BACKGROUND: Proliferation and differentiation of mesenchymal stem cells (MSCs) is associated with platelet concentration in platelet-rich plasma (PRP). Low enrich multiple cannot reach proper effects, but high level had inhibitory effects on osteoanagenesis.OBJECTIVE: To observe the effect of different volume fraction of PRP on dog BMSC proliferation.DESIGN: A cytological in vitro study.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: Dog BMSCs of 5 passage were adjusted to 3×10~8/L, and incubated in a 96-well plate at 200 μL per well. Following 24 hours of routine culture, primary medium and non-adherent cells were discarded. Prepared PRP gel was mixed with serum-free low-glucose DMEM containing penicillin and streptomycin, and then diluted into 5%, 6.25%, 7.5%, 8.75%, 10% volume fraction. 200 μL above-described liquid was added into the 96-well plate, which was subsequently placed in a incubator.We set up a blank control.MAIN OUTCOME MEASURES: MTT was used to investigate effect of different volume fraction of PRP on dog BMSC proliferation.RESULTS: Compared with the blank control group, various volume fraction of PLP could promote dog BMSC proliferation in early stage. With prolonged time, proliferation speed began to increase at day 6 in the 8.75% and 10% PRP groups, entering platform stage. BMSC number was increased rapidly in the 5% and 6.25% PRP groups, especially in the 6.25% PRP group.CONCLUSION: PRP gel could promote BMSC proliferation markedly and proliferation strength of BMSCs was correlated to the density of PRP. BMSC proliferation would be accelerated by the low density of PRP.

BACKGROUND: Platelet-rich plasma (PRP) contains abundant growth factors that were needed for osteanagenesis. Moreover,the proportion of each growth factor formed by an organism, with good synergism.OBJECTIVE: To explore the influence of PRP on osteogenetic activity of canine bone marrow mesenchymal stem cells (BMSCs) after induction in vitro.DESIGN, TIME AND SETTING: The in vitro cytological experiment was performed at the Central Laboratory of Xiangya Hospital of Central South University from June 2007 to February 2008.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: The 3~(rd) generation BMSCs were collected and divided into 4 groups. BMSCs in the control group were incubated in standard medium. BMSCs in the osteogenetic induction medium group were incubated in high-glucose DMEM containing fetal calf serum, dexamethasone, beta-sodium glycerophosphate and vitamin C. BMSCs in the PRP group were incubated in low-glucose DMEM containing 6.25% PRP. BMSCs in the combination group were incubated in high-glucose DMEM containing 6.25% PRP, dexamethasone, beta-sodium glycerophosphate, and vitamin C.MAIN OUTCOME MEASURES: Alkaline phosphatase activities were measured. Expression of collagen type I was examined by immunocytochemical staining. Calcium tuberoses were labeled using modified Von Kossa staining. Expression of osteocalcin mRNA was examined by RT-PCR.RESULTS: Levels of alkaline phosphates of all groups became increased along with time. The alkaline phosphates level of combination group was strongest (P < 0.05). Following 7 and 14 days of induction, type I collagen expressed positively in the osteogenetic induction medium and combination groups, but negatively in the PRP and control groups. Following 14 days,formation of calcium nodules were found in the osteogenetic induction medium and combination groups. Following 7 and 14 days,expression of osteocalcin mRNA were similar between the control and PRP groups (P > 0.05), which was significantly lower than the osteogenetic induction medium and combination groups (P < 0.05). Expression of osteocalcin mRNA was significantly lower in the osteogenetic induction medium group compared with the combination group (P < 0.05).CONCLUSION: PRP gel can effectively promote osteoblastic effect of BMSCs after induction in vitro following induction in osteogenetic medium.

Objective To investigate role of N-(30,40-dimethoxycinnamonyl) anthranilic acid (3,4-DAA) in mediating the negative immune regulation by indoleamine 2,3-dioxygenase in mice skin transplantation.Method The mice skin transplant model was established.T lymphocytes were isolated from mice splenocytes,and serum was collected.ELISA was applied to detect the IL-2 and IFN-γ concentrations in culture supernates of mice spleen lymphocytes and serum.Flow cytometry was applied to examine the phenotypes of T lymphocytes and RT-PCR to analyze the IDO mRNA transcription.Results T lymphocyte proliferation was significantly decreased by 3,4-DAA and the concentrations of IL-2 and IFN-γ were apparently decreased in 3,4-DAA group as compared with those in control group.The percentages of CD3 +,CD4 + and CD8 + T lymphocytes were lower,and the percentage of CD4 + CD25 + T lymphocytes was higher in 3,4-DAA disposal group than in control group,but there was no significant difference.IDO mRNA expression showed obvious increase in 3,4-DAA group as compared with control group.Conclusion 3,4-DAA can up-regulate the IDO expression to induce the increasing metabolism of tryptophan,subsequently inhibiting T cell proliferation and showing specific property of negative immune regulation.

The purpose of this study was to assess the factors affecting caudal screw loosening after spinopelvic fixation for adult patients with spinal deformity. This meta-analysis calculated the weighted mean difference (WMD) and odds ratio (OR) using Review Manager ver. 5.3 (RevMan; Cochrane, London, UK). The loosening group was older than the control group (WMD, 2.17; 95% confidence interval [CI], 0.48–3.87; p=0.01). The S2 alar-iliac (S2AI) could prevent the caudal screw from loosening (OR, 0.43; 95% CI, 0.20–0.94; p=0.03). However, gender distribution (p=0.36), the number of fusion segments (p=0.24), rod breakage (p=0.97), T-score (p=0.10), and proximal junctional kyphosis (p=0.75) demonstrated no difference. Preoperatively, only pelvic incidence (PI) in the loosening group was higher (WMD, 5.08; 95% CI, 2.71–7.45; p<0.01), while thoracic kyphosis (p=0.09), lumbar lordosis (LL) (p=0.69), pelvic tilt (PT) (p=0.31), pelvic incidence minus lumbar lordosis (PI–LL) (p=0.35), sagittal vertical axis (SVA) (p=0.27), and T1 pelvic angle (TPA) demonstrated no difference (p=0.10). PI–LL (WMD, 6.05; 95% CI, 0.96–11.14; p=0.02), PT (WMD, 4.12; 95% CI, 0.99–7.26; p=0.01), TPA (WMD, 4.72; 95% CI, 2.35–7.09; p<0.01), and SVA (WMD, 13.35; 95% CI, 2.83–3.87; p=0.001) were higher in the screw loosening group immediately postoperatively. However, TK (p=0.24) and LL (p=0.44) demonstrated no difference. TPA (WMD, 8.38; 95% CI, 3.30–13.47; p<0.01), PT (WMD, 6.01; 95% CI, 1.47–10.55; p=0.01), and SVA (WMD, 23.13; 95% CI, 12.06–34.21; p<0.01) were higher in the screw loosening group at the final follow-up. However, PI–LL (p=0.17) demonstrated no significant difference. Elderly individuals were more susceptible to the caudal screw loosening, and the S2AI screw might better reduce the caudal screw loosening rate than the iliac screws. The lumbar lordosis and sagittal alignment should be reconstructed properly to prevent the caudal screw from loosening. Measures to block sagittal alignment deterioration could also prevent the caudal screw from loosening.

Objective To analyze the outcome of surveillance results on plague and to provide the evidences for the policy making in Longlin county Guangxi. Methods The epidemic data and the surveillance results of plague were analyzed and assessed with epidemiology methods in Longlin county Guangxi from 2000 to 2009, and the density of rodents, the rodents infected with flea, flea index and other indicators were calculated. Regional composition of the rats and fleas were analyzed. Results A totally of 4829 rats were captured and 4737 fleas were collected in the past 10 years, Rattus Flavipestus(81.92%,3956/4829) and Xenopsylla Cheopis (79.04%,3744/4737) were dominant species. The annual average density of rodents, the rodents infected with flea, index of flea were 3.30%(4829/146 206), 27.99%(1351/4827) and 0.98(4737/4827), respectively. A totally of 4792 rats were examined and 10 strains Yersinia Pestis were isolated. Indirect hemorrhagic assessed(IHA) was used to test the F1 antibody against plague in the blood serum of the rats and indicator animals, and 3 positive rats and 24 positive animals were found, respectively. Twenty seven natural villages in 3 towns had been involved in the plague. Conclusions The plague foci exists in Longlin county of Guangxi province. The plague foci in the areas have the same feature with the plague foci of Rattus Flavipectus. There is a potential risk for plague in this region, we should improve the quality of surveillance, increase indicator animals of the plague, and try to apply new surveillance method.

Objective To learn the epidemiological characteristics of foodborne disease outbreak in 2015,Zhejiang Province,and to provide evidences for the prevention and control of foodborne disease outbreak.Methods Descriptive epidemiological analysis using ArcGIS 10.2 and SPSS 19.0 was conducted on geographical country data,information of permanent resident population and the incidence data of foodborne disease outbreak reported through foodborne disease outbreak reporting information system in 2015.Results There were 116 foodborne disease events reported in Zhejiang in 2015,resulting in 1 009 persons attacked,115 hospitalization and 10 deaths.The proportion of unknown pathogenic factor for events was 29.31%.The events appearance sporadic,there were some local hotspots in Jiaxing.The vibrio parahaemolyticus was the most common pathogen (32.76%),and poisonous mushrooms(8.62%)come the second.Most foodborne disease incidents occurred during June to November.Restaurants ,family and rural feasts were the major places where the foodborne disease occurred.The proportion of family foodborne disease events because of mistaken and misuse poisonous substances increased,and the hospitalization rate and mortality of events of which were the highest (33.9% and 7.63% respectively).Conclusion The food safety management and of restaurants and rural feasts should be strengthened in summer and autumn,as well as foodborne disease prediction and forewarning to reduce the foodborne disease incidents caused by microorganism contamination and poisonous muchnooms.It is necessary to conduct public risk cognition research for foodborne disease in Zhejiang,which could provide evidence for preventing and controlling family foodborne disease.

BACKGROUNDA new inhalable insulin aerosol (Inh-Ins) was developed in China. The aim of this multicenter clinical study was to evaluate the efficacy and safety of this new Inh-Ins as a treatment of type 2 diabetes. Regular porcine insulin (RI) was used as a control.

METHODSThis study is a prospective, randomized, open-label, parallel-group multicenter clinical trial in which 253 qualified patients with type 2 diabetes received the insulin Glargine daily at bedtime plus either a pre-meal Inh-Ins or a pre-meal subcutaneous RI for 12 weeks. HbA1c, fasting plasma glucose (FPG), the 1-hour-postprandial blood glucose (1hPBG) and the 2-hour-postprandial blood glucose (2hPBG) were measured. Events were monitored for adverse effects.

RESULTSAfter 12 weeks, the HbA1c decreased significantly from baseline in both treatment groups, with no significant difference between the two regimens. In the Inh-Ins group, FPG, both 1hPBG and 2hPBG significantly declined from baseline after the 8th- and 12th-weeks of treatment. The reduced values of FPG or 1hPBG between the two groups showed a more significant hypoglycemic effect with the Inh-Ins than the RI. After 12 weeks, the pulmonary carbon monoxide diffusing capacity (DLco) was significantly lower in Inh-Ins group than in the RI. The main side effects of Inh-Ins were coughing, excessive sputum, and hypoglycemia.

CONCLUSIONSInh-Ins was effective in decreasing HbA1c like the RI. It was better in lowering the FPG and the 1hPBG than the RI. Its main side effects were coughing, excessive sputum, and hypoglycemia. Also, Inh-Ins slightly impaired DLco.

Adolescent ; Adult ; Aerosols ; Aged ; Blood Glucose ; analysis ; Body Weight ; drug effects ; Cough ; chemically induced ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Female ; Glycated Hemoglobin A ; analysis ; Humans ; Hypoglycemia ; chemically induced ; Insulin ; administration & dosage ; adverse effects ; Male ; Middle Aged ; Prospective Studies

OBJECTIVETo explore the role of the autoantibodies against M(2)-muscarinic receptor (M(2)-receptor), beta(1)-adrenergic receptor (beta(1)-receptor) in the development of diabetic with refractory hypertension.

METHODSSerum autoantibodies against M(2) and beta(1) were detected by ELISA using synthesized epitopes of the second extracellular loop of M(2) receptor (169 - 193) and beta(1) receptor (197 - 222) in healthy controls (n = 40), diabetic patients (n = 62), diabetic patients with non-refractory hypertension (n = 55) and diabetic patients with refractory hypertension (n = 81).

