Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.

Adult ; Anti-HIV Agents ; therapeutic use ; Familial Primary Pulmonary Hypertension ; HIV ; pathogenicity ; Hepacivirus ; pathogenicity ; Hepatitis B virus ; pathogenicity ; Humans ; Hypertension, Pulmonary ; drug therapy ; virology ; Male ; Sulfonamides ; therapeutic use

BACKGROUNDDiabetic myocardiopathy is characterized by myocardial interstitial fibrosis and cardiac dysfunction. Statins were found to exert protective effects on cardiovascular disease by suppressing activation of small G proteins, independently of their lipid-lowering effect. The study investigated the effect of fluvastatin on myocardial interstitial fibrosis, cardiac function and mechanism of its action in diabetic rats.

METHODSTwenty-four male SD rats were randomly assigned to 3 groups: control rats (n = 8), streptozotocin (STZ)-induced diabetic rats (n = 8), and diabetic rats treated with fluvastatin (administered fluvastatin orally, 10 mg/kg body weight per day, n = 8). Twelve weeks later, miniature cardiac catheter was inserted into the left ventricle to conduct hemodynamic examination. Then myocardium tissues were collected, collagen content was detected by picro-sirius red staining, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of connective tissue growth factor (CTGF), and Western blotting was used to detect the protein expression of CTGF. Rho activity was determined by pull-down assay.

RESULTSAfter 12 weeks, the left ventricular systolic pressure (LVSP) and maximum rate of left ventricular (LV) pressure rise and fall (+dP/dt max and -dP/dt max) were significantly lower and left ventricular end diastolic pressure (LVEDP) was higher in the diabetic rats than those in the control rats (P < 0.01). Moreover, in LV myocardial tissue of diabetic rats the collagen content, fibronectin, mRNA and protein expression of CTGF and the activity of RhoA were all significantly increased compared with the control rats (P < 0.01). Administration of fluvastain obviously improved the cardiac function of diabetic rats, attenuated fibronectin expression, mRNA and protein expression of CTGF and the activity of RhoA in LV myocardium of diabetic rats.

CONCLUSIONSFluvastatin attenuates cardiac dysfunction and myocardial interstitial fibrosis of diabetic rat by inhibiting activity of RhoA to down-regulate the overexpression of CTGF, and Rho/Rho-kinase pathway may be an important target in the treatment of diabetic cardiomyopathy.

Animals ; Anticholesteremic Agents ; therapeutic use ; Blood Glucose ; metabolism ; Blotting, Western ; Cholesterol ; blood ; Connective Tissue Growth Factor ; metabolism ; Fatty Acids, Monounsaturated ; therapeutic use ; Fibrosis ; blood ; drug therapy ; metabolism ; Hemodynamics ; drug effects ; Immunohistochemistry ; Indoles ; therapeutic use ; Male ; Myocardium ; metabolism ; pathology ; Random Allocation ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the clinicopathological features of nonalcoholic steatohepatitis (NASH) and elucidate its diagnosis and differential diagnosis.

METHODSLiver biopsy tissues and clinical data of 32 patients with NASH were collected and the clinicopathological findings by HE and Masson staining were evaluated for NASH grading.

RESULTSBallooning degeneration of the liver cells and fibrosis around hepatic sinusoid was scarce in mild NASH cases and increased in moderate to severe cases. Steatotic and inflammatory cells in the liver lobes decrease in liver cirrhosis related to seatohepatitis.

CONCLUSIONBallooning degeneration of the liver cells and fibrosis around the hepatic sinusoid have important value in differential diagnosis of mild from moderate to severe NASH, and correct histological grading benefits clinical intervention and prognostic evaluation of NASH.

Adult ; Biopsy, Needle ; Diagnosis, Differential ; Fatty Liver ; diagnosis ; pathology ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Liver ; pathology ; Male ; Middle Aged ; Prognosis

