DTC is the most common endocrine carcinoma and its routine treatment consists of total thyroidectomy and 131I thyroid remnant ablation.Currently,standard follow-up for DTC comprises Tg measurement and neck ultrasound as well as an additional radioiodine scan when indicated.As thyroid cells are assumed to be the only source of Tg in serum,circulating Tg serves as an excellent marker of persistent or recurrent disease in DTC follow-up.With the development of highly sensitive Tg assays,now it is possible to detect very low Tg concentrations which reflect minimal amounts of thyroid tissue without the need for TSH stimulation.This review is to introduce clinical implications of highly sensitive Tg assays.

Objective To investigate the effects of microRNA (miR)106a on the proliferation, apoptosis, migration and invasion of thyroid cancer cells in vitro.Methods 8505C and CGTH-W3 cell lines were used in the study.Overexpression and inhibition of miR106a were achieved by transfection of lentiviral vectors.The changes of gene expression were detected by quantitative real-time PCR (qRT-PCR) and Western blot analysis.Cell viability and apoptosis were evaluated by MTT assay and flow cytometry analysis, respectively.The caspase-9 activities in parental CGTH-W3 and 8505C cells and transfected sublines were measured.Wound healing and Transwell invasion assays were performed to determine cell migration and invasion.Two-sample t test and one-way analysis of variance were used to analyze the data.Results The level of miR106a in 8505C was up-regulated when compared to that in CGTH-W3 cells (t=10.28, P<0.01).Scrambled control and miR106a(-) were also successfully transfected into cells.Inhibition of miR106a suppressed cell viability, migration and invasion while promoted apoptosis and caspase-9 activity of 8505C cells, with significant differences among 8505C, 8505C-control, 8505C-miR106a(-) cells (F=147.0, 19.2, 100.3, 537.8, 804.3;all P<0.01).Overexpression of miR106a promoted cell viability, migration and invasion while inhibited apoptosis and caspase-9 activity of CGTH-W3 cells, with significant differences among CGTH-W3, CGTH-W3-control, CGTH-W3-miR106a(+) cells(F=9.2, 13.3, 622.8, 12.3, 19.6, all P<0.01).In addition, miR106a may up-regulate the expression of MEKK2 and p-ERK1/2.Conclusion Acting as an onco-miR, miR106a might promote the proliferation, migration and invasion of thyroid cancer cells and inhibit their apoptosis in vitro.

Objective To explore the effect and mechanism of L-arginine on fetal growth restriction by observing the expression of Bcl-2 and Bax in placenta.Methods Sixty patients with FGR were chosen,among which 30 cases who were treated with conventional ways were as convention group,and the other 30 cases who were treated with L-arginine were as L-arginine group.The birth weight and perinatal fetus outcome were detected.The central tissue of placenta got within 10 min after delivery were fixed by 100 g/L formaldehyde and embed by pa-raffin wax to observe the expression of Bcl-2 and Bax using immunohistochemistry.SPSS 11.5 software was used to analyze the data.Results Compared with convention group,the birth weight of L-arginine group was higher(P

OBJECTIVETo construct recombinant lentivirus that stably suppresses phospholipase C (PLC) gamma1 expression in human colorectal carcinoma LoVo cells to obtain LoVo cell lines deficient in PLC gamma1 for investigation of the role of PLC gamma1 gene.

METHODSRecombinant lentivirus producing PLC gamma1 siRNA were constructed to infect LoVo cells, and the stably transduced cells were selected with blasticidin. The protein and mRNA expression of PLC gamma1 was examined by Western blotting and RT-PCR, and the effect of the lentivirus on cell apoptosis was analyzed by flow cytometry.

RESULTS AND CONCLUSIONPLC gamma1 siRNA significantly suppressed PLC gamma1 expression in LoVo cells, suggesting high efficiency of gene silencing induced by the siRNA produced by the recombinant lentivirus. Concomitantly, cell apoptosis induced by 5-FU was significantly increased.

Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; pathology ; DNA, Recombinant ; genetics ; Fluorouracil ; pharmacology ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Phospholipase C gamma ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective:To explore the expression of indoleamine 2, 3-dioxygenase 1(IDO-1), lymphocyte-activation gene 3 (LAG-3) and T cell immunoglobulin domain and mucin domain-containing molecule 3 (TIM-3) in differentiated thyroid cancer (DTC), and the value of them on prognosis.Methods:From May 2014 to November 2015, 119 DTC patients (33 males, 86 females, media age: 42 years) who underwent surgical treatment in Shanghai Sixth People′s Hospital were retrospectively analyzed. The expressions of IDO-1, LAG-3 and TIM-3 in the specimens were analyzed by immunohistochemistry and the expression differences between cancer tissues and normal tissues were analyzed by χ2 test. The correlation of IDO-1, LAG-3 and TIM-3 with clinical characteristics were analyzed using logistic regression analysis. The patients were followed up for 5 years, and the relationships of the progression-free survival (PFS) rate with the expressions of the three immune checkpoints were analyzed by Kaplan-Meier method, log-rank test and Cox proportional hazard models. Results:The overall 5-year PFS rate for 119 DTC patients (median follow-up time: 55(2-66) months) was 76.47%(91/119). The positive expression rates of LAG-3 and TIM-3 in cancer tissues were 21.85%(26/119) and 78.15%(93/119) respectively, which were significantly higher than those in normal thyroid tissues (7.34%(8/109) and 62.39%(68/109); χ2 values: 9.43, 6.81, both P<0.05). While the positive expression rate of IDO-1 was 70.59%(84/119) in cancer tissues, which did not show a significant difference from that in normal thyroid tissues (64.22%(70/109); χ2=1.05, P>0.05). Factors associated with the positive expression of LAG-3 included tumors with a single lesion (odds ratio ( OR)=0.248, 95% CI: 0.086-0.716, P=0.010). Log-rank test ( χ2=4.96, P=0.026) and multivariate Cox regression analysis (hazard ratio ( HR)=2.239, 95% CI: 1.013-4.592, P=0.046) suggested that LAG-3 positive expression was an independent risk factor of PFS. The same analysis of TIM-3 found no clinicopathological factors related to TIM-3 positive expression ( OR: 0.309-3.084, all P>0.05) and no association between TIM-3 positive expression and PFS ( χ2=0.008, P=0.929). Conclusion:The expressions of LAG-3 and TIM-3 are significantly increased in DTC tissues, and the higher expression of LAG-3 is associated with the worse prognosis, suggesting that LAG-3 may be a potential target for immunotherapy in DTC patients.

Objective To develop an effective way to evaluate the accurate platelet count in a patient with anticoagulants-induced pseudothrombocytopenia (PTCP).Methods It was studied that various anticoagulants effect on the platelets count for an infrequent patient with anticoagulants-dependent PTCP. When vitamin B6,aminophylline,gentamicin and amikacin were separately added to four anticoagulated blood samples from anticoagulants-dependent patient within 15 min after blood withdrawal,platelets count and morphological changes of blood cells after 4 hours of incubation at room temperature were investigated. The best anti-aggregating agent and its optimal concentration among them were explored.Results The four anticoagulants all could not inhibit the aggregation of the patient's platelets.Only amikaein among the above anti-aggregating agents can prevent and dissociate the aggregation of platelets without apparent morphological changes of blood cells and the platelet counts was stable within 4 hours after blood drawn when amikacin was added either before or after blood sampling.With increasing the concentration of amikaein,the platelet counts increase and then tend to be stable.The optimal concentration of amikacin is 5 mg/ml blood.Conclusions The supplementation of amikaein either before or after blood sampling is a useful method for the diagnosis anticoagulants-dependent PTCP and for the eva/uation of platelet counts in infrequent patients with anticoagulants-dependent PTCP.

Objective To establish and optimize three-cube FRET assay in living cells and analyze subunit assembling of iGluR receptors. Methods Taking HEK293 cells cotransfed with pECFP and pEYFP as negative control, and those transfected with pECFP-YFP as positive control,different calculation methods using fluorescence microscopy were compared. Results These calculation methods were all suitable for FRET measurement in the system. but the measurement results were affected by the ratio of Donor/Acceptor (D/A) in some degree,and different calculation methods have different optimized conditions. FRET measurement using FR value showed subunit specific assembly of iGluR subtypes.Conclusion There are different optimized conditions for these different calculation methods in the three-cube FRET measurement system,and a further evidence is provided for subunit specific assembling of iGluR subtypes from the FRET assay.

