Objective Preparing platelet rich gel through two-times centrifugal technique and co-culturing chondrocytes with PRG, then observing the proliferation and gene expression of chondrocytes, in order to provide a favorable way to prepare tissue engineering cartilage. Methods Centrifugating venous blood of rabbit through two-times centrifugal technique to obtain platelet rich plasma( PRP) ,then detecting the concentration of various growth factor in PRP. Admixing PRP with chondrocytes of rabbit and activating them with activator. After co-culti-vation,the proliferation of chondrocytes through MTT method and expression of ACAN,CollagenⅡand SOX-9 through realtime-PCR were ob-served,and compared with common cultured chondrocytes. Results The concentrations of PDGF-AB,TGF-β1,IGF-1 and VEGF in PRG were significantly higher than those in blood(P<0. 05). After co-cultivation, the proliferation rate of chondrocytes and the expression of ACAN,Collagen Ⅱ and SOX-9 were significantly higher than that of common cultured chondrocytes(P<0. 05). Conclusion Co-culturing chondrocytes with PRG is able to promote the proliferation and gene expression of chondrocytes. We considered that it is a excellent method to construct tissue engineering cartilage.

Objective To observe the roles of nuclear factor-κB (NF-κB) and hypoxia-inducible factor-1 o (HIF-1 α) in hippocampal neurodegeneration of status epilepticus (SE) rats, and explore whether HIF-1α activation is regulated by NF-κB.Methods A total of 110 male Sprague-Dawley rats were randomly divided into seven groups : (1) Control group treated with saline (control, n =15), (2) sham group implanted cannula into lateral ventricle and treated with saline (sham, n =15), (3) SE group treated with pilocarpine (SE, n =20), (4) NF-κB activity inhibitor pyrrolidine dithiocarbamate (PDTC) group treated only with PDTC (PDTC, n =15), (5) SE + PDTC group treated with pilocarpine plus PDTC (SE + PDTC, n =15), (6) SE + HIF-1o siRNA group implanted cannula into lateral ventricle and treated with pilocarpine plus HIF-1 α siRNA (SE + HIF-1α siRNA, n =15), (7) SE + control siRNA group implanted cannula into lateral ventricle and treated pilocarpine plus control siRNA (n =15).SE was induced by injecting lithium chloride and pilocarpine.The seizure of rats was observed.The protein expressions of NF-κB and HIF-1 α in hippocampus of rats were examined by Western blotting.The degenerating neurons in hippocampus were detected by Fluoro-Jade C (FJC) staining.Results Twenty-four hours after termination of SE, the nuclear protein expressions of NF-κB and HIF-1α in hippocampus of rats were increased in SE group (0.57 × 0.06, 0.47 ± 0.07) compared with those in control group (0.23 ± 0.03, 0.20 ± 0.03;P ＜0.05);and compared with SE group PDTC significantly decreased the nuclear protein expressions of NF-κB and HIF-1 α in SE + PDTC group (0.23 ± 0.03, 0.14 ± 0.03;P ＜ 0.05);in SE + PDTC group the numbers of FJC positive cells in CA1 area (28.33 ±5.03) were decreased compared with that in SE group (76.67 ± 13.32);HIF-1 o siRNA injected into lateral ventricle of rats significantly decreased the expression of HIF-1α in hippocampus (0.22 ±0.03) and the number of FJC positive cell in CA1 area (27.34 ±7.02) in SE + HIF-1α siRNA group compared with those in SE group (0.39 ±0.06, 76.67 ± 13.32;P ＜0.05).Conclusions These data suggest that SE can result in activation of NF-κB/HIF-1o pathway in brain.Inhibition of the pathway can attenuate hippocampal neurodegeneration caused by SE, which has the brain protective effect.

[Abstract ] Objective High-mobility group box-1 (HMGB1) is abundantly released in the epileptogenic brain tissue , but few reports are seen about the effect of HMGB 1 on the expression of P-glycoprotein ( P-gp) in the vascular endothelial cells of the epi-leptogenic tissue .This study is to explore whether HMGB 1 can regulate P-gp expression in the brain microvascular endothelial cells of the mouse in vitro . Methods Immortalized brain microvascular endothelial bEnd .3 cells of the mouse were cultured in vitro and al-located to different concentration groups ( treated with culture medium containing 10 , 100 , 500 , and 1000 ng/mL HMGB1 for 8 hours), treatment duration groups (treated with culture medium containing 100 ng/mL HMGB1 for 4, 8, 16, 24, and 32 hours), and a control group ( treated with culture medium without HMGB 1 ) .The mRNA expression of P-gp-encoding gene-multidrug resistance gene 1a (mdr1a) was detected by real-time qPCR, and its protein expression determined by Western blot and immunocytochemistry . Results The results of qPCR manifested that the expressions of mdr 1a mRNA were 1.646 ±0.176, 1.777 ±0.135, 1.617 ±0.043, and 1.398 ±0.182 in the 10, 100, 500, and 1000 ng/mL HMGB1 groups, respectively, significantly higher than 1.030 ±0.284 in the control group (P<0.05), and so were those in the 4, 8, 16, 24 h, and 32 h groups (2.655 ±0.112, 2.168 ±0.212, 1.823 ± 0.232, 1.418 ±0.376, and 1.445 ±0.123) than in the control (1.010 ±0.164) (P <0.05).Western blot showed a significant increase in the P-gp protein expression in all the concentration groups (P<0.05) as well as in the 8 h and 16 h treatment duration groups as compared with the control group (P<0.05).Immunocytochemis-try also revealed a higher P-gp expression in the HMGB1-treated than in the control cells (P<0.01). Conclusion HMGB1 can upregu-late the expressions of mdr1a mRNA and P-gp protein in the brain microvascular endothelial cells of the mouse , which may associated with drug resistance of central nervous system diseases , especially that of epilepsy .

