Objective To analyze the effect of Callicarpa nudiflora on wound healing and phos-phatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)path-way by regulating M2 macrophage polarization in rats with diabetic foot ulcer.Methods A total of 40 rats were selected,with 10 rats in control group(topical application of saline),and the remaining 30 were used to establish rat model of diabetic foot ulcer.Ultimately,27 rats were successfully mod-eled and divided into model group(topical application of saline),Callicarpa nudiflora group(topi-cal application of Callicarpa nudiflora),and Callicarpa nudiflora combined with inhibitor group(topical application of Callicarpa nudiflora combined with CD206 inhibitor,a marker protein of M2 macrophages),with 9 rats in each group.The wound healing rate,levels of basic fibroblast growth factor(bFGF)and vascular endothelial growth factor(VEGF),relative expression levels of induc-ible nitric oxide synthase(iNOS),CD16/32,CD206,PI3K,AKT and mTOR as well as mRNA ex-pression levels of PI3K,AKT and mTOR were compared among groups.Results On the 3rd,7th and 14th days after intervention,compared with the control group,the wound healing rate,VEGF,bFGF,relative expression level of CD206,relative expression of PI3K,AKT and mTOR at mRNA and protein levels were significantly decreased while the relative expression levels of iNOS and CD16/32 were significantly increased in all other groups(P＜0.05);compared with the model group,the Callicarpa nudiflora group and the Callicarpa nudiflora combined with inhibitor group showed significant increased wound healing rate,VEGF,bFGF,relative expression level of CD206,and relative expression of PI3K,AKT and mTOR at mRNA and protein levels,and significant de-creased relative expression levels of iNOS and CD16/32(P＜0.05);compared with the Callicarpa nudiflora group,the Callicarpa nudiflora combined with inhibitor group showed significant decreased wound healing rate,VEGF,bFGF,relative expression level of CD206,and relative expression of PI3K,AKT and mTOR at mRNA and protein levels,and significant increased relative expression levels of iNOS and CD16/32(P＜0.05).Conclusion Callicarpa nudiflora can promote M2 mac-rophage polarization,increase wound healing rate and levels of VEGF and bFGF in rats with diabetic foot ulcer,and activate the PI3K/AKT/mTOR signaling pathway.

Objective To explore the evaluation value of thyroid function indicators[thyroid stimulating hormone(TSH),free thyroxine(FT4),free triiodothyronine(FT3)]combined with transcranial Doppler ultrasound(TCD)and carotid ultrasound in the assessment of vascular stenosis in atherosclerotic cerebral infarction(ASCI),providing a reference for clinical targeted treatment.Methods A total of 182 patients with ASCI admitted to Handan Frist Hospital from October 2021 to September 2023 were selected and all of them underwent TCD,carotid artery ultrasound and thyroid function examination.Digital subtraction angiography was used as the gold standard to analyze the evaluation effect of TCD and carotid artery ultrasound combined with TSH,FT4 and FT3 in vascular stenosis of ASCI.Results Among the 182 patients with ASCI,DSA confirmed 64 cases(35.16％)with mild internal carotid artery stenosis,69 cases(37.91％)with moderate stenosis,29 cases(15.93％)with severe stenosis,and 20 cases(10.99％)with complete occlusion.There was no statistically significant difference in clinical data among groups with different degrees of vascular stenosis(P＞0.05).The peak systolic velocity(PSV)and end-diastolic volume(EDV)of the internal carotid artery in different groups with different degrees of vascular stenosis showed an increasing trend with the severity of vascular stenosis,while the pulse index(PI),resistance index(RI),and diameter(D)showed a decreasing trend(P＜0.05).In different groups with different degrees of vascular stenosis,TSH and FT4 levels increased with the severity of stenosis,while FT3 levels decreased(P＜0.05).TSH and FT4 were positively correlated with the degree of vascular stenosis,while FT3 was negatively correlated(P＜0.05).TSH and FT4 were positively correlated with PSV and EDV,and negatively correlated with RI,PI,and D.FT3 was positively correlated with RI,PI,and D,and negatively correlated with PSV and EDV(P＜0.05).The combined evaluation of RI,PI,D,PSV,EDV,TSH,FT4,and FT3 yielded the highest area under the curve(AUC)value of 0.941 for assessing ASCI vascular stenosis(P＜0.05).Conclusion The combination of TCD,carotid ultrasound,and thyroid function tests have high value in the assessment of ASCI vascular stenosis.The joint evaluation of RI,PI,D,PSV,EDV,TSH,FT4 and FT3 is beneficial for diagnosing different degrees of vascular stenosis,providing a reference for clinical treatment.r clinical treatment.

