BACKGROUND:Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes.The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance,but its mechanism has not been clarified.OBJECTIVE:To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.METHODS:Forty db/db mice with type 2 diabetes mellitus were randomized into a model group,a low-dose Roujishuncuiyin group,a high-dose Roujishuncuiyin group,and a positive drug group,with 10 mice in each group.The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day,respectively.In addition,10 db/dm mice were selected as the blank control group.Mice in the model and blank control groups were given the same dose of 0.9％NaCl solution by gavage.After 12 weeks of intervention,fasting blood glucose was measured in each group of mice,and oral glucose tolerance test was performed to calculate the area under the blood glucose curve.ELISA was used to detect serum insulin level and calculate the resistance index.Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy.Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B(AKT)and glycogen synthase kinase 3β(GSK-3β)proteins in skeletal muscle.RESULTS AND CONCLUSION:(1)Compared with the blank control group,fasting blood glucose,fasting insulin and insulin resistance index were significantly higher in the model group(P＜0.05),the area under the curve of the oral glucose tolerance test was significantly increased(P＜0.05),the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased(P＜0.05),and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium.(2)Compared with the model group,fasting blood glucose,fasting insulin,and insulin resistance index were significantly reduced in the low-and high-dose Roujishuncuiyin groups and the positive control group(P＜0.05),the area under the curve of the oral glucose tolerance test was reduced(P＜0.05),the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated(P＜0.05),and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated,with decreased lipid droplets in the interstitium.(3)The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group(P＜0.05),while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group(P＞0.05).To conclude,by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue,the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue,reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.

Objective:To establish and validate a prediction model based on least absolute shrinkage and selection operator（LASSO）regression and classification and regression tree（CART）algorithm for acute kidney injury（AKI）in PICU.Methods:The prospective derivation cohort consisted of 350 critically ill children admitted to the PICU of Children′s Hospital of Soochow University from September 2020 to January 2021.The external data set consisting of 866 critically ill children admitted to the PICU of Children′s Hospital of Soochow University from February 2021 to February 2022 was employed for the external validation.Clinical data was obtained from the electronic medical record system，including demographic characteristics，laboratory data and the pediatric risk of mortality Ⅲ（PRISM Ⅲ）score.The variables associated with AKI were identified using LASSO regression.Subsequently，a decision tree prediction model was built using the CART algorithm.The predictive value of decision tree prediction model was evaluated using the receiver operating characteristic（ROC）curve，calibration curve，and decision curve analysis.Results:Among the 350 children in the derivation cohort，107（30.6%）developed AKI during the PICU stay；and of 866 children in the external validation cohort，165（19.1%）developed AKI during the PICU stay.The LASSO regression screened 16 candidate variables for further analysis，and the decision tree model ultimately identified 4 variables more closely associated with AKI，including fold change in serum creatinine from baseline，urine volume，PRISM Ⅲ，and C-reactive protein.The decision tree model exhibited high accuracy with AUC of 0.92，0.88，and 0.86 in the training，internal validation，and external validation cohorts，respectively.The model demonstrated good calibration and clinical applicability based on the calibration curve and decision curve analysis.Conclusion:The decision tree model based on the 4 identified clinical indicators，including fold change in serum creatinine from baseline，urine volume，PRISM Ⅲ，and C-reactive protein，is effective for the early prediction of AKI.

