Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.

ObjectiveTo examine the influences of Shenfu injection on the endogenous metabolic byproducts in the myocardium of the rat model exhibiting chronic heart failure， thus deciphering the therapeutic mechanism of the Qi-reinforcing and Yang-warming method. MethodsSD rats were randomly allocated into a control group and a modeling group. Chronic heart failure with heart-Yang deficiency syndrome in rats was modeled by multi-point subcutaneous injection of isoproterenol， and the rats were fed for 14 days after modeling. The successfully modeled rats were randomized into model， Shenfu injection （6.0 mL·kg^-1）， and trimetazidine （10 mg·kg^-1） groups and treated with corresponding agents for 15 days. The control group and the model group were injected with equal doses of normal saline， and the samples were collected after the intervention was completed. Cardiac color ultrasound was performed. Hematoxylin-eosin （HE） staining was used to observe histopathological morphology， and the serum level of N-terminal pro-brain natriuretic peptide （NT-proBNP） was assessed by enzyme-linked immunosorbent assay （ELISA）. The mitochondrial morphological and structural changes of cardiomyocytes were observed by transmission electron microscopy， and the metabolic profiling was carried out by ultra high performance liquid chromatography-quantitative exactive-mass spectrometry （UHPLC-QE-MS）. Differential metabolites were screened and identified by orthogonal partial least squares-discriminant analysis （OPLS-DA） and other methods， and then the MetaboAnalyst database was used for further screening. The relevant biological pathways were obtained through pathway enrichment analysis. The receiver operating characteristic （ROC） curve was established to evaluate the diagnostic value of each potential biomarker for myocardial injury and the evaluation value for drug efficacy. ResultsThe results of color ultrasound showed that Shenfu Injection improved the cardiac function indexes of model rats （P<0.05）. The results of HE staining showed that Shenfu injection effectively alleviated the pathological phenomena such as myocardial tissue structure disorder and inflammatory cell infiltration in model rats. The results of ELISA showed that Shenfu injection effectively regulated the serum NT-proBNP level in the model rats. Transmission electron microscopy （TEM） showed that Shenfu injection effectively restored the mitochondrial morphological structure. The results of metabolomics showed that the metabolic phenotypes of myocardial samples presented markedly differences between groups. Nine differential metabolites could be significantly reversed in the Shenfu injection group， involving three metabolic pathways： pyruvate metabolism， histidine metabolism， and citric acid cycle （TCA cycle）. The results of ROC analysis showed that the area under the curve （AUC） values of all metabolites were between 0.75 and 1.0， indicating that the differential metabolites had high diagnostic accuracy for myocardial injury， and the changes in their expression levels could be used as potential markers for efficacy evaluation. ConclusionShenfu injection significantly alleviated the damage of cardiac function， myocardium， and mitochondrial structure in the rat model of chronic heart failure with heart-Yang deficiency syndrome by ameliorating energy metabolism remodeling. Reinforcing Qi and warming Yang is a key method for treating chronic heart failure with heart-Yang deficiency syndrome.

ObjectiveTo investigate the mechanism of action of remifentanil （RMZL） in alleviating lung ischemia-reperfusion injury （LIRI） in rats by inhibiting pyroptosis through modulating hypoxia inducible factor-1α （HIF-1α）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3） pathway. MethodsRats were stochastically assigned into Control group， LIRI group， RMZL low-dose group， RMZL medium-dose group， RMZL high-dose group， and RMZL high-dose+HIF-1α activator dimethyloxallyl glycine （DMOG） group， with 18 rats in each group. Rats in Control group only had their left pulmonary hilum free and did not undergo ischemia-reperfusion treatment. Except for the Control group， LIRI models were constructed in all other groups. Rats in LIRI group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before constructing LIRI model； rats in Control group were intraperitoneally injected with an equal amount of physiological saline 15 minutes before freeing left pulmonary hilum； rats in other groups were intraperitoneally injected with corresponding dose of drug 15 minutes before constructing LIRI model. The wet/dry weight ratio of lungs was calculated. HE staining was used to study lung tissue pathology. Immunofluorescence staining was used to detect the relative fluorescence intensity of gasdermin D （GSDMD） and NLRP3 double positive cells in lung tissue. ELISA was used to detect interleukin-1β and IL-18 in lung tissue. Western blot was used to detect HIF-1α， NLRP3， cysteine-aspartic protease-1 （Cleaved caspase-1）， and gasdermin D-N （GSDMD-N） proteins in lung tissue. ResultsCompared to the Control group， the LIRI group showed disordered alveolar structure， thickened alveolar septa， and abundant inflammatory cell infiltration in rats. The lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， IL-1β， IL-18 levels， and HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N proteins increased （P0.05）. For the LIRI group， rats in the RMZL low， medium， and high-dose groups displayed attenuated alveolar septal thickening and reduced inflammatory cell infiltration. The lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， IL-1β， IL-18 levels， and HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N proteins declined， and the RMZL high-dose group showed the most prominent trend （P0.05）. Compared with the RMZL high-dose group， rats in the RMZL high-dose+DMOG group exhibited thickened alveolar septa and more inflammatory cell infiltration， along with increased lung wet/dry weight ratio， relative fluorescence intensity of GSDMD and NLRP3 double positive cells in lung tissue， levels of IL-1β and IL-18， and protein expression of HIF-1α， NLRP3， Cleaved caspase-1， and GSDMD-N （P0.05）. ConclusionRMZL may inhibit pyroptosis in LIRI rats by suppressing HIF-1α/NLRP3 pathway.

ObjectiveTo investigate whether there exists an independent association beyond shared risk factors between colorectal space-occupying lesions and metabolic associated fatty liver disease （MAFLD）， and to provide new ideas for early identification of colorectal space-occupying lesions in the MAFLD population. MethodsA retrospective analysis was performed for 12 252 patients who were hospitalized in The First Affiliated Hospital of Shandong First Medical University （Shandong Provincial Qianfoshan Hospital） and underwent both colonoscopy and abdominal imaging （abdominal ultrasound， magnetic resonance imaging， or computed tomography） from September 1， 2018 to August 31， 2023. According to colonoscopy findings， the patients were divided into colorectal space-occupying lesion group with 6 545 patients and non-colorectal space-occupying lesion group with 5 707 patients. Baseline data were compared between the two groups. The univariate and multivariate Logistic regression analyses were used to identify influencing factors for colorectal space-occupying lesions， and further analysis was performed to investigate the association of MAFLD and liver fibrosis （assessed by fibrosis-4 ［FIB-4］， nonalcoholic fatty liver disease fibrosis score ［NFS］， and aspartate aminotransferase-to-platelet ratio index） with colorectal space-occupying lesions. The Mann-Whitney U test was used for comparison of continuous data with skewed distribution between groups， and the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. ResultsThe univariate analysis showed that compared with the non-colorectal space-occupying lesion group， the colorectal space-occupying lesion group had significantly higher age （Z=-95.628， P<0.001）， proportion of male patients （χ²=406.910， P<0.001）， body weight （Z=-24.928， P<0.001）， body mass index （BMI） （Z=-21.367， P<0.001）， diastolic blood pressure （Z=-15.527， P<0.001）， systolic blood