OBJECTIVE: To observe the effect of anti-osteoporosis drugs on the curative effect of femoral head replacement in the elderly patients with proximal humerus fracture. METHODS: From November 2012 to June 2016, 38 patients with proximal humeral fractures received humeral head replacement were divided into the treatment group and the control group according to whether the anti-osteoporosis drugs were used after the operation. The treatment group included 19 cases, of which 11 cases were three part fractures, 18 cases were four part fractures, and bone density was(0.58±0.14) g/cm²; the control group involved 19 cases, of which 10 cases were the three part fractures, 9 cases were four part fractures, and bone density was(0.58±0.11) g/cm². Periprosthetic bone mineral density(BMD) was measured at 4, 8, 12, 24 and 48 weeks after operation, and visual analogue scale(VAS) was used to evaluate the pain and Neer score was used to evaluate the function of the shoulder joint. RESULTS: The incisions of all patients were healed with grade A and no complications occurred. Thirty-five patients were followed up for 1 year. The bone density around the prosthesis of treatment group was higher than that of control group, the difference was statistically significant(<0.05);VAS in two groups had no statistical significance(>0.05). The total score and functional score of Neer in the treatment group were better than those in the control group, the difference was statistically significant(<0.05), and there was no significant difference in pain and activity score between the two groups(>0.05). According to the Neer score, the results of treatment group was excellent in 10 cases, good in 5 cases, fair in 3 cases;in the control group, 3 cases were excellent, 9 cases were good, and 5 cases were fair;the difference between the two groups was statistically significant(<0.05). CONCLUSIONS Artificial humeral head replacement combined with anti-osteoporosis drugs in the treatment of proximal humeral fractures in elderly patients can effectively improve the bone density around the prosthesis and restore shoulder function. The early clinical effect is satisfactory.
Aged ; Calcitonin ; therapeutic use ; Fracture Fixation, Internal ; Humans ; Humeral Head ; Postoperative Period ; Shoulder Fractures ; prevention & control ; Shoulder Joint ; Treatment Outcome

Our previous data demonstrate that impairment of arterial baroreceptor reflex (ABR) plays an independent role in hypertension target organ damage. To elucidate the mechanisms responsible for the dysfunction of ABR associated organ damage, sinoaortic denervated (SAD) rats were used as an animal model of ABR dysfunction. Twenty-four-hour continuous blood pressure (SBP and DBP), blood pressure variability (BPV), heart rate (HR) and HR variability (HRV) were measured in conscious and unrestrained rats. Angiotensin II (Ang II) in plasma, heart and kidney was assayed by raio-immunological assay (RIA) 1 or 18 weeks after denervation. In short-term SAD rats, twenty-four-hour mean SBP and DBP increased compared with that of sham-operated rats and long-term SAD rats. No significant difference in SBP, DBP or HR was found between long-term SAD rats and sham-operated ones. Compared with the sham-operated rats, long-term SAD rats had elevated BPV. No significant change in Ang II levels of caridiac and renal tissues was found in short-term SAD rats. In long-term SAD rats, Ang II level of plasma was not increased while the Ang II content in the heart and kidney increased. Ang II contents of plasma and tissues in long-term SAD rats exposed to chronic stress were higher than those in the control rats. These results show (1) in short-term SAD rats blood pressure increased, while in long-term SAD rats 24 h mean blood pressure did not increase, although BPV elevated in long-term SAD rats; (2) in long-term SAD rats, secretion of Ang II in cardiac and renal tissues was enhanced and more Ang II released when the animals were exposed to chronic stress. These results suggest that elevated BPV and secretion of Ang II may be related to the development of organ damage induced by ABR dysfunction.
Angiotensin II ; blood ; metabolism ; Animals ; Aorta ; innervation ; metabolism ; Autonomic Denervation ; Baroreflex ; physiology ; Blood Pressure ; physiology ; Carotid Sinus ; innervation ; metabolism ; Hypertension ; metabolism ; physiopathology ; Kidney ; metabolism ; Male ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley

Hierarchical diagnosis and treatment is the top priority of health care reform,it will provide a solid institutional foundation and institutional protection for the construction of healthy China and the basic health care system.The paper summarizes the present practice of the hierarchical diagnosis and treatment in China,and analyzes the existing problems,on this basis puts forward corresponding countermeasures and suggestions as the references for the construction of hierarchical diagnosis and treatment and deepening reform.

