硝普钠对豚鼠耳蜗外毛细胞能动性影响的初步研究

目的研究硝普钠(sodium nitroprusside,SNP)对耳蜗OHC在电压刺激下的能动性的改变情况.方法运用全细胞膜片钳电压钳技术,在正常细胞内外液的条件下,观察不同浓度SNP对电压性刺激引起OHC能动性的影响情况.结果在SNP浓度低于100 μMol/L时,SNP对OHC能动性的影响不明显,但当SNP浓度高于100 μMol/L时,SNP对OHC的能动性有明显的抑制作用,结果有统计学意义(P<0.05).结论 SNP对OHC的能动性有抑制作用,作用呈一定的浓度效应关系.     

Longitudinal behaviour of neural response telemetry (NRT) data and clinical implications

Neural response telemetry (NRT) data from 63 subjects equipped with the Nucleus CI24M Cochlear Implant System generally exhibited little change over up to 4 years. Larger changes, when they occurred, were seen only within the first 15 months postoperatively, and these changes diminished over time. Intraoperative NRT data were generally stable enough to be used for assisting in the initial speech processor fitting sessions. It was not possible to predict changes in the subjective map threshold and comfortable loudness levels (T and C levels, respectively) based on observed changes in the NRT data. The long-term stability of the neural response amplitude and the neural response threshold, however, implies that NRT may be useful as a routine diagnostic tool to detect changes to the neural periphery over time.     

Development of atrial fibrillation in patients with atrioventricular block after atrioventricular synchronized pacing

Many studies have evidenced an increased incidence of AF in patients receiving single chamber ventricular pacing (VVI) when compared with those undergoing an atrial-based system (AAI or DDD). However, the difference in incidence of AF between two atrial-based systems (VDD, DDD) in patients with AV block was still controversial. This study was conducted to compare the development of AF between different modes of pacemakers (VDD and DDD) in patients with symptomatic AV block. A retrospective review was conducted of the detailed records of all consecutive patients who received permanent pacemakers due to symptomatic bradycardia from March 1995 to March 2000. The occurrence of AF was documented when there was presence of AF in the free-run or 12-lead ECG, any ECG strips, or persistent AF on 24-hour Holter ECG during the follow-up. The study included 152 patients (44 women, 108 men; mean age 73). The patients were divided into two groups: VDD (n = 100) and DDD (n = 52). The mean follow-up was 48.9 +/- 22.9 months. The incidence of AF was 7.9%. A higher incidence of AF was noted in the DDD group (15.4%) when compared with the VDD group (4.0%, P = 0.023). The incidence of development of AF in patients with AV block was higher in those receiving DDD cardiac pacing when compared with those who received the VDD system. The authors suggest that VDD pacing may be a better choice than the DDD system for patients with AV block, but without clinical evidence of sinus node dysfunction, and if an atrial lead is required, it should be placed close to the Bachmann's bundle.     

Detection of SARS coronavirus in patients with suspected SARS

Cases of severe acute respiratory syndrome (SARS) were investigated for SARS coronavirus (SARS-CoV) through RNA tests, serologic response, and viral culture. Of 537 specimens from patients in whom SARS was clinically diagnosed, 332 (60%) had SARS-CoV RNA in one or more clinical specimens, compared with 1 (0.3%) of 332 samples from controls. Of 417 patients with clinical SARS from whom paired serum samples were available, 92% had an antibody response. Rates of viral RNA positivity increased progressively and peaked at day 11 after onset of illness. Although viral RNA remained detectable in respiratory secretions and stool and urine specimens for >30 days in some patients, virus could not be cultured after week 3 of illness. Nasopharyngeal aspirates, throat swabs, or sputum samples were the most useful clinical specimens in the first 5 days of illness, but later in the illness viral RNA could be detected more readily in stool specimens.     

胶质瘤多药耐药细胞株大鼠动物模型的建立及其生物学性质

目的建立神经胶质瘤多药耐药细胞株C6/adr及其原始细胞株C6种植在S-D大鼠脑内的动物模型,对照研究其生物学性质.方法体外培养神经胶质瘤多药耐药细胞株C6/adr及其原始细胞株C6,在磁共振连续动态监测下,进行大鼠脑内接种,于接种当天、第3天以及接种后每周进行磁共振随访检查,并对同期大鼠进行组织活检,观察接种后肿瘤大小变化、荷瘤大鼠的生存期.结果体外培养的神经胶质瘤多药耐药细胞株接种大鼠4周以后出现明显的颅内压增高症状,磁共振阳性结果远早于症状的出现,能更准确地动态显示肿瘤在脑内的生长情况,接种后同期病理结果证实了磁共振影像学表现.C6和C6/adr细胞大鼠动物模型的生长特性相似,两者接种后同期的肿瘤大小、生存期无显著差异(P＞0.05).结论应用磁共振对大鼠动物模型进行动态监测,结合同期的病理切片,能够全面了解活体肿瘤的生长情况,并可确立大鼠脑内肿瘤生长的早、中、晚分期,为体内实验的进一步深入研究提供重要的工具和必要的基础.     

Helicobacter pylori eradication to prevent gastric cancer in a high-risk region of China: a randomized controlled trial

Context  Although chronic Helicobacter pylori infection is associated with gastric cancer, the effect of H pylori treatment on prevention of gastric cancer development in chronic carriers is unknown. Objective  To determine whether treatment of H pylori infection reduces the incidence of gastric cancer. Design, Setting, and Participants  Prospective, randomized, placebo-controlled, population-based primary prevention study of 1630 healthy carriers of H pylori infection from Fujian Province, China, recruited in July 1994 and followed up until January 2002. A total of 988 participants did not have precancerous lesions (gastric atrophy, intestinal metaplasia, or gastric dysplasia) on study entry. Intervention  Patients were randomly assigned to receive H pylori eradication treatment: a 2-week course of omeprazole, 20 mg, a combination product of amoxicillin and clavulanate potassium, 750 mg, and metronidazole, 400 mg, all twice daily (n = 817); or placebo (n = 813). Main Outcome Measures  The primary outcome measure was incidence of gastric cancer during follow-up, compared between H pylori eradication and placebo groups. The secondary outcome measure was incidence of gastric cancer in patients with or without precancerous lesions, compared between the 2 groups. Results  Among the 18 new cases of gastric cancers that developed, no overall reduction was observed in participants who received H pylori eradication treatment (n = 7) compared with those who did not (n = 11) (P = .33). In a subgroup of patients with no precancerous lesions on presentation, no patient developed gastric cancer during a follow-up of 7.5 years after H pylori eradication treatment compared with those who received placebo (0 vs 6; P = .02). Smoking (hazard ratio [HR], 6.2; 95% confidence interval [CI], 2.3-16.5; P<.001) and older age (HR, 1.10; 95% CI, 1.05-1.15; P<.001) were independent risk factors for the development of gastric cancer in this cohort. Conclusions  We found that the incidence of gastric cancer development at the population level was similar between participants receiving H pylori eradication treatment and those receiving placebo during a period of 7.5 years in a high-risk region of China. In the subgroup of H pylori carriers without precancerous lesions, eradication of H pylori significantly decreased the development of gastric cancer. Further studies to investigate the role of H pylori eradication in participants with precancerous lesions are warranted.      

Impact of cadmium exposure on male sex hormones: a population-based study in China

The objective of this study was to investigate the possible effects of environmental cadmium (Cd) exposure on the levels of serum sex hormones in a Chinese population group. A total of 263 male volunteers were included. Blood samples were collected for the determination of serum testosterone (T), measured by radioimmunoassay, and follicle stimulating hormone (FSH) and luteinizing hormone (LH), both measured by enzyme immunoassays. Urinary and blood Cd were analyzed by atomic absorption spectroscopy (AAS). We found a dose-response relationship between urinary Cd excretion and the prevalence of abnormally high serum T levels, but, through multiple regression analysis, we could not trace exposure to Cd as a significant determinant of serum T levels. Exposure to Cd also failed to influence the levels of FSH and LH in serum. In contrast, we found that age, body mass index (BMI), and smoking habits are significant determinants of FSH and LH and of T and LH, respectively. We conclude that oral Cd exposure is not a critical determinant of hormone homeostasis in males, but lifestyle and some biological factors, such as age and BMI, are important. The relationship found between urinary Cd and high T levels may be of importance for male reproductive morbidity and should be investigated further.     

P2X7 receptors stimulate AKT phosphorylation in astrocytes

1. Emerging evidence indicates that nucleotide receptors are widely expressed in the nervous system. Here, we present evidence that P2Y and P2X receptors, particularly the P2X(7) subtype, are coupled to the phosphoinositide 3-kinase (PI3K)/Akt pathway in astrocytes. 2. P2Y and P2X receptor agonists ATP, uridine 5'-triphosphate (UTP) and 2',3'-O-(4-benzoyl)-benzoyl ATP (BzATP) stimulated Akt phosphorylation in primary cultures of rat cortical astrocytes. BzATP induced Akt phosphorylation in a concentration- and time-dependent manner, similar to the effect of BzATP on Akt phosphorylation in 1321N1 astrocytoma cells stably transfected with the rat P2X(7) receptor. Activation was maximal at 5 - 10 min and was sustained for 60 min; the EC(50) for BzATP was approximately 50 microM. In rat cortical astrocytes, the positive effect of BzATP on Akt phosphorylation was independent of glutamate release. 3. The effect of BzATP on Akt phosphorylation in rat cortical astrocytes was significantly reduced by the P2X(7) receptor antagonist Brilliant Blue G and the P2X receptor antagonist iso-pyridoxal-5'-phosphate-6-azophenyl-2',4'-disulfonic acid, but was unaffected by trinitrophenyl-ATP, oxidized ATP, suramin and reactive blue 2. 4. Results with specific inhibitors of signal transduction pathways suggest that extracellular and intracellular calcium, PI3K and a Src family kinase are involved in the BzATP-induced Akt phosphorylation pathway. 5. In conclusion, our data indicate that stimulation of astrocytic P2X(7) receptors, as well as other P2 receptors, leads to Akt activation. Thus, signaling by nucleotide receptors in astrocytes may be important in several cellular downstream effects related to the Akt pathway, such as cell cycle and apoptosis regulation, protein synthesis, differentiation and glucose metabolism.     

Substance P induces inward current and regulates pacemaker currents through tachykinin NK1 receptor in cultured interstitial cells of Cajal of murine small intestine

We investigated whether substance P modulates pacemaker currents generated in cultured interstitial cells of Cajal of murine small intestine using whole cell patch-clamp techniques at 30 degrees C. Interstitial cells of Cajal generated spontaneous inward currents (pacemaker currents) at a holding potential of -70 mV. Tetrodotoxin, nifedipine, tetraethylammonium, 4-aminopyridine, or glibenclamide did not change the frequency and amplitude of pacemaker currents. However, divalent cations (Ni2+, Mn2+, Cd2+, and Co2+), nonselective cationic channel blockers (gadolinium and flufenamic acid), and a reduction of external Na+ from normal to 1 mM inhibited pacemaker currents indicating that nonselective cation channels are involved in their generation. Substance P depolarized the membrane potential in current clamp mode and produced tonic inward pacemaker currents with reduced frequency and amplitude in voltage clamp mode. [D-Arg1, D-Trp7,9, Leu11] substance P, a tachykinin NK1 receptor antagonist, blocked these substance P-induced responses. Furthermore, [Sar9, Met(O2)11] substance P, a specific tachykinin NK1 receptor agonist, depolarized the membrane and tonic inward currents mimicked those of substance P. Substance P continued to produce tonic inward currents in external Ca2+-free solution or in the presence of chelerythrine, a protein kinase C inhibitor. However, substance P-induced tonic inward currents were blocked by thapsigargin, a Ca2+-ATPase inhibitor in the endoplasmic reticulum or by an external 1 mM Na+ solution. Our results demonstrate that substance P may modulate intestinal motility by acting on the interstitial cells of Cajal by activating nonselective cation channels via the release of intracellular Ca2+ induced by tachykinin NK1 receptor stimulation.     

Development of a sensitive and HTS-compatible reporter gene assay for functional analysis of human adenosine A2a receptors in CHO-K1 cells

Adenosine A2a receptor, a member of the G protein-coupled receptor superfamily, has been demonstrated to be an important pharmacological target. It couples to stimulatory G protein and activates adenylate cyclase upon agonist stimulation. Here we attempted to stably transfect Chinese hamster ovary (CHO-K1) cells, which lack any known subtypes of adenosine receptors, with recombinant human adenosine A2a receptors (hA2aR). Rapid down-regulation of hA2aR in a clonal cell line, CHOA2a-2, was observed over a short period of time in culture. This is consistent with other groups' findings of low expression and poor G protein coupling of this receptor in several cell systems. To facilitate pharmacological profiling for hA2aR ligand, we introduced a cyclic AMP response element (CRE)-linked beta-galactosidase reporter gene into CHOA2a-2 cells to generate a stable cell line, CHOA2a-2CREbetagal#26. Robust cyclic AMP signal amplification was obtained using a colorimetric assay measuring beta-galactosidase activity. The EC(50) of 5'-N-ethylcarboxamidoadenosine (NECA), a potent A2a agonist, for inducing beta-galactosidase activity was 23.3 +/- 3.5 nM, similar to 22.7 +/- 3.9 nM, which was the NECA EC(50) in the direct measurement of cyclic AMP of CHOA2a-2 cells in early culture. Subsequently we validated this assay for high throughput screening for hA2aR agonists. The Z' factor for robotic assay performance was 0.79 +/- 0.03, the ratio of signal/noise was 157 +/- 36, and the ratio of signal/background was 10.6 +/- 1.2, demonstrating that this assay is well suitable for quality high throughput screening. High throughput screening of Johnson & Johnson libraries uncovered a couple of distinct series of nonadenosine small molecules, in addition to adenosine analogues, as potential hA2aR agonists with EC(50) values of 2-6 microM. Preliminary characterization of those compounds was presented.