1.Effects of rizatriptan on calcitonin gene-related peptide, proenkephalin and cholecystokinin mRNA expressions in the trigeminal ganglia of a rat migraine model
Gang YAO ; Tingting HAO ; Tingmin YU
Chinese Journal of Neurology 2014;47(9):638-642
Objective This study assesses the influence of rizatriptan on calcitonin gene-related peptide (CGRP),proenkephalin (PENK) and cholecystokinin (CCK) mRNA expressions in the trigeminal ganglia of a rat migraine model and investigates the possible mechanisms by which triptans treat migraine.Methods A total of 24 rats were randomly divided into four groups:normal control group (A),migraine model group(B),rizatriptan control group (C) and rizatriptan treatment group(D).Groups C and D were intragastrically perfused with rizatriptan,1 mg/kg per day.After 7 days,nitroglycerin was subcutaneously injected into the buttocks of the groups B and D to induce migraine.Two hours after nitroglycerin injection,the trigeminal ganglia was isolated.CGRP,PENK and CCK mRNA expressions in the trigeminal ganglia were determined using SYBR Green Ⅰ real-time quantitative PCR.Results The copy number of CGRP mRNA (× 107) in 200 ng total RNA of each group was 0.05 ±0.01,1.30 ±0.52,0.23 ±0.12,0.43 ±0.33 ; The copy number of PENK mRNA (× 103) in 200 ng total RNA of each group was 3.30 ± 1.65,0.34 ±0.14,3.91 ± 2.44,0.71 ± 0.13.The copy number of CGRP mRNA in the trigeminal ganglia of group B was significantly higher than that of group A (q =7.854,P < 0.05) ; CGRP mRNA expressions were significantly lower in the trigeminal ganglia of rats in group D compared with group B (q =5.458,P <0.05).Compared with group A,PENK mRNA expressions in the trigeminal ganglia of rats were significantly lower in group B (q =4.478,P < 0.05).PENK mRNA expressions were significantly higher in trigeminal ganglia of rats in group C compared with group D (q =4.838,P < 0.05).CCK mRNA expression in trigeminal ganglia of rats was similar among groups.Conelusions Rizatriptan can decrease the expressions of CGRP in the trigeminal ganglia of the migraine rats and exhibits neurogenic inflammation triggered by CGRP.PENK expressions decrease in the trigeminal ganglia of the migraine rats,weaken the analgesic effects of enkephalin.
2.Features of chronic daily headache: a clinical analysis of 128 cases
Tingting HAO ; Jingjing QI ; Tingmin YU
Chinese Journal of General Practitioners 2015;14(9):666-669
Objective To review the clinical features of chronic daily headache (CDH).Methods The clinical data of 128 patients with chronic daily headache,including general condition,characteristics of headache,concomitant symptom and disability were analyzed retrospectively.The features of primary chronic daily headache (PCDH) and medication over-dose headache (MOH) were compared.Results Among 128 cases females accounted for 79.7% with an average age of 45.2 years and 88 patients were associated with drug overdose.The symptoms included nausea (68/128),photophobia (75/128),phonophobia (102/ 128),depression (77/128) and irritability (93/128),sleep disorders (94/128),dizziness (75/128),emotional irritability(58/128) and depression(21/128).The migraine disability assessment questionnaire and headache impact test-6 scores showed that disability was resulted from the severe degree of headache in 62.2% (51/82) and 73.2% (82/112) of CDH patients respectively.Compared with PCDH patients,the MOH patients had older age (t =2.59,P =0.011),longer duration (t =2.48,P =0.015) and severer degree of headache(t =5.58,P =0.018),and chronic migraine (t =11.95.P =0.001) was the most common primary headache type.Conclusions Most CDH patients are middle-aged women,with moderate to severe pain,usually complicated with depression,dysphoria and asomnia.Chronic daily headache patients are commonly associated with drug overdose.
3.New progress of anticoagulation therapy in thrombolysis for acute ST-elevation myocardial infarction
Tingting LI ; Yinglu HAO ; Yanping LI
Chinese Journal of Postgraduates of Medicine 2016;(2):182-185
Acute ST-elevation myocardial infarction is a common clinical acute and severe disease, and it has a high fatality rate. At present, thrombolysis is still the most applicable and effective treatment method at the primary hospital. In the thrombolysis therapy, the best anticoagulation therapy can improve the patency of infarct-related coronary artery and lower the rate of reinfarction. For acute ST-elevation myocardial infarction, studies on thrombolysis anticoagulation mechanism, common anticoagulant, bleeding complications and prospect of research are necessary.
