Objective To investigate the effect of bcl-2 gene tr ansfection on the apoptosis and bioactivity of cultured islet cells. Met hods Adenovirus-mediated gene transfection was employed to transfect b cl-2 into the islet cells, the expression of bcl-2 was determined by RT-PCR a nd immunocytochemical staining, and the apoptosis of islet cells was detected by TUNEL, the viability was determined by trypan blue staining and islets function was evaluated by insulin detection. Results Bcl-2 protein exp ressed in 70% of transfected rat islet cells. The apoptosis rate of transfected cells reduced significantly compared with untransfected cells (6% vs. 22%). The viability of transfected cells was higher than that of untransfected cells (91% vs. 68%). Insulin secrete capacity of transfected islet cells in high glucose me dium was superior to untransfected cells. Conclusions Islet cel ls may be successfully transfected with adenovirus-mediated gene transfection t echniques. The apoptosis may be declined by bcl-2 gene transfection, so the isl et cells' function improved.

Diagnostic Errors ; Female ; Humans ; Infant, Newborn ; Inhalation ; Male ; Organophosphate Poisoning ; Poisoning ; diagnosis

Acute Disease ; Adult ; Ethylene Oxide ; poisoning ; Female ; Humans ; Male ; Middle Aged

AIM: To explore correlation,consistency and comfort between traditional tear film examination methods and Oculus Keratograph.METHODS: A retrospective study.Totally 101 cases (101 eyes) were diagnosed myopia and then accepted LASEK (laser epithelial keratomileusis).Non-invasive tear film break-up time(NIBUT),lower tear meniscus height(LTMH) were measured with Oculus Keratograph,fluorescein tear film break-up time(fl-BUT) and Schimer Ⅰ test (SⅠt) were performed on all cases.The correlations analysis between NIBUT and fl-BUT,LTMH and SⅠt were performed by Spearman rank correlation,consistency check between NIBUT and fl-BUT by Bland-Altman analysis.Visual analogue scale(VAS) was applied on evaluating the comfort of two kinds of examination methods.RESULTS: LTMH and SⅠt showed positive correlation (rs=0.346,P=0.001).NIBUT and fl-BUT showed positive correlation (rs=0.393,P=0.001),95% consistency limits range-9.62 to 14.18 in Bland-Altman Figure.There was significant difference between VAS of NIBUT and VAS of fl-BUT(z=-2.324,P=0.020).There was significant difference between VAS of LTMH and VAS of SⅠt (z=-8.845,P=0.001).CONCLUSION: Oculus Keratograph can objectively measure NIBUT and LTMH,and was more comfortable than traditional tear film examination methods.It can effectively assess tear film function before corneal refractive surgery.

ObjectiveTo evaluate the effects on serum homocysteine(Hcy) level,vascular function and carotid intima-media thickness(IMT) in metformin-treated diabetic patients with or without supplementation with folate and Vitamin B12.MethodsA total of 100 newly diagnosed diabetic type 2patients were divided into two groups by random digits table with 50 cases each,90 patients completed study.Forty-seven participants (control group) received a 6-month course of metformin treatment,43 patients (treatment group) received mefformin,folate and Vitamin B12 treatment.The levels of serum Hcy,endothelin-1 (ET-1),carotid IMT,large or small arterial elasticity index (C1,C2),flow-mediated dilatation (FMD) of the brachial artery were evaluated before and after treatment.ResultsThe level of serum Hcy in control group significantly increased compared with before treatment[ (13.4 ± 2.7)μ mol/L vs.(11.1 ± 1.9)μ mol/L],hut the level of serum Hcy in treatment group significantly decreased compared with before treatment [ (9.2 ± 1.8 ) μ mol/L vs. ( 11.3 ± 2.0) μ mol/L ],there was significant difference(P ＜ 0.05 ).A beneficial effect was observed in the serum ET-1,FMD,carotid IMT and C2 in treatment group[ (20.0 ± 6.2)ng/L,( 15.8 ± 7.6)％,(0.8 ± 0.2) mm,(4.1 ± 2.1 ) ml/mm Hg ( 1 mm Hg =0.133 kPa) × 100 vs. (31.3 ±10.1 ) ng/L,(9.7 ± 4.5)％,( 1.1 ± 0.4) mm,(2.3 ± 1.0) ml/mm Hg × 100 ] (P ＜ 0.05).The levels of ET-1,FMD,carotid IMT and C2 after treatment in control group [ (24.8 ± 6.8) ng/L,( 12.9 ± 6.3 )％,(0.9 ± 0.3)mm,(3.0 ± 1.4) ml/mm Hg × 100] had significant difference compared with before treatment [ (30.6 ± 8.7)ng/L,(9.8 ± 4.6)％,( 1.0 ± 0.3) mm,(2.2 ± 0.9) ml/mm Hg × 100](P＜ 0.05).However,the results were improved significantly in treatment group than those in control group (P ＜0.05).In treatment group,significant correlation were detected between changes of Hcy and ET- 1 (r =0.43,P ＜ 0.05 ),carotid IMT(r =0.56,P ＜ 0.05),FMD (r =-0.54,P ＜ 0.05 ),C2 (r =-0.37,P ＜ 0.05 ).ConclusionsAdministration of folate and Vitamin B12 can reduce the levels of serum Hcy and ET-1 in metformin-treated type 2 diabetic patients,which exert beneficial effect on carotid IMT,FMD and C2.

Objective To investigate the impact of experimental stress on serum glucose , insulin, glucagon, cortisol, and ep-inephrine in diabetic rats and its intervention with insulin and fluoxetine .Methods Type 2 diabetic rats model was induced by feeding with high sugar and high fat diet , and intraperitoneal injection of streptozotocin (STZ).The rats were divided into normal group , dia-betic control group , diabetic treated with insulin , and diabetic treated with fluoxetine .All the rats were exposed to multiple stressors (forced swimming, cold stimulation, rotation, restrain, and crowding) for 8 weeks.The rat blood sera collected from each group were analyzed with multiple parameters including glucose , insulin, glucagon, cortisol, and epinephrine at the 4th and 8th week.Results After experimental stress, the levels of glucose [(16.7 ±3.5)mmol/L vs (5.1 ±1.1)mmol/L, t =13.9, P <0.01], glucagon [(158.5 ±50.2)ng/L vs (120.8 ±38.7)ng/L, t =2.5, P <0.05], epinephrine [(203.8 ±48.6)pg/ml vs (158.7 ±42.6)pg/ml, t =2.9, P <0.01], cortisol [(21.3 ±4.8)ng/ml vs (18.2 ±3.8)ng/ml, t =2.1, P <0.05], and HOMA-IR (10.9 ±2.6 vs 3.3 ±0.8 , t =12.3 , P <0.01 ) of diabetic rats were significantly increased .The glucose level [ ( 9.7 ±2.1 ) mmol/L vs ( 16.7 ± 3.5)mmol/L, t =7.0, P <0.01] was significantly improved after treated with insulin .The levels of epinephrine [(158.8 ±37.5 ) pg/ml vs (203.8 ±48.6)pg/ml, t =3.0, P <0.01], and cortisol [(15.7 ±4.2)ng/ml vs (21.3 ±4.8)ng/ml, t =3.5, P <0.01 ] were significantly improved after treated with fluoxetine .Conclusions Experimental stress increased the levels of glucose , en-docrine hormone , and insulin resistance of diabetic rats .Treatment with insulin improved the glucose level and treatment of fluoxetine improved the endocrine hormone level .

Murine cytomegalovirus (MCMV) IE3 protein is a multifunctional viral protein that interacts with several target proteins of both viral and host cellular origin. To investigate the biological function of IE3 in the pathogenesis of the brain disorders caused by CMV, a screening for host cellular proteins that could interact with IE3 was performed. By yeast two-hybrid screening, ankyrin repeats domain 17 (Ankrd17, also known as Gtar) was identified as a host factor that could interact with IE3. This interaction was verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion. Mapping analysis suggested that the 1-148 residues of IE3 were responsible for the interaction. These results suggested that the interaction between Ankrd17 and IE3 may play a key role in the pathogenesis of MCMV-associated disease.

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

BACKGROUND:The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.
OBJECTIVE:To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.
METHODS:THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups:mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. cells in each group were col ected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups:HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cellnumber was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.
RESULTS AND CONCLUSION:After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly;compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100%2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

Objective To investigate the effects of antithyroid drug(ATD) on oxidative stress in Graves disease patients with and without infiltrative ophthalmopathy.Method The levels of superoxide dismutase(SOD),glutathione peroxidase(GPx) and catalase(CAT) in erythrocytes of Graves disease patients(18 with and 20 without infiltrative ophthalmopathy) were measured before and after treatment with ATD.Results Compared with the normal control group,the levels of SOD,GPx and CAT were significantly higher in Graves disease patients with and without infiltrative ophthalmopathy.After euthyroidism was achieved with the treatment of ATD,the levels of SOD,GPx and CAT were normalized in the patients without infiltrative ophthalmopathy,but oxidative stress was still present in the patients with infiltrative ophthalmopathy.Conclusion The oxidative stress is obvious in patients with Graves ophthalmopathy,which suggests that oxidative stress is involved in orbital inflammation.