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【Objective】 To construct the secretory expression system of insect cells to express the secretory TSHR A subunit protein in the ovarian cells of Spotoma oryzae (sf9). 【Methods】 A recombinant plasmid containing the target protein was constructed, and then the positive bacmid was screened out by the blue and white spots experiment. The verified bacmid was transfected into SF9 insect cells to obtain recombinant baculovirus. The virus was amplified, and the titer level was detected by virus plaque assay. Finally, Western blotting was used to identify the expression of the recombinant protein and optimize the expression conditions. 【Results】 During the construction of the protein expression system, PCR identification and sequencing results confirmed the correctness of the sequences of the recombinant plasmid and the recombinant bacmid. After the transfection of the bacmid, the signs of virus budding were observed in sf9 cells. The virus was collected and amplified. The titer of P1 generation virus was 2×107 pfu/m according to the plaque assay. The recombinant protein was identified by Western blotting and confirmed to be exogenous into the culture medium. The optimal condition for virus infection and protein expression was 72 h after the infection when the multiplicity of infection (MOI) was 1. 【Conclusion】 We constructed an insect cell expression system secreting TSHR 22-289 (55 ku), and the protein could be successfully glycolyzed. This system provides a preliminary basis for the construction and production of its industrial platform and also provides a useful tool for studies on TSHR protein and prevention of GO in the future.
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Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide nowadays. Given the very complex pathogenesis, there are yet no effective pharmacological agents for the treatment of NAFLD. Accumulating evidence has shown that cholesterol metabolism dysregulation, resulting in excess cholesterol accumulation in the liver and subsequent hepatic inflammation and injury, is one of the most important causes for the occurrence and development of NAFLD. This suggests the application of cholesterol metabolism- targeting therapeutic strategy in NAFLD. This review firstly summarized the pathogenesis of NAFLD, then focused on the regulation of cholesterol metabolism and underlying mechanisms through which excess hepatic cholesterol contributed to NAFLD, and reviewed the research progress on cholesterol-lowering agents aimed at maintaining the cholesterol homeostasis to treat NAFLD, which would provide insights for novel strategies to treat NAFLD in both preclinical studies and clinical settings.
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Objective:To compare the efficacy and safety of Tonghua Dongbao′s insulin aspart injection (Rishulin) and NovoRapid (Novo Nordisk) in the treatment of diabetes.Methods:A 26-week, randomized, open-label, parallel-group, positive control drug and non-inferiority trial was conducted in 23 centers in China. A total of 563 diabetes with poor blood glucose control treated with insulin for at least 3 months before were included. The subjects were randomized(stratified block random method) into those receiving Rishulin or NovoRapid at a ratio of 3∶1. Both groups were combined with basal insulin (Lantus). The primary endpoint was the change in glycosylated hemoglobin (HbA1c) from baseline to the end of 24 weeks of treatment.Results:For full analysis set, after 24 weeks of treatment, HbA1c level of Ruishulin group decreased from (8.66±1.28)% to (7.77±1.09)% ( P<0.001), and that of NovoRapid group decreased from (8.47±1.28) % to (7.65±0.97) % ( P<0.001). Treatment difference in HbA1c (NovoRapid group-Ruishulin group) was -0.061% (95% CI -0.320-0.199). HbA1c<7.0% target reacing rates were 24.26% and 21.21% ( P=0.456), and HbA1c<6.5% target reacing rates were 9.65% and 6.82% ( P=0.310) in Ruishulin group and NovoRapid group, repectively. The standard 2 hours postprandial blood glucose (2hPG) in Ruishulin group decreased from (16.23±5.22) mmol/L to (12.65±4.57) mmol/L ( P<0.001), and 2hPG in NovoRapid group decreased from (16.13±5.37) mmol/L to (11.91)±4.21) mmol/L ( P<0.001). The fingertips blood glucose at 7-point of both groups exhibited varying degrees of reduction compared with those at baseline, repectively. Positive ratios of specific antibodies were 31.68% in Ruishulin group and 36.36% in NovoRapid group ( P=0.320). Ratios of negative to positive were 7.43% and 10.61% ( P=0.360), and ratios of positive to negative were 10.40% and 7.58% ( P=0.360) in Ruishulin group and NovoRapid group, respectively. The incidence of hypoglycemia was 60.05% and 55.40% ( P=0.371), and the incidence of adverse events was 76.60% and 77.70% ( P=0.818) in Ruishulin group and NovoRapid group, respectively. Conclusions:Rishulin is not inferior to NovoRapid, and has shown good efficacy and safety. It can be an ideal choice for clinicians in patients with poor blood glucose control with insulin.
