Recent work has shown that nitric oxide (NO) induction by nitric oxide synthase(NOS) is the physiological mediator of bone cell function and demonstrated that it may be possible to exert differential effects on osteoblast (OB) and osteoclast (OC) activity in vivo. The proinflammatory cytokines, such as tumor necrosis factor-? (TNF-?), interleukin-1? (IL-1?), interferon-? (IFN-?), will stimulate bone resorption by NOS-induced low levels of NO. These findings confirm NO as a potentially important osteoblast-osteoclast coupling factor, indicating that cytokine-induced NO was largely responsible for the mechanisms of osteoporosis. Pharmacological modulation of NO may therefore represent a new approach in the treatment of bone diseases characterized by increased bone resorption, such as osteoporosis (OP).

Objective To analyze the influence of sleep deprivation on expression of serotonin receptor 1A(5-HT1A) and dopanine-2 receptor (D2R) gene and to explore the differences between different neurotransmitter pathways involved in sleep regulation through measuring the gene expression of 5-HT1A and D2R in regions of hippocampus,hypothalamus and striatum with different sleep deprivation models.Methods Sleep deprivation was performed to male SD rats of 10-week-old for 24 hours,48 hours and 72 hours respectively as the experimental group and a control group was taken for comparison.The expressions of 5-HT1A and D2R gene in regions of hippocampus,hypothalamus and striatum were detected through RT-PCR technique to analyze the influence of sleep deprivation on gene expression in different regions.Results Sleep deprivation had a significant effect on the gene expression of 5-HT1A in regions of hippocampus and striatum(F=56.203,P＜0.01 ; F=77.288,P＜0.01).The three experimental groups were all superior to the control group and the difference was of statistic significance(P＜0.05).In the hippocampus region,the expression quantity of the 72 hours group(0.618±0.054) was superior to that of the 24 hours group and of the 48 hours group(24 hours:0.404±0.023,P＜0.01 ;48 hours:0.455±0.042.P＜0.05).In the striatum region,the differences between the 24 hours group(0.413±0.033),the 48 hours group(0.464±0.034)and the 72 hours group(0.610±0.040) were all of statistic significance(all P＜0.05).Sleep deprivation had a significant effect on the expression of D2R gene in regions of hippocampus and striatum(F=74.708,P＜0.01 ; F=80.687,P＜0.01).The expression quantity of the three experimental groups in regions of hippocampus (24 hours:0.386±0.027,48 hours:0.318±0.014,72 hours:0.250±0.010) and striatum(24 hours:0.396±0.013,48 hours:0.349±0.017,72 hours:0.260±0.013) were all inferior to the control group.The differences were of statistic significance (all P＜0.05).There was a negative correlation between the gene expressions of 5-HT1A and D2R of rats of the three experience groups(all P＜0.05).Conclusion For the sleep deprivation rats,the gene expression of 5-HT1A rises while that of D2R falls in regions of hippocampus,hypothalamus,and there is a negative correlation between the expressions of the two genes.

Objective To investigate the role of Catbindin-D28k in the kidney on calcium metabolism.Methods VDR/CaBP-D28k double knockout(KO)mice was made.Body weight,diet intake and serum,urinary parameters and length,density of the long bones,histological staining of the tibia of WT,CaBP-D28(-/-),VDR(-/-)and VDR(-/-)/CaBP-D28k(-/-)mice were determined on regular and high Ca-Lac diet.Results On a regular diet,the double KO mice were growth-retarded more and smaller than VDR KO mice.Compared with VDR KO mice,the double KO mice had higher urinary calcium excretion and rachitic skeletal phenotype,which were manifested with higher serum parathyroid hormone levels,lower bone mineral density,and more distorted growth plate with mole osteoid formation in the trabecular region.On high calcium and high lactose diet,blood-ionized calcium levels were normal in both VDR KO and the double KO mice.However,in contrast to VDR KO mice,the skeletal abnormalities were not completely corrected in the double KO mice.Conclusion These results directly demonstrate that CaBP-D28k plays a critical role in maintaining calcium homeostasis and skeletal mineralization and suggest that its caleemic role can be mostly compensated by CaBP-D9k.

To clone and express 27 kDa cysteine protease (CP) gene of Spirometra erinacei plerocercoid,and analyze the biology characteristics,a total of RNA was extracted from the plerocercoids and reversely transcribed into cDNA.The 27kDaCP gene was amplified by PCR and cloned into pM-19T vector for sequencing.The accurate sequence was subcloned into the expression vector pET-28a (+).The recombinant plasmid was transformed into Transetta (DE3) and the expression protein induced by IPTG.The recombinant protein was purified by Ni2 + affinity chromatography,and analyzed by SDS-PAGE and Western blotting.The 27 kDa CP gene and its expression protein were predicted and analyzed by bioinformatics analysis tools such as NCBI and ExPASy.Results showed that the ORF length of 27 kDa CP gene sequence was 1 011 bp,and the removed signal peptide sequence was 954 bp with the submission number of ANA52569 in GenBank.The whole sequence of 27 kDa CP (Mr 35 669.9,pI 5.92) was 317 amino acids conferred from cDNA,which belongs to the Peptidase_C39_like superfamily.The pET-28a (+)-27kDa-CP was expressed under the induction of IPTG.Western blotting analysis showed that the purified protein reach expectancy,and had better response with positive serum of Spirometra erinacei plerocercoid infection.In conclusion,the 27 kDa CP gene of Spirometra erinacei plerocercoid is successfully cloned and expressed and knowing coded sequences and bioinformatic.

were obviously increased. Conclusion HIF-1α was successfully cloned. HIF-1α-pcDNA3.1 can be effectively transfected into MSCs with liposome-mediated method, which can result stable expression of HIF-1αin transfected MSCs.

OBJECTIVE To observe a time course of keratinocyte growth factor(KGF) and surfactant protein A(SPA) changes in rats with Pseudomonas aeruginosa pneumonia and to find out the significances of them.METHODS Specific pathogen-free male Sprague-Dawley rats were used to study.Standard P.aeruginosa strain ATCC 27853 was instilled into airway by the tracheal route to induce model of pneumonia.Samples of lung tissue and BALF were harvested before infection,and 6 h,9 h and 72 h after infection.Six rats per each point were sacrificed for harvesting samples.Expression of KGF protein in lungs was detected by Western blotting.Western-dot-blot was used to detect SPA expressions in BALF.RESULTS After infected with P.aeruginosa,KGF protein in lungs was markedly increased,reached the peak at 72 h postinfection.KGF protein level at 72 h postinfection was markedly higher than that of before infection(P

Objective To observe the immunoglobulin A secreting cells (IgASCs) and secretory IgA (sIgA) level in jejunum of mice infected with plerocercoids,and to understand the roles of resistance to invasion processes of the plerocercoids.Methods A total of 100 Kunming mice (half males and half females) were chosen,the weight was 20-25 g,they were randomly divided into control and experiment groups according to their body weight via the random number table method,50 per group.Mice of experiment group were fed each with 5 plerocercoids in snakes,and mice of control group were not infected,testing time and methods were the same in the two groups.Ten mice were randomly sacrificed from one group on days 1,7,14,28 and 56 after infection,to collect empty intestinal juice and jejunal segment.The immunohistochemical method was used to examine the quantity of IgASCs in jejunal mucosa,while enzyme-linked immunosorbent assay (ELISA) was applied to test the level of slgA of jejunal fluid.Results The IgASCs were distributed in lamina propria of the jejunal mucosa,and the percentage of positive IgASCs reached the peak value [(64.24 ± 0.60)％] at d 1 after infection in experiment group,then decreased,and it was lower than control group at d 14 [(41.98 ± 0.42)％ vs (43.52 ± 0.94)％,t =-4.727,P ＜ 0.01].The sIgA level reached the peak value [(22.05 ± 1.43) mg/L] at d 7 after infection in experiment group,then decreased,and there was no statistical significant difference between control and experiment groups at d 56 [(20.00 ± 0.42) mg/L vs (21.26 ± 2.59) mg/L,t =1.516,P ＞ 0.05].There was a positive correlation between the percentage of positive IgASCs in the jejunal mucosa and sIgA level in the jejunal fluid at d 7 after infection (r =0.663,P ＜ 0.01),and there was a negative correlation between them at d 14 after infection (r =-0.542,P ＜ 0.05).Conclusion The plerocercoids infection might induce high level expressions of IgASCs and sIgA,they show positive correlation at d 7 after infection.

ObjectiveTo introduce a new technique to create a pulmonary valve biorifice for reconstruction of right ventricular outflow tract in tetralogy of Fallot (TOF),and to summarize its initial clinical experience and therapeutic effect.MethodsThe new technique regarding reconstruction of right ventricular outflow tract with a pulmonary valve biorifice was used in a total of 53 TOF cases (the observation group).The conventional technique regarding reconstruction of right ventricular outflow tract was used in other 50 TOF cases (the control group).The clinical dates of all cases were reviewed retrospectively.ResultsThe ages,weights,cardiopulmonary bypass time,cardiac arrest time,as well as the post operation ventilation support time were not different significantly between two groups.Compared with the contrul group,patients from the observation group had shorter duration of ICU stay.After operation,in the observation group,only 2 cases had large amount of pleural effusion,1 case meddle,and 8 cases little amount of pleural effusion; whereas,in the control group,the corresponding numbers were 1,5 and 17,respectively.At the time point of 1 week after operation,all patients were rechecked by echocardiography,no pulmonary valve stenosis was found.Moderate pulmonary valve regurgitation was found in 8 cases,mild regurgitation in 15 cases from the observation group; and severe regurgitation in 3 cases,moderate regurgitation in 17 cases,and mild regurgitation in 16 cases from the control group.A total of 33 cases from the observation group were rechecked at the time point of half year after operation,and moderate - mild pulmonary regurgitation were found in 3 cases.A total of 18 cases of them were rechecked 1 - year latter,no pulmonary regurgitation was found.ConclusionsThe new technique to create pulmonary valve biorifice can reduce the pulmonary valve regurgitation and postoperative pleural effusion,and improve the early outcomc.

Objective To achieve the best chance and optimize the method of operation,the clinical outcomes of 76 cases with complete atrioventricular septal defect (CAVSD) were summarized.Methods According to the Rastelli classification,there were 57 cases of type A,6 type B,and 13 type C.The repaired procedures included the two-patch technique for atrioventricular septal defect (65 cases),direct closure of ventricular septal defect (7 cases),and the Glenn bidirection shunt (4 cases).Results Two patients died.Of them,one was concomitant with double outlet right ventricle (DORV) and total anomalous pulmonary venous connection (TAPVC),died of low cardiac output syndrome; another was complicated with severe pulmonary hypertension,and the death reason was hypoxaemia and respiratory function failure.The survived patients were followed up,and the follow-up period was varied from one to ten years,mitral valve regurgitation was found in 12 cases,3 were middle and 9 were mild.Conclusions In order to prevent deteriorated condition of these patients and improve the survival rate,CAVSD should be operated as soon as the diagnosis is certain,and the co-exist malformation also should be corrected.

Objective:To analyze gene expression profiling of left ventricular myocardium in patients with chronic congestive heart failure(CHF)caused by rheumatic heart disease(RHD)with the normal controls in order to identify CHF associated target genes. Methods:The gene expression profiles of left ventricular myocardium from patients with CHF by RHD and normal controls were obtained from six human whole-genomic oligonucleotide microarrays(Affymetrix HG U133 Plus 2.0 GeneChip). GeneSpring software was used to identify the differentially expressed genes in both groups,and bioinformatic analysis was applied to analyze the target genes associated with CHF. Real-time PCR was carried out to validate the expression of three target genes. Results:We identified 102 target genes associated with CHF which were classified into 7 gene clusters. Microarray results were further confirmed by real time PCR for three genes. ATF3 was markedly down-regulated,IGFBP2 and NPPB were notably up-regulated in the left ventricular myocardium samples from CHF patients. Conclusion:A lot of differentially expressed genes,obtained by using the whole-genomic expression profiling technology,might be a contributory factor for the initiation and progression of CHF and it helpful for the understanding of underlying pathophysiological implications. Further investigation on these genes would provide a strategy to identify genetic markers and molecular events associated with CHF caused by RHD.