Arteriovenous Fistula ; complications ; etiology ; Cardiac Output, High ; Femoral Artery ; Femoral Vein ; Heart Failure ; etiology ; Humans ; Intra-Aortic Balloon Pumping ; adverse effects ; Male ; Middle Aged

Objective To observe the changes on the neurogenic inflammation-related factors in the dura mater of the rat model of migraine and investigate the possible mechanism of the pain of migraine.Methods SD rats were randomly divided into stimulation group ( n = 32 ) and sham group ( n = 32 ).Unilateral trigeminal ganglion was stimulated to induce migraine for rats in the stimulation group. Rats in the sham group were subjected to sham surgery. The levels of calcitonin gene-related peptide (CGRP) in the blood of jugular vein in the stimulation side were measured by radioimmunoassay. The levels of histamine in peripheral blood and prostaglandin E2 (PGE2 ) in the dura mater were determined by enzyme-linked immunosorbent assay (ELISA). The number of mast cells and percentage of their degranulation in the dura mater were determined under a microscope after toluidine blue staining. Cyclooxygenase 2 (COX-2)expression in the dura mater was evaluated by immunohistochemical staining and western blot analysis. Results In the stimulation group, the level of CGRP in the ipsilateral jugular vein was (82. 84 ± 16. 24)pg/ml and in the sham group was (59. 20 ±11.66) pg/ml (t = -3.34, P ＜ 0. 05 ). The level of histamine in the ipsilateral jugular vein was ( 11.59 ± 1.20) ng/ml and in the sham group was (9. 87 ±0. 88) ng/ml (t = - 3. 27, P ＜ 0. 05). The number of mast cells in the dura mater decreased from 15.46 ± 2. 40 in the stimulation group to 11.63 ± 1.67 in the sham group ( t = 3.71, P ＜ 0. 05 ). Degranulation of mast cells in the dura mater significantly increased from 14. 09% ±4. 53% in the sham group to 29. 10% ±9. 39% in the stimulation group (t = - 4. 07, P ＜ 0. 05 ). The level of PGE2 in the stimulation group was ( 382. 30 ±20. 90) pg/ml and in the sham group was (80. 70 ± 10. 60) pg/ml (t = - 16. 674, P ＜0. 05). The number of COX-2 positive cells significantly increased from 42. 00 ± 18.40 in the sham group to 139.00 ±20. 50 in the stimulation group (t = -7. 994, P ＜0. 05). Also the COX-2 protein level was elevated from 19. 50 ±9. 20 in the sham group to 359. 20 ±21.90 in the stimulation group (t = -5. 190, P ＜0. 05). Conclusions Electrical stimulation on the unilateral trigeminal ganglion induces neurogenic inflammation in the dura mater. Changes on the neurogenic inflammation-related factors are probably the essential pathophysiological mechanism underlying the pain in migraine.

Sanjie Zhentong capsules were scanned by using a near infrared spectra probe with different drug mass fraction and the spectral information of capsule shells and contents in it were obtained. Then partial least squares (PLS) models were developed for the prediction of mass fraction of Fritillariae Thunbergii Bulbus and Resine draconis in Sanjie Zhentong capsules. The correlation coefficient (r9c)) and root mean standard error( RMSEC) of 0.949 5, 0.958 2 and 4.742 4, 4.135 7. The models obtained correlation coefficient (r(v)) of 0.919 2, 0.936 7 and root mean square error (RMSECV) of 6.158 9, 5.037 3 respectively in the training set. The paired T test analysis of statistics showed that there were no significant difference between predictive values and measure values. The established models reflected a strong prediction performance and can meet the needs of the production.
Capsules ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Spectroscopy, Near-Infrared ; methods

Objective To optimize the short tandem repeats(STR)which link closely to survival motor neuron(SMN)and have redundant polymorphism information contents,and to use these STR in the prenatal diagnosis of spinal muscular atrophy(SMA).Methods Eleven STR loci(D5S435,D5F153, DSF151,D5S637,D5S1413,D5S125,D5S464,D5S1556,DSF149,D5S351,MAP1B-5')were amplified by PCR.Then the PCR products were detected by polyacrylamide gel electrophoresis(PAGE)and analyzed by silver staining.STR loci were evaluated and optimized by their PIC values.PCR-PAGE and gene scan were combined to make genetic link analysis for SMA families based on the optimized STR.Results Three STR loci(D5S435,DSF149 and D5S351)were selected with 8,19 and 18 polymorphic fragments detected respectively in 100 normal individuals.Their PIC values were 0.84,0.91 and 0.92 respectively.Four carriers and 2 normal individuals were detected from 6 SMA families with linkage analysis by using the 3 STR.Conclusion This genetic diagnosis system based on the 3 STR loci can provide rapid prenatal diagnosis for SMA families,can eliminate maternal blood contamination,and also can discriminate carriers from normal individuals in the fetuses,which makes the prenatal diagnosis system of SMA perfect.

Objective To prepare the survival motor neuron(SMN)polyclonal antibody and explore the localization of SMN protein in transfected cells and its expression in skeletal muscles of patients with spinal muscular atrophy(SMA).Methods A prokaryotic expressional plasmid named pET-28? (+)/SMN was constructed and SMN-His fusion protein was induced.The fusion protein was used to immunize New Zealadd rabbits to prepare SMN polyclonal antibody.A eukaryotic expressional plasmid named pcDNA3.1/myc-HisB-SMN was constructed and used to transfect CHO cells.Skeletal muscles were collected from 3 patients with bone fracture who were regarded as normal controls, and 3 SMA patients of type Ⅰ, 3 of type Ⅱ and 3 of type Ⅲ who were ascertained by genetic analysis.Western-blotting and immunofluorescence stain were applied to study the expression of SMN in transfected CHO cells and skeletal muscles of normal individuals and SMA patients.Results Correct pET-28a(+)/SMN prokaryotic expressive plasmid was constructed and SMN-His fusion protein was obtained from E coli BL21 transformed with pET-28a(+)/SMN.Then, rabbit anti-human full-length SMN polyclonal antibody of high specificity and sensitivity was obtained from rabbits immunized by SMN-His fusion protein.SMN proteins were shown diffusedly locating in the cytoplasm and nucleus of CHO cells transfected with pcDNA3.1/myc-HisB-SMN plasmid and mainly accumulating around the nucleus.The results of Western-blotting were as follows:the average ratio of SMN band density to glyceraldehyde phosphate dehydrogenase(GAPDH)band density (SMN/GAPDH)is 0.619 in skeletal muscles from normal controls, the average values of SMN/GAPDH in skeletal muscle from SMA patients of type Ⅲ and Ⅱ were 0.347 and 0.340 respectively, which were lower than that of normal controls.However, the average values of SMN/GAPDH in skeletal muscle from SMA patients of type I was only 0.079, which was quite lower than that of normal controls.Conclusions The rabbit anti-human full-length SMN polyclonal antibody is of high specificity and sensitivity, which makes the basis for the research of SMN function and SMA pathogenesis.There may be a correlation between the SMN level in skeletal muscle and the severity of disease.

OBJECTIVETo validate the hypothesis that the phenotypic transformation occurs in the smooth muscle cells in the corpus cavernosum of hypertensive rat and explore its impact on the erectile function of rats.

METHODSEighteen 16-week-old male spontaneously hypertensive rats (SHR) and 10 syngeneic normotensive rats (WKY) were used in this experiment. After measurement of systolic blood pressure of the caudal artery and examination of the erectile function with subcutaneous injection of apomorphine (APO), the rats were divided into 3 groups, namely hypertensive with erectile dysfunction (HBP-ED) group (n=6), hypertensive (HBP) group (n=12) and control group (n=10). Immunohistochemical staining and color image analysis system were used to observe expression of calponin 1 and osteopontin (OPN) in rat corpus cavernosum. Real-time quantitative RT-PCR was used to determine the expression of calponin 1 and OPN mRNAs in different groups.

RESULTSThe expressions of calponin 1 protein and mRNA were the highest in the control group and the lowest in HBP-ED group, while the expressions of OPN protein and mRNA were the highest in HBP-ED group and the lowest in the control group.

CONCLUSIONThe smooth muscle cells may transform from the contractile phenotype into synthetic phenotype in the corpus cavernosum of the hypertensive rats, resulting ultimately in erectile dysfunction.

Animals ; Calcium-Binding Proteins ; genetics ; metabolism ; Erectile Dysfunction ; etiology ; pathology ; Hypertension ; complications ; pathology ; Male ; Microfilament Proteins ; genetics ; metabolism ; Myocytes, Smooth Muscle ; pathology ; Osteopontin ; genetics ; metabolism ; Penis ; pathology ; Phenotype ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR

OBJECTIVETo study the possible role of hTERT and c-myc in endometrial carcinogenesis.

METHODSThe expression of hTERT and c-myc mRNA was examined by in situ hybridization of endometrial samples from 14 cases with simple hyperplasia, 10 with complex hyperplasia, 8 with atypical hyperplasia and 42 with endometrioid carcinoma.

RESULTSExpression of hTERT was demonstrated in samples with simple hyperplasia, complex hyperplasia, atypical hyperplasia and carcinoma at frequencies of 2/14, 4/8, 8/10 and 39/42 (92.9%), respectively. The prevalence and intensity of the hTERT signal was greater in the carcinomas and lesions with atypical hyperplasia than those with simple or complex hyperplasia (P < 0.05). The expression of c-myc was demonstrated in samples with simple hyperplasia, complex hyperplasia, atypical hyperplasia and carcinoma at frequencies of 3/14, 1/8, 5/10 and 23/42 (54.8%), respectively. The frequency of c-myc expression was higher in carcinomas and hyperplastic lesions with atypia than those in lesions with simple or complex hyperplasia without atypia (P < 0.05). The expression of hTERT was shown to be correlated with the level of differentiation (P < 0.05), while the c-myc expression appeared to be associated with the depth of myometrial invasion (P < 0.05). The expression levels of hTERT and c-myc were not found to be correlated with each other in the tissues examined (P > 0.05).

