This study was aimed to explore the method to improve the purity of primary myocardial cells from neonatal rats. The myocardial tissues of rats which were born 1-3 days were repeated digested by 0.08% myocardial cells digestive juices. And then, cells were neutralized with DMEM medium which containing 10% fetal bovine serum. The cells were separated with differential sidewall ion. The red blood cell cracking liquid was added to remove the red blood cells. Bromine deoxidization uracil (Brdu) was added to purify the myocardial cells. Then, cells were incubated in the CO2 incubator for two days. The serum-free synchronization was performed for one day. The results showed that the output of myocardial cells from one rat was about 1.2 × 106. The dynamic myocardial cells occupied more than 90%. The purity of myocardial cells was more than 90%. After 3 to 4 days, the cell fusion of myocardial cells was formed with spontaneous rhythm beats. It was concluded that the method can ensure the yield and the activity of the myocardial cells. At the same time, the purity of myocardial cells can also be improved greatly.

Objective To investigate therapeutic effects of phlorizin on the early stage of diabetic retinopathy(DR)in diabetic db/db mice.Methods The 24 db/db male mice were divided into 3 groups of phlorizin-treated diabetic group(20mg/kg of phlorizin by intragastric gavage)(DMT,n =8),diabetic group(normal saline solution by intragastric gavage)(DM,n=8),with C57BLKS/J db/ m mice as the control group(CC,n=8)for ten weeks.After ten weeks,all mice were sacrificed and the retina was isolated.The specimen was embedded in paraffin wax and sliced.The sections were dyed with Sudan red and observed for retinal morphological change.Meanwhile,the expressions of GSK-3β and phospho-GSK-3β in retinas were determined by western blot.Results During the study,the CC group showed good condition.The DM group featured with polydipsia,polyphagia and a rapid increase of body weight.Phlorizin-treated diabetic mice displayed much better than the DM group.Compared with CC group,DM group showed cells proliferation in capillary endothelia and obvious thickening in basement membrane.After administration of phlorizin,the vascular lesion of central regions was remarkably improved.The expression level of phospho-GSK-3β protein was lower in DM group than in control group(76.2±15.8 vs.255.4 ± 10.7,t =16.30,P＜0.01)and was higher in Phlorizin-treated DM group(188.5±18.4)than in DM group(76.2± 15.8,t=8.05,P＜0.01).Conclusions Phlorizin decreases phospho-GSK-3β3expression in retina in db/db diabetic mice and suppresses the GSK-3βactivity,which leads to the protection of DR.

Objective To establish a proper internal quality control(QC) system for the automated hematology analyzer accord‐ing to the corresponding stipulation by ISO15189 :2012 .Methods The high ,normal and low levels of controls offered by the instru‐ment were used to detect 22 items in whole blood cells routine analysis ,draw the Levey‐Jennings chart ,analyze and evaluate the quality control data for each month or each batch .Results A proper QC system was established ,including quality goals setting , new QC target value and standard deviation calculating and the QC rules choosing .In setting the target value for new batch of con‐trol ,MCV ,HCT and MCHC should adopt the mean values of at least 20 values ,while other items could adopt the mean values of 10 values ;the difference of CV values for some items existed in different QC levels ,different brands or different instruments in the same brand .Therefore ,the quality goals and standard deviation for new batch should be set according to different levels and differ‐ent instruments .Due to the QC characteristics and existence of QC chart drift phenomenon ,therefore MCV ,HCT and MCHC should adopt the 13 s and 22 s rules .For the other items ,the 13 s ,22 s and 10x rules were adoted .Conclusion Under the corresponding stipulations of ISO15189 and by combining with the laboratory practical situation ,a set of applicable internal quality control system is established .

Objective To analyze the regulatory T cell ( CD4 +CD25 +FoxP3 +T) levels in peripheral blood of patients with cavitary tuberculosis before and after surgery .Methods We compared the proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells in 13 patients with cavity tuberculosis pre-and post-operatively and in 10 healthy control subjects with flow cytometry .Results (1) The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells were significantly higher in patients with cavity tuberculosis and at 1-month postoperatively than that in healthy controls(P<0.01).The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells had no statistical significance between the patients with cavity tuberculosis at 6-months postoperatively and healthy controls (P>0.05).(2)The proportion of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells decreased significantly af-ter 6 months surgery than preoperatively in patients with cavity tuberculosis (P<0.01).(3)Pre-and post-operative proportions of CD4 +CD25high T cells and CD4 +CD25 +FoxP3 +T cells showed a positive correlation ( r=0.878,P<0.01 ) .Conclusion The proportion of the circulating T regulatory cells increases in patients with cavity tuberculosis , and decreases after surgery .Infection with M.tuberculosis may induce regulatory T cell-surface molecular changes.

Objective This study was designed to determine the effect of transient receptor potential vanilloid type 4(TRPV4)on angiotensin Ⅱ(Ang Ⅱ)-induced renal injury in TRPV4-null mutant(TRPV4 -/-)mice. Methods The mice were divided into sham group and Ang Ⅱ-treated group. Ang Ⅱ was infused systemically into wild type(WT)and TRPV4 -/- mice via a miniosmotic pump for 4 weeks, and the sham mice were given with normal saline. Systolic blood pressure,urinary excretion of albumin and 8-isoprostane, serum creatinine, and the pathological changes in the kidney tissues were assayed after the 4-week treatment. Results Compared with corresponding sham mice,Ang Ⅱ infusion led to enhanced systolic blood pressure,increased urinary excretion of albumin and 8-isoprostane,increased serum creatinine(P< 0.05),and enhanced glomerulosclerosis degree and renal tubulointerstitial injury index(P< 0.05)in the WT and TRPV4 -/- mice. The result were associated with enhanced collagen levels in the kidney(P< 0.05). All of them were attenuated by the deletion of TRPV4 in the absence of alteration in blood pressure(P< 0.05). Conclusions Deletion of TRPV4 could alleviate renal injury during Ang Ⅱ-induced hypertension, suggesting that TRPV4 may contribute to the pathophysiology of angiotensin Ⅱ-induced renal injury.

Objective: To study the effect of nutrition intervention on the body composition and blood glucose in newly diagnosed patients with type 2 diabetes mellitus complicated with overweight and obesity. Methods: A total of 84 newly diagnosed patients with type 2 diabetes mellitus complicated with overweight and obesity were selected from the department of endocrinology in China-Japan Friendship Hospital from April 2015 to December 2016. Basic information and body composition of these patients were collected and measured. Also the food frequency questionnaire (FFQ) was used to investigate the dietary status in the past month, and calculate the intake of energy, three major nutrients and dietary fiber. All patients received nutrition intervention by nutritionist for three months as requested by Diabetes guidelines. After 3-month intervention, blood glucose, body composition and dietary status were examined again. Relevant indicators of patients were compared before and after the intervention. All patients were divided into three groups according to the degree of body mass index (BMI)′s change: <5%, 5%-10% and >10%. The differences of body weight, muscle, body fat rate, visceral fat index and blood glucose level among three groups were quantified. Results: There were 53 male patients with age (41.5±8.9) years and HbA1c (7.5±0.4)%, and 31 female patients with age (40.1±8.5) years with HbA1c (7.6±0.5)%. The intake of energy, carbohydrate and fat of patients were significantly decreased (P<0.05) and dietary fiber was significantly increased (P<0.05) after nutrition intervention. The body weight, muscle mass, fat mass and visceral fat index were also significantly decreased (P<0.05). The average decrease of fat was about 2.8 kg accounting for 73.7% (2.8 kg/3.8 kg) of the total weight loss. The body fat rate decreased from (31.2±3.1)% to (28.8±3.2)% (P<0.05). The fasting blood glucose and postprandial blood glucose were both decreased significantly after intervention (P<0.05). Among three BMI decreased groups, there was no significant difference in body weight and muscle mass (P>0.05), but a significant difference in body fat rate and visceral fat index (P<0.05). The body fat rate, visceral fat index, fasting blood glucose and postprandial blood glucose had more decreases with the greater amount of weight loss (P<0.05). Conclusion The three-month nutrition intervention could change the dietary habit and components of patients with type 2 diabetes mellitus by reducing the blood glucose, body fat rate and visceral fat index significantly. The degree of reduced BMI is positively related to the decrease of fasting blood glucose and postprandial blood glucose.

Objective:To investigate the expression of estrogen receptor alpha (ERα) protein and BRAF V600E protein in thyroid papillary carcinoma (PTC) and their relationship with clinical factors of PTC. Methods:The expression of ERα and BRAF V600E protein in 1 105 PTC patients was detected by immunohistochemistry. The relationship among ERα, BRAF V600E protein and clinical factors were analyzed. Results:Positive ERα protein was correlated with maleness (χ 2= 6.087, P=0.001), age< 45 years old (χ 2=5.197, P=0.023) and multifocal tumors (χ 2=4.446, P=0.035). Positive BRAF V600E protein was correlated with positive ERα protein (χ 2=6.209, P=0.013), Hashimoto thyroiditis (χ 2=29.388, P<0.001), no lateral lymph node metastasis (χ 2=6.849, P=0.009) and multifocal tumors (χ 2=9.596, P=0.035). Conclusions:ERα expression is more common in male patients, patients younger than 45 years of age, those with multifocal tumors and positive BRAF V600E protein. BRAF V600E protein may inhibit Hashimoto's thyroiditis, tumor growth and the occurrence of lateral lymph node metastasis, and promote the occurrence of multiple focal tumors.

Myocardial fibrosis is featured by excessive proliferation of cardiac fibroblast and collagen deposition. There is close relationship between myocardial fibrosis and various cardiovascular disease, and it is potential risk factor for sudden death. At present the precise mechanism remains unclear.Myocardial fibrosis has been found to relate with renin-angiotensin-aldosterone system,cell factor,oxidative stress,inflamatory factor,endothelial function obstacles,Intracellular calcium ion. These factors influences the occurrence of myocardial fibrosis by the same or different pathway.

Liver cirrhosis-related cardiomyopathy (CCM) is a common and easily overlooked complication associated with liver cirrhosis. Because of the high incidence and impact on success of the liver transplantation and the transcarotid intrahepatic portosystemic shunt surgery, it has attracted the attention of researchers in the field recently. Liver cirrhosis patients have liver morphological alterations, which result in increased serum level of total bile acids and changes in bile acid composition. The increased bile acid concentrations are cardiotoxic and change cardiac functions. This study discussed and summarized recent advancements in the role of bile acids in cirrhosis-related cardiomyopathy in three aspects, i.e., changes in bile acid metabolism in liver cirrhosis patients, effects of bile acid on changes in cardiac functions, and ursodeoxycholic acid as a potential therapeutic agent for CCM. This review expects to provide novel approaches for future prevention and treatment of CCM.

OBJECTIVE： To observe the effects of Shenfu yixin decoction on reactive oxygen species （ROS） and energy metabolism in primary hypoxic cardiomyocytes. METHODS： After isolation， culture and identification， primary cardiomyocytes of neonatal SD rats were randomly divided into normal group， model group， positive control group （coenzyme Q10， 0.1 mmol/L） and Shenfu yixin decoction low-dose and high-dose groups （0.25， 0.5 mg/mL）. Except for normal group， other groups were cultured with 5％O2， 5％CO2 and 90％N2 for 6 h to induce hypoxic injury model. After 6 hours of hypoxia， ROS contents in cardiomyocytes and mitochondria of each group were detected by ROS probe and flow cytometry. Luciferase luminescence and Western blotting were used to detect ATP content and CK protein expression of each group. Transmission electron microscope was used to observe ultrastructure of cardiomyocytes in each group. RESULTS： Compared with normal group， the expression of ROS in primary hypoxic cardiomyocytes and mitochondria as well as the content of ROS were increased significantly， while the content of ATP and expression levels of CK protein were decreased significantly （P＜0.05）； there were swelling of endoplasmic reticulum and mitochondria， dissolution or even disappearance of mitochondrial ridge， obvious cardiomyocytes injury. Compared with model group， the expression of ROS in primary hypoxic cardiomyocytes and mitochondria of administration groups， the contents of ROS in primary hypoxic cardiomyocytes of positive control group and Shenfu yixin decoction high-dose group as well as the content of ROS in primary hypoxic cardiomyocytes mitochondria of administration groups were all decreased significantly， while ATP contents in primary hypoxic cardiomyocytes of positive control group and Shenfu yixin decoction high-dose group as well as expression levels of CK protein in primary hypoxic cardiomyocytes of administration groups were all increased significantly （P＜0.05）. The primary hypoxic cardiomyocytes injury was relieved significantly in positive control group and Shenfu yixin decoction high-dose group. CONCLUSIONS： Shenfu yixin decoction can improve primary hypoxic cardiomyocytes， down-regulate the expression of ROS in cardiomyocytes and mitochondria and also improve its energy metabolism.