4.Expression levels and mechanism of microRNA-34a and Notch1 signaling pathway in hippocampi of temporal lobe epilepsy rats
Baozhen YAO ; Tingting HUANG ; Hao YUAN ; Shiqian YU ; Haijyu ZHANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(12):932-935
Objective To explore the expression levels of microRNA(miR)-34a in hippocampus of temporal lobe epilepsy rats and the effect of miR-34a on its target signaling pathway Notch1.Methods Rats were divided randomly into experiment group (n=40) and control group (n=40) by adopting random number table method.The status epilepticus model and the temporal lobe epilepsy model were induced by using lithium-pilocarpine for experiment group.The control group rats received an injection of an equal amount of 9 g/L saline as instead of pilocarpine.Racine grading was performed at 24 hours,day 3,day 7,day 15,and 1 month after modeling to evaluate the behavior.Real-time fluorescent quantitative polymerase chain reaction was used to test the mRNA expressions of miR-34a and Notch1.Western blot was performed to explore the protein expression of Notch1.Results The expression levels of miR-34a at post-status epilepticus in 24 hours,day 3,day 7,day 15 were 2.55±0.29,2.11±0.17,1.68±0.49 and 1.84±0.42,respectively,which showed statistically significant difference compared with the control group (1.00±0.00) (t=-1.55,-1.11,-0.68,-0.84,all P<0.05).The expression levels of Notch1 mRNA at post-status epilepticus in 24 hours,day 3,day 7,day 15,1 month were 1.44±0.31,1.27±0.13,1.52±0.28,0.91±0.33,and 0.80±0.09 respectively.There were significant differences at 24 hours,day 7 in Notch1 mRNA expression (t=-0.44,-0.52,all P<0.05) compared with the control group(1.00±0.00).The expression level of Notch1 mRNA on day 15 was significantly lower than 24 hours and day 7 (t=-0.54,-0.62,all P<0.05),and the expression in 1 month was significantly lower than in 24 hours,or day 3 and day 7 (t=-0.64,-0.46,-0.72,all P<0.05).The expression levels of Notch1 protein at post-status epilepticus 24 hours,day 3,day 7,day 15,1 month were 0.78±0.09,0.57±0.13,0.55±0.16,0.42±0.13,and 0.33±0.09,respectively.There was significantly up-regulated at 24 hours of Notch1 protein expression compared with control group (0.51±0.15)(t=-0.20,P<0.05);and the expression level at day 15 were significantly lower than 24 hours (t=-0.26,P<0.05),while the expression in 1 month was significantly lower than in 24 hours and on day 3 (t=-0.36,-0.24,all P<0.05).Conclusion miR-34a is significantly up-regulated in the post-status epilepticus rat hippocampus,and it may contribute to temporal lobe epilepsy by activating Notch1 signaling pathway.
5.Osteogenic induction of human bone marrow mesenchymal stem cells cultured in complex medium
Lili XU ; Xiaojuan SUN ; Xiuxian HAO ; Tingting XIE ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2015;(10):1501-1505
BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION:A large number of adherent cels were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cels could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cels. These findings suggest that human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis.
6.Relationship between HLA-DQB1 allele polymorphism with HPV16 infection and cervical cancer susceptibility in Guangxi Zhuang women
Tingting LU ; Huiping LIANG ; Hao XIONG ; Hua ZHU
Chinese Journal of Immunology 2017;33(4):593-597
Objective:To study the relationship of Guangxi Zhuang women being infected by HPV16 and suffering from cervical cancer with HLA-DQB1 allele polymorphism.Provide clues for seeking hereditary susceptibility gene or resistant gene of cervical cancer of Guangxi Zhuang women.Methods:Chose the cervical cancer diagnosed female patients and health women 171 cases respectively aged between 25 and 45 of Guangxi as subject investigated(people in the two groups were paired by age ±3 years).Took their samples to extract HPV DNA and human genome DNA.Then detected HLA-DQB1 alleles and HPV genetype applying PCR-SSP and molecular diversion hybrid technology.Finally the data were statistically analyzed.Results:(1)The total infection rate of HPV in 171 cases of cervical cancer patient was 91.22%,in which the high-risk virus accounted for 90.76%,HPV16 was the main pathogenic subtypes(43.58%).(2)The allele carrying rate of HLA-DQB1*04 in the cervical cancer group was higher than the health control group with statistically significant difference(P<0.05).The allele carrying rate of HLA-DQB1*06/09 in the cervical cancer group was lower than the health control group with statistically significant difference(P<0.05).There was no significant difference of the allele carrying rate of HLA-DQB1*02/05/07/08 between two groups(P>0.05).(3)The occurrence frequency of HLA-DQB1*04 alleles in HPV16 positive cervical cancer patients was significantly higher than HPV16 negative patients with statistically significant difference(P<0.05).Conclusion:HLA-DQB1*04 alleles are probably the susceptibility genes of cervical cancer of Guangxi Zhuang women;HLA-DQB1*06/09 alleles are probably the protective genes of cervical cancer of Guangxi zhuang women;HLA-DQB1*02/05/07/08 alleles seem irrelevant to hereditary susceptibility of cervical cancer of Guangxi Zhuang women.And Guangxi Zhuang women carried HLA-DQB1*04 alleles are more likely to infect HPV16 that increase the risk of cervical cancer.
