The objective of this study was to develop research of cardiac cells to reestablish 3D tissue architecture in vitro, we performed studies using collagen membrane as three-dimensional scaffold for cardiac cells culture with the principles and methods of tissue engineering. The polymer scaffold provides a 3-D substrate for cell attachment and tissue formation. Cardiac cells isolated by enzymatic digestion from 1d old neonatal rats were seeded to three-dimensional collagen scaffolds and tissue culture plates. The morphology, beating rate and the metabolic indexes, including specific consumption rate of glucose (q(glu)) , specific production rate of lactate (q(lac)), lactate transform rate ( Y(lac/glu)), specific creatine kinase (CK) and lactate dehydrogenase (LDH) activities of cardiac cells cultured on three-dimensional collagen membrane and tissue culture plates were compared. It was found that cells shape and cells' CK and LDH activity was no differences between 3D and 2D cultures and cell beat rate on cell culture cluster was slower than those cells cultured on collagen membrane, However the cell glucose consumption and lactate yield rate of cells cultured on cluster was higher than those cells cultured on collagen membrane. After 5 days of cultivation, cardiac cells cultured on collagen membrane scaffolds organized into three-dimensional (3D) aggregates as opposed to the two-dimensional (2D) aggregates mosaic pattern seen in tissue culture plates, and spontaneous and rhythmical contractile 3D cultures in unison were visible to the naked eye and the area of synchronous contract three-dimensional (3D) aggregates reaches 80cm2. The mean value of q(glu), q(lac) and Y(lac/glu) of cultured on three-dimensional collagen scaffold was 7.37 micromol/10(6) cells/d, 2.92 micromol/10(6) cells/ d and 0.38 micromol/micromol, versus 7.59 micromol/10(6)cells/d, 3.83 micromol/10(6) cells/d and 0.51 micromol/micromol in tissue culture plates. These results demonstrate that cardiac cells immobilized on collagen membrane in 3D cultures maintain similar metabolic activity and contractile function when compared with native cardiac cells. The above results support the idea that engineered cardiac tissue can be used as a model of native tissue for studies of tissue development and function in vitro and eventually for tissue repair in vivo.
Animals ; Biocompatible Materials ; chemistry ; Cells, Cultured ; Collagen ; chemistry ; Flow Cytometry ; Immunohistochemistry ; Microscopy, Electron, Scanning ; Myocytes, Cardiac ; cytology ; ultrastructure ; Rats ; Rats, Wistar ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

OBJECTIVETo study the inhibitory effects of insulin on nuclear factor-kappa B (NF-kappaB) nuclear translocation of vascular endothelial cells induced by burn serum and its correlative mechanism.

METHODSHuman umbilical vein endothelial cells (HUVECs) were cultured in vitro and divided into 5 groups: blank control group (BC, ordinary culture without any stimulation), normal serum control group (NS, cultured with nutrient solution containing 20% healthy human serum), burn serum stimulation group (BS, cultured with nutrient solution containing 20% burn human serum), burn serum+insulin treatment group (BI, cultured with nutrient solution containing 20% burn human serum and 1x10(-7) mol/L insulin), inhibitor pretreatment group [IP, pretreated with 50 micromol/L protein kinase B (Akt) specific inhibitor LY-294002, then cultured with the same medium as used in BI group 30 minutes later] according to the random number table. Six hours later, the injury and apoptosis of HUVECs was respectively observed by the scanning electron microscope and determined by the flow cytometry. Meanwhile, the phosphorylation of inhibitor kappa B-alpha (p-IkappaB-alpha) and Akt (p-Akt) in cytoplasm, and the content of NF-kappaB-p65 in nucleus were determined with Western blot.

RESULTS(1) Compared with those in BC group, HUVECs in BS group shrank obviously with irregular nuclear structure, and intercellular links jagged or vanished. Slight change was observed in HUVECs structure in NS and BI groups, with the cell ductility and nuclear structure much better than those in BS group. (2) The apoptosis rates of HUVECs in BS group [(28.5+/-2.3)%], BI group [(22.3+/-1.8)%], and IP group [(29.7+/-2.4)%] were all obviously higher than that in BC group [(15.7+/-2.2)%, F=14.288, P<0.05 or P<0.01]. There was no significant statistical difference between NS group [(17.0+/-2.5)%] and BC group in apoptosis rate (F=14.288, P>0.05). The apoptosis rate of HUVECs in BI group was obviously lower than that in BS group (F=14.288, P<0.05). (3) Compared with those in BC group, the protein expressions of p-IkappaB-alpha in cytoplasm and NF-kappaB-p65 in nucleus were up-regulated, and the protein expression of p-Akt in cytoplasm was down-regulated in BS and IP groups. The expression levels of the three proteins in NS and BI groups were close to those in BC group.

