Angiogenesis, or the formation of new blood vessels out of pre-existing capillaries, is very important in many physiologic and pathologic processes, such as embryonic development, cancer, retinopathies, etc. Vascular endothelial growth factor receptor-2 (VEGFR-2) plays a key role in angiogenesis. In this review, we discussed the structure, function and signal transduction of vascular endothelial growth factor receptor-2. Understanding these should provide important insights into how new strategies can be devised to interfere in the physiologic and pathologic processes involved in angiogensis. [

Mitochondrion is known as the energy factory of the cell, which is also a unique mammalian organelle and considered to be evolved from aerobic prokaryotes more than a billion years ago. Mitochondrial DNA, similar to that of its bacterial ancestor’s, consists of a circular loop and contains significant number of unmethylated DNA as CpG islands. The innate immune system plays an important role in the mammalian immune response. Recent research has demonstrated that mitochondrial DNA (mtDNA) activates several innate immune pathways involving TLR9, NLRP3 and STING signaling, which contributes to the signaling platforms and results in effector responses. In addition to facilitating antibacterial immunity and regulating antiviral signaling, mounting evidence suggests that mtDNA contributes to inflammatory diseases following cellular damage and stress. Therefore, in addition to its well-appreciated roles in cellular metabolism and energy production,mtDNA appears to function as a key member in the innate immune system. Here, we highlight the emerging roles of mtDNA in innate immunity.
Animals ; DNA, Mitochondrial ; genetics ; Humans ; Immunity, Innate ; immunology ; Mitochondria ; genetics ; immunology ; Signal Transduction

Objective To understand the iodine nutrition of children in Qiandongnan and to provide the scientific evidence for prevention and control of iodine deficiency disorders (IDD). Methods The 3200 urinary samples of pupils aged 8-10 were collected from 80 primary schools in 16 county and city in Qiandongnan in 2006. The urinary iodine content was tested by colorimetric ceric-arsenic assay and vitriolic ammonium assimilation. Results The median of urinary iodine content in children samples was 362.9?g/L, the percentage of urinary iodine( ≥100?g/L) was 98.00%, in which ≥300?g/L was 65.31%, the percentage of urinary iodine(

Objective Based on the teeth in vitro skull specimen to investigate the feasibility and accuracy of CARE kV CT scan for reducing radiation dose. Methods Of eight fixed and preserved skull teeth were scanned repeatedly in groups of A, B, C. In expose factors of 100 kVp/150 mAs in conventional scanning of group A, open CARE Dose 4D scan of group B and CARE kV scan of group C. We assessed the radiation dose in the group A, B and C, the average CT values of the images, noise, SNR, CNR and subjective ratings. Results Three groups of actual scan tube current, tube voltage and radiation dose were statistically significant (P < 0.05). Compared with group A, the radiation dose in the group B was reduced by about 13.76% and 26.61%reduced in the C group. The average CT values of the images and the noise SNR, CNR and subjective ratings hasd no significant difference (P > 0.05). Conclusion CARE kV technology based on the use specimens tooth can reduce the radiation dose, protect image quality, and be consistent to the reports of previous researches.

Objective To investigate the performance of dual source CT scanning combined with low-tube-voltage iterative reconstruction (SAFIRE) for demonstrating coronary artery calcium plaques. Methods 70 patients with clinically suspected coronary atherosclerosis and with normal BMI received SAFIRE-3 or filtered back projection (FBP) for reconstruction after undergoing CT scans with tube voltage range of 100 kV. The CT images (average CT values, noise, SNR, and CNR) and subjective scores (number of calcium plaque, calcium plaque adjacent artifacts, and image quality) were compared between the two groups. Results The average CT value of the aortic root did not differ significantly between the two procedures for reconstruction (P > 0.05). Ob jective evaluation of the images (noise, SNR, and CNR) between the two groups was statistically significant (P < 0.05 for all comparisons), so was image subjective rating (number of calcium plaque and rate of calcium plaque edge)(P < 0.05 for all comparisons). SAFIRE-3 reconstruction was markedly superior to FBP reconstruc-tion. Conclusions Low kilovoltage combined with reconstruction of SAFIRE-3 can reduce calcium plaque adja-cent artifacts and enhance quality of images , which is better than FBP in displaying the morphology and adjacent conditions of calcified plaque; therefore it is worth popularizing.