pressure （Z=-16.965， P<0.001）， proportion of patients with history of coronary heart disease （χ²=28.112， P<0.001）/hypertension （χ²=152.993， P<0.001）/diabetes （χ²=52.175， P<0.001）/cerebral infarction （χ²=14.097， P<0.001）， white blood cell count （WBC） （Z=-12.801， P<0.001）， hemoglobin （HGB） （Z=-19.258， P<0.001）， aspartate aminotransferase （Z=-6.673， P=0.001）， alanine aminotransferase （Z=-10.375， P<0.001）， alkaline phosphatase （Z=-10.496， P<0.001）， gamma-glutamyl transpeptidase （GGT） （Z=-18.893， P<0.001）， blood urea nitrogen （Z=-13.291， P<0.001）， creatinine （Z=-16.798， P<0.001）， uric acid （Z=-16.822， P<0.001）， triglyceride （Z=-10.186， P<0.001）， and fasting blood glucose （Z=-15.761， P<0.001）. The multivariate Logistic regression analysis showed that age （odds ratio ［OR］=1.052， 95% confidence interval ［CI］： 1.045 — 1.057， P<0.001）， sex （OR=0.523， 95%CI： 0.466 — 0.587， P<0.001）， BMI （OR=1.031， 95%CI： 1.018 — 1.046， P<0.001）， history of hypertension （OR=1.140， 95%CI： 1.019 — 1.276， P=0.022）， WBC （OR=1.057， 95%CI： 1.028 — 1.087， P<0.001）， GGT （OR=1.001， 95%CI： 1.000 — 1.002， P=0.021）， Alb （OR=0.982， 95%CI： 0.969 — 0.995， P=0.008）， and HGB （OR=1.004， 95%CI： 1.001 — 1.008， P=0.019） were independent influencing factors for colorectal space-occupying lesions. In the MAFLD population， after adjustment for age， sex， BMI， history of hypertension， WBC， HGB， GGT， and Alb， intermediate-to-high risk of advanced liver fibrosis assessed by FIB-4 and NFS was still an influencing factor for colorectal space-occupying lesions （FIB-4： OR=1.457， 95%CI： 1.176 — 1.810， P<0.05； NFS： OR=1.499， 95%CI： 1.244 — 1.809， P<0.05）. Liver fibrosis degree based on FIB-4 was not significantly associated with the pathological type， location， size， or number of colorectal space-occupying lesions （all P>0.05）. While intermediate-to-high risk of advanced liver fibrosis based on NFS was not associated with pathological type， location， or size， it might be associated with the number of lesions （χ²=9.770， P<0.05）. ConclusionAge， sex， BMI， history of hypertension， WBC， HGB， GGT， and Alb are independent influencing factors for colorectal space-occupying lesions. Intermediate-to-high risk of advanced liver fibrosis assessed by FIB-4 or NFS is independently associated with colorectal space-occupying lesions， and liver fibrosis assessed by NFS might be associated with the increase in the number of lesions.

ObjectiveTo measure the distance from the surface of Fengfu （GV 16） to the deep important bony structures in children aged 4 to 8 years based on CT three-dimensional reconstruction technology， thereby providing imaging reference data for clinical acupuncture safe operations in terms of different age groups and needle insertion angles. MethodsCervical CT image data were collected from healthy children aged 4 to 8 years with five age groups （4-， 5-， 6-， 7-， 8-year-old groups）， 20 cases in each group. The images were imported into Mimics 21.0 software for three-dimensional reconstruction. The straight-line distances from the skin model surface of the Fengfu （GV 16） to the first underlying bony barrier （such as the lower edge of the occipital bone， the posterior arch of the atlas vertebra） were measured at intervals of 5° on both the transverse section （0° ± 35°） and the sagittal section （40° upward to 40° downward on the diagonal）. The correlation between the measured distance， age， and angle was analyzed. ResultsIn the transverse and sagittal sections， the measured distance from the skin surface to the underlying bone of Fengfu （GV 16） at each angle increased with age. In the transverse section， the increase was relatively small within the 0° to 20° angle range， whereas a more pronounced increase was observed when the angle exceeded 20°； at measurement angles of 30° to 35°， the distance of the skin surface to the bone at Fengfu （GV 16） in the 7- and 8-year-old groups was significantly greater than that in the 4-year-old group （P＜0.05）. In the sagittal section， the measured distance from the skin surface to the bone at Fengfu （GV 16） was shortest and changed greatly at oblique upward angles of 15° to 40° within each age group， and increased markedly at near-vertical angles of 0° to 10°； it was longest at oblique downward angles of -40° to -5° and showed relatively gradual changes. ConclusionThis study obtained the imaging distance data from Fengfu （GV 16） to the surrounding bony structures in children aged 4 to 8 years through three-dimensional modeling， finding that this distance has clear age and angle dependence， which can provide quantitative evidence for developing individualized and precise safe acupuncture schemes at Fengfu （GV 16） in pediatric clinical practice.

ObjectiveTo measure the distance from the surface of Fengfu （GV 16） to the deep important bony structures in children aged 4 to 8 years based on CT three-dimensional reconstruction technology， thereby providing imaging reference data for clinical acupuncture safe operations in terms of different age groups and needle insertion angles. MethodsCervical CT image data were collected from healthy children aged 4 to 8 years with five age groups （4-， 5-， 6-， 7-， 8-year-old groups）， 20 cases in each group. The images were imported into Mimics 21.0 software for three-dimensional reconstruction. The straight-line distances from the skin model surface of the Fengfu （GV 16） to the first underlying bony barrier （such as the lower edge of the occipital bone， the posterior arch of the atlas vertebra） were measured at intervals of 5° on both the transverse section （0° ± 35°） and the sagittal section （40° upward to 40° downward on the diagonal）. The correlation between the measured distance， age， and angle was analyzed. ResultsIn the transverse and sagittal sections， the measured distance from the skin surface to the underlying bone of Fengfu （GV 16） at each angle increased with age. In the transverse section， the increase was relatively small within the 0° to 20° angle range， whereas a more pronounced increase was observed when the angle exceeded 20°； at measurement angles of 30° to 35°， the distance of the skin surface to the bone at Fengfu （GV 16） in the 7- and 8-year-old groups was significantly greater than that in the 4-year-old group （P＜0.05）. In the sagittal section， the measured distance from the skin surface to the bone at Fengfu （GV 16） was shortest and changed greatly at oblique upward angles of 15° to 40° within each age group， and increased markedly at near-vertical angles of 0° to 10°； it was longest at oblique downward angles of -40° to -5° and showed relatively gradual changes. ConclusionThis study obtained the imaging distance data from Fengfu （GV 16） to the surrounding bony structures in children aged 4 to 8 years through three-dimensional modeling， finding that this distance has clear age and angle dependence， which can provide quantitative evidence for developing individualized and precise safe acupuncture schemes at Fengfu （GV 16） in pediatric clinical practice.

Objective To analyze the status of willingness to participate in health science popularization (HSP) and its influencing factors among medical workers in Guangdong Province. Methods A total of 462 medical workers in Guangdong Province were recruited as the study subjects using a stratified convenience sampling method. The Questionnaire on Influencing Factors of Willingness to Participate in Health Science Popularization, developed by the research team, was used to investigate their willingness to participate in HSP and related variables, including previous communication experience, attitude, moral belief, descriptive norm, injunctive norm, self-efficacy, and social support. Hierarchical regression analysis was used to identify the influencing factors of willingness to participate in HSP. Results The median and 25^th and 75^th percentiles score of willingness to participate in HSP was 4.0 (3.8, 4.5), indicating a moderately high level of willingness. The HSP participation willingness score was positively correlated with scores for previous communication experience, attitude, moral belief, descriptive norm, injunctive norm, self-efficacy, and social support (Spearman correlation coefficients were 0.513, 0.384, 0.507, 0.533, 0.463, 0.556, and 0.435, respectively; all P<0.01). Hierarchical regression analysis showed that previous communication experience, moral belief, descriptive norm and self-efficacy positively predicted willingness to participate in HSP (all P<0.05). Medical workers in the non-Pearl River Delta region had lower willingness than those in the Pearl River Delta region for HSP participation (P<0.05). Conclusion There is still room for improving willingness for HSP participation among healthcare professionals in Guangdong Province. It is recommended to enhance self-efficacy through communication skills training, strengthen professional ethics education, foster a positive peer engagement atmosphere, and intensify resource support and capacity building for HSP in the non-Pearl River Delta region, to improve HSP participation willingness among medical workers in Guangdong Province.