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Background@#Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. @*Methods@#Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. @*Results@#The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. @*Conclusion@#The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.
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Background@#Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. @*Methods@#Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. @*Results@#The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. @*Conclusion@#The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.
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Objective@#To study the influences of dihydrotestosterone (DHT) on the development of experimental autoimmune Graves disease (EAGD), and to observe the effect of DHT on cytokines in male BALB/c mice model.@*Methods@#Male BALB/c mice aged 6-8 weeks were divided into 4 groups using random number table: (1) control group; (2) EAGD group; (3) placebo group; (4) DHT group. EAGD mice were induced with an adenovirus expressing the human thyroid stimulating hormone receptor antibody A-subunit (Ad-TSHR289). DHT (5mg) or a matching placebo were implanted one week before the first immunization. Thyroid hormones were detected with radioimmunoassay kit.. Cytokines [such as interferonγ (IFNγ), interleukin (IL)-4, IL-10, IL-9, and IL-17] producing cells from the spleen were detected using flow cytometry.@*Results@#As expected Ad-TSHR289 treatment increased total thyroxine [EAGD group vs. control group: (117.76±32.69) nmol/L vs. (33.08±12.61) nmol/L, P<0.0001] and free thyroxine [EAGD group vs. control group: (15.01±11.55) pmol/L vs. (3.55±1.88) pmol/L, P<0.0001]. Treatment of DHT slightly lowered thyroid hormones [DHT group vs. placebo group: total thyroxine (114.80±44.27) nmol/L vs. (123.17±77.73) nmol/L; free thyroxine (13.48±6.01) pmol/L vs. (14.19±12.65) pmol/L], without significant difference (all P>0.05)]. However, the percentage of IL-10, but not IFN γ, IL-4, IL-9 and IL-17, secreted spleen cells increased in DHT group than in the placebo group [(7.11±3.29)% vs. (3.51±1.36)%, P<0.05].@*Conclusion@#The effects of DHT on thyroid hormone are mild. It might play an immunomodulatory role in the male mouse Graves disease model by up-regulating the cytokine IL-10.
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Objective:To compare the efficacy and safety of Changsulin ? with Lantus ? in treating patients with type 2 diabetes mellitus (T2DM). Methods:This was a phase Ⅲ, multicenter, randomized, open-label, parallel-group, active-controlled clinical trial. A total of 578 participants with T2DM inadequately controlled on oral hypoglycemic agents were randomized 3∶1 to Changsulin ? or Lantus ? treatment for 24 weeks. The efficacy measures included changes in glycosylated hemoglobin (HbA1c), fasting plasma glucose (FPG), 2h postprandial plasma glucose (2hPG), 8-point self-monitoring of blood glucose (SMBG) profiles from baseline, and proportions of subjects achieving targets of HbA1c and FPG. The safety outcomes included rates of hypoglycemia, adverse events (AEs) and anti-insulin glargine antibody. Results:After 24 weeks of treatment, mean HbAlc decreased 1.16% and 1.25%, FPG decreased 3.05 mmol/L and 2.90 mmol/L, 2hPG decreased 2.49 mmol/L and 2.38 mmol/L in Changsulin ? and in Lantus ?, respectively. No significant differences could be viewed in above parameters between the two groups (all P>0.05). There were also no significant differences between Changsulin ? and Lantus ? in 8-point SMBG profiles from baseline and proportions of subjects achieving the targets of HbA1c and FPG (all P>0.05). The rates of total hypoglycemia (38.00% and 39.01% for Changsulin ? and Lantus ?, respectively) and nocturnal hypoglycemia (17.25% and 16.31% for Changsulin ? and Lantus ?, respectively) were similar between the two groups (all P>0.05). Most of the hypoglycemia events were asymptomatic, and no severe hypoglycemia were found in both groups. No differences were observed in rates of AEs (61.77% vs.52.48%) and anti-insulin glargine antibody (after 24 weeks of treatment, 6.91% vs.3.65%) between the two groups (all P>0.05). Conclusions:Changsulin ? shows similar efficacy and safety profiles compared with Lantus ? and Changsulin ? treatment was well tolerated in patients with T2DM.
