApoE-/-atherosclerosis mice based on PPARγ/LXRα/ABCA1 pathway.Methods Totally 50 ApoE-/-mice were randomly divided into model group,atorvastatin group,Xinnao Maikang high-,medium-and low-dosage groups,and fed with high-fat diet for 12 weeks to establish atherosclerosis model.Another 10 C57BL/6J mice were selected as normal group,and atorvastatin group was given atorvastatin calcium suspension by intragastric administration,Xinnao Maikang high-,medium-and low-dosage groups were given Xinnao Maikang Decoction 56,28,14 g/kg by intragastric administration,the normal group and model group were given equal volume of normal saline by intragastric administration for 8 weeks.Liver index of mice was detected,serum lipid level of mice was detected by automatic biochemical analyzer,TC and TG contents in liver tissue were detected by biochemical kit,morphology of aortic intima and liver tissue were observed by HE staining,lipid deposition in liver tissue was observed by oil red O staining,the expressions of PPARγ,LXRα,ABCA1,ABCG1 and SR-BⅠ in liver tissue were detected by Western blot.Results Compared with the normal group,the liver index in the model group significantly increased(P<0.05),the contents of serum TC,TG and LDL-C significantly increased(P<0.05),and the contents of HDL-C significantly decreased(P<0.05),the contents of TC and TG in liver tissue increased(P<0.05),the aortic wall was thickened,the number of subcutaneous foam cells increased,the arrangement of liver cells were disaffected,and a large number of fat vacuoles could be seen,the protein expressions of PPARγ,LXRα,ABCA1,ABCG1 and SR-BⅠ in liver tissue significantly decreased(P<0.01).Compared with the model group,the liver index of Xinnao Maikang high-,medium-and low-dosage groups and atorvastatin group significantly decreased(P<0.05),the serum contents of TC,TG and LDL-C significantly decreased(P<0.05),and the HDL-C contents significantly increased(P<0.05),the contents of TC and TG in liver tissue significantly decreased(P<0.05),the aortic wall thickened and the subcutaneous foam cells decreased,the liver cells were arranged neatly,and fat degeneration was reduced to varying degrees,the protein expressions of PPARγ,LXRα,ABCA1,ABCG1 and SR-BⅠ in liver tissue of Xinnao Maikang high-dosage group and atorvastatin group significantly increased(P<0.05,P<0.01).Conclusion Xinnao Maikang can decrease blood lipid level,reduce atherosclerotic plaque and reduce lipid deposition in liver of atherosclerotic mice,and its mechanism may be related to regulating PPARγ/LXRα/ABCA1 pathway,regulating lipid metabolism,promoting reverse cholesterol transport.

Objective:To explore the clinical characteristics and gene variant of a fetus with Farber lipogranulomatosis caused by ASAH1 gene variant. Methods:A fetus with Farber lipogranulomatosis caused by ASAH1 gene variant diagnosed at Women and Children′s Hospital of Ningbo University in August 2024 was selected as the subject. Clinical data and abortion tissue samples of the fetus and peripheral blood samples of its parents were collected for whole exome sequencing (WES). Sanger sequencing validation and bioinformatics analysis were performed on candidate variants. This study was approved by Women and Children′s Hospital of Ningbo University (Ethics No. EC2020-048). Results:Generalized skin oedema, pericardial effusion, right pleural effusion and increased bowel echogenicity of the fetus were founded by prenatal ultrasound. WES revealed that the fetus has harbored a homozygous c. 101C>A(p.Ser34Ter) variation in exon 2 of the ASAH1 gene. Sanger sequencing confirmed that both parents carry the heterozygous nonsense variation c. 101C>A (p.Ser34Ter) in ASAH1 gene, which has not been included in databases such as HGMD, ClinVar, 1000 Genomes, ExAC, dbSNP, and gnomAD. Based on the Standards and Guidelines for the Interpretation of Sequence Variants of the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic (PM2_Supporting+ PVS1+ PM3_Supporting). The AlphaFold3 model protein structure prediction reveals that the c. 101C>A variant caused the premature appearance of a termination codon, resulting in only a small partial α-helix structure in the N-terminal of the encoded ASAH1 protein, with the complete loss of the α-helix structure in the core domain, which might lead to the loss of function of this protein. Conclusion:The c. 101C>A(p.Ser34Ter) variant of the ASAH1 gene probably underlay the Farber lipogranulomatosis with hydrops fetalis in this fetus. The newly discovered c. 101C>A(p.Ser34Ter) variant has enriched the mutational spectrum of Farber lipogranulomatosis.

