Objective Through molecular genetics analysis of a calculi family lineage,this study aims to explore its pathogenesis and the association between genotypes and phenotypes.Methods A retrospective analysis was conducted on a calculi family lineage admitted to Tongji Hospital,affiliated with Tongji Medical College,Huazhong University of Science and Technology.Clinical data and peripheral blood samples of the affected children and some family members were collected.Whole exome sequencing was performed,followed by Sanger sequencing to validate the candidate variants.Results Among the 38 family members across four generations,10 members were diagnosed with calculi disease.The second generation,member 2(Ⅱ-2),third generation,member 2(Ⅲ-2),and third generation,member 4(Ⅲ-4)suffered from recurrent multiple kidney stones and gallstones.The second generation,member 6(Ⅱ-6),second generation,member 13(Ⅱ-13),and third generation,member 5(Ⅲ-5)had recurrent multiple kidney stones alone,while first generation,member 2(Ⅰ-2),second generation,member 4(Ⅱ-4),second generation,member 8(Ⅱ-8),and second generation,member 11(Ⅱ-11)only had gallstones.No other family members exhibited any signs of kidney or gallbladder involvement.Ⅱ-2 was diagnosed in 2018 with end-stage renal disease stage 5,grade 3 hypertension and gallstones,urinary amino acid high-performance liquid chromatography analysis indicated elevated urinary cystine.This member had a history of recurrent multiple kidney stones and recurrent urinary tract infections for over 30 years,with multiple histories of ureteroscopic stone removal.Genetic analysis revealed that Ⅱ-2,Ⅲ-2,Ⅲ-4,Ⅲ-5 and Ⅳ-1 all carry a heterozygous mutation in exon 10 of the solute carrier family 3 member 1(SLC3A1)gene,c.1889G＞A(p.Gly630Asp).The third generation,member 1(Ⅲ-1),and fourth generation,member 2(Ⅳ-2),are wild type.This mutation shows a phenomenon of family co-segregation.Conclusions The heterozygous mutation of SLC3A1 gene,c.1889G＞A,may be the genetic cause of calculi disease in multiple members of this family lineage.Recurrent multiple kidney stones and/or gallstones require high attention to genetic etiology.It is recommended to perform genetic analysis on calculi family lineages and patients with early-onset calculi disease.

This study investigated the role of the nuclear factor of activated T cells c3 (NFATc3) in vascular smooth muscle cells (VSMCs) during aortic aneurysm and dissection (AAD) progression and the underlying molecular mechanisms. Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD. VSMC-NFATc3 deletion reduced thoracic AAD (TAAD) and abdominal aortic aneurysm (AAA) progression in mice, contrary to VSMC-NFATc3 overexpression. VSMC-NFATc3 deletion reduced extracellular matrix (ECM) degradation and maintained the VSMC contractile phenotype. Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9 (MMP9) and MMP2, promoting ECM degradation and AAD development. NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2 (eEF2) and inhibiting its phosphorylation in the VSMC cytoplasm. Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice. Cabamiquine-targets eEF2 and inhibits protein synthesis-inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice. VSMC-NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development, making it a novel potential therapeutic target for preventing and treating AAD.