CONCLUSIONSInsulin could inhibit the IkappaB phosphorylation, and then restrict NF-kappaB nuclear translocation and improve the vascular endothelial cells function accordingly through regulating phosphatidylinositol 3 kinase/Akt pathway.

Apoptosis ; Burns ; blood ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Humans ; I-kappa B Proteins ; metabolism ; Insulin ; pharmacology ; NF-kappa B ; metabolism ; Phosphorylation ; Serum ; metabolism ; Umbilical Veins ; cytology

Objective Few studies are reported on the relationship of subclinical hypothyroidism ( SCHT ) with visfatin and endothelin .This study aimed to investigate their relationship in patients with cerebral infarction . Methods A total of 200 cerebral in-farction patients treated in The Second Affiliated Hospital of Hainan Medical University from January 2011 to July 2017 were divided in-to a control ( with normal thyroid function , n=40 ) , a mild SCHT ( with thyroid-stimulating hormone TSH <10 mIU/L, normal free triiodothyronine FT3 and normal free thyroxine FT4, n=60), a se-vere SCHT (with TSH≥10 mIU/L, normal FT3 and normal FT4, n=60) , and a clinical hypothyroidism ( CHT ) group ( with TSH ≥4 mIU/L, decreased FT3 and decreased FT4, n=40).The mild SCHT patients were subdivided into medication group A and non-medication group A, and the severe SCHT patients into medication group B and non-medication group B, 30 in each group, those in the medica-tion groups A and B treated by routine therapy plus oral levothyrocine , and those in the non-medication groups A and B by routine ther-apy only.We recorded the age, gender, body mass index, blood pressure, blood glucose, blood lipid, and thyroid function of the pa-tients, and compared the levels of visfatin and endothelin among different groups . Results The level of visfatin was significantly ele-vated in the severe SCHT and CHT groups as compared with the controls ([46.3±10.1] and [49.5±13.6] vs [40.2±9.7] ng/mL, P<0.05), and so was it in medication group B as compared with non-medication group B at 6 months after treatment ([42.9±6.4] vs [39.3±5.5] ng/mL, P<0.05). Conclusion Visfatin is closely related with thyroid hormone in cerebral infarction patients with se -vere subclinical hypothyroidism .Examination of thyroid function is necessary for cerebral infarction patients for the sake of early detec -tion of severe subclinical hypothyroidism and timely intervention .

OBJECTIVETo investigate the anti-apoptosis effect of intensive insulin treatment on cardiac myocytes and its underlying mechanism in severe scald rats.

METHODSTwelve SD rats were suffered from 30% TBSA full thickness scald, and they were divided into: IT group [with intravenous injection of isotonic saline including insulin (15 mU x kg(-1) x min(-1)) and 100 g/L glucose], B group [with treatment of isotonic saline (2 mL x kg(-1) x %TBSA(-1) x 8 h(-1)]. Six SD rats received sham burn as controls[sham(S)group, with treatment of fluid at physiologic dose]. + dp/ dtmax (the rate of the rise of left ventricular pressure) and -dp/ dtmax (the rate of the fall of left ventricular pressure)at 6 post burn hour (PBH)were recorded. Apoptosis were determined by TUNEL staining and DNA ladder. The phosphorylation f Akt and protein expression of Bcl-2 in cardiomyocyte were assayed by Western blotting.

RESULTSThe + dp/ dtmax in the S group, IT group and B group at6 PBH were respectively (5.5 +/- 0.5) x 10(3) mm Hg/s, (3.4 +/- 0.4) x 10(3 mm Hg/s and (2.5 +/- 0.5) x 10(3) mm Hg/s (1 mm Hg = 0.133 kPa), the - dp/ dtmax were respectively (4.55 +/- 0.34) x 10(3) mmHg/s, (2.94 +/- 0.22) x 10(3) mm Hg/s and (2.05 +/- 0.19) x 10(3) mmHg/s.The +/- dp/dtmax in IT group was significantly higher than those in B group( P < 0.01). The apoptosis index in B group was (13.1 +/- 3.4)%, which was obviously higher than that in IT group (6.7 +/- 1.8)% and S group (0.6 +/- 0.4)% (P < 0.01). DNA ladder showed that no DNA fragmentation in S group, but obvious DNA fragmentation forming ladder pattern in B group, and no obvious ladder pattern in IT group. The phosphorylation of Akt and level of Bcl-2 protein in B group were markedly higher than those in IT group ( P < 0.05 or P < 0.01).

