Objective To establish an epirubicin (EPI)-resistant murine triple-negative breast cancer (TNBC) (4T1/EPI) cell line and evaluate its biological characteristics and drug resistance. Methods The EPI-resistant cell line 4T1/EPI was developed through intermittent induction with gradually increasing EPI concentrations in vitro. Morphological changes were observed under an inverted microscope. Drug resistance index (MTT assay), cell doubling time (CCK-8 assay), and migration ability (wound healing assay) were evaluated. Western blot was used to detect the expression of drug resistance-related proteins. Transcriptome sequencing and KEGG pathway enrichment analysis were performed to identify the pathways and targets involved in EPI resistance, followed by experimental validation. Results The 4T1 cells eventually grew normally in a medium containing 100 ng/mL EPI, confirming the establishment of the 4T1/EPI resistant cell line. After stable resistance was acquired, morphological alterations were observed. Compared with their parental 4T1 cells, 4T1/EPI cells showed significantly prolonged doubling time (P<0.01) and enhanced migration ability (P<0.05). Expression levels of drug resistance-related proteins MDR1, MRP1 (P<0.01), and ABCG2 (P<0.05) were elevated in 4T1/EPI cells. In vivo models also demonstrated significant EPI resistance in 4T1/EPI tumors in terms of tumor weight and volume. Transcriptome sequencing highlighted the involvement of the PI3K/Akt signaling pathway and ABC transporter pathway. Validation experiments showed the upregulation of Erbb3, Egfr, PI3K, and Akt (P<0.05) and significant downregulation of Fgfr1 (P<0.01) in 4T1/EPI cells. Conclusion The EPI-resistant TNBC cell line 4T1/EPI was successfully established, exhibiting significant resistance in vitro and in vivo. The mechanism may involve the EPI-induced upregulation of Egfr and Erbb3, activating the PI3K/Akt pathway and subsequently enhancing ABC transporter expression.

OBJECTIVE To explore the effectiveness and possible mechanism of Shangke Lengtong Patch in treating knee osteo-arthritis with cold-dampness arthralgia obstruction.METHODS A total of 70 patients who met the inclusion criteria of knee osteoar-thritis with cold-dampness arthralgia obstruction in the Orthopedics Department of Affiliated Hospital of Nanjing University of Chinese Medicine from November to December 2024 were randomly divided into an experimental group and a control group,with 35 cases in each group.During the treatment,1 case dropped out of the experimental group,3 cases dropped out of the control group,and 1 case was discontinued.The experimental group was treated with Shangke Lengtong Patch,and the control group was treated with Compound Nanxing Zhitong Ointment.The WOMAC scores and TCM syndrome scores of the two groups before and after treatment were compared to comprehensively evaluate the clinical efficacy.The changes in the expression levels of CGRP,substance P,HMGB1,IL-1β,CX-CL12,and CXCR4 in the serum of the two groups were detected by ELISA.RESULTS After 3,7,and 14 d of treatment,the WOMAC scores and TCM syndrome scores of the two groups were significantly reduced(P<0.05,P<0.01),and the score of aggrava-ted cold in the experimental group was better than that in the control group at 7 d of treatment(P<0.05);after 14 d of treatment,the expression levels of CGRP,substance P,HMGB1,IL-1β,CXCL12,and CXCR4 in the serum of the two groups were significantly re-duced(P<0.05),and there was no statistical difference between the two groups.CONCLUSION Shangke Lengtong Patch can sig-nificantly relieve the pain symptoms of knee osteoarthritis patients with cold-dampness arthralgia obstruction and improve the joint func-tion of patients.It may improve synovial inflammation by inhibiting the HMGB1/CXCL12/CXCR4 pathway,thereby exerting a thera-peutic effect.

