The ionic liquid butylamine 2,5-dihydroxybenzoate (DHB-BuN) was chosen as the matrix for improving repeatability of oligosaccharides quantitation, and imaging oligosaccharides in soybean and leaf by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS).Using the ionic liquid matrix methanol solution (20%, V/V) to cover sample slice, and setting MS acquisition parameters including polarity as positive and laser power as 70%, the MS imaging experiment was performed to analyze oligosaccharides and their distribution in soyben and soybean leaf.As a result, the MALDI-MS analysis of 3 kinds of oligosaccharides including sucrose, raffinose and stachyose was performed using the ionic liquid matrix and the reproducibility RSDs were less than 3% for in-spot and less than 4% for spot-to-spot.The standard curves linear correlation coefficients (R2) were all no less than 0.996 between MS peak intensity and oligosaccharides concentraions in the concentration rage of 0.062-1.00 mg/mL, which showed quantitative power in MALDI-MS analysis.The matrix was used for MALDI-MS imaging analysis of soybean chips and soybean leaf surface disaccharide, trisaccharide and tetrasaccharide to obtain the soybean mass spectrum and spatial resolution of 150 μm of the three oligosaccharides imaging.Three kinds of oligosaccharides in the soybean roughly balanced distribution, and the oligosaccharides distribution in the leave was mostly at its tip part.The contents of the 3 kinds of oligosaccharides could be estimated by their MS peak intensity and above standard curve.The result suggested that the ionic liquid DHB-BuN matrix could be well applied in MALDI-MS imaging analysis of oligosaccharides in different sample.

Objective To observe the effect of hypertonic saline complex solution (hypertonic saline plus hydroxyethyl starch,HSH) on patients with severe cerebral trauma,high intracranial pressure and shock by the measurement of the changes of the mean arterial pressure (MAP),central venous pressure (CVP) and intracranial pressure (ICP),as well as GOS score changes followed up for 6 months,in order to determine the value of HSH treatment in severe cerebral trauma,intracranial hypertension and shock.Methods Sixty patients with severe brain injury and uncorrected hemorrhagic shock were selected,while the degree of coma was assessed by using GCS score,and shock severity was estimated by using the shock index (SI) score.The patients were randomly divided into HSH group (n =30) and mannitol group (MT group,n =30).Thirty minutes,60 min and 120 min after administration either solution,The changes of MAP,CVP and ICP were observed in two groups,and all patients were followed up for 6 months to observe the outcomes of patients.Results There were no statistically significant differences in age,gender,GCS score,SI scores,and other medication between two groups (P ＞ 0.05),and they were comparable between two groups.After resuscitation of patients in two groups,MAP and CVP were elevated,but the effect of HSH appeared sooner and higher within 30 minutes [MAP (63.1 ± 8.8) mmHg vs.(51.0-9.3) mmHg] (P ＜ 0.05);At the same time,ICP dropped more than 10％ lower [ICP (27.3 ± 5.9) mmHg vs.(32.8 ± 4.1) mmHg] (P ＜0.05),while the effect of MT appeared more slowly in hemodynamic improvement;at 120 min,the increase in MAP and reduction in ICP in HSH group were more significant than those in MT group [MAP (65.9 ± 13.2) mmHg vs.(60.4 ±7.2) mmHg] (P ＜0.01);the ICP [(22.2 ±4.7) mmHg vs.(28.1 ±6.1) mmHg] (P ＜ 0.01).Followed up for 6 months,good recovery rate in HSH group was higher and poor recovery rate was lower than those in MT group.Conclusions In patients with acute intracranial hypertension and uncorrected hemorrhagic shock,the employment of hypertonic saline plus hydroxyethyl starch solution can produce faster and more effective therapy for shock and reduce intracranial pressure,improving the long-term neurological function of patients.

BACKGROUND:Retinoid X receptor (RXR) plays a central role in the regulation of intracellular receptor signaling pathways. The activation of RXR has protective effect on H2O2-induced apoptosis of H9c2 ventricular cells in rats. But the protective effect and mechanism of activating RXR in cardiomyocytes against hypoxia/reoxygenation (H/R)-induced oxidative iniury are stillunclear. METHODS:The model of H/R injury was established through hypoxia for 2 hours and reoxygenation for 4 hours in H9c2 cardiomyocytes of rats. 9-cis-retinoic acid (9-cis RA) was obtained as an RXR agonist, and HX531 as an RXR antagonist. Cultured cardiomyocytes were randomly divided into four groups:sham group, H/R group, H/R+9-cis RA -pretreated group (100 nmol/L 9-cis RA), and H/R+9-cis RA+HX531-pretreated group (2.5 μmol/L HX531). The cellviability was measured by MTT, apoptosis rate of cardiomyocytes by flow cytometry analysis, and mitochondrial membrane potential (ΔΨm) by JC-1 fluorescent probe, and protein expressions of Bcl-2, Bax and cleaved caspase-9 with Western blotting. Allmeasurement data were expressed as mean±standard deviation, and analyzed using one-way ANOVA and the Dunnett test. Differences were considered significant whenP was <0.05. RESULTS:Pretreatment with RXR agonist enhanced cellviability, reduced apoptosis ratio, and stabled ΔΨm. Dot blotting experiments showed that under H/R stress conditions, Bcl-2 protein level decreased, while Bax and cleaved caspase-9 were increased. 9-cis RA administration before H/R stress prevented these effects, but the protective effects of activating RXR on cardiomyocytes against H/R induced oxidative injury were abolished when pretreated with RXR pan-antagonist HX531. CONCLUSION:The activation of RXR has protective effects against H/R injury in H9c2 cardiomyocytes of rats through attenuating signaling pathway of mitochondria apoptosis.

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BACKGROUND:Triggering receptor expressed on myeloid cel s 2 (TREM-2) is highly expressed throughout the synovial tissue in active rheumatoid arthritis patients, but the role of TREM-2 in the pathogenesis of rheumatoid arthritis stil remains unclear.
OBJECTIVE:To explore the TREM-2 expression in the synovial tissue of col agen type II-induced arthritis rats.
METHODS:The col agen-induced arthritis models were established in rats. The activity indicators and pathological changes of arthritis synovial were dynamical y observed. The mRNA levels of TREM-2, tumor necrosis factor-α, interleukin-1β, and interleukin-10 were detected in synovial tissue of rats by RT-PCR. The protein expression and location of TREM-2 were measured with western blot assay and immunohistochemistry, respectively.
RESULTS AND CONCLUSION:At day 13 after immunization, the paws of model rats appeared red and swel ing, the arthritis index scores were increased (P<0.01). At day 19-25 after immunization, the inflammation reached the peak. Hematoxylin-eosin staining showed that, the synovium of col agen-induced arthritis rats were proliferated and were infiltrated by inflammatory cel s, cartilage was destroyed. Compared with the control group, the expression of TREM-2 mRNA and protein, the mRNA levels of tumor necrosis factor-αand interleukin-1βin synovial tissue of the model rats were significantly increased (P<0.05 or P<0.01), while interleukin-10 mRNA expression was significantly decreased (P<0.05). Experimental findings indicate that, TREM-2 is a crucial inflammatory regulator and the increasing expression of TREM-2 plays an important role in the pathogenesis of col agen-induced arthritis.

AIM: To investigate the effects of transient receptor potential channel 6 (TRPC6) on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) induced by IL-1β.METHODS: The mRNA expression of TRPC6 in synovial tissues from RA or OA patients was studied by RT-qPCR.RA-FLS were cultured by enzyme digestion and tissue adhesion methods.The method of flow cytometry was applied to identify the RA-FLS.RA-FLS were treated with different concentrations (0, 0.25, 0.5, 1, 2, 4, 8 and 16 μg/L) of IL-1β for 36 h.The cell viability was examined by CCK-8 assay.RA-FLS were incubated with IL-1β (16 μg/L) for different time (12, 24, 36, 48, 60 and 72 h), and the cell viability was measured by CCK-8 assay.The interference efficiency of TRPC6-siRNA was determined by RT-qPCR and Western blotting.After incubation in the presence or absence of IL-1β medium, the cell viability, the percentage of EdU-positive cells and the percentage of (G2/M+S) phase were measured by CCK-8 assay, EdU labeling assay and flow cytometry, respectively.RESULTS: The mRNA expression of TRPC6 was found in synovial tissue with higher levels in RA patients than that in OA patients.TRPC6-siRNA significantly decreased the mRNA and protein expression of TRPC6 (P<0.05).When RA-FLS were treated with IL-1β, the proliferation of RA-FLS was increased (P<0.05).The differences of the cell viability, the percentage of EdU-positive cells and the (G2/M+S) phase percentage between TRPC6-siRNA group and blank control group or NC-siRNA group were significant, in the presence of IL-1β (P<0.05).However, they were not significant in the absence of IL-1β.CONCLUSION: TRPC6 is involved in the proliferation of RA-FLS induced by IL-1β.Silencing of TRPC6 gene inhibits the growth of RA-FLS induced by IL-1β.

