Objective:To determine the role of lncRNA TUG1 in pancreatic β cells functioning both in vitro and in vivo.Methods:The lncRNA TUG1 expression in mice pancreas,brain,muscle and other different tissues was examined through qRT-PCR.MTT,flow cytometry,GSIS,ELISA and immunochemistry analyses were performed to detect the effect of lncRNA TUG1 on insulin secretion in vitro and in vivo.Results:lncRNA TUG1 was highly expressed in pancreatic tissue compared with other organ tissues.Knockdown of lncRNA TUG1 expression resulted in decreased insulin secretion in β cells both in vitro and in vivo.Immunochemistry analyses showed decreased relative islet area after treatment with lncRNA TUG1 siRNA.Conclusions:Downregulation of lncRNA TUG1 expression can affect insulin secretion in pancreatic β cells in vitro and in vivo,and lncRNA TUG1 may represent a factor that regulates the function of pancreatic β cells.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Neoplasm Invasiveness ; Skin Neoplasms ; metabolism ; pathology

ObjectiveTo investigate the number of γδT cells in the peripheral blood from patients with systemic lupus erythematosus(SLE) and their correlation to disease activity.MethodsγδT cells were detected in the peripheral blood from 42 SLE patients and 20 normal controls by flow cytometry.Anniex-V/Pl double-staining flow cytometry was employed to observe the proportion of the apoptotic γδ and CD3+ T cells in 6 SLE patients with active disease and 6 normal controls,respectively.The levels of plasma anti-nuclear antibody and anti-dsDNA antibody were determined by using enzyme-linked immunosorbent assay.Data were analyzed with t test and Pearson's correlation test.ResultsThe percentages of γδT cells were remarkably down-regulated in SLE patients [(3.0±1.8)％ ] with active disease compared with that of those patients with inactive[(5.3±3.0)％] disease and normal controls [(6.8±2.8)％](t=-3.071 and -5.913 respectively,both P＜0.01 ).The absolute number of γδT cells decreased significantly in SLE patients with active disease[ ( 1.7± 1.6)× 107/L ] than those with inactive SLE [ (5.3±3.6)× 107/L ] (t=-3.292,P＜0.01 ),and both were lower than the normal controls [ (10.1±5.0)×l 07/L] (t=-7.247 and -2.905 respectively,both P＜0.01 ).There was a negative correlation between systemic lupus erythematosus disease activity index (SLEDAI) andT cell counts in 30 SLE patients with active disease (r=-0.365,P=0.047).γδT cell percentage (r=-0.336,P=0.030) and counts (r=-0.410,P=0.007) were both inversely correlated with the erythrocyte sedimentation rate,but positive correlation were found between hemoglobulln and γδT cell counts (r=0.409,P=0.007).The apoptosis ofγδT cells in SLE patients was more common than in normal controls(t=2.886,P＜0.05 ).The number of apoptotic γδT cells was higher than that of CD3+ T cells in SLE patients (t=2.952,P＜0.05 ).Conclusion γδT cells of the peripheral blood of SLE patients are down-regulated partially due to excessive apoptosis,which may correlate with the disease activity.

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

Normal function of growth hormone-insulin-like growth factor Ⅰaxis is essential for linear growth after birth. A case of continuous growth with undetectable growth hormone level even under insulinhypoglycemia stimulation was reported. The growth hormone deficiency was due to pituitary stalk interruption combined with deficiency of multiple pituitary hormones. Taken together with reviewed literature, this so-called nongrowth hormone-dependent linear growth was preconditioned by other hormones, especially gonadotropin deficiency,and the unclosed epiphysis.

Objective To investigate the effect of interleukin(IL)-21 and dexamethasone(Dex)treatment on the percentage of follicular helper T cells(Tfh)in patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells(PBMCs)obtained from 11 SLE patients were cultured in 96 well plates at 106/well with medium as the control,and with rIL-21(200 ng/μl)or rIL-21(200 ng/μl)+Dex(1×10-6 mol/L)for 24 and 72 hours as the study groups.Another 20 samples were collected and incubated with different concentrations of Dex(0,5×10-7 mol/L,1×10-6 mol/L)for 24 hours.After labeling with CD4,PD1 and CXCR5 monoclonal antibodies,the proportions of Tfh cells were assessed by flow cytometry.ELISA was used to detect supernatant antinuclear antibody(ANA)levels.Differences between groups were analyzed by paired t test.Results The percentages of Tfb cells were elevated when incubated with rIL-21 for 24 h and 72 h[24 h(4.3±1.2)％,(5.9±2.4)％;72 h(5.6±4.0)％,(8.0±5.6)％;t=3.48 and 3.05 respectively,P=0.007 and 0.012].Compared with those in the rIL-21 group,the percentages of Tfh cells in the rIL-21+Dex group were decreased(t=4.70 and 2.78,P=0.001 and 0.019).Tfh cells were decreased ina dose-dependent manner when treated with Dex[24 h(4.2±1.3)％;72 h(6.2±4.4)％;t=3.37 and 2.26,P=0.003 and 0.036].There was no difference of supe(rn)atant ANA levels among those groups.Conclusion IL-21 can promote the proliferation of Tfh cells,while Dex inhibits Tfh cell growth and the inhibition effect is dose dependent.Tfh cells may represent a new target for the treatment of SLE.

Objective To explore the role of GRK5 in sustained β adrenergic receptor (βAR)-stimulated increased levels of oxidative stress.Methods Male SD rats (180-200 g) were separated into 4 groups according to the random principal: control group (CTRL), control with NAC supplement group (CTRL+NAC), ISO treated group (ISO), and ISO treated with NAC supplement group (ISO+NAC), with 6 rats in each group.ISO group was treated by method of intraperitoneal injection for 3 mg/(kg· d).CTRL rats received same volume of physiological saline by same method, while NAC was treated by supplement in drinking water for 15 g/L per day.After 2 weeks of treatment, BP, heart mass index (HMI), histology changes, expression of NOX4 and GRK5 of myocardium was examined.Results HMI of ISO rats was significantly higher than that of the CTRL group [(3.99±0.10 vs 3.31±0.13) mg/g, P＜0.05], and the cardio-myocyte cross-sectional area of ISO group was also significantly increased compared with CTRL group [(11 117.00±387.57 vs 4572.23±176.39) μm, P＜O.05].ISO+NAC significantly reduced the ISO-induced increases of heart weight index (3.56±0.12 mg/g, vs ISO, P＜0.05) and myocyte cross-sectional area (6160.33±141.44 μm2, vs ISO,P＜0.05).The immunohistochemistry results showed that the expression of myocardial NOX4 of ISO group was significantly higher than that of CTRL group [(10.59±1.61 vs 4.35±1.65), P＜0.05], and NAC reduced the ISO induced NOX4 expression increase [(4.67±1.25 vs 10.59±1.61), P＜0.05].Western Blot and immunohistochemistry were used to detect the protein expression of myocardial GRK5.Both results showed that there were no significant differences between ISO and CTRL, ISO+NAC and ISO group (P＞0.05).RT-qPCR detected no significant differences of myocardial GRK5 mRNA expression between ISO and CTRL, ISO+NAC and ISO groups (P＞0.05).Arterial blood pressure showed no significant difference among the 4 groups of rats (P＞0.05).No significant differences were found between rats from CTRL+NAC and CTRL group.Conclusions In the mechanism of sustained βAR-stimulated cardiac hypertrophy, GRK5 may not participate the regulation of hypertrophy-induced factor, and this process needs to be proved in further study.

