2.Changes on degranulation of mast cells and neurogenic inflammation-related factors in the dura mater of the rat model of migraine
Wu XU ; Zhaochun SHI ; Junchao WEI ; Yuezhou CAO ; Ting WU ; Qi WAN
Chinese Journal of Neurology 2011;44(8):563-567
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.
3.Measurement of mass fraction of Sanjie Zhentong capsules with near infrared reflectance spectroscopy.
Guang-ren ZHANG ; Yun WU ; Rui-ting JIN ; Qin WAN ; Jin-feng WU ; Xiao-qian XU ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(16):3204-3208
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
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chemistry
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Drugs, Chinese Herbal
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chemistry
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Least-Squares Analysis
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Spectroscopy, Near-Infrared
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methods
4.Optimization of short tandem repeats and their application in prenatal diagnosis of spinal muscular atrophy
Jun-Fen SU ; Wan-Jin CHEN ; Zhi-Ying WU ; Ning WANG ; Yu LIN ; Min-Ting LIN ; Shenxing MURONG ;
Chinese Journal of Neurology 2005;0(07):-
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.
5.Preparation of polyclonal antibody against survival motor neuron protein and study on the expression of survival motor neuron protein in the skeletal muscular of patients with spinal muscular atrophy
Wan-Fin CHEN ; Zhi-Ying WU ; Ning WANG ; Jun-Feng SU ; Min-Ting LIN ; Shen-Xing MURONG ;
Chinese Journal of Neurology 2005;0(12):-
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.
6.Analysis of the Efficacy and Prognosis Factors of Acute Myeloid Leukemia with a Combination Therapy of Venetoclax
Chong-Chong REN ; Wan-Wan ZHANG ; Ting-Kai WU ; Bei LIU
Journal of Experimental Hematology 2024;32(1):104-111
Objective:To explore the efficacy and prognosis factors of acute myeloid leukemia with a combination therapy of venetoclax.Methods:A retrospective analysis was performed on the clinical data of AML patients treated with a combination therapy of venetoclax from March 2020 to April 2023 in the First Hospital of Lanzhou University.The efficacy,adverse reactions and survival were observed,and the influencing factors were analyzed.Results:A total of 74 AML patients were included in this study,including 43 initially treated AML and 31 relapsed or refractory AML(R/R AML).The median age of 43 initially treated AML patients was 65 years old,the composite complete remission(cCR)rate was 67.4%(29/43),the objective response rate(ORR)was 72.1%(31/43),and the median overall survival(OS)was 17.3 months.The median age of 31 R/R AML patients was 51 years old,with a cCR rate of 38.7%(12/31),an ORR of 58.1%(18/31),and a median OS of 7.1 months.Sex,the blood cell count before VEN,gene mutation and prognosis stratification were related to whether to obtain cCR.Failure to obtain cCR was an independent risk factor for adverse outcomes.Conclusion:A combination therapy of venetoclax is safe and efficacious for AML.Its efficacy and survival are affected by molecular biology,cytogenetics and other factors.
7.The Correlation of Gene Mutation and Clinical Characteristics in Patients with Myelodysplastic Syndrome and Prognostic Analysis
Wan-Wan ZHANG ; Ya-Li ZHANG ; Chong-Chong REN ; Ting-Kai WU ; Bei LIU
Journal of Experimental Hematology 2024;32(1):176-183
Objective:To explore the correlation between gene mutations and clinical characteristics,prognosis of myelodysplastic syndromes(MDS).Methods:Clinical data of 131 patients with MDS were collected from the First Hospital of Lanzhou University from June 2015 to February 2023,which 19 of them developed into secondary acute myeloid leukemia(sAML)during follow-up time.Second generation sequencing technology was used to detect the mutation types of MDS disease-related genes,drawn gene maps,and analyzed their correlation and prognosis based on the clinical data of patients.Results:The median age of 131 MDS patients was 58(17-86)years old.The ratio of male to female was 1.3:1.A total of 148 gene mutations and 25 types were found in the center.U2AF1 and ASXL1 were often co-mutations with other genes,which were accompanied by 20q-and normal karyotype(NK)respectively.SETBP1 and SRSF2 were more common in patients over 60 years old,while NPM1 and WT1 under 60 years.Older patients had a higher the number of genetic mutations than younger patients.The incidence of SF3B1 and RUNX1 in males was higher than females and DNMT3A in females was higher than males.The number of gene mutations in sAML was higher than MDS(1.8 vs 1.0,P=0.006).The univariate and multivariate analysis showed that IPSS-R prognostic score≥ 3.5,TP53 were adverse factors for poor prognosis in MDS patients.Patients with monoallelic mutation(ma-TP53)and wild-type(wt-TP53)TP53 had OS better than biallelic mutation(bi-TP53)(P=0.003).The OS of MDS patients was better than sAML(P=0.01)and transplant patients was significantly better than non-transplant patients(P=0.036).Conclusion:Gene mutation is closely related to cytogenetic indexes and clinical features(peripheral blood cell count,sex,age).IPSS-R prognostic score and TP53 were risk factors affecting OS in MDS patients.
8.Phenotypic transformation of corpus cavernosum smooth muscle cells in hypertensive rats.
An-yang WEI ; Wei-wu WU ; Hao-bin WU ; Ting-yu YE ; Bo WAN
Journal of Southern Medical University 2009;29(5):925-928
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
9.Expression of human telomerase reverse transcriptase and c-myc transcripts in endometrial carcinoma and its precursors.
Ying DONG ; Ting LI ; Ying WANG ; Hai-rong WU ; Min XIE ; Wan-zhong ZOU
Chinese Journal of Pathology 2004;33(1):40-43
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
10.The Associations between Visual Attention and Facial Expression Identification in Patients with Schizophrenia.
I Mei LIN ; Sheng Yu FAN ; Tiao Lai HUANG ; Wan Ting WU ; Shi Ming LI
Psychiatry Investigation 2013;10(4):393-398
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
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Automatic Data Processing
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Cognition
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Facial Expression*
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Humans
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Schizophrenia*