ObjectiveTo explore the prevalence and clinical features of hyperuricemia and its influencing factors in elderly male people aged 90 years and above. MethodsOne hundred elderly male people aged 90 years and above who underwent routine health examination in our hospital in 2007 were selected in the study. Serum uric acid level was examined by uricase-peroxidase method, and all patients were divided into hyperuricemia group and control group according to the serum uric acid level. Clinical and biochemical indications were compared between the two groups, and logistic regression was used to analyze the influencing factors of hyperuricemia in elderly people. ResultsThe serum uric acid level was increased in 20% of the elderly people, and the prevalence of gouty arthritis was 1%. The levels of blood urea nitrogen and serum creatinine were higher in hyperuricemia group than in control group[(10. 98±4.29) mmol/L vs. (6. 87± 1.86) mmol/L, (125.2±25.9)μmol/L vs. (93. 4±19. 8)μmol/L;both P<0.05)3. The patients in hyperuricemia group had a higher prevalence of hypertension and hypertriglyceridemia, and a higher proportion of diuretic application than patients in control group(P<0. 05). Logistic regression analysis showed that serum uric acid level had the most remarkable correlation with serum creatinine(OR= 1. 969), followed by fasting blood glucose (OR= 1. 310)and blood urea nitrogen(OR = 1.161). There was negative correlation between serum uric acid level and plasma cholesterol level(OR = 0. 802). ConclusionsThe prevalence of hyperuricemia is high in elderly people aged 90 years and above, while the incidence of gouty arthritis is low. Renal function impairment, metabolic syndrome and thiazide diuretic are the major factors for hyperuricemia.

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To probe deficits in visuospatial attention using an attention network test (ANT) in patients with frontoparietal network lesions. Methods The ANT was used to measure the alertness, orienting and executive control abilities of 25 patients with local brain lesions, including 13 with frontal and 12 with parietal damage. Their results were compared with those of health adults. Results During ANT tasks, the patients' responses were significantly slower on each cue and target condition than controls', and showed deficits in their alerting and orienting networks. The efficiency of executive control was impaired in patients with frontal lesions, but increased with parietal lesions. Conclusions These findings suggest that the frontoparietal network is involved in alerting and orienting, but the executive control function may be selectively associated with the frontal lobe. ANT is an efficienttool for studying visual attention and cognition.

Objective To identify a novel pathogenicity mutation of acid alpha‐glucosidase(GAA) gene in a Chinese family with two siblings affected with juvenile onset form glycogen storage disease Ⅱ(GSD Ⅱ) .Methods The clinical and family data of two siblings presenting recurrent respiratory tract infections ,respiratory failure associated with systemic muscle weakness ,were an‐alyzed and diagnosed with GSD Ⅱ by detecting alpha‐1 ,4‐glucosidase activity .DNA was extracted from peripheral blood of the proband ,younger brother and his parents .All 20 exons and the intron‐exon splice sites of GAA gene were amplified by polymerase chain reaction (PCR) .Mutations were detected by direct sequencing the PCR products .Results The younger brother was found to be compound heterozygous for two mutations in the GAA gene :c .1216G>A (p .Asp406Asn) missense mutation in the exon 8 from his father and c .1935C>A (p .Asp645Glu) missense mutation in the exon 14 from his mother .Conclusion The compound hetero‐zygous c .1216G>A and c .1935C>A mutations caused the juvenile onset form GSD Ⅱ characterized by dyspnea and cardiac hyper‐trophy .The novel c .1216G>A mutation may be related to the juvenile onset form GSD Ⅱ .