1.Astragali Radix Polysaccharides Promote M2 Polarization of OGD/R-induced BV2 Microglia by Inhibiting TLR4/NF-κB Signaling Pathway
Yanxi LIU ; Lijun ZHANG ; Qiule LI ; Yayu ZENG ; Yanjie HUO ; Xiaodan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):133-143
ObjectiveTo investigate the effects of Astragali Radix polysaccharides (APS) on the polarization of BV2 microglial cells in an oxygen-glucose deprivation/reoxygenation (OGD/R) model through regulation of the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway. MethodsThe OGD/R injury model of BV2 microglia was established and divided into blank group, OGD/R group and APS group (0.4 g·L-1 APS). Neuroinflammatory injury was induced by lipopolysaccharide (LPS) and treated with APS. The cells were divided into blank group, LPS group (1 mg·L-1 LPS) and APS group (0.4 g·L-1 APS+1 mg·L-1 LPS). Cell viability was detected using the cell counting kit-8 (CCK-8) assay. Cell morphology was observed under an inverted microscope. Nitric oxide (NO) content in the cell supernatant was determined by the Griess assay. The secretion levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-10, and IL-4 were measured by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence (IF) was used to detect the double-positive rates of ionized calcium-binding adapter molecule-1/inducible nitric oxide synthase (Iba-1+/iNOS+) and ionized calcium-binding adapter molecule-1/arginase 1 (Iba-1+/Arg1+), as well as the nuclear translocation rate of nuclear factor-κB p65 (NF-κB p65). Protein expression levels of Iba-1, iNOS, Arg1, TLR4, and NF-κB p65 were detected by Western blot. ResultsIn the OGD/R injury model, compared with the blank control group, BV2 microglial cells in the OGD/R group were activated and exhibited amoeboid morphological changes. The secretion levels of NO, TNF-α, and IL-6 were significantly increased (P<0.01). The double-positive expression rate of Iba-1+/iNOS+ and the protein expression of Iba-1 and iNOS were significantly increased (P<0.01). The nuclear translocation rate of NF-κB p65 and the protein expression levels of TLR4 and NF-κB p65 were significantly increased (P<0.01). The levels of IL-10 and IL-4 were significantly decreased (P<0.01), and the double-positive expression rate of Iba-1+/Arg1+ and Arg1 protein expression were significantly decreased (P<0.01). Compared with the OGD/R group, the APS group (0.4 g·L-1) showed reduced cell activation, significantly decreased secretion levels of NO, TNF-α, and IL-6 (P<0.01), significantly decreased double-positive expression rate of Iba-1+/iNOS+ and relative protein expression of Iba-1 and iNOS (P<0.01), significantly decreased nuclear translocation rate of NF-κB p65 and protein expression levels of TLR4 and NF-κB p65 (P<0.01), significantly increased levels of IL-10 and IL-4 (P<0.01), and significantly increased double-positive expression rate of Iba-1+/Arg1+ and Arg1 protein expression (P<0.01). In the LPS-induced neuroinflammation model, compared with the blank control group, the LPS group showed increased cell activation, significantly increased levels of NO, TNF-α, and IL-6, significantly increased Iba-1+/iNOS+ double-positive expression rate, NF-κB p65 nuclear translocation rate, and protein expression levels of Iba-1, iNOS, TLR4, and NF-κB p65 (P<0.01), while IL-10 and IL-4 levels, Iba-1+/Arg1+ double-positive expression rate, and Arg1 protein expression were significantly decreased (P<0.01). Compared with the LPS group, the APS group showed reduced cell activation, significantly decreased levels of NO, TNF-α, and IL-6, Iba-1+/iNOS+ double-positive expression rate, NF-κB p65 nuclear translocation rate, and protein expression levels of Iba-1, iNOS, TLR4, and NF-κB p65 (P<0.01), while IL-10 and IL-4 levels, Iba-1+/Arg1+ double-positive expression rate, and Arg1 protein expression were significantly increased (P<0.01). ConclusionAPS may reduce microglial activation and promote their polarization toward the M2 phenotype by inhibiting activation of the TLR4/NF-κB signaling pathway, thereby alleviating the neuroinflammatory response induced by OGD/R.
2.Academic Characteristics of Contemporary Chinese Medicine Masters in Treating Diabetic Kidney Disease Based on SrTO
Yu SUN ; Xiaodan WANG ; Yingzi CUI ; Tianying CHANG ; Fan LI ; Lisha WANG ; Chenxuan DONG ; Shoulin ZHANG ; Xing LIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):258-269
ObjectiveTo explore the academic characteristics of contemporary renowned Chinese medicine masters in treating diabetic kidney disease (DKD) from the perspectives of principles, methods, formulas, and medications. MethodsIn strict accordance with the Systematic Review of Text and Opinion (SrTO) process developed by the Joanna Briggs Institute (JBI), an Australian evidence-based healthcare center, the databases including China National Knowledge Infrastructure (CNKI), VIP Database, Wanfang Data, and China Biomedical Literature Service System (SinoMed) were searched. Based on predefined inclusion and exclusion criteria, text information extraction, quality evaluation, and text information synthesis were conducted sequentially. The data were analyzed and presented in the form of text and figures. ResultsA total of 215 articles related to 43 contemporary renowned experts in the fields of Chinese medicine nephrology and endocrinology were included. The study found that the academic thoughts of these masters in the treatment of DKD are extensive, involving multiple levels such as disease understanding, therapeutic strategies, formula application, and medication use. In terms of disease understanding, the primary pathogenesis is characterized by deficiency in the root and excess in the manifestation. It is emphasized that internal factors, such as congenital endowment deficiency, interact with external factors such as improper diet, emotional disturbances, invasion of exogenous pathogens, and delayed or inappropriate treatment, to jointly induce the disease. This further gives rise to various pathogenetic theories, including obstruction of renal collaterals by blood stasis, toxin-induced damage to renal collaterals, latent wind disturbing the kidney, and internal heat leading to mass formation. In terms of therapeutic strategies and medication use, the principal treatment method is to replenish Qi and nourish Yin. Stage-based and syndrome-differentiated treatments are advocated. Flexible use of insect-derived drugs and wind-dispelling drugs is emphasized, along with proficiency in applying classical formulas and drug pairs. Integrated internal and external treatments, as well as the combined application of multiple therapeutic approaches, are commonly employed for comprehensive management. Meanwhile, the concept of "preventive treatment of disease" is upheld, and individualized long-term management of patients is advocated. ConclusionThrough the SrTO process, the academic thoughts of contemporary renowned Chinese medicine masters in the treatment of DKD have been systematically and standardly synthesized, providing a scientific and standardized basis for future theoretical exploration.
3.Academic Characteristics of Contemporary Chinese Medicine Masters in Treating Diabetic Kidney Disease Based on SrTO
Yu SUN ; Xiaodan WANG ; Yingzi CUI ; Tianying CHANG ; Fan LI ; Lisha WANG ; Chenxuan DONG ; Shoulin ZHANG ; Xing LIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):258-269
ObjectiveTo explore the academic characteristics of contemporary renowned Chinese medicine masters in treating diabetic kidney disease (DKD) from the perspectives of principles, methods, formulas, and medications. MethodsIn strict accordance with the Systematic Review of Text and Opinion (SrTO) process developed by the Joanna Briggs Institute (JBI), an Australian evidence-based healthcare center, the databases including China National Knowledge Infrastructure (CNKI), VIP Database, Wanfang Data, and China Biomedical Literature Service System (SinoMed) were searched. Based on predefined inclusion and exclusion criteria, text information extraction, quality evaluation, and text information synthesis were conducted sequentially. The data were analyzed and presented in the form of text and figures. ResultsA total of 215 articles related to 43 contemporary renowned experts in the fields of Chinese medicine nephrology and endocrinology were included. The study found that the academic thoughts of these masters in the treatment of DKD are extensive, involving multiple levels such as disease understanding, therapeutic strategies, formula application, and medication use. In terms of disease understanding, the primary pathogenesis is characterized by deficiency in the root and excess in the manifestation. It is emphasized that internal factors, such as congenital endowment deficiency, interact with external factors such as improper diet, emotional disturbances, invasion of exogenous pathogens, and delayed or inappropriate treatment, to jointly induce the disease. This further gives rise to various pathogenetic theories, including obstruction of renal collaterals by blood stasis, toxin-induced damage to renal collaterals, latent wind disturbing the kidney, and internal heat leading to mass formation. In terms of therapeutic strategies and medication use, the principal treatment method is to replenish Qi and nourish Yin. Stage-based and syndrome-differentiated treatments are advocated. Flexible use of insect-derived drugs and wind-dispelling drugs is emphasized, along with proficiency in applying classical formulas and drug pairs. Integrated internal and external treatments, as well as the combined application of multiple therapeutic approaches, are commonly employed for comprehensive management. Meanwhile, the concept of "preventive treatment of disease" is upheld, and individualized long-term management of patients is advocated. ConclusionThrough the SrTO process, the academic thoughts of contemporary renowned Chinese medicine masters in the treatment of DKD have been systematically and standardly synthesized, providing a scientific and standardized basis for future theoretical exploration.
4.Relationship between skin failure and nutritional status in elderly critically ill patients and its predictive efficiency
Bailian LI ; Jinchun GUO ; Xiaodan HAO ; Jiao DU
Journal of Public Health and Preventive Medicine 2026;37(3):172-175
Objective To investigate the relationship of skin failure (SF) with nutritional status in elderly critically ill patients and analyze the predictive efficiency of nutritional status on SF. Methods A total of 340 elderly critically ill patients admitted to the hospital from January 2020 to January 2025 were selected as research subjects. According to whether skin failure occurred, the above patients were classified into skin failure group and non-skin failure group. The nutritional status indicators [serum albumin (ALB), prealbumin (PA), total protein (TP), hemoglobin (Hb), body mass index (BMI), nutritional risk score (NRS 2002)] were compared between both groups. Multivariate analysis was performed on statistically significant indicators, and ROC curve was applied to analyze the predictive value. Results Among the 340 elderly critically ill patients, 142 cases (41.76%) developed skin failure. The ALB, PA, TP, Hb and BMI in the skin failure group were lower than those in the non-skin failure group (P<0.05) while the NRS2002 score was higher (P<0.05). After logistic multivariate analysis, ALB, PA and Hb were independent influencing factors of skin failure (P<0.05). ROC curve analysis revealed that the predictive value of ALB (AUC=0.850, 95%CI: 0.808-0.888, Z=-3.707, P<0.001) was better than that of PA (AUC=0.770, 95%CI: 0.717-0.816, Z=-3.100, P=0.002) or Hb (AUC=0.773, 95%CI: 0.722-0.819, Z=-2.556, P=0.011). Conclusion The occurrence of skin failure in elderly critically ill patients is closely related to nutritional status. ALB, PA and Hb are independent risk factors of SF, and ALB has the best predictive efficiency on SF.
5.Effect evaluation of short video based health education on e-cigarette awareness improvement among junior high school students
LI Da,WANG Xiaodan,LI Lei,LIU Na,ZHAO Lei
Chinese Journal of School Health 2026;47(6):799-803
Objective:
To evaluate the efficacy of a short video based health education model in enhancing e-cigarette awareness among junior high school students, so as to provide empirical evidence for optimizing school based health education in the digital era.
Methods:
In December 2024, a cluster sampling method was applied to recruit 465 eighth grade students from a middle school in Kunming for a survey. The intervention was implemented through playing short videos produced by the research team in the classroom, and a self designed adolescent e-cigarette questionnaire was used to conduct assessments before and after intervention, along with qualitative interviews with 60 teachers and students by purposive sampling. Pre and post intervention outcomes were compared by using paired t-test and Chi-square test. Qualitative data underwent a three stage coding process of "open, axial, and selective coding" to construct an explanatory framework.
Results:
Quantitative analysis showed significant post intervention increases in both baseline awareness scores(92.1±6.6)and deep awareness scores (70.0±17.6) compared with pre intervention( 78.2± 14.0,46.1±20.5)( t =23.54,26.72,both P <0.05), both meeting the criteria for large effect sizes (Cohen s d =1.09, 1.24). The improvement in deep awareness was particularly pronounced, specifically regarding addictive ingredients, illicit additives(tiletamine,metomidate,etomidate), and misconceptions about smoking cessation efficacy ( χ 2=150.54,113.27,117.40,112.92, 69.93 , all P <0.01). Qualitative insights revealed that narrative transportation and visual enhancement were the core mechanisms of efficacy: 87.3% of students preferred contextualized narratives, 85.5% of students decoded cognitive bottlenecks more effectively because of visual enhancement, and 72.7% relied on short videos for micro learning.
