Objective: To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. Methods: Sixty C57BL/6 male mice were randomly divided into control (10 mice) and intervention groups (50 mice) using random number table method. The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg). After modeling, the intervention group was further divided into model, semaglutide (40 μg/kg), and high-, medium-, and low-dose Yishen Tongluo Formula groups (15.6, 7.8, and 3.9 g/kg, respectively) using random number table method. The high-, medium-, and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage, the semaglutide group received a subcutaneous injection of semaglutide injection, and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks. Random blood glucose levels of mice in each group were monitored, and the 24-h urinary protein content was measured using biochemical method every 4 weeks; after treatment, the serum creatinine and urea nitrogen levels were measured using biochemical method. The weight of the kidneys was measured, and the renal index was calculated. Hematoxylin and eosin, periodic acid-Schiff, periodic Schiff-methenamine, and Masson staining were used to observe the pathological changes in renal tissue. An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) levels. Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), Caspase-1, gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in renal tissue. Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4, MyD88, and NF-κB in renal tissue. Results: Compared with the control group, the random blood glucose, 24-h urinary protein, serum creatinine, urea nitrogen, and renal index of the model group increased, and the urine β2-MG, NGAL, and KIM-1 levels increased. The relative protein and mRNA expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in renal tissue increased, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas increased (P<0.05). Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group. Compared with the model group, the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment (P<0.05). The Yishen Tongluo Formula high- and medium-dose groups showed a decrease in 24-h urinary protein, creatinine, urea nitrogen, and renal index, as well as decreased β2-MG, NGAL, and KIM-1 levels. NLRP3, Caspase-1, GSDMD, IL-1 β, and IL-18 relative protein and mRNA expression levels were also reduced, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas was reduced (P<0.05). Pathological damage to renal tissue was ameliorated. Conclusion Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.

ObjectiveTo investigate the effect and mechanism of Yishen Tongluo prescription in protecting mice from oxidative stress injury in diabetic kidney disease （DKD） via the nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H quinone oxidoreductase 1 （NQO1） signaling pathway. MethodsSpecific pathogen-free （SPF） male C57BL/6 mice were assigned into a control group （n=10） and a modeling group （n=50）. The DKD model was established by intraperitoneal injection of streptozotocin. The mice in the modeling group were randomized into a model group， a semaglutide （40 μg·kg^-1） group， and high-， medium-， and low-dose （18.2， 9.1， 4.55 g·kg^-1， respectively） Yishen Tongluo prescription groups， with 10 mice in each group. The treatment lasted for 12 weeks. Blood glucose and 24-h urine protein levels were measured， and the kidney index （KI） was calculated. Serum levels of creatinine （SCr）， blood urea nitrogen （BUN）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） were assessed. The pathological changes in the renal tissue were evaluated by hematoxylin-eosin， periodic acid-Schiff， periodic acid-silver methenamine， and Masson’s trichrome staining. Enzyme-linked immunosorbent assay kits were used to measure the levels of β2-microglobulin （β2-MG）， neutrophil gelatinase-associated lipocalin （NGAL）， kidney injury molecule-1 （KIM-1）， liver fatty acid-binding protein （L-FABP）， nitric oxide synthase （NOS）， glutathione （GSH）， total antioxidant capacity （T-AOC）， and 8-hydroxy-2'-deoxyguanosine （8-OHdG）. Immunohistochemical staining was performed to examine the expression of Kelch-like ECH-associated protein 1 （Keap1） and malondialdehyde （MDA）. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of factors in the Nrf2/HO-1/NQO1 signaling pathway. ResultsCompared with the control group， the DKD model group showed rises in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with glomerular hypertrophy， renal tubular dilation， thickened basement membrane， mesangial expansion， and collagen deposition. Additionally， the model group showed elevated levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， lowered levels of GSH and T-AOC， up-regulated expression of MDA and Keap1， and down-regulated expression of Nrf2， HO-1， NQO1， and glutamate-cysteine ligase catalytic subunit （GCLC） （P<0.05）. Compared with the model group， the semaglutide group and the medium- and high-dose Yishen Tongluo prescription groups showed reductions in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with alleviated pathological injuries in the renal tissue. In addition， the three groups showed lowered levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， elevated levels of GSH and T-AOC， down-regulated expression of MDA and Keap1， and up-regulated expression of Nrf2， HO-1， NQO1， and GCLC （P<0.05）. ConclusionYishen Tongluo prescription exerts renoprotective effects in the mouse model of DKD by modulating the Nrf2/HO-1/NQO1 signaling pathway， mitigating oxidative stress， and reducing renal tubular injuries.