RESULTSThe positive rates of the autoantibodies against M(2) receptor and beta(1) receptor were similar among healthy controls (15.0% and 17. 5%), diabetes mellitus patients (17.7% and 14.5%) and diabetic patients with non-refractory hypertension (16.4% and 12.7%) but are significantly higher in diabetic patients with refractory hypertension (64.2% and 55.6%, P < 0.01 vs. other 3 groups).

CONCLUSIONThis finding suggests that autoimmune mechanisms might play a role in the pathogenesis of diabetic patients with refractory hypertension.

Adult ; Autoantibodies ; blood ; Diabetes Mellitus, Type 2 ; blood ; complications ; Female ; Humans ; Hypertension ; blood ; complications ; Male ; Middle Aged ; Receptor, Muscarinic M2 ; immunology ; Receptors, Adrenergic, beta-1 ; immunology

Objective To explore the relation between the positive rates of autoantibodies against β1 adrenergic receptor(β1-receptor) and (M2-receptor) with urinary albumin excretion rate (UAER) in type 2 diabetes patients with refractory hypertension.Methods Autoantibodies against β1-and M2-receptor as well as autoantibodies were determined in type 2 diabetes patients with(n=136)or without(n=11 1) refractory hypertension,hypertensive patients without renal failure (n=60) and healthy contrel snbjects (n=40.control) by ELISA.Resuits The positive rates of the autoantibodies against β1.receptors (44.9%) and M2-receptor(37.5%)in patients with type 2 diabetes with refractory hypertension were significantly higher than those in patients with type 2 diabetes without refractory hypertension (27.9% and 24.3%,respectively.all P<0.05),in patients with hypertension without renal failure(11.7%and 15.0%.all P<0.01) and in healthy controls(8.3%and 7.5%,all P<0.01).In type 2 diabetes patients with refractory hypertension and renal failure (UAER≥200 μg/min),the positive rates of the autoantibodies against β1-receptor(87.1%,27/31)and against M2-receptor (67.7%,21/31) were significantly higher than those in type 2 diabetes patients with refractory hypertension but without renal failure (UAER 20-199 μg/min,46.7%,28/60 and 41.7%,25/60.respectively.all P<0.05).Conclusion The serum β1-and M2-receptor autoantibodies are positively associated with the UAER level and suggest that these autoantibodies against β1 and M2-receptor may play important roles in the pathogenesis of the type 2 diabetes with refractory hypertension.

Objective To observe the positive rates of autoantibodies against β1 adrenergic receptors(β1-receptor)and angiotensin Ⅱ type 1 receptors(AT1-receptor)in type 2 diabetes patients with or without hypertension.Methods The epitopes of the second extracellular loop of β1-receptor(197-222)and AT_1 receptor(165-191)were synthesized and serum autoantibodies were determined in type 2 diabetes patients with hypertension(n=171)or without hypertension(n=106).Left ventricular dimansion was determined by echocardiography.The 24-hour urinary protein was measured by ELISA.The risk factors for enlarged left ventricle were analyzed by multiple logistic regressions.Results The positive rates of the autoantibodies against β1-receptors(45.0%)and AT_1-receptor(46.2%)in patients with type 2 diabetes with hypertension were significantly higher than those in patients with type 2 diabetes without hypertension (16.0%and 10.4%,respectively,all P＜0.01).In type 2 diabetes patients with hypertension and enlarged left ventricle,the positive rates of the autoantibodies against β1-receptor 61.4%(35/57)and against AT_1-receptor 64.9%(37/57)were significantly higher than those in type 2 diabetes patients with normal left ventricular dimension(36.8%,42/114 and 36.8%,42/114,respectively,all P＜0.01).Regression analysis demonstrated that course of disease,systolic pressure,serum autoantibodies against β1 adrenergic receptor and angiotensin Ⅱ type 1 receptors seraautoantibodies were independent risk factors for Jeft ventricular enlargment(all P＜0.05).Conclusion The scram β1 and AT_1-receptor autoantibodies are related to enlarged left ventricle in type 2 diabetes patients with hypertension and suggest that autoantibodies against β1 and AT_1-receptor might play important roles in the pathogenesis of type 2 diabetes patients with hypertension and enlarged left ventricle.