7.Fragmentation behaviors of taxanes drugs using electrospray ionization with quadrupole time-of-flight mass spectrometry.
Wenxiao MA ; Hao WANG ; Tingting WANG ; Yan YANG ; Jingkai GU
Acta Pharmaceutica Sinica 2013;48(12):1812-6
The fragmentation pathways of two taxanes drugs have been studied in positive ion mode by Q-TOF with the advantages of high mass accuracy and high resolution analysis. The [M+H] + ions were observed by ESI-MS, from which the molecular weights were obtained. Using the protonated pseudo-molecular ions [M+H]+ as internal reference compounds, the accurate mass and element composition of the fragment ions were determined. The collision induced dissociation (CID) data of the [M+H] ions provided fragmentation pathways of related compounds. Results showed that the major cleavage pathways of paclitaxel and docetaxel were the same that the cleavage of C-O bond between the side chain and taxol skeleton easily occurred, then stripping of the functional groups on the parent ring. Some common fragments were formed, such as m/z 105.033 7, 291.137 3, 309.148 5, 327.159 7, 387.181 2 and 509.217 4, which would provide a basis for future qualitative and quantitative analysis of taxanes in vitro and in vivo.
8.Neuroprotective effect of PTEN inhibitor BPV on cerebral ischemia-reperfusion injury in rats and its mechanism
Lunlin MAO ; Donglin HAO ; Xiaowei MAO ; Tingting HUANG ; Bona WU
International Journal of Cerebrovascular Diseases 2015;(9):691-698
Objective To investigated the neuroprotective effect of PTEN inhibitor BPV on cerebral ischemia-reperfusion injury in rats and its mechanism. Methods Male Sprague-Dawley rats were used to induce a reperfusion model of middle cerebral artery occlusion for 1 h. During the reperfusion, the BPV solution (0. 2 mg/kg daily) or the equal volume of saline was injected intraperitonealy immediately. The neurological deficit scores were conducted at day 1, 3,5, and 7 after ischemia-reperfusion. At day 4, triphenyl tetrazolium chloride staining was used to assess cerebral infarction volume. Enzyme-linked immunosorbent assay was used to detect the levels of interleukin 10 (IL-10) and tumor necrosis factor α(TNF-α) in cortical ischemic border zones. Real-time quantitative polymerase chain reaction was used to detect the expression level of PTEN mRNA. Western blotting was used to detect the expression levels of PI3K, Akt, and p-GSK-3β. At day 7, Bielschowsky silver staining was used to detect the axonal distribution in the ischemic border zone of the striatum. Immunohistochemical staining was used to detect the expression of myelin basic protein (MBP). Results At day 4 after ischemia-reperfusion, the infarct volume (32. 27% ± 1. 71% vs. 45. 49% ± 2. 12% ; P < 0. 001), TNF-α concentration in the cortical ischemic border zones (134. 17 ± 10. 38 pg/ml vs. 264. 17 ± 24. 84 pg/ml; P < ), and PTEN mRNA level (1. 19 ± 0. 08 vs. 2. 50 ± 0. 06; P < 0. 001) in the rats of the BPV group were al significantly lower than those of the normal saline group. The IL-10 concentration (186. 83 ± 10. 83 pg/ml vs. 147. 83 ± 11. 62 pg/ml; P < 0. 001), and the expression levels of PI3K (0. 43 ± 0. 08 vs. 0. 26 ± 0. 06; P = 0. 004), Akt (0. 52 ± 0. 05 vs. 0. 40 ± 0. 04;P = 0. 001), and p-GSK-3β (0. 75 ± 0. 08 vs. 0. 38 ± 0. 06; P < 0. 001) were al significantly higher than those of the normal saline group. At day 7 after ischemia-reperfusion, the neurological deficit score (4. 83 ± 0. 41 vs. 6. 33 ± 0. 52; P < 0. 001) in the rats of the BPV group was significantly lower than that of the normal saline group. The axon densities in the ischemic border zones (35. 51% ± 2. 45% vs. 25. 31% ± 2. 79% ; P < 0. 001) and the expression level of MBP (32. 56% ± 3. 46% vs. 27. 81% ± 4. 18% ; P = 0. 037) were significantly higher than those of the normal saline group. Conclusions BPV has neuroprotective effect for cerebral ischemia-reperfusion injury in rats. Its mechanism may be associated with the up-regulation of PTEN downstream proteins PI3K, Akt and p-GSK-3β expression to regulate inflammatory mediators and reduce the inflammatory response.