Objective To study the role of epidermal growth factor (EGF),epidermal growth factor receptor(EGFR),extracellular signal-regulated kinase 1/2 (p-ERK1/2) in the pathogenesis of endometriosis under estrogen deprivation conditions.Methods The estrogen was quickly-stripped in medium and the female nude mice were castrated by bilateral oophorectomy to build estrogen deprivation in vitro and in vivo experimental models,respectively.(1) In vitro experiments:according to different treatments the estrogen deprived ectopic endometrial cells were classified into 4 groups:①EGF group:the ectopic endometrial cells were cultured for 72 hours with different concentrations of EGF (0.01,0.1,1,10,50,100 ng/ml),the results of EGF group were represented by the result of cells treated by 10 ng/ml EGF cultured for 72 hours ; ②EGF + PD98059 group:the ectopic endometrial cells were cultured for 72 hours with 5 × 10-2 mol/L PD98059 (inhibitor of ERK),followed by a cultivation for 72 hours treated by 10 ng/ml EGF + 5 × 10-2 mol/L PD98059 ; ③EGF + ICI182780 group:the ectopic endometrial cells were cultured for 72 hours with 10-6 mol/L ICI182780 [inhibitor of estrogen receptor(ER)],followed by a cultivation for 72 hours treated by 10 ng/ml EGF + 10-6 mol/L ICI182780; ④Blank control group:the ectopic endometrial cells were cultured with no treatment.The proliferation activity of ectopic endometrial cells in all groups after treatment were examined by methyl thiazolyl tetrazolium (MTT) method represented by absorbance value (A).The expression of p-ERK1/2 protein were detected by western blot.(2) In vivo experiments:64 female nude mice were randomly divided into control and castration groups (both n =32) using random number chart.The mice in castration group were castrated by bilateral oophorectomy on 3 weeks after the endometriosis model was established.The levels of EGF,EGFR,p-ERK1/2 protein in ectopic lesions of both groups were measured on 4,6,8 and 10 weeks after the endometriosis model was established by western blot.Results (1) The proliferation activity of ectopic endometrial cells:the proliferation activity of ectopic endometrial cells treated by different concentrations of EGF (0.01,0.1,1,10,50,100 ng/ml) for 72 hours were 0.310 ± 0.010,0.340 ± 0.020,0.670 ± 0.010,0.980 ± 0.030,1.360 ± 0.020,1.670 ± 0.020,respectively,the proliferation activity was increased along with of EGF concentrations.The proliferation activity was 0.680 ± 0.030 at EGF + PD98059 group,the differences exhibited significant difference when compared with that at EGF group with 100 ng/ml for 72 hours(P ＜0.01).The proliferation activity of EGF + ICI182780 and blank control groups were 0.330 ±0.030 and 0.310 ±0.030,respectively,which did not reached statistical differences(P ＞ 0.05).(2) The expression of EGF,EGFR,pERK1/2 protein:① In vitro experiments:the levels of p-ERK1/2 protein in EGF and blank control groups were 0.670 ± 0.020 and 0.600 + 0.010,respectively,which reached statistical differences (P ＜ 0.05).The level of p-ERK1/2 protein in EGF + PD98059 group was 0.610 ± 0.020,which exhibited significant differences with that at blank control group(P ＞ 0.05).② In vivo experiments:at 4,6 and 8 weeks after the endometriosis models were established,the expression of EGF protein in the ectopic lesions of castration group and control group were (0.530±0.015 versus 0.610 ±0.015),(0.400 ±0.029 versus 0.620 ±0.018),(0.560 ±0.026versus 0.630 ± 0.021),respectively,the levels of EGFR protein were (0.500 ± 0.030 versus 0.640 ±0.030),(0.470 ± 0.020 versus 0.630 ± 0.020),(0.510 ± 0.030 versus 0.610 ± 0.020) respectively,and the level of p-ERK1/2 protein were (0.500 ± 0.020 versus 0.580 ± 0.020),(0.490 ± 0.020 versus 0.580 ±0.020),(0.570 ±0.020 versus 0.590 0.020),respectively.The difference of EGF,EGFR,pERK1/2 protein expression levels between two groups did not exhibited significant difference(P ＜ 0.01,P ＜0.01,P ＜0.05).At 10 weeks after the endometriosis models were established,the levels of EGF protein in castration group and control group were both 0.620 ± 0.020,the levels of EGFR protein were both 0.610 ±0.020,and the level of p-ERK1/2 protein were 0.590 ±0.010 and 0.600 ± 0.020.No statistical difference (P ＞0.05) was found between those two groups (P ＞ O.05).Conclusions EGF could stimulate the proliferation of ectopic endometrial cells by activating the ERK pathway under estrogen deprivation conditions.The inhibition of EGF signaling system in ectopic lesions was alleviated along with the prolongation of the period of estrogen deprivation.

ObjectiveTo investigate the method of establishing damaged endometrial stromal cells (ESC) model in vitro.Methods ( 1 ) From June to December 2011 ESC from normal endometrim at proliferation phase ( n =8 ) and secretory phase ( n =8 ) were isolated,cultured and identified in vitro.( 2 ) ESC was treated with different concentrations of mifepristone or withdrawal of mifepristone at different time point.The proliferation inhibition percent was measured by cell counting kit-8 ( CCK-8 ). ( 3 ) 0 μmol/L (control group)and 60 μmol/L(experimental group) concentration of mifepristone was added into ESC for 48 hours,then withdrew of mifepristone,continued to be cultured for 48 hours.The morphological changes were observed and apoptosis of ESC in different menstrual cycle were detected by flow cytometry.The mRNA and protein level of vascular endothelial growth factor ( VEGF),caspase-3,8,and 9 were determined by one-step quantitative real-time PCR (Q-PCR) and western blot.Results( 1 ) ESC from 16 specimens of endometrium were all isolated and cultured successfully. (2) The proliferation inhibition rate of ESC was correlated with concentration and duration of mifepristone positively. The proliferation of ESC could be recovered at a range of time after withdrawal of mifepristone.However,when the concentration of mifepristone was 100 μmol/L,the growth of ESC recovered very hardly. (3) The damaged ESC spacing increased,the spindle shape and vacuolization in the cytoplasm were observed in experimental group; the rate of apoptosis of these damaged cells was significantly increased compared with control groups,which were (52 ± 12)％ vs.( 13 ± 5 ) ％ at the proliferative phase and (53 ± 6) ％ vs.( 32 ± 3 ) ％ at the secretory phase ( all P ＜0.05).The relative mRNA level of VEGF was 0.52± 0.12 in experimental group and 1.00 ± 0.17 in control group at proliferation phase (P ＜0.05).And the relative mRNA level of VEGF was 0.19 ±0.03 in experimental group and 0.81 ±0.07 in control group at secretory phase (P ＜ 0.05).The relative level of VEGF protein in the experimental group were both decreased 1.98 and 2.79 folds at the proliferation phase and the secretory phase when compared with those in control group,respectively ( P ＜ 0.05 ).While the relative levels of caspase-3,8,9 mRNA were 5.62 ± 0.65,5.41 ± 0.53,7.22 ± 0.51 in the experimental group and 1.00 ± 0.44,1.00 ± 0.21,1.00 ± 0.32 in control group at the proliferative phase.In the mean time,the relative levels of caspase-3,8,9 mRNA were 10.22 ± 0.72,25.3 ± 1.72,9.48 ± 1.89 in experimental group and 1.42 ± 0.14,1.14 ± 0.28,1.16 ± 0.12 in control group at the secretory phase,respectively (P ＜ 0.05).Compared with the control group,the levels of caspase protein in the experimental group were increased 2.04 and 1.60 folds in caspase-3,4.23 and 1.49 folds in caspase-8,2.65 and 3.5 folds in caspase-9 at the proliferative phase and at the secretory phase,respectively (P ＜ 0.05 ).ConclusionThe damaged model of ESC can be established after 48 hours by the withdrawal of 60 μmol/L mifepristone in treatment of ESC for 48 hours.