BackgroundThe study of myopia development is always the hotspot worldwide. Recently,scientist found that some growth factor secreted by retinal nerve epithelium cells and retinal pigment epithelium(RPE)cells are associated with the development of myopia. Whenever, the absorption of RPE cells to different wave-length lights is different. ObjectiveThis study was to investigate the effects of different wave-lengths lights on the proliferation of human RPE cells, and explore the influence of different wave-lengths lights on RPE cells secreting hepatocyte growth factor(HGF) ,basic fibroblast growth factor(bFGF) and transforming growth factor-β(TGF-β).Methods The fourth to fifth passages of human embryonic RPE cells were exposed to blue light( λ =480 nm),red light( λ =775 nm) and white light. The cells of control group were harvested in normal condition. The proliferation and growth of RPE cells were assayed by MTT,and the ultrastructure of cells was examined under the transmission electron microscopy at 48 hours after light exposure of RPE cells. Enzyme linked immunosorbent assay(ELISA) was adopted to determine the concentrations of HGF,bFGF and TGF-β in the culture medium in 12,24,48,72 hours. The expression of HGF mRNA in RPE cells was detected by RT-PCR. This study was approved by Ethic committee of Fudan University. ResultsThe A490 values of the cells exposed to blue light,red light,white light and white light were 0. 0218±0. 0014 ;0. 0353±0. 0025 ;0. 0371 ±0. 0024 and 0. 0445 +0. 0046 respectively with the significant difference among 4 groups ( F =12. 579, P＜0.05 ), and A490 value in blue light group, red light group were significantly lower than that of the control group ( t =2.043 ; t =2.024, P＜0.05 ). ELISA showed that the concentrations of HGF and TGF-β in culture medium were evidently elevated as the prolongation of light exposure in various light exposure groups in 72 hours(HGF) and 48 hours(TGF-β) compared with 12 hours with a predominating rise in the control group. The statistically significant differences were found in the concentrations of HGF and TGF-β between control group and blue light group or red light group in the( all P＜0. 05 ). The bFGF level was decreased with the time increase of various light exposure with the significant differences in 72 hours compared with 12 hours( P＜0.05 ). RT-PCR revealed the considerable difference about expression of HGF mRNA in RPE cells among these four groups( P＜0. 05 ), and the lest expression in HGF mRNA was in the blue light group compared with control group( t =3. 972,P＜0.05 ). Thinning of the chromatin, decreasing of organelle and loss of cellular membrane were seen in the blue light group, but no obvious change of ultrastructure of human embryo RPE cells was found in the ret and white light groups. ConclusionsThe irradiation of different wave-length light can effect the growth and proliferation and secretion of HGF,bFGF and TGFβ in human RPE cells in vitro,implying myopia formation is associated to exposure of different wave-length light.

Acute Disease ; Adult ; Aged ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Young Adult

Objective To analyze the expression of the placenta Grb10 from women conceived by transferred thawed blastocyst,and to evaluate the security of blastocysts vitrification.Methods A cross-sectional study was performed in the Department of Obstetrics and Gynecology of Fujian Provincial Maternity and Children's Hospital from January 2012 to May 2014,50 women conceived by transferring thawing blastocyst and 50 natural pregnancy control women were enrolled in this study.The expression of Grb10 protein was detected by immunohistochemistry and Western blot,and the expression of Grb10 mRNA was detected by Realtime PCR method.Results Comparison of two cases of gestational age,gestational age,fetal sex,fetal body weight,body length,head circumference,abdominal circumference,there were no significant differences(P＞0.05),comparison of placental area,placental weight,the difference was statistically significant(P＜0.05).Real-time PCR and Western blot results showed that,there was no significant difference in the expression of Grb10 mRNA and protein between the two groups(P＞0.05).Conclusion Blastocysts vitrification may increase the area and quality of delivery of placenta,however,there was no significant change in the expression of Grb10 in placenta.

AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTM). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTM were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-?) and interleukin-6(IL-6) in the hTM was analyzed by radioimmunoassay, after hTM were cultured with 0.312,0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTM induced by PPM, and the levels of TNF-? and IL-6 in the hTM induced by PPM increased markedly too. And the induced expression effect of TNF-? and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTM,and activate hTM in vitro.

The genotypes of extended spectrum β-laetamases(ESBLs) and AmpC β-lactamases produced by Enterococcus gallinarum isolated from chloebia gouldiae were determined to elucidate the evolution mechanism of the resistant genes.The minimum inhibitory concentrations (MICs) of 18 antibacterial drugs against the Enterococcus gallinarum were detected with two dilution method,and the ESBLs and AmpC β-lactamases from the bacterium were amplified by PCR using the primers of TEM,SHV,CTX-M,ACC,CIT,DHA,EBC,FOX and MOX,respectively.The PCR products were cloned and then the cloned fragments were sequenced to identify their genotypes and subtypes.The bacterium was proved to be a ESBL-producing and AmpC β-lactamase-producing bacterium,showing severe resistant to the other drugs,except the third and forth cephalosporins,carbopenems and fosfomycin.Compared with that of AJ847364 (TEM-116),the sequence of the TEM-type was characterized by two nucleotide mutations (512T→A and 695A→C),which led to two mutations of amino acids(17111e→Lys and 232Lys→Thr),showing that the detected TEM-type was a new genotype,the sequence of the AmpC β-lactamase was similar to that of EF078894 (ACT-like type)with a 97% homology.The genotype of ESBLs of Enterococcus gallinarum was a new TEM-type derived from the TEM-type ESBLs of klebstella pneumoniae isolated from the same avian.The genotype of AmpC lactamase was ACT-type,which probably concerned with β-1actam antibiotics used.

Objective To determine the normal range of minimal erythema dose (MED) for ultraviolet A (UVA) and B (UVB) in volunteers from Urumqi region.Methods One hundred and twenty-seven volunteers including healthy subjects and patients with noninflammatory skin disorders were enrolled in this study.SUV-1000 type UV simulator was used as the light source to determine MED of UVA and UVB in these subjects.Results These subjects included 48 persons with Fitzpatrick skin type Ⅲ,79 with Fitzpatrick skin type Ⅳ,51 males and 76 females.The median MED value for UVA and UVB was 38.1 J/cm2 and 31.8 mJ/cm2 respectively in subjects with skin type Ⅲ,59.16 J/cm2 and 48.00 mJ/cm2 respectively in subjects with skin type Ⅳ.Significantly lower median MED values of UVA (both P ＜ 0.01) and UVB (both P ＜ 0.05) were observed in the male and female subjects with skin type Ⅲ compared with those with skin type Ⅳ.The male subjects showed a significantly higher median UVA-MED value (59.16 J/cm2 vs.41.10 J/cm2,P ＜ 0.05),but a similar UVB-MED value (39.60 mJ/cm2 vs.35.55 mJ/cm2,P ＞ 0.05) compared with the female subjects.No significant difference was observed in the median value of UVA-or UVB-MED in subjects with skin type Ⅲ or Ⅳ between Han and Uygur nationality (all P ＞ 0.05).Also,no correlation was found in the median value of UVA-or UVB-MED with age or duration of outdoor exposure in the male or female subjects (all P ＞ 0.05).The lower reference limit was 33.38 J/cm2 for UVA-MED and 27.90 mJ/cm2 for UVB-MED in the population in Urumqi region.Conclusion Skin phototype may be an important determinant of MED.

Objective To study the the feasibility of real-time 3D transesophageal echocardiography (RT-3D TEE) in measuring the distance between the coronary artery ostia and the aortic annulus.Methods RT-3D TEE image of aortic root was collected intraoperatively in 18 cases who were refered to our center for transcatheter aortic valve implantation.The aortic annulus was reconstructed and the height of the annulus (H) was generated.Meanwhile RT-3D TEE and MDCT was used to determine the left coronary artery ostia-aortic annulus distance (E-LAA,C-LAA) and the right coronary artery ostia-aortic annulus distance (E-RAA,C-RAA).Results The display rate of right coronary artery was 94.4％ (17/18),with the left coronary artery 88.9％ (16/18) on the RT-3D TEE imaging.The height of the annulus(H) was lower than the distance between the coronary artery ostia and the aortic annulus(P ＜0.05).No statistically significant differences was found in the parameters of the distance between the coronary artery ostia and the aortic annulus between RT-3D TEE and MDCT (P ＞0.05).Conclusions RT-3D TEE imaging can display and measure the distance between the coronary artery ostia and the aortic annulus.