Simultaneous detection of 7 important Rickettsiales pathogens by TaqMan-probe quantitative real-time PCR / 中华流行病学杂志

Objective: To establish and optimize a TaqMan-probe quantitative real-time PCR (qPCR) assay for the detection of 7 important Rickettsiales pathogens and simultaneous identification of the infection types. Methods: Based on the ompB gene of Rickettsia prowazekii, Rickettsia mooseri and spotted fever group rickettsiae, the groEL gene of Orientia tsutsugamushi, the 16S rRNA of Ehrlichia chaffeensis, the gltA gene of Anaplasma phagocytophilum and the com1 gene of Coxiella burnetii, we synthesized primers and TaqMan-probes and optimized the reaction system and reaction process to same solution. The sensitivity, specificity and reproducibility of this assay were evaluated and the assay was used for the detection of simulated and actual samples. Results: The Ct value of the standard curves of the 7 pathogens showed a good linear relationship with the number of DNA copies (all R2 >0.990 0), the minimum detection limit was 10 copies/μl, showing good specificity. In the 96 tick nucleic acid extracts, Coxiella burnetii was detected in 1 sampleand spotted fever group Rickettsiae was detected in 3 samples. In the 80 blood samples from patients with undefined febrile illness, Orientia tsutsugamushi was detected in 1 sample and spotted fever group rickettsiae was detected in 2 samples. Conclusions: In this study, based on the established TaqMan-probe qPCR assay, the reaction system and reaction condition of the 7 important pathogens of Rickettsiales were optimized to the same solution. This method overcomes the shortcomings of using different reaction systems and reaction conditions for different pathogens, which can precisely identify the species of 7 important pathogens of Rickettsiales in clinical sample detections and is important for the infection type identification and laboratory detection time reduction to facilitate precise treatment of the patients.

Effect of orexin-A and orexin-1 receptor antagonist injected into the fourth ventricle of rats on food-intake and spontaneous physical activity / 生理学报

The present study was aimed to investigate the effects of orexin-A and orexin-1 receptor (OX1R) antagonist injected into the fourth ventricle of rats on food-intake and spontaneous physical activity (SPA). Obese rat model was induced by high fat diet. Different doses of orexin-A or SB334867, an OX1R antagonist, were injected into the fourth ventricle of obese and normal rats respectively. SPA and food intake were monitored for 4 h after injection in both light and dark environment. In the light measurement cycle, different doses of orexin-A significantly stimulated feeding and SPA in all injected rats, and the animals' responses showed a dose-dependent manner (P < 0.05-0.01), and compared with those of normal rats, the orexin-A induced food intake and SPA were more pronounced in obese rats. In the dark measurement cycle, different doses of orexin-A had no obvious effect on food intake and SPA in both normal and obese rats (P > 0.05). In the light cycle, different doses of SB334867 significantly decreased food intake and SPA in all rats during 0-2 h and 2-4 h after injection (P < 0.05), but the food intake and SPA in obese rats were significantly greater than those of normal rats. In the dark cycle, different doses of SB334867 showed no obvious effect on food intake and SPA of normal and obese rats (P > 0.05). These results suggest that fourth cerebral ventricle nuclei may be one target for orexin-A and light condition may play an important role in orexin-A and OX1R physiological functional processes.

CT perfusion imaging study of the hemodynamics in patients with intracerebral hemorrhage / 中华神经医学杂志

Objective To investigate the hemodynamic changes in patients with intracerebral hemorrhage (ICH) using CT perfusion (CTP) imaging and the relation between cerebral perfusion and the Scandinavian Stroke Scale (SSS) of the patients. Methods Forty-one patients with supratentorial ICH underwent plain CT scanning and CTP imaging at different times after ICH onset (from 5 h to 14 days). The impact of the time after ICH onset, hematoma volume, edema area, mean hlood pressure (MBP) and blood glucose on the hemodynamics of the patients was assessed, and the correlation between the SSS and the hemodynamic changes evaluated. Resnlts The cerebral blood flow in the edema area (CBF1), the peripheral area of the edema (CBF2, within 1 cm to the edema) and the distant cortical area from the hematoma (CBF3) showed significant differences (r=0.027, P=0.870) and fluctuated with time. CBF1 showed a positive linear relation with CBF2 (r=0.334, P=0.035), but neither of them was correlated to CBF3 (r=0.027, P=0.870;r=0.142, P=0.383). CBF1 also showed positive linear relations with relative cerebral blood volume (rCBV1) (r=0.803,P=0.000) and the peak time (PT1) (r=-0.52, P=0.752). The reduction of CBF1 was inversely correlated to the hematoma volume and the edema area (r=-0.501, P=0.001;r=-0.589, P=0.000), but not related with MBP or blood glucose (r=0.141, P=0.386;r=0.014, P=0.930). The area of ischemic injury (the ischemic area defined by CBF parameters-the hematoma area, edema area (r=0.449, P=0.003;r=0.645, P=0.000), but not to the MBP or blood glucose (r=-0.047, P=0.769;r=0.141,P=0.378). SSS was found to correlate to the volume and location of the hematoma and the reduction of CBF and CBV (r=-0.418, P=0.007;r=0.542, P=0.000;r=0.376, P=0.017;r=0.312, P=0.050), but not to the ischemic and edema area (r=-0.283, P=0.073;r=-0.163, P=0.308). Conclusion CBF is reduced in the edema area, peripheral area of the edema and the distant cortical area from the hematoma but showing different patterns of variation. Ischemic injury is present in the edema area around the hemotema, and its scope and severity is related with the hemotoma volume and the size of the edema area and may vary with time. The location and volume of the hematoma as well as the perfusion level in the edema area all affect the SSS of the patients.

Expression of motilin and its precursor mRNA in normal parenchyma, benign and malignant tumors of human thyroid / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of motilin and its precursor mRNA in normal human thyroid. To compare the expression differences of motilin and it precursor mRNA between normal thyroid and intestines. To study the expression of motilin and its precursor mRNA in human thyroid tumors and their clinical implications.</p><p><b>METHODS</b>RT-PCR, Southern blot and molecular cloning were used to detect motilin transcript expression in human thyroid and mucous membrane of small intestine. Real-time PCR and immunohistochemical techniques were used to quantify motilin precursor mRNA and motilin peptide in thyroid tissue samples including adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter.</p><p><b>RESULTS</b>(1) The expression of motilin and its precursor mRNA in normal human thyroid was primarily in the thyroid C cells. (2) RT-PCR and Southern blot showed that motilin mRNA expressed in human thyroid was identical to that expressed in duodenum with identical sequence deposited in NCBI Genbank of America. (3) Immunohistochemistry, Western blot research and real-time PCR studies showed that motilin and its precursor mRNA were expressed in normal and tumor tissues of human thyroid. Thyroid tumors (acidophilic adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter) showed intense and diffuse immunostaining for motilin peptide. Moreover, the expression of motilin and its precursor mRNA in thyroid medullar carcinoma and acidophilic adenoma were significantly higher than those of normal thyroid tissue (P < 0.05). The expression in thyroid follicular and papillary carcinomas were significantly lower than those of normal thyroid tissue (P < 0.05). There was no difference of the expression between nodular goiter and normal thyroid tissue (P > 0.05).</p><p><b>CONCLUSIONS</b>Motilin peptide and its precursor mRNA are expressed in C cells of human thyroid. The sequence of motilin is identical to that expressed in duodenum from NCBI Genbank of America. The expressions of both motilin and its precursor mRNA in thyroid medullary carcinoma and acidophilic adenoma are significantly increased. In contrast, their expressions in thyroid follicular and papillary carcinomas are significantly decreased. Motilin may regulate physiological functions of the thyroid through parafollicular cells. Motilin may be involved in the pathogenesis of medullary carcinoma and acidophilic adenoma of the thyroid.</p>

Relationship between cortical watershed infarction and carotid artery stenosis and a follow-up and control study on prognosis after stent insertion / 中华神经医学杂志

