1.Construction of pRluc-hNTSR1-pcDNA3.1 eukaryotic expression vectors and its expression in isolated cells
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(12):1140-1142
Objective To construct expression vectors that Renilla reniformis (Rluc) fused with neurotensin type 1 receptor (NTSR1),and to investigate the interaction between NTSR1 and other receptors,as well as intercellular signal transduction mechanism mediated by neurotensinl-R.Methods The human NTSR1 gene was amplified by PCR using the plasmid pcDNA3.1-hNTSR1 as template.The PCR product was digested,ligased with the plasmid pRluc and then be transformed into the competent cell Top10.The construct was identified by DNA sequencing.The recombinant plasmid was transiently transfected into human embryonic kidney 293 ( HEK293 )cells,and the expression of pRluc-hNTSR1-pcDNA3.1 was detected by confocal microscopy and Western blot.Results The fragment of 1257 bp was amplified by PCR,and the DNA sequences were identical with the gene in GenBank ( NM_002531 ).Western blot showed a band about 90kDa.Confocal microscopy showed that NTSR1 was expressed on the plasma membrane.Conclusion The pRluc-hNTSR1-pcDNA3.1 eukaryotic expression vector is successfully constructed,and the expression vector can be used to investigate the interaction between NTSR1 and other receptors,as well as intercellular signal transduction mechanism mediated by neurotensinl-R,which will provide new target for drug development.
2.Role of G-protein-coupled receptor kinases in cell migration and signal transduction
Xin CAI ; Jing CHEN ; Jingmei ZHANG ; Bo BAI
Chinese Journal of Pathophysiology 2011;27(7):1438-1444
G-protein-coupled receptor kinases (GRKs) are a family of serine/threonine protein kinases. The investigators pay much attention to the roles of GRKs in the signal transduction through G-protein-coupled receptors (GPCRs) with arrestin ever since a long time ago. Due to the physiological and pathological observations with the methods of deletion or overexpression, GRKs are considered as new drug targets. The kinases play a role in the pathogenesis of hypertension and cell migration through GPCRs and Hedgehog signaling pathways. As the development of research techniques, especially bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), the special mechanism of GRKs for GPCRs is more evident. In this review, we discuss the recent achievement in the roles of GRKs signaling and the related newest research techniques.
3.Laborotary study of DanHong in improving the movement function after stroke
Wei WANG ; Guangran CUI ; Tianyi MA ; Juanjuan XIN ; Bo LIU ; Zhiping CAI
Journal of Regional Anatomy and Operative Surgery 2015;(2):157-159
Objective To investigate Danhong in improving the movement function of reperfused rat after stroke. Methods Rats to make middle cerebral artery occlusion model,after 24 hours of reperfusion were divided into control group,high dose and low dose of Danhong groups and Venorruton group randomly. Rats in Control group were injected saline. High and low dose group were injected DanHong according to their weight,high dose as 14. 4 mg/kg,low dose as 3. 6 mg/kg. Venorruton group were injected Venorruton as 0. 04 g/kg. The infarcted square of rat brain was measured,the rats were tested with walking wood bar and two fore limb grasp strength. Results Rat infarcted squre in 24 hours and 3 days,control groups is the biggest,compared with others,there are statistically difference(P<0. 05). Score in each group of walking wood bar in 3rd days are (1. 30 ± 0. 91),(3. 78 ± 1. 72),(4. 18 ± 2. 05),(4. 63 ± 2. 45). Compared between control and other druge groups,there are statistically differnec(P<0. 05). There are statistically differences between control and other groups in two fore limb grasp testing(P<0. 05). Conclusion Danhong has the same effect which can improve the movement function to stroke rat as Venorruton.
4.Prevalence of fatty liver in a district of Shanghai detected by B-type ultrasonography and serum activity of alanine aminotransferase
Xiao-Bo CAI ; Jian-Gao FAN ; Jun ZHU ; Xin-Jian LI ; Rui LI ; Fei DAI ;
Chinese Journal of General Practitioners 2005;0(10):-
Objective To study the prevalence of fatty liver and its risk factors in adult population of Pudong New District,Shanghai detected by combination of B-type uhrasonographic features and elevated serum activity of alanine aminotransferase (ALT).Methods A cross-sectional survey was performed in 2017 residents aged 16 years over recruited from four neighborhoods of Prdong New District of Shanghai with multi-phase cluster sampling,including interview with questionnaire,physical check-up,anthropometry, measurement of plasma glucose and lipid profile,ALT activity and real-time B-type ultrasnnography.Serum hepatitis B surface antigen (HBsAg) was further detected for those with elevated ALT activity.Results Prevalence of fatty liver was 21.32 percent (430/2017) in the residents of the District participated in this survey.Prevalence of abdominal obesity,hypertriglyceridemia,hyperlipoproteinemia (low-density lipoprotein-cholecterol),essential hypertension,impaired glucose tolerance,diabetes and metabolic syndrome were 71.16,71.16,11.86,66.74,35.58,24.40 and 47.21 percent in those with fatty liver, respectively,as compared to 26.34,12.73,4.79,39.57,24.01,6.81 and 11.28 percent in those without fatty liver (controls),respectively.Multivariate logistic regression analysis showed that body mass index, 2-h postprandial glucose level,diastolic blood pressure,serum level of triglyceride,abdominal obesity and diabetes all were independent risk factor for tatty liver,with odds ratio (OR) of 1.080,1.149,1.035, 1.526,1.960 and 1.391,respectively.Conclusions Prevalence of fatty liver was relatively high in Shanghai Pudong New District.Fatty liver closely associates with disturbance of carbohydrate and lipid metabolism.
5.Expression profile of a novel germ cell-specific gene, TSCPA, in mice and human.
Zhendong, YU ; Bo, WU ; Aifa, TANG ; Jing, CHEN ; Xin, GUO ; Jie, QIN ; Yaoting, GUI ; Zhiming, CAI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):535-9
In order to identify novel genes involved in spermatogenesis, testis cDNA samples from Balb/C mice of different postnatal days were hybridized with the whole mouse genome Affymetrix chip to screen the testis-specific genes. The characteristics of the selected genes were analyzed by RT-PCR as well as other bioinformatic tools. A novel differentially expressed testis-specific gene (GenBank Accession No: NM_029042) in the developmental stages of testes was identified, and named TSCPA. Cellular mapping prediction of TSCPA indicated that its protein was probably expressed in nuclei, and one putative domain (aa 332-377) was anchoring domain of cAMP-dependent type II PK. The result of subcellular localization of GFP-TSCPA fusion protein in Cos-7 cells showed that TSCPA protein was expressed in nuclei. RT-PCR analysis revealed that TSCPA was expressed specifically in mouse and human testis. TSCPA gene was expressed weakly in 21-day-old mouse testis and the expression was increased gradually from 38th day to 6th month of mouse testes. No expression of hTSCPA was found in cryptorchidism and Sertoli-cell-only syndrome patients. It was concluded that the expression profile of TSCPA in human and mice indicated that TSCPA might play an important role in spermatogenesis.
6.Expression of Aquaporin 4 in Diffuse Brain Injury of Rats.
Ren-hui CHEN ; Song-guo HE ; Can-xin CAI ; Bo-xue HUANG ; Zhi-rong WANG
Journal of Forensic Medicine 2016;32(1):18-25
OBJECTIVE:
To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema.
METHODS:
The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4.
RESULTS:
Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased.
CONCLUSION
The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.
Animals
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Aquaporin 4/metabolism*
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Astrocytes
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Blood-Brain Barrier/metabolism*
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Brain
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Brain Edema/metabolism*
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Brain Injuries/metabolism*
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Cell Membrane Permeability/genetics*
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Disease Models, Animal
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Permeability
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Rats
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Water
7.Determination of the interaction kinetics between meloxicam and β-cyclodextrin using the quantitative high-performance affinity chromatography coupled with mass spectrometry.
Cai-fen WANG ; Zhuo LI ; Xiao-bo WANG ; Hai-yan LI ; Ji-wen ZHANG ; Li-xin SUN
Acta Pharmaceutica Sinica 2015;50(9):1167-1173
The association rate constant and dissociation rate constant are important parameters of the drug-cyclodextrin supermolecule systems, which determine the dissociation of drugs from the complex and the further in vivo absorption of drugs. However, the current studies of drug-cyclodextrin interactions mostly focus on the thermodynamic parameter of equilibrium constants (K). In this paper, a method based on quantitative high performance affinity chromatography coupled with mass spectrometry was developed to determine the apparent dissociation rate constant (k(off,app)) of drug-cyclodextrin supermolecule systems. This method was employed to measure the k(off,app) of meloxicam and acetaminophen. Firstly, chromatographic peaks of drugs and non-retained solute (uracil) on β-cyclodextrin column at different flow rates were acquired, and the retention time and variance values were obtained via the fitting the peaks. Then, the plate heights of drugs (H(R)) and uracil (H(M,C)) were calculated. The plate height of theoretical non-retained solute (H(M,T)) was calculated based on the differences of diffusion coefficient and the stagnant mobile phase mass transfer between drugs and uracil. Finally, the k(off,app) was calculated from the slope of the regression equation between (H(R)-H(M,T)) and uk/(1+k)2, (0.13 ± 0.00) s(-1) and (4.83 ± 0.10) s(-1) for meloxicam and acetaminophen (control drug), respectively. In addition, the apparent association rate constant (k(on,app)) was also calculated through the product of K (12.53 L x mol(-1)) and k(off,app). In summary, it has been proved that the method established in our study was simple, efficiently fast and reproducible for investigation on the kinetics of drug-cyclodextrin interactions.
Acetaminophen
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chemistry
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Chromatography, Affinity
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Drug Interactions
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Kinetics
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Mass Spectrometry
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Thermodynamics
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Thiazines
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chemistry
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Thiazoles
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chemistry
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beta-Cyclodextrins
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chemistry
8.Finite element analysis of acetabular medial wall displacement osteotomy in adult acetabular dysplasia
Dongsong LI ; Shuqiang LI ; Bo CAI ; Zhengang ZHAO ; Jikui GUAN ; Chen YANG ; Wei FENG ; Xin QI ; Jianguo LIU
Chinese Journal of Tissue Engineering Research 2010;14(48):9104-9108
BACKGROUND: Adult acetabular dysplasia in advanced stage combined with hip joint osteoarthritis should undergo total hip replacement. The severity of acetabular lesion is various in different patients, which leads to significantly increased difficulty in reestablishing acetabulum. Acetabular medial wall displacement osteotomy can solve the component of acetabular prosthesis, but the displacement range of the acetabular medial wall following osteotomy is controversial.OBJECTIVE: To look for a suitable displacement range of acetabular medial wall following osteotomy by computer-aided design finite element analysis.METHODS: SolidWorks 2008 software was used to establish three-dimensional models of acetabular dysplasia pelvis. Acetabular medial wall displacement osteotomy was simulated to make acetabular medial wall bone displace from 2 mm bone contact to 7 mm bone contact in the pelvic cavity. One experimental group was set at 1 mm intervals, totally 10 experimental groups. The acetabulum in each group was split into four quadrants. The prosthesis acetabulum-bone interface in each group was analyzed by computer simulation contrast mechanics experiment. The Mises stress and shear stress values were measured between acetabular prosthesis and bone interface.RESULTS AND CONCLUSION: In groups 1, 5, 6, 9 and 10, the Mises stress was unevenly distributed in posterior inferior, anterior superior and anterior inferior quadrants. In groups 2, 3, 4, 7 and 8, the Mises stress was evenly distributed in posterior inferior,anterior superior and anterior inferior quadrants. Of them, the stress was most even in the group 4. In groups 2, 3, 4, 7 and 8, the shear stress was evenly distributed in the above-mentioned three quadrants. The shear stress was lowest in the groups 7 and 8.These indicate that joint force in the acetabulum mainly focused in the posterior superior quadrant. With the displacement of acetabular cup, the contact area of acetabular cup and bone would gradually increase, which finally increased the Mises stress in the contact surface. However, shear stress decreased with displacement of acetabular medial wall. Therefore, the suitable displacement range of acetabular medial wall osteotomy is 1 mm away from the pelvic cavity and 1 mm complete embolism in the pelvic cavity. The optimal position was 1 mm complete embolism in the pelvic cavity.
