Attention deficit hyperactivity disorder （ADHD） is often accompanied by tic disorder. The core pathogenesis is considered to be ministerial fire scorching yin and internal stirring of deficient wind， which leads to disharmony between the body and spirit， resulting in clinical manifestations. The treatment principles emphasize nourishing yin fluids， calming ministerial fire， and extinguishing endogenous wind （内风）. The method of nourishing yin fluids is applied throughout the entire treatment process， commonly using ingredients such as Shudihuang （Rehmanniae Radix Praeparata）， Shanzhuyu （Corni Fructus）， Gouqizi （Lycii Fructus）， Wuweizi （Schisandrae Chinensis Fructus）， and Tusizi （Cuscutae Semen）. These are combined with approaches to harmonize the zang-fu organs， primarily including extinguishing liver wind， clearing heart fire， nourishing kidney water， and strengthening spleen earth， thereby stabilizing ministerial fire and extinguishing endogenous wind. Additionally， emotional regulation and smoothing emotional constraint are essential to improve clinical symptoms in children with ADHD comorbid with tic disorder.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

ObjectiveZheng’s San Qi San (ZSQS) power, a classic traditional Chinese medicine (TCM) formula, is used for treating soft tissue injuries involving muscles, tendons, and ligaments. However, its underlying therapeutic mechanisms remain unclear. This study aimed to screen and identify pharmaceutically active ingredients and their candidate biomolecule targets, and further elucidate the molecular mechanism of ZSQS in the treatment of skeletal muscle injury. MethodsNetwork pharmacology was employed to construct “ZSQS-component-target”, “protein-protein interaction (PPI)” and “active ingredient-core protein-pathway” networks to predict the key active ingredients and potential core targets of ZSQS for skeletal muscle injury. The predicted results were then validated via microarray data from the GEO database. Molecular docking was then performed to assess the binding ability between the screened active ingredients of ZSQS and the candidate core targets. Moreover, liquid chromatography-mass spectrometry (LC-MS) was used for qualitative and quantitative analysis to verify the active components of the drug and ZSQS serum. Finally, an animal model of eccentric exercise-induced skeletal muscle injury and a myotube cell model of oxidative stress-induced injury were established to validate the effects of ZSQS and its interventional effects on the biological functions of critical targets, thereby demonstrating the potential therapeutic mechanism of ZSQS. ResultsAmong the 111 active components identified in ZSQS and their corresponding 204 targets related to the skeletal muscle injury repair process, 14 core targets (including AKT1) and 4 core active components (quercetin, luteolin, kaempferol, and β‑sitosterol) were screened out, while the corresponding metabolites of quercetin, luteolin and kaempferol were detected in the ZSQS serum. Among these targets, 5 candidate genes (IL-6, CASP3, HIF1A, STAT3, and JUN) overlapped with the differential expression screening results with GEO data, and IL-6 was confirmed to be enriched in the PI3K/AKT pathway. Combined with the prediction results of the AKT expression levels, these findings suggest that the phosphorylation level of AKT1 plays a core role in the therapeutic mechanism of ZSQS. Molecular docking analysis further revealed that the PH domain of AKT1 had high binding energy with all 4 core active components, as verified by LC-MS. Finally, animal model studies have shown the promoting effect of ZSQS administration on skeletal muscle injury repair and its possible antioxidant damage mechanism. Cell model studies further demonstrated that ZSQS-containing serum, core active ingredient combination therapy, and quercetin monomer could increase the phosphorylation level of AKT, promote the nuclear translocation of Nrf2, upregulate the expression of downstream antioxidant enzymes (SOD, GPx, and GR), and inhibit the expression of inflammatory factors (IL-6 and TNF-α), thereby alleviating oxidative stress and the inflammatory response. ConclusionZSQS alleviates skeletal muscle injury mainly by activating the AKT/Nrf2 signaling pathway, enhancing cellular antioxidant and anti-inflammatory capabilities. The results of this study provide a scientific basis for the clinical application and modernized development of ZSQS.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

