1.Effect of Zuogui Wan and Yougui Wan on Mitochondrial Biogenesis in BMSCs Through PGC-1α/PPARγ
Ying YANG ; Xiuzhi FENG ; Yiran CHEN ; Zhimin WANG ; Xian GUO ; Yanling REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):28-36
ObjectiveBased on the TCM theory of "Yang transforms materials to Qi while Yin constitutes material form", this paper explored the effects of Zuogui Wan and Yougui Wan on the molecular mechanism of mitochondrial biogenesis during the adipogenic differentiation process of rat bone marrow mesenchymal stem cells (BMSCs) by mediating peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and peroxisome proliferators-activated receptor γ (PPARγ), providing theoretical support for the prevention and treatment of postmenopausal osteoporosis (PMOP) using Zuogui Wan and Yougui Wan. MethodsBMSCs were divided into a blank group, Zuogui Wan (ZGW) group, Yougui Wan (YGW) group, and Progynova group. Cell identification was performed using flow cytometry. The growth curves of BMSCs were plotted using the methylthiazolyldiphenyl-tetrazolium bromide (MTT) method, and the effects of Zuogui Wan and Yougui Wan on the proliferation of BMSCs were detected. The Oil red O staining method was used to detect lipid droplet formation. The Western blot method was used to detect the expression of adipogenesis-related factors PPARγ, CCAAT/enharcer-binding protein (C/EBP)α, C/EBPβ, lipoprotein lipase (LPL) protein, brown adipose tissue-related (BAT) proteins PGC-1α, uncoupcing protein 1 (UCP1), PR domdin-containing protein 16 (PRDM16), mitochondrial biogenesis-related PGC-1α, nuclear respiratory factor 1 (Nrf1), nuclear factor E2-related factor 2 (Nrf2), and mitochondrial transcription factor A (TFAM). The expression of adipogenesis-related factors PPARγ, C/EBPα, C/EBPβ, LPL genes, and the copy number of cytochrome B (CytoB mtDNA) gene was detected using real-time polymerase chain reaction (Real-time PCR). Mitochondrial ultrastructure was detected using transmission electron microscopy. ResultsCompared with that in the blank group, the proliferation ability of BMSCs in each treatment group increased continuously as the intervention progressed, and lipid droplets significantly decreased after the drug intervention. The mRNA and protein expression levels of adipogenesis-related factors PPARγ, C/EBPα, C/EBPβ, and LPL were significantly downregulated (P<0.01), while those of the BAT-related factors PGC-1α, UCP1, PRDM16 were significantly upregulated (P<0.01). The number of mitochondria increased, accompanied by reduced swelling. The double membrane and cristae structure were clear, and the internal cristae rupture was reduced. The copy number of CytoB mtDNA in each treatment group was significantly increased (P<0.01). The protein expression levels of mitochondrial biogenesis-related PGC-1α, Nrf1, Nrf2, and TFAM in each treatment group were significantly increased (P<0.01). ConclusionBoth Zuogui Wan and Yougui Wan can prevent and treat PMOP by intervening in mitochondrial biogenesis in BMSCs through PGC-1α/PPARγ.
2.Network structure characteristics of trait aggression in children and adolescents based on psychometric network analysis
WANG Xu, LIU Yanling, WEI Mingchen, ZHU Ni, GENG Yibo, LIU Weijun, CHEN Shuai
Chinese Journal of School Health 2025;46(7):975-979
Objective:
To explore the core features of trait aggression in children and adolescents, so as to provide a theoretical basis for behavioral interventions targeting the central psychological characteristics of aggression in children and adolescents.
Methods:
From March to May 2020, a simple random convenience sampling method was employed to recruit 39 165 students from grades 4 to 12 in Sichuan, Chongqing, Guizhou, and Shandong. Data were collected via online questionnaires, with all participants completing the Chinese Version of the Aggression Questionnaire. Psychometric network analysis was utilized for data processing.