Objective To systematically evaluate the risk prediction models of diabetic foot infections,and to ex-plore its implications for clinical nursing care.Methods Relevant studies in the databases of CNKI,VIP,Wanfang,CBM,CINAHL,PubMed,Web of Science,Cochrane Library,Embase,ProQuest,and Ovid were searched for the time-frame of database construction to December 2023.Literature was screened independently by 2 investigators and the area under the model curve(AUC)and its 95％confidence interval(CI),calibration method and predictors were ex-tracted,and the quality of the model was evaluated using the Predictive Modeling Study Risk of Bias Assessment Tool.Meta-analysis of the predictive value of model predictors was performed using RevMan 5.4 software.Results 7 studies were included,containing 8 risk prediction models,all of which reported the AUC(0.748-0.922),and 6 models underwent model calibration.The overall fit of the included studies was good,among which l model was low fit,and all 7 studies were at high risk of bias,mainly focusing on predictors and analysis.Meta-analysis showed that the area under the curve of the combined models was 0.831(95％CI:0.780～0.883);the disease duration(OR=2.460,95％CI:1.850～3.270),vascular disease(OR=3.110,95％CI:1.070～9.000),and neuropathy(OR=3.550,95％CI:1.920～6.560)were effective predictors of diabetic foot infection.Conclusion The current diabetic foot infection risk prediction model is still in the developmental stage,and further optimization and external validation of the ex-isting model is recommended for the future.

Objective To investigate the effects and mechanism of Zhachong Shisanwei Pills on rats with cerebral ischemia.Methods Totally 75 rats were randomly divided into sham-operation group,model group,positive drug group(Ginaton,21.6 mg/kg),and Zhachong Shisanwei Pills low-,medium-,and high-dosage groups(81,162,324 mg/kg).Each treatment group was given the corresponding drug by gavage for 5 days.On the 6th day,a cerebral ischemia rat model was prepared by suture method.After 24 hours of modeling,the drugs were given in the same manner for 2 days.Neurological function scoring,horizontal beam walking scoring,and grip strength testing were performed on rats.TTC staining was used to detect the cerebral infarction rate,HE staining and Nissl staining were used to observe the morphology of brain tissue.TUNEL staining was used to detect the apoptosis rate of brain tissue cells.Differential genes in the treatment of cerebral ischemia using Zhachong Shisanwei Pills were screened by transcriptomics,and RT-qPCR,immunohistochemistry and Western blot were used to detect differential gene mRNA and protein expression.Results Compared with the sham-operation group,the model group rats showed a decrease in neurological function scores,horizontal beam walking scores,grip strength,an increase in cerebral infarction rate,neuronal nucleus condensation,vacuolar changes,widened intercellular spaces,the number of Nissl bodies reduced,and the apoptosis rate increased(P<0.01,P<0.001);compared with the model group,the Zhachong Shisanwei Pills medium-dosage group showed an increase in neurological function score,horizontal beam walking score,and grip strength in rats,a decrease in cerebral infarction rate,a lower degree of neuronal damage,an increase in the number of Nissl bodies,and a decrease in cell apoptosis rate(P<0.05,P<0.01).Transcriptome and bioinformatics analysis screened the Hippo signaling pathway related to the anti-cerebral ischemia effect of Zhachong Shisanwei Pills.The key genes of this pathway,mammalian sterile line 20 like kinase(MST1)1,Yes related protein(YAP)1,large tumor suppressor kinase(LATS)1,and TEA domain family member(TEAD)1 were detected.The results showed that the expression of MST1 mRNA and protein in brain tissue of model rats significantly increased,while the expressions of YAP1,LATS1,TEAD1 mRNA and protein significantly decreased;Zhachong Shisanwei Pills could down-regulate the expression of MST1 in brain tissue of model rats,and up-regulate the expressions of YAP1,LATS1 and TEAD1.Conclusion Zhachong Shisanwei Pills may exert anti-cerebral ischemia effects through the Hippo signaling pathway.

Objective:To construct a hypoglycemia random forest prediction model for older adults with type 2 diabetes, and assess the model′s prognostication performance through internal and external verification.Methods:From August 2022 to January 2023, 300 older adults with type 2 diabetes in Beijing Hospital were selected. The demographic characteristics, medical history, laboratory tests, and other data of the patients were collected, and the data set was randomly divided into the training set and verification set in a ratio of 7∶3. The hypoglycemia prediction model for older adults with type 2 diabetes was constructed and optimized based on the random forest algorithm. The calibration curve was used to evaluate the model′s calibration, and the ROC was used to evaluate the model′s discrimination. The clinical applicability of the model was assessed by the decision curve analysis. The risk factors for hypoglycemia in the older adults were explored by prioritizing the contributions of variables in prediction. The Bootstrap method was used for internal validation, and the validation set was used for external validation.Results:Among the 300 older adults with type 2 diabetes, 128 cases (42.67%) experienced hypoglycemia within one week. The predictive contributions of risk factors in the model were ranked as follows: the number of episodes of hypoglycemia in one month, HDL-C, heart disease, diabetes knowledge and education, combination therapy, age, duration of diabetes, staple food restriction, glycosylated hemoglobin, and gender. The internal and external calibration curves of the hypoglycemia random forest model for the older adults with type 2 diabetes fluctuated around the diagonal, indicating that the calibration degree of the predictive model is good. The AUROC of internal verification was 0.823 (95% CI 0.752-0.894), the sensitivity and specificity were 0.867 and 0.698, respectively. The external verification was 0.859 (95% CI 0.817 - 0.902), and sensitivity and specificity were 0.789 and 0.804, respectively, showing that the overall discrimination of the prediction model was good. The DCA curves were far from the all-positive line and all-negative line, which indicated that the prediction model had good clinical applicability. Conclusions:The predictive effect of this model is good, and it is suitable for predicting the risk of hypoglycemia in older adults with type 2 diabetes, and it provides a reference for early hypoglycemia screening and predictive intervention for this kind of patients.

OBJECTIVE Ulcerative colitis （UC）， as a common and refractory disease of the digestive system， has always been a hot and difficult point in medical research. Traditional Chinese medicine has the advantages of good efficacy， high safety and not easy to relapse after drug withdrawal in the treatment of UC， but the mechanism has not been fully elucidated. Metabonomics looks for potential biomarkers and metabolic pathways from the point of view of the endogenous dynamic metabolism of the whole body， which is helpful to evaluate the efficacy of drugs and explore related mechanisms. Metabolomics studies on the treatment of UC with traditional Chinese medicine have shown that traditional Chinese medicine formulas， single herbs and herbs monomers act on various related pathways such as amino acid metabolism， lipid metabolism and energy metabolism by regulating endogenous metabolites in the body， thereby inhibiting immune inflammatory reactions， improving oxidative stress， reducing intestinal sensitivity， regulating intestinal microbiota， repairing intestinal mucosal damage， and restoring normal metabolic activity in the body. However， further screening and validation of relevant metabolic markers are needed.

ObjectiveTo clone coumarate-3-hydroxylase gene （C3H） from Angelica sinensis， and analyze the correlation between its bioinformatics， expression patterns and content of ferulic acid， and to explore the functions of ASC3H. MethodReal-time polymerase chain reaction （Real-time PCR） was used to clone the full-length cDNA of ASC3H based on the transcriptome dataset of A. sinensis， and the bioinformatics analysis of the gene sequence was carried out. Real-time PCR and high performance liquid chromatography （HPLC） were used to determine relative expression of ASC3H and content of ferulic acid in different root tissues of A. sinensis （periderm， cortex and stele）. ResultThe open reading frame （ORF） of ASC3H （GenBank accession number： MN2550298） was 1 530 bp， encoding 509 amino acids， with a theoretical molecular weight of 57.86 kDa and an isoelectric point of 8.36. It was a hydrophilic protein that was located in the chloroplast with multiple phosphorylation sites and a transmembrane region， and contained a conserved domain CGYDWPKGYGPIINVW_P450 （383-399 aa） in cytochrome P450. Multiple amino acid sequence alignment analysis showed that ASC3H had high similarity with C3H from other plants， especially Ammi majus in Umbelliferae. The Real-time PCR revealed that ASC3H had different expressions in periderm， cortex and stele tissues of A. sinensis roots. It was found from HPLC that the cortex tissues had the highest content of ferulic acid， and the stele tissues had the lowest. ConclusionASC3H was successfully cloned from A. sinensis， and its sequence characteristics were understood more clearly， suggesting that ASC3H might be involved in the ferulic acid biosynthesis pathway of A. sinensis. This paper provided a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of ferulic acid in A. sinensis， while laying the foundation for the genetic improvement of A. sinensis．

OBJECTIVE: To investigate the psychological and behavior status of minor children of medical staff in Hubei province during the coronavirus disease 2019 (COVID-19) epidemic. METHODS: A cross-sectional questionnaire survey was conducted through WeChat from March 13 to 15, 2020, which included a general data questionnaire and Conners parental assessment questionnaire (PSQ). The questionnaires received from outside of Hubei province were excluded through IP address, and the questionnaires with answer time <150 s were also excluded. The influence of parental work status on the psychological behavior was analyzed in children of different age groups. RESULTS: A total of 391 valid questionnaires were collected, there were 207 males (52.9%) and 184 females (47.1%); 91 (23.3%) aged 3 to 6, 183 (46.8%) aged 6 to 10, and 117 (29.9%) aged 10 to 16. Both parents were medical staff in 87 participants(22.3%), one parent was medical staff in 139(35.5%) participants, and no parents were medical staff in 165 (42.2%) participants. In 3-<6 years group, there was no significant difference in the PSQ scores of the children in each factor level (all >0.05) between children with parents as medical staff and those without. In 6-<10 years group, children with both parents as medical staff had higher hyperactivity-impulse factor score, learning problem factor score and total score than those without parents as medical staff (all <0.05), while they had higher learning problem factor score than those with one parent as medical staff (<0.05); the anxiety score of children with one or both parents as medical staff was higher than that of those without parents as medical staff (all <0.05). In 10 to 16 years group, the behavior problems, learning problems, hyperactivity-impulse, more dynamic index and the total score in children with one parent as medical staff were lower than those with both parents as medical staff or without parents as medical staff (<0.05 or <0.01); while there were no significant differences in psychosomatic problems, anxiety factor scores between children with one parent as medical staff and other two groups (all >0.05). CONCLUSIONS s During COVID-19 epidemic period, the psychological and behavior status of minor children of Hubei medical staff with different ages shows differences with those without parents as medical staff, particularly in 6-<10 years and 10 to 16 year groups. It is necessary to pay attention to the psychological and behavioral status of children of medical staff in these age groups.
Adolescent ; Betacoronavirus ; Child ; Child, Preschool ; China ; epidemiology ; Coronavirus ; Coronavirus Infections ; Cross-Sectional Studies ; Female ; Humans ; Male ; Medical Staff ; statistics & numerical data ; Mental Health ; statistics & numerical data ; Pandemics ; Pneumonia, Viral ; Surveys and Questionnaires

Objective:To analyze the clinicopathological features, diagnosis, and treatment of anorectal malignant melanoma (ARMM), and to explore its prognostic factors and misdiagnosis. Methods:A total of 36 patients with ARMM were enrolled in this study from January 2000 to November 2016 in Nanfang Hospital, Zhujiang Hospital, and Guangdong Provincial Hospital of Traditional Chinese Medicine. Results: The clinical manifestations of ARMM were not specific. The odds of misdiagnosis were as high as 52.8% in this study. The 1-and 3-year survival rates were 75%and 35%, respectively, with median survival time of 24.51 months. Survival rate was correlated with tumor size, invasion depth, clinical stage, and lymph node metastasis (P<0.05), but was not related to patient age and gender. The median survival time of the three groups of patients (surgery alone, surgery-based combination therapy, untreated) were 39.21, 26, and 15 months. The difference was not statistically significant. No difference in survival was found between patients under-going abdominoperineal resection and wide local excision. Conclusion:ARMM has poor prognosis and is easily misdiagnosed as a ma-lignant tumor. The prognostic factors are tumor size, invasion depth, clinical stage, and lymph node metastasis. Surgical treatment can extend survival. To avoid misdiagnosis and prolong survival, early diagnosis and early treatment are recommended.

Objective To observe the effects of S100A4 siRNA on the expression of serum TNF‐α,IL‐1βand VEGF in adjuvant arthritis rats .Methods Adjuvant arthritis rat models were established and were randomly divided into model group and interfere group .On Day 11 ,rats in interfere group were injected with S100A4 siRNA fragment in articular cavity .Arthritis index (AI) chan‐ges and pathological changes of ankle joint were observed .The levels of serum TNF‐α and IL‐1β ,VEGF were detected by ELISA . Results Compared with that of model group ,the levels of serum TNF‐ α ,IL‐1β and VEGF were reduced significantly in interfere group (P< 0 .05) ;variances of AI and pathological scores in interfere group were diminished significantly (P< 0 .05) .Conclusion Inhibition of the expression of S100A4 gene can significantly reduce the expression of inflammatory factor TNF‐α ,IL‐1 β and angio‐genesis factor VEGF ,and improve the pathological injury of synovial membrane .