荧光原位杂交技术分析人结肠菌群方法研究

建立荧光原位杂交技术分析人体内结肠菌群的方法。取受试者新鲜粪便 ,选用 5种特异性的 16SrRNA寡核苷酸探针 ,检测粪便样本收集后的保存时间、温度 ,离心条件及样本固定液存放时间对杂交计数结果的影响。结果建立最佳实验条件为 :粪便样本收集后应尽快在 4℃下保存 ,放置时间不要超过 12小时即作处理 ;样本的适宜离心条件为 70 0g 2分钟 ;样本用多聚甲醛固定后在 - 80℃下存放时间不要超过 5个月。该方法具有较好的稳定性 ,可以有效地检出个体之间结肠菌群的差异。     

Diffusion-weighted magnetic resonance imaging for demonstration of cerebral infarcts

Summary Forty-one patients demonstrating clinical symptoms for cerebral infarction were investigated by magnetic resonance imaging with diffusion-weighted echo-planar imaging (DWI) and T2-weighted imaging (T2WI). In 8 patients only DWI showed the cerebral lesions clearly. One patient with positive DWI and T2WI suffered from HSV encephalitis. DWI is superior to T2WI in assessment of small cortical infarcts and cerebral infarction in patients with preexisting vascular lesions. DWI is not specific, so other causes like cerebral hematoma and encephalitis have to be considered.      

Do callosal projection neurons reflect sex differences in axon number?

We have reported that female rats have more axons in the splenium of the corpus callosum than do male rats (12). To determine if the greater number of axons found in female rats might be reflected in a larger distribution of callosal projection neurons, horseradish peroxidase (HRP) was injected into the visual cortex of 55-65-day-old rats of both sexes that had been housed in a complex environment since weaning. The pattern of labeled neurons was examined in tangential sections in the cortex contralateral to the injection site, and three-dimensional reconstructions were quantified at the area 17/18a border and in area 18b. Male and female rats were found to have indistinguishable distributions of labeled callosal projection neurons. The present study failed to find an obvious difference in the distribution of projection neurons as the basis for the sex differences in axon number, but because of the limitations of tracing techniques, subtle differences cannot be excluded.     

L-arginine induces relaxation of small mesenteric arteries from endotoxin-treated rats.

The effect of arginine (Arg) was studied on norepinephrine- (1 microM) precontracted small mesenteric arteries removed from rats treated with E. coli lipopolysaccharide (LPS). The addition of L- (but not D-) Arg (1 mM) relaxed, within 3 min, the small mesenteric arteries from LPS-treated but not from control rats, the maximal relaxation (65.3 +/- 11%) being reached with less than 100 microM L-Arg. NG-Nitro-L-arginine methyl ester (1 mM) and methylene blue (10 microM) restored contractions to the level reached before addition of L-Arg. These results show that LPS induces the production of an L-Arg-derived, nitric oxide-like, relaxing factor in small mesenteric arteries.     

Improved anchorage in osteoporotic vertebrae with new implant designs.

The goal of our study was to evaluate two newly developed implant designs and their behavior in terms of subsidence in lumbar vertebral bodies under cyclic loading. The new implants were evaluated in two different configurations (two small prototypes vs. one large prototype with similar load-bearing area) in comparison to a conventional screw-based implant (MACS TL). A pool of 13 spines with a total of 65 vertebrae was used to establish five testing groups of similar bone mineral density (BMD) distribution with eight lumbar vertebrae each. In additional to BMD assessment via dual-energy X-ray absorptiometry, cancellous BMD and structural parameters were determined using a new generation in vivo 3D-pQCT. The specimens were loaded sinusoidally in force control at 1 Hz for 1000 cycles at three load levels (100, 200, and 400 N). A survival analysis using the number of cycles until failure (Cox regression with covariates) was applied to reveal differences between implant groups. All new prototype configurations except the large cylinder survived significantly longer than the control group. The number of cycles until failure was significantly correlated with the structural parameter Tb.Sp. and similarly with the cancellous BMD for three of five implants. In both large prototypes the cycle number until failure significantly correlated with the preoperative distance to the upper endplates. Although the direct relationship between bone structure or density and mechanical breakage behavior cannot be conclusively proven, all the prototypes adapted for poor bone structure performed better than the comparable conventional implant.     

Cardiac atrophy in women following bed rest

Both chronic microgravity exposure and long‐duration bed rest induce cardiac atrophy, which leads to reduced standing stroke volume and orthostatic intolerance. However, despite the fact that women appear to be more susceptible to post‐spaceflight presyncope and orthostatic hypotension than male astronauts, most previous high‐resolution studies of cardiac morphology following microgravity have been performed only in men. Because female athletes have less physiological hypertrophy than male athletes, we reasoned that they also might have altered physiological cardiac atrophy after bed rest. Magnetic resonance imaging was performed in 24 healthy young women (32.1±4 years) to measure left ventricular (LV) and right ventricular (RV) mass, volumes, and morphology accurately before and after 60 days of 6° head‐down tilt (HDT) bed rest. Subjects were matched and then randomly assigned to sedentary bed rest (controls, n=8) or two treatment groups consisting of (1) exercise training using supine treadmill running within lower body negative pressure plus resistive training (n=8), or (2) protein (0.45 g/kg/day increase) plus branched‐chain amino acid (BCAA) (7.2 g/day) supplementation (n=8). After sedentary bed rest without nutritional supplementation, there were significant reductions in LV (96±26–77±25 mL; P=0.03) and RV volumes (104±33–86±25 mL; P=0.02), LV (2.2±0.2–2.0±0.2 g/kg; P=0.003) and RV masses (0.8±0.1–0.6±0.1 g/kg; P<0.001), and the length of the major axis of the LV (90±6–84±7 mm. P<0.001), similar to what has been observed previously in men (8.0%; Perhonen M.A., Franco F., Lane L.D., Buckey J.C., Blomqvist Zerwekh J.E., Peshock R.M., Weatherall P.T., Levine B.D. J Appl Physiol 2001: 91: 645–653). In contrast, there were no significant reductions in LV or RV volumes in the exercise‐trained group, and the length of the major axis was preserved. Moreover, there were significant increases in LV (1.9±0.4–2.3±0.3 g/kg; P<0.001) and RV masses (0.7±0.1–0.8±0.2 g/kg; P=0.002), as well as mean wall thickness (9±2–11±1 mm; P=0.02). The interaction between sedentary and exercise LV and RV masses was highly significant (P<0.0001). Protein and BCAA supplementation led to an intermediate phenotype with no change in LV or RV mass after bed rest, but there remained a significant reduction in LV volume (103±14–80±16 mL; P=0.02) and major‐axis length (91±5–88±7 mm; P=0.003). All subjects lost an equivalent amount of body mass (3.4±0.2 kg control; 3.1±0.04 kg exercise; 2.8±0.1 kg protein). Cardiac atrophy occurs in women similar to men following sedentary 60 days HDT bed rest. However, exercise training and, to a lesser extent, protein supplementation may be potential countermeasures to the cardiac atrophy associated with chronic unloading conditions such as in spaceflight and prolonged bed rest.     

Alteration of visual evoked potentials and electroretinograms in Parkinson's disease

A group of 24 patients with Parkinson's disease (PD) with normal fundi and normal visual acuities was examined electrophysiologically. Checkerboard reversal VEPs and ERGs (P-ERGs) at various contrast levels as well as photopic and scotopic luminance ERGs were recorded and compared with an age-matched group of controls. Earlier reported latency increases of the VEPs of the patients were confirmed for patterns of high contrast only. Scotopic and photopic luminance ERGs of the patients showed normal latencies, but at all light intensities the amplitudes of the scotopic and photopic b wave, as well as the amplitudes of the photopic a waves, were significantly reduced, P-ERG amplitudes were reduced at 50% contrast. Identical results were obtained in patients under dopaminergic treatment (n = 17) and in patients who did not receive any treatment (n = 7). These results suggest that alterations occur already at the retinal level where dopamine receptors have been found. Thus the reported changes of the VEP are not caused by the visual cortex alone.     

Antigenic structure of the hexacosapeptide melittin: evidence for three determinants, one with a helical conformation.

ELISA-based epitope analysis was performed using rabbit polyclonal antisera against melittin. Antigenic sites were found at the C-terminus, in the middle section and within the N-terminal helix. Antibodies against the helical segment could discriminate between two faces of the amphiphilic helix. The antigenic sites include the bulk of the melittin hexacosapeptide, which is synonymous with a very high epitope density.