1.Research progress in effect of traditional Chinese medicine on aerobic glycolysis in colorectal cancer.
Xu MA ; Sheng-Long LI ; Guang-Rong ZHENG ; Da-Cheng TIAN ; Gang-Gang LU ; Jie GAO ; Yu-Qi AN ; Li-Yuan CAO ; Liang LI ; Xiao-Yong TANG
China Journal of Chinese Materia Medica 2025;50(6):1496-1506
Colorectal cancer(CRC) is a common malignant tumor worldwide. Due to the treatment intolerance and side effects, CRC rank the top among various cancers regarding the incidence and mortality rates. Therefore, exploring new therapies is of great significance for the treatment of CRC. Aerobic glycolysis(AEG) plays an important role in the microenvironment formation, proliferation, metastasis, and recurrence of CRC and other tumor cells. It has been confirmed that intervening in the AEG pathway can effectively curb CRC. The active ingredients and compound prescriptions of traditional Chinese medicine(TCM) can effectively inhibit the proliferation, metastasis, and drug resistance and regulate the apoptosis of tumor cells by modulating AEG-associated transport proteins [eg, glucose transporters(GLUT)], key enzymes [hexokinase(HK) and phosphofructokinase(PFK)], key genes [hypoxia-inducible factor 1(HIF-1) and oncogene(c-Myc)], and signaling pathways(MET/PI3K/Akt/mTOR). Accordingly, they can treat CRC, reduce the recurrence, and improve the prognosis of CRC. Although AEG plays a key role in the development and progression of CRC, the specific mechanisms are not yet fully understood. Therefore, this article delves into the intrinsic connection of the targets and mechanisms of the AEG pathway with CRC from the perspective of tumor cell glycolysis and explores how active ingredients(oxymatrine, kaempferol, and dioscin) and compound prescriptions(Quxie Capsules, Jiedu Sangen Decoction, and Xianlian Jiedu Prescription) of TCM treat CRC by intervening in the AEG pathway. Additionally, this article explores the shortcomings in the current research, aiming to provide reliable targets and a theoretical basis for treating CRC with TCM.
Humans
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Colorectal Neoplasms/genetics*
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Drugs, Chinese Herbal/therapeutic use*
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Glycolysis/drug effects*
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Animals
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Medicine, Chinese Traditional
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Signal Transduction/drug effects*
2.Randomized, double-blind, parallel-controlled, multicenter, equivalence clinical trial of Jiuwei Xifeng Granules(Os Draconis replaced by Ostreae Concha) for treating tic disorder in children.
Qiu-Han CAI ; Cheng-Liang ZHONG ; Si-Yuan HU ; Xin-Min LI ; Zhi-Chun XU ; Hui CHEN ; Ying HUA ; Jun-Hong WANG ; Ji-Hong TANG ; Bing-Xiang MA ; Xiu-Xia WANG ; Ai-Zhen WANG ; Meng-Qing WANG ; Wei ZHANG ; Chun WANG ; Yi-Qun TENG ; Yi-Hui SHAN ; Sheng-Xuan GUO
China Journal of Chinese Materia Medica 2025;50(6):1699-1705
Jiuwei Xifeng Granules have become a Chinese patent medicine in the market. Because the formula contains Os Draconis, a top-level protected fossil of ancient organisms, the formula was to be improved by replacing Os Draconis with Ostreae Concha. To evaluate whether the improved formula has the same effectiveness and safety as the original formula, a randomized, double-blind, parallel-controlled, equivalence clinical trial was conducted. This study enrolled 288 tic disorder(TD) of children and assigned them into two groups in 1∶1. The treatment group and control group took the modified formula and original formula, respectively. The treatment lasted for 6 weeks, and follow-up visits were conducted at weeks 2, 4, and 6. The primary efficacy endpoint was the difference in Yale global tic severity scale(YGTSS)-total tic severity(TTS) score from baseline after 6 weeks of treatment. The results showed that after 6 weeks of treatment, the declines in YGTSS-TSS score showed no statistically significant difference between the two groups. The difference in YGTSS-TSS score(treatment group-control group) and the 95%CI of the full analysis set(FAS) were-0.17[-1.42, 1.08] and those of per-protocol set(PPS) were 0.29[-0.97, 1.56], which were within the equivalence boundary [-3, 3]. The equivalence test was therefore concluded. The two groups showed no significant differences in the secondary efficacy endpoints of effective rate for TD, total score and factor scores of YGTSS, clinical global impressions-severity(CGI-S) score, traditional Chinese medicine(TCM) response rate, or symptom disappearance rate, and thus a complete evidence chain with the primary outcome was formed. A total of 6 adverse reactions were reported, including 4(2.82%) cases in the treatment group and 2(1.41%) cases in the control group, which showed no statistically significant difference between the two groups. No serious suspected unexpected adverse reactions were reported, and no laboratory test results indicated serious clinically significant abnormalities. The results support the replacement of Os Draconis by Ostreae Concha in the original formula, and the efficacy and safety of the modified formula are consistent with those of the original formula.