Objective Through molecular genetics analysis of a calculi family lineage,this study aims to explore its pathogenesis and the association between genotypes and phenotypes.Methods A retrospective analysis was conducted on a calculi family lineage admitted to Tongji Hospital,affiliated with Tongji Medical College,Huazhong University of Science and Technology.Clinical data and peripheral blood samples of the affected children and some family members were collected.Whole exome sequencing was performed,followed by Sanger sequencing to validate the candidate variants.Results Among the 38 family members across four generations,10 members were diagnosed with calculi disease.The second generation,member 2(Ⅱ-2),third generation,member 2(Ⅲ-2),and third generation,member 4(Ⅲ-4)suffered from recurrent multiple kidney stones and gallstones.The second generation,member 6(Ⅱ-6),second generation,member 13(Ⅱ-13),and third generation,member 5(Ⅲ-5)had recurrent multiple kidney stones alone,while first generation,member 2(Ⅰ-2),second generation,member 4(Ⅱ-4),second generation,member 8(Ⅱ-8),and second generation,member 11(Ⅱ-11)only had gallstones.No other family members exhibited any signs of kidney or gallbladder involvement.Ⅱ-2 was diagnosed in 2018 with end-stage renal disease stage 5,grade 3 hypertension and gallstones,urinary amino acid high-performance liquid chromatography analysis indicated elevated urinary cystine.This member had a history of recurrent multiple kidney stones and recurrent urinary tract infections for over 30 years,with multiple histories of ureteroscopic stone removal.Genetic analysis revealed that Ⅱ-2,Ⅲ-2,Ⅲ-4,Ⅲ-5 and Ⅳ-1 all carry a heterozygous mutation in exon 10 of the solute carrier family 3 member 1(SLC3A1)gene,c.1889G＞A(p.Gly630Asp).The third generation,member 1(Ⅲ-1),and fourth generation,member 2(Ⅳ-2),are wild type.This mutation shows a phenomenon of family co-segregation.Conclusions The heterozygous mutation of SLC3A1 gene,c.1889G＞A,may be the genetic cause of calculi disease in multiple members of this family lineage.Recurrent multiple kidney stones and/or gallstones require high attention to genetic etiology.It is recommended to perform genetic analysis on calculi family lineages and patients with early-onset calculi disease.

[Objective]To summarize Professor ZHANG Yonghua's academic experience in the treatment of panic disorder by using the theory of emotion-pattern differentiation.[Methods]Through follow-up study,reading ancient books and sorting out clinical medical cases,this paper analyzed Professor ZHANG Yonghua's clinical experience in exploring the etiology and pathogenesis of panic disorder from the perspective of emotion,as well as the clinical experience of using emotion-state syndrome differentiation theory in the treatment of panic disorder,accompanied with a medical case.[Results]The etiology of panic disorder is mainly related to the invasion of external evils,emotional stimulation and endowment differences,among which Professor ZHANG Yonghua believes that endowment personality is the basis of this disease.The pathogenesis is mainly related to fiery heat,phlegm obstruction,Qi depression,and lack of heart and Yang,but the first three are mainly mixed with each other to cause disease.According to the clinical characteristics and personality manifestations,panic disorder is divided into four categories:impatience and irritability,worry,sadness and depression,and simple panic.In terms of treatment,the combination of disease differentiation,syndrome differentiation and sentiment differentiation pays attention to flirting,and the treatment is mainly to calm the nerves,as well as clear heat,dissolve phlegm,regulate Qi and warm Yang.The proved case belonged to the type of impatience and irritability-more thinking and more consideration through emotional syndrome differentiation.The treatment was to clear away heat and phlegm,calm the nerves,administered with Chaihu plus Longgu Muli Decoction combined with Anshen Dingzhi Pill,and finally achieved good results.[Conclusion]Professor ZHANG Yonghua's treatment of panic disorder based on the theory of"emotion-state syndrome differentiation"has a significant curative effect,which is worthy of clinical promotion and learning.

[Objective]To summarize Professor ZHANG Yonghua's academic experience in the treatment of panic disorder by using the theory of emotion-pattern differentiation.[Methods]Through follow-up study,reading ancient books and sorting out clinical medical cases,this paper analyzed Professor ZHANG Yonghua's clinical experience in exploring the etiology and pathogenesis of panic disorder from the perspective of emotion,as well as the clinical experience of using emotion-state syndrome differentiation theory in the treatment of panic disorder,accompanied with a medical case.[Results]The etiology of panic disorder is mainly related to the invasion of external evils,emotional stimulation and endowment differences,among which Professor ZHANG Yonghua believes that endowment personality is the basis of this disease.The pathogenesis is mainly related to fiery heat,phlegm obstruction,Qi depression,and lack of heart and Yang,but the first three are mainly mixed with each other to cause disease.According to the clinical characteristics and personality manifestations,panic disorder is divided into four categories:impatience and irritability,worry,sadness and depression,and simple panic.In terms of treatment,the combination of disease differentiation,syndrome differentiation and sentiment differentiation pays attention to flirting,and the treatment is mainly to calm the nerves,as well as clear heat,dissolve phlegm,regulate Qi and warm Yang.The proved case belonged to the type of impatience and irritability-more thinking and more consideration through emotional syndrome differentiation.The treatment was to clear away heat and phlegm,calm the nerves,administered with Chaihu plus Longgu Muli Decoction combined with Anshen Dingzhi Pill,and finally achieved good results.[Conclusion]Professor ZHANG Yonghua's treatment of panic disorder based on the theory of"emotion-state syndrome differentiation"has a significant curative effect,which is worthy of clinical promotion and learning.

