Objective To study the relationship between the lipoprotein associated phospholipase A2 (LP-PLA2) with the carotid plaques cerebral infarction,and study the predictive value of LP-PLA2 in carotid artery plaque stability.Methods According to the results of color doppler ultrasound examination of carotid artery,169 patients with cerebral infarction were random divided into cerebral infarction with carotid plaques group (101 patients) and cerebral infarction without carotid plaques group(68 patients) groups.According to the nature of plaque stability of carotid plaques.101 cases of cerebral infarction with carotid plaques group was divided into plaques group 30 cases and 71 cases of unstable plaque group.Set healthy control groups at the same time.Then detected level of LP-PLA2 for each patient by the method of double antibody sandwich enzyme-linked immunosorbent (ELISA).To evaluate the predictive value of LP-PLA2 in carotid artery plaque stability by mapping the receiver-operating characteristic (ROC) curve.Results The level of LP-PLA2 (212.90± 117.69 ng/ml) in carotid plaques group were significantly higher than those without plaque group (127.70 ± 57.96 ng/ml,t=3.016,P ＜0.01).It was not show significantly difference between no plaque group and healthy control group (108.34 ± 42.58 ng/ml,t=0.779,P＞0.05).But it showed significantly different between the unstable plaque group (236.24 ± 128.33 ng/ml)and stability plaques group (157.65±59.27 ng/ml,t=3.442,P＜0.01).Conclusion The LP-PLA2 of plasma could be involved in the development of atherosclerosis plaques.The LP-PLA2 can certain correlation with cerebral infarction of carotid plaques,can well evaluate the stability of carotid plaques.

Objective:To study the diagnostic value of combined detection of IAA,ICA and GADA in the classification of diabetes mellitus.Methods:30 cases of patients with type 1 diabetes who were treated in our hospital from June 2015 to June 2016 were selected as A group,60 cases of patients with type 2 diabetes were selected as B group,50 cases of healthy people were selected as C group.The IAA,ICA and GADA of the three groups were detected by ELISA,and the positive rate of the three groups were compared.Results:The fasting glucose of A group was (10.12± 3.68) mmol/L,B group was (11.23± 3.26) mmol/L,A group and B group were significantly higher than that of C group (P＜0.05),but there was no significant difference between A group and B group (P＞0.05);the positive rates of GADA,ICA and IAA in A group and B group were significantly higher than those in C group (P＜0.05),and the positive rates of GADA,ICA and IAA in A group were significantly higher than those in B group (P＜0.05);the sensitivity and specificity of combined detection of IAA,ICA and GADA in type 2 and type 1 diabetes mellitus were significantly higher than that in the single test (P＜0.05).Conclusions:The combined detection of IAA,ICA and GADA has a high diagnostic value in the classification of diabetes mellitus,which is worth clinical application.

Objective To discuss the aged people N-terminal pro-B-type natriuretic peptide (NT-proBNP)changes in thyroid function disorder and its correlation.Methods With electrochemical luminescence analyzer,detected serum NT-proBNP lev-el 38 patients with hyperthyroid heart disease,68 patients with hyperthyroidism,31 patients with hypothyroidism,and 43 ca-ses of healthy controls.Compared each groupserum NT-proBNP level of older population with middle-aged people,and anal-ysied the correlation.Results The serum NT-proBNP level of Hyperthyroidism heart group,hyperthyroidism,hypothyroid-ism group and normal control group were 827.61±626.13,107.18±54.46,162.94±134.14,68.76±39.21 pg/ml,respec-tively.The serum level of HT-pro BNP.Hyperthyroidism group,hyperthyroidism,hypothyroidism group compared with nor-mal control group,there were statistically significan(t = 7.458,4.312,3.794,P = 0.000,0.000,0.001).Hyperthyroidism heart,hypothyroidism with hyperthyroidism group serum level of NT-proBNP comparison there was statistical significance(t=7.078,2.232;P = 0.000,0.032).Hyperthyroidism heartgroup,hypothyroidism group and normal control group older population was higher than the level of serum NT-proBNP middle-aged,difference was statistically significant (t=-3.216,-2.510,-2.653;P =0.007,0.016,0.014).Hyperthyroidism group of elderly serum NT-proBNP level higher than that of middle-aged people,but there was no statistically significant difference (t=-0.140,P =0.890).Multiple regression analy-sis in hyperthyroidism group serum levels of NT-proBNP and FT4 had positive correlation (r=0.224,P =0.033)and hypo-thyroidismgroup serum levels of NT-proBNP and T3 had negative correlation (r=-0.363,P =0.022).Conclusion Thy-roid dysfunction in elderly people for the level of serum NT-proBNP had significant influence.Auxiliary disgnosis and cura-tive effect observation of the serum level of NT-proBNP in people with different thyroid functional status has certain clinical value.