Objective To investigate the expression and significance of mRNAand exon mutationof NT5C2 gene in acute lymphoblastic leukemia (ALL) bone marrow.Methods Case control study design was used in this study.Bone marrow samples were collected from ALL patients in Anhui Provincial Cancer Hospital in recent 4 years.The patientswere divided into the initial diagnosis group , the complete remission group and the recurrence group.And they could specifically be divided into 36 patients initially diagnosed, 36 patients who achievedcomplete remission and 16 patients who relapsed with children B -ALL,15 patients initially diagnosed,15 patients who achievedcomplete remission and 9 patients who relapsed with children T -ALL, 18 patients initially diagnosed,18 patients who achievedcomplete remission and 12 patients who relapsed with adult B-ALL, and 11 patients initially diagnosed,11 patients who achievedcomplete remission and 6 patients who relapsed with adult B -ALL.The initial diagnosis,complete remission and recurrence samples were matched.8 children and 8 adults without hematologic malignanciewere used as controls .Real-time PCR was performed to detect the level of NT5C2 mRNAin ALL patients.The exons of NT5C2 gene were cloned and sequenced for the common mutations in all cases .The results of NT5C2 mRNA levels in different groups were performed using non -parametric test by SPSS16.0 analytics software, and then non-parametric test together with correlation analysis was analyzed between NT 5C2 mRNA levels of different initial diagnosis groups and gender, age, leukocyte level and risk classification .Results (1)The expression of NT5C2 mRNA levels of recurrence group were higher than that of initial diagnosis group ,complete remission group and controls in children and adult B -ALL respectively(P <0.01).(2)NT5C2 mRNA expression in children and adult T-ALL showed no difference in initial diagnosis ,complete remission, recurrence group and controls (P >0.05).(3)NT5C2 mRNA expression of initial diagnosis group in children and adult B -ALL and T-ALL was not correlated with risk classification (P >0.05).(4)A newheterozygousmutation p.P414A of NT5C2 was discovered in a recurrencesample.Conclusions (1) High expression ofNT5C2 mRNA is associated with recurrence inchildren and adult B-ALL, and it may be an indicator of monitoring recurrence .(2)The incidence of exons mutation of NT5C2 gene in ALL is low in China.

Objective To investigate the effect of early cerebrospinal fluid replacement on nuclear factor-κB (NF-κB) level and clinical outcomes in patients with aneurismal subarachnoid hemorrhage (aSAH) after endovascular embolization.Methods Patients with aSAH received aneurysm embolization were enrolled.They were divided into a cerebrospinal fluid replacement group and a non-cerebrospinal fluid replacement group according to the treatment scheme.All patients were treated with cerebral aneurysm coil embolization within 3 days after admission.The cerebrospinal fluid replacement group performed lumbar puncture cerebrospinal fluid replacement within 24 h after coil embolization,once every other day,20-30 ml of cerebrospinal fluid was replaced each time and 3 mg dexamethasone was injected intrathecally.The NF-κB levels in cerebrospinal fluid were detected at day 1,7 and 14 after the coil embolization.The primary outcome measures were the clinical outcomes determined by the modified Rankin scale (mRS) and the Glasgow outcome scale (GOS) at 3 months after onset.Good outcome was defined as mRS score 0-2 or GOS ＞ 3.The secondary outcome measures included severe complications (hydrocephalus,cerebral vasospasm,cerebral infarction,and rebleeding) and death.Results A total of 81 patients with aSAH received aneurysm embolization were enrolled,including 42 in the cerebrospinal fluid replacement group and 39 in the non-cerebrospinal fluid replacement group.There was no significant differences in the baseline data between the cerebrospinal fluid replacement group and the non-cerebrospinal fluid replacement group (all P ＞0.05).The duration of neck stiffness in the cerebrospinal fluid replacement group was significantly shorter than that in the non-cerebrospinal fluid replacement group (11.3 ± 3.2 d vs.16.5 ± 3.5 d;t =6.985,P ＜ 0.001).The cerebrospinal fluid NF-κB levels were progressively reduced at day 1,7 and 14 after coil embolization in the cerebrospinal fluid rephcement group and non-cerebrospinal fluid rephcement group (all P ＜0.05),but the ccerebrospinal fluid levels of NF-κB in the cerebrospinal fluid replacement group at each time point were significantly lower than those in the non-cerebrospinal fluid replacement group (all P ＜ 0.01).The good outcome rates evaluated according to the mRS score (92.9％ vs.56.4％;x2 =14.446,P ＜ 0.001) and GOS score (97.6％ vs.76.9％;x2 =8.004,P=0.005) in the cerebrospinal fluid replacement group at 3 months were significantly higher than those in the non-cerebrospinal fluid replacement group,and the incidence of cerebral vasospasm was significantly lower than that in the non-cerebrospinal fluid replacement group (14.3％ vs.33.3％;x2 =4.086,P =0.043).Conelusiom Cerebrospinal fluid replacement therapy can reduce the incidence of cerebral vasospasm in patients with aSAH receiving aneurysm embolization and improve clinical outcomes.Its mechanism may be associated with the decrease of NF-κB level in cerebrospinal fluid.