1.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
2.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
3.Anthraquinones of Cassiae Semen alleviate lipid accumulation in obesity by regulating brown adipose tissue and liver function.
Yijie LI ; Ruiyu WU ; Xin LI ; Jianan LI ; Yinhao ZHANG ; Yanbo HUANG ; Guifang FAN ; Xiaojiaoyang LI
Chinese Herbal Medicines 2025;17(3):488-499
OBJECTIVE:
Cassiae Semen (CS, Juemingzi in Chinese) is a widely used traditional Chinese medicine with a variety of pharmacological effects. This study aimed to investigate the potential therapeutic effects and molecular mechanisms of anthraquinones of CS (AQS) for adiposity.
METHODS:
The chemical components of the AQS were determined using high-performance liquid chromatography (HPLC). Network pharmacology analysis was used to predict potential anti-obesity targets of action for AQS. We constructed high fat with high sugar water diet-induced obese mice and observed their body weight and whole-body lipid metabolism to evaluate the efficacy of AQS in promoting lipid metabolism. Subsequently, the epidermal temperature at the brown adipose tissue (BAT) before and after cold stimulation was observed and the expression of lipid metabolism-related genes in the liver and BAT tissues was detected to clarify the mechanism of action of AQS.
RESULTS:
Network pharmacology analysis showed that AQS was involved in the regulation of liver and adipose tissue function under obesity. Pathological and biochemical results showed that AQS reduced lipid accumulation in the liver and adipose tissue induced by an unhealthy diet. With the increase of cold tolerance, the volume and weight of BAT were increased by AQS, suggesting that it regulated the body heat production dominated by BAT. After AQS treatment, the levels of genes related to uncoupling protein1 (UCP1)-mediated adaptive thermogenesis in BAT tissues and lipid metabolism in the liver were also increased, which further proved that AQS activated BAT function to promote lipid metabolism in the whole body.
CONCLUSION
This study revealed the pharmacological effects of AQS, thereby providing a scientific basis for regulating BAT thermogenesis and liver lipid metabolism to alleviate obesity and providing clues for further exploring the application of natural active ingredients in the treatment of metabolism-related diseases.
4.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.
5.Depressive and anxious symptoms and their comorbidity among Chinese residents in 2021 and 2022
Limin RONG ; Yi ZHENG ; Ximing DUAN ; Yanzhi LIU ; Xiaoyan ZHANG ; Ruiyu HU ; Li ZHU ; Yueqin HUANG ; Yibo WU ; Fuqin MU ; Yan LIU
Chinese Mental Health Journal 2023;37(12):1023-1030
Objective:To analyze the detection rate and related factors of depressive and anxious symptoms comorbidity in 2021 and 2022.Methods:Based on the results of the Seventh National Population Census in 2021,the residents of 32 provinces,municipalities,and autonomous regions were sampled by gender and age.The gender and age of the samples were in line with the characteristics of China's population.A face-to-face interview survey was conducted in community residents in each province in 2021(n=11 005)and 2022(n=30 421)with the Gen-eralized Anxiety Questionnaire-7 and Patient Health Questionnaire-9.Results:The detection rates of depressive and anxious symptoms comorbidity were 10.67%in 2021 and 11.72%in 2022.The prevalence of depressive and anxi-ety comorbidity were higher in male,younger(age≤17 years),divorced,lower BMI(BMI<18.5 kg/m2),higher education(graduate),students,and residents with chronic medical history(Ps<0.001).In 2022,32.06%of people with depressive symptoms had anxious symptoms and 47.62%of people with anxious symptoms had depressive symptoms.Conclusion:In 2021 and 2022,the detection rates of depressive and anxious symptoms comorbidity were both about 10%,and half of patients with anxious symptoms were accompanied by depressive symptoms,So atten-tion should be paid to the comorbidity of depression and anxiety symptoms.
6.Analyze 11425 cases of congenital malformations for birth defects intervention
Ziyu WU ; Yanxia LIANG ; Liqun XIE ; Ruiyu HUANG ; Baolin CHEN ; Qiuyuan HE
The Journal of Practical Medicine 2017;33(10):1694-1696
Objective To collect the incidence and distribution of congenital malformations ,find monitor-ing and preventive measures,to provide a basis for reasonable allocation of health resources for clinical depart-ments. Methods According to The International Classification of Diseases(ICD-10)chapter 17 encoding table, we analyze our hospital′s cases from January 2003 to December 2010 in order to understand the incidence of con-genital malformations of every system. Results Top five diseases are:cleft palate deformity ,genital malforma-tion,congenital heart disease and digestive system malformation in equal third place,and facial deformity. The age ranged from 1d to 35 y,men more than women. Conclusions The key to effectively reduce the incidence of con-genital malformation is strictly implement the three-level intervention mechanism. Prevention ,early detection and early treatment will improve the patient′s quality of life.
