1.Correlation of emotional and behavioral problems among preschool children with maternal parenting competence and family rearing environment
FANG Ling, QIAN Duoduo, CHEN Zongfang, WU Qiong, GUO Ningtian, SHEN Jing, ZHAI Jinxia
Chinese Journal of School Health 2026;47(4):522-526
Objective:
To investigate the emotional and behavioral problems among preschool children and the correlations with maternal parenting competence and family rearing environment, so as to provide a scientific basis and practical guidance for the physical and mental health development of preschool children.
Methods:
In June 2025, 660 preschool children aged 3-6 years old were selected from 10 kindergartens in Hefei, Anqing, and Tongling in Anhui Province by using a stratified cluster random sampling method. A questionnaire survey was conducted among all parents of preschool children using the Parent version of the Strengths and Difficulties Questionnaire, the Child Family Rearing Environment Scale, and the Chinese version of the Parenting Sense of Competence Scale. Pearson correlation analysis and multiple linear regression analysis were used to analyze the related factors of emotional and behavioral problems among preschool children. SPSS macro program Process 4.1 and Bootstrap method were used to test the mediating effect of family parenting environment between emotional and behavioral problems among preschool children and maternal parenting competence.
Results:
The detection rate of emotional and behavioral problems among preschool children was 20.15%. The total scores of family rearing environment and maternal parenting competence were negatively correlated with emotional and behavioral problems among preschool children ( r =-0.45,-0.79), and the total score of family rearing environment was positively correlated with the total score of maternal parenting competence ( r =0.43) (all P <0.01). Multiple linear regression showed that, after controlling whether being only child, parents educational level, registered residence location and other variables, family rearing environment, self efficacy and satisfaction were all negative predictors of emotional and behavioral problems among preschool children ( B =-0.07, -0.42, -0.42, all P <0.01). The mediation effect results showed that maternal parenting competence could positively predict the family rearing environment ( B =0.75), while maternal parenting competence and family rearing environment could both negatively predict emotional and behavioral problems among preschool children ( B =-0.49, -0.06 ) (all P <0.05). The Bootstrap sampling method test results showed that the effect value of the indirect effect of maternal parenting competence on emotional and behavioral problems through the family rearing environment was -0.04, and the effect proportion was 8.21 %.
Conclusion
Preschool children with a better family rearing environment and stronger maternal parenting competence are less likely to have emotional and behavioral problems, among which the family rearing environment has a mediating effect.
2.The systemic inflammatory response index as a risk factor for all-cause and cardiovascular mortality among individuals with coronary artery disease: evidence from the cohort study of NHANES 1999-2018.
Dao-Shen LIU ; Dan LIU ; Hai-Xu SONG ; Jing LI ; Miao-Han QIU ; Chao-Qun MA ; Xue-Fei MU ; Shang-Xun ZHOU ; Yi-Xuan DUAN ; Yu-Ying LI ; Yi LI ; Ya-Ling HAN
Journal of Geriatric Cardiology 2025;22(7):668-677
BACKGROUND:
The association of systemic inflammatory response index (SIRI) with prognosis of coronary artery disease (CAD) patients has never been investigated in a large sample with long-term follow-up. This study aimed to explore the association of SIRI with all-cause and cause-specific mortality in a nationally representative sample of CAD patients from United States.
METHODS:
A total of 3386 participants with CAD from the National Health and Nutrition Examination Survey (NHANES) 1999-2018 were included in this study. Cox proportional hazards model, restricted cubic spline (RCS), and receiver operating characteristic curve (ROC) were performed to investigate the association of SIRI with all-cause and cause-specific mortality. Piece-wise linear regression and sensitivity analyses were also performed.
RESULTS:
During a median follow-up of 7.7 years, 1454 all-cause mortality occurred. After adjusting for confounding factors, higher lnSIRI was significantly associated with higher risk of all-cause (HR = 1.16, 95% CI: 1.09-1.23) and CVD mortality (HR = 1.17, 95% CI: 1.05-1.30) but not cancer mortality (HR = 1.17, 95% CI: 0.99-1.38). The associations of SIRI with all-cause and CVD mortality were detected as J-shaped with threshold values of 1.05935 and 1.122946 for SIRI, respectively. ROC curves showed that lnSIRI had robust predictive effect both in short and long terms.
CONCLUSIONS
SIRI was independently associated with all-cause and CVD mortality, and the dose-response relationship was J-shaped. SIRI might serve as a valid predictor for all-cause and CVD mortality both in the short and long terms.
3.Bear Bile Powder Ameliorates LPS-Induced Acute Lung Injury by Inhibiting CD14 Pathway and Improving Intestinal Flora: Exploration of "Fei (Lung)-Dachang (Large Intestine) Interaction" Theory.
Long CHENG ; Hui-Ling TIAN ; Hong-Yuan LEI ; Ying-Zhou WANG ; Ma-Jing JIAO ; Yun-Hui LIANG ; Zhi-Zheng WU ; Xu-Kun DENG ; Yong-Shen REN
Chinese journal of integrative medicine 2025;31(9):821-829
OBJECTIVE:
To explore the effect of bear bile powder (BBP) on acute lung injury (ALI) and the underlying mechanism.
METHODS:
The chemical constituents of BBP were analyzed by ultra-high-pressure liquid chromatography-mass spectrometry (UPLC-MS). After 7 days of adaptive feeding, 50 mice were randomly divided into 5 groups by a random number table (n=10): normal control (NC), lipopolysaccharide (LPS), dexamethasone (Dex), low-, and high-dose BBP groups. The dosing cycle was 9 days. On the 12th and 14th days, 20 µL of Staphylococcus aureus solution (bacterial concentration of 1 × 10-7 CFU/mL) was given by nasal drip after 1 h of intragastric administration, and the mice in the NC group was given the same dose of phosphated buffered saline (PBS) solution. On the 16th day, after 1 h intragastric administration, 100 µL of LPS solution (1 mg/mL) was given by tracheal intubation, and the same dose of PBS solution was given to the NC group. Lung tissue was obtained to measure the myeloperoxidase (MPO) activity, the lung wet/dry weight ratio and expressions of CD14 and other related proteins. The lower lobe of the right lung was obtained for pathological examination. The concentrations of inflammatory cytokines including interleukin (IL)-6, tumour necrosis factor α (TNF-α ) and IL-1β in the bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay, and the number of neutrophils was counted. The colonic contents of the mice were analyzed by 16 sRNA technique and the contents of short-chain fatty acids (SCFAs) were measured by gas chromatograph-mass spectrometer (GC-MS).
RESULTS:
UPLC-MS revealed that the chemical components of BBP samples were mainly tauroursodeoxycholic acid and taurochenodeoxycholic acid sodium salt. BBP reduced the activity of MPO, concentrations of inflammatory cytokines, and inhibited the expression of CD14 protein, thus suppressing the activation of NF-κB pathway (P<0.05). The lung histopathological results indicated that BBP significantly reduced the degree of neutrophil infiltration, cell shedding, necrosis, and alveolar cavity depression. Moreover, BBP effectively regulated the composition of the intestinal microflora and increased the production of SCFAs, which contributed to its treatment effect (P<0.05).
CONCLUSIONS
BBP alleviates lung injury in ALI mouse through inhibiting activation of NF-κB pathway and decreasing expression of CD14 protein. BBP may promote recovery of ALI by improving the structure of intestinal flora and enhancing metabolic function of intestinal flora.
Animals
;
Acute Lung Injury/pathology*
;
Lipopolysaccharides
;
Ursidae
;
Gastrointestinal Microbiome/drug effects*
;
Bile/chemistry*
;
Lipopolysaccharide Receptors/metabolism*
;
Powders
;
Male
;
Lung/drug effects*
;
Mice
;
Peroxidase/metabolism*
;
Signal Transduction/drug effects*
;
Cytokines/metabolism*
4.Prodrug-based combinational nanomedicine remodels lipid metabolism for reinforced ferroptosis and immune activation.
Ling LIN ; Zaixiang FANG ; Guohao LIU ; Yiwei LIU ; Zhiqian LI ; Dayi PAN ; Yunkun LI ; Hemi KANG ; Xiaoding SHEN ; Jingyao ZHANG ; Qiyong GONG ; Kui LUO ; Jing JING
Acta Pharmaceutica Sinica B 2025;15(5):2746-2763
Ferroptosis is a form of programmed cell death characterized by overwhelmed lipid oxidation, and it has emerged as a promising strategy for cancer therapy. Enhanced ferroptosis could overcome the limitations of conventional therapeutic modalities, particularly in difficult-to-treat tumors. In this study, we developed a dual-modality therapy in nanomedicine by combining paclitaxel (PTX) chemotherapy and pyropheophorbide-a (Ppa) phototherapy. Heparin (HP) was grafted with poly(N-(2'-hydroxy) propyl methacrylamide) (pHPMA) using reversible addition-fragmentation chain transfer polymerization to form HP-pHPMA (HH), which was utilized to deliver Ppa and PTX, yielding HP-pHPMA-Ppa (HH-Ppa) and HP-pHPMA-PTX (HH-PTX), respectively. The prodrug-based combinational nanomedicine (HH-PP) was formed by co-assembly of HH-PTX and HH-Ppa. It was found that HH-PP treatment significantly disrupted lipid metabolism in triple-negative breast cancer (TNBC) cells, induced extensive lipid oxidation, and promoted ferroptosis. In vivo, HH-PP intervention achieved a tumor growth inhibition rate of 86.63% and activated adaptive immunity with an elevated CD8+ cytotoxic T cell infiltration level. This combinational nanomedicine offers a promising platform for co-delivery of multiple therapeutic agents. It exerts a promising anti-tumor effect via enhanced ferroptosis and ferroptosis-induced immune activation by disrupting lipid metabolism in TNBC cancer cells.
5.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
6.Prospective cohort study of CEUS quantitative parameters combined with immunohistochemistry in predicting NAC curative efficacy for breast cancer
Qiqi SHEN ; Wenjuan WU ; Jing HUO ; Ling CHEN
China Medical Equipment 2025;22(9):56-61
Objective:To explore application value of a constructed predictive model of quantitative parameters of contrast-enhanced ultrasound(CEUS)combined with immunohistochemical indicators in assessing the curative efficacy of neoadjuvant chemotherapy(NAC)for breast cancer,so as to provide objective basis for clinically individual treatment decisions.Methods:The CEUS quantitative parameters were used to combine with immunohistochemical indicators to construct predictive model,and this study adopted prospective cohort design.A total of 93 patients who were preliminarily diagnosed as breast cancer at Affiliated Hospital of Jiangnan University during June 2022 and June 2023 were included in this study.According to the pathologically relieve condition,they were divided into significant response group(41 cases)and non-significant response group(52 cases).All of them received NAC with docetaxel/doxorubicin/cyclophosphamide(TAC).Before treatment,the peak intensity(PI),time-to-peak(TTP),and wash-in rate(WIR)of them were obtained through CEUS,and all of patients underwent immunohistochemical examination to detect the expressions of immunohistochemical indicators included estrogen receptor(ER),progesterone receptor(PR),human epidermal growth factor receptor 2(HER2)and Ki-67 proliferation index.The Miller-Payne grade G4-G5 was used as the standard of pathological complete response(pCR).Multivariate logistic regression was adopted to screen independent predictors,and construct a jointly predictive model,and verify effectiveness by Bootstrap resampling method.Results:The PI value of significant response group was(22.7±4.1)dB,which was significantly higher than(18.3±3.6)dB of non-significant response group,and the difference was significant(t=5.437,P<0.001).The TTP of significant response group was(14.2±2.8)s,which was shorter than(18.6±3.1)s of non-significant response group,and the difference was significant(t=7.152,P<0.05).The wash-in rate(WIR)of significant response group was(1.61±0.43)dB/s,which was significantly higher than(0.98±0.37)dB/s of non-significant response group,and the difference was significant(t=7.893,P<0.001).In the immunohistochemical indicators,the positive HER2 and high Ki-67 expression significantly correlated with pathological response.In the results of positive HER2,there were 17 cases(41.5%)in 41 patients of significant response group,and there were 9 cases(17.3%)in non-significant response group,and the positive HER2 of significant response group was higher than that of non-significant response group,and the difference was significant(x2=7.326,P<0.05).For patients whose Ki-67 were larger or equal to 20%,the positive rate of significant response group was 75.6%(31 cases),which was higher than 57.7%(30 cases)of non-significant response group.For patients whose Ki-67 were less than 20%,the positive rate of significant response group was 24.4%(10 cases),which was significantly higher than 42.3%(22 cases)of non-significant response group,and the difference was significant(x2=3.921,P<0.05).Multivariate analysis indicated that TTP≤15 s,WIR≥1.5 dB/s,and positive HER2 were respectively independent predictors(OR=4.23,3.76,2.91,P<0.05).The area under curve(AUC)value of receiver operating characteristic(ROC)curve of joint model was 0.89(95%CI:0.83-0.95),and the sensitivity and specificity of that were respectively 92.7%and 80.8%,which were significantly better than each single parameter.Decision curve analysis indicated that the net benefit value of joint model increased by 21.3%-28.6%than conventional strategy when threshold probabilities was 15%-60%.Conclusion:CEUS quantitative parameters(TTP,WIR)that combine with HER2 status can construct predictive model with high-precision and low-cost for NAC curative efficacy,which synergistic effect in dynamic perfusion assessment and molecularly pathological characteristic can provide new paradigm for precision treatment in breast cancer.This mode has excellent clinical applicability,and can effectively identify chemosensitive populations and optimize decision-making process of treatment.
7.Early- and mid-term outcomes of using porous-coated metaphyseal sleeves to reconstruct severe bone defects in revision total knee arthroplasty
Penghua MA ; Tingxian LING ; Fuxing PEI ; Jing YANG ; Pengde KANG ; Bin SHEN ; Zongke ZHOU
Chinese Journal of Orthopaedic Trauma 2025;27(6):479-484
Objective:To explore the early- to mid-term therapeutic efficacy of using porous-coated metaphyseal sleeves to reconstruct severe bone defects in revision total knee arthroplasty (rTKA).Methods:A retrospective analysis was conducted of the clinical data of the 39 patients (40 knees) who had undergone rTKA by porous-coated metaphyseal sleeve reconstruction at Department of Orthopaedics, West China Hospital, Sichuan University between May 2017 and September 2023. The cohort included 6 males (6 knees) and 33 females (34 knees), with an age of (67.0±9.7) years. The revision was to cure periprosthetic infection after TKA in 12 knees, to correct prosthesis loosening in 19 knees, to treat periprosthetic fracture in 4 knees, to stabilize postoperative joint instability in 4 knees, and to manage postoperative joint stiffness in 1 knee. All patients underwent standard revision procedures, including removal of the original prosthesis, management of bone defects, implantation of revision prosthesis, and adjustment of ligamentous balance and fixation. The patients' surgical time, intraoperative blood loss, incidence of complications, as well as visual analogue scale (VAS), knee range of motion, and Hospital for Special Surgery (HSS) knee joint scores at the last follow-up were recorded.Results:The surgical time was (2.7±0.8) hours, and intraoperative blood loss (337.5±165.4) mL for this cohort. All the 39 patients were followed up for (4.8±2.1) years after surgery. At the last follow-up, their VAS pain score was 2.0 (1.0, 2.0) points, their knee range of motion reached 116.3°±12.2°, and their total score, pain score, and function score of the HSS system were respectively 87.0 (82.8, 89.3) points, 25.0 (22.8, 29.0) points, and 61.0 (60.0, 62.0) points, all showing statistically significant improvements compared with their preoperative values [(6.8±1.7) points, 70.4°±15.2°, (43.1±9.6) points, (9.3±3.1) points, and (33.8±10.1) points] ( P<0.05). In all patients, incisions healed at one stage after surgery, and no complications such as deep vein thrombosis or neurovascular injury occurred. Complications included popliteal artery thrombosis in 1 patient (1 knee) immediately after surgery, acute infection in 1 patient (1 knee) at 3 years after surgery, and periprosthetic fracture due to a traffic accident in 1 patient (1 knee) at 4 years after surgery, and distal prosthesis-related pain in 3 patients (3 knees). Conclusion:Use of porous-coated metaphyseal sleeves in rTKA to reconstruct severe bone defects exhibits favorable early- to mid-term therapeutic outcomes.
8.A Rubric System for Evaluating the TCA-based Ideological and Political Teaching Model:Its Construction and Application
Ying WANG ; Ling-Hui LV ; Ren-Ji WEI ; Xiang ZHANG ; Yi RU ; Bo YAN ; Lan SHEN ; Mao SUN ; Liang LIANG ; Jing ZHAO
Chinese Journal of Biochemistry and Molecular Biology 2025;41(1):53-67
The success of"New Medical Sciences"in higher education requires effective tools in evalua-ting students'performance in courses.Previously,we reported a teamwork(T),critique(C)and ap-preciation(A)(TCA)ideological and political model,a teaching model widely applied in Basic Medical courses.TCA is an abbreviation derived from Tricarboxylic Acid Cycle in Biochemistry as an analogy for nurturing the abilities of thinking and teamwork(T),critique(C)and appreciation(A),which hope-fully could provide students with moral norms for cognition,science and life.This paper further explores the tools to assess the educational outcomes of the TCA model,by which teachers can collect feedback and reflect on teaching quality and effectiveness.Addressing the challenges of individual differences in large classes,fragmented learning feedback,and the difficulty of measuring meta-cognition in educational evaluation,this study employs strategies of value-added assessment,matrix assessment and norm-transfer-able assessment to evaluate the TCA abilities from the aspects of thinking quality,thinking creativity,co-operation ability,iterative thinking,dialectical thinking and job responsibility.By modifying/using 18 e-valuation tools in Education and Psychology,we have established a rubric system composed of 30 primary indicators(with 11 newly designed,10 partly modified and 9 directly adopted),along with 49 secondary indicators and 98 tertiary indicators to enhance the feasibility of the evaluation process.This rubric sys-tem was applied to Biochemistry teaching among the five-year-program undergraduates at Air Force Medi-cal University.Specifically,thinking and teamwork are evaluated by creative works from"the magic bio-chemical-circle",while critiques are assessed in large classes under the guidance of basic and clinical teachers,coupled with appreciation measured by job responsibility in a task-driven virtual reality(VR)project.The results indicate that Biochemistry teaching not only accumulates knowledge in students,but also achieves the goals in nurturing values and cognition.The inclusion of creative performance evalua-tion,cooperative learning and clinical case studies,can enhance students'interpersonal skills,coopera-tion,quality of thinking,creative thinking,iterative thinking and dialectical thinking to varying degrees.TCA-based Biochemistry teaching has a long-lasting impact on character education,and is capable of in-ducing positive long-term changes in students'cognition and lifestyle.Taken together,with the help of this rubric system,teachers can promptly acknowledge the effectiveness of their teaching,thereby facilita-ting their teaching strategies.
9.Experimental study on alternative method of local lymph node assay using bromodeoxyuridine with flow cytometry(LLNA:BrdU-FCM)for skin sensitization evaluation of cosmetics
Xiao-jun LYU ; Ju ZHANG ; Sen WU ; Xiao-ling XU ; Meng-ting SHI ; Jin-jing XU ; Wang-ping PAN ; Jia-te SHEN ; Kai-yong HE
Chinese Pharmacological Bulletin 2025;41(4):793-799
Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25%hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25%HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90%,specificity of 100%,positive prediction rate of 100%,negative prediction rate of 83%,false positive rate of 0%,false negative rate of 17%and accuracy of 93%.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.
10.Experimental study on alternative method of local lymph node assay using bromodeoxyuridine with flow cytometry(LLNA:BrdU-FCM)for skin sensitization evaluation of cosmetics
Xiao-jun LYU ; Ju ZHANG ; Sen WU ; Xiao-ling XU ; Meng-ting SHI ; Jin-jing XU ; Wang-ping PAN ; Jia-te SHEN ; Kai-yong HE
Chinese Pharmacological Bulletin 2025;41(4):793-799
Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25%hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25%HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90%,specificity of 100%,positive prediction rate of 100%,negative prediction rate of 83%,false positive rate of 0%,false negative rate of 17%and accuracy of 93%.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.


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