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.

Purpose:To study the effect of early restrictive fluid resuscitation (EFR) on inflammatory and immune factors in patients with severe pelvic fracture (SPF).Methods:A total of 174 SPF patients in the Department of Orthopaedics,the First Affiliated Hospital of Chengdu Medical College from July 2015 to June 2018 were involved in this study and divided into EFR group (n =87) and control group (n =87) using the random number table method.Conventional fluid resuscitation (CFR) was performed in control group,and EFR was performed in EFR group.The incidences of acute respiratory distress syndrome (ARDS) and multiple organ dysfunction syndrome (MODS) during rescue,successful rescue rate,blood transfusion volume,fluid input,and resuscitation time were compared between the two groups.The parameters including prothrombin time (PT),hematocrit (HCT),platelet (PLT) and blood lactate (BL) at the 4th hour after fluid resuscitation were recorded.The levels of inflammatory factors (TNF-α,IL-6,CRP) and immune factors (CD3+,CD4+,CD8+,CD4+/CD8+) were compared between the two groups before treatment and 7 days after treatment.The revised acute physiologic and chronic health evaluation system and the sequential organ failure assessment scores were adopted for evaluation before treatment and 7 days after treatment.Results:The incidences of ARDS and MODS during rescue in EFR group were significantly lower than those in control group (p=0.015 and 0.010 respectively),and the successful rescue rate in EFR group was significantly higher than that in control group (p =0.011).The blood transfusion volume,fluid input,resuscitation time in EFR group were significantly lower than those in control group (p =0.016,0.002 and 0.001 respectively).At the 4th hour after fluid resuscitation,PTand BL in EFR group were significantly lower than those in control group (p =0.021 and 0.003 respectively),while HCT and PLT in EFR group were significantly higher than those in control group (p =0.016 and 0.021 respectively).On day 7 after treatment,TNF-α,IL-6,CRP and CD8+ in EFR group were significantly lower than those in control group (p =0.003,0.004,0.007 and 0.003 respectively),while CD3+,CD4+ and CD4+/CD8+ in EFR group were significantly higher than those in control group (p =0.004,0.000,0.007 respectively).On day 7 after treatment,the revised acute physiologic and chronic health evaluation (APACHE) system and the sequential organ failure assessment (SOFA) scores in EFR group were significantly lower than those in control group.Conclusion:EFR can effectively eliminate inflammatory factors,improve immune function,maintain the stability of blood components,reduce the incidences of ARDS and MODS,and elevate the successful rescue rate in patients with SPF.

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.

Objective To establish a colloid gold-immunochromatography assay (GICA) for detecting Plasmodiumfalciparum.Methods The monoclonal antibodies (mAbs) against lactate dehydrogenase ofP. falciparum (LDHpf) were screened for preparation of GICA strips. With microscopic examination of the blood smears and PCR test as control, GICAwas evaluated for its sensitivity, specificity and stability in the diagnosis of malaria in the outpatient clinics in China. Results Four hybridoma cell lines against LDHpf were prepared. Enzyme-linked immunosorbent assay demonstrated that the 4 mAbs reacted only with P. falciparum, but not with protein of normal human red cell, P. vivax, Toxoplasma gondii, or Schistosomajaponicam. All the 4mAbs recognized a 33 kD protein designated as LDHpf as shown by Western blot analysis. Compared with the microscopic examination of blood smears and PCR test, GICA had the sensitivities of 88.37% and 86.67% and the specificities of 95.65%and 97.78%, respectively. Concordance between microscopic examination and GICA for P. falciparum infection was91.55%.Conclusion GICA established in this study is simple, rapid, sensitive and specific for detecting P. falciparum, and is potentially useful in developing reagent kits for clinical use.

Objective To establish a colloid gold-immunochromatography assay (GICA) for detecting Plasmodiumfalciparum.Methods The monoclonal antibodies (mAbs) against lactate dehydrogenase ofP. falciparum (LDHpf) were screened for preparation of GICA strips. With microscopic examination of the blood smears and PCR test as control, GICAwas evaluated for its sensitivity, specificity and stability in the diagnosis of malaria in the outpatient clinics in China. Results Four hybridoma cell lines against LDHpf were prepared. Enzyme-linked immunosorbent assay demonstrated that the 4 mAbs reacted only with P. falciparum, but not with protein of normal human red cell, P. vivax, Toxoplasma gondii, or Schistosomajaponicam. All the 4mAbs recognized a 33 kD protein designated as LDHpf as shown by Western blot analysis. Compared with the microscopic examination of blood smears and PCR test, GICA had the sensitivities of 88.37% and 86.67% and the specificities of 95.65%and 97.78%, respectively. Concordance between microscopic examination and GICA for P. falciparum infection was91.55%.Conclusion GICA established in this study is simple, rapid, sensitive and specific for detecting P. falciparum, and is potentially useful in developing reagent kits for clinical use.

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