This study aimed to establish an analytical method capable of simultaneously quantifying the prototype components of Si-Ni-San and screening their metabolites to elucidate its tissue distribution and metabolic characteristics in an ulcerative colitis (UC) mouse model. To this end, an integrated analysis strategy based on UPLC-QqQ-MS was developed and validated, combining the targeted quantification of 12 prototype components with a pseudo-targeted metabolite screening technique based on ion pair list-triggered data-dependent acquisition. Samples were extracted with 80% methanol (100 mg/mL) and processed with internal standards. Separation was achieved on a Waters Acquity UPLC HSS PFP column using a gradient elution with a mobile phase consisting of acetonitrile and 5 mmol/L ammonium acetate containing 0.1% formic acid. Analysis was performed using an electrospray ionization (ESI) source in multiple reaction monitoring mode. Methodological validation confirmed that all parameters met the requirements for bio-sample analysis. Application of this method revealed that the content of Si-Ni-San prototype components in the colon, liver, and kidneys of UC mice was significantly higher than that in healthy mice. Furthermore, the number of phase II metabolites was markedly greater than that of phase I metabolites in all tested samples. The results demonstrate the reliability of the established method and preliminarily reveal the tissue distribution characteristics of Si-Ni-San under UC conditions and its metabolism pattern dominated by phase II conjugation, which provides a methodological basis and experimental data for further in-depth research into its effective target tissues and pharmacodynamic material basis.

Objective: To investigate the trend of diabetes incidence among children and adolescents in Huzhou City, Zhejiang Province from 2006 to 2025, and to analyze the age, period, and cohort effects, so as to provide insights into the prevention and control of diabetes among children and adolescents. Methods: Data on diabetes cases aged 0-19 years in Huzhou City from 2006 to 2025 were collected through the Chronic Disease Surveillance and Management System of Huzhou Center for Disease Control and Prevention. The standardized incidence was calculated using the age composition of the standard population from the Sixth National Population Census in 2010. The average annual percentage change (AAPC) was used to analyze the trend in incidence. An age-period-cohort model was used to analyze the effects of age, period, and birth cohort. Results: A total of 913 diabetes cases aged 0-19 years were reported in Huzhou City from 2006 to 2025. The crude incidence and standardized incidence were 11.03/105 and 4.33/105, respectively, both showing upward trends (AAPC=13.307% and 13.226%, both P<0.05). There were 339 cases of type 1 diabetes mellitus (T1DM), with crude and standardized incidence of 3.83/105 and 0.62/105, respectively, showing upward trends (AAPC=9.944% and 10.442%, both P<0.05). There were 548 cases of type 2 diabetes mellitus (T2DM), with crude and standardized incidence of 6.20/105 and 1.97/105, respectively, showing upward trends (AAPC=14.920% and 14.213%, both P<0.05). Age-period-cohort model analysis showed that the risk of T1DM initially increased and then decreased with age, with the highest risk in the 5-<10 years age group (RR=9.189, 95%CI: 4.871-13.684). The risk of T2DM increased with age, with the highest risk in the 15-19 years age group (RR=17.316, 95%CI: 14.477-20.225). Using the period 2011-2015 as the reference group, the risk of T1DM showed an upward trend over time, with the highest risk in 2021-2025 (RR=1.474, 95%CI: 1.027-2.114). There were no statistically significant differences in incidence risk across other periods compared to 2011-2015 (all P>0.05). Using the 2006-2010 birth cohort as the reference group, the risks of both T1DM and T2DM showed upward trends with later birth cohorts. The highest risk of T1DM was observed in the 2021-2025 birth cohort (RR=2.881, 95%CI: 1.528-5.429), while the highest risk of T2DM was observed in the 2011-2016 birth cohort (RR=1.610, 95%CI: 1.081-2.397). Conclusions The incidence of diabetes among children and adolescents in Huzhou City showed an upward trend from 2006 to 2025, with T1DM being most prevalent among children aged 5-<10 years and T2DM among adolescents aged 15-19 years. The incidence risks of both T1DM and T2DM increased with later birth years.

Purpose: Cancer has become a significant major public health concern, making the discovery of new cancer markers or therapeutic targets exceptionally important. Elevated expression of tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) expression has been observed in certain types of cancer. This project aims to investigate the function of TNFRSF12A in tumors and the underlying mechanisms. Materials and Methods: Various websites were utilized for conducting the bioinformatics analysis. Tumor cell lines with stable knockdown or overexpression of TNFRSF12A were established for cell phenotyping experiments and subcutaneous tumorigenesis in BALB/c mice. RNA-seq was employed to investigate the mechanism of TNFRSF12A. Results: TNFRSF12A was upregulated in the majority of cancers and associated with a poor prognosis. Knockdown TNFRSF12A hindered the colorectal cancer progression, while overexpression facilitated malignancy both in vitro and in vivo. TNFRSF12A overexpression led to increased nuclear factor кB (NF-κB) signaling and significant upregulation of baculoviral IAP repeat containing 3 (BIRC3), a transcription target of the NF-κB member RELA, and it was experimentally confirmed to be a critical downstream factor of TNFRSF12A. Therefore, we speculated the existence of a TNFRSF12A/RELA/BIRC3 regulatory axis in colorectal cancer. Conclusion TNFRSF12A is upregulated in various cancer types and associated with a poor prognosis. In colorectal cancer, elevated TNFRSF12A expression promotes tumor growth, potentially through the TNFRSF12A/RELA/BIRC3 regulatory axis.

The concept of harmony between heaven and man is an important concept in TCM. The theory of "harmful hyperactivity and responding inhibition" has been developed and perfected by medical professionals throughout history to explain physiological and pathological changes in humans based on a natural understanding. Guided by the TCM philosophy of "harmful hyperactivity and responding inhibition", it provides guidance for attention deficit hyperactivity disorder (ADHD). Sorting out the complex pathogenesis, it has been found that core pathogenesis of the disease is the excessive damage to the five organs, which leads to the overall state of yin deficiency and yang hyperactivity in the human body. The treatment mainly focuses on regulating the hyperactivity and damage of the organs, restoring the balance of yin and yang, while also taking into account the harm of pathological products. Based on clinical manifestations, the secondary selection of medication for liver and liver hyperactivity and spleen and kidney dysfunction has been determined, while improving the hyperactivity of the external environment, in order to achieve the goal of "tolerance is control", which fully embodies the TCM syndrome differentiation thinking of "unity of heaven and man, holistic concept".

Objective To identify the impact of glucose variability(GV)on the development of gestational hyper-tension(GH)in patients with gestational diabetes mellitus(GDM),and to find the differences between various GV metrics as well as to evaluate their predictive value in management strategy development.Methods A total of 127 pregnant women diagnosed with GDM were included in this study.After the diagnosis of GDM,continuous glu-cose monitoring(CGM)and blood pressure measurements were performed,and the occurrence of gestational hyper-tensive disorders was recorded.Indices of glucose variability were calculated using an automated software EasyGV version 9.0.Results The results revealed an association between gestational hypertension and glucose variability in GDM patients.Among the study participants,2 cases(1.6％)were diagnosed with preeclampsia and 9 cases(7.1％)were diagnosed with gestational hypertension.TBR％(time below range)showed a significantly negative correlation with diastolic blood pressure at 29-32 weeks of gestation and with both diastolic and systolic blood pres-sure during delivery.TIR％(time in range)showed a negative correlation with the rate of change in systolic blood pressure between two prenatal visits.CONGA(continuous overlapping net glycemic action)emerged as an inde-pendent predictor of gestational hypertension(OR:3.648;95％CI:1.046,12.721;P=0.042).When CONGA exceeded 4.856,the risk of gestational hypertension in GDM patients increased.Conclusions Blood glucose varia-tion is an independent factor affecting the occurrence of pregnancy-induced hypertension in GDM women.This study suggests new targets for the use and management of CGM in pregnant women with GDM.For GDM patients at high risk of hypertensive disease during pregnancy,blood glucose should be kept at a reasonable and stable level.