Results:
Trait aggression among Chinese children and adolescents was at a moderately low level. The core nodes of the network structure included physical aggression [if someone intentionally causes trouble for me, I will hit them severely (AGG6); if someone hits me, I will retaliate (AGG11)] and self aggression [When I am very irritable, I think of hurting myself (AGG5); when I am in a bad mood, I engage in behaviors that harm my health, such as overeating (AGG25)]. Across grade levels, core nodes primarily originated from the anger dimension [When I m angry, I feel like a powder magazine that could explode at any moment (AGG13); I can t control my temper (AGG18); I am prone to getting angry when I see things that are not pleasing to the eye (AGG23); I will get angry for no reason (AGG27)]. Except for grades 7 and 9, core nodes in other grades included the verbal aggression dimension [I am prone to arguments with people (AGG22)]. Before grade 8, core nodes incorporated the self aggression dimension (AGG 5, AGG 25); after grade 8, core nodes included the physical aggression dimension [AGG 6, AGG 11, I fight slightly more than others (AGG16), and if people around me make things difficult for me to a certain extent, I will fight with them (AGG26)]. No statistically significant differences were found in the trait aggression network structures across grades, genders, or within gender comparisons of different grades.
Conclusion
These findings broaden our understanding of aggression in children and adolescents, suggesting that behavioral interventions can effectively reduce aggressive behaviors in this population.
3.Mechanism of Modified Erxian Decoction Regulating Perimenopausal Syndrome via SIRT1/Kisspeptin/GnRH Signaling Pathway
Ruiyu HUANG ; Fang LEI ; Wuchaonan LIU ; Jingjing YANG ; Qianru ZENG ; Shengping LUO ; Yanling CHEN ; Mengge ZHANG ; Fanshun SHEN ; Yihui DENG ; Dingxiang LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):51-62
ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal (HPG) axis by modified Erxian decoction in rats with perimenopausal syndrome (PMS) and to further analyze the expression of proteins related to the silent information regulator 1 (SIRT1)/hypothalamic kisspeptin (Kisspeptin)/gonadotropin-releasing hormone (GnRH) signaling pathway in the arcuate nucleus region (ARC) of the hypothalamus, so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method, with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling, including a sham operation group, a model group, an estradiol valerate group (0.09 mg∙kg-1∙d-1), high-, medium-, and low-dose modified Erxian decoction groups (7.614, 3.807,1.903 5 g∙kg-1∙d-1), with 8 rats in each group. The estradiol valerate group and the high-, medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days, and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH). Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to determine the expression levels of SIRT1, Kisspeptin, kisspeptin receptor (GPR54), and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor (GnRH-R) in pituitary. ResultsCompared with the sham operation group, rats in the model group had a significantly increased body weight (P0.01), reduced wet weight and index of uterus (P0.01), endometrial thinning or atrophy, glandular atrophy, and a decreasing number of glands. Additionally, serum levels of E2 and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased (P0.01). Serum levels of FSH, LH, and GnRH, the expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus, and GnRH-R in pituitary significantly increased (P0.01). Compared with the model group, the estradiol valerate group and the high-, medium-dose modified Erxian decoction groups had significantly reduced body weight, serum levels of FSH, LH, and GnRH, and expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary (P0.05, P0.01) and significantly increased wet weight and index of uterus, serum level of E2, and expression of SIRT1 in the ARC region of the hypothalamus (P0.05, P0.01). In addition, they showed thickened endometrium, increased number of endometrial glands, and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets, and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus, the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway, the regulation of the expression of GnRH-R in the pituitary, the restoration of secretion balance of gonadotropins, and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.
4.Establishment and application of drug use evaluation criteria for aspirin enteric-coated tablets
Meimei LIN ; Jing ZHANG ; Caiyun ZHENG ; Yan CHEN ; Yanling GAO ; Shicai CHEN ; Jinhua ZHANG
Chinese Journal of Pharmacoepidemiology 2024;33(5):489-499
Objective To establish the drug use evaluation(DUE)criteria for aspirin enteric-coated tablets and provide a reference for the rationally clinical application of aspirin enteric-coated tablets.Methods DUE criteria for aspirin enteric-coated tablets were established from three aspects of indications,medications and medication results with reference to drug instructions of aspirin enteric-coated tablets,related guidelines,expert consensus and literature,and through Delphi method.A retrospective analysis was conducted to evaluate the medical records of patients who took aspirin enteric-coated tablets from January 2021 to June 2022 in Fuqing Hospital affiliated to Fujian Medical University.Results A total of 1 071 medical records were included.683 cases fully met the DUE criteria,with a rational rate of 63.77%.Irrational drug use mainly included inappropriate indications(6.26%),off-label drug use without approval(28.48%),contraindications(1.03%),inappropriate usage and dosage(1.68%),drug interactions with potential clinical significance(0.65%)and other inappropriate drug use(2.71%).Conclusion The established DUE standard for aspirin enteric-coated tablets has strong scientific practicability and feasibility.The irrational rate of aspirin enteric-coated tablets in this hospital is high.Corresponding intervention measures should be formulated to ensure the safety of clinical medication.
5.The development and implementation of a 3D technology-based female bed urinal
Yanling CHEN ; Hongyan LI ; Xiaobo WANG ; Shanshan XU ; Yuhong YAO ; Ping WANG ; Xiaomei SUN
Chinese Journal of Nursing 2024;59(18):2297-2300
Objective To utilize a 3D technology in the design of a female bed urinal and to evaluate its clinical efficacy.Methods A total of 102 adult female fracture patients with normal urination function admitted to a tertiary hospital in Hangzhou City from October 2022 to June 2023 were included in the study.They were divided into a control group(n=51)and an experimental group(n=51)according to random number method.Patients in control group used a regular urinal,while patients in the experimental group used the 3D technology-based female bed urinal.The level of physical pain caused by urination,the rate of urine immersion in the sacrococcygeal or gluteal cleft and the rate of bed unit or clothing of contamination were compared between the 2 groups.Results There was no significant difference in the rate of bed unit or clothing contamination between the 2 groups(P>0.05).However,the experimental group experienced significantly lower pain caused by urination,a lower rate of urine impregnation in the sacrococcygeal or gluteal fissure(P<0.001),compared to the control group.Conclusion The 3D technology-based female bed urinal has reasonable structure and simple operation,which can significantly reduce the physical pain caused by the change of body position,reduce the incidence of urine immersion events.
6.Ultrasound shear wave elastography of skin in diagnosis of lymphedema of lower extremity: a preliminary study
Jiaping LI ; Jia LUO ; Manying LI ; Jian QI ; Xiang ZHOU ; Qiushuang LI ; Shaozhen CHEN ; Xiaoyan XIE ; Yanling ZHENG
Chinese Journal of Microsurgery 2024;47(4):382-387
Objective:To explore a better measurement mode of shear wave elastography (SWE) in the skin of lymphedema limbs, and to test its diagnostic efficacy in lymphedema.Methods:Between 1st and 10th August, 2023, 22 healthy volunteers were recruited in the Department of Medical Ultrasonics, Institute of Diagnostic and Interventional Ulrasound, the First Afiliaed Hospital of Sun Yat-Sen University to measure the Young's modulus (E-value) of limb skin by SWE using a gel pad (GP group) and the thick-layer coupling gel (CG group) respectively. Then between 15th August and 28th September, 2023, 11 patients with 13 lower limb lymphedema, who were treated in the Department of Microsurgery, Orhopaedic Trauma and Hand Surgery, the First Affiliaed Hospital of Sun Yat-Sen University, were consecutively enrolled to find out the E-value of skin in oedematous limbs. The receiver operating characteristic curve (ROC) was constructed and the sensitivity, specificity, positive predictive value, negative predictive value and accuracy were calculated to evaluate the diagnostic efficiency. SPSS 26.0, R studio and GraphPad Prism 8 software were used for statistical analysis. Paired sample non-parametric test (Wilcoxon signed rank test) was used to compare the difference in E-value between the 2 groups. P<0.01 was considered statistically significant. Spearman correlation test was used to analyse the correlation of E-value values between the GP group and CG group. Results:Overall, it was found from the healthy volunteers that E-value of skin in distal limbs were higher than that in proximal limbs, especially in lower extremities. E-value of all scanned location in GP group were found higher than those of CG group with statistically significant difference ( P<0.01), and the values of interclass correlation coefficient (ICC) showed a good repeatability. The 11 patients with lymphedema in 13 lower extremities were examined with the thick-layer coupling gel. The skin E-value in calf of patients with lymphedema were significantly higher than that of the healthy volunteers ( P<0.01), except the skin of thighs ( P>0.01). A lymphedema was diagnosed while either a skin E-value was 27.6 kPa calculated by Youden index or with the thickness of skin was 2.3 mm. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy in the diagnosis of a lymphedema were found at 92.3%, 100%, 100%, 95.7% and 97.1%, respectively. The area under curve (AUC) of ROC was 0.962. The correlation coefficients (R) of E-value in arm, forearm, thigh and lower leg in the GP group and CP group were 0.665, 0.882, 0.850 and 0.815, respectively, which were all significantly correlated. Conclusion:Application of thick layer coupler in ultrasound skin SWE is highly feasible, and the combination of ultrasound skin SWE and skin thickness has higher diagnostic efficiency in the diagnosis of lower extremities lymphedema. It allows more accurate ultrasonic image technical support for early monitoring and diagnosis of lymphedema, microsurgical treatment and a quantitatively perioperative evaluation.
7.Differences in estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2 and Ki-67 expressions between primary sites and metastatic sites in metastatic triple-negative breast cancer
Na ZHOU ; Yanling HE ; Qian WANG ; Congying YANG ; Hao CHEN
Cancer Research and Clinic 2024;36(8):583-589
Objective:To investigate the differences in estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 expressions between metastatic sites and primary sites in metastatic triple-negative breast cancer (TNBC) patients and to analyze their effects on the prognosis of patients.Methods:A retrospective cohort study was conducted. The clinicopathological data of 108 patients diagnosed with metastatic TNBC in Lianyungang Hospital Affiliated to Xuzhou Medical University from September 2018 to September 2023 were collected. Local metastatic sites included ipsilateral axillary lymph nodes and chest wall, while distant metastatic sites encompassed contralateral axillary lymph nodes and chest wall, bilateral supraclavicular lymph nodes, cervical lymph nodes, bone and viscera. The metastatic sites were identified as metastatic TNBC by using immunohistochemistry (IHC) and/or fluorescence in situ hybridization. The heterogeneity in ER, PR, HER2, and Ki-67 expression between primary sites and metastatic sites in all patients and those with local and distant metastases was analyzed, and the heterogeneity was defined as the inconsistency in the expression of markers including loss and gain expressions between primary sites and metastatic sites. The Kaplan-Meier method was used to analyze disease-free survival (DFS), and log-rank test was used for comparison among groups.Results:All patients were female, with an average age of (55±10) years; 51 cases had local metastases, and 57 cases had distant metastases. In metastatic sites, no ER and PR positivity were observed; in primary sites, 28.7% (31/108) of patients were ER positive and 21.3% (23/108) of patients were PR positive, and there were statistically significant differences in the proportion of patients with ER or PR positive between primary sites and metastatic sites (all P < 0.001). There were no statistically significant differences in the proportions of patients with different expression status including HER2 [positive: 0 (0/108) vs. 4.6%(5/108), low expression: 50.0% (54/108) vs. 45.4% (49/108), negative: 50.0% (54/108) vs. 50.0% (54/108)], Ki-67 [high expression: 86.1% (93/108) vs. 91.7% (99/108)] between metastatic sites and primary sites (all P > 0.05). The proportions of patients with inconsistent ER and PR expression between metastatic sites and primary sites were 28.7% (31/108) and 21.3% (23/108), respectively, all due to the expression loss in metastatic sites; the proportions of patients with inconsistent HER2 and Ki-67 expression between metastatic sites and primary sites were 42.6% (46/108) and 11.1% (12/108), respectively, with HER2 and Ki-67 expression loss of metastatic sites accounting for 22.2% (24/108) and 8.3% (9/108), respectively and expression gain accounting for 20.4% (22/108) and 2.8% (3/108), respectively. The proportions of patients with inconsistent ER [45.6% (26/57) vs. 9.8% (5/51)], PR [36.8% (21/57) vs. 3.9% (2/51)] and Ki-67 [17.5% (10/57) vs. 3.9% (2/51)] expression in distant metastatic sites and primary sites were higher than those with local metastatic sites and primary sites, and the differences were statistically significant (all P < 0.05). The proportions of patients with inconsistent HER2 between distant metastatic sites and primary sites and those with inconsistent HER2 between local metastatic sites and primary sites were 47.4% (27/57), 37.3% (19/51), respectively, and the difference was not statistically significant ( P = 0.300). Patients with inconsistent ER and Ki-67 expressions had better DFS than those with consistent expressions, with median DFS time of 30 months (95% CI: 22-40 months) vs. 22 months (95% CI: 22-24 months) for ER and 28 months (95% CI: 20-61 months) vs. 22 months (95% CI: 22-24 months) for Ki-67, and the differences were statistically significant (all P < 0.05). There were no significant differences in DFS between metastatic sites and primary sites with consistent expressions of PR and HER2 or not (all P > 0.05). Conclusions:There are differences in ER and PR expressions between primary sites and metastatic sites of metastatic TNBC patients, while the expressions of HER2 and Ki-67 seem to be no differences. The inconsistency of ER, PR and Ki-67 expressions with primary sites in distant metastatic sites are more common compared with in local metastatic sites. The differences in hormone receptor expression between primary sites and metastatic sites may impact patients' DFS.
8.Therapeutic Effects of Type Ⅰ Tympanoplasty by the Continuous Irrigating Endoscopic Ear Surgery Mode
Chaoqiu XIAO ; Wei LI ; Hao WANG ; Yanling ZHANG ; Rui DING ; Bai CHEN ; Zeqi ZHAO
Journal of Audiology and Speech Pathology 2024;32(6):528-531
Objective To study the therapeutic effect of the type Ⅰ tympanoplasty by the continuous irriga-ting endoscopic ear surgery mode.Methods A total of 50 patients with chronic suppurative otitis media,who pre-pared for the type Ⅰ tympanoplasty in our department,were divided into the traditional operation mode group(20 cases)and the continuous irrigating mode group(30 cases).All patients underwent underlay myringoplasty with tragus cartilage and perichondria limplant.The operation time and postoperative outcome of the two groups were compared and analyzed.Results The mean operation time of the traditional group and the irrigating operation group was 66.10±2.43 minutes and 46.81±5.12 minutes respectively,and there was statistical difference between the two groups(P<0.05).The tympanic membrane healing rate of the traditional group and the irrigating operation group was 100.00%and 96.67%respectively,and there was no statistical difference between the two groups(P>0.05).The mean air conduction threshold and the value of air-bone gap were significantly reduced at 2 months after operation compared with that before operation(P<0.05).There was no significant difference between the two groups in the average pre and post-operative air-bone gaps(P>0.05).The incidence rates of postoperative compli-cations were 10.00%and 3.33%,respectively,with no statistical significance(P>0.05).Conclusion The contin-uous irrigating endoscopic ear surgery mode is safe and effective.Under the premise of ensuring clinical efficacy,the operation time is shortened,and it is worth popularizing in middle ear surgery.
9.Long-term trend of the prevalence of Kashin-Beck disease in Gansu Province
Shaolun YANG ; Xiaoyan CHEN ; Xin ZHENG ; Faqing CHEN ; Yanling WANG ; Xiulan FEI ; Xiaoning LIU
Chinese Journal of Endemiology 2024;43(5):383-387
Objective:To analyze the monitoring data of Kashin-Beck disease (KBD) in Gansu Province and learn about the long-term trend of KBD incidence.Methods:The X-ray detection rate of children with KBD in Gansu Province from 1990 to 2021 was collected through Gansu Provincial Center for Disease Control and Prevention. Joinpoint regression model was used to analyze the trend of X-ray detection rate of KBD in children. The annual percent change (APC) and average annual percent change (AAPC) were used to analyze the trend of the observed indicators. In addition, a polynomial regression model was constructed to fit the change of X-ray detection rate of children with KBD over time.Results:From 1990 to 2021, a total of 126 726 children were examined for KBD by X-ray in Gansu Province, with 3 011 positive cases, X-ray detection rate of KBD in children was 0 in 2019 - 2021. Joinpoint regression model analysis showed that the X-ray detection rate of children with KBD in Gansu Province from 1990 to 2018 showed a decreasing trend (AAPC = - 14.40%, P = 0.001). The APC of 1990 - 1998, 2002 - 2008 and 2008 - 2012 was - 17.75%, - 21.89% and - 49.89%, respectively, showing a decreasing trend ( P < 0.05). The APC of 2012 - 2018 was 6.15%, but the trend change was not statistically significant ( P = 0.475). The curve fitting of X-ray detection rate of KBD in children in Gansu Province over time was carried out, and the quadric polynomial equation was y = 0.000 314 5 x4 - 0.021 37 x3 + 0.487 1 x2 - 4.635 x + 22.08. Conclusions:The condition of KBD in children of Gansu Province has been effectively controlled. The X-ray detection rate has remained at a relatively low level since 2012, and it has reached a state of elimination since 2019.
10.Tracking and evaluation of the application of the standard of "Delimitation and Classification of Keshan Disease Areas" (GB 17020-2010)
Faqing CHEN ; Ping LI ; Suqin YU ; Yanling WANG ; Li ZHANG ; Aiwei HE ; Pengyue HU ; Ping WANG ; Xiaoyan CHEN
Chinese Journal of Endemiology 2024;43(6):492-497
Objective:To evaluate the application of the standard of "Delimitation and Classification of Keshan Disease Areas" (GB 17020-2010, Standard for short), learn about the applicability of its technical indicators and requirements, and provide a basis for revision of the Standard.Methods:In March 2022, provinces with severe epidemic areas of Keshan disease or new cases reported in recent years were selected, including Gansu Province, Inner Mongolia Autonomous Region, Shandong Province, Shaanxi Province, Yunnan Province, Shanxi Province, Liaoning Province, and Chongqing City. Multi-stage stratified sampling method and questionnaire survey were adopted to collect information on the application of Standard by relevant health institutions.Results:A total of 448 questionnaires were collected, including 445 valid. The survey results showed that 87.64% (390/445) of the respondents were aware of the Standard, and 64.72% (288/445) had received training on the Standard. Eighty-two point two per cent (365/445) of the respondents believed that the Standard was simple and easy to operate, 83.82% (373/445) believed that the determination of the diseased townships was scientific and reasonable, and could be effectively implemented, and 83.60% (372/445) believed that the determination of historical epidemic areas was applicable to the current situation of Keshan disease; 38.88% (173/445), 38.20% (170/445), and 37.98% (169/445) of the respondents believed that the classification indicators for classifying epidemic areas (severe, moderate, and mild epidemic areas) were not applicable to the current situation of the disease in the local or provinces with Keshan disease, respectively. Among the indicators for epidemic areas classification, 30.79% (137/445), 29.21% (130/445), and 28.54% (127/445) of the respondents thought that the annual prevalence, the annual number of new cases and the annual incidence were most suitable for classification of Keshan disease areas.Conclusions:The Standard has been applied well in practice. However, based on the current situation of Keshan disease, it is suggested to redefine the standard for the severity of the disease and the classification of historical epidemic areas.


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