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.

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 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.

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.

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

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.

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.

ObjectiveTo explore and analyze the distribution of NAT2 gene in Han population in Shenzhen andprovidethebasisfortreatmentof NAT2-relatedmetabolicdiseasesandcancer research. MethodThe study diluted the DNA sample(d0 ng/μl) as follows: 1 × 100, 1 × 101, 1 × 102, 1 ×103 , 1 × 104, to verify the sensitivity of detection of NAT2 gene polymorphism by real-time PCR. Mutation locus of 282,341,481, 590 and 857 of NAT2 in 554 normal samples were detected and genotyped by realtime PCR with Taqman probes. Forty-seven cases among 554 healthy controls were detected by DNA sequencing to verify the sensitivity, specificity and accuracy of this assay. ResultsThe lowest detection limit was 10-4 ng/μl. The frequencies of each allele of NAT2 were: *4 allele 64. 6％ (358/554), *5 allele 6. 3％ (35/554), *6 allele 25. 3％ ( 140/554), *7 allele 30. 0％ ( 166/554), * 11 allele 0. 6％ (3/554) The frequencies of main genotypes of NAT2 were as the follows : NAT2 *4/* 6 12.3％ ( 68/554 ), *6/* 78. 3％ (46/554), * 13/* 13 or * 12/* 12 or *4/* 4 7.6％ (42/554), *4/* 7 8. 1％ (45/554), * 7/* 1 312. 3％ (68/554), * 12 7.2％ (40/554), *6/* 13( * 12)5.6％ (31/554). The samples with 7 genotypes account for 89. 6％ (496/554) of the total cases.Rapid acetylation and Intermediate type accounting for 40. 5％ (224/554) and 46. 7％ (259/554) respectively were the main phenotypes of NAT2 in Han people in Shenzhen. In addition, the accuracy, specificity and sensitivity of the PCR were compared to direct DNA sequencing. The accuracy of 282, 341,481, 590 and 857 were 88.2％ (30/34), 87.5％ (7/8), 80. 0％(4/5), 100. 0％ ( 22/22 ) and 93.8％ ( 15/16 ) respectively. The specificities were 100. 0％ ( 13/13 ),94. 9％ ( 37/39), 100. 0％ ( 42/42 ) , 96.0％ ( 24/25 ) and 96. 8％ ( 30/31 ) respectively. The sensitivities were 91.5％ ( 43/47 ), 93.6％ ( 44/47 ), 97.6％ ( 46/47 ) 97.9％ ( 46/47 ) and 95.7％ ( 45/47 ),respectively. ConclusionsThe sensitivity, specificity and accuracy of real-time PCR established in this study is good. It can be widely used for clinical and scientific research. The major alleles of NAT2 of Han population in Shenzhen are * 4, *6, * 7. The most common slow acetylation genotype is *6/* 7. This study can provide a basis for NAT2-related metabolic diseases and cancer research.

Objective To explore the influence of different injection techniques on the quality of bolus in 99mTc-DTPA renal dynamic imaging.Methods 395 patients accepted 99mTc-DTPA renal dynamic imaging were retrospectively analyzed.All patients were divided into three groups according to injection techniques:direct injection group (187 cases),intravenous route injection group (84 cases)and venous indwelling needle injection group (124 cases).The three groups were injected by each technique.Areas of interest (ROI) were drawn on abdominal aorta by Xeleris workstation in blood flow perfusion imaging.The time-radioactivity curves of ROI were got.The patients whose ROI curve formed a peak was successfully injected,and did not formed was unsuccessfully injected.The number of patients in three groups who were successfully or unsuccessfully injected was respectively calculated.The data of three groups was taken Chisquare test by SPSS17.0 software.Results 174 patients of the direct injection group,46 of the intravenous route injection group and 115 of the venous indwelling needle injection group were injected successfully.The successful rate respectively was 93.0％,54.8％ and 92.7％.The successful rate of the direct injection group and venous indwelling needle injection group were higher than intravenous route injection group.The difference had statistical significance.The successful rate of the direct injection group and venous indwelling needle injection group hadn't statistical significance.Conclusions The successful rates of the direct injection group and venous indwelling needle injection group were similar.The venous indwelling needle injection technique can be chosen.The successful rate of the intravenous route injection group was lower than the other two groups.The intravenous route injection technique should be chosen prudently.