Objective To reduce the animal component contamination for human embryonic stem cells ( hESCs ) and to simplify hESCs culture process , we develop a new coating substrate which can support the hESCs growth without dif -ferentiation, and is easy to store and use. Methods Mouse embryonic fibroblasts(MEF)were fixed on the surface of plate by methanol.hESCs were cultured on this new substrate and were passaged every 5 to 6 days.After 10 passages, we checked the cell morphology , alkaline phosphatase expression , embryonic specific markers and the differentiation ability in vitro.Results After 10 passages , the hESCs grew well on this new substrate and maintained the typical hESCs morpholo -gy.Alkaline phosphatase staining was positive .Immunofluorescence staining showed that the expressions of Oct 4, SSEA4, Tra-1-60 were positive .The cells formed embryoid body in vitro .Conclusions This methanol-fixed MEF substrate can support the growth of undifferentiated hESCs .The coating material can be produced in large scale and stored for a long time.It provides a new and relatively easy way to amplify hESCs .

The immune system of mammalian has developed sophisticated mechanism to deal with diverse unknown antigens by random rearrangement of V, D and J antigen gene fragments. The immune self-tolerance is the mechanism to avoid self-destruction by the gene rearrangement generated autoreactive lymphocytes. Until recently it was believed that autoreactive lymphocytes are either deleted or rendered unable to respond by the supposed cell or clone selection mechanism. However, recent findings from a number of laboratories suggest instead of cell selection but receptor selection plays an essential role in immune self-tolerance. Receptor selection is carried out by secondary or nested rearrangement of antigen receptor gene fragments, and can occur at different stages of lymphocyte differentiation. Furthermore, secondary rearrangement of receptor gene also plays an important role in shaping immune response after the interaction of receptor with antigen by altering its specificity.
Autoimmune Diseases ; immunology ; B-Lymphocytes ; immunology ; Gene Rearrangement, B-Lymphocyte ; immunology ; Genes, Immunoglobulin ; Humans ; Immune Tolerance ; Receptors, Antigen, B-Cell ; immunology ; Self Tolerance ; immunology

The purpose of this study was to clone human HSP90beta cDNA and construct its eukaryote expression vector . The total RNA was isolated by TRIzol Reagent (Invitrogen) from human NPC and its cDNA was gained by RT-PCR. The purified RT-PCR products and PGEM-T Easy Vector were ligated and transformed into XL1-blue E. coli bacteria. The white clones were selected and the plasmid was purified, which was further identified by double enzyme digestion and sequenced. PGEM-hHSP90beta and pcDNA3.1(+) DNA were digested by AflII and Xbal respectively. After purification, the two fragments obtained were ligased by using T4 DNA ligase (Fermentas). This recombinant DNA was then transformed into E. coli Competent Cells XL1-blue and positive clones were selected on the LB agarose plate containing Ampr (100 microg/ml). Single clones were identified by double digestion with AflII and Xbal, and two fragments with the size 5.4 kb and 2.1 kb were produced as expected. The hHSP90beta gene was successfully inserted into the eukaryote expression vector pcDNA3.1(+) by the recombination technique in vitro.
Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics