Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

Traditional Chinese medicine (TCM) is a well-accepted therapy for atopic dermatitis (AD). However, there are currently no evidence-based guidelines integrating TCM and Western medicine for the treatment of AD, limiting the clinical application of such combined approaches. Therefore, the China Association of Chinese Medicine initiated the development of the current guideline, focusing on key issues related to the use of TCM in the treatment of AD. This guideline was developed in accordance with the principles of the guideline formulation manual published by the World Health Organization. A comprehensive review of the literature on the combined use of TCM and Western medicine to treat AD was conducted. The findings were extensively discussed by experts in dermatology and pharmacy with expertise in both TCM and Western medicine. This guideline comprises 23 recommendations across seven major areas, including TCM syndrome differentiation and classification of AD, principles and application scenarios of TCM combined with Western medicine for treating AD, outcome indicators for evaluating clinical efficacy of AD treatment, integration of TCM pattern classification and Western medicine across disease stages, daily management of AD, the use of internal TCM therapies and proprietary Chinese medicines, and TCM external treatments. Please cite this article as: Du XR, Wu MY, Tao MC, Lin Y, Gu CY, Wu MF, Cao Y, Chen DC, Li W, Wang HW, Wang Y, Wang Y, Lu HZ, Liu X, Su XF, Li FL. Clinical practice guidelines for the diagnosis and treatment of atopic dermatitis with integrative traditional Chinese and Western medicine. J Integr Med. 2025; 23(6):641-653.
Dermatitis, Atopic/drug therapy* ; Humans ; Medicine, Chinese Traditional/methods* ; Integrative Medicine ; Drugs, Chinese Herbal/therapeutic use* ; Practice Guidelines as Topic

The structure of intestinal tissue is complex. In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases. Recently, organoids have been successfully constructed and they have come to play an important role in biomedical research. Organoids are miniaturized three-dimensional (3D) organs, derived from stem cells, which mimic the structure, cell types, and physiological functions of an organ, making them robust models for biomedical research. Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine, thereby providing a valuable platform for intestinal development and disease research. In this article, we review the latest progress in the construction and application of intestinal organoids.
Organoids/cytology* ; Intestines/physiology* ; Humans ; Animals ; Pluripotent Stem Cells

OBJECTIVE: To investigate the relationship between physical activity and genetic risk and their combined effects on the risk of developing chronic obstructive pulmonary disease. METHODS: This prospective cohort study included 318,085 biobank participants from the UK. Physical activity was assessed using the short form of the International Physical Activity Questionnaire. The participants were stratified into low-, intermediate-, and high-genetic-risk groups based on their polygenic risk scores. Multivariate Cox regression models and multiplicative interaction analyses were used. RESULTS: During a median follow-up period of 13 years, 9,209 participants were diagnosed with chronic obstructive pulmonary disease. For low genetic risk, compared to low physical activity, the hazard ratios ( HRs) for moderate and high physical activity were 0.853 (95% confidence interval [ CI]: 0.748-0.972) and 0.831 (95% CI: 0.727-0.950), respectively. For intermediate genetic risk, the HRs were 0.829 (95% CI: 0.758-0.905) and 0.835 (95% CI: 0.764-0.914), respectively. For participants with high genetic risk, the HRs were 0.809 (95% CI: 0.746-0.877) and 0.818 (95% CI: 0.754-0.888), respectively. A significant interaction was observed between genetic risk and physical activity. CONCLUSION Moderate or high levels of physical activity were associated with a lower risk of developing chronic obstructive pulmonary disease across all genetic risk groups, highlighting the need to tailor activity interventions for genetically susceptible individuals.
Humans ; Pulmonary Disease, Chronic Obstructive/epidemiology* ; Exercise ; Male ; Female ; Middle Aged ; Prospective Studies ; Aged ; Genetic Predisposition to Disease ; Risk Factors ; United Kingdom/epidemiology* ; Incidence ; Adult