Objective:To explore the genetic etiology of a child with brain small vessel disease 1 with ocular anomalies.Methods:A child who was adwstted to Ningbo Women and Children′s Hospital on May 28, 2022, was selected for the study. Clinical data were collected, and peripheral blood samples from the child and her parents were obtained for genomic DNA extraction. Whole exome sequencing (WES) was performed to screen for pathogenic variants. Candidate variants were validated via Sanger sequencing and subjected to bioinformatic analysis. This study was approved by the Medical Ethics Committee of Ningbo Women and Children′s Hospital (Ethics No. EC2020-048).Results:① The child was a 7-year-old female with a diagnosis of epilepsy. ② WES revealed that she has carried a heterozygous missense variant in the COL4A1 gene: c. 1792G>A (p.Gly598Ser). Sanger sequencing confirmed that her parents both had the wild-type genotype for this variant. Based on American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants, the variant were predicted to be a likely pathogenic (PS2+ PM1+ PM2_Supporting+ PP3). ③ Bioinformatics predicted that amino acid 598 was highly conserved in different species, formed hydrogen bond with Asp599 after becoming Ser598. Conclusion:The heterozygous missense variant of the COL4A1 gene c. 1792G>A (p.G598S) could be the pathogenic cause of this child with Brain small vessel disease 1 with ocular anomalies.

Objective:To explore the genetic characteristics of a fetus affected with Structural heart defects and renal anomalies syndrome (SHDRA).Methods:A pedigree with SHDRA (fetus and the parents) who had visited the Affiliated Women and Children′s Hospital of Ningbo University in April 2023 was selected as the study subject. Clinical data of the family were collected. A total of 10 mL of amniotic fluid cells from the fetus and 5 mL of peripheral blood samples from the parents were collected for genomic DNA extraction. Trio whole-exome sequencing (Trio-WES) was performed, and Sanger sequencing was used to validate candidate variants in the family. The identified variants were classified according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereinafter referred to as the " ACMG Guidelines). Relevant research literature on SHDRA in domestic and international databases were searched for literature review. This study was approved by the Affiliated Women and Children′s Hospital of Ningbo University (Ethics No. EC2023-094).Results:①In this family, prenatal ultrasound at 18 weeks of gestation revealed left renal multicystic dysplasia in the fetus. After birth, the infant exhibited an ostium secundum atrial septal defect, patent ductus arteriosus, and left renal multicystic dysplasia. Trio-WES revealed that the fetus had carried c. 344dup(p.L116Afs*32) and c. 90_104dup(p.Ala31_Ala35dup) compound heterozygous variants in the TMEM260 gene, which were respectively inherited from its father and mother. According to the ACMG guidelines, the c. 344dup(p.L116Afs*32) and c. 90_104dup (p.Ala31_Ala35dup) variants were classified as pathogenic (PM2_Supporting+ PVS1+ PP4) and likely pathogenic (PM2_Supporting+ PM4+ PM3+ PP4), respectively. ②According to the literature search strategy set for this study, a total of 6 literature was retrieved, involving 25 SHDRA patients from 20 families. Together with the patients in this study, there were 14 TMEM260 gene variants, most of which were frameshift variants (7 types) and had located in exons 3, 11 and 13. The main clinical features of SHDRA were congenital heart malformation, renal abnormality and neurodevelopmental abnormality, and there was a lack of genotype-phenotype correlation. Conclusion:The c. 344dup(p.L116Afs*32) and c. 90_104dup(p.Ala31_Ala35dup) variants of the TMEM260 gene probably underlay the SHDRA in this family. Above finding has provided a basis for clinical diagnosis and genetic counseling for the family.

