Objective To analyze the human cellular repressor of E1A-stimulated genes (hCREG)using bioinformatics tools and predict the promoter position and transciption factor binding sites.Methods Complete coding hCREG sequences were obtained from Genbank.hCREG was analyzed with First EF software in order to obtain the promoter sequence of hCREG.Moreover,transcription factor binding sites of hCREG was convinced by Motif software.Subsequently,the transcription factor of hCREG by bioinformatics tools was related between hCREG and smooth muscle cells differentiation observed by both Western blot and immunoflourescence.Results The promoter of hCREG was located in -109~-359 bp of up-stream of transcriptional site,which was predicted with 945bp length.Transciption factor binding sites of hCREG was predicted by Motif software which was found with 80 transciption factors.The expression of hCREG and smooth muscle ?-actin(SM ?-actin)increased in HITASY after 72 hours with serum deprivation detected by both Western blot and immunoflourescence.Meanwhile,the results showed that the expression of wtp53 increased significantly.These results suggested that transcription factor wtp53 might regulate the expression of hCREG and promoted dedifferentiated phenotype of VSMCs.Conclusion The transcriptional information of the proximal promoter of hCREG obtained.As up-stream regulational factor of hCREG,wtp53 can up-regulate the expression of hCREG and may play a vital role in the process of VSMCs differentiation and phenotype modulation.

A satisfaction survey carried out by Beijing Chao-Yang Hospital during the Beijing 2008 Olympic Games, found differences of patient satisfaction between locals and overseas patients in consulting environment, outpatient procedure, quality of service, quality of diagnosis and treatment. The patient satisfaction of overseas patients was lower than locals. Especially noteworthy is the low satisfaction of the former for the hospital coloring, privacy in the sector, registration, vocal communication, patient condition statement, nurse attitude and service quality. Real-time improvement in these aspects of medical services has harvested obvious progress. Results of the satisfaction survey provided valuable reference materials for large-scale activities, and ongoing improvement of medical services in the long run.

Sleep plays an important role in the modulation of endocrine function and energy metabolism;hormone levels are markedly different during a sleepless night compared with a night of normal sleep.Sleep deprivation may change the levels of cortisol,growth hormone(GH)and thyrotropin(TSH)at night.In addition,sleep curtailment disturbs the balance of anorexi- genic(leptin)and orexigenic(ghrelin)factors,and does harmful to glucose tolerance.

Objective To explore the effects of early growth response gene-1 (Egr-1) on bone marrow mesenchymal stem cells (BMSC) proliferation and osteogenic differentiation,which is aimed at providing new molecular targets for the treatment of osteoporosis.Methods Bone marrow was collected from adult men and the BMSCs were cultured primarily and observed by microscope.Meanwhile,flow cytometry was used for BMSCs phenotypic identification;After transfection of pcDNA3.1/Egr-1 into BM SCs,the level of BMSCs proliferation was determined by MTT respectively on the 2 d,4 d and 6 d;On the 7 d after transfection,the ALP activity assay was used for testing the ALP activity in BMSCs.And then,alizarin red S-calcium kit was used for measuring the calcified knots respectively on the 7 d,14 d and 21 d;On the 21 d after transfection,real-time qPCR and Western blotting were used respectively for measuring the expression of mRNA and protein of Egr-1,Runx2 and NDRG1;Further,BMSCs were transfected with Egr-1 siRNA,and the content of calcium nodules,ALP activity,the expression of Egr-1,Runx2 and NDRG1 were detected as above methods.Results The cells cultured in vitro showed high level of CD90 and CD29 and very low level of CD34 and CD45,which is accorded with the characteristic of BMSCs.The pcDNA3.1/Egr-1 transfection for BMSCs had no effect on cells prolifera tion.However,the calcified knots,ALP activity and the expression of Egr 1,Runx2 and NDRG1 were increased after transfection of pcDNA3.1/Egr-1 for BMSCs.In addition,Egr-1 siRNA showed the opposite effect with pcDNA3.1/Egr-1 transfection for BMSCs.Conclusion Egr-1 induces osteogenic differentiation of BMSCs by promoting NDRG1 but has no effects on proliferation of BMSCs.