CONCLUSIONSThe expression of hTERT and c-myc may be involved in the progression from the endometrial aypical hyperplasia to invasive carcinoma. The correlation between hTERT and c-myc in endometrial hyperplasia and carcinoma are not found.

Adult ; Aged ; DNA-Binding Proteins ; Endometrial Neoplasms ; genetics ; pathology ; Female ; Genes, myc ; Humans ; Male ; Middle Aged ; Precancerous Conditions ; genetics ; pathology ; RNA, Messenger ; analysis ; Telomerase ; genetics

OBJECTIVETo construct a method for detecting the copy number of survival of motor neuron 1 gene (SMN1) with single copy difference based on real-time fluorescence quantitative PCR, and to make practical use of the method for acquiring the data on SMN1 copy number in Chinese as well as for screening the carriers of spinal muscular atrophy (SMA) from healthy individuals and SMA families.

METHODSExon 7 and flanking area of SMN1 gene were amplified by real-time fluorescence quantitative PCR in 264 healthy individuals, in 1 standard sample having 2 SMN1 but having no SMN2, and in 88 parents of SMA patients. The samples for detecting were diluted to 30 ng/microL and the standard sample was diluted to 15 ng/microL, 30 ng/microL, 45 ng/microL, 60 ng/microL; the unknown samples and 4 standard samples with different concentrations were amplified at the same time, a standard curve could be drawn out according to the results of the 4 standard samples, then the copy number of samples could be calculated.

RESULTSOf 88 parents' samples, 84 samples each had 1 copy of SMN1, and the rest 4 each had 2 copies of SMN1. Of 264 healthy individuals' samples, 5 samples each had only 1 copy of SMN1 (an indicator of definite gene carriers), 232 samples each had 2 copies of SMN1, 25 samples each had 3 copies of SMN1, and 2 samples each had 4 copies of SMN1. Of the samples of 32 members of SMA families, 2 samples each had only 1 copy of SMN1 indicating definite gene carriers, 25 samples each had 2 copies of SMN1, and 5 samples each had 3 copies of SMN1.

CONCLUSIONSMN1 copy number could be detected precisely by real-time fluorescence quantitative PCR; the screening of gene carriers could provide essential data for genetic counseling.

Exons ; Family Health ; Female ; Fluorescence ; Gene Dosage ; Humans ; Male ; Muscular Atrophy, Spinal ; genetics ; Polymerase Chain Reaction ; methods ; Survival of Motor Neuron 1 Protein ; genetics

OBJECTIVE: Visual search is an important attention process that precedes the information processing. Visual search also mediates the relationship between cognition function (attention) and social cognition (such as facial expression identification). However, the association between visual attention and social cognition in patients with schizophrenia remains unknown. The purposes of this study were to examine the differences in visual search performance and facial expression identification between patients with schizophrenia and normal controls, and to explore the relationship between visual search performance and facial expression identification in patients with schizophrenia. METHODS: Fourteen patients with schizophrenia (mean age=46.36+/-6.74) and 15 normal controls (mean age=40.87+/-9.33) participated this study. The visual search task, including feature search and conjunction search, and Japanese and Caucasian Facial Expression of Emotion were administered. RESULTS: Patients with schizophrenia had worse visual search performance both in feature search and conjunction search than normal controls, as well as had worse facial expression identification, especially in surprised and sadness. In addition, there were negative associations between visual search performance and facial expression identification in patients with schizophrenia, especially in surprised and sadness. However, this phenomenon was not showed in normal controls. CONCLUSION: Patients with schizophrenia who had visual search deficits had the impairment on facial expression identification. Increasing ability of visual search and facial expression identification may improve their social function and interpersonal relationship.
Asian Continental Ancestry Group ; Automatic Data Processing ; Cognition ; Facial Expression* ; Humans ; Schizophrenia*

OBJECTIVETo introduce the application of denaturing high-performance liquid chromatography (DHPLC) in the diagnosis of childhood type spinal muscular atrophy (SMA).

METHODSExon 7 and flanking area of survival motor neuron (SMN) gene were amplified by PCR in 1 standard sample, 25 normal individuals and 25 patients with SMA. The PCR products were then directly loaded onto the DHPLC system after denaturing and annealing. Different DNA segments were separated by changing the concentration of buffer A relative to that of buffer B.

RESULTSDifferent DNA segments were separable on the DHPLC chromatogram. Three peaks including SMN1/SMN2 heteroduplex peak, SMN2 homoduplex peak and SMN1 homoduplex peak were detected in 23 out of 25 normal individuals. Only SMN1 homoduplex peak was detected in 2 normal individuals and the standard sample, indicating the deletion of SMN2 On the contrary, only the SMN2 homoduplex peak was detected in 22 out of 25 patients with SMA, indicating deletion of SMN1. The three peaks as those of normal individuals were detected in the other 3 patients, indicating no SMN1 or SMN2 deletion.

CONCLUSIONAs a new technology for diagnosing SMA, DHPLC is sensitive, accurate, rapid and convenient.

Chromatography, High Pressure Liquid ; methods ; Exons ; genetics ; Humans ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; SMN Complex Proteins ; genetics ; Sensitivity and Specificity ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein