1.Effect of Acupuncture at Neiguan (PC6) on Improving Autism by Promoting Myelination Through The METTL14/m⁶A/PTEN Axis Based on “Xuanfu-Suiqiao” Theory
Wei-Li DANG ; Lü-Yuan LIANG ; Yu-Xin LI ; Zhi-Yao LI ; Sai-Dan LIU ; Jia-Lei CAO ; Rong-Ze MA ; Yun-Kai WANG ; Xiao-Qing YANG ; Bing-Qi WEI ; Bing-Xiang MA
Progress in Biochemistry and Biophysics 2026;53(5):1165-1177
ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m6A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m6A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m6A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.
2.Effect of Acupuncture at Neiguan (PC6) on Improving Autism by Promoting Myelination Through The METTL14/m⁶A/PTEN Axis Based on “Xuanfu-Suiqiao” Theory
Wei-Li DANG ; Lü-Yuan LIANG ; Yu-Xin LI ; Zhi-Yao LI ; Sai-Dan LIU ; Jia-Lei CAO ; Rong-Ze MA ; Yun-Kai WANG ; Xiao-Qing YANG ; Bing-Qi WEI ; Bing-Xiang MA
Progress in Biochemistry and Biophysics 2026;53(5):1165-1177
ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m6A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m6A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m6A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.
3.Study on The Anti-aging Effects of Longevity-enriched Metabolite Dimethylglycine
Jie HU ; Gong-Yu PU ; Jun-Lin LI ; Ju CAO ; Zhi-Xin LIN ; Wei-Wei AN ; Xue-Meng LI ; Jing AN
Progress in Biochemistry and Biophysics 2026;53(4):1048-1061
ObjectiveThe exacerbating trend of global population aging poses profound socioeconomic and public health challenges, making the comprehensive elucidation of biological aging mechanisms and the discovery of effective anti-aging interventions an urgent priority in the life sciences. Based on our previous serum metabolomics findings that dimethylglycine, an intermediate metabolite of amino acid metabolism naturally present in the human body, was significantly enriched in the serum of longevity families, this study aimed to systematically investigate the anti-aging effects of dimethylglycine both in living organisms and in controlled laboratory environments, and to preliminarily elucidate its underlying molecular mechanisms. While existing literature indicates that dimethylglycine possesses antioxidant and immunomodulatory properties, its direct anti-aging efficacy and the specific molecular pathways through which it operates remain largely unexplored. MethodsTo comprehensively evaluate the anti-aging properties of dimethylglycine, we utilized replicative senescent human embryonic lung fibroblasts, specifically the WI-38 cell line, as an experimental model in a controlled laboratory environment. Cell viability and safety were thoroughly assessed using Cell Counting Kit-8 and lactate dehydrogenase release assays across various concentrations of dimethylglycine. The impact of dimethylglycine on cellular senescence phenotypes, oxidative stress, and proliferative capacity was evaluated via senescence-associated beta-galactosidase staining, reactive oxygen species fluorescence detection, and 5-ethynyl-2'-deoxyuridine incorporation assays. Furthermore, the molecular alterations of senescence-associated secretory phenotype factors and core senescence signaling pathways were quantified using quantitative reverse transcription polymerase chain reaction for the messenger RNA levels of interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1, and enzyme-linked immunosorbent assay for the measurement of p16 and p21 protein expression levels. For the living organism model, the wild-type nematode Caenorhabditis elegans was used to evaluate systemic physiological effects. We conducted a comprehensive lifespan analysis at 20°C, heat stress resistance survival assays at 35℃, senescence-associated beta-galactosidase staining, lipofuscin accumulation tracking, intracellular reactive oxygen species measurement, and Oil Red O staining to ascertain systemic lipid accumulation. Additionally, network pharmacology bioinformatics tools, including PharmMapper and STRING databases, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were utilized to predict target pathways, alongside highly detailed molecular docking simulations utilizing SwissDock and Protein-Ligand Interaction Profiler to examine interactions with the cytochrome P450 family 2 subfamily C member 9 protein. ResultsThe experimental outcomes robustly demonstrate the potent anti-aging capabilities of dimethylglycine. At the cellular level, toxicity analyses firmly confirmed that dimethylglycine is highly safe; continuous treatment with 50 mol/L and 70 mol/L of dimethylglycine for 5 d did not induce any cellular membrane damage or cytotoxicity, but rather actively promoted cellular proliferation. Utilizing the optimal standardized concentration of 50 mol/L, dimethylglycine treatment significantly ameliorated senescent phenotypic markers in human embryonic lung fibroblasts, which was evidenced by a drastic and highly significant reduction in the senescence-associated beta-galactosidase positive cell percentage (P<0.000 1) and intracellular reactive oxygen species levels (P<0.000 1), alongside a marked increase in the 5-ethynyl-2'-deoxyuridine-positive proliferation rate (P=0.003 5). On a molecular expression scale, dimethylglycine significantly downregulated the messenger RNA expression of multiple core senescence-associated secretory phenotype inflammatory factors, including interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1. Concurrently, it effectively suppressed the protein expression of critical cell cycle arrest markers, diminishing p16 protein levels by 57.3% (P=0.000 4) and p21 protein levels by 27.2% (P=0.000 7). In the nematode Caenorhabditis elegans animal model, dimethylglycine significantly extended the mean lifespan from 20.402 d to an impressive 23.066 d (P<0.000 1) and notably enhanced overall survival rates under severe heat stress environmental conditions (P=0.017). Furthermore, systemic dimethylglycine intervention significantly mitigated age-related physiological decline by decreasing bodily lipofuscin accumulation (P<0.000 1), significantly reducing senescence-associated beta-galactosidase activity, lowering systemic reactive oxygen species fluorescence (P=0.008), and effectively alleviating overall fat accumulation (P<0.000 1). Mechanistically, extensive network pharmacology and Kyoto Encyclopedia of Genes and Genomes analyses strongly revealed that the potential targets of dimethylglycine are significantly enriched in fundamental drug metabolism and oxidative stress response pathways. Precision molecular docking simulations conclusively demonstrated that dimethylglycine forms highly stable structural interactions with the cytochrome P450 family 2 subfamily C member 9 protein, specifically highlighting the definitive formation of 5 stable hydrogen bonds involving serine 365, leucine 366, and serine 429 residues, as well as two critical salt bridge formations with arginine 97 and histidine 368 residues. It is additionally predicted to interact favorably with glutathione S-transferase family proteins. ConclusionDimethylglycine exhibits a profoundly significant and multifaceted anti-aging activity at both the cellular and entire living animal levels. By powerfully alleviating oxidative stress, heavily suppressing the core p16 and p21-dependent cellular senescence signaling pathways, and substantially mitigating the detrimental senescence-associated secretory phenotype, dimethylglycine effectively delays fundamental cellular senescence processes and drastically extends whole-organism lifespan. The biological mechanisms driving these robust protective effects are highly likely closely associated with its direct stable interactions with crucial metabolic and detoxifying enzyme systems, such as cytochrome P450 family 2 subfamily C member 9 and glutathione S-transferase family proteins, thereby systemically improving metabolic dysregulation and restoring critical redox homeostasis. This comprehensive study provides highly solid experimental evidence supporting dimethylglycine as a highly potent and safe potential anti-aging intervention agent, while simultaneously offering a clear molecular mechanistic explanation for the previously documented high abundance of dimethylglycine observed within exceptionally long-lived human populations.
4.Improvements and Recent Advances of Metadynamics Enhanced Sampling Method
Ming-Qiong TONG ; Yue-Wen YIN ; Zhi-Hong SHI ; Zan-Xia CAO
Progress in Biochemistry and Biophysics 2026;53(6):1793-1797
The functional realization of proteins and other biological macromolecules depends on conformational dynamics and allosteric regulation, and elucidating their molecular mechanisms is an important foundation for understanding life processes. Molecular dynamics simulations are a powerful tool for investigating conformational evolution at the atomic level. However, traditional methods are limited by simulation timescales and high free-energy barriers, making it difficult to effectively capture rare conformations and their transition pathways. As a result, the development of enhanced sampling techniques has become key to overcoming this bottleneck. As a classical enhanced sampling technique, metadynamics suffers from several shortcomings, including strong dependence on collective variables and significant errors caused by bias potential accumulation. This article reviews three major improvement strategies. The first combines stochastic resetting with metadynamics, using trajectory-resetting mechanisms to improve sampling efficiency while avoiding the difficulty of optimizing collective variables. The second, SinkMeta, employs a “sinking” bias effect to enable efficient exploration of specific regions and paths. The third, OPES-based hybrid methods, improve the stability of free-energy estimation by optimizing the target distribution or the way the bias is constructed. These methods provide new ideas for characterizing free-energy landscapes and studying conformational transitions in complex biological systems, while also promoting the continued development of enhanced sampling methodologies.
5.Efficacy of acupuncture combined with intermittent catheterisation in the treatment of urinary retention after radical cervical cancer surgery:a single-arm retrospective study
Ya-lin YUE ; Jing ZHU ; Yi-sheng WANG ; Jing LI ; Yun WANG ; Qi CAO ; Zhi-ling ZHU
Fudan University Journal of Medical Sciences 2025;52(5):694-700
Objective To analyse the clinical efficacy and safety of acupuncture combined with intermittent catheterisation in treating postoperative urinary retention after radical surgery for cervical cancer,and provide reference for the clinical treatment of postoperative urinary retention.Methods A retrospective analysis was conducted based on the clinical data of patients who underwent radical hysterectomy for cervical cancer at Obstetrics and Gynecology Hospital,Fudan University,from Sept 2019 to Dec 2021 and subsequently developed urinary retention.The patients were treated with a combination of acupuncture and intermittent catheterization.We observed residual urine volume,duration of acupuncture treatment,and urinary tract infection status to evaluate the effectiveness and safety of the acupuncture combined with intermittent catheterization regimen.We examined the relationship between treatment duration and various factors such as age,pathological type,surgical-pathological staging,surgical method,pre-treatment catheter indwelling time,pre-treatment residual urine volume,presence or absence of postoperative radiotherapy and chemotherapy,pre-existing urinary tract infection,and whether ureteral stenting was performed,in order to analyze the factors affecting the treatment duration.Results A total of 156 patients were reviewed.After treatment,132 cases had a post-treatment residual urine volume<100 mL,7 cases had a volume of 100-200 mL,and 15 cases had a volume>200 mL.Two cases still had great difficulty in urination after treatment,resulting in an overall effectiveness rate of 89.1%.No significant adverse events occurred in the patients.The treatment duration ranged from 2 to 17 days,with an average treatment time of 6.2 days.The treatment duration was a correlated with the preoperative indwelling catheter duration,residual urine volume and postoperative radiotherapy.Conclusion Acupuncture combined with intermittent catheterisation in treating urinary retention after radical surgery for cervical cancer has good efficacy,short treatment time and no obvious adverse effects,and can be an option for treating postoperative urinary retention.