Objective To investigate the effect of olmesartan medoxomil on osteoprotegerin and serum inflammatory factor expression in atherosclerosis model rat.Methods From 2015 April in Jiangsu Institute of schistosomiasis control,24 Sprague-Dawley rats were randomly divided into the following three groups:the control group was fed with standard food,the model group was fed with high fat diet based on standard food(normal food +2％ cholesterol +5％ goat fat +0.2％ acid +7×105 IU/(kg·d) Vitamin D3),and the olmesartan group was given 3 mg/(kg·d) olmesartan gavage daily on the basis of high fat diet.On the fourth week of experiment the level of serum high sensitivity C reactive protein and serum lipids include total cholesterol,low density lipoprotein,high density lipoprotein and triglyceride were detected.The structure and changes of aortic pathology was observed.The expression of osteoprotegerinin in aortic was detected by immunohistochemistry staining and Western blot.Results Within the model group,the level of serum lipid and high-sensitivity C reactive protein increased significantly,endometrial thickness,intima-to-media thickness ratio and osteoprotegerin expression in aorta was significantly higher than that of normal group,the difference was statistically significant(P<0.05).The serum high density lipoprotein increased significantly,the level of other serum lipids and high sensitive C reaction protein decreased significantly,endometrial thickness,intima-to-media thickness ratio and osteoprotegerin expression in aorta of the olmesartan treated rats were significantly lower than those of model animals,the difference was statistically significant(P<0.05).Conclusion Olmesartan may suppress the development of atherosclerosis in model rats by decreasing the expression of osteoprotegerin and the level of serum inflammatory cytokines.

Objective To investigate the mechanisms by which nuclear factor-κB (NF-κB)translocates to nucleus of human glioblastoma U-87MG cells. Methods The levels of NF-κB translocating to nucleus in U-87MG after activation or inhibition ofphospholipase C-γ1 (PLCγ1) were first determined by electrophoretic mobility shift assay (EMSA), and then the levels of NF-κBtranslocating to nucleus in U-87MG after activation or inhibition of protein kinase-α (PKCα) were determined by EMSA. Results The level of NF- κB translocating to nucleus of U-87MG peaked 60min after EGF stimulation, which could be reversed by the pretreatment of PLCγ1 specific inhibitor U-73122. Moreover, PKC specific agonist phorbol myristate acetate (PMA) stimulation for 60 min promoted the nuclear translocation of NF-κB to the peak, which could be also effectively reversed by the pre-treatment of PKCα specific antagonist Ro 31-8220. Conclusions Epidermal growth factor can mediate the nuclear translocation of NF-κB in U-87MG through the signal transduction of PLCγ1-PKCα,thereby regulating the transcription of related invasion and metastasis genes.

OBJECTIVETo observe the ultracytochemical localization of H(+)-adenosine triphosphatase (H(+)-ATPase) in the cell organelles.

METHODSThe localization of H(+)-ATPase in the cell organelles was observed in the hepatocytes and renal cells of Wistar rats using routine ultracytochemical methods.

RESULTSH(+)-ATPase activities were observed on the lysosomal membrane and nuclear envelope of the hepatocytes and proximal tubule epithelial cells of the nephron in Wistar rats.

CONCLUSIONThis finding supports the hypothesis that H(+)-ATPase (V-ATPase) is present on the plasma membrane and in the endomembrane system.

Animals ; Cell Membrane ; enzymology ; Hepatocytes ; cytology ; enzymology ; ultrastructure ; Histocytochemistry ; methods ; Kidney ; cytology ; enzymology ; ultrastructure ; Lysosomes ; enzymology ; Male ; Organelles ; enzymology ; Rats ; Rats, Wistar ; Vacuolar Proton-Translocating ATPases ; metabolism