ObjectiveTo observe the clinical efficacy and safety of Yanghe Xiaonang Decoction （阳和消囊汤， YXD） in the treatment of polycystic ovarian syndrome （PCOS）-related infertility with healthy qi deficiency and phlegm stasis syndrome. MethodsA total of 120 PCOS-related infertility participants with healthy qi deficiency and phlegm stasis syndrome were randomly divided into a treatment group and a control group in a 1∶1 ratio， with 60 cases in each group. The treatment group started taking YXD on the first day of the menstrual cycle， one dose daily， until the day of sexual intercourse when follicles matured and were released. Those without mature follicles or ovulation could take it until the 14th to 28th day. The control group was given oral letrozole at 2.5 mg daily starting on the 5th day of the menstrual cycle for 5 consecutive days. If the basal body temperature curve remained monophasic on the 28th day of the menstrual cycle， and no dominant follicle was detected by color Doppler ultrasound， estrogen plus progesterone or progesterone alone was administered for withdrawal bleeding， followed by a new treatment cycle. The treatment lasted for three consecutive menstrual cycles. Patients were followed up for 3 months upon confirmed pregnancy or after the completion of the 3rd treatment cycle. The clinical pregnancy rate， total effective rate， pregnancy rate， biochemical pregnancy rate， embryonic arrest rate， and ectopic pregnancy rate between the two groups were compared. Transvaginal color Doppler ultrasound was used for monitoring endometrial thickness， cyclic ovulation status， number of follicles between 2-9 mm， and the difference in transverse and longitudinal diameters of dominant follicles. The levels of endocrine hormone indicators including serum testosterone （T）， prolactin （PRL）， luteinizing hormone （LH）， follicle-stimulating hormone （FSH）， progesterone （P）， estradiol （E₂）， free testosterone （F-T）， androstenedione （ASD）， inhibin B （INH-B）， and anti-Müllerian hormone （AMH）， as well as the total traditional Chinese medicine （TCM） symptom scores before and after treatment were compared between the two groups. ResultsA total of 58 patients in each group were included in the statistical analysis. The clinical pregnancy rate in the treatment group was 39.66% （23/58）， higher than 22.41% （13/58） in the control group （P＜0.05）. The total effective rate in the treatment group was 96.55% （56/58）， higher than 79.31% （46/58） in the control group （P＜0.05）. The biochemical pregnancy rate and embryonic arrest rate in the treatment group were lower than those in the control group （P＜0.05）. The treatment group was superior to the control group in increasing endometrial thickness and reducing the number of 2-9 mm follicles at the 2nd and 3rd cycles， as well as in improving the cyclic ovulation status and the difference in transverse and longitudinal diameters of dominant follicles at the 3rd cycle （P＜0.05）. After treatment， the treatment group exhibited significant reductions in serum T， LH， F-T， ASD， INH-B， and AMH levels （P＜0.05）. The control group also showed significant decreases in LH， F-T， INH-B， and AMH levels （P＜0.05）. The levels of F-T， INH-B， and AMH in the treatment group after treatment were lower than those in the control group （P＜0.05）. The improvement of the total TCM symptom score in the treatment group was superior to that in the control group （P＜0.05）. ConclusionYXD can significantly improve the clinical pregnancy rate， reduce the rates of biochemical pregnancy and embryonic arrest， alleviate clinical symptoms， increase endometrial thickness， enhance dominant follicles， and improve endocrine hormone levels in PCOS-related infertility with healthy qi deficiency and phlegm stasis syndrome， showing good safety.

