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 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.

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

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.

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 determine the role of flagellar motor protein MotA in the pathogenesisassociated chemotaxis and colonization of Campylobacter jejuni. Methods The motA gene as well as Kan~r gene and plus-motA gene segments for motA gene knock-out were amplified by PCR and the target amplification fragments were sequenced after cloning. A suicide plasmid(pBlueskrit-Ⅱ-SK~(motA-kan)) and a motA gene knock-out mutant (motA~-) were constructed based on homologious recombination. By using semisolid plate migration test, hard agar plus (HAP)-based chemotactic test towards sodium deoxycholate (SDC) in vitro, and jejunal colonization test in BALB/c-ByJ mice were performed to determine the differences of flagellar motility, chemotaxis towards SDC and colonization in murine jejunum between motA~- mutant and wild-type strain. Results The nucleotide and amino acid sequences of the cloned motA gene were 100% identical to the reported corresponding sequences. The results of PCR, sequencing and continuous passage culture in antibiotics-contained medium demonstrated that both suicide plasmid and motA~- mutant were successfully generated. The diameters of clonies on semisolid plate and 0.2 mol/L SDC-induced chemotactic tings in HAP as well as the bacterial numbers adhering to the surface of murine jejunal mucosa and in jejunal content of motA~- mutant were significantly less than those of wild-type strain(P<0.05). Conclusion A motA gene knock-out mutant of C. jejuni was successfully constructed in this study, motA is an essential gene for flagellax motility, pathogenesis-associated chemotaxis and colonization of C. jejuni.

BACKGROUND: Studies have demonstrated that exogenous basic fibroblast growth factor (bFGF) has intensive effects to promote proliferation of gingival fibroblasts (GFs) cultured in vitro and the heeling of gingival wounds. OBJECTIVE: To investigate the effects of bFGF gene transfection on the biological performance of Beagle canine GFs. DESIGN, TIME AND SETTING: An observation and comparison in vitro experiment regarding cells was accomplished in Centre of Cell Biology and Development of Fujian Medical University and Department of Comparative Medicine in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from April to September of 2008. MATERIALS: Four beagle dogs, male, 12 months old, weighing 10-13 kg were used in this experiment, plRES2-EGFP-bFGF plasmid containing full-length human bFGF gene cDNA was constructed and conserved by our institution. METHODS: Free gingiva of the 2nd, 3rd and 4th premolars were excised from left upper jaw of Beagle dogs, dnsed with aseptic phosphate buffer four times, then cut into pieces and digested with 2.5 g/L pancreetin for 2 hours at 37 ℃. After the cantrifugation and supernatant removal, DMEM containing 10% fetal bovine serum was added to incubate on 6-well plate with coverlips in 5% CO2 incubator at 37 ℃. Logadthrnically growing cells were digested and passaged. GFs were transfected with pIRES2-EGFP-bFGF plasmid using liposome mediated method, while vacant plasmid transfection and un-transfection group served as controls.MAIN OUTCOME MEASURES: Proliferation and apoptosis feature of the GFs were evaluated by M'rE and AOEB, respectively. The activity of alkaline phosphatase was assayed by chemical coledmetry. RESULTS: All of three groups cells entered log phrase on three days after transfection. MTT results showed that the proliferation of GFs transfected with bFGF was greater than cells transfected with vacant vector and untransfected cells (P < 0.05). AO/EB dyeing showed the apoptosis rate of GFs transfected with bFGF was reduced compared with other two groups (P < 0.05). After bGFG gene transfection, the ALP activity remained unchanged and there was no significant difference compared with untransfected cells.CONCLUSION: The transfection of bFGF gene to GFs can promote the proliferation of GFs and depress the apoptosis. No promotion is present with regard to the GFs differentiation.

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.

Objectiye To study the mechanism by which transcranial magnetic stimulation (rTMS) affects cognitive dysfunction in vascular dementia (VD). Methods Thirty-six male Wistar rats were randomly divided into a control group, a VD group, a low frequency rTMS group and a high frequency rTMS group. Two-vessel occlusion was employed to induce VD models. Low frequency rTMS group rats were given 0.5 Hz rTMS for six weeks. High frequency rTMS group rats were given 5 Hz rTMS for six weeks. Morris' water maze test was used to measure their spatial learning ability and memory. The ultrastructures of the synapses in the four groups were detected with transmission electron microscopy. The expression of synaptophysin (SYN), brain derived neurotrophic factor (BDNF) and Nmethyl-D-aspartate receptor 1 ( NMDAR1 ) mRNA and protein in the hippocampus were determined by real-time PCR and Western blotting. Results The behavior and morphology of the rats treated with rTMS improved. The average expression of SYN, BDNF and NMDAR1 mRNA and protein in the low frequency rTMS group and the high frequency rTMS group were significantly higher than in the VD group. Conclusion rTMS can provide a rehabilitative effect for VD. The mechanism might be associated with enhancing the expression of SYN, BDNF and NMDAR1 in the hippocampus.