Objective To investigate the relationship between cortical watershed infarction and carotid artery stenosis and evaluate the stent insertion operation.Methods After 23 cortical watershed infarction patients diagnosed by CT or MRI received DSA detection,we performed stent insertion operationon 11 patients according to their requirements,and conservative treatment on the remaining 12 patients.All the patients underwent follow up for 6-12 months post-operatively.Results Among the 23 cortical watershed infarction patients,22 Were detected with carotid artery stenosis.Statistical analysis showed that the degree of carotid artery stenosis was associated With the elinical svmDtoms and the volume of steal phenomenon(P<0.05);further,the artery stenosis improvement was over 90%with the stent inserted;conversely,dizziness and steal phenomenon disappeared.The post procedure follow-up,ranging 6-12 months,showed that the patients with stent insertion got less new symptoms,steal phenomenon and artery stenosis,compared with the patients with conservation treatment(P<0.05).Conclusion Cortical watershed infarction is associated with carotid artery stenosis.The stent insertion iS useful for the treatment ofcarotid artery stenosis and prevention of cortical watershed infarction.

CT Perfusion Imaging of Ischemic Penumbra in Acute Stroke / 国际脑血管病杂志

Computed tomography perfusion imaging can accurately and promptly provide hemodynamic parameters of brain tissues after stroke,and it has a good correlation with the accepted hemodynamic measurement,the parameters got from xenon-computed tomography, functional magnetic resonance imaging and positron emission tomography.Therefore,CTP has been widely used in predicting the studies of ischemic penumbra after stroke.

Imaging of Perihematoma Penumbra After Intracerebral Hemonrrhage / 国际脑血管病杂志

The prognosis and outcome in patients with intracerebral hemorrhage are closely correlated with perihernatoma tissue injury and recovery.The pathophysiological mechanisms after intracerebral hemorrhage are extremely complicated.There are regional blood flow changes in perihematoma tissues,but whether there is a similar“penumbra”as in cerebral ischemia remains uncertain.At present,studies in this field have mainly focused on the imaging detection of regional cerebral blood flow and metabolism.The study methods frequently used include single photon emission computed tomography,functional magnetic resonance imaging,magnetic resonance spectroscopy,positron emission tomography and CT perfusion imaging,etc.The application of these imaging techniques have provided some valuable methods for identifying whether perihematoma penumbra is present or not,but further studies are needed to confirm it.

Effect of motilin receptor agonist-erythromycin on the glucose responsive neurons in hypothalamus of rats / 中国应用生理学杂志

<p><b>AIM</b>In order to explore the mechanism of central motilin-induced feeding behavior, the effects of erythromycin, a motilin receptor agonist, on glucose responsive neurons in hypothalamus were observed.</p><p><b>METHODS</b>Extracellular recordings were made from single neurons in region of lateral hypothalamic area (LHA) and ventromedial hypothalamic nucleus (VMH) in anesthetized rats. On the basis of their responsiveness to intracarotid injection of 0.58 mol/L glucose solution 0.2 ml, glucose-sensitive neurons (GSNs) in LHA and glucoreceptor neurons (GRNs) in VMH were recognized. Effects of intracerebroventricularly (i. c. v.) administration of 4 microg erythromycin on neural activities of glucose responsive neurons and non-glucose responsive neurons were examined. The mixture of EM and GM-109 1 microl were used to GSNs and GRNs which were sensitive to i. c. v. administration of EM.</p><p><b>RESULTS</b>In LHA, EM increased activity of GSNs significantly (P < 0.05 vs non-glucose-sensitive neurons group). Whereas in VMH, EM significantly decreased the activities of GRNs (P < 0.01 vs non-glucoreceptor neurons group). The mixture of EM and GM-109 had no effect on GSNs and GRNs.</p><p><b>CONCLUSION</b>EM, a motilin receptor agonist, can stimulate GSNs in LHA and suppress GRNs in VMH and this may contribute to central motilin's effect on feeding behavior.</p>