Objective:To analyze and summarize the clinical, genotypic and pathological characteristics of children with PAX2 gene mutation in China, and to provide information for the monitoring, treatment and prognosis of the disease. Methods:It was a case series analysis study. The clinical data of children with PAX2 gene mutation in Pediatric Nephrology Department, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from January 2014 to December 2022 were collected, and peripheral blood gene DNA was extracted and sequenced for whole exome sequencing. The clinical, pathological and genotypic characteristics of PAX2 gene variation of children in China were summarized by searching PubMed, Medline, China National Knowledge Infrastructure and Wanfang database and compared with the cases in this single center. Results:Among the 13 children with PAX2 gene mutation, there were 9 males and 4 females, 12 patients with abnormal urine tests, 7 patients with small kidney volume by imaging examination, and 5 patients with renal cysts. The clinical phenotypes were congenital renal and urinary tract malformations in 8 cases, renal coloboma syndrome in 1 case, and hematuria or proteinuria in 3 cases. Five patients underwent renal biopsies, showing focal segmental glomerulosclerosis and C3 glomerulopathy in 1 case, focal segmental glomerulosclerosis in 1 case, thin basement membrane lesion in 1 case, and IgA nephropathy in 2 cases. The genetic testing in 13 children showed 9 de novo mutations and 4 new mutations of c.321G>A, c.213-8C>G, c.63C>A and c.449C>T. There were 2 cases of 76dupG (p.V26Gfs*28) mutant. A total of 51 Chinese children with PAX2 gene mutation were found in the literature search. There were 32 males and 19 females, 8 cases with small kidney volume and 12 cases with renal cysts. The clinical phenotypes were congenital anomalies of kidney and urinary tract in 28 cases, renal coloboma syndrome in 17 cases, and hematuria or proteinuria in 6 cases. Seven patients underwent renal biopsies, including 2 cases with focal segmental glomerulosclerosis, 1 case with minimal lesion, 1 case with mesangial proliferative glomerulonephritis, 1 case with IgA nephropathy, 1 case with membranous nephropathy and a case with focal proliferative sclerosing purpura nephritis combined with glomerular hypertrophy. Thirty-four cases were de novo mutations, and 12 mutations were from the father or mother. The father or mother of 5 children had no clinical manifestations, with normal renal function. There were 11 cases of 76dupG (p.V26Gfs*28) mutant. Conclusions:The clinical phenotypes and genotypes of PAX2 gene variation in Chinese children are diverse. The most common clinical phenotype of PAX2 gene variation is congenital anomalies of kidney and urinary tract. c.76dupG (p.V26Gfs*28) is the most common of PAX2 gene variant.