Adolescent
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Child
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Child, Preschool
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Female
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Humans
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Male
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Double-Blind Method
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Drugs, Chinese Herbal/therapeutic use*
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Tic Disorders/drug therapy*
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Treatment Outcome
3.Structurally novel tryptamine-derived alkaloids from the seeds of Peganum harmala and their antiviral activities against respiratory syncytial virus.
Zhongnan WU ; Yubo ZHANG ; Guocai WANG ; Qing TANG ; Yaolan LI ; Xiaoqing XIE ; Yushen LIANG ; Wen CHENG
Chinese Journal of Natural Medicines (English Ed.) 2025;23(8):972-979
Peganum harmala L. (P. harmala) is a significant economic and medicinal plant. The seeds of P. harmala have been extensively utilized in traditional Chinese medicine, Uighur medicine, and Mongolian medicine, as documented in the Drug Standard of the Ministry of Health of China. Twelve novel tryptamine-derived alkaloids (1-12) and eight known compounds (13-20) were isolated from P. harmala seeds. Compounds 1 and 2 represent the first reported instances of tryptamine-derived heteromers, comprising tryptamine and aniline fragments with previously undocumented C-3-N-1' linkage and C-3-C-4' connection, respectively. Compounds 3-5 were identified as indole-quinazoline heteromers, exhibiting a novel C-3 and NH-1' linkage between indole and quinazoline-derived fragments. Compound 6 demonstrates the dimerization pattern of C-C linked tryptamine-quinazoline dimer. Compound 8 represents a tryptamine-derived heterodimer with a distinctive carbon skeleton, featuring an unusual spiro-tricyclic ring (7) and conventional bicyclic tryptamine. Compounds 9-11 constitute novel 6/5/5/5 spiro-tetracyclic tryptamine-derived alkaloids presenting a unique ring system of tryptamine-spiro-pyrrolizine. Compounds 1-3 and 6-11 were identified as racemates. Compounds 2, 7, 9, 10, and 12 were confirmed via X-ray crystallographic analysis. All isolated compounds (1-20) exhibited varying degrees of antiviral efficacy against respiratory syncytial virus (RSV). Notably, the anti-RSV activity of compound 12 (IC50 5.01 ± 0.14 μmol·L-1) surpassed that of the positive control (ribavirin, IC50 6.23 ± 0.95 μmol·L-1), as validated through plaque reduction and immunofluorescence assays. The identification of anti-RSV compounds from P. harmala seeds may enhance the development and application of this plant in antiviral therapeutic products.
Antiviral Agents/isolation & purification*
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Tryptamines/isolation & purification*
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Peganum/chemistry*
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Seeds/chemistry*
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Alkaloids/isolation & purification*
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Molecular Structure
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Humans
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Respiratory Syncytial Viruses/drug effects*
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Plant Extracts/pharmacology*
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Drugs, Chinese Herbal/pharmacology*
4.MYB polymorphism molecular marker: A novel molecular marker for authenticity and geographical origin identification of Citri Reticulatae Pericarpium.