Objective : To construct a humanized mouse model of the interleukin-9 receptor(IL-9R) coding DNA sequence(CDS) gene and to verify the genotype and IL-9R expression in mice. Methods : The CRISPR/Cas9 genome editing technology was used to replace the exon 2-7 fragment of the il-9r gene in mouse embryonic stem cells with the corresponding human IL-9R sequence. After verifying the completion of the gene fragment replacement, tetraploid embryos were constructed and microinjected back into the oviducts of surrogate mice. Through surrogacy by female mice, homozygous humanized mice were obtained. DNA was extracted from the homozygous humanized mice IL-9R CDS gene, and their genotypes were identified by agarose gel electrophoresis after PCR amplification. Western blot was used to detect the expression of IL-9R in the spleen and thymus of homozygous humanized mice with either wild-type(WT) or IL-9R gene humanization. Results : Gel electrophoresis after PCR amplification showed that mice with only a 1 805 bp band amplified using WT primers were wild-type, while mice with 2 553 bp and 2 340 bp bands amplified using 5KI and 3KI primers, respectively, were homozygous humanized mice with IL-9R CDS gene. Western blot results indicated that the tissues of homozygous humanized mice model with IL-9R CDS gene expressed IL-9R significantly. Conclusion The humanized mouse model with IL-9R CDS gene has been successfully constructed and characterized.

Objective: To explore the breeding and genotyping of CCR2 knockout mice, and to verify the applicability of the polymerase chain reaction(PCR) method for genotype detection of CCR2 knockout mice. Methods: The introduced CCR2 pure male mice and wild-type female mice were mated and bred to produce the offspring generation, the obtained F1 generation heterozygous mice were continued to be mated. DNA was extracted by clipping the tail tissues of the mice at the age of 2 weeks, the target gene fragment was amplified by PCR, and the genotypic results were determined by agarose gel electrophoresis. The proportion of purebred progeny carrying the CCR2 knockout gene was increased by genetic crosses, the effect of CCR2 knockout in the progeny mice was verified by using Western blot against major immune cells and key organs, and flow cytometry was used to detect whether the knockout of the CCR2 gene had any effect on the function of the immune system by targeting the major immune cells. Results: CCR2 knockout mice were successfully bred and characterized, and three genotypes of F2 generation mice were obtained: CCR2+/+, CCR2+/-, and CCR2-/-. The offspring genotypes were identified by PCR, and Western blot showed extremely low CCR2 protein expression in CCR2 knockout mice. Flow analysis showed that CCR2 knockdown reduced the expression of CD4+T and Th1 cells in mouse spleen-derived T cells, but did not affect macrophage function. Conclusion Correct breeding and identification are important ways to get the pure CCR2 knockout mice, and PCR method for identifying mouse genotypes is simple, fast and reliable.

Currently,there are practical and technical difficulties in the construction of clinical ques-tions in the development of clinical practice guidelines.Clinicians or guideline developers seldom construct clin-ical questions based the actual case scenario,leading to some information loss between structured and actual clinical connotation.To overcome this challenge,we proposed a case-guided questions construction approach,and carried out case research and verification in the formulation of the guideline.We found that this method could more efficiently and scientifically assist the formulation of clinical questions,and provide reference for clinicians or guideline developers.

Objective:To analyze the clinical characteristics of patients with adverse reactions to facial skin-lightening cosmetics.Methods:A retrospective analysis was conducted on the adverse reaction reports caused by facial skin-lightening cosmetics in the cosmetic adverse reaction reporting system of Yantai city of Shandong province from July 2020 to December 2023. The general information of the patients (such as age, gender), reporting sources, clinical characteristics (types of adverse reactions, skin lesion morphology and subjective symptoms), and channels for purchasing cosmetics were summarized and analyzed.Results:A total of 450 cases of adverse reactions caused by facial skin-lightening cosmetics were identified, predominantly involving females (429 cases, 95.33%). Age distribution was most commonly found from 31 to 40 years (174 cases, 38.67%), followed by 21 to 30 years (130 cases, 28.89%), 41 to 50 years (71 cases. 15.78%), ≥51 years (46 cases, 10.22%), and ≤20 years (29 cases, 6.44%). The main sources of reporting were medical and health institutions (401 cases, 89.11%), followed by cosmetics operators (22 cases, 4.89%), patients (20 cases, 4.44%), business enterprises (3 cases, 0.67%), market supervision and administration bureaus (3 cases, 0.67%), and medical cosmetology hospital (1 case, 0.22%). Cosmetic contact dermatitis was the most common type of cosmetic adverse reaction (416 cases,91.43%), and the common skin lesions included erythema (342 cases,45.00%), papula (166 cases,21.84%), edema and so on. The common symptoms were pruritus (369 cases,49.20% ), burning sensation (158 cases,21.07%) and so on. Online sales was the main purchasing channel (333 cases, 74.01%).Conclusions:The adverse reactions caused by facial skin-lightening cosmetics are mainly found in women, and contact dermatitis is the most common type of cosmetic adverse reaction, predominantly presenting with erythema and manifesting as pruritus.

Objective:To systematically evaluate the fairness of traditional Chinese medicine (TCM) healthcare resource allocation at the prefecture-level in China using the healthcare resource agglomeration degree (HRAD) method, so as to provide empirical evidence for optimizing resource distribution.Methods:Data on TCM healthcare resources (including the number of TCM institutions, public TCM hospitals, TCM hospital beds, and TCM healthcare technicians) were collected from 333 prefecture-level regions in 2023. The HRAD method was employed to assess fairness in geographic allocation (HRAD i) and population-based allocation (HRAD i/PAD i). A multi-indicator comprehensive evaluation was conducted using the entropy weight method to determine weighting coefficients. Results:Significant disparities were observed in the geographic agglomeration of TCM resources (HRAD i) in China. Resource-rich regions (HRAD i>5) were primarily concentrated in eastern and some central-western provinces, while resource-scarce regions (HRAD i<1) were mainly distributed in western, northeastern, and parts of central and eastern provinces. Analysis of population-based fairness (HRAD i/PAD i) revealed that most prefecture-level cities nationwide had ratios<1, with only 8 provinces having more cities with ratios>1 than<1. The comprehensive evaluation showed that top-ranked cities in the east (e.g., Hangzhou, Dongying, Shenzhen), central region (e.g., Taiyuan, Zhengzhou), and west (e.g., Hainan Prefecture, Alxa League) were predominantly core cities or sparsely populated areas. Conclusions:China′s prefecture-level TCM healthcare resource allocation exhibits significant geographic and population-based inequities, with excessive concentration in provincial capitals and developed cities. Urgent strategies are needed to optimize resource allocation, enhance fairness and accessibility, including promoting the decentralization of high-quality resources, strengthening regional collaborative support, enhancing talent attraction in underdeveloped areas, and leveraging information technology to improve efficiency.

One patient with kidney stones was prescribed Paishi granules and diclofenac sodium sustained-release tablets for pain relief in the outpatient setting.That evening,the patient took 1 sachet of Paishi granules and 1 diclofenac sodium sustained-release tablet together.The patient subsequently developed a generalized rash with itching.Liver function indexes of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(T-BiL),direct bilirubin(D-BiL),and alkaline phosphatase(ALP)were all 2 to 6 times higher than the upper limit of normal.After symptomatico treatment,the patients systemic rash had subsided,and the liver function indicators have returned to normal.