Conclusion
Short video based health education can effectively enhance e-cigarette awareness among junior high school students, offering a pragmatically viable digital pathway for school based health education.
6.Research progress of perfluorohexyloctane eyedrop in meibomian gland dysfunction-associated dry eye
Hongyue YU ; Bin SUN ; Yan LI ; Xiaodan LI ; Meng XIN
International Eye Science 2026;26(8):1354-1358
Dry eye disease(DED)is a multifactorial ocular surface disorder characterized by the loss of tear film homeostasis among which evaporative DED associated with meibomian gland dysfunction(MGD)accounts for up to 86%, and traditional drugs fail to effectively target the core pathological mechanism. As a new type of anhydrous and preservative-free eye drops approved by the FDA, perfluorohexyloctane has a unique amphiphilic structure, which can form an artificial lipid monolayer at the air-liquid interface of the tear film. It exerts its therapeutic effects by inhibiting tear evaporation, stabilizing the tear film, unclogging meibomian gland orifices, and promoting corneal oxygen supply. This drug can not only serve as a first-line treatment option for evaporative DED but also be combined with other drugs for personalized treatment. This review summarizes the mechanism of action, pharmacological properties, clinical efficacy, and safety of perfluorohexyloctane eye drops, and further elaborate on its application value in complex types of DED and patients with other concurrent ocular surface diseases, aiming to provide more evidence support for the precise treatment of DED.
7.Research progress of degenerative retinoschisis
Yaqi BAI ; Jing LI ; Xiaodan ZHANG ; Wenjuan WANG ; Yongrui WANG ; Yanan QIN ; Jiajia YANG ; Guohong ZHOU
International Eye Science 2026;26(9):1566-1571
Degenerative retinoschisis(RS)is a common peripheral retinal degeneration that typically remains asymptomatic and follows a benign clinical course. However, its morphological resemblance to rhegmatogenous retinal detachment(RRD)presents a diagnostic challenge, as the two conditions demand fundamentally different management and carry distinct prognoses. Precise differentiation is therefore clinically imperative. Recent advances in multimodal imaging have substantially refined the diagnostic accuracy and differential assessment of degenerative RS, establishing spectral-domain optical coherence tomography(SD-OCT)as the definitive reference standard. When the disease progresses to progressive symptomatic schisis-related retinal detachment(PSSRD), prompt surgical intervention, including scleral buckling, pars plana vitrectomy, or a combined approach, is mandated to preserve visual function. Clinical decision-making must be individualized, with the surgical strategy carefully tailored to the specific characteristics of eachcase. This review provides a systematic synthesis of current evidence on the natural history, imaging hallmarks, and surgical management of degenerative RS and its complications, aiming to inform and optimize clinical practice.
8.Randomized Controlled Study on Wenshen Yangxue Decoction Plus Endometrial Microstimulation for Polycystic Ovary Syndrome (PCOS) Infertility with Syndrome of Kidney-Yang Deficiency and Blood Stasis
Qianqian HUANG ; Qian HAN ; Mingwei XIN ; Junqin HE ; Jingshang WANG ; Xiaodan YIN ; Mengyuan LI ; Yanxiao YI ; Yanli TANG ; Yiting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):181-190
ObjectiveTo evaluate the clinical efficacy and safety of Wenshen Yangxue decoction combined with endometrial microstimulation in the treatment of infertile patients with polycystic ovary syndrome (PCOS, syndrome of kidney-Yang deficiency and blood stasis). MethodsA randomized, positive-drug parallel-controlled, open-label clinical study was conducted. A total of 240 infertile PCOS patients with the syndrome of kidney-Yang deficiency and blood stasis who met the inclusion and exclusion criteria were randomly assigned into 4 groups, with 60 patients in each group. The control group was treated with clomiphene citrate (50-100 mg·d-¹, orally for 5-10 consecutive days) for ovulation induction. On the basis of the therapy in the control group, the Wenshen group was additionally treated with Wenshen Yangxue decoction (250 mL, warm oral administration, twice daily, in the morning and evening), and the stimulation group received endometrial microstimulation during the early follicular phase. The combined group was treated with clomiphene citrate plus Wenshen Yangxue decoction (at the same dosages as aforementioned) and endometrial microstimulation. The course of treatment for all the groups was 3 menstrual cycles. The pregnancy rate, ovulation status, sex hormone levels, glucose metabolism indicators, coagulation function, endometrial thickness, traditional Chinese medicine (TCM) symptom scores, and adverse reactions were observed in each group. ResultsA total of 239 patients were included in the final analysis, with 59 patients in the Wenshen group and 60 patients in each of the other three groups. The pregnancy rates were 43.3%(26/60) in the combined group and 32.2%(19/60) in the Wenshen group, both significantly higher than that (10%) in the control group (χ2=17.56,P<0.01). Compared with the control group, the combined and Wenshen groups had increased mature follicle rates (P<0.01), while the Wenshen group showed a decreased luteinization rate (P<0.05). All the groups exhibited significant reductions in TCM symptom scores after treatment, and the Wenshen and combined groups had lower scores than the control group (P<0.01). No statistically significant difference was found in the incidence of adverse events among the four groups, and all the adverse events were mild. ConclusionWenshen Yangxue decoction combined with endometrial microstimulation can alleviate the TCM symptoms of infertile PCOS patients with the syndrome of kidney-Yang deficiency and blood stasis, promote follicular maturation and ovulation, increase the cumulative pregnancy rate, and has good safety. It can provide new clinical evidence for the integrated traditional Chinese and Western medicine treatment of PCOS infertility.
9.Randomized Controlled Study on Wenshen Yangxue Decoction Plus Endometrial Microstimulation for Polycystic Ovary Syndrome (PCOS) Infertility with Syndrome of Kidney-Yang Deficiency and Blood Stasis
Qianqian HUANG ; Qian HAN ; Mingwei XIN ; Junqin HE ; Jingshang WANG ; Xiaodan YIN ; Mengyuan LI ; Yanxiao YI ; Yanli TANG ; Yiting WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):181-190
ObjectiveTo evaluate the clinical efficacy and safety of Wenshen Yangxue decoction combined with endometrial microstimulation in the treatment of infertile patients with polycystic ovary syndrome (PCOS, syndrome of kidney-Yang deficiency and blood stasis). MethodsA randomized, positive-drug parallel-controlled, open-label clinical study was conducted. A total of 240 infertile PCOS patients with the syndrome of kidney-Yang deficiency and blood stasis who met the inclusion and exclusion criteria were randomly assigned into 4 groups, with 60 patients in each group. The control group was treated with clomiphene citrate (50-100 mg·d-¹, orally for 5-10 consecutive days) for ovulation induction. On the basis of the therapy in the control group, the Wenshen group was additionally treated with Wenshen Yangxue decoction (250 mL, warm oral administration, twice daily, in the morning and evening), and the stimulation group received endometrial microstimulation during the early follicular phase. The combined group was treated with clomiphene citrate plus Wenshen Yangxue decoction (at the same dosages as aforementioned) and endometrial microstimulation. The course of treatment for all the groups was 3 menstrual cycles. The pregnancy rate, ovulation status, sex hormone levels, glucose metabolism indicators, coagulation function, endometrial thickness, traditional Chinese medicine (TCM) symptom scores, and adverse reactions were observed in each group. ResultsA total of 239 patients were included in the final analysis, with 59 patients in the Wenshen group and 60 patients in each of the other three groups. The pregnancy rates were 43.3%(26/60) in the combined group and 32.2%(19/60) in the Wenshen group, both significantly higher than that (10%) in the control group (χ2=17.56,P<0.01). Compared with the control group, the combined and Wenshen groups had increased mature follicle rates (P<0.01), while the Wenshen group showed a decreased luteinization rate (P<0.05). All the groups exhibited significant reductions in TCM symptom scores after treatment, and the Wenshen and combined groups had lower scores than the control group (P<0.01). No statistically significant difference was found in the incidence of adverse events among the four groups, and all the adverse events were mild. ConclusionWenshen Yangxue decoction combined with endometrial microstimulation can alleviate the TCM symptoms of infertile PCOS patients with the syndrome of kidney-Yang deficiency and blood stasis, promote follicular maturation and ovulation, increase the cumulative pregnancy rate, and has good safety. It can provide new clinical evidence for the integrated traditional Chinese and Western medicine treatment of PCOS infertility.
10.Analysis of clinical characteristics and diagnostic prediction of Qi deficiency and blood stasis syndrome in acute ischemic stroke
Hao XU ; Xu ZHU ; Bo LI ; Xiaodan LIU ; Xihui PAN ; Changqing DENG
Digital Chinese Medicine 2025;8(1):111-122
[Objective] :
To explore the clinical characteristics and methods for syndrome differentiation prediction, as well as to construct a predictive model for Qi deficiency and blood stasis syndrome in patients with acute ischemic stroke (AIS).
[Methods] :
This study employed a retrospective case-control design to analyze patients with AIS who received inpatient treatment at the Neurology Department of The First Hospital of Hunan University of Chinese Medicine from January 1, 2013 to December 31, 2022. AIS patients meeting the diagnostic criteria for Qi deficiency and blood stasis syndrome were stratified into case group, while those without Qi deficiency and blood stasis syndrome were stratified into control group. The demographic characteristics (age and gender), clinical parameters [time from onset to admission, National Institutes of Health Stroke Scale (NIHSS) score, and blood pressure], past medical history, traditional Chinese medicine (TCM) diagnostic characteristics (tongue and pulse), neurological symptoms and signs, imaging findings [magnetic resonance imaging-diffusion weighted imaging (MRI-DWI)], and biochemical indicators of the two groups were collected and compared. The indicators with statistical difference (P < 0.05) in univariate analysis were included in multivariate logistic regression analysis to evaluate their predictive value for the diagnosis of Qi deficiency and blood stasis syndrome, and the predictive model was constructed by receiver operating characteristic (ROC) curve analysis.
[Results] :
The study included 1 035 AIS patients, with 404 cases in case group and 631 cases in control group. Compared with control group, patients in case group were significantly older, had extended onset-to-admission time, lower diastolic blood pressure, and lower NIHSS scores (P < 0.05). Case group showed lower incidence of hypertension history (P < 0.05). Regarding tongue and pulse characteristics, pale and dark tongue colors, white tongue coating, fine pulse, astringent pulse, and sinking pulse were more common in case group. Imaging examinations demonstrated higher proportions of centrum semiovale infarction, cerebral atrophy, and vertebral artery stenosis in case group (P < 0.05). Among biochemical indicators, case group showed higher proportions of elevated fasting blood glucose and glycated hemoglobin (HbA1c), while lower proportions of elevated white blood cell count, reduced hemoglobin, and reduced high-density lipoprotein cholesterol (HDL-C) (P < 0.05). Multivariate logistic regression analysis identified significant predictors for Qi deficiency and blood stasis syndrome including: fine pulse [odds ratio (OR) = 4.38], astringent pulse (OR = 3.67), superficial sensory abnormalities (OR = 1.86), centrum semiovale infarction (OR = 1.57), cerebral atrophy (OR = 1.55), vertebral artery stenosis (OR = 1.62), and elevated HbA1c (OR = 3.52). The ROC curve analysis of the comprehensive prediction model yielded an area under the curve (AUC) of 0.878 [95% confidence interval (CI) = 0.855 – 0.900].
[Conclusion]
This study finds out that Qi deficiency and blood stasis syndrome represents one of the primary types of AIS. Fine pulse, astringent pulse, superficial sensory abnormalities, centrum semiovale infarction, cerebral atrophy, vertebral artery stenosis, elevated blood glucose, elevated HbA1c, pale and dark tongue colors, and white tongue coating are key objective diagnostic indicators for the syndrome differentiation of AIS with Qi deficiency and blood stasis syndrome. Based on these indicators, a syndrome differentiation prediction model has been developed, offering a more objective basis for clinical diagnosis, and help to rapidly identify this syndrome in clinical practice and reduce misdiagnosis and missed diagnosis.


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