BACKGROUND:Type 2 diabetic patients suffered from different levels of insulin resistance and insulin secretion defects. Retinol binding protein 4 is a fat derived factor found in the blood and is considered tobe an important target for the treatment of visceral obesity and insulin resistance.
OBJECTIVE:To observe the level of serum inflammatory cytokines and the changes of immune function in insulin resistance model rats after swimming.
METHODS:A total of28 8-week-old Sprague-Dawleymale ratmodels were randomly divided into swimming group and rest group. Rat received intraperitoneal injection with 3 μg/g recombinant serum retinol binding protein 4, twice a day, with an interval of 12 hours, for 6 weeks. At 6 weeks after administration, rats in the swimming group were subjected to swimming without loading for 6 weeks, 60 minutes per day. Serum retinol binding protein 4 levels were measured with enzyme linked immunosorbent assay (ELISA) kit. Insulin sensitivity (insulin resistance index) was assessed with homeostasis model assessment. Terminal deoxynucleotidyl transferase dUTP nick end labeling method was used to mark the apoptotic cels in rat thymus. ELISA was used to detect serum interleukin-6 expression. ABC-ELISA was used to test Level of serum granulocyte macrophage colony stimulating factor. S-P One Step Method was used to test CD4 and CD8 activities in blood.
RESULTS AND CONCLUSION:Interleukin-6 and granulocyte macrophage colony stimulating factor were significantly positively associated with insulin resistance index. The increase in insulin resistance index would induce the increase in the levels of serum inflammatory cytokine interleukin 6 and granulocyte macrophage colony stimulating factor, but exercises could decrease above levels. Thymus index, CD4, and CD4/CD8 were negatively associated with insulin resistance index. These results indicate that exercise intervention effectively improved immunity function of rats with insulin resistance and reduced insulin resistance index.

Objective To detect hepatitis C virus (HCV) RNA in amniotic fluid of gravida and investigate mother-to-infant transmission of HCV. Methods Thirty-four HCV seropositive gravida (experimental group) were engaged. Fluorescence quantitative polymerase chain reaction (PCR) based on amplisensor assay and reverse transcription -PCR (RT-nPCR) was used. Serum HCV RNA positive sera were genotyped by RFLP analysis of PCR products from 5′NC region. Sera and amniotic fluid samples of 40 normal gravida were set as the control group. Results In the experimental group, HCV RNA was detected in amniotic fluid (5.9%, 2/34) of 2 cases. HCV RNA titers were 10 5 and 10 6 copy/ml respectively. No HCV RNA was detected in the amniotic fluid and sera of the control (n=40). Conclusions HCV RNA was rarely detected in amniotic fluid. The amniotic fluid is not the main route of HCV mother-to-infant transmission.

Objective To construct the lentiviral vector encoding CLEC4M and prepare K -562 cells with stable overexpression of CLEC4M.Methods The gene sequence of normal CLEC4Mwas cloned by reverse transcription PCR and then inserted into GV166 vector to construct GV166-CLEC4Mlentiviral expression vector,and then lentiviral packaging was performed by transfection of293T cells.The ob-tained lentiviral liquid was used to infect human leukemia cell line K-562.Real-time PCR and Western blot were used to detect the over-expression of CLEC4M in K-562 cells.Results Sequencing showed that the recombinant lentiviral expression plasmid GV166-CLEC4M was successfully constructed.Lentiviruses could efficiently infect K-562 cells,according to real-time PCR.CLEC4Mwas successfully o-ver-expressed in K-562 cells at mRNA and protein levels.Conclusion The construction of lentiviral vector encoding CLEC4M lays a foundation for further study of CLEC4M gene involved in HCV entry into host cells.

Objective To evaluate the clinical features of central nervous system involvement in brucellosis,and to provide references for clinical treatment and diagnosis.Methods A retrospective review was carried out in 14 hospitalized patients who were diagnosed as neurobrucellosis from October 2008 to September 2011 at the First Hospital of Jilin University.Patients' clinical manifestation,pathogen characteristics,cerebrospinal fluid analysis and imaging data were recorded.Results Among the 14 cases of patients with neurobrucellosis,10 cases had the clinical manifestations of headache and vomiting,11 cases meningeal irritation,2 cases dysphoria and seizures,3 cases apathia,3 cases coma,3 cases positive pathological sign,and 2 cases lower extremities weakness.From the perspective of clinical diagnosis of 14 patients with neurobrucellosis,4 cases were meningitis,of which,meningitis with both lower extremities paralysis were 2 cases; encephalitis in 2 cases; meningoencephalitis in 6 cases; abducent paralysis,decreased vision,myelitis and ataxic gait in 1 case,respectively.All patients were in acute phase and had irregular fever.After combination therapy with antibiotics for 2 to 9 days,body temperature of all patients returned to normal,and they were discharged from the hospital after 1 to 2 weeks.All cases were followed up for 12 months,finding no sequelae.Conclusions Due to the diversity of central nervous system involvement in brucellosis,in case of a patient with long-term fever and neurological symptoms,medical and epidemiological history should be inquired and diagnosis would be made easy based on combination of cerebrospinal fluid with laboratory tests.Early treatment of acute neurological brucellosis has showed a good prognosis.

