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.

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.

Objective To investigate the effect of downregulation of KIAA0101 protein expression on the proliferation and invasion of a cutaneous squamous cell carcinoma cell line SCL-1,and to explore possible molecular mechanisms underlying the effect.Methods SCL-1 cells were classified into three groups: siRNA control group transfected with the control siRNA,KIAA0101 group transfected with KIAA0101 siRNA,and untreated group remaining untreated.After additional culture,Western blot was used to detect the expression of KIAA0101 protein and proteins associated with cell proliferation and invasion,cell counting kit-8 (CCK-8) to evaluate cellular proliferative activity,and Boyden chamber assay to estimate invasive ability of cells.Results The relative expression level of KIAA0101 protein was 0.062 ± 0.095 in the KIAA0101 group,significantly lower than that in the untreated group (0.359 ± 0.044,P ＜0.05) and siRNA control group (0.379 ± 0.025,P ＜0.05).A significant decrease was observed in cellular proliferative activity (from 24 to 96 hours) and invasive activity (at 48 hours) in the KIAA0101 group compared with the other two groups (all P ＜0.05).Moreover,compared with the untreated group and siRNA control group,the KIAA0101 group showed a stronger expression of p21 protein (0.570 ± 0.060 vs.0.048 ± 0.018 and 0.055 ± 0.014,P ＜0.01) but a weaker expression of matrix metalloproteinase 2 (MMP2) protein (0.051 ± 0.013 vs.0.205 ± 0.029 and 0.221 ± 0.029,P ＜0.01).Conclusion The inhibition of SCL-1 cell proliferation and invasion induced by the downregulation of KIAA0101 gene expression may be associated with the expression changes of p21 and MMP2.

Objective:To evaluate the expression of CXCR4 and the migration rate of bone marrow stromal CD34+cells in differ-ent risk groups with myelodysplastic syndromes (MDS) using correlation analysis. Methods: Forty MDS patients were divided into low-and high-risk groups based on the International Prognosis Scoring System (IPSS). The former was composed of 20 patients with IPSS<1.5, whereas the latter was composed of 20 patients with IPSS≥1.5. Bone marrow (BM) samples of these patients and 10 nor-mal controls were collected. CD34+cells were separated and purified. The expression of CXCR4 was determined by flow cytometry. The migration rate of CD34+cells on the chemotactic effect of SDF-1αand on the effect of bone marrow stromal cells were measured. Results:The expression rate of CXCR4 was higher in the high-risk MDS group than in the low-risk and control groups (P<0.000 1). No significant differences existed between the low-risk and the control groups (P>0.05). The migration rate of CD34+cells on the ef-fects of SDF-1αand marrow stromal cells were significantly increased in the high-risk MDS group compared with those in the low-risk and control groups (P<0.000 1). Migration rate of CD34+cells on the effect of marrow stromal cells was positively correlated with CX-CR4 expression (P=0.000 1). Conclusion:The CXCR4 expression and migration rates of CD34+cells on the effect of marrow stromal cells are significantly higher in the high-risk MDS group than in the low-risk group. Migration rate has a positive correlation with the CXCR4 expression, which further indicates that MDS is a heterogeneous group of hematopoietic stem cell malignancies. The expres-sion and function of SDF-1 and its receptor CXCR4 differ within each group with various risks. SDF-1 and CXCR4 may be involved in MDS pathogenesis.

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 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.

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

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 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 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.