7.Clinical value of 1.5T MRI DWI and SWI sequence in the diagnosis of cerebral infarction and cerebral hemorrhage
Ruiyu HUANG ; Xia YU ; Baogang XU ; Xingzhi MA ; Jian WANG
Journal of Clinical Medicine in Practice 2017;21(13):92-94
Objective To analyze the clinical value of 1.5T MRI DWI and SWI sequence in the diagnosis of cerebral infarction and cerebral hemorrhage.Methods A total of 28 patients with cerebral infarction were selected as control group,and 28 patients with cerebral hemorrhage were selected as observation group.The PHILPS 1.5T magnetic resonance imaging system was applied.The patients were examined by T1WI and T2WI sequence scanning,and DWI and SWI sequence images were obtained.In the two groups,lesions around the center and different characteristics of SWI and DWI were observed and compared,and the measurement of peripheral lesions were measured for calculating the central area of the PV and ADC.Results There was no significant difference between T1WI and T2WI in patients with cerebral hemorrhage and cerebral infarction,but there was significant difference between DWI sequence and DWI sequence.MRI routine examination showed that there were no significant differences in detection rates of T1WI sequence and T2WI sequence between two groups,but the detection rate of DWI sequence in the observation group was significantly lower than the control group,while detection rate of DWI sequence was significantly higher than the control group(P<0.05).The central area of ADC in the observation group was significantly better than that in the control group,and the PV value of the hematoma area was significantly lower than that of the control group(P<0.05).Conclusion Collections of DWI and SWI images of cerebral infarction and cerebral hemorrhage by magnetic resonance prove that there are significant differences between characteristics of two images.
8.Clinical value of 1.5T MRI DWI and SWI sequence in the diagnosis of cerebral infarction and cerebral hemorrhage
Ruiyu HUANG ; Xia YU ; Baogang XU ; Xingzhi MA ; Jian WANG
Journal of Clinical Medicine in Practice 2017;21(13):92-94
Objective To analyze the clinical value of 1.5T MRI DWI and SWI sequence in the diagnosis of cerebral infarction and cerebral hemorrhage.Methods A total of 28 patients with cerebral infarction were selected as control group,and 28 patients with cerebral hemorrhage were selected as observation group.The PHILPS 1.5T magnetic resonance imaging system was applied.The patients were examined by T1WI and T2WI sequence scanning,and DWI and SWI sequence images were obtained.In the two groups,lesions around the center and different characteristics of SWI and DWI were observed and compared,and the measurement of peripheral lesions were measured for calculating the central area of the PV and ADC.Results There was no significant difference between T1WI and T2WI in patients with cerebral hemorrhage and cerebral infarction,but there was significant difference between DWI sequence and DWI sequence.MRI routine examination showed that there were no significant differences in detection rates of T1WI sequence and T2WI sequence between two groups,but the detection rate of DWI sequence in the observation group was significantly lower than the control group,while detection rate of DWI sequence was significantly higher than the control group(P<0.05).The central area of ADC in the observation group was significantly better than that in the control group,and the PV value of the hematoma area was significantly lower than that of the control group(P<0.05).Conclusion Collections of DWI and SWI images of cerebral infarction and cerebral hemorrhage by magnetic resonance prove that there are significant differences between characteristics of two images.
9.Study on the efficacy and safety of sublingual immunotherapy with standardized dermatophagoides farinae drops for allergic rhinitis.
Shaopeng HUANG ; Xiufang XIE ; Yong CHEN ; Lihua WU ; Ruiyu LI ; Fan SHEN ; Hong CHEN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(7):618-621
OBJECTIVE:
To evaluate the efficacy and safety of the sublingual immunotherapy with dermatophagoides farinae drops on patients with allergic rhinitis.
METHOD:
One hundred and twelve cases were collected from adult patients with dust-mite allergic rhinitis of our hospital who could adhere to treatment and regular follow-up. These patients were randomly allocated to receive either sublingual immunotherapy (SLIT group, n = 56) or medical treatment (Control group, n = 56). To evaluate the clinical efficacy by side effects which were registered, symptom and medication scores which were assessed and rhinoconjunctivitis quality of life questionnaire (RQLQ) which was completed in the baseline and two years after treatment.
RESULT:
Dropouts after the 2 years' treatment were 5 of SLIT group and 4 of Control group respectively. SLIT group induced the significant reductions on both the symptom scores (7.81 ± 3.14 to 3.89 ± 2.01, P < 0.0 1) and the medication scores (2.86 ± 0.75 to 0.44 ± 0.06, P < 0.01). Meanwhile, Control group induced the reductions on both the symptom scores (8.01 ± 3.32 to 5.20 ± 2.43) and the medication scores (2.95 ± 0.80 to 1.75 ± 0.40). There were significant differences (P< 0. 01) in symptom and medication scores between the two groups after 2-year treatment. The patients in SLIT group had fewer symptoms and lower intake of medication. There were statistically significant differences in RQLQ between SLIT group [19 (15,22)] and Control group [36 (26,47)] after two years treatment (Z = -5. 21, P < 0.01). SLIT group also had significant improvement in RQLQ (Z = -6.10, P < 0.01) between before and after the treatment. There were 4 patients who showed adverse reactions in SLIT group (3 occurred in increment period, and 1 occurred in the maintenance period). The incidence of adverse reactions was 7.14%. No severe systemic side effects were registered.
CONCLUSION
SLIT with standardized dermatophagoides farinae drops in China is safe and effective to patients with allergic rhinitis.
Administration, Sublingual
;
Adult
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Animals
;
Antigens, Dermatophagoides
;
immunology
;
China
;
Dermatophagoides farinae
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Humans
;
Quality of Life
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Rhinitis, Allergic
;
drug therapy
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Sublingual Immunotherapy
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Treatment Outcome
10.320 slice CT electrocardiograph-gating double phase cardiac function angiography scan mode:a preliminary investigation of one-stop-shop exam
Xiaoyong HUANG ; Zixu YAN ; Zhaoqi ZHANG ; Xin PU ; Ruiyu DOU ; Hong JIANG ; Miao GUO ; Yi LIU ; Miaomiao JI
Chinese Journal of Radiology 2011;45(1):22-25
Objective To explore the feasibility of evaluating cardiac structure, coronary artery,pulmonary artery and cardiac function in one single scan by 320-row CT ECG-gated double phase cardiac function scan mode. MethodsForty patients underwent the 320-detector row CT double phase cardiovascular angiography. The pulmonary phase and aortic phase were reconstructed in order to evaluate the pulmonary and coronary artery. MPR reconstructions of both pulmonary and aortic phase were used to analyze the function of the two ventricles. And the results of the cardiac function were compared with those of transthoracic echocardiography. Thirty-five cases could be analyzed and diagnosed, while the other 5 cases had to be given up because of the poor imaging quality. The mean heart rate was (71.2 ± 11.2) beat per min (bpm). No arrhythmia case included. Results ( 1 ) Pulmonary embolism were diagnosed in 11 cases,coronary artery disease (CAD) were found in 5 cases, while post-stent implantation were observed in 7 cases. Six cases of congenital heart disease were diagnosed with 3 ASD and 3 primary pulmonary hypertension. Another one was diagnosed with left atrial myxoma, and 5 cases were pulmonary embolism associated with CAD. All of above cases were verified by final clinical diagnosis. (2) The heart function parameters including LVEDd , RVEDd, LVESd, RVESd and LVEF were (36.7 ±3.3), (43.3 ± 3.4) mm,(31.6±5.1), (41.3 ±5.1) mm and (47.1 ±15.1) for CT, while those were (40.3 ±3.1), (47.3 ±4.2) mm,(37.3 ±5.6), (45.3 ±3.3) mm,and (46.0 ± 14.8) for ultrasound, respectively. The CT results were correlated with the ultrasound ( n = 35, r = 0.886-0.988, P < 0.01 ). (3) The average radiation exposure was ( 5.4 ± 0.5 ) mSv. Conclusions 320-row CT ECG-gated double phase cardiac function scan mode is feasible for the "one-stop-shop" examination of the cardiovascular disease. This noninvasive method is recommended for the diagnosis, differential diagnosis, treatment and prognosis of cardiovascular disease.

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