Qiqing CHENG ; Ziyu TANG ; Yue OUYANG ; Chunsong CHENG ; Chichou LAO ; Hao CUI ; Hua ZHOU ; Yongshu LIANG
Chinese Herbal Medicines 2025;17(2):296-306
OBJECTIVE:
Citri Reticulatae Pericarpium (Chenpi, CRP) is one of the most used traditional Chinese medicines with great medicinal, dietary and collection values, among which the Citrus reticulata cv. 'Chachi' (Citrus reticulata cv. Chachiensis) from Guangdong Xinhui is the geoherb of CRP. Xinhui CRP in the market was often counterfeited with other varieties or origins, molecular identification method can effectively distinguish different CRP varieties, but it's still a difficult problem to identify the same CRP variety from different origin. It is necessary to discover a new molecular marker to ensure the safe and effective application of Xinhui CRP.
METHODS:
We selected one of the most studied transcription factor families in Citrus genus, MYB, to design the specific candidate primers on the conserved region. The primers with good band repeatability and high polymorphism were screened for PCR amplification of the test materials, and the genetic similarity coefficient among different families, genera, species, and origins were calculated. The cluster analysis was performed by unweighted pair group method using arithmetic average (UPGMA).
RESULTS:
A total of ten MYB primers were screened out to identify Xinhui CRP from plants from different family (Panax ginseng and Morus alba), genus (Clausena lansium and Zanthoxylum schinifolium), and species (Citrus reticulata, C. sinensis and C. maxima). Furthermore, two from the ten primers, M1 and M10, were found to distinguish Xinhui CRP from other origins. There were 169, 113, 133 and 134 polymorphic bands in the identification of different families, genera, species, and origins respectively, and the accordingly polymorphism ration were 79.88%, 76.87%, 79.20% and 82.84%. Moreover, M1 was discovered to be the best primer to identify Xinhui CRP from other seven origins, the cluster analysis results based on the genetic similarity coefficients were consistent with the geographical distribution.
CONCLUSION
This study established a novel molecular identification method according to MYB transcription factor, which can analyze the potential parental relationship of CRP germplasm, as well as identify the quality and origins of Xinhui CPR.
5.Predictive Modeling of Symptomatic Intracranial Hemorrhage Following Endovascular Thrombectomy: Insights From the Nationwide TREAT-AIS Registry
Jia-Hung CHEN ; I-Chang SU ; Yueh-Hsun LU ; Yi-Chen HSIEH ; Chih-Hao CHEN ; Chun-Jen LIN ; Yu-Wei CHEN ; Kuan-Hung LIN ; Pi-Shan SUNG ; Chih-Wei TANG ; Hai-Jui CHU ; Chuan-Hsiu FU ; Chao-Liang CHOU ; Cheng-Yu WEI ; Shang-Yih YAN ; Po-Lin CHEN ; Hsu-Ling YEH ; Sheng-Feng SUNG ; Hon-Man LIU ; Ching-Huang LIN ; Meng LEE ; Sung-Chun TANG ; I-Hui LEE ; Lung CHAN ; Li-Ming LIEN ; Hung-Yi CHIOU ; Jiunn-Tay LEE ; Jiann-Shing JENG ;
Journal of Stroke 2025;27(1):85-94
Background:
and Purpose Symptomatic intracranial hemorrhage (sICH) following endovascular thrombectomy (EVT) is a severe complication associated with adverse functional outcomes and increased mortality rates. Currently, a reliable predictive model for sICH risk after EVT is lacking.
Methods:
This study used data from patients aged ≥20 years who underwent EVT for anterior circulation stroke from the nationwide Taiwan Registry of Endovascular Thrombectomy for Acute Ischemic Stroke (TREAT-AIS). A predictive model including factors associated with an increased risk of sICH after EVT was developed to differentiate between patients with and without sICH. This model was compared existing predictive models using nationwide registry data to evaluate its relative performance.
Results:
Of the 2,507 identified patients, 158 developed sICH after EVT. Factors such as diastolic blood pressure, Alberta Stroke Program Early CT Score, platelet count, glucose level, collateral score, and successful reperfusion were associated with the risk of sICH after EVT. The TREAT-AIS score demonstrated acceptable predictive accuracy (area under the curve [AUC]=0.694), with higher scores being associated with an increased risk of sICH (odds ratio=2.01 per score increase, 95% confidence interval=1.64–2.45, P<0.001). The discriminatory capacity of the score was similar in patients with symptom onset beyond 6 hours (AUC=0.705). Compared to existing models, the TREAT-AIS score consistently exhibited superior predictive accuracy, although this difference was marginal.
Conclusions
The TREAT-AIS score outperformed existing models, and demonstrated an acceptable discriminatory capacity for distinguishing patients according to sICH risk levels. However, the differences between models were only marginal. Further research incorporating periprocedural and postprocedural factors is required to improve the predictive accuracy.
6.Predictive Modeling of Symptomatic Intracranial Hemorrhage Following Endovascular Thrombectomy: Insights From the Nationwide TREAT-AIS Registry
Jia-Hung CHEN ; I-Chang SU ; Yueh-Hsun LU ; Yi-Chen HSIEH ; Chih-Hao CHEN ; Chun-Jen LIN ; Yu-Wei CHEN ; Kuan-Hung LIN ; Pi-Shan SUNG ; Chih-Wei TANG ; Hai-Jui CHU ; Chuan-Hsiu FU ; Chao-Liang CHOU ; Cheng-Yu WEI ; Shang-Yih YAN ; Po-Lin CHEN ; Hsu-Ling YEH ; Sheng-Feng SUNG ; Hon-Man LIU ; Ching-Huang LIN ; Meng LEE ; Sung-Chun TANG ; I-Hui LEE ; Lung CHAN ; Li-Ming LIEN ; Hung-Yi CHIOU ; Jiunn-Tay LEE ; Jiann-Shing JENG ;
Journal of Stroke 2025;27(1):85-94
Background:
and Purpose Symptomatic intracranial hemorrhage (sICH) following endovascular thrombectomy (EVT) is a severe complication associated with adverse functional outcomes and increased mortality rates. Currently, a reliable predictive model for sICH risk after EVT is lacking.
Methods:
This study used data from patients aged ≥20 years who underwent EVT for anterior circulation stroke from the nationwide Taiwan Registry of Endovascular Thrombectomy for Acute Ischemic Stroke (TREAT-AIS). A predictive model including factors associated with an increased risk of sICH after EVT was developed to differentiate between patients with and without sICH. This model was compared existing predictive models using nationwide registry data to evaluate its relative performance.
Results:
Of the 2,507 identified patients, 158 developed sICH after EVT. Factors such as diastolic blood pressure, Alberta Stroke Program Early CT Score, platelet count, glucose level, collateral score, and successful reperfusion were associated with the risk of sICH after EVT. The TREAT-AIS score demonstrated acceptable predictive accuracy (area under the curve [AUC]=0.694), with higher scores being associated with an increased risk of sICH (odds ratio=2.01 per score increase, 95% confidence interval=1.64–2.45, P<0.001). The discriminatory capacity of the score was similar in patients with symptom onset beyond 6 hours (AUC=0.705). Compared to existing models, the TREAT-AIS score consistently exhibited superior predictive accuracy, although this difference was marginal.
Conclusions
The TREAT-AIS score outperformed existing models, and demonstrated an acceptable discriminatory capacity for distinguishing patients according to sICH risk levels. However, the differences between models were only marginal. Further research incorporating periprocedural and postprocedural factors is required to improve the predictive accuracy.
