OBJECTIVETo study the correlation between arachidonic acid (AA) and acute red blood cells damage in rats, and to build a model with hidden blood loss in vivo, and to explore the pathological mechenism of hidden blood loss.

METHODSA total of 50 male adult Sprague-Dawley rats weighing (200 ± 20) g were randomly divided into five groups (n = 10): control group and four experimental groups. The rats in the experimental groups were given 0.5 ml different concentrations of AA dilu- ents, 5, 10, 20, 40 mmol/L respectively. The blood samples were collected from orbital venous at the beginning and 24, 48, 72 hours after administration. Then the changes of hemoglobin (Hb) ,red blood cell count (RBC), glutathione peroxidase (GSH- PX) activity, total superoxide dismutase (T-SOD) activity and hydrogen peroxide (H202) in the blood samples were tested.

RESULTSSignificant hidden blood loss occurred when the concentration was 10 mmol/L in the experimental group, with the RBC and Hb sharply reduced in blood samples. The Hb and RBC were reduced in all the experimental groups and control group at 24 hours after administration, while in the experimental groups they changed more obviously. The GSH-PX activity, T-SOD activity and H₂O₂were also significantly reduced in all groups, and the changes showed significant differences. The Hb and RBC were relatively stable in the control group and the experimental groups at 48 hours after administration; while GSH-PX activity, T-SOD activity and H₂O₂were all significantly decreased, and the changes in the experimental groups were more notable.

CONCLUSIONElevated levels of AA in the blood causes oxidative stress in the red blood cells, leading to the damage of red blood cells and hemoglobin, which is responsible for hidden blood loss.

Animals ; Arachidonic Acid ; toxicity ; Erythrocytes ; drug effects ; metabolism ; Glutathione Peroxidase ; blood ; Hemoglobins ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Objective: To measure serum testosterone level, plasma levels of interleukin-18 (IL-18) and IL-10 and explore their correlation in patients with different types of coronary heart disease (CHD), and their possible role in occurrence and development of CHD. Methods: A total of 96 male CHD patients were divided into acute myocardial infarction (AMI) group (n=35), unstable angina pectoris (UAP) group (n=32) and stable angina pectoris (SAP) group (n=29). Another 30 patients who were excluded for CHD by coronary angiography were enrolled as non CHD control group. Enzyme linked immunosorbent assay (ELISA) was used to measure serum testosterone level and plasma levels of IL-18 and IL-10 in all groups. Results: Compared with non CHD control group, there were significant decreases in serum testosterone level [(13.46±1.99) mmol/L vs. (6.89±1.35) mmol/L vs. (5.02±1.87) mmol/L, t=1.917～2.365, P<0.05 both] in UAP group and AMI group, and that of AMI group was significantly lower than that of UAP group, t=1.034, P<0.05; there were significant increases in IL-18 levels [(146.72±79.36) pg/ml vs. (209.32±80.49) pg/ml vs. (316.78±75.63) pg/ml vs. (457.78±83.21) pg/ml, t=2.016～3.167,P<0.05 all] in SAP group, UAP group and AMI group, and those of UAP group and AMI group were significantly higher than that of SAP group, t=2.173, 2.596, P<0.05; there were significant increases in IL-10 levels [(48.46±18.27) pg/ml vs. (116.45±42.76) pg/ml vs. (85.64±27.33) pg/ml vs. (70.26±18.55) pg/ml, t=2.997～2.018, P<0.05 all] in SAP group, UAP group and AMI group, and those of AMI group and UAP group were significantly lower than that of SAP group (t=2.034, 2.291, P<0.05 both). Pearson linear regression analysis indicated that serum testosterone level was negatively correlated with levels of IL-10 (r=-0.678, P<0.01) and IL-18 (r=-0.579, P<0.01) in CHD group. Conclusion: There are significant changes in serum testosterone level and plasma IL-18, IL-10 levels, and testosterone level is significantly negatively related with IL-18, IL-10 levels, and they can be regard as new indexes assessing coronary atherosclerotic lesion.