Objective:To observe the changes of visual evoked potential(VEP)in diabetic rats and the influence of nerve growth factor(NGF)and aminoguanidine(AG)on VEP.Methods:Diabetes was induced in adult male Wistar rats with streptozotocin(STZ).Rats were divided into normal control group(CON),diabetes model group(DM).NGF-treated group(D +N)and AG-treated group(D+A).VEP was measured during the 3~(rd)month,6~(th)month,9~(th)month,and 12~(th)month.Results: Compared with the CON group,all rest groups had longer latencies and lower amplitudes(P

The effects of pretreatment methods of Jerusalem artichoke tubers on microbial lipids fermentation with an oleaginous yeast strain Rhodosporidium toruloides Y4 were investigated in shaking flask culture.The yeast strain accumulated substantial amount of lipids using either purple-or white-skinned Jerusalem artichoke tubers as sole carbon and energy source.When cells were cultured on the extracted juice or the acidhydrolysate,cellular lipid content reached 40%(w/w),while cultured on the pulp,the white-skinned tubershadhigher lipid productivity,yielding 12.1 g lipids per100 g dried tubers.Major fatty acid constituents of microbial lipids were those contained 16-and 18-carbon atoms based on GC analysis,which is quite similar to traditional vegetable oil.Microbial lipids prepared from Jerusalem artichoke can be applied to biodiesel production.

Objective To investigate the clinical features and genetic basis of cryopyrin-associated periodic syndrome (CAPS). Methods The clinical manifestations, laboratory tests, and genetic tests of one case of CAPS were retrospectively analyzed. The related literatures were reviewed. Results A 7 year and eight month old male patient had recurrent fever for 7 years and his whole body was covered with patchy red rash which was itchy and faded with pressure. The limbs and joints were normal. The levels of high-sensitivity C-reactive protein, erythrocyte sedimentation rate, rheumatoid factors were increased. The patient had fundus arteriosclerosis, double conjunctival lesions and nerve deafness on both sides. There was no mutation found in NLRP3 gene coding region, but a heterozygous mutation (-2667G>T) had been found in 5 ' untranslated region. Compared with normal control, the mRNA level of NLRP3 increased 4.2 times and the expressions of IL-1βand IL-18 gene increased 2.2 (P=0.002) and 1.2 times (P>0.05). Conclusions The clinical features of CAPS can be recurrent fever, rash, and joint involved. The oph-thalmologic abnormalities and varying degrees of deafness may occur during the progression. The test of NLRP3 gene may help diagnosis.

BACKGROUND: Research on seed cells is the most important aspect in the filed of tissue-engineering research, and because of their various ad vantages, bone marrow mesenchymal stem cells (BMSCs) have been taken as the ideal bone tissue-engineering seed cells. OBJECTIVE: To observes the growth characteristics of in vitro cultured BMSCs and their osteogenetic characteristics under non-inducing conditions. DESIGN: A single sample experiment.SETTING: Implanting Center of Stomatological Hospital of Fujian Medical University MATERIALS: This experiment was conducted at the Central Laboratory of Stomatological Hospital of Fujian Medical University, between May andDecember 2003. BMSCs are derived from the primary passage to the 3rd passage of in vitro cultured cells from 4 one-year old dogs. METHODS: Under the aseptic conditions, bone marrow puncture was made at bilateral femur trochanter and 5 mL of heparin anticoagulatedbone marrow was obtained. Then it was placed in 50 mL of anti-Dulbecco's modified Eagle's culture medium in centrifuge tube for monocyte isolation. The BMSCs single cells were primarily isolated and placed in culture box for subculture. After 48 hours, culture medium was removed and the medium was cha1ged once every 3 days. Then subculture was carried on continuously to observe BMSCs in morphology under inverted the phase contrast microscope with the assistance of HE staining. The number of cells was counted daily to calculate doubling time and to draw a growth curve. Alkaline phosphatase was detected by using calcium-cobalt method,and chinalizarin staining was used to detect the growth state of calcification tubercle. MAIN UTCOME MEASURES: ①Optical microscopic structure of dog BMSCs; ② The growth curve of dog BMSCs; ③Observation of osteogenetic index- alkaline phosphatase and calcification tubercle. RESULTS: ① Morphological observation indicated that BMSCs were adherent to the walls, clonogenic and appearing fibroblastic phenotype, and they presented morphological changes without exposing to osteogenetic inducer. ② The expression of Alkaline phosphatase in primary cells was stronger, and it was strong in the 1st passage cell, but weak in the 2nd and 3rd passage cells. ③Calcium deposition was observed, which was stronger in primary cells than in subcultured cells. CONCLUSION: ①BMSCs massively proliferated during in vitro culture,capable of differentiating into osteoblasts and considered as the optimal seed cells for bone tissue-engineering reconstruction. ② BMSCs derived from the 3rd passage has osteogenic activity, but the osteogenic activity of the primary cultured cells was stronger than thatr of the subcultured cells.