Objective To analyze the efficacy and safety of different minimally invasive operations[endoscopic re-trogradebiliary drainage(ERBD)、endoscopic nasobiliary drainage(ENBD)、percutaneous transhepatic cholangial drainage(PTCD)] for choledocholithiasis complicated with acute cholangitis to provide reference for clinical treatment retrospectively. Methods A total of 151 patients with choledocholithiasis complicated with acute cholangitis at Department of Emergency Surgery in our hospital from January 2019 to December 2020 were included and divided into four groups based on the four treatment strategies, including non-surgical treatment. Changes in leukocyte count, bilirubin levels, and liver function before and after treatment, as well as postoperative recovery, complication rates, length of hospital stay, and prognosis were compared among patients who underwent different surgical treatments. Results There were significant improvements in leukocyte count, percentage of neutrophils, and liver function of the patients underwent ENBD or ERBD operation (P<0.05). The total bilirubin and direct bilirubin were significantly reduced after ERBD, ENBD, and PTCD operations (P<0.05). Patients undergoing ERBD, ENBD, or PTCD demonstrated faster recovery times, fewer complications, shorter hospital stays, and lower mortality rates compared to those managed conservatively. Conclusions ERBD and ENBD as minimally invasive therapeutic modalities for the management of choledocholithiasis complicated with acute cholangitis, exhibit remarkable clinical efficacy, coupled with a high degree of safety and reliability. These techniques significantly enhance the long-term minimally invasive cure rate, thereby establishing them as the preferred treatment strategies. Tailored to the patient's specific clinical conditions, such as the severity of infection, stone dimensions, and the use of oral anticoagulant therapy, clinicians can formulate individualized minimally invasive treatment strategies, facilitating the optimal attainment of therapeutic objectives.

Objective To explore the relationships between social support,positive coping,caregiver burden and family resilience of primary caregivers of first-stroke patients.Methods A questionnaire survey was conducted among 307 primary caregivers of first-stroke patients in 2 tertiary hospitals in Shanghai by convenience sampling method.Social support rating scale,simplified coping style questionnaire,Zarit caregiver burden interview,and family resilience assessment scale were used for questionnaire survey.Pearson correlation analysis and structural equation models were used for data analysis.Results A total of 288 valid questionnaires were collected,and the effective recovery rate was 93.81％.Pearson correlation analysis showed that there was a positive correlation between family resilience and social support,family resilience and positive coping,and social support and positive coping(r=0.375,0.627,and 0.277;all P＜0.01),while caregiver burden and social support,caregiver burden and positive coping,and family resilience and caregiver burden were all negatively correlated(r=-0.203,-0.343,and-0.444;all P＜0.01).The mediating effect model was constructed with positive coping and caregiver burden as mediating variables,social support as independent variables,and family resilience as dependent variables.The results showed that social support could mediate family resilience through positive coping,with a mediating effect of 0.164,accounting for 26.1％of the total effect;social support could also affect the family resilience of the primary caregivers of first-stroke patients through the partial chain mediating effect of positive coping and caregiver burden,with a mediating effect value of 0.032,accounting for 5.1％of the total effect.Conclusion Social support can predict family resilience among primary caregivers of first-stroke patients,and positive coping and caregiver burden play chain mediating roles in the impact of social support on family resilience.

Objective To explore the relationship between axillary artery and axillary vein by combining computed tomography angiography computed tomography angiography(CTA)and computed tomography venography(CTV),and to provide imaging data for establishing an effective blood circulation pathway for vascular injury in clinical transaxillary surgery.Methods The image data of 30 patients who underwent left upper limb and axillary CTA and CTV at the same time were collected.After three-dimensional reconstruction of the images by GE AW4.6 workstation,the course,branch type and variation of axillary artery,the course of axillary vein,the reflux of subordinate branches and the relationship between axillary artery and axillary vein were observed.The length of the whole segment,the length of segment and the inner diameter of the starting point of each segment were measured and statistically analyzed.Results According to the number of branches from the main trunk,the axillary artery was divided into 7 types.According to the variation of the number of blood vessels,the axillary vein was divided into 5 types;The unknown vein branches converge into 32 branches,of which the first segment accounted for 37.5％,the second segment accounted for 46.9％,and the third segment accounted for 15.6％.According to the absence of arterial branches,the relationship between axillary artery and axillary vein was divided into 2 types.Conclusion There are many types of branches of axillary arteries and branches of axillary veins,and the variation types are complex.The changes of branches of axillary arteries affect the distribution of branches of axillary veins.Combined with CTA and CTV images,the relationship between axillary vessels can be reflected clearly and intuitively,which can provide imaging reference for the establishment of new ways of blood supply and reflux in clinical axillary treatment.

Objective To develop a method of combining electroencephalogram(EEG)/electromyography(EMG)with multi-regional fiber photometry recording to simultaneously capture the changes of neuronal activity in the whole brain and specific brain regions during epileptic seizures.Methods The mouse head was divided into left and right regions based on the middle suture of the skull.EEG electrodes(EEG/EMG)were implanted in one side,while optical fibers were implanted in the striatum,hippocampus,entorhinal cortex,and thalamus on the contralateral side to simultaneously monitor EEG,EMG,and calcium signal dynamics.Results By combining EEG/EMG with multi-regional fiber photometry recording,differences in neuronal activity across brain regions,alongside EEG and EMG,were observed during different behavioral states.In a kainic acid(KA)-induced epilepsy model,abnormal synchronous neuronal discharges in the mouse brain were accompanied by calcium signal changes in the striatum,hippocampus,entorhinal cortex,and thalamus,with the earliest changes occurring in the hippocampus.Conclusion The combined use of EEG/EMG and multi-brain-region fiber photometry is successfully implemented in mice.This method synchronously recordes abnormal calcium signal changes across multiple brain regions,along with EEG and EMG,in the KA-induced epilepsy model.

Objective:To investigate the efficacy of a domestically developed small esophageal cooling device in implementing targeted temperature management (TTM) after resuscitation and its impact on organ injury using a porcine model of cardiac arrest and resuscitation.Methods:Thirty healthy male domestic white pigs were randomly divided into four groups using a random number table: sham (S group, n=6), normothermia (NT group, n=8), surface cooling (SC group, n=8), and esophageal cooling (EC group, n=8). The S group underwent only surgical preparation, while the other groups were subjected to 12 minutes of ventricular fibrillation followed by 6 minutes of cardiopulmonary resuscitation to establish cardiac arrest. The S and NT groups maintained a core temperature of (37.5±0.5)°C using a surface blanket. In the SC and EC groups, therapeutic hypothermia was induced post-resuscitation via surface blanket or esophageal cooling catheter to achieve a target temperature of 34°C, maintained the target temperature (34±0.5)°C for 6 hours, followed by controlled rewarming at 0.5°C/h to 37°C. Core temperature was continuously monitored for 12 hours post-resuscitation. Hemodynamic parameters, including stroke volume (SV), global ejection fraction (GEF), extravascular lung water index (ELWI), and pulmonary vascular permeability index (PVPI), were assessed using pulse indicator continuous cardiac output (PiCCO) monitoring. Serum levels of cardiac troponin I (cTnI), neuron-specific enolase (NSE), creatinine (Cr), and intestinal fatty acid-binding protein (IFABP) were measured via ELISA at 2, 6, 12, and 24 hours post-resuscitation. Neurological outcomes were evaluated at 24 hours using the neurological deficit score (NDS) and cerebral performance category (CPC). Continuous variables were analyzed using one-way ANOVA. Results:During TTM, the EC group exhibited a faster cooling rate [(1.52±0.18)°C/h vs. (0.94±0.32)°C/h, P<0.05] and shorter time to target temperature [(2.32±0.43) h vs. (3.78±0.82) h, P<0.05] compared to the SC group, with comparable maintenance and rewarming ( P>0.05). Compared to the S group, the NT, SC, and EC groups demonstrated significant post-resuscitation multi-organ injury, characterized by reduced SV and GEF, elevated ELWI and PVPI, and increased serum cTnI, NSE, Cr, IFABP, NDS, and CPC scores (all P<0.05). Relative to the NT group, the SC and EC groups showed improved SV (at 1 h post-resuscitation), GEF (at 1, 2, 4, and 6 h), ELWI (at 12 h), and reduced cTnI and NSE (at 6 h), Cr and IFABP (at 2 h), and NDS and CPC (at 24 h) (all P<0.05). Compared to the SC group, the EC group exhibited lower PVPI (at 12 h), reduced cTnI, Cr, and IFABP (at 2 h), decreased NSE (at 2, 12, and 24 h), and improved NDS (at 24 h) (all P<0.05). Conclusions:In a porcine model of cardiac arrest and resuscitation, the domestic esophageal cooling device facilitated rapid induction, stable maintenance, and controlled rewarming during TTM, outperforming traditional surface cooling. This approach demonstrated superior organ protection, warranting further investigation.