6.Development and validation of a predictive model for postoperative blood pressure outcomes in primary aldosteronism based on CYP11B2 gene polymorphism
Qiangfeng FU ; Yongjia CHEN ; Shengtao ZENG ; Haoxiang XU ; Chenglin YANG ; Yue YANG ; Zhi CAO ; Wei WANG
Chinese Journal of Urology 2025;46(7):529-536
Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females,with a body mass index(BMI)of(25.50 ± 2.03)kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery,and polymerase chain reaction-restriction fragment length polymorphisms(PCR-RFLP)were used to detect CYP11B2(rs1799998)promoter region 344(C > T)base substitution. The follow-up duration was more than 6 months,with the following parameters recorded at the last follow-up:plasma aldosterone,renin,serum potassium,and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score(PASO)for primary aldosteronism,and the specific criteria were as follows. ① Clinical complete remission:the patient's blood pressure returned to normal(< 140/90 mmHg,1 mmHg = 0.133 kPa)and all antihypertensive drugs were discontinued;②Partial clinical remission:blood pressure returns to normal,and the number or dose of antihypertensive drugs is reduced compared with before;③Clinical non-remission:blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis,followed by binary logistic forward conditional regression for multivariate analysis,and a variety of machine learning algorithms such as random forest,logistic regression,support vector machine and gradient lifter were integrated,and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model,and the predictive performance of the model was evaluated by using receiver operating characteristic(ROC)curve,calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2(344C > T)(rs1799998)as follows:CC type in 50 cases(43.1%),CT type in 46 cases(39.7%),and TT type in 20 cases(17.3%). There were 74 cases in the complete remission group and 42 cases in the incomplete remission group,and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index[(24.27 ± 2.90)kg/m 2 vs.(26.98 ± 3.17)kg/m 2, P<0.001],preoperative hypertension grade(grade 1/2/3:29/29/16 cases vs. 9/13/20 cases, P = 0.012),preoperative antihypertensive drugs(0/1/≥ 2:25/32/17 cases vs. 7/15/20 cases, P = 0.016),and CYP11B2(344C > T)(CC/TT + CT:39/35 cases vs. 11/31 cases, P = 0.006)were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs[≥ 2: OR = 5.26(95% CI 1.12?24.61, P = 0.016;1: OR = 4.55(95% CI 1.23?22.47), P = 0.025]was the strongest independent predictor,followed by CYP11B2(344C > T)[ OR = 4.02(95% CI 1.16?13.82), P = 0.028]and BMI[ OR = 3.96(95% CI 2.26?6.92), P < 0.001]. Comparing the receiver operating feature(ROC)curves of the four types of machine learning models,the best model was the support vector machine model with an area under the curve(AUC)of 0.88(95% CI 0.82?0.95),followed by the gradient elevator model of 0.83(95% CI 0.76?0.91),the logistic regression model of 0.78(95% CI 0.68?0.88),and the random forest model of 0.77(95% CI 0.68?0.86). The optimal threshold of the Yoden index of the support vector machine model was 0.588,with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms,BMI,and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients,providing new evidence for personalized treatment strategies
7.Design and application of novel protective ventilator circuit component
Wei-zhou WU ; Kang LU ; Jing-jie CAO ; Zhi-hua ZHAO ; Hai-tao LAN ; Zan-chao CHEN ; Qing-feng XUE
Chinese Medical Equipment Journal 2025;46(4):113-117
Objective To develop a novel protective ventilator circuit component and to verify its performance by water seal and anti-splash experiments.Methods A novel protective ventilator circuit component had a design scheme with the multifunctional joint,and consisted of a tee connection tube,an isolation sleeve and a stop sleeve,of which,the tee connection tube was made of polyethylene polymer material and the others were made of silicone material.The tee connection tube had a T-shaped structure with two standard connection ports,which was composed of an adapter,a sealing cap,a plug and a sealing ring;the isolation sleeve was in the shape of a cylinder with a raised bottom,which was inserted into the adapter;the stop sleeve was located in the isolation sleeve,with an inverted frustum of a cone at the bottom and a rounded hole in the middle of the inverted frustum.An open ventilator circuit tube was involved in the performance verification of the circuit component developed.In the water seal experiment,sputum aspiration was simulated and the heights of the liquid level drop in the L-shaped tubes were compared after sputum aspiration.In the anti-splash experiment,the infection rates on the surfaces of the sterile hole towels and gloves were calculated.Results Water seal experiment showed after sputum aspiration the open ventilator circuit tube had the liquid level at the L-shaped tube higher significantly than that of the circuit component;the anti-splash experiment indicated sputum aspiration resulted in the occurance of the splashing out of the secretion and 77.5%infection rate by the open ventilator circuit tube,while no splashing out and 0%infection rate by the circuit component developed.Conclusion The novel protective ventilator circuit component behaves well in sealing and anti-splashing,and thus is worthy of clinical application for sputum aspiration.[Chinese Medical Equipment Journal,2025,46(4):113-117]
8.Effects of Liangxue Heying Formula-medicated serum on the activation of LPS-induced human umbilical vein endothelial cells via JAK2/STAT3 signaling pathway
Chun-lan XIA ; Xia FENG ; Ye-min CAO ; Zhi-qiang LIANG ; Yi LE ; Hong-tao XU ; Fei QI ; Ji LI
Chinese Traditional Patent Medicine 2025;47(4):1150-1156
AIM To investigate the effects of Liangxue Heying Formula-medicated serum(LXHY-MS)on human umbilical vein endothelial cells(HUVECs)induced by lipopolysaccharide(LPS).METHODS CCK-8,DCFH-DA fluorescence probe and Western blot method were used to screen the LPS concentration in modeling and the serum LXHY concentration for treatment.The HUVECs divided into the normal group,the model group and the LXHY-MS group had their SOD activity detected by automatic biochemical analyzer;their MDA level detected by colorimetry;their protein expressions of ICAM-1,VCAM-1,IL-6,TNF-α,p-JAK2 and p-STAT3 detected by Western blot;and their mROS expression and recruitment effect on THP-1 photographed with high connotation.With the use of JAK2/STAT3 pathway inhibitor(AG490),the HUVECs divided into the normal group,the AG490 group,the LPS group,the LPS+AG490 group,the LPS+LXHY-MS group,and LPS+LXHY-MS+AG490 group were subjected to the corresponding treatment,followed by the detection of their protein expressions of ICAM-1,VCAM-1,IL-6,TNF-α,p-JAK2 and p-STAT3 by Western blot.RESULTS Compared with the normal group,the model group displayed decreased SOD activity(P<0.01),increased MDA level(P<0.05),increased ICAM-1,VCAM-1,IL-6,TNF-α,p-JAK2,p-STAT3 protein expressions(P<0.05,P<0.01),and increased mROS expression and THP-1 cells recruitment.Compared with the model group,the LXHY-MS group shared increased SOD activity(P<0.05),decreased MDA level(P<0.01),decreased ICAM-1,VCAM-1,IL-6,TNF-α,p-JAK2,p-STAT3 protein expressions(P<0.05,P<0.01),reduced mROS expression and THP-1 cells recruitment.Given the use of AG490,the model group displayed increased protein expressions of ICAM-1,VCAM-1,IL-6,TNF-α,p-JAK2 and p-STAT3 in contrast to the normal group(P<0.05,P<0.01);each intervened group showed decreased expressions of related proteins in contrast to the model group(P<0.05,P<0.01).CONCLUSION LXHY-MS may protect the injury due to the activation of HUVECs by inhibiting the JAK2/STAT3 signaling pathway.
9.Therapeutic mechanism of Compound Xuanju Capsule on erectile dysfunction
Zi-jie LI ; Hao-xiang XU ; Wei WANG ; Yue YANG ; Cheng-lin YANG ; Zhi CAO ; Xiao-ming ZHANG
National Journal of Andrology 2025;31(8):675-683
Objective:To investigate the pharmacological mechanism of Compound Xuanju Capsule in the treatment of erectile dysfunction(ED)by using network pharmacology and molecular docking technology.Methods:The active ingredients and targets of Compound Xuanju Capsule were screened using Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform(TCMSP).TTD,OMIM,DrugBank and GeneCards databases were used to obtain genes related to ED,and the union of the results was taken as the disease genes of ED.The common target of drug and disease was taken as the potential target of Compound Xuanju Capsule in ED,and the drug-disease interaction network was constructed by using Cytoscape software.The protein-protein interaction(PPI)network was constructed by using String database,which was then imported into Cytoscape to identify the key target.Based on the drug-disease intersection genes,GO and KEGG enrichment analyses were performed to predict the relevant signaling pathways and molecular mechanisms of Compound Xuanju Capsule for the treatment of ED.Autodock software was used to perform molecular docking between the active ingredients and the core targets.Results:Forty chemical components of Compound Xuanju Capsule were screened,and 239 predicted targets were obtained.A total of 1 907 ED-related genes were screened,and 97 common targets were identified between Compound Xuanju Capsule and ED,among which the core targets included EGFR,ESR1,HIF1A,PTGS2,and STAT3.The signaling pathways obtained by KEGG enrichment analysis included calcium signaling pathway,HIF-1 signaling pathway,PI3K-Akt signaling pathway,cGMP-PKG signaling pathway,relaxin signaling pathway,Serotonergic synapse signaling pathway.The molecular docking results showed that there were molecular binding sites between the key active ingredients and the core targets with strong binding activity.Conclusion:Compound Xuanju Capsule may treat ED through multi-target pathways such as anti-inflamnmato-ry and improving cellular oxidative stress.
10.Development and validation of a predictive model for postoperative blood pressure outcomes in primary aldosteronism based on CYP11B2 gene polymorphism
Qiangfeng FU ; Yongjia CHEN ; Shengtao ZENG ; Haoxiang XU ; Chenglin YANG ; Yue YANG ; Zhi CAO ; Wei WANG
Chinese Journal of Urology 2025;46(7):529-536
Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females,with a body mass index(BMI)of(25.50 ± 2.03)kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery,and polymerase chain reaction-restriction fragment length polymorphisms(PCR-RFLP)were used to detect CYP11B2(rs1799998)promoter region 344(C > T)base substitution. The follow-up duration was more than 6 months,with the following parameters recorded at the last follow-up:plasma aldosterone,renin,serum potassium,and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score(PASO)for primary aldosteronism,and the specific criteria were as follows. ① Clinical complete remission:the patient's blood pressure returned to normal(< 140/90 mmHg,1 mmHg = 0.133 kPa)and all antihypertensive drugs were discontinued;②Partial clinical remission:blood pressure returns to normal,and the number or dose of antihypertensive drugs is reduced compared with before;③Clinical non-remission:blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis,followed by binary logistic forward conditional regression for multivariate analysis,and a variety of machine learning algorithms such as random forest,logistic regression,support vector machine and gradient lifter were integrated,and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model,and the predictive performance of the model was evaluated by using receiver operating characteristic(ROC)curve,calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2(344C > T)(rs1799998)as follows:CC type in 50 cases(43.1%),CT type in 46 cases(39.7%),and TT type in 20 cases(17.3%). There were 74 cases in the complete remission group and 42 cases in the incomplete remission group,and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index[(24.27 ± 2.90)kg/m 2 vs.(26.98 ± 3.17)kg/m 2, P<0.001],preoperative hypertension grade(grade 1/2/3:29/29/16 cases vs. 9/13/20 cases, P = 0.012),preoperative antihypertensive drugs(0/1/≥ 2:25/32/17 cases vs. 7/15/20 cases, P = 0.016),and CYP11B2(344C > T)(CC/TT + CT:39/35 cases vs. 11/31 cases, P = 0.006)were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs[≥ 2: OR = 5.26(95% CI 1.12?24.61, P = 0.016;1: OR = 4.55(95% CI 1.23?22.47), P = 0.025]was the strongest independent predictor,followed by CYP11B2(344C > T)[ OR = 4.02(95% CI 1.16?13.82), P = 0.028]and BMI[ OR = 3.96(95% CI 2.26?6.92), P < 0.001]. Comparing the receiver operating feature(ROC)curves of the four types of machine learning models,the best model was the support vector machine model with an area under the curve(AUC)of 0.88(95% CI 0.82?0.95),followed by the gradient elevator model of 0.83(95% CI 0.76?0.91),the logistic regression model of 0.78(95% CI 0.68?0.88),and the random forest model of 0.77(95% CI 0.68?0.86). The optimal threshold of the Yoden index of the support vector machine model was 0.588,with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms,BMI,and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients,providing new evidence for personalized treatment strategies

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