Objective To analyze the sensitivity of clinical isolates of Candida albicans to flucona- zole,to detect mutations in their ERG11 genes,and to investigate the correlation between ERG11 gene mutation and resistance to fluconazole.Methods Candida albicans was identified from clinical isolates of Candida spp..The sensitivity to fluconazole was detected in vitro by microdilution-basesd method and Rosco tablets method.Three pairs of primers were designed to amplify three fragments of ERG11 gene(483 bps, from 295 bp to777 bp;482bps,from 723 bp to 1204 bp;489 bps,from 1179 bp to 1667 bp)after the extracting of genomic DNA.PCR products were sequenced.Results Eighty clinical isolates of Candida spp.were collected,which included 52 isolates of Candida albicans,all of which were sensitive to flucona- zole.Nineteen mutations were detected in ERG11 gene of 5 fluconazole-sensitive clinical isolates.Of the 19 mutations,14 were samesense mutations,and the remaing 5 missense mutations(T495A,A530C, G640A,A945C and G1609A),resulting in amino acid substitution D116E,K128T,E165K,E266D and V488I,respectively in lanosterol 14 alpha-demethylase.E165K was a novel mutation.Conclusions The clinical isolates of Candida albicans were highly sensitive to fluconazole;E165K and V488I might not lead to the resistance of Candida albicans to fluconazole.

UNLABELLEDIn this study, the roles of hematopoietic inhibitors elaborated by bone marrow endothelial cells in the proliferation and differentiation of hematopoietic progenitors were investigated. Murine bone marrow endothelial cell conditioned medium (BMEC-CM) was collected and the components with > 10 kD and < 10 kD were obtained by centrifugal ultrafiltration. The effect of BMEC-CM and its components on proliferation of hematopoietic progenitors was evaluated by CFU-GM and HPP-CFC assay and antibody neutralization test. The expression of the inhibitors in BMEC and BMEC-CM was detected by RT-PCR and Western blot, and change of proliferation and differentiation-related genes during expansion of hematopoietic progenitors was examined by membrane hybridization technique.

THE RESULTS(1) When BME C-CM and its components directly were added to CFU-GM and HPP-CFC culture system, BMEC-CM had no effect on colony formation, > 10 kD component enhanced and < 10 kD component inhibited the formation of CFU-GM and HPP-CFC. (2) When BMEC-C M and its components were added to liquid culture system of marrow cells, after 24 hours incubation, CFU-GM decreased and HPP-CFC increased significantly in B MEC-CM group, CFU-GM increased and HPP-CFC had no significant change in > 10 kD component group; and both CFU-GM and HPP-CFC reduced in < 10 kD group. (3) MIP-2, MIP-1 alpha, MSP, TGF-beta, TNF-alpha, IFN-gamma and T beta 4 were expressed in murine marrow endothelial cells, and MIP-2, MIP-1 alpha, MSP, TGF-beta, TNF-alpha and T beta 4 were existed in BMEC-CM. (4) Antibody neutralization test results demonstrated that TGF-beta, MSP, MIP-1 alpha, IFN-gamma and T beta 4 existed in BMEC-CM had significant suppressive effects on CFU-GM and HPP-CFC. (5) T beta 4 combined with 5 hematopoietic cytokines (SCF, IL-3, IL-6, GM-CSF and EPO) added to CD34(+) cells expansion culture system, HPP-CFC significantly increased compared with 5 cytokines group. T beta 4 could downregulated the expression of proliferation and differentiation-related genes and signal transduction-related genes. It is concluded that BM EC-CM promotes the proliferation of early hematopoietic progenitor cells, and this effect is related with the inhibitors existed in BMEC-CM and it could be executed via influencing cell proliferation and differentiation-related genes and signal-related genes.

Animals ; Bone Marrow Cells ; metabolism ; Cell Division ; drug effects ; Cytokines ; genetics ; pharmacology ; Endothelium ; cytology ; metabolism ; Hematopoietic Stem Cells ; drug effects ; physiology ; Mice ; RNA, Messenger ; analysis ; Thymosin ; pharmacology

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology

OBJECTIVETo investigate the toxicological mechanism of microcystin-LR (MCLR) on L-02 cells.

METHODSL-02 cells was treated with MCLR at different concentrations and the subsequent changes such as cell proliferation (MTT assay), morphology, lactate dehydrogenase (LDH) leakage, apoptosis rate and apoptosis-related gene expression were examined.

RESULTSMTT assay showed that MCLR mildly inhibited the cell growth within the initial 24 h of treatment but enhanced the cell viability after that till 60 h in a time- and dose-dependent manner. LDH leakage underwent no marked changes in response to 48-hour MCLR treatment but increased upon prolonged treatment for 60 h, indicating the presence of oxidative damage. After a 48-h treatment with MCLR at 50 microg/ml, obvious apoptosis of L-02 cells occurred as manifested by cell rounding, detachment from the substrate, cell shrinkage and membrane blebbing. The apoptosis rates were rather low (between 22% and 29%) after treatment with MCLR at different concentrations for 36 h, and increased to as much as 80% after a 60-h treatment with 50 microg/ml MCLR. The expressions of p53 and bcl-2 increased in the cells after treatment with high-concentration MCLR, suggesting that MCLR up-regulated the expression levels of the two proteins.