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Objective:To study the influences of dihydrotestosterone (DHT) on the development of experimental autoimmune Graves disease (EAGD), and to observe the effect of DHT on cytokines in male BALB/c mice model.Methods:Male BALB/c mice aged 6-8 weeks were divided into 4 groups using random number table: (1) control group; (2) EAGD group; (3) placebo group; (4) DHT group. EAGD mice were induced with an adenovirus expressing the human thyroid stimulating hormone receptor antibody A-subunit (Ad-TSHR289). DHT (5mg) or a matching placebo were implanted one week before the first immunization. Thyroid hormones were detected with radioimmunoassay kit.. Cytokines [such as interferonγ (IFNγ), interleukin (IL)-4, IL-10, IL-9, and IL-17] producing cells from the spleen were detected using flow cytometry.Results:As expected Ad-TSHR289 treatment increased total thyroxine [EAGD group vs. control group: (117.76±32.69) nmol/L vs. (33.08±12.61) nmol/L, P<0.0001] and free thyroxine [EAGD group vs. control group: (15.01±11.55) pmol/L vs. (3.55±1.88) pmol/L, P<0.0001]. Treatment of DHT slightly lowered thyroid hormones [DHT group vs. placebo group: total thyroxine (114.80±44.27) nmol/L vs. (123.17±77.73) nmol/L; free thyroxine (13.48±6.01) pmol/L vs. (14.19±12.65) pmol/L], without significant difference (all P>0.05)]. However, the percentage of IL-10, but not IFN γ, IL-4, IL-9 and IL-17, secreted spleen cells increased in DHT group than in the placebo group [(7.11±3.29)% vs. (3.51±1.36)%, P<0.05]. Conclusion:The effects of DHT on thyroid hormone are mild. It might play an immunomodulatory role in the male mouse Graves disease model by up-regulating the cytokine IL-10.
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Objective:To establish the characteristic T cell receptor(TCR) repertoires which could distinguish the Graves′ disease(GD) and thyroid associated ophthalmopathy(TAO) immune repertoire for precision diagnosis.Methods:Fifteen Graves′ disease patients(nine patients without Graves′ ophthalmopathy and six with Graves′ ophthalmopathy)were enrolled in this study, and they were diagnosed less than 2 months and were naive to treatment. Twenty healthy volunteers who were age and sex matched served as controls. The blood samples were collected at diagnosed. RNA was isolated for TCR repertoire sequencing.Results:TCR diversity in GD and TAO patients was significantly lower than that of healthy controls. With TCR high-throughput sequencing data, characteristic TCR databases of GD and TAO, containing 157 and 116 VJ families, was constructed. In these VJ families, both common and unique families of the two diseases existed.Conclusion:GD and TAO have common pathogenesis, but have different dominant T cell clones, which mediate different immune responses. Therefore, the characteristic TCR of GD and TAO patients may be used as a potential molecular marker.
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This is a follow-up study in two patients who were diagnosed with the occult ectopic adrenocorticotropin syndrome ( EAS) and treated with mifepristone in our hospital. The efficacy and adverse reactions of mifepristone in these two patients with EAS were investigated. The patients had significant improvement in clinical manifestations, the plasma adrenocorticotropic hormone ( ACTH) and cortisol levels decreased in patient 1, while patient 2 remained stable during the treatment. Patient 1 with diabetes mellitus stopped hypoglycemic agents, while the dose of antihypertensive drugs decreased in patient 2 with hypertension, and she was treated with surgery and suffered from transient adrenocortical insufficiency after surgical excision of the lesion. Mifepristone is effective and safe in treating EAS. Surgical treatment should be performed once the lesion is found during the follow-up.
