Objective To investigate the role ofglutamic acid decarboxylase autoantibody(GAD-Ab)and protein tyrosine phosphatase autoantibody(IA-2A) in postpartum follow-up of gestational diabetes mellitus (GDM).MethodsGAD-Ab,IA-2A,insulin and glucose metabolism index were measured in 82subjects with normal glucose tolerance (control group) and 84 patients with GDM(GDM group) during 24 to 28 weeks in pregnancy,postpartum 6 to 12 weeks and 2 years.GDM group was divided into antibodies positive group (GAD-Ah or IA-2A were positive) with 18 cases and antibodies negative group (GAD-Ab and IA-2A was negative) with 66 cases.Results Homeostasis model insulin resistance index (HOMA-IR) in GDM group was higher than that in control group (3.87 ± 2.17 vs.2.31 ± 0.52,P ＜ 0.05 ).Homeostasis β -cell function index (HBCI) and 30 min net increment of insulin/30 min net increment of glucose ( △ I30/△ G30) in GDM group were lower than those in control group[206.38 ± 138.06 vs.422.43 ± 228.93 and (20.16 ±11.38) mU/mmol vs.(26.54 ±24.30) mU/mmol,P ＜0.05].The numbers who had the family history of diabetes mellitus and the used of insulin for treatment in antibodies positive group were higher than those in antibodies negative group[ 83.3％ (15/18) vs.28.8％ (19/66) and 77.8％ ( 14/18 ) vs.30.3％ (20/66) ],HOMA-IR,△ I30/ △ G30 and HBCI in antibodies positive group were lower than those in antibodies negative group [3.20±0.84 vs.4.02±0.36,(16.81 ±2.91) mU/mmol vs.(21.55± 11.11) mU/mmol and 124.95 ± 5.03 vs.217.43 ± 115.64,P＜ 0.01 ].Fasting plasma glucose (FPG),2 hours postprandial glucose (2hPG)and glycosylated hemoglobin (HbA1c) in antibodies positive group were higher than those in antibodies negative group during postpartum 6 to 12 weeks and 2 years [postpartum 6 to 12 weeks: (8.20 ±3.11)mmol/L vs.(5.39 ±0.76) mmol/L,(15.22 ±7.29) mmol/L vs.(8.15 ± 1.93) mmol/L,(7.26 ± 1.04)％ vs.(5.88 ±0.41)％ ;postpartum 2 years: (8.91 ±2.80) mmol/L vs.(4.93 ±0.66) mmol/L,(15.75 ±7.87)mmol/L vs.(7.85 ± 1.79) mtmol/L,(7.18 ± 1.22)％ vs.(5.64 ± 0.32 )％,P ＜ 0.01].△ I30/ △ G30 and HBCI were significantly decreased in antibodies positive group postpartum 2 years.No change of the above parameters in antibodies negative group was found.The occurrence rate of type 1 diabetes mellitus (T1DM) was 16.7％(3/18) and 33.3％(6/18) postpartum 6 to 12 weeks and 2 years in antibodies positive group,there was no T1DM in antibodies negative group.ConclusionsWomen with GDM are partly associated with T1DM.Requiring insulin therapy during pregnancy and GAD-Ab or IA-2A positive have considerable risk for developing T1DM.It is also an important predictor to GDM after parturition.

Objective To investigate the correlation of microembolic signals (MES) and outcome in patients with acute ischemic stroke.Methods The patients with acute ischemic stroke were enrolled in the study.The MES of middle cerebral artery was monitored dynamically using transcranial color Doppler ultrasound.The early lesions of ischemic stroke were evaluated by MRI.The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate neurological deficits.The modified Rankin scale was used to evaluate the outcome,and the stroke recurrence was recorded.Results A total of 135 patients with acute ischemic stroke were enrolled,in which,33 were cardiogenic cerebral embolism,49 were large artery atherosclerotic stroke,24 were small arterial occlusive stroke,and 29 were other clear causes or cryptogenic stroke.Multivariate logistic regression analysis showed that coronary heart disease (odds ratio [OR],5.862,95％ confidence interval [CI] 2.008-17.114; P =0.000) was the independent risk factor for positive MES within 48 hours after stroke onset,while the history of antithrombotic treatment (OR 0.376,95％ CI 0.141-0.998; P =0.045) was its independent protective factor.In addition,coronary heart disease (OR 4.879,95％ CI 1.257-18.939; P =0.033),hypertension (OR 4.958,95％ CI 1.029-23.882; P =0.030),and diabetes (OR 3.659,95％ CI 1.027-13.034; P =0.050) were the independent risk factors for positive MES within 1 week after stroke onset.The NIHSS scores of the patients of the positive MES at baseline and 1 week and the clinical outcome at 3 months had no significant differences with the patients of negative MES,however,stroke recurrence and deaths increased significantly (P =0.019).Conclusions MES within 48 hours of onset was not associated with the outcome in patients with acute ischemic stroke at 3 months,however,the incidence of endpoint events such as recurrence and death was significantly higher in patients of positive MES within 3 months.

Objective: To evaluate the clinical efficacy of CAD/CAM glass-ceramic onlays in molars with complicated oblique crown fracture within 3.0 mm subgingival. Methods: Fifty-six molars from 56 patients with complicated oblique crown fracture within 3.0 mm subgingival were recruited after endodontic treatment and divided into 2 groups according to the restorative methods used. The glass-ceramic onlays group was restored with CAD/CAM glass-ceramic onlays, while the all-ceramic crown group received CAD/CAM all-ceramic crowns. The success rates of the restorations were analyzed, and the sulcus bleeding index (SBI), plaque index (PLI), and gingival index (GI) were recorded at the prerestoration and postrestoration stages. Results : After one year of follow-up, the success rate of the glass-ceramic onlay group was 96.4%, and the success rate of the all-ceramic crown group was 92.9%. The difference was not statistically significant (P > 0.05). No differences in SBI, PLI, or GI were found between the glass-ceramic onlay group and the all-ceramic crown group (P > 0.05). Conclusion Conclusion

OBJECTIVETo investigate the effects of storage lesion in apheresis platelets on platelet apoptosis and the changes of aggregation function, and analyze the relationships between the apoptosis and aggregation function.

METHODSPlatelet samples were collected from 10 healthy donors with O blood group. Firstly, the effects of storage lesion in platelets on the platelet apoptosis were detected by flow cytometry. Then, using a multiplate analyzer, individual-donor Plt aggregation response to stimulation by the agonists ADP, Collagen, TRAP and ASPI was examined. Finally, the relationships between its apoptosis and aggregation function was analyzed by correlation regression analysis.

RESULTSBy flow cytometry it was found that with the prolonging of storage time, the apoptosis ratio of platelets significantly increased in a time-dependent manner, the apoptosis rates of platelets on 2nd, 5th and 8th day were (2.87±0.31)%, (11.08±1.54)% and (27.99±2.76)% respectively (P<0.01). Compared with Day 2 platelets, the d 5 platelets stored for 5 d significantly decrease the aggregation response to stimnlation of collagen, TRAP, and ASPI. Compared with platelets stored for 5 d, platelets stored for 8 d significantly decreased the aggregation response to stimnlation of collagen, TRAP and ASPI (P<0.01). However, when stimulated by ADP, the aggregation response was similar among Day 2, Day 5 and Day 8 platelets. The rate of the aggregation function was also declined significantly when stimulated by collagen, TRAP, and ASPI, but not ADP. Further regression analysis showed that the aggregation function of apheresis platelet negatively correlated with the apoptosis (r=-0.9497, r=-0.9527, r= -0.9707 and r= -0.9352 respectively), and the correlation is very strong.

CONCLUSIONWith the prolonging of storage time, the apoptosis ratio in platelets significantly increased. The aggregation function also is declined significantly when stimulated by collagen, TRAP, and ASPI, but not ADP. The aggregation function of apheresis platelets closly correlats with the apoptosis.

OBJECTIVETo study DNA and nucleus damage in human embryo lung fibroblast (HLF) exposed to hydroquinone (HQ) and its genotoxicity.

METHODSHLF were treated with HQ (0, 10, 20, 40, 80 micro mol/L, respectively) for 3 h and DNA damage was detected by comet assay. HLF was also treated with the same concentrations of HQ for 1 h and micronucleus test was performed after they were cultured for 24 h.

RESULTSComet assay showed that percentage of cells with tails in each groups treated with varied doses of HQ was 12%, 19%, 42%, 79% and 95%, respectively, with mean tail length of 7.87, 9.35, 11.03, 19.28 and 23.32 micro m, respectively, in an obvious dose-dependent manner (P < 0.05). Very significant increase in percentage of cells with tails and length of their comet tail were observed in those groups treated with HQ of 20, 40 and 80 micro mol/L (P < 0.01). And, proportion of high and severe DNA damage increased with dose of HQ. HQ could also induce formation of micronucleus and abnormal nucleus in all groups treated by varied doses of HQ, with rates of micronucleus and abnormal nucleus of 2%, 3%, 10%, 9% and 15%, and 6%, 7%, 16%, 27% and 28%, respectively, in a significant dose-dependent manner. There was significant increase in rates of micronuclei and abnormal nuclei in cells treated with HQ at doses of 20, 40 and 80 micro mol/L (P < 0.05).

CONCLUSIONSExposure to HQ could cause DNA and nucleus damage inducing genotoxic effects on HLF.

