Objective: To explore the distribution of traditional Chinese medicine (TCM) syndromes in children with hepatolenticular degeneration (Wilson disease, WD) based on factor analysis and cluster analysis. Methods: From November 2018 to November 2023, general information (gender, age of admission, age of onset, course of disease, clinical staging, Western medicine clinical symptoms, and family history) and TCM four-examination informations (symptoms and signs) were retrospectively collected from 757 cases of children with WD at the First Affiliated Hospital of Anhui University of Chinese Medicine, and factor analysis and cluster analysis were used to investigate TCM syndromes in children with WD. Results: A total of 757 children with WD were included, of which 483 were male and 274 were female; the median age at admission was 12.58 years, the median age at onset was 8.33 years, and the median course of disease was 24.37 months; clinical typing result indicated 506 cases of hepatic type, 133 cases of brain type, 99 cases of mixed-type, and 19 cases of other type; 36.46% of the children had no clinical symptoms (elevated aminotransferases or abnormalities in copper biochemistry); a total of 177 cases had a definite family history, and 10 cases had a suspected family history. Forty-three TCM four-examination information were obtained, with the top 10 in descending order being feeling listless and weak, brown urine, slow action, inappetence, dim complexion, slurred speech, angular salivation, body weight loss, hand and foot tremors, and abdominal fullness. In children with WD, the syndrome element of disease location was primarily characterized by the liver, involving the spleen and kidney, and the syndrome elements of disease nature were characterized by dampness, heat, and yin deficiency. Based on factor analysis and cluster analysis, five TCM syndromes were derived, which were, in order, syndrome of dampness-heat accumulation (265 cases, 35.01%), syndrome of yin deficiency of the liver and kidney (202 cases, 26.68%), syndrome of liver hyperactivity with spleen deficiency (185 cases, 24.44%), syndrome of qi and blood deficiency (79 cases, 10.44%), and syndrome of yang deficiency of the spleen and kidney (26 cases, 3.43%). Conclusion The TCM syndromes of children with WD were primarily syndromes of dampness-heat accumulation, yin deficiency of the liver and kidney, and liver hyperactivity with spleen deficiency. The liver was the main disease location, and the disease nature was characterized by deficiency in origin and excess in superficiality, excess and deficiency mixed. These findings suggest that treating children with WD should be based on the liver while also considering the spleen and kidney.

Autism spectrum disorder in children (hereinafter called "childhood autism") is a prevalent neurodevelopmental condition; however, ancient Chinese medical texts did not provide detailed accounts of it. The concept of "the sovereign fire illuminates, while the ministerial fire occupies its position" was first introduced in the Plain Questions: The Great Treatise on the Origin of Heavenly Influences chapter. Subsequent medical practitioners have elaborated on this idea, identifying the sovereign fire as the master of the spirit and governing life activities, while the ministerial fire is the assisting minister that supports the functions of the zang-fu organs. When these two are in harmony, they complement each other, collectively sustaining the body′s normal physiological functions. In the context of this theory, the pathogenesis of childhood autism is attributed to the imbalance between the sovereign and ministerial fires. The root cause lies in the mutual antagonism of these two fires, resulting in the dysfunction of the spirit. Central to childhood autism onset is the dimming of the sovereign fire and a deficiency in the spirit, while the displacement of the ministerial fire and the impaired function of the five viscera contribute significantly to the progression of the condition. Effective treatment for childhood autism involves restoring these imbalances: dispelling yin-related obstructions to illuminate the sovereign fire, nourishing the spleen and kidney to nourish the ministerial fire, and regulating the liver and lung to harmonize the sovereign and ministerial fires. The ultimate goal is to integrate body and spirit, harmonize the sovereign and ministerial fires, and restore the interconnectedness between spirit and the function of the five zang viscera. This article, based on the "the sovereign fire illuminates, while the ministerial fire occupies its position" theory, provides a novel perspective and insights for understanding and treating childhood autism through the holistic principles of traditional Chinese medicine. Emphasizing the integration of body and spirit offers new insights and approaches for the clinical management of childhood autism in traditional Chinese medicine.

The lung collaterals form a network that branches from the lung meridian, traversing the lung system and extending across the body′s surface. Lung collateral disease refers to the structural alterations or dysfunction in these collaterals caused by external or internal pathogens. Research into the structural and physiological functions of children′s lung collaterals, as well as the pathogenesis and syndrome differentiation for treating lung collateral diseases in children, holds significant value in guiding the prevention and treatment of pediatric respiratory conditions. Drawing on the theory of collateral disease, the clinical insights of both historical and contemporary physicians, and modern research findings—while considering the unique physiological and pathological characteristics of children′s respiratory systems—this study provides a foundational summary of the morphology and spatial distribution of children′s lung collaterals. The characteristics of these collaterals are highlighted as thin, sparse, short, narrow, brittle, and tender. From this structural understanding, the unique physiological functions of children′s lung collaterals are analyzed. The study further explores the interactions between pathogenic factors and lung collaterals, elucidating the pathogenesis and progression of children′s lung collateral diseases. It proposes treatment principles centered on "seeking treatment in the collaterals and employing the method of unblocking collaterals, "which align with the unique features of pediatric lung collaterals. Common treatment approaches, and relevant prescriptions for managing these diseases are summarized. This paper lays the foundation for a theoretical system encompassing the structure, function, pathogenesis, and syndrome differentiation for treating children′s lung collateral diseases. It offers valuable insights for the clinical diagnosis and management of pediatric respiratory diseases linked to collateral dysfunction and serves as a reference for the systematic development of a broader theoretical framework for children′s collateral diseases.

Multiple system atrophy (MSA) is a rare neurodegenerative disease with complex clinical manifestations, presenting substantial challenges in clinical diagnosis and treatment. Its symptoms and the eight extraordinary meridians are potentially correlated; therefore, this article explores the association between MSA symptom clusters and the eight extraordinary meridians based on their circulation and physiological functions, as well as their treatment strategies. The progression from deficiency to damage in the eight extraordinary meridians aligns with the core pathogenesis of MSA, which is characterized by "the continuous accumulation of impacts from the vital qi deficiency leading to eventual damage". Liver and kidney deficiency and the emptiness of the eight extraordinary meridians are required for the onset of MSA; the stagnation of qi deficiency and the gradual damage to the eight extraordinary meridians are the key stages in the prolonged progression of MSA. The disease often begins with the involvement of the yin and yang qiao mai, governor vessel, thoroughfare vessel, and conception vessel before progressing to multiple meridian involvements, ultimately affecting all eight extraordinary meridians simultaneously. The treatment approach emphasizes that "the direct method may be used for joining battle, but indirect method will be needed in order to secure victory" and focuses on "eliminate pathogenic factors and reinforce healthy qi". Distinguishing the extraordinary meridians and focusing on the primary symptoms are pivotal to improving efficacy. Clinical treatment is aimed at the target, and tailored treatment based on careful clinical observation ensures precision in targeting the disease using the eight extraordinary meridians as the framework and core symptoms as the specific focus. Additionally, combining acupuncture, daoyin therapy, and other method may help prolong survival. This article classifies clinical manifestations based on the theory of the eight extraordinary meridians and explores treatment.

Objective : To pathological changes and inducible nitric oxide synthase(iNOS), phosphorylated insulin receptor substrate 1 serine 307(p-IRS1ser 307), phosphorylated protein kinase B serine 473(p-AKTser 473), glycogen synthase kinase-3β(GSK-3β), and gluconeogenic synthase(GS) proteins were observed in the liver of rats under the condition of chronic intermittent hypoxia-replicated oxygen in control. And to explore the role of iNOS/IRS1/AKT/GSK-3β signaling pathway in chronic intermittent hypoxia-induced insulin resistance. Methods : Forty SD rats were randomly divided into a control group(NC group) and an experimental group(CIH group), with 20 rats in each group. The NC group was placed in a normoxic environment for 12 weeks, while the CIH group was first subjected to intermittent hypoxia for 8 weeks, and then resumed normoxic rearing until the 12th week. Fasting blood glucose(FBG) and fasting insulin(FINS) were measured at baseline, week 8 and week 12, and liver tissues were taken for pathology and measurement of iNOS, p-IRS1ser 307, p-AKTser 473, GSK3β and GS levels, to compare the differences between groups. Results: t baseline, there was no significant difference in liver pathology between the two groups, and the observed indexes were not statistically significant(P>0.05); at 8 weeks, compared with the NC group, liver pathology in the CIH group showed significant disorganization of hepatic blood sinusoids and hepatocyte cords, obvious hepatocyte edema, smaller nuclei, increased lymphocyte infiltration, and a small number of fat vacuoles, significantly higher levels of FBG, FINS, insulin resistance index(HOMA-IR), iNOS mRNA, p-IRS1ser 307 protein, GSK-3β protein levels, and decreased p-AKTser 473 protein and GS protein levels, all of which were statistically significant(allP<0.05). IRS1ser 307 protein, GSK-3β protein levels were increased, p-AKTser 473 protein and GS protein levels were decreased, and the differences were statistically significant(allP<0.05); at 12 weeks, no lymphocyte infiltration was seen in the CIH group compared with that of the NC group and fat vacuoles significantly increased, and there was no improvement in the other pathological damage that had already occurred, and the levels of p-AKTser 473 protein significantly increased. AKTser 473 protein level significantly increased, p-IRS1ser 307 protein and GS protein levels were significantly reduced, all of which were statistically significant(allP<0.05), and the rest of the observational indexes were not statistically significant. Pearson&prime;s correlation analysis showed that HOMA-IR of CIH group was significantly positively correlated with the levels of iNOS mRNA, p-IRS1ser 307 protein, and GSK-3β protein at 8 weeks(r=0.874, 0.817,0.872;allP<0.05), and significantly negatively correlated with the levels of p-AKTser 473 protein and GS protein(r=-0.886,-0.879;allP<0.05). Conclusion Chronic intermittent hypoxia can lead to hepatic pathological damage that cannot be reversed even by reoxygenation interventions and may mediate the development of insulin resistance by upregulating the IRS1/AKT/GSK-3β signaling pathway through the upregulation of iNOS mRNA expression.

Objective : To explore the effect and mechanism of CD151 on vascular permeability by regulating vesicle internalization and recycling. Methods: Wild-type mice and CD151 knockout mice were divided into WT-con group, WT-model group, KO-con group and KO-model group, with 6 mice in each group. WT-model group and KO-model group were intraperitoneally injected with LPS to prepare sepsis ALI model, and WT-con group and KO-con group were intraperitoneally injected with phosphate buffer saline(PBS) as a control. 24 h after modeling, pulmonary vascular permeability was measured by Miles test. The siRNA silencing CD151 expression(si-CD151) and negative control si-NC were transfected into EA.hy 926 cells. The permeability of endothelial cell layer to FITC-dextran at different time points was observed under basic conditions and vascular endothelial growth factor-A(VEGF-A) stimulation conditions. Transcriptome sequencing of endothelial cells in si-CD151 group and si-NC group; the distribution and internalization of CD151 in each group were measured using immunofluorescence. Western blot and real-time quantitative RT-qPCR were used to detect the expression of VE-cadherin in si-CD151 groupand other groups. The distribution and internalization of VE-cadherin in each group were measured using immunofluorescence. Results : Miles experiment results indicated that dye exudation in lung tissue of WT-model group was significantly higher than that of WT-con group(P<0.01). The dye exudation in the lung tissue of KO-model group increased compared with WT-model group(P<0.05). The results of endothelial cell layer permeability test showed that the permeability of FITC-dextran in si-CD151 group was significantly higher than that in control group after VEGF-A stimulation for 30, 60 and 120 min(P<0.05). Transcriptome sequencing results suggested that CD151 in endothelial cells was closely related to vesicle-mediated transport. Compared with other groups, protein and mRNA levels of VE-cadherin in CD151 knockdown endothelial cells was significantly lower(allP<0.01). The immunofluorescence assay demonstrated that after VEGF-A stimulation, the decrease of CD151 expression significantly impaired the expression of VE-cadherin at cell-cell contacts and reduced the CD151-VE-cadherin colocalization in the perinuclear region compared with other groups. Conclusion The absence of CD151 affects the internalization and recycling of endothelial cell vesicles, affects the expression and internalization of VE-cadherin, and then influences vascular permeability.

Objective: To investigate whether syndecan-2(SDC2) can affect the proliferation, invasion and migration of cervical cancer cells by regulating ferroptosis and its possible mechanism. Methods : Normal cervical epithelial cells H8 and cervical squamous carcinoma cells C33A were cultured and divided into H8 group and C33A group. C33A cells were cultured and divided into control group, low SDC2 expression group, SDC2+ferroptosis inhibitor(ferrostation-1) group and SDC2 + ferroptosis inducer(erastin) group. Western blot was used to detect the protein levels of SDC2, solute carrier family 7 member 11(SLC7A11) and glutathione peroxidase 4(GPX4). RT-qPCR was used to detect the SDC2 mRNA level in C33A cells. ELISA kits were used to detect the levels of reactive oxygen species(ROS), glutathione(GSH) and ferrous ion(Fe2+) in C33A cells. The cloning ability of C33A cells was detected by plate cloning. The migration ability of C33A cells was detected by scratch test. Transwell assay was used to detect the invasion ability of C33A cells. Results : Compared with H8 group, the protein and mRNA expressions of SDC2, SLC7A11 and GPX4 in C33A group increased(P<0.05). Compared with the control group, the proliferation ability, migration ability and invasion ability of C33A cells in the low SDC2 group decreased(P<0.05), the protein and mRNA expressions of SLC7A11 and GPX4 in C33A cells decreased(P<0.05), and the GSH level decreased. ROS and Fe2+levels increased(P<0.05). Compared with the low SDC2 group, the protein and mRNA expressions of SLC7A11 and GPX4 increased(P<0.05), the GSH level increased, and the ROS and Fe2+levels decreased(P<0.05) in the low SDC2+ferrostation-1 group. Compared with the control group, the proliferation ability, migration ability and invasion ability of C33A cells with low SDC2+erastin expression decreased(P<0.05). Conclusion The expression of SDC2 increases in C33A cervical cancer cells. Low expression of SDC2 can activate SLC7A11/GPX4 pathway mediated ferroptosis, thereby reducing the proliferation, invasion and migration of C33A cells.

Objective : To explore the effect of dendrobium officinale polysaccharides(DOP) on necrotizing enterocolitis(NEC) in neonatal mice and to preliminarily explore the potential molecular mechanisms. Methods : Seven-day-old C57BL/6J mice were randomly divided into four groups: control(CTRL) group, necrotizing enterocolitis(NEC) group, low-dose DOP treatment(DOPL+NEC) group and high-dose DOP treatment(DOPH+NEC) group. The NEC model was established by hypoxia, cold stimulation, hypertonic feeding and intraperitoneal injection of lipopolysaccharide(LPS). At the end of the experiment, the small intestine tissues were collected. The general condition of the mice was observed, and body weight was recorded. Hematoxylin-eosin(HE) staining was used to observe pathological changes in the small intestine tissues. The expression levels of E-cadherin, Occludin and Claudin-1 were detected by immunohistochemistry and Western blot. The concentrations of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, IL-6 and IL-10 were measured by ELISA. The levels of lactate dehydrogenase(LDH), malondialdehyde(MDA), superoxide dismutase(SOD) and glutathione(GSH) were detected using commercial kits. The expression levels of nuclear factor erythroid 2-related factor 2(Nrf2) and heme oxygenase-1(HO-1) proteins were detected by immunohistochemistry and Western blot. Results : ompared with the CTRL group , the NEC group exhibited decreased body weight and increased HE pathological scores (P < 0. 01 or P < 0. 001) . The protein expression levels of E-cadherin , Occludin and Claudin-1 were reduced (P < 0. 01 or P < 0. 001) . The concentrations of TNF-α, IL-1βand IL-6 increased , while the concentration of IL-10 decreased (P < 0. 01) . The levels of MDA and LDH increased , while the levels of GSH and SOD decreased (P < 0. 01) . The protein expression levels of Nrf2 and HO-1 increased ( P < 0. 05 or P < 0. 01) . In contrast , DOP intervention groups showed increased body weight and decreased HE scores (P < 0. 05 or P < 0. 01) . The protein expression levels of E-cadherin , Occludin and Claudin-1 increased (P < 0. 05 or P < 0. 01 or P < 0. 01) . The concentrations of TNF-α, IL-1βand IL-6 decreased(P < 0. 05 or P < 0. 01 ) , while the concentration of IL-10 increased ( P < 0. 01) . The levels of MDA and LDH decreased , while the levels of GSH and SOD increased ( P < 0. 05 or P < 0. 01) . The protein expression levels of Nrf2 and HO-1 further increased (P < 0. 05 or P < 0. 01) . Conclusion DOP can ameliorate the pathological damage of necrotizing enterocolitis and enhance the intestinal mucosal barrier function in NEC mice . Additionally , it can also reduce oxidative stress injury and intestinal inflammation in NEC mice . The mechanism may be associated with the activation of the Nrf2/HO-1 pathway by DOP.

Objective : To investigate the role and mechanism of long non-coding RNA(lncRNA) H19 in osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs) and bone regeneration in mice with foot fracture. Methods: BMSCs were divided into microrNA-149-5 p(miR-149-5 p)-mimic negative control(miR-mimicNC) group and miR-149-5p-mimic(miR-mimic) group and transfected with IncRNA H19-wt and IncRNA H19,respectively.Mut recombinant plasmid,or bone morphogenetic protein 2(BMP2) 3'UTR-wt and BMP2 3' UTRmut recombinant plasmid were transfected,respectively.Dual-luciferase reporter gene assay was used to detect the luciferase activity of each group.To study the regulatory effect and mechanism of IncRNA H19 on osteogenic differentiation in vitro,BMSCs were divided into control group and osteogenic induction group(osteogenic group).After osteogenic induction,BMSCs were transfected with corresponding plasmids and divided into osteogenic+si-NC group,osteogenic+si-H19 group,osteogenic+si-H19+miR-inhibitor-NC group,osteogenic+si-H19+miR-inhibitor group,osteogenic+si-H19+pcDNA-NC group,and osteogenic group si-H19+pcDNA-BMP2 group.Osteogenic differentiation was evaluated by alkaline phosphatase(ALP) activity assay and alizarin red staining.A mouse foot fracture model was established,and 36 mice were randomly divided into sham operation group,model group,model+pcD-null group,and model+pcD-H19 group,with 9 mice in each group.Osteogenic differentiation was assessed by ALP activity assay.Real-time fluorescent quantitative PCR(qPCR) was used to detect the expression of IncRNA H19,miR-149-5p,and BMP2.Western blot was used to detect the expression of BMP2,OCN,OSX,RUNX2,and OPN. Results : In cells transfected with lncRNA H19-wt , the luciferase activity in miR-mimic group was lower than that in miR-mimic-NC group (P < 0. 05) . The luciferase activity of miR-mimic group was lower than that of miR-mimic-NC group ( P < 0. 05) . Compared with the control group , the alkaline phosphatase activity , the degree of cell mineralization and the expression of lncRNA H19 , BMP2 , OCN , OSX , RUNX2 and OPN increased , and the expression of miR-149-5p decreased in the osteogenic group ( P < 0. 05) . Compared with the osteogenesis + si-NC group , the alkaline phosphatase activity , the degree of cell mineralization and the expression of lncRNA H19 , BMP2 , OCN , OSX , RUNX2 and OPN significantly decreased , and the ex- pression of miR-149-5p increased in the osteogenesis + si-H19 group (P < 0. 05) . Compared with the osteogenesis + si-H19 + miR-inhibitor-NC group , the ALP activity , the degree of cell mineralization and the expression of ln- cRNA H19 , BMP2 , OCN , OSX , RUNX2 and OPN increased in the osteogenesis + si-H19 + miR-inhibitor group . The expression of miR-149-5p significantly decreased (P < 0. 05) . Compared with the osteogenic + si-H19 + pcD- NA-NC group , the alkaline phosphatase activity , the degree of cell mineralization and the expression of BMP2 , OCN , OSX , RUNX2 and OPN significantly increased in the osteogenic + si-H19 + pcDNA-BMP2 group ( P < 0. 05) . Compared with the sham-operation group and the model group , the alkaline phosphatase activity and the expression of lncRNA H19 , BMP2 , OCN , OSX , RUNX2 and OPN decreased , and the expression of miR-149-5p increased (P < 0. 05) . Compared with the model + pcD-null group , the alkaline phosphatase activity and the ex- pression of lncRNA H19 , BMP2 , OCN , OSX , RUNX2 and OPN significantly increased , and the expression of miR-149-5p decreased in the model + pcD-H19 group (P < 0. 05) . Conclusion lncRNA H19 promotes osteogenic differentiation of BMSC s and bone regeneration in mice with foot fracture through miR-149-5p/BMP2 axis .

Objective : To investigate the role of endoplasmic reticulum stress in the occurrence and development of fatty liver induced by high fat. Methods : In the high-fat Drosophila model, the high-fat group was fed with high-fat medium, while the control group was fed with normal medium; in the mouse fatty liver model, the high-fat group was fed with high-fat diet, and the control group was fed with normal diet; in the HepG2 cell steatosis model, the high-fat group was induced by palmitic acid(PA), and the control group was cultured with DMEM. The fat body size of the third instar larvae of Drosophila melanogaster was photographed. Steatosis in mice liver and HepG2 cells was observed by H&E and Oil Red staining. The expression levels of ATF6, Bip and CHOP in the third instar larvae, liver tissues of mice and HepG2 cells were analyzed by quantitative real-time polymerase chain reaction(qPCR) and Western blot. Results : In Drosophila model, fat body and fat storage were obviously increased in high fat fed flies when compared with control group. The formation of liver fat droplets and cells vacuolation were confirmed by H&E and Oil Red staining in mice livers fed with high fat and HepG2 cells with palmitic acid treatment. The expression levels of ATF6, Bip and CHOP were significantly increased in third instar larvae and mice livers fed with high fat and palmitic acid treated HepG2 cells with palmitic acid treatment. Conclusion High fat may induce the occurrence and development of hepatic steatosis by activating endoplasmic reticulum stress.