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Objective To explore the relationship between different smoking status and hypothyroidism in six iodine-suitable areas of China. Methods A total of 8187 residents were investigated by cluster sampling in six cities, and 7448 residents were included in the survey. The height, weight, waist circumference, and blood pressure were measured by filling out epidemiological questionnaire. Blood samples were collected to detect thyroid stimulating hormone ( TSH) , thyroid peroxidase antibody ( TPOAb) , and thyroglobulin antibody ( TgAb) . Results The mean TSH, TPOAb, and TgAb positive rates in passive smoking and active smoking groups were all lower than those in non-smoking group ( all P<0.01) . In the active smoking group, the TSH value decreased by 0.023 units for every unit increase in smoking index. The positive rates of TgAb and TPOAb in both passive smoking and active smoking groups were lower than those in non-smoking group (all P<0.01). Active and passive smoking reduced the prevalence of hypothyroidism (both P<0.01). Among women, the risks of clinical hypothyroidism and subclinical hypothyroidism were reduced in both active and passive smoking groups. Besides, the risk of subclinical hypothyroidism decreased significantly when the smoking index was more than 70. In male population, the risk of subclinical hypothyroidism in active and passive smoking group decreased. Besides, the risk of clinical hypothyroidism and subclinical hypothyroidism decreased significantly when the smoking index was more than 70. Conclusion Smoking in iodine-suitable areas may reduce TSH level and the positive rates of TPOAb and TgAb.
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Craniopharyngiomas develop from remnant epithelial cells of Rathke's pouch in the sella/parasella region with low histological grade. Clinical manifestations are related to visual impairment, increased intracranial pressure, and hypothalamic/pituitary deficiencies, including vision loss, headache, nausea, growth retardation, and hypogonadism. Despite benign epithelial tumors, craniopharyngiomas are adjacent to critical brain structures including hypothalamus, pituitary stalk, chiasma opticum and have recurrence tendency. Treatment-related complications including hypothalamic/pituitary deficiencies, diabetes insipidus, and hypothalamic obesity are major risk factors for reduced quality of survival and throw out a great challenge to the surgical treatment and the postoperative management. Clinical features of one case craniopharyngiomas with manifestation of severe hypopituitarism are described with the aim of exploring effective therapies.
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According to the training requirement of the biological-psycho-social medical model to the clinical medical students,in order to promote the comprehensive improvement of medical students' professional knowledge,hands-on ability and human qualities,we design the course of introduction to clinical medicine.Through six major functional modules such as basic professional quality,clinical diagnosis basic technology,basic skills related to the operation,the new progress in clinical medicine and technology,clinical nursing and medical relationship,and medical information management,we build up the core content of integrated course of introduction to clinical medical.In this way,the clinical and related basic knowledge and skills are integrated,the clinical course is closely connected with the basic curriculum,the medical and the humanities exchanges.Through this design,the foundation is laid out for the collaborative efforts of the organ-system of integrated curriculum reform.
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Objective@#To investigate the impact factors for central neck lymph node metastases(CLNM) of papillary thyroid carcinoma(PTC).@*Methods@#A total of 498 patients with PTC who underwent total or hemi-thyroidectomy plus central neck lymph node dissection between January 2014 and July 2016 were included. Univariate and multivariate analyses were performed to identify clinicopathological characteristics, thyroid function parameters and US findings that associated with CLNM of PTC. A nomogram was developed to predict the probability of CLNM. The receiver operating characteristic curve(ROC) was used to estimate the efficiency of the nomogram.@*Results@#Among 498 patients, 284 patients were affected by CNLM. The sensitivity and specificity of US in predicting PTC metastasis in the central neck were 31.3% and 88.3%, respectively. Univariate and multivariate analyses showed that gender, age, number and size of suspicious malignant nodules in thyroid, and suspicious lymph node metastasis detected by ultrasonography were independently correlated with CLNM. The ROC showed that the AUC was 0.748, with sensitivity of 80.8%, and specificity of 59.8%.@*Conclusions@#Gender, age, number and size of suspicious malignant nodules in thyroid, suspicious lymph node metastasis were predictive factors for CLNM in patients with PTC. The nomogram developed based on related factors with CLNM is more sensitive than sonographic central neck lymph node features in predicting the probability of CLNM.
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ABSTRACT:Objective To study the effects of weight and anteroposterior diameter-to-transverse diameter ratio on establishing a model of acute myocardial infarction (AMI)in rats without artificial ventilation and changes in left ventricular function after infarction.Methods Healthy SD rats were randomly divided into group A (200-250 g),group B (250 - 300 g),group C (> 300 g),and group D (control group).The left anterior descending (LAD)coronary artery was ligated to establish a model of myocardial infarction under spontaneous breathing condition immediately after thoracic lines were measured.And changes of electrocardiography were recorded after model establishment.At 2 and 4 weeks after AMI,we observed ventricular wall thickness and ventricular wall motion and measured the changes of cardiac function.Histomorphological changes and myocardial ultrastructure of the heart were observed under thoracotomy 2 weeks after operation.The above data were analyzed by SPSS13.0 statistics software.Results ① The first AMI rat model was established successfully after 30 times of experiments, and after 100 times the model’s success rate gradually stabilized at about 83%.② Group B and group C had a higher model success rate than group A (P 0.05).③ There was no association between the rate of rat thoracic line and modeling success rate (P >0.05). ④ Two weeks after thoracotomy,ischemic myocardial color was white,and ventricular wall motion decreased.HE staining revealed that cardiomyocytes disappeared and were replaced by fibrous tissues and collagen.Remnant cardiomyocytes were arranged disorderly and myofibers were fractured,with interstitial damage and hyperplasia of fibrous tissue.Visible muscle cells were sparse and dissolved,the mitochondria had darker staining,blurred cristae, and edema under electron microscopy. ⑤ Compared with group D, 2 weeks and 4 weeks after myocardial infarction,left ventricular end-diastolic diameter (LVEDD)and left ventricular end systolic diameter (LVEDs) increased (P <0.05),but EF values and heart rate dropped (P <0.05).Conclusion By this method,a model of AMI in rats can be established successfully and the heart function is changed.Under the condition of non-artificial ventilation,the weight of rats is an important factor for establishing AMI model.However,we have not confirmed the effect of thoracic lines on establishing AMI model yet.
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[Summary] The effects of selenium on human health and the related researchs of the impacts of selenium on thyroid were introduced at home and abroad in recent years. The selenium deficiency was correlated with incidence of subclinical and clinical hypothyroidism and autoimmune thyroiditis. Supplement of selenium can improve immune status and thyroid function in patients with autoimmune thyroiditis. There is no definitive evidence for the influence of selenium on Graves disease.
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Objective BRAFV600E mutation, RET/PTC rearrangement, and the concomitant of Hashimoto's thyroiditis(HT) could influence clinicopathological features of papillary thyroid carcinoma (PTC).This study is to investigate the distribution of three factors in PTC and to analyze their associations with clinicopathological characteristics.Methods Fine-needle aspiration samples were collected in a total of 122 conventional PTC patients, who were confirmed by surgery.The clinicopathological features were collected to analyze its association with different factors.BRAFV600E mutation and RET/PTC rearrangement were detected by pyrosequencing and Taqman-qPCR, respectively.Results BRAFV600E mutation was significantly correlated with an older age and a less coexistence with HT(P＜0.05).In contrast, RET/PTC rearrangement was more prevalent in young patients and was associated with the concomitant of HT(P＜0.05).In the age of ≥20 year and＜45 year groups, BRAFV600E mutation was significantly associated with extrathyroidal invasiveness.RET/PTC rearrangement was significantly associated with bilateral lymph node metastasis and the number of metastatic lymph node.Conclusions The distribution of three factors were different in PTC patients.In addition to the age at diagnosis, all of three factors should also be considered together to analyze the association of clinicopathological features of PTC.
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Thyroid hormone insensitivity syndrome is an inherited syndrome characterized by high serum concentrations of thyroid hormone,accompanied by normal or slightly high serum TSH concentration due to the reduced responsiveness of target tissues to thyroid hormone.Thyroid hormone insensitivity syndromes are related to gene mutations of thyroid hormone receptors,thyroid hormone cell transport defect,and thyroid hormone metabolism defect.This article reviews the progress of pathogenesis,diagnosis,and therapeutics of thyroid hormone insensitivity syndrome.
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Private cloud is an internal cloud featuring multi-tenant,dynamic configuration and optimization infrastructure,which enables developers to achieve service self-deployment and self-hosting within security coverage of the enterprise data center.This paper introduced the concept of cloud computing.Then it went on to present the private cloud architecture of the hospital by analysis of problems in the hospital including information construction costs,management and maintenance,and information expansion.In the end,the authors analyzed the cloud computing service model,hospital private cloud architecture,along with outcome analysis for hospital private cloud implementations.