CONCLUSIONMCLR can induce apoptosis and up-regulate p53 and bcl-2 expressions in human normal liver cell line L-02.

Apoptosis ; drug effects ; Cell Line ; Hepatocytes ; cytology ; Humans ; Microcystins ; toxicity ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

Anoikis is a form of apoptosis induced upon cell detachment from extracellular matrix.It has been determined that acquisition of resistance to anoikis is a critical step for tumor cell metastasis.MiR-21,the most prominent oncomiR,plays an important role in tumor progression.In this study,we revealed that up-regulation of miR-21 in human esophageal adenocarcinoma (EA) is associated with lymph node metastasis and poor survival rate.Because of the established anti-apoptosis effect of miR-21,it is tempting to speculate that miR-21 might contribute to tumor metastasis by regulating anoikis,qRT-PCR analysis demonstrated that miR-21 expression in OE33/AR cells (subpopulation of human EA OE33 cells that acquired resistance to anoikis) was significantly increased.Also,transfection of miR-21 mimics provided OE33 cells resisting to anoikis.By luciferase assays,we verified that PDCD4 and PTEN were the functional targets of miR-21.In mouse model,via tail vein injection experiment,we showed that the metastasis formation of OE33 cells in vivo could be mediated by changing the miR-21 expression pattern.Taken together,our findings suggested that miR-21 was involved in the regulation of anoikis in human EA cells.Targeting miR-21 may provide a novel strategy to prevent metastasis.

To investigate whether lymphocyte function-associated antigen 1 (LFA-1) and very late antigen 4 (VLA-4) are involved in vasoendothelial adhesion and transendothelial migration of high proliferative potential endothelial progenitor cells (HPP-EPCs), flow cytometry was used to analyze the expression of integrin beta1 and beta2, the expression of intercellular adhesion molecule (ICAM-1, 2) and vascular cell adhesion molecule (VCAM-1) in mouse bone marrow endothelial cells (mBMECs). The adhesion and transmigration through endothelial cells of the HPP-EPCs blocked by functional grade neutralizing antibodies of VLA-4 and LFA-1 were studied in vitro. The results revealed that HPP-EPCs were positive for CD11a and CD49d in HPP-EPCs. The expression of ICAM-1and VCAM-1 of mBMECs increased after activated by IL-1beta and TNF-alpha. The results of adhesion in vitro revealed that the numbers of the adhered and migrated cells in the CD11a antibody group, in the CD49d antibody group and in the combinational antibody group were less than those in the isotype control antibody group. Furthermore, the number of adhered and migrated cells in the combinational antibody group was less than that in the CD11a or the CD49d antibody group (p < 0.05). It is concluded that both LFA-1 and VLA-4 are involved in vasoendothelial adhesion and transendothelial migration of HPP-EPCs.
Animals ; Antigens, CD ; metabolism ; Bone Marrow Cells ; cytology ; Cell Adhesion ; Cell Adhesion Molecules ; metabolism ; Cell Movement ; Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Integrin alpha4beta1 ; physiology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lymphocyte Function-Associated Antigen-1 ; physiology ; Mice ; Stem Cells ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo verify the clinical efficacy of heat-sensitive moxibustion in treatment of knee osteoarthritis (KOA).

METHODSSixty cases of KOA were randomly divided into a heat-sensitive moxibustion group and a conventional moxibustion group, 30 cases in each one. Dubi (ST 35), Yanglingquan (GB 34), Zusanli (ST 36) and Heding (EX-LE 2) on the affected side were selected in two groups. In heat-sensitive moxibustion group, the techniques of circling moxibustion, sparrow-pecking moxibustion, moving moxibustion and mild moxibustion were applied. In conventional moxibustion group, the mild moxibustion was used, 2 to 3 cm far from the skin of the acupoints selected. Lysholm scale for the assessment of knee joint function was adopted to evaluate the efficacy. The scores of joint pain, morning stiffness, joint swelling and walking ability were compared before and after treatment in two groups.

RESULTSThe scores of joint pain, morning stiffness, joint swelling and walking ability after treatment were all apparently improved as compared with those before treatment in either group (all P < 0.05). The improvement in the above-mentioned indices in heat-sensitive moxibustion group was much more apparent as compared with that in conventional moxibustion group (all P < 0.01). The effective rate was 90.0% (27/30) in heat-sensitive moxibustion group and was 73.3% (22/30) in conventional moxibustion group. The effective rate in heat-sensitive moxibustion group was obviously superior to that in conventional moxibustion group (P < 0.01).

CONCLUSIONThe efficacy of heat-sensitive moxibustion is superior to that of conventional moxibustion in the treatment of KOA. This therapy can more significantly improve the symptoms and physical signs of the patients with KOA.

Acupuncture Points ; Aged ; Female ; Humans ; Locomotion ; Male ; Middle Aged ; Moxibustion ; Osteoarthritis, Knee ; physiopathology ; therapy ; Treatment Outcome