Objective To observe the changes of serum sphingosine-1-phosphate (S1P) level in asthmatic patients with different severity of bronchial asthma, and to explore the evaluation value of S1P on the severity of asthma.Methods A prospective observational study was conducted. Fifty-two patients with asthma admitted to Department of Respiratory Medicine of the First Affiliated Hospital of Jiamusi University from November 2015 to January 2017 were enrolled. According to the severity of the disease, the patients were divided into mild, moderate and severe groups. In the same period, 25 healthy subjects were served as healthy control group. All the subjects got the peripheral venous blood collection in the morning fasting, the level of serum S1P was determined by enzyme linked immunosorbent assay (ELISA), the peripheral blood eosinophil (EOS) was counted, and the pulmonary function test was performed. The correlation among the parameters was analyzed by Pearson correlation analysis. Receiver operating characteristic curve (ROC) was plotted, and the value of serum S1P on evaluating the severity of asthma was analyzed.Results Fifty-two asthma patients were enrolled, including 17 patients of the mild, 19 of the moderate, and 16 of the severe. Compared with the healthy control group, serum S1P level and peripheral blood EOS in different degree asthma groups were significantly increased, and forced expiratory volume in 1 second (FEV1) was decreased significantly; and with asthma exacerbations, serum S1P levels and peripheral blood EOS were gradually increased [mild, moderate and severe S1P (nmol/L) were 1537.0±120.3, 1980.7±149.5, 2202.2±117.2 (F= 274.624, P= 0.001); EOS (×109/L) were 0.13±0.06, 0.20±0.07, 0.37±0.14 , respectively (F= 44.093,P = 0.001)], and FEV1 was decreased gradually [mild, moderate and severe were 0.89±0.05, 0.63±0.06, 0.42±0.10, respectively (F= 159.756,P = 0.001)]. Correlation analysis showed that there were significant positive correlations between serum S1P level and peripheral blood EOS in patients with mild, moderate and severe asthma (r value was 0.696, 0.746,0.508, allP < 0.05), and negatively correlations with FEV1 were found (r value was -0.761, -0.655, -0.815, all P < 0.01). There was no significant correlation between serum S1P level and EOS, FEV1 in healthy control group (r value was 0.324 and -0.048, bothP> 0.05). ROC curve analysis showed that the area under curve (AUC) of serum S1P for assessing mild, moderate and severe asthma was 0.948, 1.000, 1.000, respectively; when the cut-off of S1P was 1181.8, 1534.2, 1708.6 nmol/L, the sensitivity was 88.2%, 100%, 100%, and the specificity was 88.0%, 100% and 100%, respectively.Conclusions During asthma attack, the serum S1P level was gradually increased with the exacerbation of the disease. Serum S1P level has significant evaluative effect on the severity of asthma.

Objective To explore the immunogenicity of recombinant chimeric 6Aβ15-T including the Aβ1-15 epitope and a T-helper epitope formulated with different adjuvants and to evaluate its feasibility as a candidate vaccine for Alzheimer disease (AD).Methods The recombinant chimeric antigen 6Aβ15-T formulated with Al adjuvant, Freund′s adjuvant or MF59 adjuvant was administered to two strains of mice .The 6Aβ15-T-immunized group without adjuvants ( Mock) and non-immunized group (Control) were included in this study as control groups .The specific antibody and cellular immune response of the chimeric antigen were evaluated .Results In BALB/c strain mice, three types of adjuvants could substan-tially boost the immunogenicity of chimeric antigen 6Aβ15-T and produce a high level of specific-Aβ(β-amyloid) antibod-ies.In C57BL/6 strain mice, the existence of adjuvants enhanced the immune response of 6Aβ15-T antigen, but the mice in Mock group also produced a strong antibody response .In two strains of mice, prevalence of anti-AβIgG1, which was an indicator of Th2 polarization, was observed in the 6Aβ15-T-immunized mice.Additionally, the Al adjuvant induced a high-er level of IgG1 antibody titers, and the ratio of IgG1/IgG2a was the largest.As expected, the 6Aβ15-T antigen formulated with or without adjuvants induced PADRE-specific, but not Aβ42-specific T cellular immune response .Conclusion The 6Aβ15-T antigens formulated with different types of adjuvants could induce strong Th 2-polarized Aβ42-specific antibody re-sponses without activating self-reactive Aβ42-specific T cells in two strains of mice .The results suggested that the recombi-nant chimeric antigen 6Aβ15-T is a good candidate vaccine for AD .

To ensure the safety of the commercially available chenpi, a convenient and fast analytical method was developed for the determination of 133 pesticide residues in chenpi using gas chromatography-tandem mass spectrometry (GC-MS/MS). In this study, different extraction solvents, redissolution solvents and adsorbents were tested according to the recovery and purification effect to obtain a modified QuEChERS method. The samples were extracted with acetonitrile. During the clean-up step, octadecyl-modified silica (C18) and graphitized carbon black (GCB) were selected, and aminopropyl (NH2) was used instead of primary secondary amine (PSA) because of its weaker ion exchange capacity which had little effect on the recovery of ditalimfos. Samples were quantified by matrix-matched calibration with internal stan-dards. All pesticides showed good linearity in the respective range, both with values of r2 >0.99. The average recoveries of the pesticides spiked samples ranged from 70.0％ to 112.2％ with the RSDs of 0.2％–14.4％. The modified QuEChERS method was validated and applied to twenty real samples. Five pesticides were found in eight batches, but no pesticide exceeded the maximum residue limits (MRL, MRL reference to European commission).

BACKGROUND: During spinal orthopedic repair, the main difficulty is to maximize the correction of the deformity, simultaneously, to reduce the incidence of trauma and complications, especial y to avoid the corresponding spinal nerve injury. OBJECTIVE: To evaluate the curative effect of one-stage posterior pedicle subtraction osteotomy of apical vertebra with transpedicular instrumentation in the treatment of rigid angular kyphosis of thoracolumbar spine. METHODS: We retrospectively analyzed the data of 42 cases of rigid angular kyphosis of thoracolumbar spine that were treated by one-stage posterior pedicle subtraction osteotomy of apical vertebra with transpedicular instrumentation. Al patients received detailed imaging examination before and after operation. Kyphosis angle, Frankel grading and perioperative complications were recorded in al patients. X-ray films or CT films showed the bone graft fusion during fol ow-up. RESULTS AND CONCLUSION: Patients were fol owed up for 10-36 months after treatment. The average time of bone graft fusion was 5.1 months. Cobb’s angle of kyphosis was corrected from 78.4° (38°-110°) preoperatively to 7°(-8°-24°) at 10 days after treatment. The correction rate was 90%. The average angle during final fol ow-up was 7.9°, with an average loss of 0.9°. In 16 paresis patients, Frankel grading results showed grade B in 0 case, grade C in 3 cases, grade D in 5 cases, and grade E in 8 cases during final fol ow-up, showing significant improvement as compared with that pre-treatment (P < 0.05). Among 42 patients, 5 cases had complications. At 7 months after treatment, there were screw and titanium rod loosening at the distal end of the fusion segment in 1 case, cerebrospinal fluid leakage in 2 cases, transient double lower limb weakness in 1 case, pain in one side of lower limb in 1 case, and no severe complications appeared. These results verified that one-stage posterior pedicle subtraction osteotomy of apical vertebra with transpedicular instrumentation could achieve satisfactory clinical outcomes in rigid angular kyphosis of thoracolumbar spine. The internal fixation was stable and with a high fusion rate and few complications. The medium-term effect was satisfactory.

