Objective To investigate the effect of angiotenisn ⅡI (Ang Ⅱ) on RIN-m β-cell,and to explore the mechanism of β-cell function impairment caused by Ang Ⅱ.Methods RIN-m cells were cultured with various concentrations of AngⅡ (0.1,1,10,100 nmol/L).After incubation for 24 hours,the basal(3.3 mmol/L) and glucose-stimulated(16.7 mmol/L) insulin secretion(GSIS)were detected by radioimmunoassay,mRNA and protein expressions of uncoupling protein 2(UCP2)were determined by RT-PCR and Western blot,respectively.The intracellular ATP content was measured by luciferase bioluminescence.The mitochondrial membrane potential and cellular Ca~(2+) concentration were detected by flow cytometry.Results (1) Various concentrations of Ang Ⅱ had no significant influence on the basal insulin secrection of RIN-m cell(F=0.644,P = 0.634).Except for 0.1 nmol/L AngⅡ,the other concentrations of Ang Ⅱ markedly reduced GSIS of RIN-m cells(F= 118.528,P = 0.000).(2) Compared with the control group,Ang Ⅱ significantly increased mRNA and protein expression of UCP2(F= 1 370,P = 0.000;F=675.175,P = 0.000).(3)Except for 0.1 nmol/L Ang Ⅱ,the other concentrations of Ang Ⅱ significantly decreased the mitochondrial membrane potential,cellular ATP content,and cellular Ca2+ concentration of RIN-m cell(F=4.035,P=0.008;F=3.353,P = 0.013;F=5.867,P = 0.001).Conclusion Ang Ⅱ impairs GSIS of p-cell,the mechanism of impairment may be interpreted that Ang Ⅱ can increase the expression of UCP2,furthermore,it can reduce mitochondrial membrane potential,decrease the content of cellular ATP and the concentration of cellular Ca~(2+),can finally impair the function of β-cell.

Humans ; Hypoxia-Ischemia, Brain ; pathology ; Infant, Newborn ; Magnetic Resonance Imaging

Objective To investigate the effect of angiotensin Ⅱ(AngⅡ) and losartan on ? cells,and its related molecular mechanisms.Methods Normally cultured RIN-m cells were divided into three groups:control group(cultured with RPMI 1640 medium),AngⅡ group(treated with 100 nmol/L AngⅡ),losartan group(treated with 1 ?mol/L losartan 15min before addition of 100nmol/L AngⅡ).After incubation for another 48h,the apoptosis rate of RIN-m cells was quantified by flow cytometry(FCM) with Annexin-V FITC/PI dual staining.The expressions of Bcl-2 and Bax mRNA and protein were determined by RT-PCR and Western blotting,and the activities of Caspase 3 and Caspase 9 were detected by spectrophotometry.Results The apoptosis rate of RIN-m cells was significantly higher in AngⅡgroup than in both control and losartan group(P0.05).In comparison to the control group,the expressions of Bcl-2 mRNA and protein significantly declined,while of Bax mRNA and protein increased obviously in AngⅡ group(P0.05),but the expressions of Bcl-2 mRNA and protein were significantly higher(P0.05).Conclusion AngⅡ may induce the apoptosis of ? cells through mitochondrial pathway,and pre-intervention with losartan,which partly reverses the effect of AngⅡ,may play a protective effect on ? cells.

Objective To explore the value of dexamethasone(DEX) for neuronal cell injury and death by observing the effect of DEX on excitatory amino acid(EAA) and monoamine neurotransmitter in cerebral tissue of neonatal rat with hypoxia-ischemia.Methods Hypoxic-ischemic neonatal rat models were established,the levels of EAA and monoamine neurotransmitter in cerebral tissue were analyzed by using capillary electrophoresis and fluorospectrophotometry method.The rats were divided into 4 groups: small dose DEX group pre-treated with DEX(0.5 mg/kg) prior to hypoxia-ischemia,large dose DEX group pre-treated with DEX(10 mg/kg) prior to hypoxia-ischemia,HIE group and shamful operation group.Results The levels of EAA and monoamine neurotransmitter contents in HIE group were significantly higher than those in shamful operation group(P0.05).EAA contents of large dose DEX group greatly decreased compared with HIE group (P

Objective To explore the influence of type 2 diabetes mellitus combined with subclinical hypothyroidism on diabetic vascular complications.Methods One hundred and two patients with type 2 diabetes mellitus were selected.The serum free triiodothyronine (FT3),free thyroxine (FT4),thyroid stimulating hormone (TSH),anti-thyroid peroxidase antibody (TPO-Ab),thyroglobulin antibody (TG-Ab) levels were measured by chemiluminescence method.The patients were divided into type 2 diabctes mellitus combined with subclinical hypothyroidism group (47 cases) and type 2 diabetes mellitus with normal thyroid function group (55 cases) according to the thyroid function.The glycated hemoglobin (HbA1c),total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C),triacylglycerol (TG),high-density lipoprotein cholesterol (HDL-C),urea nitrogen,creatinine and albumin levels were measured.The estimated glomerular filtration rate (eGFR) was calculated according to the formula of modification of diet in renal disease (MDRD).The presence of diabetic retinopathy was examined by fundus examination,and the presence of lower limb artery lesions was measured by vascular ultrasound.All indicators were compared between 2 groups.Results There were no statistical differences in age,disease course,HbA1c,body mass index (BMI),TC,TG,HDL-C,LDL-C,incidence of lower limb artery lesions and incidence of diabetic retinopathy between 2 groups (P＞ 0.05).TheeGRF in type 2 diabetes mellitus combined with subclinical hypothyroidism group was significantly lower than that in type 2 diabetes mellitus with normal thyroid function group:(83.74 ± 21.55) ml/(min· 1.73 m2) vs.(115.02 ± 12.29) ml/(min· 1.73 m2),and there was statistical difference (t =4.274,P ＜ 0.01).The incidence of diabetic nephropathy in type 2 diabetes mellitus combined with subclinical hypothyroidism group was significanlty higher than that in type 2 diabetes mellitus with normal thyroid function group:48.9％ (23/47) vs.23.6％(13/55),and there was statistical difference (x2 =7.103,P＜ 0.01).Logistic regression analysis showed that subclinical hypothyroidism was a risk factor for diabetic nephropathy (OR =0.524,95％ CI 0.12-0.93,P ＜ 0.05),but it was not the risk factor for diabetic retinopathy (OR =0.618,95％ CI0.19-2.16,P =0.475) and lower limb artery lesions (OR =0.485,95％ CI 0.32-2.13,P =0.689).Conclusion Subclinical hypothyroidism in patients with type 2 diabetes mellitus has no obvious effect on lower limb arterial complications and diabetic retinopathy,but may increase the risk of diabetic nephropathy.

Objective To investigate the curative effect of modified tracheal catheter in acute respiratory failure caused by central airway stenosis.Methods 16 cases inpatient with acute respiratory failure caused by central airway stenosis were involved.Found out the position and range of stenosis of central airway by X-ray and CT of chest and fiberbronchoscope,chose the suitable silicon suction tube and cut it to make a tracheal catheter,then guided the catheter through the stenosis by fiberbronchoscope to construct artificial airway.Results The dyspnea of all 16 cases of acute respiratory failure caused by central airway stenosis could by relieved in short time,the PaO_2 raised from(39?12)mm Hg to(72?10)mm Hg,SaO_2 raised from(75?13)% to(93?3)%,PaCO_2 dropped from(102?21)mm Hg to(62?13)mm Hg after therapy.The effective rate is 100%.There was no other serious complication except for 2 cases of little amount of bleeding in trachea.15 cases survived and one died of serious muhisystem organ failure.Conclusions The use of modified tracheal catheter in treatment of acute respiratory failure caused by central airway stenosis can relieve the acute dyspnea in short time,it also can dilate central airway,save the cost of tracheal balloon dilatation for the follow-up therapy.

Objective To explore molecular mechanism and the curative effect of Yisuishengxue powder and its function of the hepersensitive site 2 (HS2) in ?-globin gene cluster locus control region binding with nucleoprotein.Methods After 3 months treatment of Yisuishengxue powder, nucleoprotein was extracted from the morrow cell before and after treatment. The HS2 DNA probes was combined with nucleoproteins.Electrophoresis gel mobile lag was utilized for observing the mobile velocity of DNA segment.Observe the mobile velocity of DNA probes.Results The mobile velocity of probes combined with nucleoproteins before treatment was different form that of the controls, while it was very close to the controls after treatment.Conclusions It is suggested that this compounding medicine might affect the DNA segment of HS2 site in ?-LCR binding with nucleoprotein GATA-1, which may be one molecular mechanism of Chinese herb therapy.

Objective To investigate histopathological changes and expression of integrin ?1 of sternomastoid muscle,and probe the mechanism and significance during disease process in congenital muscular torticollis(CMT).Methods Histopathological changes of sternomastoid muscle section stained with HE and Gomori silver staining were observed and the expression of integrein ?1 with immunohistochemistry was detected,and the expressive quantity and distribution with image analysis system was quantitive analyzed.Results 1.With light microscopy observation,the results showed that the fibrous degeneration of sternomastoid muscle could be summed up 2 kinds: A category displayed the myocytes atrophyed,and there were lots of connective tissue hyperplasy around myocytes,and the direction of fibrous arrangement was disordered,meanwhile there were lots of vessels and nervers hyperplasy,and eventually the myocytes shrank back and disappeared.B category displayed that the structure of cross striation or sarcomere disappeared or changed,and myocytes could maintained the outline and the sarcolemma were integrated,and then fibrous pathological changes of myocyte took place,and there were lots of fibroblast-like that had much more enations between fiber bundles.With Gomori silver staining,the major changes of fibrotic sternomastoid muscle showed that there were lots of collagenous fibers hyperplasy.The arrangements of collagenous fibers were disordered in A category and were well-arranged in B ca-tegory.2.With immunohistochemistry,the results showed the expression of integrin ?1 was weak positive in normal control group(125.7?5.167).In diseased groups,the results showed 3 different extents:the expression of integrin ?1 displayed stronger positive in A category myocytes(30.15?6.543),and the level of expression was significantly different from normal controls(P0.05).Conclusions The fibrous pathological changes of sternomastoid muscle are a complicated and gradually process,which may has different mode,and ingetrin ?1 may participated the process of pathological changes.

Objective To explore a method for isolation and culture of human epidermal stem cells. Methods Epidermis was obtained by digesting human foreskin with Dispase Ⅱ and Trypsin-EDTA.After suspension on the epidermal stem cell medium (ESCM), these single epidermis cells were inoculated onto human collagen Ⅳ-coated flasks and cultured at 37 ℃ in a humidified atmosphere containing 5% CO_2 for 10 min. The nonadherent cells were rinsed off 10 min after inoculation, and the adherent cells continued to be cultured after enriching and abstraction by type Ⅳ collagen. The cell growth was observed through inverted microscope, and the cell cloning efficiency and time of clone sustain were also detected. Immunocytochemistry was used to observe the expression of ?_1-integrin and keratin 19(K19). Keratinocytes were served as controls. Results It was revealed by histological observation that colonies were formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher and the time of clone sustain was longer than that of the control group. Positive expression of ?_1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion Adult epidermal stem cells could be successfully isolated and cultured by adhension with type Ⅳ collagen and culture with ESCM.

Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua.