Objective:To clarify the mechanism of immunoregulation of bone marrow stromal cells(BMSCs) transfected interleukin 18(IL-18) after their transplantation into glioma bearing rats.Methods:Cultured BMSCs from SD rats were transfected with rmIL-18(BMSCs/IL-18).Untransfected BMSCs were used as control.Culture supernatant medium was collected for IL-18 examination at different time point by ELISA kit.After establishment of glioma bearing rats followed by BMSCs/IL-18 transplantation,serum concentration of IFN-?,IL-2 and IL-10 were examined by means of ELISA kit.And their splenocytes were cultured with C6 cells and BMSCs/IL-18 for in vitro cytotoxicity assay,and subsets of splenocyte were detected by flow cytometry.TUNEL was used to clarify apoptosis cells inside glioma and anti-CD34 staining was performed to observe microvessel density(MVD).Results:BMSCs/IL-18 could secret IL-18 long term and stably.After being transplanted with BMSCs/IL-18,serum concentration of IL-2,IFN-? in glioma bearing rats' increased obviously and serum concentration of IL-10 decreased.Flow cytometry results showed that CD4+ and CD8+ T lymphocytes increased in the splenocytes.And rechallenge with C6 cells induced a rapid immuno-reaction.In vitro cytotoxicity assays.It was showed that BMSCs/IL-18 could stimulate splenocytes to kill C6 cells obviously.TUNEL assay showed that there were 15.74?6.23 apoptosis cells inside glioma in each view in Group 2,which was much more when compared with other groups.Microvessel density inside glioma in group 2(6.51?2.71) was lower than in group 1(13.52?3.06),group 3(12.67?2.61) and control group(14.84?1.47).Conclusion:By means of inducing Th1 cytokine and suppressing Th2 cytokine and activating cytotoxic T lymphocyte,BMSCs/IL-18 induces obviously anti-tumor activity.

Objective To investigate the clinical significance and abnormal expression of the CREB in different grade gliomas. Methods The expression of CREB was examined by using immunohistochemistry in brain tissues from the brain injury (5 cases) and different grade gliomas (55 cases).The mRNA and protein levels of CREB were further as?sessed using Western blot and RT-PCR in brain tissues from the patients with brain injury (10 cases) and those with dif?ferent grade gliomas (30 cases). Results The positive rates of CREB immunohistochemistry were 2/5 in control, 10/15 inⅠ-,Ⅱ11/12 in Ⅲ, 28/28 in Ⅳ. The positive rates of CREB were significantly different among different groups (H=28.183,P<0.05).The mRNA levels of CREB were 1.00 ± 0.000 in control, 1.35 ± 0.068 inⅠ-Ⅱ, 2.88 ± 0.111 in Ⅲand 3.75 ± 0.196 in Ⅳ. The expression of CREB was higher in the glioma than in control group, and the mRNA levels of CREB were significantly different among different groups(F=1.208,P<0.05). The protein levels of CREB were 0.311 ± 0.014 in control, 0.469±0.026 inⅠ-Ⅱ, 0.641±0.028 inⅢand 0.896±0.024 inⅣ. The protein levels of CREB were sig?nificantly different among different groups(F=1.123,P<0.05). Conclusion The expression of CREB is elevated in glio?mas with different differentiation degrees. The expression of CREB was positively correlated with the degree of differentia?tion, indicating that CREB may have an important regulatory role in the progress of gliomas.

Objective To investigate the diagnostic value of carotid artery ultrasonography,CT angiography (CTA)and digital subtraction angiography (DSA)for carotid artery dissection. Methods The image data of carotid artery ultrasonography,CTA,and DSA of 24 patients with carotid artery dissection were analyzed retrospectively. Results Twenty-four,16,and 21 patients were examined with DSA,CTA,and carotid artery ultrasonography respectively. The detection rates of carotid artery dissection with DSA,CTA, and carotid artery ultrasonography were 95. 8%,75.0%,and 71. 4% respectively. The DSA mostly showed the line-like sign (n=12,50 %). CTA and carotid artery ultrasonography mostly showed the double lumen sign;they were 37. 5%(n=6)and 52. 4%(n=11)respectively. Compared with DSA,the concordance rates of carotid artery ultrasonography and CTA were 66. 7% and 81. 3% respectively. There was no significant difference (Kappa=0. 39,P=0. 08 and Kappa=0. 43,P =0. 22 respectively). The concordance rate of ultrasonography in combination with CTA and DSA reached 87. 5%(n=15,Kappa=0. 67,P =0.047). There was significant difference. Conclusion DSA is a gold standard for the diagnosis of carotid artery dissection,and it is irreplaceable. Carotid artery ultrasonography in combination with CTA can improve the diagnostic rate. Carotid artery ultrasonography can be used as a screening method for carotid artery dissection.

Objective Long noncoding RNA (lncRNA) has been identified to regulate DNA methylation, histone acetylation, and gene post-transcriptional regulation in kinds of diseases, including tumorigenesis, obesity and so on.Therefore, lncRNAs might be the potential targets of mesangial cells proliferation.Methods Mesangial cells were exposed to suitable concentration of TGF-β through cell proliferation assay;then the lncRNAs expression levels were detected by microarray in experimental group and control group separately;finally the differentially expressed lncRNAs were identified by RT-PCR;meanwhile, and the expression levels of target genes were also detected by RT-PCR.Results Cell viability assay confirmed that 10 ng/ml TGF-β could promote mesangial cell proliferation significantly.Totally, over 30 000 lncRNAs were detected in TGF-β treated MCs and control group cells separately.Compared to the control group, 5550 lncRNAs differentially expressed in TGF-β treated MCs, including 119 up-regulated and 147 down-regulated over 2 fold.RT-PCR results appeared that uc.60, MRAK079149, MRAK029456, XR_005507, XR_007641, uc.14, and uc.412 were significantly up-regulated in TGF-β treated MCs, and BC088254, DQ402472, BC098733, BC158832,BC098746 were stably down-regulated.Compared to the control group, the mRNA expression levels of AATF and NEK were increased in the TGF-β treated mesangial cells (P ＜ 0.05).AATF and NEK were downstream target genes of uc.412 and MRAK079149 respectively.Conclusion The differential expression of long noncoding RNAs presents in the experimental mesangial cells proliferation induced by TGF-β.

Objective To explore the methods and the nursing points during the course of using methotrexate to cure moderate and severe psoriasis.Methods 41 moderate and severe patients with psoriasis have been adopted in this study.The procedure of nursing care included medical education,clinical care and psychological nursing in order to increase the compliance of patients.Results All the patients have accepted the therapy of using methotrexate to cure psoriasis,the effective rate was 80.5%,only one case have withdrawed from the study because of leucopenia.The compliance of patients have increased significantly.Conclusion Using methotrexate to cure psoriasis is effective and safe,which can increase the compliance and the confidence of patients.

Objective To study the effect of paeonol (PAE) and panax notoginseny saponins (PNS) on the expressions of collagenⅠandⅢprotein and mRNA in rats with acute myocardial infarction (AMI), and to explore the molecular mecha?nism of improving myocardial fibrosis. Methods The rat model of AMI was made using the left anterior descending coro?nary branch ligation.According to the intervention rats were divided into model group, PAE (8 mg/kg) group, PNS (40 mg/kg) group, PAE (4 mg/kg)+PNS (20 mg/kg) group, PAE (8 mg/kg)+PNS (40 mg/kg) group and captopril positive control group (10 mg/kg). Sham operation group, only wear line without ligation. The left ventricular mass index (LVMI) was detected after treatment for 28 d. Masson staining was used to observe changes of myocardial fibrosis. Western blot assay and RT-PCR technique were used to detect protein and mRNA expression levels of collagenⅠandⅢ. Results The values of LVMI were increased in model group compared with those of sham operation group and treatment groups. Compared with PAE group and PNS group, values of LVMI were significantly decreased in PAE (4 mg/kg)+PNS (20 mg/kg) group and PAE (8 mg/kg)+PNS (40 mg/kg) group. There was a more significant decrease in formula high dose group (P < 0.01). The model group showed pathological change. There were different degrees of improvement in pathological structure in all treatment groups, more sig?nificant improvement was found in formula low dose group, formula high dose group and captopril positive control group. There were different degrees of increase in expressions of collagenⅠandⅢprotein and mRNA in model group compared with those of sham operation group and treatment groups. Compared with PAE group and PNS group, the protein and mRNA expression levels of collagenⅠandⅢwere significantly decreased in formula low dose group and formula high dose group,more significant decreased was found in formula high dose group (P<0.05). Conclusion Compound of paeonol and PNS can improve myocardial fibrosis in myocardial infarction rats, which may be related with reduced expressions of collagenⅠandⅢprotein and mRNA.