BACKGROUND: Three-dimensional printing technology is a new technology which can quickly and accurately transform the virtual computer-aided design into the three-dimensional physical prototypes. Three-dimensional printing physical model method can replace the method of traditional preoperative planning and repair surgical simulation, with the characteristic of repeatable, which has been deepened day after day in clinical application of spine surgery. OBJECTIVE: To summarize the application status of three-dimensional printing technology in spine surgery and look forward to its future development directions. METHODS: The articles regarding the application of three-dimensional printing technology on clinical applications in spine surgery were retrieved from PubMed databases, Google Scholar, China National Knowledge Infrastructure and Wanfang Database from January 2000 to July 2015. The key words were 3D printing technology, rapid prototyping technology, spine, vertebra, department of orthopedics, fracture, joint, hand and foot, bone tumor, trauma, cervical vertebrae, thoracic vertebrae, lumbar vertebrae, sacral vertebrae, pedicle of vertebral arch, vertebral body, intervertebral disc, and clinical application. A total of 50 articles with a good representation were selected and discussed after repetitive studies and reviews were excluded. RESULTS AND CONCLUSION: The three-dimensional printing technique has been applied in preoperative diagnosis, individualized orthosis customerization, the communication between doctors and patients, teaching, the formulation of individualized and high-accurate repairing plan, intraoperative navigation and individualized implant customization. These results suggest that with the rapid development of medical imaging, digital medicine and technologies of the cel and tissue culture and new materials, three-dimensional printing technology wil have a wide range of applications in spine surgery.

Objective To determine the effect of Leptospira interrogans lipopolysaccharide (L-LPS) inducing apoptosis of murine mononuclear-macrophage cell line( J774A. 1 ), and apoptotic regulation of Toll-like receptor(TLR) and associated intracellular signaling pathways. Methods Lipopolysaccharide (L-LPS) of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai 56601 was prepared using phenol-water method. The effects of L-LPS inducing J774A. 1 cell apoptosis and the apoptosis-blocking with FasL neutralizing antibody were detected by flow cytometry. Real-time fluorescent quantitative RT-PCR (qPCR) and flow cytometry were performed to measure the changes of Fas/FasL mRNA and protein expression levels in J774A. 1 cells before and after L-LPS treatment. The regulations in L-LPS-induced cell apoptosis by TLR2 and TLR4 as well as p38MAPK, JNK, ERK pathways were determined by either TLR2 or TLR4 antibody blocking test, signaling pathway blocking test and flow cytometry. Results 56.50%, 69.28% and 24.35% of the J774A. 1 cells after treatment with 100 ng/ml L-LPS for4, 12 and 24 h were apoptotic,while the apoptosis rates were decreased to 11.21%, 21.58% and 12.70% after the cells blocked by FasL neutralizing antibody(P ＜0.05). The levels of FasL and Fas mRNAs in J774A. 1 cells treated with L-LPS for 4, 12 and 24 h were elevated with 1.34, 2.12, 2.10 times and 2.45, 3.87, 3.12 times compared to those in the L-LPS untreated cells (P ＜ 0. 05 ), respectively, while the expression rates of FasL and Fas proteins were upregulated to 18.61%, 60.13%, 42.75% and 76.34%, 85.70%, 77.92% from 4.82% and 15.32% apoptotic rates in the L-LPS untreated cells, respectively( P ＜0.05 ). The L-LPS-induced apoptosis rate( 11.54% ) of TLR2 antibody blocked J774A. 1 cells was significantly lower than that(66.56% ) of the J774A. 1 cells without TLR2 antibody blocking( P ＜0.05 ), but L-LPS-induced apoptosis rate of TLR4 antibody blocked J774A. 1 cells was as high as 55.27% ( P ＞ 0.05 ). Compared to the apoptosis rate (62.17%) in the p38MAPK and JNK pathway-free J774A. 1 cells, the L-LPS-induced apoptosis rates in p38MAPK blocked cells(20.54% ) and JNK blocked cells(47.98% ) were significantly lower( P ＜0.05 ),and the apoptosis rate in ERK blocked cells was as high as 61.72% ( P ＞ 0.05 ). Conclusion L-LPS was recognized by TLR2 and upregulates both Fas and FasL expression via p38MAPK and JNK pathways, which involving in the process of the L-LPS-induced cell apoptosis.