Objective·To investigate the effect of Th17 cell-specific Stat3 knockout on anxiety-and depressive-like behaviors.Methods·Mice with specific Stat3 knockout in Th17 cells(Stat3fl/fl;Il17a-CreERT2),named Stat3△Il17a,and wild-type mice(Stat3fl/fl,WT)were generated through the Cre/LoxP system,and Stat3 knockout was induced by intraperitoneal injection of tamoxifen.CD4+T cells were isolated using magnetic-activated cell sorting and induced to differentiate into Th17 cells.Reverse transcription polymerase chain reaction and Western blotting were used to verify Stat3 knockout efficiency.Mice were assigned into 4 groups:WT-C group,Stat3△Il17a-C group,WT-P group,and Stat3△Il17a-P group.The periodontitis model was established in the WT-P group and the Stat3△Il17a-P group by injection of Porphyromonas gingivalis-lipopolysaccharide(P.gingivalis-LPS)into the gingival sulcus.Behavioral tests,including the open field test,elevated zero maze,and forced swimming test,were conducted to evaluate changes in anxiety-and depressive-like behaviors.Micro-computed tomography was used to observe destruction of alveolar bone.Neuronal injury was observed by H-E staining,and the level of brain-derived neurotrophic factor(BDNF)was detected by enzyme-linked immunosorbent assay.Results·mRNA expression and protein levels of Stat3 in Stat3△Il17a mice were inhibited compared to WT mice(P<0.05).Experimental periodontitis model was successfully established in the WT-P group and the Stat3△Il17a-P group.The degree of alveolar bone destruction was reduced in the Stat3△Il17a-P group compared to the WT-P group.In the WT-P group,decreased residence time in the central area and in the open area was observed in the open field test and elevated zero maze respectively,and increased immortal time was recorded in the forced swimming test(P<0.05).Moreover,neuronal injury was detected and significantly decreased expression levels of BDNF in the brain were measured in the WT-P group compared with the WT-C group(P<0.05).The degree of abnormal behaviors and neuronal injury was reduced in the Stat3△Il17a-P group compared to the WT-P group(P<0.05).Moreover,the level of BDNF in the Stat3△Il17a-P was higher than that in the WT-P group(P<0.05).Conclusion·Periodontitis may contribute to anxiety-and depressive-like behaviors and neuronal damage in mice,while Th17-specific conditional knockout of Stat3 could significantly alleviate pathological behaviors and neuronal damage.Stat3-mediated-Th17 cell immune responses may play a crucial role in the correlation between periodontitis and anxiety and depression.

As key cells of innate immunity, macrophages play a crucial role in maintaining immune homeostasis through their M1/M2 polarization states. Various metabolic diseases such as type 2 diabetes mellitus and obesity are closely associated with inflammation, and macrophage polarization imbalance is commonly observed during their progression. Regulating M1/M2 macrophage polarization is highly likely to become a potential therapeutic direction for metabolic inflammation-related diseases. Through literature research, this article summarizes the origin, activation, and polarization mechanisms of macrophages, and reviews the latest research progress on the imbalance and regulation of macrophage polarization in metabolic inflammation-related diseases. It aims to provide a theoretical basis for in-depth exploration of the molecular mechanisms and precise targets of macrophage polarization, as well as optimized strategies for the prevention and treatment of metabolic inflammation-related diseases.

ObjectiveTo systematically evaluate the efficacy and safety of Qi-reinforcing and blood-activating/stasis-expelling Chinese patent medicines in the treatment of coronary microvascular disease （CMD）. MethodsPubMed， Cochrane Library， CNKI， Wanfang Data， and VIP were searched for the randomized controlled trials （RCTs） on the treatment of CMD with Chinese patent medicines for reinforcing Qi and activating blood/expelling stasis with the time interval from inception to December 31， 2023. The primary outcome indicators included the index of microcirculatory resistance （IMR）， coronary flow reserve （CFR）， and corrected TIMI flow frame count （cTFC）. The secondary outcome indicators included symptomatic efficacy， left ventricular ejection fraction （LVEF）， hypersensitive C-reactive protein （hs-CRP）， nitric oxide （NO）， and adverse events. Cochrane risk-of-bias assessment tool 2.0 （RoB 2.0） and Stata 17.0 were used for literature quality evaluation and meta-analysis of the included RCTs. The Grading of Recommendations， Assessment， Development and Evaluation （GRADE） was used to evaluate the quality of evidence. ResultsA total of 36 RCTs were included in this study， involving 3 029 patients. Compared with conventional Western medicine alone， the combined use of Chinese patent medicines for reinforcing Qi and activating blood/expelling stasis and Western medicine reduced the IMR ［mean difference （MD）=-5.93， 95% confidence interval （95%CI） ［-8.73，-3.14］， n=382， P<0.01］， cTFC （MD=-9.35， 95%CI ［-13.94，-4.76］， n=618， P<0.01）， and hs-CRP ［standard mean difference （SMD）=-1.50， 95%CI ［-1.90，-1.11］， n=1 483， P<0.01］， improved the CFR （SMD=1.14， 95%CI ［0.08，2.19］， n=304， P=0.03）， symptomatic efficacy ［relative risk （RR）=1.36， 95%CI ［1.21，1.53］， n=756， P<0.01］， LVEF （MD=4.39， 95%CI ［2.31，6.47］， n=533， P<0.01）， and NO （SMD=3.16， 95%CI ［2.07，4.25］， n=946， P<0.01） of CMD patients. In terms of safety， the combined therapy reduced the occurrence of adverse events in CMD patients （RR=0.49， 95%CI ［0.29，0.82］， n=591， P=0.01）. GRADE showed moderate quality evidence for adverse events， low quality evidence for cTFC， symptomatic efficacy， LVEF， and NO， and very low quality evidence for IMR， CFR， and hs-CRP. ConclusionBased on microcirculatory function indicators， the combined use of Qi-reinforcing and blood-activating/stasis-expelling Chinese patent medicines and Western medicine may further improve the coronary microvascular function in CMD patients with good safety. The above conclusions remain to be verified with high-quality clinical trials.

Objective:To explore the trajectory and influencing factors of oral frailty in first-onset elderly patients with ischemic stroke.Methods:A total of 240 inpatients from the neurology departments of Wutaishan Hospital and Northern Jiangsu People's Hospital between October 2022 and February 2024 were selected by convenience sampling. Oral frailty scores were assessed dynamically at four time points: 1 day before discharge (T 1) , 1 month after discharge (T 2) , 3 months after discharge (T 3) , and 6 months after discharge (T 4) . Latent growth modeling was used to identify potential trajectory categories and analyze their influencing factors. Results:A total of 240 questionnaires were distributed, with 220 valid responses, yielding a response rate of 91.67%. A total of three trajectory types of oral frailty were identified: non-oral frailty group ( n=92) , low-decline group ( n=66) , and high-decline group ( n=62) . Age, marital status, smoking history, dysphagia, National Institute of Health Stroke Scale score, oral health, nutritional status, oral health-related self-efficacy, monthly per capita household income, polypharmacy, denture use, and cognitive function were influencing factors of oral frailty in first-onset elderly patients with ischemic stroke ( P<0.05) . Conclusions:From 1 day before discharge to 6 months after discharge, there are 3 distinct oral frailty trajectories in first-onset elderly patients with ischemic stroke. Medical staff should implement individualized interventions based on the identified influencing factors.

Objective To evaluate the efficacy and safety of Jiawei Dihuang Decoction in the treatment of vascular dementia(VD);To explore its mechanism and the VD susceptibility genesby using whole exome sequencing.Methods A total of 75 patients with VD who were hospitalized or received outpatient treatment at The Second Affiliated Hospital of Shaanxi University of Chinese Medicine were included.They were divided into the control group(37 cases,treated with conventional Western medicine)and the experimental group(38 cases,treated with conventional Western medicine+Jiawei Dihuang Decoction)using random number table method.The treatment course was 3 months.The general data,TCM syndrome scores,MMSE scores,ADL scores,Blessed scores and adverse reactions of the two groups were compared.Peripheral blood samples from 36 patients with kidney-yin deficiency type VD were selected for whole exome sequencing.Susceptible genes were screened,and the targets of Jiawei Dihuang Decoction were analyzed by network pharmacology.A"drug-gene"network was constructed,and key pathways were enriched.Results There was no statistical significance in the baseline data between the two groups(P>0.05),and they were comparable.Compared with before treatment,the MMSE scores of patients in both groups significantly increased after treatment,while TCM syndrome scores and ADL scores significantly decreased(P<0.05).Compared with the control group,the TCM syndrome scores,MMSE scores,ADL scores and clinical efficacy of the experimental group were significantly better than those of the control group(P<0.05).Moreover,the Blessed score showed that the experimental group had more advantages in improving the patients'living ability and daily habits(P<0.05).No adverse reactions were observed in both groups during the treatment period.A total of 1 250 744 single nucleotide polymorphism(SNP)loci and 37 314 insertion and deletion(InDel)loci were detected by whole exome sequencing.After screening,3 041 VD susceptibility genes were obtained.It was found that they were involved in biological processes such as the response to nutrient levels,positive regulation of the MAPK cascade,vascular processes in the circulatory system,the response to nutrients,etc.And enriched in PI3K-Akt,cholinergic/glutamatergic synapses,lipid metabolism and atherosclerosis pathways.The potential targets of 854 of Jiawei Dihuang Decoction were intersected with the susceptibility genes to obtain 353 common targets.The top 10 key genes were analyzed and found to be involved in positive regulation of cytosine-serine phosphorylation,miRNA-mediated gene silencing regulation,and the response of cells to decreased oxygen levels,etc.They were enriched in PI3K-Akt,lipid and atherosclerosis signaling pathways.Conclusion Jiawei Dihuang Decoction can alleviate the symptoms of patients with VD,protect cognitive function,enhance their ability of daily living,and has good safety profile.Its mechanism may involve regulating susceptibility genes through PI3K-Akt signaling pathway and lipid atherosclerosis signaling pathway,and improving lipid metabolism,inflammatory response and oxidative stress.

BACKGROUND:Skeletal muscle insulin resistance is the key pathological link of type 2 diabetes.The traditional Chinese medicine compound Roujishuncuiyin can effectively improve skeletal muscle insulin resistance,but its mechanism has not been clarified.OBJECTIVE:To explore the mechanism of Roujishuncuiyin on skeletal muscle insulin resistance in type 2 diabetes mice.METHODS:Forty db/db mice with type 2 diabetes mellitus were randomized into a model group,a low-dose Roujishuncuiyin group,a high-dose Roujishuncuiyin group,and a positive drug group,with 10 mice in each group.The latter three administration groups were given 157.5 mg/g and 630 mg/g Roujishuncuiyin and 200 mg/g metformin hydrochloride aqueous solution by gavage once a day,respectively.In addition,10 db/dm mice were selected as the blank control group.Mice in the model and blank control groups were given the same dose of 0.9％NaCl solution by gavage.After 12 weeks of intervention,fasting blood glucose was measured in each group of mice,and oral glucose tolerance test was performed to calculate the area under the blood glucose curve.ELISA was used to detect serum insulin level and calculate the resistance index.Mitochondrial structure of skeletal muscle tissue was observed under transmission electron microscopy.Western blot was used to detect the expression levels and phosphorylation levels of protein kinase B(AKT)and glycogen synthase kinase 3β(GSK-3β)proteins in skeletal muscle.RESULTS AND CONCLUSION:(1)Compared with the blank control group,fasting blood glucose,fasting insulin and insulin resistance index were significantly higher in the model group(P＜0.05),the area under the curve of the oral glucose tolerance test was significantly increased(P＜0.05),the expression of p-AKT and p-GSK3β proteins in tibialis anterior muscle was significantly decreased(P＜0.05),and there was a large amount of mitochondrial damage in tibialis anterior muscle and a large number of lipid droplets in the interstitium.(2)Compared with the model group,fasting blood glucose,fasting insulin,and insulin resistance index were significantly reduced in the low-and high-dose Roujishuncuiyin groups and the positive control group(P＜0.05),the area under the curve of the oral glucose tolerance test was reduced(P＜0.05),the expression of p-AKT and p-GSK3β proteins in the tibialis anterior muscle was significantly elevated(P＜0.05),and mitochondrial damage in the tibialis anterior muscle was significantly ameliorated,with decreased lipid droplets in the interstitium.(3)The above indexes were better in the high-dose Roujishuncuiyin group than the low-dose Roujishuncuiyin group(P＜0.05),while there was no significant difference between the high-dose Roujishuncuiyin group and positive control group(P＞0.05).To conclude,by upregulating the protein levels of p-AKT and p-GSK3β in skeletal muscle tissue,the traditional Chinese medicine compound Roujishuncuiyin can improve structural disorders and mitochondrial morphology in skeletal muscle tissue,reduce insulin resistance in the skeletal muscle and regulate glucose homeostasis in the body.

Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Ultrasound-guided peripheral nerve block is widely used in perioperative pain management of hip fracture patients,which not only provides better analgesia,but also reduces the use of opioids,while tradi-tional techniques such as femoral nerve block and iliofascial space block often result in motor block,which is not conducive to the early rehabilitation of patients.The newly proposed pericapsular nerve block and iliopsoas plane block of the hip can preserve motor function while providing satisfactory analgesia,further in-depth re-search has found that it has great significance in the diagnosis and treatment process of patients with hip frac-tures.This article reviews the innervation of the hip joint,the methods of operation and clinical application of ultrasound-guided pericapsular nerve block and iliopsoas plane block of the hip joint,in order to provide refer-ence for clinical practice.