Nine compounds were isolated and purified from 90% ethanol extract of Alstonia mairei Lévl by using various chromatographic methods, including silica gel, SephadexLH-20, MCI Gel and ODS column chromatography, combined with semi-preparative liquid phase separation methods. Modern spectroscopic methods (1D and 2D NMR, UV, IR, MS, etc.) were used to identify the structures of the isolated compounds. They were identified as mairoside A (1), 3′,6-di-O-sinaloylsucrose (2), myristic acid (3), methyl myristate (4), ethyl myristate (5), 3,4,5-trimethoxycinnamic acid (6), 3,4,5-trimethoxybenzoic acid (7), vinoline (8), kaempferol-3-O-rutinoside (9), among which compound 1 is a new glycoside, compounds 4 and 5 are new natural products, and the nuclear magnetic data of compound 4 were reported for the first time.

Circadian rhythm is an endogenous biological clock mechanism that enables organisms to adapt to the earth’s alternation of day and night. It plays a fundamental role in regulating physiological functions and behavioral patterns, such as sleep, feeding, hormone levels and body temperature. By aligning these processes with environmental changes, circadian rhythm plays a pivotal role in maintaining homeostasis and promoting optimal health. However, modern lifestyles, characterized by irregular work schedules and pervasive exposure to artificial light, have disrupted these rhythms for many individuals. Such disruptions have been linked to a variety of health problems, including sleep disorders, metabolic syndromes, cardiovascular diseases, and immune dysfunction, underscoring the critical role of circadian rhythm in human health. Among the numerous systems influenced by circadian rhythm, the skin—a multifunctional organ and the largest by surface area—is particularly noteworthy. As the body’s first line of defense against environmental insults such as UV radiation, pollutants, and pathogens, the skin is highly affected by changes in circadian rhythm. Circadian rhythm regulates multiple skin-related processes, including cyclic changes in cell proliferation, differentiation, and apoptosis, as well as DNA repair mechanisms and antioxidant defenses. For instance, studies have shown that keratinocyte proliferation peaks during the night, coinciding with reduced environmental stress, while DNA repair mechanisms are most active during the day to counteract UV-induced damage. This temporal coordination highlights the critical role of circadian rhythms in preserving skin integrity and function. Beyond maintaining homeostasis, circadian rhythm is also pivotal in the skin’s repair and regeneration processes following injury. Skin regeneration is a complex, multi-stage process involving hemostasis, inflammation, proliferation, and remodeling, all of which are influenced by circadian regulation. Key cellular activities, such as fibroblast migration, keratinocyte activation, and extracellular matrix remodeling, are modulated by the circadian clock, ensuring that repair processes occur with optimal efficiency. Additionally, circadian rhythm regulates the secretion of cytokines and growth factors, which are critical for coordinating cellular communication and orchestrating tissue regeneration. Disruptions to these rhythms can impair the repair process, leading to delayed wound healing, increased scarring, or chronic inflammatory conditions. The aim of this review is to synthesize recent information on the interactions between circadian rhythms and skin physiology, with a particular focus on skin tissue repair and regeneration. Molecular mechanisms of circadian regulation in skin cells, including the role of core clock genes such as Clock, Bmal1, Per and Cry. These genes control the expression of downstream effectors involved in cell cycle regulation, DNA repair, oxidative stress response and inflammatory pathways. By understanding how these mechanisms operate in healthy and diseased states, we can discover new insights into the temporal dynamics of skin regeneration. In addition, by exploring the therapeutic potential of circadian biology in enhancing skin repair and regeneration, strategies such as topical medications that can be applied in a time-limited manner, phototherapy that is synchronized with circadian rhythms, and pharmacological modulation of clock genes are expected to optimize clinical outcomes. Interventions based on the skin’s natural rhythms can provide a personalized and efficient approach to promote skin regeneration and recovery. This review not only introduces the important role of circadian rhythms in skin biology, but also provides a new idea for future innovative therapies and regenerative medicine based on circadian rhythms.