Recently,a large family of G-protein coupled receptors (GPCRs) has been identified as mas-related genes (Mrgs),Which are specific expressed in small diameter sensory neurons in the trigeminal and dorsal root ganglia,suggesting a role in pain transmission.Mrgs receptors have been shown to modulate some physiological and pathological activities,such as pain and immunity.It is important to discover ligands of Mrgs for understanding and elucidating their potential physiological and pathophysiological roles.The studies on pharmacological spectrum of NPS make it an interesting target for pharmaceutical development.

Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene,and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma.Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro,then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP.pLVX-CMV-0-EGFP was constructed as control.BMSCs were isolated,cultured,and transfected by lentivirus.The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected.The expression of NIS gene was examined by Western blot.Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4.The nude mice intracranial glioma models were established.MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein.Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged.BMSCs were successfully isolated and cultured.Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection.The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP,but not in BMSCs-EGFP.BMSCs-NIS-EGFP showed significantly more uptake of 125I (nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4.The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection.Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma.It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma.

A new medical research technology that combines surface enhanced Raman spectroscopy (SERS)with labeling immune technique is emerging with the development of SERS.This paper is intended to describe the principles, research progress and existing problems relating to SERS labeling immunoassay technology.We also summarize the research techniques for improving the sensitivity of SERS labeling immunoassay and the methods to eliminate nonspecific adsorption in SERS labeling immunoassay.Furthermore,the future development of SERS labeling immunoassay technology is discussed.

Objective To investigate the clinical significance and biological role of G-protein coupled receptor 49(GPR49) expression in pancreatic carcinoma.Methods GPR49 expression in tumor and adjacent normal tissues of 77 patients with pancreatic cancer was compared by tissue microarray and immunohistochemistry.And then, the GPR49 expression levels in the tumor tissues of patients with different pathological grades and clinical stages were analyzed.GPR49 positive pancreatic cancer cell line CFPAC-1 was taken as cellular model.CFPAC-1 cells were stimulated with roof plate-specific spondin(RSPO)1, the ligand of GPR49, in vitro.The effect of RSPO1 on CFPAC-1 cells proliferation was evaluated with cell counting.The effect of RSPO1 on the expression of membrane molecular CD44 in CFPAC-1 cells was detected by flow cytometry.CFPAC-1 cells incubated with RSPO1 were subcutaneously implanted into nude mice.And then, the time of tumor formation and tumor size were observed.T test and single factor analysis of variance were performed for statistical analysis.Results GPR49 was widely expressed in all 77 pancreatic cancer tissues.By immunohistochemistry, the score of GPR49 expression in pancreatic cancer tissues was 9.0±2.4, which was higher than that of adjacent normal tissues (5.7±2.4), and the difference was statistically significant (t=8.995, P<0.01).There was no correlation between GPR49 expression and tumor sizes, pathological grades, lymph node metastasis, and clinical stages (all P>0.05).The results of experiments in vitro indicated that RSPO1 could promote CFPAC-1 cells proliferation and up-regulate CD44 expression in CFPAC-1 cells.Experiments in vivo demonstrated that after 30 days the tumor volume of mice implanted with RSPO1-pretreated CFPAC-1 cells was (606.0±188.0) mm3, which was larger than that of PBS-pretreated group ((364.2±83.7) mm3), and the difference was statistically significant (t=2.616, P=0.031).Conclusion GPR49 is widely expressed in pancreatic cancer cells and RSPO1/GPR49 pathway has play a role in promoting the proliferation of pancreatic cancer cells, which might be a potential target for interfering pancreatic cancer.

Aim Toinvestigatetheeffectofbencyclo-quidium bromide(BCQB)on mucus MUC5AC expres-sion induced by lipopolysaccharide in cultured human nasalepithelialcells(HNECs).Methods Primary culture of human nasal epithelial cells (HNECs)was randomly divided into control group (C,with no treat-ment),LPS group (LPS,with LPS 1 mg · L-1 added in),BCQB low dose group(BCQBL,with LPS 1 mg· L-1 and BCQB 10 -8 mol·L-1 added in),BCQB mid-dle dose group(BCQBM,with LPS 1 mg·L-1 and BC-QB 10 -7 mol·L-1 added in),BCQB high dose group (BCQBH,with LPS 1 mg·L-1 and BCQB 10 -6 mol· L-1 added in)and ipratropium bromide group(IB,with LPS 1 mg·L-1 and IB 10 -6 mol·L-1 added in).Af-ter incubation at 37 ℃with 5% CO2 for 24 h,the ex-pression of MUC5 AC mRNA was detected with Real-time PCR and the expression of MUC5 AC protein in HNECs was detected with Western blot,while the ex-pression of MUC5 AC protein in supernatant was detec-tedwithELISAineachgroup.Results Ascompared with control group,the expression of MUC5 AC mRNA and protein increased significantly in LPS group (each P<0. 01 ).As compared with LPS group,the expres-sion of MUC5 AC mRNA and protein decreased signifi-cantly in each group of BCQB(P<0. 01,P<0. 05), and there was no statistical difference between BCQB high dose group and control group (each P>0. 05 ). Conclusion Bencycloquidiumbromidecansuppress MUC5 AC expression induced by LPS in cultured hu-man nasal epithelial cells,indicating that BCQB may be a new drug for nasal mucous hypersecretion diseases.