A RP-HPLC method for the quantification of the six major bovine milk proteins (κ-casein (CN),α_(s2)-CN,α_(s1)-CN,β-CN,Whey,immunoglobulins (Igg) ) is described. Separation and quantification were achieved by a reversed phase analytical column (Agilent Zorbax 300SB-C_8,250 mm×4.6 mm ,5μm) and the gradient elution solvents of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in acetonitrile at a flow rate of 0. 8 mL/min. Column temperature was set at 45℃ and the sample was monitored with photodiode array detector at 214 nm. A linear relationship( r >0. 999) between the concentrations of proteins and peak areas was observed over the concentration range. Recoveries of six target proteins spiked in milk were form 74. 8% to 132.5%. Nine kinds of milks of different brands were analyzed,and the difference of the concentration and relative ration of κ-CN,α_(s2)-CN,α_(s1)-CN,β-CN and Whey were found.

Objective To Study on anti-thrombosis effect of volatile oil of Acorus Tatarinowii Schott(AST) and ?-asarone.Methods Thrombosis test,blood hyperviscocity test,blood clottingtest,and in vitro fibrinolytic test were carried out to investigate the effects of volatile oil of AST and ?-asarone on the following indexes:weight of thrombus,prothrombin time(PT),activated part prothrombin time(APTT),hemorheology,clotting time,the weight of plasma fibrin clot.Results Volatile oil of AST and ?-asarone can lower the weight of venout thrombosis in rats,and prolong PT and APTT of blood plasma in rats.They can improve hemorheology in the hyperviscocity rats,especially reduce viscosity of whole blood and blood plasma obviously.They can prolong the clotting time in mice obviously;and decrease the weight of plasma fibrin clot.Conclusion Volatile oil of SCP and ?-asarone has anti-thrombosis anticoagulationand fibrinolyitc effect,and its mechanism may be related to the prolongation of PT and APTT,improving blood viscosity.

Purpose　To study the effects of trihexyphenidyl (THP) on levels of monoamine neurotransmitters in the cerebral cortex after subarachnoid hemorrhage (SAH).　Methods　SAH model of rats were used,the levels of norepinephrine (NE),dopamine(DA),5-hydroxytrypatamine (5?HT) and hydroxyindoleacetic acid (5?HIAA) were measured by flurospectrophotometry.　Results　There was an extensive increase in levels of NE (P<0.01),5?HT (P<0.01) and 5?HIAA (P<0.01) in the cerebral cortex after SAH,DA had a tendency to increase without significance.The increase in levels of NE (P<0.01),5?HT (P<0.01) and 5?HIAA (P<0.05) in the cerebral cortex after SAH could be effectively inhibited by THP.　Conclusions　There will be a remarkable increase in levels of NE,5HT and 5HIAA in the cerebral cortex after SAH,THP could significantly ameliorate the metabolic disorder of NE and 5HT after SAH

A series of phthalazine ketone compounds were synthesized and the structures were confirmed by H NMR and HR-MS spectrum. All target compounds were obtained through 7 steps, including selective reduction, nitration, bromination, ring enlargement, reduction, Knoevenagel and acylated reaction. The compounds were evaluated for their immunosuppressive effects of T-cell proliferation and inhibitory activity of IMPDH type II in vitro, as well as their structure-activity relationship were assessed. Several compounds exhibited strong immunosuppressive properties, especially compounds 7f and 7h, with IC50 values of 0.093 micromol x L(-1) and 0.14 micromol x L(-1) respectively, which were superior to mycophenolic acid. The information obtained from the studies may be useful for further research on the immunosuppressive agents.