Objective:To investigate the effect of glioma cell-derived apoptotic extracellular vesicles(apoEVs)on glioma tumorigen-esis and temozolomide(TMZ)resistance and its mechanism.Methods:The extracted apoEVs were characterized using nanoparticle tracking analysis and transmission electron microscopy,and Western blot,flow cytometry,CCK-8 assay,colony formation assay,and Transwell assay were used to investigate the effect of apoEVs on cell proliferation,migration,invasion,and apoptosis.Results:ApoEVs promoted TMZ resistance in glioma cells,significantly increased the half-maximal inhibitory concentration of TMZ(t=9.326,P=0.001),and inhibited cell apoptosis,and this effect could be reversed by the exosome inhibitor GW4869.ApoEVs also promoted the migration and invasion of glioma cells,increased the expression of vimentin and Twist proteins(t=8.762,P=0.002;t=7.941,P=0.004),and inhibited the expression of cleaved caspase-3(t=9.217,P=0.002).Further studies showed that apoEVs affected TMZ resistance of glioma cells by regulating the LncRNA-XIST/miR-29c/O6-methylguanine-DNA methyltransferase(MGMT)axis.Silencing of LncRNA-XIST could reduce the expression of MGMT and increase the expression of miR-29c,thereby enhancing the sensitivity to TMZ and inhibiting cell migration and invasion.Conclusion:Glioma cell-derived apoEVs promote the malignant progression and temo-zolomide resistance of glioma by delivering LncRNA-XIST to regulate the miR-29c/MGMT axis.

BACKGROUND:Wogonin is a flavonoid extracted from the root of Scutellaria baicalensis.Previous studies have shown that baicalein has protective effects against cerebral ischemia-reperfusion injury,and can also reduce blood sugar and complications in diabetic mice,but its role and mechanism in diabetic cerebral infarction remain unclear. OBJECTIVE:To explore the effect of wogonin on nerve injury in rats with diabetic cerebral infarction and its mechanism. METHODS:Sprague-Dawley rats were randomly divided into six groups:control group,model group,low-dose wogonin group,medium-dose wogonin group,high-dose wogonin group,and high-dose wogonin+Ras homolog gene family member A(RhoA)activator group.Except for the control group,the other rats were established with diabetes and cerebral ischemia models using intraperitoneal injection of streptozotocin and middle cerebral artery occlusion.Low,medium-and high-dose wogonin groups were intragastrically given 10,20,40 mg/kg wogonin,respectively;high-dose wogonin+RhoA activator group was intragastrically given 40 mg/kg wogonin and intraperitoneally injected 10 mg/kg lysophosphatidic acid;control group and model group were given the same amount of normal saline once a day for 7 consecutive days.Rats in each group were evaluated for neurological deficits and their blood glucose levels were measured after the last dose.TTC staining was applied to detect the volume of cerebral infarction.Hematoxylin-eosin staining was applied to observe pathological changes in brain tissue.ELISA kit was applied to detect tumor necrosis factor-α,interleukin-6,malondialdehyde,and superoxide dismutase levels in brain tissue.Western blot was applied to detect the protein expression of RhoA and Rho-associated protein kinase(ROCK)2 in brain tissue. RESULTS AND CONCLUSION:Compared with the control group,the neuronal structure of rats in the model group was severely damaged,with cell necrosis and degeneration,the neurological deficit score,blood glucose level,and infarct volume were significantly elevated(P<0.05),the levels of tumor necrosis factor-α,interleukin-6,and malondialdehyde,and the protein expression of RhoA and ROCK2 in brain tissue were significantly increased(P<0.05),and the superoxide dismutase level was decreased(P<0.05).Compared with the model group,the low-,medium-,and high-dose wogonin groups showed improved neuronal damage,reduced cell degeneration and necrosis,a significant reduction in neurological deficit score,blood glucose level,infarct volume,and the levels of tumor necrosis factor-α,interleukin-6,and malondialdehyde,and the protein expression of RhoA and ROCK2 in brain tissue,and an increase in the superoxide dismutase level(P<0.05).Compared with the high-dose wogonin group,the high-dose wogonin+RhoA activator group significantly weakened the improvement in the above indexes of rats with diabetic cerebral infarction(P<0.05).To conclude,wogonin can improve the blood glucose level in rats with diabetic cerebral infarction,reduce cerebral infarction and nerve injury,and its mechanism may be related to the inhibition of RhoA/ROCK signaling pathway.