Objective To investigate the role of TL1A in the generation and differentiation of peripheral blood Th17 in rheumatoid arthritis.Methods The peripheral blood mononuclear cells (PBMCs) of RA were isolated and stimulated with PHA in the presence or absence of TL1A.Naive CD4+ T cells from RA was cultured under Th17-polarizing conditions with or without TL1A.The percentage of Th17 was detected by flow cytometry (FCM) analysis.The DR3 expression on CD4+ T cells from PBMCs of RA patients and healthy controls (HC) were analyzed by using FCM.Data were analyzed with t test and U test.Results TL1A could significantly up-regulate the percentage of Th17 in PBMCs of RA [(7.5±2.3)％ vs (5.2±1.5)％,t=2.647,P＜0.05].Compared to the HC groups,TL1A could significantly induce Th17 differentiation from naive T cells [(37.7±1.9)％ vs (29.5±2.0)％,t=6.455,P＜0.05].The percentage of CD4+DR3+ T cell in PBMCs of RA [(0.56±0.87)％] was significantly higher than that of HC [(0.13±0.04)％,P＜0.05].The percentage of CD4+DR3+ T cell in PBMCs when stimulated with PHA was increased in RA patients,[(4.51±1.34)％ vs (1.11±0.29)％,t=2.915,P＜0.05],but no obvious increase in HC [(0.199±0.104)％ vs 0.072％±0.029)％,t=1.644,P=0.1988].Conclusion TL1A can promote the generation and differentiation of Th17 in the PBMCs of RA,and this effect may be mediated by the binding of TL1A with DR3.

Objective To investigate the feasibility of differentiation between prostate cancer and prostatitis by using metabolic ratios provided by 3D 1H Magnetic Resonance Spectroscopic Imaging (MRSI). Methods Metabolic changes were evaluated in 42 voxels with prostate cancer and 30 voxels with prostatitis in the peripheral zone using MRSI. The results were based on the pathologic findings by biopsy. The (choline + creatine)/Citrate (CC/C) ratio and the changes of choline and citrate levels were evaluated in each voxel with cancer or prostatitis, t test was used to determine the power of the CC/C ratio in differentiation between prostate cancer and prostatitis. Results The CC/C ratio for cancer voxels (1.28±0.41) was significantly different from the ratio in the voxles with prostatitis (1.03±0.40), t=6.45, P<0.05, due to greatly increased choline level in the cancer voxels. When CC/C ratio of 0.8 was taken as the criteria for the diagnois of prostate cancer, the sensitivity, specificity and accuracy were 65.5%, 71.4% and 66.7%, respectively. Positive predictive value(PPV)and negative predictive value(NPV) were 90.5% and 33.3%, respectively. The CC/C ratio was higher than 0.86 in 66.7% voxels with prostatitis (20 voxels of total 30 voxels), which mostly depended on the level of choline. When citrate level was used as an auxiliary index to evaluate prostatitis (Cit/norm, Cit≥0.75), the misdiagnosis rate of prostate cancer was reduced to 26.6%(8 voxels of total 30 voxels). Conclusions The metabolic ratio of CC/C can be used to differentiate prostate cancer from prostatitis. The misdiagnosis rate is reduced when citrate is not or slightly decreased relative to normal citrate level (Cit/norm, Cit≥0.75).

Aged ; Biomarkers ; blood ; Cobalt ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; Myocardial Ischemia ; diagnosis ; Serum Albumin ; metabolism ; Spectrophotometry

Objective To evaluate tear ferning changes of conjunctivochalasis.Design Prospective case study series.Partici- pants 30 patients(60 eyes)of conjunctivochalasis and normal subjects were selected.Methods The subjects were observed with gen- eral ophthalmic examination and tear fern test(TFT).Tear ferning was classified into 4 types.TypeⅠand TypeⅡare normal.TypeⅢand TypeⅣare abnormal.Main Outcome Measures The type of tear feming.Results TFT showed that tear ferning was de- creased in conjunctivochalasis group(TypeⅢand TypeⅣoccupied 61.7%).The difference between conjunctivoehalasis and normal control group was significant(P

Based on information entropy theory, this study analyzes the experimental indicators of glucocorticoid-induced traditional Chinese medicine yang deficiency or yin deficiency syndrome in animal models, thus laying the foundation of defining and interpreting the model state.