Objective To investigate the effect of immune modulation therapy on heart function and cytokines in elder patients with chronic heart failure (CHF). Methods The 96 patients aged 60-78 years with New York Heart Association(NYHA)functional. class Ⅱ-Ⅳ CHF were randomly divided into two groups: CHF treatment group received regular therapy and thymopetidum and CHF control group received regular therapy. Another 45 healthy individuals aged 60-80 years were involved as normal control. The ejection faction of left ventricle (LVEF), inflammatory cytokines including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), anti-inflammatory cytokine interleukin-10 (IL-10), plasma high sensitive C-reactive protein (hsCRP), plasma brain natrium peptide (BNP)and Minnesota Living with Heart Failure Questionnaire (LHFQ) assessment were tested before therapy, 15 days and 75 days after treatment. Results (1) Before therapy, compared with normal control group, the levels of TNF-α, IL-1β, TNF-α/IL-10 ratio, BNP, hsCRP and LHFQ were significantly increased (P ＜ 0. 05 or P ＜ 0. 01 ), and the levels of IL-10, LVEF were markedly decreased (P＜0.01) in the patients of CHF treatment group and CHF control group. While no difference between the two CHF groups was observed. (2) After the first course of treatment,compared with CHF control group, the levels of IL-10 were increased (P＜0. 01), while the levels of TNF-α, IL-1β, BNP and hsCRP were decreased (P＜0.05 or P＜0.01) in CHF treatment group. The level of LVEF was increased, TNF-α/IL-10 ratio (4.84 ±0. 53 vs. 5.28±0. 66) and LHFQ were decreased even though there was no significant difference between the two groups. (3) After the second course of treatment, compared with CHF control group, the levels of IL-10 and LVEF were increased (P＜0. 05 or P＜0.01), while the level of TNF-α, IL-1β, TNF-α/IL-10 ratio (4.55±0. 69 vs. 5.18±0.38), BNP, hsCRP and LHFQ were decreased (P＜0.05 or P＜0.01) in CHF treatment group. Conclusions Thymopetidum, as an immunemodulating agent, might regulate the equilibrium of cytokines and improve the heart function of patients with CHF, indicating that immune modulation therapy might improve the treatment strategy for CHF patients.

Objective To investigate the changes of extraceUular regulated kinase 1/2 (ERK1/2) phosphorylation and assess the effects of blocking the ERK1/2 phosphorylation on rats with acute necrotizing pancreatitis (ANP). Methods The ANP model was induced by retrograde injection of 5% sodium tanrocholate into the biliary and pancreatic duct. 5 rats were treated as normal control. Other 75 Sprague-Dawley (SD) rots were randomly divided into sham operations(SO) group (n =25), ANP group (n =25) and PD98059 group (n =25). The rats were sacrificed at 15 min, 30 min, 1 h, 3 h and 6 h after ANP induction, the blood and pancreatic sample were taken. Pathological changes of pancreas were observed with light microscope and scored. The serum level of TNF-α and IL-1β was determined by ELISA. MPO activities in pancreas were measured by enzyme chemistry assay. Western blotting was performed to determine the phosphorylations of ERK1/2 in the pancreas homogenates. Results There was no significant pathologic changes in rats of SO group;but significant injuries occurred in ANP group, the pathologic score at 3 h was 9.9 ± 0.4;the extent of injuries attenuated in PD98059 group, the pathologic score at 3 h was 4.0 ± 0.4 (P < 0.05). The serum levels of TNF-α at 3 h in SO, ANP and PD98059 groups were (65.8 ± 20.5) pg/ml, (286.5 ± 50.3) pg/ml, (180.4±32.9)pg/ml, respectively;the serum levels of IL-1β at 3 h in SO, ANP and PD98059groups were (85.8 ± 25.5) pg/ml, (293.8 ± 46.3) pg/ml, (200. 5 ± 33.6) pg/ml, respectively;MPOactivities in pancreas were (0. 19 ± 0.02)U/g, (0.61±0.05)U/g, (0.52±0.03) U/g, and the values in ANP and PD98059 groups were significantly higher than those in SO group, while the values in PD98059 group were significantly lower than those in ANP group (P < 0.01). The expression of ERK1/2 phosphorylation in normal pancreas was 1100 ± 141, the expressions of ERK1/2 phosphorylation in ANP group at 15 min, 30 min were 5300 ± 486, 5621 ± 384, respectively;the expressions began to decrease 1 h later and returned the similar level as SO group at 6 h;the expressions of ERK1/2 phosphorylation in PD98059 group at 15 min, 30 rain were 4200 ± 370, 3600 ± 290, respectively;which were signifieanfly lower than those in ANP group (all P value < 0. 01). Conclusions The ERK1/2 signal transduetion pathway plays an important role in the pathogenesis of ANP. Inhibition of ERK1/2 phosphorylation by PD98059 may decrease the production of IL-1β, TNF-α and pancreatic MPO, attenuate the extent of pancreatic pathologic injuries.

Objective To explore the role of EphB4 in proliferation of glioma cells. Methods The mRNA and protein expressions of EphB4 were detected using RT-PCR, immunochemistry, and Western-blot, respectively. EphB4 siRNA was synthesized and transfected into U251 cells using Lipofectamine 2000. Glioma cell proliferation, apoptosis, and invasion were determined by MTT assay, TUNEL and transwell experiment, respectively. Results The expression (P<0.05) and proliferation of EphB4 were obviously decreased in U251 transfected with EphB4 siRNA and the proliferation was further decreased with the increased concetrations of siRNA. Compared with U251 group and siRNASCR group, EphB4 siRNA at different concentrations (25, 50 or 100 nmol/L) significantly reduced the invasion ability of cells and increased the number of apoptotic cells (P<0.05). Conclusions EphB4 plays an important role in the regulation of glioma cell proliferation, apoptosis and invasion.

This study was aimed to optimize the near infrared (NIR) variable selection method based on multivariate detection limit (MDL). Using Qing-Kai-Ling (QKL) injection as object, three variable selection methods (interval par-tial least-squares, iPLS; backward interval partial least squares, BiPLS; moving window interval partial least squares, mwPLS) were used to establish the PLS models of baicalin in QKL injection, respectively. The prediction ability of different variable selection method was compared. MDL of all models were calculated in contrast to the MDL value of full spectra PLS model, to select optimal variable selection method. The results showed that different variable selec-tion methods had different prediction ability. Among them, iPLS had the best performance which determination coef-ficient of prediction (Rpre2) and the root mean square errors of prediction (SEP) were 0.996 5 and 602.3 μg·mL-1, re-spectively. All MDLs of different variable selection methods were reduced compared with the full spectra PLS model. The value of iPLS was the lowest comes to be 1.19 μg·mL-1. The results above indicated that the best variable se-lection method for baicalin in QKL injection was iPLS. MDL theory took the error of calibration and validation set and the leverage of external sample into account, which can comprehensively evaluate model detection performance compared to the classic chemical indicator parameters. This method was particularly suitable for the variable selec-tion method optimization of NIR quantitative model of low concentration sample such as Chinese herbal medicine.