Recent studies have shown that the physiological homeostasis of oral mucosal cells is regulated by the circadian clock.Dis-ruption or dysfunction of the circadian clock is closely associated with the development of oral squamous cell carcinoma(OSCC).Research based on the circadian clock offers a novel perspective on the pathogenesis and therapeutic strategies for OSCC.However,there is current-ly limited research on this topic,and people generally have insufficient understanding and recognition of the circadian clock.Given the complexity and challenges of circadian clock which is the fourth dimension of medical research,we organize relevant experts based on summarizing the current research results of circadian clock in the pathogenesis and precision diagnosis and treatment of OSCC,combining the scientific principles of the circadian clock's role and their long-term research experience,then summarizes and recommends the con-sensus opinions for the research of circadian clock in the pathogenesis mechanism and precision diagnosis and treatment of human OSCC,with the hope of providing guidance for the basic research and clinical application of circadian clock or circadian rhythm in the pathogene-sis mechanism and precision diagnosis and treatment of oral and maxillofacial squamous cell carcinoma.

Objective To evaluate the clinical efficacy of hip-knee-ankle active and passive exercise therapy in patients with early-to mid-stage knee osteoarthritis(KOA).Methods A total of 180 patients with early to mid-stage knee osteoarthritis(KOA)were recruited from the First Affiliated Hospital of Hebei University of Tradi-tional Chinese Medicine between March 2023 and March 2024.Patients were randomly assigned to one of four groups:active movement group,passive movement group,combined movement group,and control group,with 45 patients in each group.The active movement group received hip-knee-ankle active movement therapy daily until the end of follow-up.The passive movement group underwent hip-knee-ankle passive movement therapy three times per week for two weeks.The combined movement group received both active and passive therapies.The control group was administered oral celecoxib capsules(200 mg once daily for two weeks).Joint function was assessed in all four groups before treatment,at two weeks post-treatment,and at 14 weeks post-treatment.The primary outcome measure was the WOMAC joint function score,while secondary outcomes included the WOMAC pain score,stiffness score,and quality of life score(SF-12).Results A total of 160 patients completed the trial,with 39 in the active group,42 in the passive group,40 in the combined group,and 39 in the control group.There were no significant differences in baseline characteristics among the groups(P>0.05).Compared to baseline,the WOMAC scores for function,pain,and stiffness in the passive,combined,and control groups decreased significantly at both 2 and 14 weeks post-treatment(P<0.05),while the SF-12 scores increased significantly(P<0.05).Between 2 and 14 weeks post-treat-ment,the active and combined groups showed further significant decreases in WOMAC function,pain,and stiffness scores(P<0.05)and increases in SF-12 scores(P<0.05).At 2 weeks post-treatment,compared to the control group,the passive and combined groups exhibited significantly lower WOMAC function scores(P<0.05),with no significant difference between the passive and combined groups(P>0.05).By 14 weeks post-treatment,the active and combined groups demonstrated significantly lower WOMAC function scores(P<0.05),with the combined group showing a significantly lower score than the active group(P<0.05).Conclusion The four therapeutic approaches demonstrate a certain degree of efficacy in improving joint function for patients with early and mid-stage KOA.The passive therapy group exhibits superior short-term outcomes,while the active therapy group shows better long-term benefits.The combined therapy group presents notable advantages in both short-term and long-term effi-cacy,although its short-term effectiveness does not surpass that of the passive therapy group.It is recommended for patients with early and mid-stage KOA who have underlying gastrointestinal and cardiovascular conditions.

Objective To compare the success rate and stability of rat models of comorbid chronic pain and depression induced by different doses of complete Freund's adjuvant(CFA).Methods Sixty SD rats were divided randomly into a control group,low-dose CFA group(CFA-L),and high-dose CFA group(CFA-H)(n=20 rats per group).Rats in the CFA-L and CFA-H groups were injected with 50 and 100 μL CFA,respectively,and rats in the control group were injected with 0.9％sodium chloride solution.The general state,body weight,mechanical withdrawal threshold(MWT),and thermal withdrawal latency(TWL)were observed at 0,7,14,21,and 28 days after modeling.Depressive behavior was evaluated using the open field test(OFT),forced swim test(FST),and tail suspension test(TST).Glutamate(Glu)and γ-aminobutyric acid(GABA)levels in the anterior cingulate cortex were detected by enzyme-linked immunosorbent assay.Brain-derived neurotrophic factor(BDNF)expression in the anterior cingulate cortex was detected by immunohistochemistry,and pathological changes in the anterior cingulate cortex were observed by HE staining.Results(1)Regarding the general condition of the rats,the left ankle joint and toes were obviously red and swollen in the CFA-L and CFA-H groups on the 7th day after modeling,and the swelling was more severe in the CFA-H group.The redness and swelling of the left hind foot and ankle joint and toes gradually recovered in the CFA-L group on days 14,21,and 28 after modeling,but were still obvious in the CFA-H group,and the water and food intake decreased.(2)The body mass was significantly lower in rats in the CFA-H group compared with those in the blank and CFA-L groups on days 14,21,and 28 after modeling(P＜0.05,P＜0.05).(3)Regarding pain-related behavior,the MWT and TWL were significantly decreased in the CFA-L and CFA-H groups on the 7th and 14th days after modeling,compared with the control group(P＜0.05,P＜0.05).On day 21 after modeling,MWT was significantly lower in the CFA-H group than in the blank and CFA-L groups(P＜0.05,P＜0.05),and TWL was significantly lower in the CFA-L and CFA-H groups than in the blank group(P＜0.05,P＜0.05).On day 28 after modeling,MWT and TWL were significantly lower in the CFA-H group than in the blank and CFA-L groups(P＜0.05,P＜0.05).(4)In terms of depression-related behaviors,the total OFT movement distance was significantly lower in the CFA-H group than in the blank and CFA-L groups on day 7 after modeling(P＜0.05,P＜0.05).The total OFT distance and central dwell time were significantly lower in the CFA-H group than in the blank and CFA-L groups on days 14,21,and 28 after modeling(P＜0.05,P＜0.05),and the result in the FST and TST were significantly higher than in the blank and CFA-L groups(P＜0.05,P＜0.05).(5)Glu,GABA,and BDNF expression levels were significantly higher in the CFA-H group than in the blank and CFA-L groups(P＜0.05,P＜0.05),while GABA,Glu/GABA,and BDNF levels were significantly lower in the CFA-H group than in the blank and CFA-L groups(P＜0.05,P＜0.05,P＜0.05).(6)The CFA-L group showed less damage in the anterior cingulate cortex,more pyramidal cells,more arranged cells,clear nucleoli,and a small number of cells with karyokynesis and deep staining.Compared with the CFA-L group,rats in the CFA-H group showed a disordered cell arrangement in the injured area of the anterior cingulate cortex,a large number of pyknotic and hyperchromatic neurons,significantly fewer or absent pyramidal cells,and vacuoles,red blood cells,and neurofibrillary tangles in the interstitial space.Conclusions Injection of CFA 100 μL can be used to establish a rat model of chronic inflammatory pain and depression,showing hyperalgesia,depression-like behavioral changes,changes in levels of Glu,GABA,and BDNF in the anterior cingulate cortex,and pathological changes in the anterior cingulate cortex,consistent with the pathophysiological characteristics of chronic pain and depression.

Objective To investigate the changes and clinical significance of serum C1q tumor necrosis factor-related protein 12(CTRP12),adiponectin(APN),and resistin(RETN)in patients with type 2 diabetes mellitus(T2DM)combined with different grades of hypertension.Methods A total of 156 patients with T2DM were selected and divided into the following groups:isolated T2DM group(T2DM,n=38),T2DM combined with hypertension grade 1(DH1,n=39)group,T2DM combined with hypertension grade 2(DH2,n=39)group,and T2DM combined with hypertension grade 3(DH3,n=40)group.Additionally,healthy individuals who underwent physical examinations at our hospital were selected as the normal control(NC,n=40)group.Serum levels of CTRP12,APN,and RETN were measured using ELISA.Pearson correlation analysis was used to evaluate the relationship between serum CTRP12,APN,RETN,and other indicators.Multiple linear regression analysis was performed to determine the influencing factors for the three indicators.Results In NC,T2DM,DH1,DH2 and DH3 groups,DBP increased in turn(P<0.05),while CTRP12 decreased in turn(P<0.05).SBP in T2DM,DH1,DH2 and DH3 groups increased in turn(P<0.05).In DH1,DH2 and DH3 groups,RENT increased in turn(P<0.05),while APN decreased in turn(P<0.05).Pearson correlation analysis showed that serum CTRP12 was negatively correlated with WHR,BMI,HOMA-IR,FPG,HbA1c,FIns,TC,LDL-C,SBP,and DBP(P<0.05).Serum APN was negatively correlated with BMI,FPG,HbA1c,FIns,TC,SBP,and DBP(P<0.05),and positively correlated with HDL-C(P<0.05).Serum RETN was positively correlated with WHR,HOMA-IR,FPG,HbA1c,FIns,SBP,and DBP(P<0.05).Multiple linear regression analysis indicated that SBP was an influencing factor for CTRP12 and RETN,while SBP and HbA1c were influencing factors for RETN.Conclusions In patients with T2DM combined with hypertension,serum CTRP12 and APN levels decrease,while RETN levels increase,with significant changes observed as HP grades increase,suggesting that these three factors may collectively influence the occurrence and development of T2DM combined with HP.

Objective:To screen high risk children for lysosomal storage diseases (LSD) in southern China and analyze the spectrum characteristics of LSD in this region.Methods:A cross-sectional study was conducted. A total of 7 435 children at high risk of LSD were screened at Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from January 2009 to December 2024. The activities of 22 lysosomal enzymes from peripheral blood leukocytes or plasma were measured by fluorescence or colorimetric assays with synthetic substrates to screen for 24 LSD subtypes.Results:Among the 7 435 high risk children, 759 children were diagnosed with LSD (10.2%). The diagnosed cases included 506 males and 253 females, with an age at diagnosis of 3.0 (2.5, 5.5) years. The common disease types were mucopolysaccharidosis (MPS) (390 cases (51.4%)), sphingolipidoses (269 cases (35.4%)), glycogen storage disease (62 cases (8.2%)), and mucolipidosis types Ⅱ and Ⅲ (29 cases (3.8%)). Among the positive cases, 21 disease subtypes were identified. The 5 frequent subtypes, in descending order, were MPS type Ⅱ (197 cases (26.0%)), Gaucher disease (111 cases (14.6%)), MPS type ⅣA (87 cases (11.5%)), glycogen storage disease type Ⅱ (62 cases (8.2%)), and metachromatic leukodystrophy (MLD) (49 cases (6.5%)). The rarest subtypes were mannosidosis, multiple sulfatase deficiency and Wolman disease, each with 1 case (0.1%).Conclusions:Enzyme activity screening is essential for diagnosing high risk children with LSD. In Southern China, the most common LSD subtypes are MPS Ⅱ, Gaucher disease, MPS ⅣA, glycogen storage disease type Ⅱ, and MLD, while mannosidosis, multiple sulfatase deficiency and Wolman disease are the rarest.

To effectively carry out community health education and prevention and control of chronic obstructive pulmonary disease(COPD),we conducted a questionnaire survey on the awareness of COPD knowledge among residents aged 40 and above in Jiading District of Shanghai by using two-stage sampling method.We included 1 783 permanent residents in 4 districts and collected 1 666 valid questionnaires.The awareness rates of COPD name,pulmonary function test and related knowledge were 15.9%,11.9%and 17.2%,respectively.For the main symptoms of COPD,the awareness rate of sputum was the lowest(65.2%),of shortness of breath and dyspnea was the highest(78.4%).For the risk factors,the awareness rate of severe respiratory tract infection in childhood was the lowest(67.3%),of smoking was the highest(86.8%).Multiple response analysis showed that mobile phones and computers accounted for the highest proportion of acquiring knowledge of COPD(66.7%).Multivariate logistic regression analysis showed that low educational level,low monthly income,engaged in production and manufacturing,never or less exercise,no use of indoor air improvement measures,no wearing of masks in public places and no vaccination of influenza vaccine per year were associated with lower awareness rate of COPD.The community should improve the way and ability of health science popularization,and carry out health education for key groups.