Objective:To investigate the prognosis and risk factors of IgA vasculitis nephritis (IgAVN) in children.Methods:A retrospective cohort study was conducted. Clinical data were collected from 264 children who were pathologically diagnosed with IgAVN at Department of Pediatric Nephrology, Tongji Hospital, affiliated with Tongji Medical College, Huazhong University of Science and Technology, between January 2011 and December 2017. All patients had a follow-up period of more than 3 years. Clinical characteristics, renal pathology, 3-year and 5-year prognosis were analyzed. The patients were grouped based on gender, age of onset (≤6 years, >6-9 years, and >9 years), pathological classification (≤Ⅲ and>Ⅲ),whether the prognosis was complete remission at 3 and 5 years. Independent sample t-tests, ANOVA or chi-squared test were used for intergroup comparisons. Spearman correlation analysis was applied for ordinal data, and multivariate Logistic regression was used to analyze factors affecting the prognosis. Receiver operating characteristic (ROC) curve was utilized to evaluate the predictive value of these factors. Results:Of the 264 children with IgAVN, 153 were male and 111 were female, the age of onset was 8.3 (6.7, 10.3) years, 118 patients (45%) with onset age >6-9 years accounted for the highest proportion. All patients presented with skin purpura and renal involvement, primarily manifesting as hematuria and/or proteinuria. Microscopic hematuria was observed in 253 patients (95.8%), while 246 patients (93.2%) showed proteinuria. In 256 patients (97.0%), hematuria or proteinuria urinalysis was detected within 6 months of skin purpura onset, and 243 patients (92.0%) underwent renal biopsy within 6 months of renal involvement. The most common clinical subtype in 264 IgAVN children was hematuria and proteinuria (204 cases, 77.3%), with grade Ⅲ being the predominant pathological classification (181 cases, 68.6%). Among children ≤6 years old, the 3-year complete remission rate was higher in males than in females (83.9% (26/31) vs. 7/16, χ2=8.12, P=0.012). Factors independently associated with poor 5-year prognosis included time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission 3 years post-biopsy ( OR=5.41, 1.39, 6.02, 95% CI 1.40-20.86, 1.04-1.84, 2.61-13.88, all P<0.05). The serum cholesterol has a predictive value for 5-year prognosis ( P=0.020, AUC=0.62, 95% CI 0.52-0.71, Youden index=0.27, cutoff=4.37). Conclusions:For children with IgAVN aged≤6 years, the 3-year prognosis is better in males than in females. Time from hematuria or proteinuria urinalysis to renal biopsy >6 months, elevated serum cholesterol levels, and incomplete remission at 3 years post-biopsy may be independent risk factors for poor 5-year prognosis in children with IgAVN.

Objective To explore the effect and mechanism of berberine combined with curcumin on ath-erosclerosis(AS)by mediating M1 macrophages polarization.Methods M1-type macrophages were obtained from mouse mononuclear macrophages(RAW264.7)induced by lipopolysaccharide(LPS,100 ng/mL)and interferon(IFN)-γ(20 ng/mL).A cell model was established.The cells were divided into a control group,model group,berberine group,curcumin group and berberine plus curcumin group.Concentrations of berberine and curcumin were detected by CCK-8 assay.The expression levels of M1-type macrophage markers iNOS,TNF-α,CXCL9 and p-STAT6/STAT6 in macrophage supernatant were detected by ELISA.Levels of iNOS,TNF-α and CXCL9 mRNA were detected by RT-PCR.Levels of iNOS,STAT6 and p-STAT6 proteins in each group were detected by Western blot.After down-regulation of STAT6 level by siRNA technology,expression of p-STAT6 protein was detected by Western blot.Expression levels of iNOS,TNF-α,CXCL9 and p-STAT6 were detected by ELISA.Results In the polarization of M1 macrophages induced by LPS and IFN-γ,berberine(25 μmol/L)and curcumin(20 μmol/L)were the best concentrations as compared with other drug concentration groups,and neither alone nor combined use could significantly inhibit the viability of RAW264.7 cells(P<0.05).As compared with the normal group,iNOS,TNF-α and CXCL9 mRNA and protein levels were increased in the model group,while P-STAT6/STAT6 levels were decreased,with statistical differences(P<0.05).As compared with the model group,iNOS,TNF-α and CXCL9 mRNA and protein levels in the berberine group,curcumin group,and berberine plus curcumin group were decreased,while P-STAT6/STAT6 levels were increased,and the changes were more obvious in berberine plus curcumin group,with statistical difference(P<0.05).After transfection of STAT6 siRNA in M1 macrophages in the berberine plus curcumin group,P-STAT6 levels were down-regulated,while expressions of iNOS,TNF-α and CXCL9 were up-regulated,with statistical differences(P<0.05).Conclusions Both berberine and curcumin can inhibit the activity of M1-type macrophages and reduce inflammatory response.The action of berberine combined with curcumin is more advantageous than that of either drug alone,which may be the main mechanism of action through activation of STAT6.

