Background The proliferation and migration of vascular endothelial cells is a primary link during angiogenesis.Studies showed that heat shock protein B6 (HspB6) promotes the secretion of multiple angiogenesis-related factors and therefore leads to neovascularization.Understanding the effects of neutralizing HspB6 antibody on the biological behavior of human choroidal vascular endothelial cells has an important significance in the target treatment of choroidal neovacularization diseases.Objective This study was to address the role and mechanism of neutralizing HspB6 antibody in tube formation of human choroidal vascular endothelial cells.Methods Human choroidal vascular endothelial cell line was normally cultured and harvested for total RNA extraction.Expressions of HspB6 mRNA and protein in human choroidal vascular endothelial cells were detected by reverse transcription PCR (RT-PCR) and flow cytometry (FCM).The cells were seeded on 96-well plate covered with matrigel at the density of 2×104/hole.Then the neutralizing HspB6 antibody at the concentration of 100 μg/Land 500 μg/L was added into the medium respectively,and the control cells were set without the addition of HspB6 antibody.The number of capillary tubes was calculated 12 hours after culture by three-dimensional matrigel assay.In addition,0,50,100,500 μg/L of neutralizing HspB6 antibody were added into the cell medium separately for 24hours,cell counting kit-8 (CCK-8) method was employed to assay the inhibitory rate(IR) of the cells.Transwell test was used to count the cell number across chamber membrane for the evaluation of migration ability of the cells.The apoptosis of the cells was assayed by FCM.Results Both HspB6 mRNA and protein were expressed on human choroidal vascular endothelial cells.The number of capillary tube formation of human choroidal vascular endothelial cells was (67.25±5.75),(60.39±6.41) and (39.76±10.73) /field in the 0,100 and 500 μg/L neutralizing HspB6 antibody groups,with significant difference among them (F =10.210,P =0.012),and the tube number was significantly less in the 500 μg/L neutralizing HspB6 antibody group compared with 0 μg/L neutralizing HspB6 group (P =0.005).The IR of neutralizing HspB6 antibody to the cellular proliferation and migration was enhanced with the increases of concentration and time lapse(Fconcentration =7.485,P =0.002 ; Ftime =16.684,P =0.001).The number of the cells through Transwell chamber membrane was 14.0 ± 2.5,11.1 ± 0.8,6.6 ± 0.1,6.7 ± 0.2 in the 0,50,100,500 μg/L neutralizing HspB6 antibody group respectively,and that in the 100 μg/L and 500 μg/L neutralizing HspB6 antibody group was lessened in comparison with the 0 μg/L neutralizing HspB6 antibody group(both at P=0.000).The apoptosis rate of the cells was (22.73 ± 2.53)％ in the neutralizing HspB6 antibody group,which was significantly lower than (13.33±2.08) ％ of the control group (t=4.967,P=0.008).Conclusions Neutralizing HspB6 antibody inhibits capillary tube formation of human choroidal endothelial cells in vitro in dose-and timedependent manner,probably through suppressing the proliferation and migration and promoting the apoptosis of choroidal endothelial cells.

A pair of degenerate primers were designed based on NBS (nucleotide binding site, NBS) domain of resistance(R) gene and used to perform PCR with cDNA from the translocation line 6VS/6AL of Triticum aestivum-Haynaldia villosa. A clone (N7) characterized with NBS was obtained by sequencing analysis. Two specific primers were designed from the N7 sequence and used to screen a genomic TAC (transformation-competent artificial chromosome, TAC) library of 6VS/6AL consisting of ca. 2 x 10(6) clones. The library was stored as clone pools in twenty-two 96-well plates, each well containing approximately 1000 TAC clones. TAC plasmids were prepared from all the 2112 pools. Using a pooled PCR screening procedure, a positive TAC clone having a 40 kb insert was obtained. The positive clone was confirmed by Southern hybridization with the NBS fragment as a probe. The results indicate that the pooled PCR method is effective for screening of genomic libraries having large number of clones.
Amino Acid Sequence ; Base Sequence ; Binding Sites ; Blotting, Southern ; Chromosomes, Artificial ; Genomic Library ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Translocation, Genetic ; Triticum ; genetics

Congresses as Topic ; Humans ; Rhinitis ; Rhinitis, Allergic, Perennial

OBJECTIVETo investigate the changes and significance of serum gastrin (GAS), beta-endorphin (beta-EP) and plasma motilin (MTL) in patients with severe burns.

METHODSBlood samples were gotten according to different timepoints from 32 admitted burned patients, and then serum GAS, beta-EP and plasma MTL were determined by radio-immuno assay (RIA).

RESULTSIn patients with severe burns, serum GAS decreased significantly in early period. And at the timepoint of 8 h, it reached the lowest level. But during 9-24 h it elevated for a while, and then it reached a relatively stable level. MTL reached the highest level at the timepoint of 2 h after burning. Then at the shock stage, it was comparatively lower. And at the timepoint of 8 h after burning, it reached the lowest level, then raised persistently after reabsorption, but still lower than the normal level. At the early stage after burning, beta-EP raised, then reached the highest level at 8 h after burning. GAS and MTL decreased and beta-EP increased significantly with the increase of the burned area. However, when the burned area was over 70% of the total body surface area, there was no relationship between them.

CONCLUSIONBlood GAS, MTL and beta-EP have represented regular changes in patients with severe burns at the early stage after burning. And the pain-stimulus and shock are effective factors.

Adolescent ; Adult ; Burns ; blood ; Female ; Gastrins ; blood ; Humans ; Male ; Middle Aged ; Motilin ; blood ; Radioimmunoassay ; Time Factors ; beta-Endorphin ; blood

BACKGROUNDSepsis is a leading cause of death in the intensive care units. The late inflammatory cytokine, high-mobility group box 1 (HMGB1), plays a critical role in sepsis. In the present study, we investigated the association between the serum HMGB1 levels and the severity of organ injury in the lipopolysaccharide-induced sepsis in rats.

METHODSTo produce an animal model of sepsis with different degree of organ injury, animals were treated with three different doses of lipopolysaccharide (4, 8 and 16 mg/kg), and the animals in control group were treated with the same volume of the vehicle (saline). The levels of serum HMGB1 were measured at 0, 2, 4, 8, 16, 24, 32 and 48 hours after lipopolysaccharide (LPS) or vehicle injection, meanwhile the biochemical and histopathological indicators for the severity of organ injury were assessed.

RESULTSThe level of HMGB1 had a positive, high correlation with the abnormal changes of serum cardiac troponin I, alanine aminotransferase, aspartate aminotransferase, creatinine and blood urea nitrogen, as well as the pathologic scores of heart, lung, liver and kidney.

CONCLUSIONSThe level of serum HMGB1 is highly correlated with the severity of sepsis in rats, suggesting that HMGB1 could serve as a valuable adjunct in the diagnosis and management of sepsis.

Animals ; HMGB1 Protein ; blood ; Lipopolysaccharides ; therapeutic use ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sepsis ; blood ; drug therapy ; pathology

OBJECTIVETo investigate chloride channel 1 (CLCN1) gene mutation and clinical features of 2 Chinese patients with myotonia congenita.

METHODSClinical data of a patient from a family affected with myotonia congenita in addition with a sporadic patient from Fujian province were analyzed. Exons of CLCN1 gene were amplified and sequenced.

RESULTSThe proband from the affected family was found to carry a c.1024G>A heterozygous missense mutation in exon 8, whilst the sporadic patient has carried a c.1292C>T heterozygous missense mutation in exon 11.

CONCLUSIONDetection of CLCN1 gene mutation is an effective method for the diagnosis of myotonia congenita. Exon 8 of CLCN1 gene may be a mutational hotspot in Chinese patients with myotonia congenita.

Adolescent ; Base Sequence ; Chloride Channels ; genetics ; Exons ; Heterozygote ; Humans ; Male ; Mutation ; Myotonia Congenita ; diagnosis ; genetics ; Pedigree

Objective In order to compare the iodine deficiency disorders(IDD)prevalent slatus in Chun'an County between 2006 and 2007,and to provide the science information for iodine supplementation in different regions.Methods Three schools of Wangzhai,Pingmen and Wenchang which the goiter prevalenee was the most severe were selected in Chun'an County;and from each school,90 pupils aged 8-10 years were randomly selected.B-ultrasound examination of thyroids,urine iodine and salt iodine were measured.Results The goiter rate in B-ultrasound were 7.5%(20/267),median of urine iodine was 247.5 μg/L,mean of salt iodine was 32.7 ms/ks in 2006;and the goiter rate in B-uhrasound were 3.7%(10/271),median of urine iodine was 383.4 μg/L,mean of salt iodine was 33.5 mg/kg in 2007.The goiter prevalence in Wangzhai,Pingrnen and Wenehang township were 15.2%(14/92),6.0%(5/83)and 2.2%(2/92),respectively,and median of urine iodine were 360.1.211.3,189.3μg/L,respectively,in 2006;The goiter prevalence were 6.6%(6/91),3.3%(3/90)and 1.1%(1/90),respectively.and median of urine iodine were 388.6,41 1.5,327.8μg/L,respectively,in 2007.Family ineome of Wangzhai,Pingmen and Wenchang township were 1000,2000,3000 yuan,respectively.Conclusions Goiter prevalence was correlated with urinary iodine,nutritional state and economic condition,high urinary iodine contents and poor nutritional status lcad to a high goiter rate.

AIM To establish the HPLC characteristic chromatogram of Baqi Rougan Decoction reference sample,and to investigate its quantitative transmission laws.METHODS The contents of calycosin 7-O-glucoside,hesperidin,rosmarinic acid,curcumenol and nystose were determined.The transfer rates of decoction piece-aqueous decoction-reference sample were calculated,after which the paste-forming rate and pH value were recorded.RESULTS There were sixteen characteristic peaks in fifteen batches of reference samples with the similarities of 0.90,nine of which were identified.The average transfer rates of nystose and calycosin 7-O-glucoside in the reference sample were(83.14±6.25)%and(77.81±8.31)%,while those of rosmarinic acid and curcumenol in the aqueous decoction-reference sample were(81.71±6.27)%and(72.16±5.91)%,along with the average paste-forming rate and pH value of(38.91%±1.46%)and 5.13±0.08,respectively.CONCLUSION This stable and feasible method can provide a reference for the selection of preparation process and evaluation of key chemical properties for Baqi Rougan Decoction.

Objective: To investigate the relationship between human cytomegalovirus (HCMV) infection and peripheral blood CD14 CD16 monocytes in the pathogenesis of coronary heart disease (CHD), and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection, inflammation, and CHD, to provide a basis for the prevention, evaluation, and treatment of the disease. Methods: In total, 192 patients with CHD were divided into three groups: latent CHD, angina pectoris, and myocardial infarction. HCMV-IgM and -IgG antibodies were assessed using ELISA; CD14 CD16 monocytes were counted using a five-type automated hematology analyzer; mononuclear cells were assessed using fluorescence-activated cell sorting; and an automatic biochemical analyzer was used to measure the levels of triglyceride, cholesterol, high- and low-density lipoprotein cholesterols, lipoprotein, hs-CRp and Hcy. Results: The positive rates of HCMV-IgM and -IgG were significantly higher in the CHD groups than in the control group. HCMV infection affects lipid metabolism to promote immune and inflammatory responses. Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD. The expression of CD14 CD16 mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection. Thus, HCMV antibody as well as peripheral blood CD14 CD16 mononuclear cells can be used to monitor the occurrence and development of CHD.
Angina Pectoris ; epidemiology ; virology ; China ; epidemiology ; Coronary Disease ; epidemiology ; virology ; Cytomegalovirus ; physiology ; Cytomegalovirus Infections ; complications ; Humans ; Incidence ; Inflammation ; epidemiology ; etiology ; Leukocyte Count ; Monocytes ; metabolism ; Myocardial Infarction ; epidemiology ; virology

Objective Based on the finite element method, both sacroiliac fusion and sacroiliac contact models were built to compare the biomechanical differences between the two models and to explore the biomechanical mechanism in the treatment of low back pain by sacroiliac fusion. Methods Two pelvic finite element models were constructed, including the pelvic ring, sacrum, part of the femur, ligaments, cartilage and joint contact. The sacroiliac joints were set to be contact in one model and fusion in the other, respectively. Differences in mechanical conduction on the pelvic ring and the stress on the sacroiliac cartilage under 500 N load between the two models were explored. Results For the fusion model, stresses and displacement on the sacroiliac joint were significantly lower than that of the contact model, especially on the sacroiliac cartilage, where the displacement was reduced by 261% from 0.83 mm to 0.23 mm, and the stresses reduced by 32% from 6.6 MPa to 5.0 MPa. However, the transfer of stress on the pelvic ring was relatively more concentrated in the fusion model. Conclusions Sacroiliac fusion may provide better therapeutic effects on the treatment of low back pain, but the risk of disc herniation and femoral head necrosis must be assessed seriously in advance.