Objective To investigate the dynamic changes of dengue viral loads and IgM antibody in patients with dengue fever ( DF) and to analyze their relationships with disease severity. Methods A to-tal of 1 508 serum samples were collected from 1 140 hospitalized patients including 1 050 mild cases and 90 severe cases within 10 days after the onset of DF in Guangzhou in 2014. The viral loads were determined by using fluorescence quantitative RT-PCR. ELISA was performed to measure the dengue virus ( DENV)-spe-cific IgM antibody. Results In general, the DENV viral loads in patients declined gradually from 108 copies/ml on day 1 to 103 copies/ml on day 10 after the onset of DF. The viral loads in severe cases were significantly higher than those in mild cases on days 5 to 7 (P<0. 05). The positive rates of DENV RNA in serum samples also decreased with the disease progression from 100% on day 1 to 40% on day 10. Com-pared with the mild cases, the patients with severe DF showed higher positive rates of DENV RNA on day 6 and day 8 (P<0. 05). The DENV-specific IgM antibody could be detected on day 2 and the secretion of IgM antibody increased gradually with the disease progression. The levels of IgM antibody in mild cases were sig-nificantly higher than those in severe cases (P<0. 001). The positive rates of IgM antibody in patients in-creased form 8% on day 2 to 95% on day 6. Higher positive rates of IgM antibody were detected in mild ca-ses on days 5 and 6 as compared with those in patients with severe DF (P<0. 05). Conclusion High viral load and low level of IgM antibody during the fastigium of DF were closely associated with the disease severity.

ObjectiveTo study the correlation of urinary iodine and thyroid function in elderly men in Harbin,and to provide the basis for formulation of health measures for the elderly.MethodsSeventy five cases of clinically healthy elderly men were enrolled for check-up of urinary iodine,thyroid function and B-ultrasound in Geriatric Ward the Forth Affiliated Hospital of Harbin Medical University in 2010.The subjects of investigation were divided into iodine appropriate and iodine sufficient groups and thyroid function parameters and B-ultrasound results were compared.ResultsThe average age of the 75 cases of healthy elderly men was (79.07 ± 4.78) years old and the median of urinary iodine was 198.4 μg/L.There were 62.67％ (47/75) elderly males whose iodine nutritional status was appropriate,but there were still some individuals(6.67％,5/75) in the iodine excess state.The level of TSH of the iodine appropriate group [(1.91 ± 0.82)mU/L] was lower than that iodine sufficient group [(4.98 ±0.60)mU/L,t =12.58,P ＜ 0.05],while the level of FT3 of the iodine appropriate group[(4.71 ± 0.56)pmol/L]was higher than that iodine sufficient group[(3.31 ± 0.43)pmol/L,t =12.18,P ＜ 0.05].But the difference of FT4between the two groups [(14.91 ± 3.12),(14.06 ± 2.79)pmol/L] was not statistically significant (t =1.40,P ＞0.05].The thyroid volume of iodine sufficient group[(20.9 ± 6.1 )cm3] was higher than that iodine appropriate group [(17.9 ± 5.6)cm3,t =2.11,P ＜ 0.05].ConclusionsSufficient quantities of iodine intake may affect the thyroid of elderly people.Whether the quantity of iodine intake of the elderly population should be decreased or not need to be further studied.

Objective To evaluate the effect of progressive muscle relaxation combined with health education on the psychological state of the maintained hemodialysis patients and provide basis for the psychological intervention for them. Methods Forty-two MHD patients were randomly assigned to the intervention group and control group. Patients in the control group received the conventional nursing, and patients in the intervention group received the conventional therapy combined with nursing intervention for two times. Results After the first intervention, the various psychological testing indicators of intervention group were obviously lower than those of control group and there was statistically significant difference between two groups(P ＜ 0. 05 ). The differences of index of physiological chemistry between two groups had no statistical significance (P ＞ 0. 05 ). After the second intervention, the various psychological testing indicators of intervention group were further lower than those of control group and there was statistical significant difference ( P ＜ 0. 01 or P ＜ 0. 05 ). The Hb and Ca2 + of the intervention group were remarkably higher than control group,while PO43+ and K+ of intervention group were significantly lower than control group. Both differences had statistical significance(P ＜ 0. 05 ). Conclusions Progressive muscle relaxation combined with health education can greatly improve the psychological and physiological status of MHD patients.