9.Therapeutic Efficacy of Goserelin Acetate Sustained-release Implants Combined with Bicalutamide in the Treatment of Elderly Prostate Cancer Patients and Cognitive Function and Short-term Survival Rate Study
China Pharmacy 2018;29(3):382-385
OBJECTIVE: To investigate therapeutic efficacy of Goserelin acetate sustained-release implants combined with bicalutamide in the treatment of elderly (≥70 years old) prostate cancer patients, and its effects on cognitive function and short-term survival rate. METHODS: A total of 56 prostate cancer patients treated in our hospital from Nov. 2014 to Nov. 2016 were divided into observation group and control group according to random number table, with 28 cases in each group. Observation group was given maximal androgen blokage (MAB) treatment which was Goserelin acetate sustained-release implant (subcutaneous injection of abdominal wall, 3. 6 mg/ times, once) combined with Bicalutamide tablet (orally, 50 mg/times, qd). Control group received surgical castration, and then was given docetaxel (intravenous dripping on 1st day) combined with Prednisone acetate tablets (lst-21st day, orally, 5 mg/time, bid) after surgery for adjuvant therapy. Treatment course of 2 groups lasted for 3 weeks, and all patients were followed up for 12 months. Clinical efficacy, Montreal cognitive function assessment table (MoCA) score, serum prostate specific antigen (PSA) levels and 12-month survival rate were observed in 2 groups. RESULTS: The total response rate of observation group was significantly higher than that of control group, with statistical significance (P<0. 05). Before treatment, there was no statistical significance in MoCA score and serum PSA levels between 2 groups (P>0. 05). After treatment, MoCA scores of 2 groups were decreased significantly, and the observation group was higher than the control group, with statistical significance (P<0. 05). 6 and 12 months after treatment, serum PSA levels of 2 groups were decreased significantly, and the observation group was significantly lower than the control group, with statistical significance (P<0. 05); 12-month survival rate of observation group (92. 86%) was significantly higher than that of control group (64. 29%), with statistical significance (P< 0. 05). CONCLUSIONS: Nonsteroidal anti-androgen drugs show significant therapeutic efficacy for elderly prostate cancer, reduce cognitive function damage, improve serum PSA levels, therapeutic efficacy and short-term survival rate.
10.Comparison of plasma microRNA-1 and cardiac troponin T in early diagnosis of patients with acute myocardial infarction
Li-Ming LI ; Wen-Bo CAI ; Qin YE ; Jian-Min LIU ; Xin LI ; Xiao-Xing LIAO
World Journal of Emergency Medicine 2014;5(3):182-186
BACKGROUND: Early reperfusion can effectively treat acute myocardial infarction (AMI) and reduce the mortality significantly. This study aimed to compare the role of plasma microRNA-1 (miR-1) and cardiac troponin T (cTnT) in early diagnosis of AMI patients. METHODS: From May 2011 to May 2012, plasma samples were collected from 56 AMI patients and 28 non-AMI controls. The expression of plasma miR-1 was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and the level of plasma cTnT was measured using electrochemiluminescence-based methods on an Elecsys 2010 Immunoassay Analyzer. SPSS 16.0 was used for the statistical analysis of the results. Data were expressed as mean±standard deviation unless otherwise described. The differences about clinical characteristics between the AMI patients and controls were tested using Student'st test or Fisher's exact test. The Mann-WhitneyU test was conducted to compare the expression of microRNAs between the AMI patients and controls. MicroRNAs expression between different intervals of the AMI patients was compared using Wilcoxon's signed-rank test. The receiver operating characteristic (ROC) curve was established to discriminate the AMI patients from the controls. RESULTS: In the present study, the expression of plasma miR-1 was significantly increased in the AMI patients compared with the healthy controls (P<0.01). The plasma miR-1 in the AMI patients decreased to the normal level at 14 days (P>0.05). The expression of plasma miR-1 was not related to the clinical characteristics of the study population (P>0.05). ROC curve analyses demonstrated that miR-1 was specific and sensitive for the early diagnosis of AMI, but not superior to cTnT. CONCLUSION: Plasma miR-1 could be used in the early diagnosis of AMI, but it is similar to cTnT.