Experiments were performed on in vitro brainstem slice preparations from neonatal rats that retain respiratory network activity. Extracellular recordings were made from 99 neuronal units, respiratory-related or non-respiratory related with rhythmical activity, out of which there were 7 biphasic expiratory and 11 inspiratory ones. Possible roles of non-NMDA receptors in reciprocal excitation among the biphasic expiratory neurons and in excitatory synaptic inputs to inspiratory neurons were investigated by administration of non-NMDA receptor agonist KA and its antagonist DNQX in the perfusion solution. Bath application of non-NMDA receptor agonist KA increased the peak frequency of both biphasic expiratory and inspiratory neuronal discharges, and increased the discharge frequency of the biphasic expiratory neurons and the inspiratory neurons in the middle phase, while the frequency of discharge in the early and late phases were less affected. All of these effects were blocked by addition of the non-NMDA receptor antagonist DNQX, suggesting the involvement of non-NMDA receptors.

Objective To study the role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the generation and modulation of basic respiratory rhythm. Methods Respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve was recorded by suction electrode on the brainstem slices isolated from the neonatal rats, and the effects of the excitatory amino acids and its antagonists on the RRDA were investigated by adding these drugs into the modified Kreb's solution perfusing the brainstem slices. Results After application of the non-NMDA receptors agonist KA, it was found that the respiratory cycle and the expiratory time were slightly lengthened, but the NMDA receptor agonist NMDA had no effect on the RRDA. Both of the mutual antagonist DNQX and AP5 remarkably decreased the discharge frequency and the integral amplitude, accompanied by the shortening of the inspiratory time; DNQA simultaneously shortened the respiratory cycle and the expiratory time. Conclusion During the generation and the modulation of the mammalian respiratory rhythm, NMDA receptors act mainly to regulate the amplitude of the respiratory activity, and the non-NMDA receptors can not only affect the respiratory amplitude but also modulate the respiratory rhythm.

Objective To study the role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the generation and modulation of basic respiratory rhythm. Methods Respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve was recorded by suction electrode on the brainstem slices isolated from the neonatal rats, and the effects of the excitatory amino acids and its antagonists on the RRDA were investigated by adding these drugs into the modified Kreb's solution perfusing the brainstem slices. Results After application of the non-NMDA receptors agonist KA, it was found that the respiratory cycle and the expiratory time were slightly lengthened, but the NMDA receptor agonist NMDA had no effect on the RRDA. Both of the mutual antagonist DNQX and AP5 remarkably decreased the discharge frequency and the integral amplitude, accompanied by the shortening of the inspiratory time; DNQA simultaneously shortened the respiratory cycle and the expiratory time. Conclusion During the generation and the modulation of the mammalian respiratory rhythm, NMDA receptors act mainly to regulate the amplitude of the respiratory activity, and the non-NMDA receptors can not only affect the respiratory amplitude but also modulate the respiratory rhythm.

OBJECTIVETo understand the role of KCNE2 in functional regulation of Kv4.3, the major alpha subunit of transient outward current (I(to)) in human heart.

METHODSThe cDNAs of Kv4.3 or Kv4.3 plus KCNE2 were transfected into COS-7 cells and 24-36 h after the transfection, the channel proteins were expressed in the surface membrane of the cells and the channel currents were recorded with patch-clamp technique in whole-cell mode.

RESULTSKCNE2 played an important role in modulating the channel function. The recorded current density was decreased in cells co-expressing KCNE2 and Kv4.3 to 152.96-/+33.71 pA/pF (n=16) as compared with Kv4.3-expressing cells with a mean current density of 375.13-/+112.87 pA/pF (n=11). At the recording voltage of 60 mV, KCNE2 increased the time to peak (TTP) of the current. TTP in only Kv4.3-expressing cells was 4.82-/+0.32 ms (n=11), significantly shorter than the TTP of 20.41-/+2.13 ms (n=16) in cells co-expressing Kv4.3 and KCNE2 (P<0.05). In the presence of KCNE2, the voltage-dependent inactivation of Kv4.3 showed a positive shift. The voltage of half maximum inactivation (V(0.5)) was decreased significantly from -53.62-/+1.24 mV (n=8) in Kv4.3 group to -46.58-/+1.6 mV (n=10) in KCNE2 co-expression group (P<0.05). KCNE2 accelerated the recovery of the channel from inactivation, reducing the recovery time constant (tau) from 193.43-/+17.98 ms to 137.71-/+18.29 ms.

CONCLUSIONKCNE2 might serve as an important beta subunit and play a role in the regulation of I(to) function in human heart.

Animals ; COS Cells ; Cercopithecus aethiops ; Humans ; Kv Channel-Interacting Proteins ; genetics ; metabolism ; Membrane Potentials ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; genetics ; metabolism ; physiology ; Shal Potassium Channels ; genetics ; metabolism ; physiology ; Transfection

The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

OBJECTIVETo explore the effect of donor-derived NK cells added to pretreatment conditioning regimen on hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.

METHODSMurine model of MHC haplotype-mismatched BMT was established by using BALB/c(H-2d) x C57BL/6(H-2b) (CB6F(1)(H-2d/b)) mouse as recipient, and C57BL/6(H-2b) mouse as donor. Fifty recipient mice were divided into 5 groups. The mice in the first three groups were each infused 1 x 10(6), 5 x 10(5), 2 x 10(5)/mouse donor-derived NK cells, respectively before TBI ((60)Co, 9.0 Gy) and then conditioned with TBI, followed by infusion of C57BL/6(H-2b) mice bone marrow cells four hours later. The mice in the fourth group received TBI only, and in the fifth group, TBI and BMT at the some doses as the first three groups. Hematopoietic reconstitution, survival time, body weight, histopathology of the recipients were followed up.

RESULTS(1) Survival time was (5.15 +/- 0.66) days for the fourth group, and > 30 days for the other 4 groups. (2) Leukocyte and platelet counts at day 10 after BMT were (0.99 +/- 0.22) x 10(9)/L and (61.0 +/- 7.27) x 10(9)/L respectively for the fifth group and (2.01 +/- 0.21) x 10(9)/L, (101.50 +/- 16.34) x 10(9)/L; (1.98 +/- 0.29) x 10(9)/L, (99.50 +/- 16.41) x 10(9)/L and (1.97 +/- 0.21) x 10(9)/L, (98.0 +/- 16.19) x 10(9)/L for the first three groups, respectively. Histopathology displayed no GVHD in all the groups.

CONCLUSIONDonor-derived NK cells could promote hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.

Animals ; Bone Marrow Transplantation ; Female ; Graft Survival ; Graft vs Host Disease ; immunology ; prevention & control ; Haplotypes ; Histocompatibility Testing ; Killer Cells, Natural ; cytology ; immunology ; Lymphocyte Transfusion ; Major Histocompatibility Complex ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transplantation Conditioning ; methods ; Transplantation, Homologous

Accidents, Occupational ; statistics & numerical data ; China ; epidemiology ; Humans

After the connotation and necessity of social capital participating in health information construction were analyzed, the principal foreign experiences, and domestic status quo and problems of social capital participating in health information construction were described and the following development strategies were put forward for the par-ticipation of social capital in health information construction, namely the key points for the participation of social capital in health information construction, smooth and effective management mechanisms, standard management systems and information security management systems, comprehensive and practical assessment and monitoring sys-tems, encouragement of the participation of social capital in health information construction.

OBJECTIVETo study the alteration of protein Z (PZ) in patients with cardio-cerebral thrombotic diseases, its clinical significance and relations with FX.

METHODSPZ and FX:Ag were measured by ELISA, and plasma FX:C by first stage method. In 170 patients with acute ischemic stroke (AIS), 40 acute myocardial infarction (AMI) and 60 healthy adults as contrast, PZ, FX:C and FX:Ag were measured and compared between incipience and recurrence, different ages and genders.

RESULTSIn AIS and AMI groups, PZ levels decreased significantly to (940.02 +/- 229.82) microg/L and (1071.44 +/- 180.52) microg/L, respectively \[the contrast group was (2257.97 +/- 479.76) microg/L, P < 0.001\]. But FX:C and FX:Ag raised to (136.73 +/- 34.93)% and (135.54 +/- 54.39)% in AIS group; and to (139.53 +/- 29.18)%, (129.75 +/- 21.91)% in AMI group, respectively, while in the contrast group they were (94.33 +/- 22.00)% and (77.22 +/- 13.19)% (P < 0.001). In the comparative research between the AIS group, AMI group and the contrast group, PZ level was clearly found to negatively relate to the level of FX:C and FX:Ag (P < 0.001). Meanwhile, PZ level, FX:C and FX:Ag in recur-AIS group and recur-AMI group exhibited significant differences (P < 0.05) from those in the primary AIS and AMI groups, suggesting that the decrease of PZ levels reflected the pathological process of the disease. In addition, PZ level gradually decreased with the increase of age (P < 0.05), while FX:C and FX:Ag had no relations with age (P > 0.05). No correlation was found in sex with PZ level, FX:C, FX:Ag (P > 0.05).

CONCLUSIONPZ level was significantly decreased in AIS and AMI patients and was negatively related to FX:C and FX:Ag. The mechanism leading to FX increase may partially related with the decreased of PZ. PZ level was different in the primary and recurrent disease and was gradually decreased with the increase of age. Lack of PZ might be a etiological factor of cardio-cerebral thrombotic diseases.

Aged ; Aged, 80 and over ; Blood Proteins ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Factor X ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; Stroke ; blood

BACKGROUND & OBJECTIVE: Except for the tight correlation to nasopharyngeal carcinoma, accumulating evidences show that EpsteinBarr virus (EBV) is correlated to other carcinomas. This study was to investigate the correlation of EBV to colorectal carcinoma in Chinese.METHODS: EBV DNA was detected by polymerase chain reaction (PCR) in 90 specimens of primary colorectal carcinoma and 25 specimens of corresponding adjacent non-cancerous tissue, with the primers covering 2 different regions of EBV genome, BamH I W fragment and latent membrane protein 1 (LMP1) exon 3. The expression of EBV nuclear antigen 1 (EBNA1) and LMP1 were determined by immunohistochemistry, and the expression of EBV-encoded RNAs (EBERs) was detected by in situ hybridization.RESULTS: EBV LMP1 exon 3 and W fragment were detected in 27.7% and 32.2% of the 90 colorectal carcinoma specimens, which were significantly higher than the positive rate of EBV gene in the 25 adjacent non-cancerous tissues (4.0%, P＜0.001). Of the 29 W fragment-positive tumors, 23 (79.3%) were EBNA1-positive, 1 (3.4%) was EBERs-positive; most EBNA1-positive cells were tumor cells with positive signals gathered in the nuclei. No expression of EBNA1 and EBERs were detected in the 8 W fragmentnegative tumors. No expression of LMP1 was detected in all tumor specimens. CONCLUSION: EBV infection might be associated with colorectal carcinoma in Chinese.