This paper summarizes Professor LIU Fengbin's clinical experience in treating autoimmune liver disease （AILD） using the method of nourishing yin and removing stasis based on stage differentiation. He believes that the pathogenesis of AILD generally involves both deficiency in essence and excess in manifestation， with essence deficiency often presenting as liver and kidney yin deficiency， which may progress to spleen deficiency and yang deficiency over time. The excess manifestation commonly includes qi stagnation， blood stasis， damp-heat， and phlegm toxicity. Clinically， he advocates for the treatment principle of nourishing yin and removing stasis. On the foundation of nourishing liver and kidney yin， different pathological factors causing stasis are eliminated according to their nature. Treatment is also tailored to different stages of AILD. In the early and asymptomatic stages， liver qi stagnation and spleen deficiency are prominent， warranting a therapeutic approach of soothing the liver， regulating qi and strengthening the spleen. The modified Chaishao Qizhi Decoction （柴芍气滞汤） is used. During the symptomatic stage， pathogenic factors become more pronounced， often accompanied by a significant deficiency of vital qi， with damp-heat， water retention， and phlegm toxicity as key pathological features. The treatment should focus on strengthening the spleen and dispelling dampness， using modified Sijunzi Decoction （四君子汤） combined with Yinchen Wuling Powder （茵陈五苓散）. In the liver function decompensation stage， vital qi is severely deficient while pathogenic factors persist， with damp-heat， phlegm toxicity， and blood stasis obstructing the liver collaterals. Treatment should focus on nourishing blood， softening the liver， strengthening the spleen， and resolving stasis， using the modified Ruangan Yangxue Decoction （软肝养血汤）. Throughout the treatment process， emphasis is placed on tonifying the liver and kidneys while protecting yin fluids.

OBJECTIVE To explore the effect and mechanism of Zexie tang （ZXT） on reducing lipid accumulation through network pharmacology， and compare the difference of traditional decoction versus formula granules. METHODS The active components and targets of ZXT were identified using TCMSP and SwissTargetPrediction databases. GeneCards， OMIM， DisGeNET and TTD databases were used to analyze the related targets of non-alcoholic fatty liver disease （NAFLD）； protein-protein interaction network model was constructed by String database；“ ZXT-NAFLD target-pathway” network diagram was constructed by using CytoScape software； target enrichment analysis was performed by using Metascape platform. Fat accumulation model of human hepatocellular carcinoma HepG2 cells was established to observe the effects of traditional decoction and formula granules of ZXT on lipid accumulation of cells. RESULTS Alisol B， alisol C， 1-monolinolein and alisol B monoacetate were the key active components of ZXT in the treatment of NAFLD. The core targets included MDM2， MAPK1， PIK3CB， PRKCQ and MAPK14， etc. The core signaling pathways included endocrine resistance， insulin resistance and Th17 cell differentiation. Compared with model group， except for the Zexie formula granules group and Baizhu formula granules group， the absorbance values in all other administration groups were significantly decreased （P＜0.01）； the absorbance value of Baizhu traditional decoction group was significantly higher than that of ZXT traditional decoction group （P＜0.01）； the absorbance values of Zexie formula granule group and Baizhu formula granule group were significantly higher than that of ZXT formula granule group （P＜0.01）； the absorbance value of Zexie formula granule group was significantly higher than that of Zexie traditional decoction group （P＜0.01）； the absorbance value of Baizhu formula granule group was significantly higher than that of Baizhu traditional decoction group （P＜0.01）. CONCLUSIONS ZXT reduces lipid accumulation of human hepatocellular carcinoma cells through multiple components， multiple target and multiple pathways， and its traditional decoction and formula granules exhibit slightly different lipid-lowering effects.

Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine.

In the field of dental aesthetics,digital aesthetic design plays a crucial role in helping dentists to predict treatment outcomes vis-ually,as well as in enhancing the consistency of knowledge and understanding of aesthetic goals between dentists and patients.It serves as the foundation for achieving ideal aesthetic effects.However,there is no clear standard for this digital process currently in China and abroad.Many dentists lack of systematic understanding of how to carry out digital aesthetic design for treatment.To establish standardized processes for dental aesthetic design and to improve the homogeneity of treatment outcomes,Chinese Society of Digital Dental Industry(CSD-DI)convened domestic experts in related field to compile this consensus.This article elaborates on the key aspects of digital aesthetic data collection,integration steps,and the digital aesthetic design process.It also formulates a decision tree for dental aesthetics at macro level and outlines corresponding workflows for various clinical scenarios,serving as a reference for clinicians.