ObjectiveTo investigate the development trend of drinking water type of endemic fluorosis in Qinghai province, and to provide the basis for the prevention and treatment of the disease. MethodsIn 2009, six monitoring counties were chosen by using simple random sampling methods, all diseased villages of the six monitoring counties were classified into light, moderate and severe disease types according to water fluorine content on the historical data, and 1 village was respectively chosen from each type. In monitoring villages with improved water, 3 tap water and one source water samples were collected, respectively. Five water samples were collected randomly in water unimproved monitoring villages according to water well locations of east, west, south, north and center. The fluorine content in water and urine was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006). Children aged 8 to 12 were examined for dental fluorosis by Dean method.Clinical osteofluorosis of all the resident over the age of 16 was examined, 2 village of these counties were randomly selected, and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray using Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008). Urine sample of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method (WS/T 89-2006). ResultsImproving water projects had been implemented in 14 monitoring villages of the 18 villages in 6 counties, the rate of improved-water was 77.78％(14/18). Among the 14 projects, 5 improved-water projects ran normally, and 9 projects ran with intermittently water supply. Seventy-five water samples were tested, themean of water fluoride was 0.48 mg/L. The prevalence of dental fluorosis was 31.95％ (285/892), that of clinical skeletal fluorosis was 36.55％(1570/4295) and the X-ray detection rate of skeletal fluorosis was 25.64％ (20/78).Five hundred and seventy-one urine samples of children were determined, and geometric mean of urinary fluorine was 1.04 mg/L; 370 adult urine samples were determined, and geometric mean of urinary fluorine was 1.52 mg/L Conclusion Epidemic of drinking water type of endemic fluorosis is still serious in Qinghai province, and drinking water defluoride measures should be further strengthened and improved.

Objective To provide anatomic evidence for chock-block technique used for acromioplasty.Methods Thirty cadaveric specimens of intact shoulder fixed by formaldehyde solution were used for this anatomic study.They were chosen from 15 males and 15 females and from 17 left and 13 right sides.First the specimens were dissected for observation of general morphology of the acromion.According to the Bigliani's classification,the acromions were sorted in morphology as type Ⅰ (flat),type Ⅱ (curved) and type Ⅲ (hooked).The shortest distance from the acromion to the humeral head (A-H) was measured.Then the location of subacromial impingement was observed and the thickness of the anterolateral angle was measured preoperatively.At last,acromioplasty using the chock-block technique was performed in the acromions of types Ⅱ and Ⅲ.Postoperatively,the distance A-H and the thickness of the anterolateral angle were measured again for comparison with the preoperative values.Results According to the Bigliani's classification,7 acromions were type Ⅰ,13 type Ⅱ and 10 type Ⅲ.Subacromial impact occurred only in the anterior acromions of types Ⅱ and Ⅲ.The distance A-H was 0.670 ± 0.035 cm in type Ⅰ acromions,significantly longer than the preoperative values of type Ⅱ (0.506 ±0.029 cm) and type Ⅲ (0.334 ±0.023 cm) (P ＜ 0.05),but showed no significant differences from the postoperative values of type Ⅱ (0.692 ±0.025 cm) and type Ⅲ (0.699 ±0.024 cm) (P ＞ 0.05).The anterolateral angular thickness of type Ⅰ acromions was O.413 ± 0.015 cm,significantly thinner than the preoperative values of type Ⅱ (0.607 ±0.014 cm) and type Ⅲ (0.623 ±0.025 cm) (P ＜ 0.05),but showed no significant differences from the postoperative values of type Ⅱ (0.414±0.035 cm) and type Ⅲ (0.412±0.033 cm) (P＞ 0.05).Conclusion The chock-block technique used for acromioplasty is effective because it can enlarge the supraspinatus outlet and deter impingement.

OBJECTIVETo investigate the significance of Rac subfamily members in the gastrointestinal carcinogenesis and progression.

METHODSThe mRNA expression of Rac1, Rac2 and Rac3 in 12 kinds of gastrointestinal cancer cell lines was examined by semi-quantitative RT-PCR. The activities of Rac1 protein in 5 kinds of gastric cancer cell lines were tested by pull-down assay.

RESULTSCompared with the normal gastric mucosa and intestinal epithelial cell line, the mRNA expression of Rac1 and Rac3 was up-regulated in most of gastrointestinal cancer cell lines. The activities of Rac1 protein increased markedly in gastric cancer cell lines.

CONCLUSIONThe increased mRNA expression of Rac1 and Rac3 in gastrointestinal cancer cell lines and the abnormal activation of Rac1 protein in gastric cancer cell lines might be correlated with the carcinogenesis of gastrointestinal cancer.

Cell Line, Tumor ; Gastrointestinal Neoplasms ; metabolism ; Humans ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; rac GTP-Binding Proteins ; genetics ; rac1 GTP-Binding Protein ; analysis ; genetics