Lung transplantation is the ultimate therapeutic option for end-stage pulmonary diseases such as interstitial pneumonia, chronic obstructive pulmonary disease and pneumoconiosis. Currently, the shortage of allogeneic lung donors significantly limits the opportunity for end-stage lung disease patients to receive lung transplantation. In recent years, with the rapid development of biomedical engineering technologies, especially the major breakthroughs in genetic modification and cloning, xenogeneic lung transplantation has shown important potential for clinical translation. Among them, genetically modified pigs have become the most promising xenogeneic lung source due to the close similarity of organ size and physiological characteristics to humans, and the ability to perform targeted gene knockouts (such as α-Gal antigen knockout) to reduce the occurrence of hyperacute rejection. This article focuses on the research progress of porcine xenogeneic lung transplantation, systematically reviews the latest achievements and challenges in animal experiments and human trials, and introduces the technical experience accumulated by Shenzhen Third People's Hospital in the porcine-to-monkey xenogeneic lung transplantation model, in the hope of providing practical references for future research in this field.

OBJECTIVE: To explore the clinical characteristics and gene variant of a fetus with Farber lipogranulomatosis caused by ASAH1 gene variant. METHODS: A fetus with Farber lipogranulomatosis caused by ASAH1 gene variant diagnosed at Women and Children's Hospital of Ningbo University in August 2024 was selected as the subject. Clinical data and abortion tissue samples of the fetus and peripheral blood samples of its parents were collected for whole exome sequencing (WES). Sanger sequencing validation and bioinformatics analysis were performed on candidate variants. This study was approved by Women and Children's Hospital of Ningbo University (Ethics No. EC2020-048). RESULTS: Generalized skin oedema, pericardial effusion, right pleural effusion and increased bowel echogenicity of the fetus were founded by prenatal ultrasound. WES revealed that the fetus has harbored a homozygous c.101C>A (p.Ser34Ter) variation in exon 2 of the ASAH1 gene. Sanger sequencing confirmed that both parents carry the heterozygous nonsense variation c.101C>A (p.Ser34Ter) in ASAH1 gene, which has not been included in databases such as HGMD, ClinVar, 1000 Genomes, ExAC, dbSNP, and gnomAD. Based on the Standards and Guidelines for the Interpretation of Sequence Variants of the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic (PM2_Supporting+PVS1+PM3_Supporting). The AlphaFold3 model protein structure prediction reveals that the c.101C>A variant caused the premature appearance of a termination codon, resulting in only a small partial α-helix structure in the N-terminal of the encoded ASAH1 protein, with the complete loss of the α-helix structure in the core domain, which might lead to the loss of function of this protein. CONCLUSION The c.101C>A (p.Ser34Ter) variant of the ASAH1 gene probably underlay the Farber lipogranulomatosis with hydrops fetalis in this fetus. The newly discovered c.101C>A (p.Ser34Ter) variant has enriched the mutational spectrum of Farber lipogranulomatosis.
Humans ; Female ; Pregnancy ; Acid Ceramidase/chemistry* ; Farber Lipogranulomatosis/diagnostic imaging* ; Fetus ; Exome Sequencing ; Adult

OBJECTIVE: To explore the genetic characteristics of a fetus affected with Structural heart defects and renal anomalies syndrome (SHDRA). METHODS: A pedigree with SHDRA (fetus and the parents) who had visited the Affiliated Women and Children's Hospital of Ningbo University in April 2023 was selected as the study subject. Clinical data of the family were collected. A total of 10 mL of amniotic fluid cells from the fetus and 5 mL of peripheral blood samples from the parents were collected for genomic DNA extraction. Trio whole-exome sequencing (Trio-WES) was performed, and Sanger sequencing was used to validate candidate variants in the family. The identified variants were classified according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereinafter referred to as the "ACMG Guidelines). Relevant research literature on SHDRA in domestic and international databases were searched for literature review. This study was approved by the Affiliated Women and Children's Hospital of Ningbo University (Ethics No. EC2023-094). RESULTS: In this family, prenatal ultrasound at 18 weeks of gestation revealed left renal multicystic dysplasia in the fetus. After birth, the infant exhibited an ostium secundum atrial septal defect, patent ductus arteriosus, and left renal multicystic dysplasia. Trio-WES revealed that the fetus had carried c.344dup (p.L116Afs*32) and c.90_104dup (p.Ala31_Ala35dup) compound heterozygous variants in the TMEM260 gene, which were respectively inherited from its father and mother. According to the ACMG guidelines, the c.344dup (p.L116Afs*32) and c.90_104dup (p.Ala31_Ala35dup) variants were classified as pathogenic (PM2_Supporting+PVS1+PP4) and likely pathogenic (PM2_Supporting+PM4+PM3+PP4), respectively. According to the literature search strategy set for this study, a total of 6 literature was retrieved, involving 25 SHDRA patients from 20 families. Together with the patients in this study, there were 14 TMEM260 gene variants, most of which were frameshift variants (7 types) and had located in exons 3, 11 and 13. The main clinical features of SHDRA were congenital heart malformation, renal abnormality and neurodevelopmental abnormality, and there was a lack of genotype-phenotype correlation. CONCLUSION The c.344dup (p.L116Afs*32) and c.90_104dup (p.Ala31_Ala35dup) variants of the TMEM260 gene probably underlay the SHDRA in this family. Above finding has provided a basis for clinical diagnosis and genetic counseling for the family.
Humans ; Female ; Pedigree ; Membrane Proteins/genetics* ; Male ; Heart Defects, Congenital/genetics* ; Kidney/abnormalities* ; Pregnancy ; Adult ; Kidney Diseases/congenital* ; Exome Sequencing ; Mutation ; Genetic Testing

OBJECTIVE: To explore the genetic etiology of a child with Brain small vessel disease 1 with ocular anomalies. METHODS: A child who was admitted to Ningbo Women and Children's Hospital on May 28, 2022 was selected for the study. Clinical data were collected, and peripheral blood samples from the child and her parents were obtained for genomic DNA extraction. Whole exome sequencing (WES) was performed to screen for pathogenic variants. Candidate variants were validated via Sanger sequencing and subjected to bioinformatic analysis. This study was approved by the Medical Ethics Committee of Ningbo Women and Children's Hospital (Ethics No. EC2020-014). RESULTS: The child was a 7-year-old female with a diagnosis of epilepsy. WES revealed that she has carried a heterozygous missense variant in the COL4A1 gene: c.1792G>A (p.Gly598Ser). Sanger sequencing confirmed that her parents both had the wild-type genotype for this variant. Based on American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants, the variant were predicted to be a likely pathogenic (PS2+PM1+PM2_Supporting+PP3). Bioinformatics predicted that amino acid 598 was highly conserved in different species, formed hydrogen bond with Asp599 after becoming Ser598. CONCLUSION The heterozygous missense variant of the COL4A1 gene c.1792T>C (p.G598S) could be the pathogenic cause of this child with Brain small vessel disease 1 with ocular anomalies.
Humans ; Female ; Child ; Collagen Type IV/genetics* ; Eye Abnormalities/genetics* ; Exome Sequencing ; Mutation, Missense ; Cerebral Small Vessel Diseases/genetics*