Objective To explore the efficacy and safety of enhanced external counterpulsation(EECP)in elderly patients with acute ischemic stroke(AIS)complicated by coronary heart dis-ease(CHD).Methods A total of 65 AIS patients with CHD admitted in our hospital from Janu-ary to June 2023 were recruited and randomly divided into a control group(drug secondary pre-vention,n=32)and a treatment group(drug combined with EECP therapy,n=33).Their NIHSS score,mRS score and Canadian Cardiovascular Society(CCS)angina grade were evaluated before and after treatment and compared between the two groups.The incidences of recurrent ischemic stroke,new hemorrhagic stroke and major adverse cardiovascular events(MACE)were also recor-ded during treatment.Results The NIHSS score and mRS score were significantly decreased in both groups after treatment(P＜0.01).After treatment,the NIHSS score(2.67±1.63 vs 3.56± 1.83),mRS score[1.0(0.0,1.0)vs 2.0(1.0,2.0)]and CCS grade[1.0(1.0,2.0)vs 2.0(1.0,2.0)]were obviously lower in the treatment group than the control group(P＜0.05,P＜0.01).There were no statistical differences in the incidence rates of recurrent ischemic stroke,new-onset hem-orrhagic stroke,and MACE between the control group and the treatment group(9.4％vs 6.1％,6.3％vs 3.0％,12.5％vs 6.1％,P＞0.0 5).Conclusion EECP is a safe and effective treatment option for elderly AIS patients with CHD.

【Objective】 To understand factors associated with children′s attendance at nursery care institutions (NCIs) and the basic characteristics of the NCIs children were enrolled in, so as to provide scientific evidence for policymakers. 【Methods】 During November 2020 and January 2021, parents who visited the Department of Child Health Care in six selected maternal and children′s hospitals, as well as nearby NCIs in Zhejiang were invited to finish an online questionnaire. Information such as children′s sociodemographic characteristics, parents′ knowledge, attitude and practice regarding nursery and feeding, etc. were collected. A total of 1 756 questionnaires were collected. 【Results】 Compared to children who were not in NCIs, children enrolled in NCIs were older (94.4% of children ≥24 months vs. 30.1%, χ²=835.27), more likely to be from the local area (87.2% vs. 81.4%,χ²=12.25), more likely to have parents with a college degree (mother: 83.6% vs. 74.2%, χ²=35.29; father: 79.9% vs. 70.0%, χ²=27.01), had a higher prevalence of family annual income >200 000 CNY (49.5% vs. 28.2%, χ²=110.49), and were less likely to have their grandparents available to take care of them (16.7% vs. 26.8%, χ²=31.4) The difference all have great significant.(P<0.05). In a multivariate Logistic regression model, the older the child, the more likely they were to attend an NCI (for children aged 6 - 23 months, OR=6.70; for children aged 24 - 35 months, OR=134.03; and for children aged 36 - 42 months, OR=699.33; P<0.05). Family annual income was positively associated with children′s attendance at NCIs (for those earning 100 000 - 200 000 CNY/year, OR=1.63; for those earning 200 000 - 500 000 CNY/year, OR=2.96; and for those earning >500 000 CNY/year, OR=4.62, P<0.05). Conversely, the higher the level of grandparent involvement in daily care, the lower the attendance at NCIs (for children cared for by both parents and grandparents, OR=0.57; for those primarily cared for by grandparents, OR=0.26, P<0.05). For children who used to stay at NCIs, 82.8% stayed at institutions that only recruited children aged 0 - 3 years, 97.4% spent their whole day in NCIs, and 71.4% spent less than 3 000 CNY per month for NCI services. Additionally, over 95% of parents were satisfied with the food and care services in NCIs, as well as their children′s physical development in NCIs. However, 32.1% of NCIs were reported by parents as having no room for breastfeeding. 【Conclusions】 Children′s age, grandparent involvement in routine care, and family annual income are the main factors associated with children′s attendance at NCIs. There is a greater need for more affordable and community-based NCIs, particularly for children under 2 years old. Additionally, more attention should be paid to the quality surveillance, assessment and management of NCIs.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Objective The purpose of this study was to explore a suitable method to model no-reflow phenomenon following ischemic stroke and to evaluate perfusion decrease from multiple perspectives.Methods Laser scatter contrast imaging and two-photon live imaging were used to compare transient middle cerebral artery occlusion in C57BL/6 and BALB/c mice and perfusion alterations in BALB/c mice with 1 or 1.5 h of ischemia.Several imaging techniques including laser scatter contrast imaging,low and higher magnification images of perfused brain slices and two-photon microscopy to monitor erythrocyte flow rate and flux were used to assess in vivo dynamics as well as whole brain sections and microvasculature for decreased cerebral perfusion after transient middle cerebral artery occlusion.Infarct size and behavioral deficits were assessed with microtubule-associated protein 2 staining and behavioral scoring.Results In C57BL/6 mice,most capillaries in the middle cerebral artery region remained flowing during ischemia,whereas most capillaries were blocked in BALB/c mice.In addition,cortical perfusion at 24 h of recanalization was significantly reduced to 76.1%of baseline following 1.5 h of ischemia in BALB/c mice(P=0.046 compared with the sham group),whereas for it was reduced to 79.9%following 1h of ischemia which was not significantly different from the sham group(P=0.299).Transient middle cerebral artery occlusion in BALB/c mice for 1.5 h resulted in a reduction in whole-brain perfusion to 75.1%(P＜0.001 compared with the sham group),and erythrocyte flow rate assessed by two-photon live-imaging of erythrocyte flow on the cortical surface of the middle cerebral artery basin was reduced to 50.3%of baseline levels at 24 h of recanalization(P=0.010 compared with the sham group),and erythrocyte flux decreased to 38.9%of baseline levels(P= 0.010 compared with the sham group);high-magnification imaging of sections assessed an approximately 76%reduction in the length of capillaries with perfusion(P=0.0001 compared with the sham group),and a reduction in the fraction of the total volume occupied by perfused capillaries by an approximately 76%reduction(P＜0.001 compared with the sham-operated group).Microtubule-associated protein 2 staining suggested that transient middle cerebral artery occlusion for 1.5 h in BALB/c mice resulted in infarcts that accounted for approximately 36%of the total cerebral area and behavioral scores elevated to 9,suggesting behavioral deficits.Conclusion Transient ischemia in BALB/c mice for 1.5 h resulted in a significant decrease in cerebral perfusion as well as capillary no-reflow and thus can model the no-reflow phenomenon following ischemic stroke.The combination of laser scatter contrast imaging,low magnification and higher magnification images of perfused brain slices,and two-photon microscopy live imaging allows for a multifaceted assessment of perfusion changes.

The complex pathophysiological mechanisms between diabetes mellitus and cardiovascular diseases have not yet been fully elucidated， becoming one of the challenges in clinical care. Glucagon-like peptide-1 receptor agonist （GLP1-RA） and sodium glucose cotransporter-2 inhibitors （SGLT2） are clinically used to reduce the cardiovascular risk of patients with diabetes mellitus. Traditional Chinese medicine has diverse biological activities and unique advantages in the treatment of chronic complex diseases due to its multi-component and multi-target effects. Based on recent reports， this paper reviewed the common risk factors of diabetes mellitus and cardiovascular diseases （e.g.， hyperglycemia， insulin resistance， and inflammation）， related targets such as apolipoprotein C-Ⅲ （APOC3）， S100 calcium-binding protein A8/A9 （S100A8/A9）， growth/differentiation factor-15 （GDF-15）， and NACHT， LRR， and PYD domains-containing protein 3 （NLRP3）， advanced glycation end products， insulin resistance， endothelial dysfunction， endoplasmic reticulum stress， mitochondrial dysfunction， and intestinal flora disorder. In addition， this paper summarized the research progress in the treatment of cardiovascular diseases in diabetes mellitus with the active ingredients （e.g.， baicalein， puerarin， curcumin， notoginsenoside， and tanshinone Ⅱ_A）， single herbal medicines （e.g.， Astragali Radix， Ginseng Radix et Rhizoma， Sophorae Flavescentis Radix， Cinnamomi Cortex， and Corni Fructus）， and compound formulas （e.g.， Buzang Tongluo Fang， Yiqi Yangyin Huoxue Fang， Shenqi Fang， Huangqisan， Danggui Buxue Tang， and Liuwei Dihuang Wan） of traditional Chinese medicine. Traditional Chinese medicine mainly treats cardiovascular diseases in diabetes mellitus by reducing inflammation and oxidative stress， ameliorating dyslipidemia and insulin resistance， protecting islet β cell function， repairing endothelial damage， inhibiting smooth muscle cell proliferation， foam cell formation， macrophage polarization， and cardiac hypertrophy and fibrosis， and regulating intestinal flora disorder. These processes involve insulin receptor substrate/ phosphatidylinositol 3-kinase/protein kinase B （IRS/PI3K/Akt）， peroxisome proliferator-activated receptor α/γ （PPAR α/γ）， nuclear factor-kappa B （NF-κB）， adenosine 5′-monophosphate （AMP）-activated protein kinase （AMPK）， hypoxia-inducible factor-1-BCH domain-containing protein （HIF-1-BNIP）， vascular endothelial growth factor/hypoxia-inducible factor-1α （VEGF/HIF-1α） and other signaling pathways. This review is expected to provide a theoretical basis and reference for the treatment of cardiovascular diseases in diabetes mellitus with traditional Chinese medicine.

Objective To explore the correlation of contrast-enhanced ultrasound(CEUS)parameters of adenomyoma before and after MR-guided focused ultrasound surgery(MRgFUS)with the therapeutic efficacy.Methods Uterine ultrasound and CEUS data of 26 patients with adenomyoma before and 24 h,1 and 6 months after MRgFUS,as well as MRI before and immediately after MRgFUS were retrospectively analyzed.The lesion volume shown on CEUS and MRI before MRgFUS,the non perfusion volume(NPV)of adenomyoma on MRI immediately after and CEUS 24 h after MRgFUS were compared.The ablation rate of lesions was calculated based on CEUS 24 h after MRgFUS.The focal blood flow score before,24 h after MRgFUS and the sum of the two,also the numerical rating scale(NRS)score before and 1,6 months after MRgFUS and the change rate were analyzed.The correlations of CEUS parameters with the efficacy of MRgFUS for treating adenomyoma were observed.Results No significant difference of lesion volume nor NPV on CEUS or MRI was found(both P＞0.05).The ablation rate of lesions 24 h after treatment was(58.11±24.92)%.The focal blood flow score before,24 h after MRgFUS and the sum of the two was 2.00(2.00,2.00),1.00(1.00,1.00)and 3.50(3.00,3.50),respectively,with significant difference between before and 24 h after MRgFUS(Z=-4.463,P＜0.001).NRS score was 5.00(4.00,6.00),3.00(2.00,4.00)and 2.00(1.00,3.00)before treatment,1 and 6 months after treatment,respectively,with significant differences at different time points(all P＜0.01).The change rate of NRS score 1 and 6 months after treatment was 35.42%(23.75%,50.00%)and 60.00%(50.00%,77.08%),respectively.The lesion blood flow score before and 24 h after MRgFUS and the sum of the two were all negatively correlated with ablation rate(rs=-0.552,-0.820,-0.745),while positively correlated with NRS scores 6 months after treatment(rs=0.513,0.552,0.496)but negatively correlated with the change rate of NRS scores 6 months after treatment(rs=-0.525,-0.479,-0.531).The ablation rate 24 h after treatment was negatively correlated with NRS scores(rs=-0.462)while positively correlated with the change rate of NRS scores 6 months after treatment(rs=0.500).Conclusion CEUS parameters before and after treatment were correlated with the therapeutic efficacy of MRgFUS for treating adenomyoma.