CONCLUSIONIntensive insulin treatment can upregulate the activity of Akt and enhance the expression of Bcl-2, and they might constitute the mechanisms for anti-apoptosis in cardiomyocyte and protection of cardiac function.

Animals ; Apoptosis ; drug effects ; Burns ; drug therapy ; pathology ; Insulin ; administration & dosage ; pharmacology ; Male ; Myocytes, Cardiac ; cytology ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the protective effect of intensive insulin treatment on the myocardium of severely scalded rats, and to primarily explore its mechanism.

METHODSEighteen SD rats were divided into three groups, with 6 rats in each group. The rats in burn and intensive insulin group were inflicted with 30% TBSA full-thickness injury on the back. Isotonic saline containing 0.12 U/ml insulin solution, and 100 g/L glucose solution were infused into the rats in the intensive insulin group to keep plasma glucose at the level of 4.0 - 6.6 mmol/L (the total fluid amount was 2 ml x kg(-1) x 8h(-1)). In sham burn group,fluid was given according to physiological demand. The same amount of isotonic saline was infused into the rats in burn group. The venous blood was obtained for the detection of plasma glucose contents, and the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP) were recorded via aortic ventricle cannula before scald and at 1, 2, 3, 4, 5, 6 post-scald hours (PSH). The tissue of the left ventricle was harvested at 6 PSH for the detection of troponin T expression in myocardiocytes.

RESULTSPlasma glucose level was increased to (7.6 +/- 1.7) mmol/L - (8.4 +/- 4.7) mmol/L in burn group during 1-6 PSH, which was significantly higher than that in intensive insulin group (4.5 +/- 0.9) mmol/L - (5.2 +/- 1.3) mmol/L, P < 0.01). Compared with the intensive insulin group, LVSP was markedly decreased in the burn group (60 +/- 11 mm Hg vs 72 +/- 8 mm Hg, P < 0.05) at 1 PSH,whereas LVEDP was increased significantly (21.3 +/- 11.3 mmHg vs 11.7 +/- 5.2 mmHg, P < 0.05). Intensive insulin treatment could significantly inhibit the loss of troponin T protein in myofilaments of myocardium.

CONCLUSIONIntensive insulin treatment possesses a protective effect on myocardia function after severe burns, and it may be related to its preventive effect on the loss of contractile protein in cardiocytes.

Animals ; Blood Glucose ; metabolism ; Burns ; drug therapy ; metabolism ; Insulin ; administration & dosage ; therapeutic use ; Male ; Myocardial Contraction ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Troponin T ; metabolism

OBJECTIVETo evaluate the clinical efficacy of a new formulation of albendazole emulsion (AbzE) in cases of liver cystic hydatidosis.

METHODSTwo regimens of AbzE (10 mg.kg(-1).d(-1) and 12.5 mg.kg(-1).d(-1)) were given to 212 patients with liver cystic hydatidosis in courses ranging from 3 months to more than one year. Assessment of drug efficacy was essentially based on imaging signs with ultrasonography as the main tool. Assessments were performed at the end of different courses and in the follow-up study of 1 - 4 years after the cessation of therapy.

RESULTSAt the end of therapeutic courses, the overall cure rate of the 212 cases was 74.5%, with a 99.1% effective rate. In the follow-up study, the cure rate was 83.1%, effective rate was 89.3%, ineffective rate was 0.6%, and recurrence rate was 10.2%. The highest cure rate was observed in cases receiving AbzE 12.5 mg.kg(-1).d(-1) for 9 months. Retreatment of recurrent cases with AbzE obtained satisfactory results.

CONCLUSIONSAbzE surpassed other currently used antihydatidosis drugs or formulations with its promising efficacy and mild side effects, and could be recommended as a drug of choice in the treatment of cystic hydatidosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Albendazole ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; Echinococcosis, Hepatic ; drug therapy ; Emulsions ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the roles of the residential environment and eating habits in the pathogenesis of bronchial asthma in school children.

METHODSOne hundred and twenty-nine children between 6-12 years who were diagnosed with asthma were enrolled. Two hundred and fifty-eight healthy age- and gender-matched children were used as the control group. A questionaire which included 23 factors related to respiratory tract anaphylactic diseases such as residential environment and eating habits were completed by the children's parents.

RESULTSLogistic regression analysis showed that 6 variances out of 16 agents of the residential environment, the experience of raising pets, the type of floor, the type of pillow, the type of quilts, the heating equipments and the house area, were entered into the regression equation; none of the 7 variances of eating inhabits was entered into it.

CONCLUSIONSThe residential environment plays an impotent role in the pathogenesis of bronchial asthma in children. The incidence of bronchial asthma in children can be reduced by the improvement of the residential environment.

Asthma ; etiology ; Case-Control Studies ; Child ; Female ; Humans ; Logistic Models ; Male ; Risk Factors

Objective To evaluate the safety of the split influenza vaccine Anflu.Methods A total of 198 subjects were enrolled in this open clinical trial.Of which,97 aged 18-60 years old and 101 aged 61 years old.28 of the subjects had history of chronic disease,while all the subjects were under stable health condition.After being infoma consented,each subject was vaccinated with one dose of split influeza vaccine and observed for 7 days for adverse reactions.Results A total of 17 subjects reported adverse reactions and the adverse reactions occurrence rate was 8.6%.The adverse reactions occurrence rate was 13.4%in the adult group and 4.0%in the elderly group(x2=5.620,P=0.018).Local adverse reactions occurrence rate was 5.5%and pain at the iniection site was the major symptom.System adverse reactions rate was 3.5% and fever was the most frequently reported symptom.None of the subjects with chronic disease history reported adverse reactions.Conclusion The results showed that the vaccine did not cause new adverse reaction,serious adverse event or rare adverse reaction.The split influenza vaccine was safe.

Objective To evaluate the safety of the split influenza vaccine Anflu.Methods A total of 198 subjects were enrolled in this open clinical trial.Of which,97 aged 18-60 years old and 101 aged 61 years old.28 of the subjects had history of chronic disease,while all the subjects were under stable health condition.After being infoma consented,each subject was vaccinated with one dose of split influeza vaccine and observed for 7 days for adverse reactions.Results A total of 17 subjects reported adverse reactions and the adverse reactions occurrence rate was 8.6%.The adverse reactions occurrence rate was 13.4%in the adult group and 4.0%in the elderly group(x2=5.620,P=0.018).Local adverse reactions occurrence rate was 5.5%and pain at the iniection site was the major symptom.System adverse reactions rate was 3.5% and fever was the most frequently reported symptom.None of the subjects with chronic disease history reported adverse reactions.Conclusion The results showed that the vaccine did not cause new adverse reaction,serious adverse event or rare adverse reaction.The split influenza vaccine was safe.

OBJECTIVETo evaluate the efficacy and safety of oxymatrine in the treatment of chronic hepatitis B.

METHODSA multicenter randomized double-blind placebo-controlled trial was conducted. A total of 144 patients with chronic hepatitis B entered the study for 52 weeks; of them 72 received oxymatrine, and 72 received a placebo. Before and after the treatment, clinical symptoms, liver function, serum hepatitis B virus markers, and adverse drug reactions were observed.

RESULTSIn 144 patients, 14 were dropped and excluded due to inconsistencies in the included standard. Therefore, the efficacy and safety of 130 patients were analyzed. After being treated for 52 weeks, 70.77% of the patients in the study group had a normal ALT level, and in 43.08% and 33.33% their HBV DNA and HBeAg became negative. In the placebo group, 39.68% had normal ALT level, and 12.31% and 3.33% had their HBV DNA and HBeAg become negative. The rates of complete response and partial response in the oxymatrine group were 23.08% and 58.46%, and in the placebo group they were 3.08% and 44.62%. They were significantly higher in the oxymatrine group than in the placebo group. In the oxymatrine treated patients, 12 weeks after its withdrawal, 60.00% had a normal ALT level, 41.54% and 23.33% had both HBV DNA and HBeAg negative. In the placebo group, 31.75% had a normal ALT level, 3.08% and 1.67% had both HBV DNA and HBeAg negative. The rates of complete response and partial response in the oxymatrine group were 21.54% and 47.69%, and in the placebo group they were 0 and 41.54%. They were significantly higher in the study group than in the placebo group. The adverse reaction rates of oxymatrine in the study and the placebo group were 7.69% and 6.15%, respectively, but there was no statistical significant difference between them.

CONCLUSIONOxymatrine is an effective and safe agent for the treatment of chronic hepatitis B.

Adolescent ; Adult ; Aged ; Alkaloids ; adverse effects ; therapeutic use ; Antiviral Agents ; adverse effects ; therapeutic use ; Double-Blind Method ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Quinolizines