Obstructive sleep apnea (OSA) is an increasingly widespread sleep-breathing disordered disease, and is an independent risk factor for many high-risk chronic diseases such as hypertension, coronary heart disease, stroke, arrhythmias and diabetes, which is potentially fatal. The key to the prevention and treatment of OSA is early diagnosis and treatment, so the assessment and diagnostic technologies of OSA have become a research hotspot. This paper reviews the research progresses of severity assessment parameters and diagnostic technologies of OSA, and discusses their future development trends. In terms of severity assessment parameters of OSA, apnea hypopnea index (AHI), as the gold standard, together with the percentage of duration of apnea hypopnea (AH%), lowest oxygen saturation (LSpO₂), heart rate variability (HRV), oxygen desaturation index (ODI) and the emerging biomarkers, constitute a multi-dimensional evaluation system. Specifically, the AHI, which measures the frequency of sleep respiratory events per hour, does not fully reflect the patients’ overall sleep quality or the extent of their daytime functional impairments. To address this limitation, the AH%, which measures the proportion of the entire sleep cycle affected by apneas and hypopneas, deepens our understanding of the impact on sleep quality. The LSpO₂ plays a critical role in highlighting the potential severe hypoxic episodes during sleep, while the HRV offers a different perspective by analyzing the fluctuations in heart rate thereby revealing the activity of the autonomic nervous system. The ODI provides a direct and objective measure of patients’ nocturnal oxygenation stability by calculating the number of desaturation events per hour, and the biomarkers offers novel insights into the diagnosis and management of OSA, and fosters the development of more precise and tailored OSA therapeutic strategies. In terms of diagnostic techniques of OSA, the standardized questionnaire and Epworth sleepiness scale (ESS) is a simple and effective method for preliminary screening of OSA, and the polysomnography (PSG) which is based on recording multiple physiological signals stands for gold standard, but it has limitations of complex operations, high costs and inconvenience. As a convenient alternative, the home sleep apnea testing (HSAT) allows patients to monitor their sleep with simplified equipment in the comfort of their own homes, and the cardiopulmonary coupling (CPC) offers a minimal version that simply analyzes the electrocardiogram (ECG) signals. As an emerging diagnostic technology of OSA, machine learning (ML) and artificial intelligence (AI) adeptly pinpoint respiratory incidents and expose delicate physiological changes, thus casting new light on the diagnostic approach to OSA. In addition, imaging examination utilizes detailed visual representations of the airway’s structure and assists in recognizing structural abnormalities that may result in obstructed airways, while sound monitoring technology records and analyzes snoring and breathing sounds to detect the condition subtly, and thus further expands our medical diagnostic toolkit. As for the future development directions, it can be predicted that interdisciplinary integrated researches, the construction of personalized diagnosis and treatment models, and the popularization of high-tech in clinical applications will become the development trends in the field of OSA evaluation and diagnosis.

Allergic rhinitis(AR)affects 10%-40%of the population in the world.Because CLOCK genes regulate the circadian rhythms of AR,the symptoms in parts of the patients are aggravated in the morning and evening and relieved at moon.Although lots of studies showed that traditional Chinese medicine is effective for AR,further explores are still needed to analyze the circadian rhythm of AR based on Chinese Chronological Medicine for controlling the circadian outbreak point of symptoms of AR in daily cycle.We supposed that Weiyang might play an important role for controlling the circadian rhythm of AR with closed similarities with the tidal variation of the expressions of CLOCK genes.Thus based on Weiyang theory,we tried to clarify the mechanism of circadian rhythm of AR and explore the relationship between Weiyang and the CLOCK genes so as to further expand the treatment methods of adjusting the circadian cycle of AR with TCM and improve the corresponding therapeutic efficacy.