Objective To compare the difference in the efficacies of arthroscopic microfracture operation for talar osteochondral injuries with hyperuricemia and non-hyperuricemia,and to explore the correlation be-tween blood urate level and efficacy.Methods Fifty-three patients with talar osteochondral lesions meeting the inclusion and exclusion criteria from February 2015 to August 2021 were selected as the research subjects and divided into the hyperuricemia group (22 cases) and non-hyperuricemia group (31 cases) according to whether or not the preoperative blood uric acid level exceeding 420 μmol/L.The joint range of motion (ROM),visual analog scale (VAS) score,American Foot and Ankle Surgery Society (AOFAS) score,mag-netic resonance score of cartilage repair tissue (MOCART) score and postoperative satisfaction before and af-ter surgery were compared between the two groups.Results The preoperative blood uric acid level in the hy-peruricemia group was higher than that in non-hyperuricemia group,and the difference was statistically signif-icant[(504.35±86.40)μmol/L vs.(332.56±45.80)μmol/L,P<0.05].The ROM score,VAS score and AOFAS score in postoperative 1 year follow up and last follow up in the two groups were significantly im-proved compared with before operation (P<0.001).The AOFAS scores before operation,in postoperative 1 year and postoperative last follow up in the hyperuricemia group were lower than those in the non-hyperurice-mia group (P<0.05).The VAS scores before operation and postoperative last follow up in the hyperuricemia group were higher than those in the non-hyperuricemia group (P<0.05).The uric acid level was negatively correlated with the postoperative AOFAS score (r2=0.076,P=0.041).The MOCART score in postopera-tive last follow up in the hyperuricemia group was lower than that in the non-hyperuricemia group,and the difference was statistically significant (P<0.05).The cartilage defect repair and filling degree and the fusion of repaired tissue with adjacent cartilage had statistical differences between the hyperuricemia group and non-hyperuricemia group (P<0.05).Conclusion Arthroscopic microfracture operation in treating talar osteo-chondral injuries has good clinical effect,the postoperative clinical effect in the patients with complicating hy-peruricemia is lower than that in the patients with non-hyperuricemia and the blood uric acid level is negative-ly correlated with the AOFAS score after microfracture surgery.

Objective To explore the effect and possible mechanism of berberine on the macro-phage polarization of human myeloid leukemia monocytic cell line THP-1 induced by oxidized low-density lipoprotein(ox-LDL).Methods THP-1 cells were induced into macrophages by PMA,and then according to different concentrations of berberine,the cells were divided into con-trol group,and 5,10,20,40 and 50 μmol/L berberine groups.After intervention for 24 or 48 h,CCK8 assay was used to detect cell viability for optimal concentration and time of berberine treat-ment.PMA-induced THP-1 macrophages were assigned into blank group,model group(ox-LDL),berberine group,inhibitor group(phosphatidyl inositol 3-kinase(PI3K)inhibitor LY294002)and berberine+inhibitor group(berberine+LY294002).The contents of inducible nitric oxide syn-thase(iNOS)and TGF-β1 were detected by ELISA.qPCR was employed to measure the mRNA expression of TNF-α,arginase 1(Arg1),PI3K and protein kinase B Akt1,and Western blotting was applied to detect the protein levels of Akt1 and phosphorylated protein kinase B antibody(p-Akt1).Results In 24 h after intervention,the macrophage activity was significantly lower in the 40 and 50 μmol/L berberine groups than the control group(P＜0.05),and after 48 h,the ac-tivity in all the 5 doses of berberine groups was obviously lower than that in the control group[(0.89±0.02)％,(0.82±0.03)％,(0.71±0.02)％,(0.62±0.03)％and(0.53±0.02)％vs(1.01±0.01)％,P＜0.05].Berberine treatment of 20 μmol/L for 24 h had little effect on cell viability,and the dose and the time were regarded as the best concentration and time.Compared with the blank group,iNOS content and TNF-α mRNA level were increased in the model group,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1,and p-Akt1/Akt1 protein levels were de-creased(P＜0.05).iNOS content and TNF-α mRNA level were decreased,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1s were increased in the berberine group than the model group(P＜0.05).Compared with the berberine group,iNOS con-tent and TNF-α mRNA level were increased,while mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1 were decreased in the berberine+inhibitor group(P＜0.05).Con-clusion Berberine can inhibit the inflammatory response of THP-1 macrophages induced by ox-LDL by activating PI3K/Akt1 pathway,and inhibit the M1 polarization and promote the M2 polarization of macrophages.