7.Predictive Modeling of Symptomatic Intracranial Hemorrhage Following Endovascular Thrombectomy: Insights From the Nationwide TREAT-AIS Registry
Jia-Hung CHEN ; I-Chang SU ; Yueh-Hsun LU ; Yi-Chen HSIEH ; Chih-Hao CHEN ; Chun-Jen LIN ; Yu-Wei CHEN ; Kuan-Hung LIN ; Pi-Shan SUNG ; Chih-Wei TANG ; Hai-Jui CHU ; Chuan-Hsiu FU ; Chao-Liang CHOU ; Cheng-Yu WEI ; Shang-Yih YAN ; Po-Lin CHEN ; Hsu-Ling YEH ; Sheng-Feng SUNG ; Hon-Man LIU ; Ching-Huang LIN ; Meng LEE ; Sung-Chun TANG ; I-Hui LEE ; Lung CHAN ; Li-Ming LIEN ; Hung-Yi CHIOU ; Jiunn-Tay LEE ; Jiann-Shing JENG ;
Journal of Stroke 2025;27(1):85-94
Background:
and Purpose Symptomatic intracranial hemorrhage (sICH) following endovascular thrombectomy (EVT) is a severe complication associated with adverse functional outcomes and increased mortality rates. Currently, a reliable predictive model for sICH risk after EVT is lacking.
Methods:
This study used data from patients aged ≥20 years who underwent EVT for anterior circulation stroke from the nationwide Taiwan Registry of Endovascular Thrombectomy for Acute Ischemic Stroke (TREAT-AIS). A predictive model including factors associated with an increased risk of sICH after EVT was developed to differentiate between patients with and without sICH. This model was compared existing predictive models using nationwide registry data to evaluate its relative performance.
Results:
Of the 2,507 identified patients, 158 developed sICH after EVT. Factors such as diastolic blood pressure, Alberta Stroke Program Early CT Score, platelet count, glucose level, collateral score, and successful reperfusion were associated with the risk of sICH after EVT. The TREAT-AIS score demonstrated acceptable predictive accuracy (area under the curve [AUC]=0.694), with higher scores being associated with an increased risk of sICH (odds ratio=2.01 per score increase, 95% confidence interval=1.64–2.45, P<0.001). The discriminatory capacity of the score was similar in patients with symptom onset beyond 6 hours (AUC=0.705). Compared to existing models, the TREAT-AIS score consistently exhibited superior predictive accuracy, although this difference was marginal.
Conclusions
The TREAT-AIS score outperformed existing models, and demonstrated an acceptable discriminatory capacity for distinguishing patients according to sICH risk levels. However, the differences between models were only marginal. Further research incorporating periprocedural and postprocedural factors is required to improve the predictive accuracy.
8.The impact of frailty on the prognosis of chronic heart failure patients treated with cardiac resynchronization therapy defibrillator implantation
Liang GAO ; Huasheng LYU ; Baopeng TANG ; Xianhui ZHOU ; Xinchun CHENG ; Youquan SHI ; Yaodong LI ; Jianghua ZHANG ; Qiang XING ; Tuerhong ZUKELA ; Yanmei LU
Chinese Journal of Internal Medicine 2025;64(1):45-51
Objective:The aim of this study was to assess the frailty status of patients with heart failure undergoing CRT-D and then explore the predictive value of frailty for all-cause mortality and heart failure-related readmissions in these patients.Methods:We retrospectively included 374 patients with chronic heart failure who underwent CRT-D treatment at the First Affiliated Hospital of Xinjiang Medical University between June 2020 and June 2024. Based on the Tilburg Debilitation Assessment Scale, 175 patients (46.8%) were classified as frail while 199 (53.2%) were classified as non-frail. The baseline data between the two groups was compared using Cox regression analysis and Kaplan-Meier curves were used for survival analysis. P-values of <0.05 indicated statistically significant differences. Results:A total of 374 patients aged 25-93 (68±11) years were enrolled in this study, 101 (27.0%) of which were female. Among these, 175 (46.8%) were categorized as frail, and 199 (53.2%) were classified as non-frail. Over a median follow-up time of 23 (5, 45) months, 35 (9.4%) patients experienced all-cause mortality, with 30 (17.1%) deaths occurring in the frail group and 5 (2.5%) in the non-frail group; meanwhile, readmission events due to heart failure occurred in a total of 174 (46.5%) patients, including 122 (70.1%) in the frail group, and 52 (29.9%) in the non-frail group. Cox analysis showed that frailty was a significant determinant of all-cause mortality ( HR=21.25, 95% CI 3.99-113.30, P<0.001) and readmission among heart failure patients receiving CRT-D ( HR=2.52, 95% CI 1.73-3.68, P<0.001). Log-rank tests showed that the survival rate of patients in the frail group was significantly lower than that of patients in the non-frail group ( HR=7.22, 95% CI 2.80-18.60, P<0.001) and the risk of readmission events due to heart failure was significantly higher among patients in the frail group than among those in the non-frail group ( HR=2.75, 95% CI 1.98-3.81, P<0.001). Conclusions:Frailty is an independent predictor of postoperative all-cause mortality and the occurrence of heart failure-related readmissions in patients with heart failure treated receiving CRT-D.
9.Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique (version 2025)
Sihao HE ; Junchao XING ; Tongwei CHU ; Zhengqi CHANG ; Xigao CHENG ; Fei DAI ; Xiaobing JIANG ; Jie HAO ; Jiang HU ; Jinghui HUANG ; Tianyong HOU ; Fei LUO ; Bo LIAO ; Changqing LI ; Lei LIU ; Guodong LIU ; Peng LIU ; Sheng LU ; Weishi LI ; Yang LIU ; Zhen LIU ; Wei MEI ; Peifu TANG ; Bing WANG ; Bing WANG ; Ce WANG ; Hongli WANG ; Liang WANG ; Shengru WANG ; Xiaobin WANG ; Yang WANG ; Yingfeng WANG ; Zheng WANG ; Jianzhong XU ; Guoyong YIN ; Haiyang YU ; Qiang YANG ; Zhaoming YE ; Bin ZHANG ; Chengmin ZHANG ; Jun ZOU ; Qiang ZHOU ; Min ZHAO ; Rui ZHOU ; Xiaojun ZHANG ; Yongfei ZHAO ; Zhongrong ZHANG ; Zehua ZHANG ; Yingze ZHANG
Chinese Journal of Trauma 2025;41(11):1035-1047
For middle-aged and elderly patients with conditions such as spinal fractures and degenerative spinal diseases, spinal internal fixation is a core surgical procedure for reconstructing spinal stability, heavily relying on the biomechanical stability provided by pedicle screw systems. Whereas, these patients are often complicated by osteoporosis that can significantly compromise the stability of the bone-pedicle screw interface, leading to a marked increase in pedicle screw loosening and surgical failure rates. The bone cement-augmented pedicle screw technique, which involves injecting bone cement into the vertebral body or screw trajectory to optimize the mechanical properties of the bone-pedicle screw composite, has been proven to significantly enhance fixation strength and effectively prevent screw-related failures, thereby reducing the incidence of internal fixation failure in high-risk populations undergoing spinal fusion. However, the widespread clinical application of this technique has faced challenges such as inaccurate clinical decision-making (indication and contraindication selection), non-standardized operative practices, and insufficient awareness of complication prevention, resulting in considerable variability in clinical outcomes and even severe complications. To address this, Prof. Luo Fei from First Affiliated Hospital of Army Medical University initiated the project and the Chinese Association Orthopaedic Surgeons organized relevant experts to develop the Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique ( version 2025), based on current evidence. The guidelines put forward 8 recommendations regarding the clinical value, scope of application, and operational standards of the technique, aiming to provide evidence-based medical support and technical standardization for clinical decision-making.
10.Establishment and performance evaluation of a second-tier screening method for neonatal congenital adrenal hyperplasia
Chengfang TANG ; Dan CHENG ; Minyi TAN ; Fang TANG ; Liang LIN ; Xuefang JIA ; Xiang JIANG ; Yuan WANG ; Yonglan HUANG
Chinese Journal of Perinatal Medicine 2025;28(12):1115-1121
Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.

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