BACKGROUND:Standing bending, supine bending, lateral pressing, pronation compression films and traction radiography are indeed effective to predict the correction outcome of surgeries by Harrington's method and Lugue's method,however, they are discontented for the correction of three-dimensional pedicle screw of vertebral arch.OBJECTIVE: To evaluate the role of preoperative X-ray films of gravity-suspending traction, supine bending and fulcrum bending in patients undergoing spinal three-dimensional correction and fusion for adolescent idiopathic scoliosis (AIS).DESIGN: Contrast observation.SETTING: Department of Orthopaedics, the 211 Hospital of Chinese PLA (Military Orthopaedics Center).PARTICIPANTS: From January 2003 to December 2005, 63 cases aged 10-21 years were recruited from Department of Orthopaedics in the 211 Hospital of Chinese PLA, including 18 males and 45 females. They were diagnosed as AIS by clinical examinations and X-ray film, with Cobb angle≥40°. Informed consents were obtained from all the subjects.METHODS: All of the 63 AIS cases with 79 structural curves were studied by preoperative X-ray films of standing,gravity-suspending traction, supine bending and fulcrum bending, which were compared with standing X-ray films one week postoperatively.MAIN OUTCOME MEASURES: Cobb angles of preoperative fulcrum bending film and postoperative standing film.RESULTS: All of the 63 cases were involved in the result analysis. There was no significant difference in the Cobb angle between preoperative fulcrum bending X-ray film and postoperative standing X-ray film (P ＞ 0.05). Gravity-suspending traction and supine bending films were significantly different from postoperative standing film (P ＜ 0.05).CONCLUSION: The fulcrum bending X-ray films are superior to gravity-suspending traction and supine bending films in predicting the postoperative correction of AIS patients.

Objective To observe the changes in forkhead/winged helix transcription factor p3(Foxp3), regulatory T cells(Treg),retinoid-related orphan receptor gamma(RORγt)in rat model of chronic obstructive pulmonary disease(COPD). Methods Twenty Sprague-Dawley(SD)rats were randomly divided into normal control group and COPD model group,with 10 rats in each group. The COPD model was reproduced by smoke inhalation and tracheal instillation of lipopolysaccharide(LPS),and no such treatment was conducted in normal control group. Twenty-eight days after the model reproduction,the pulmonary function was determined,the pathological changes of lung tissue were observed with haematoxylin-eosin(HE)staining,interleukins(IL-6,IL-10)in serum were detected by enzyme-linked immunosorbent assay(ELISA),CD4+CD25+Foxp3+Treg of peripheral blood was determined by flow cytometry,and the expressions of Foxp3,RORγt,IL-17 protein in lung tissue were assayed by Western Blot. Results Under light microscope,significal interstitial infiltration of inflammatory cells was found in alveoli and interstitial tissue of the lung,and destruction of alveolar tissue,alveolar wall thinning,and even rupture to fuse into bullae,and bleeding into alveoli in different degress could be observed. Compared with the normal control group,forced vital capacity(FVC),forced expiratory volume in 0.3 second(FEV0.3),FEV0.3/FVC,peak expiratory flow(PEF)in model group were significantly decreased〔FVC(mL):8.04±2.03 vs. 9.97±2.14,FEV0.3(mL):6.16±2.23 vs. 8.84±2.12,FEV0.3/FVC:0.70±0.09 vs. 0.85±0.11,PEF(mL/s):33.56±4.76 vs. 40.14±5.64, P<0.05 or P<0.01〕. Serum IL-6 was obviously increased(ng/L:93.17±20.96 vs. 76.28±13.24,P<0.05), IL-10 was significantly decreased(ng/L:78.62±15.17 vs. 104.34±19.46,P<0.01),and CD4+CD25+FoxP3+Treg was significantly diminished〔(2.75±0.83)% vs.(4.16±1.14)%,P<0.01〕in model group compared with those in the normal control group. The expression of Foxp3 protein in lung tissue in model group was significantly down-regulated compared with that in the normal control group(gray scale:0.38±0.15 vs. 0.63±0.11,P<0.01), and RORγt and IL-17 protein expressions were significantly up-regulated〔RORγt(gray scale):0.96±0.23 vs. 0.47±0.11,IL-17(gray scale):1.02±0.24 vs. 0.34±0.08,both P<0.01〕. Correlation analysis showed that FEV0.3 was positively correlated with Foxp3(r=0.585,P<0.05),and FEV0.3/FVC was negatively correlated with IL-6 and RORγt(r=-0.655,r=-0.607,both P<0.05). PEF was positively correlated with Treg(r=0.573, P<0.05),and negatively correlated with IL-17(r=-0.198,P<0.05). IL-6 was negatively correlated with Foxp3(r=-0.603,P<0.05),and positively correlated with RORγt(r=0.588,P<0.05). IL-10 was positively correlated with Treg(r=0.573,P<0.05). Treg was positively correlated with Foxp3(r=0.607,P<0.05), and negatively correlated with IL-17(r=-0.569,P<0.05). Foxp3 was negatively correlated with RORγt(r=-0.591, P<0.05). RORγt was positively correlated with IL-17(r=0.578,P<0.05). Conclusion There is a relationship among decreased pulmonary function,inflammation and imbalance of Foxp3/Treg and RORγt/Th17 in COPD.