9.Effects of rizatriptan on behavior and pain-related cytokines in peripheral blood of rat models with nitroglycerin-type migraine
Gang YAO ; Tingting HAO ; Xiangdan LUO ; Tingmin YU
Journal of Jilin University(Medicine Edition) 2014;(5):981-984
Objective To observe the influence of rizatriptan on the behavior and pain-related cytokines in peripheral blood of the migraine model rats, and to investigate the theraputic effect of rizatriptan on migraine.Methods A total of 24 rats were randomly divided into:control group,migraine group,rizatriptan control group and rizatriptan treatment group.The rats in rizatriptan control and rizatriptan treatment groups were intragastrically perfused with rizatriptan,1 mg·kg-1 per day (according to the adult daily dose),and the rats in control and migraine groups were perfused with normal saline,1 mL per day. After 7 d, nitroglycerin was subcutaneously inj ected into the buttocks of the rats in rizatriptan treatment and migraine groups to induce migraine.Normal saline was injected into the rats in control and rizatriptan control groups.At 60-90 min following nitroglycerin injection,the total number of behavioral symptoms was measured.The serum calcitonin gene-related peptide(CGRP),5-hydroxytryptamine (5-HT ), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) levels were determined using ELISA. Results Compared with migraine group, the behavioral score of the rats in rizatriptan treatment group was significantly decreased (P<0.05).The serum CGRP level of the rats in migraine group was significantly higher than that in control group (P<0.05).Compared with control group,the serum 5-HT level of the rats in and migraine group was decreased, but there was no significant differece (P>0.05 );the serum 5-HT level in rizatriptan control group was significantly increased(P<0.05).The serum 5-HT level of the rats in rizatriptan treatment group was significantly increased compared with migraine group (P<0.05).The serum IL-1βand TNF-αlevels of the rats in varions groups had no significant differences (P>0.05).Conclusion Rizatriptan can relieve the behavior symptoms in nitroglycerin-induced migraine model rats, increase the serum 5-HT level, improve the vasomotor disturbance,and relieve the angiectasis.
10.Rizatriptan effects on the cholecystokinin expression in endogenous pain modulatory system of a rat mi-graine model
Gang YAO ; Tingting HAO ; Qian HUANG ; Tingmin YU
Chinese Journal of Behavioral Medicine and Brain Science 2014;(11):961-963
Objective To assess the influence of Rizatriptan on the cholecystokinin( CCK) expression in periaqueductal gray( PAG) of migraine model rat to investigate the possible mechanism by which Triptans treat mi?graine. Methods A total of 24 rats were randomly divided into four groups:normal control groups(A),migraine model groups(B),Rizatriptan control groups(C) and Rizatriptan treatment groups(D).C and D groups were intra?gastrically perfused with Rizatriptan,1 mg/kg per day. After 7 days,nitroglycerin was subcutaneously injected into the buttocks of the B and D group to induce migraine. Two hours after nitroglycerin injection,the trigeminal ganglia were isolated.CGRP expression in periaqueductal gray were determined using SYBR Green I real?time quantitative PCR and Immunohistochemistry. Results CCK mRNA levels ( target gene mRNA copies per 250 ng total RNA,× 106) in the rat midbrain of A,B,C,D groups were 1.25±0.41,1.71±0.93,0.17±0.12,0.22±0.07 respectively. CCK?8?immunoreactive positive cells in the rat PAG of each group were 37.17±12.62,40.17±11.09,27.33±7.71, 20.67±7.66 respectively. CCK mRNA expression in group C was significantly lower than that of group A(P<0.05) while the CCK mRNA expression in group D was lower than that of group B(P<0.05).The CCK?8?positive cells of the rat PAG in group D were lower than that in group B(P<0.05) . Conclusion Rizatriptan can down regulate the expression of CCK?8 in the PAG of the migraine rats and weaken the CCK?8 induced inhibition of the analgesic effects of opioid peptides.