Cell Nucleus ; drug effects ; Comet Assay ; DNA Damage ; drug effects ; Embryo, Mammalian ; Fibroblasts ; cytology ; Humans ; Hydroquinones ; toxicity ; Lung ; cytology ; Micronucleus Tests

OBJECTIVETo construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA.

METHODSThe conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection.

RESULTS423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%.

CONCLUSIONThe efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully.

DNA Repair Enzymes ; Genetic Vectors ; genetics ; Humans ; Phosphoric Monoester Hydrolases ; genetics ; Plasmids ; RNA, Antisense ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the relationship between the right-to-left shunt(RLS)detected with contrast-enhanced transcranial Doppler (c-TCD) and recurrent stroke in patients with cryptogenic stroke.Methods The consecutive patients w ith ischemic stroke w ere enrol ed. The patients w ith cryptogenic stroke w ere screened according to the TOAST criteria. They w ere divided into either a RLS positive group or a RLS negative group according to the c-TCD findings, and then they w ere fol ow ed up for a period of one year. They w ere also divided into a recurrent group and a non-recurrent group according to w hether they had recurrence or not. Results A total of 118 patients w ith cryptogenic ischemic stroke w ere enrol ed, including 46 in the RLS positive group, 72 in the RLS negative group, 10 in the recurrent group, and 108 in the non-recurrent group. There w ere no significant differences in demographic and baseline data betw een the RLS negative group and the RLS positive group. There w ere significant differences in RLS positive rate (7/10 vs.39/108; P=0.046) and proportion of patients with server RLS (2/10 vs.1/108; P=0.019) betw een the recurrent group and the non-recurrent group. Multivariate logistic regression analysis show ed that the positive RLS w as an independent predictor of recurrent stroke (odds ratio 4.896, 95% confidence interval 1.135-21.120;P=0.033). Conclusions The positive RLS may be an independent risk factor for the recurrence in patients w ith cryptogenic ischemic stroke.

OBJECTIVETo construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair.

METHODSHuman lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed.

RESULTSpEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK.

CONCLUSIONThe hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK.

Antigens, Nuclear ; analysis ; Cell Division ; DNA Damage ; DNA Helicases ; DNA Repair ; DNA-Binding Proteins ; analysis ; deficiency ; Humans ; Ku Autoantigen ; RNA, Antisense ; Transfection

Objective To explore the role of CD4~+CD25~+CD127~(lo) regulatory T cells (Tregs) and inter-leukin (IL)-6, transforming growth factor beta (TGF-β), IL-17 in the pathogenesis of lupus nephritis (LN) by detecting the levels of IL-10, IL-6, TGF-β, IL-17, CD4~+CD25~+CD127~(lo) Tregs in the peripheral blood of patients with active and inactive LN. Methods Three-colour flow cytometry was used to quantitatively measure proportions of Treg cells, the levels of TGF-β, IL-17 were detected by ELISA, and the levels of IL-10, IL-6 in the peripheral blood were detected by Cytometric Bead Array System. Results ① Compared with the inactive LN and the normal controls (P<0.01), the level of CD4~+CD25~+CD127~(lo) Tregs from patients with active LN was lower(P<0.01). When compared with the normal controls, the level of CD4~+CD25~+CD127~(lo) Tregs from LN inactive patients had no significant difference (P>0.05). ② Compared with patients with inactive LN, the levels of IL-10, IL-6 was higher (P<0.01) in patients with active LN. ③ Compared with the patients with inactive LN and the normal controls, the levels of TGF-β, IL-17 was not significantly different (P>0.05). ④ The level of CD4~+CD25~+CD127~(lo) T cell was correlated negatively with the levels of IL-10, IL-6 and SLEDAI (P<0.05), and was not correlated with C3 and C4. ⑤ SLEDAI was correlated positively with the levels of IL-10 and IL-6 (P<0.01). SLEDAI and the level of IL-10 were correlated negatively with C3 and C4 (P<0.01 for both). ⑥ The level of CD4~+CD25~+CD127~(lo) Tregs from LN was not correlated with TGF-β and IL-17. ⑦ TGF-β was correlated positively with the level of IL-17. Conclusion ① The level changes of Tregs and IL-10, IL-6, TGF-β in the peripheral blood of LN can be used as the indicators for the activity status of lupus nephritis. ② Tregs and IL-10, IL-6 in the peripheral blood of LN patients is negatively correlated. ③ The glucocorticoid hormone is helpful to elevate the level of Tregs but decrease IL-17. T cell level can vary in different body status, different microenvironmental and immune status.

Adult ; Carbolines ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Middle Aged ; Phosphodiesterase 5 Inhibitors ; therapeutic use ; Piperazines ; therapeutic use ; Purines ; therapeutic use ; Semen ; Sildenafil Citrate ; Specimen Handling ; Sulfones ; therapeutic use ; Tadalafil