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1.
Chinese Journal of School Health ; (12): 1215-1219, 2022.
Article in Chinese | WPRIM | ID: wpr-940258

ABSTRACT

Objective@#To explore correlations between physical activity (PA), sedentary behavior (SB) and physical posture disorders in primary school students, and to provide reference for the prevention and correction of adverse body posture in this group.@*Methods@#From September to December 2020, 206 children from 4 primary schools in Chaoyang District of Beijing were selected by multistage stratified cluster random sampling method. Body posture and PA were assessed using a body posture tester and a triaxial accelerometer. Children s Leisure Activity Questionnaire was used to investigate SB.@*Results@#The detection rate of abnormal body posture in primary school students was 76.70%. PA showed positive assocation with high and low shoulder, X/O leg, pelvis forward, spine curvature and comprehensive evaluation ( β =0.17, 0.21, 0.19, 0.24, 0.19, P <0.05). SB had significant negative linear correlation with high and low shoulder, X/O leg,pelvis forward, spine curvature and comprehensive evaluation ( β = -0.24 , -0.22, -0.36, -0.24, -0.27, P <0.05). In the combination analysis, children with high PA plus low SB ( OR=2.12, 95%CI= 1.46-3.37, P <0.05) and children with low PA plus high SB had the highest and lowest advantages of qualified rate of body posture assessment,respectively. Compare with low SB and low PA( OR=1.71, 95%CI=1.41-2.74, P <0.05), high PA and high SB ( OR=1.85, 95%CI=1.38-2.86, P <0.05) had higher qualified rate of body posture assessment.@*Conclusion@#Physical activity shows positive effects on physical posture while sedentary behavior shows negative effects on physical posture. The combination of PA and SB has a counteracting or synergistic interaction effect.

2.
J Genet ; 2020 Jun; 99: 1-6
Article | IMSEAR | ID: sea-215507

ABSTRACT

Droughtmaster is a tropical breed of beef cattle that can survive in hot climates and easily adapt to torrid environments. These traits are important in livestock breeding. In this study, we genotyped five single-nucleotide polymorphisms (SNPs) of the AHSA2 gene from 190 cattle belonging to three different breeds (Droughtmaster, Angus and Simmental) by using snapshot technology. This work aimed to identify the valuable molecular marker of heat resistance in cattle. Results showed that Droughtmaster exhibited higher expected heterozygosity and polymorphic information content compared with the two other breeds. The AHSA2-1 locus deviated from the Hardy– Weinberg equilibrium in the Droughtmaster breed (P \ 0.05). Two SNPs in Droughtmaster diverged significantly from Angus and Simmental. The SNPs were identified as AHSA2-3 and AHSA2-4, which were closely linked to the three breeds based on pair-wise FST. AHSA2-4 involved a missense mutation. In summary, the GG genotypes in AHSA2-3 and AHSA2-4 may be candidate genotypes associated with heat resistance traits and may serve as valuable genetic markers for breeding of heat-tolerant beef cattle in the future.

3.
Chinese Journal of School Health ; (12): 1151-1154, 2020.
Article in Chinese | WPRIM | ID: wpr-825071

ABSTRACT

Objective@#BMI-based Silhouette Matching Test was used to quantify body image perception and expectation perception of female college students, and to investigate the predictive effect of body image perception deviation on dieting tendency. so as to provide evidence for further intervention measures.@*Methods@#A total of 579 female college students from 5 classes in two normal universities in Chongqing were conveniently sampled, and a questionnaire survey was conducted among the female college students. Multivariate stepwise regression was used to analyze the predictors of dieting tendency.@*Results@#About 41.62% of female college students had deviation between subjective somatotype perception and actual somatotype, and 81.69%(473) of female college students had deviation in their somatotype expectation after quantification. The degree of cognitive deviation of somatotype expectation was negatively correlated with eating behavior and positively correlated with dieting tendency(r=-0.10, 0.36, P<0.05). Dieting tendency was positively correlated with BMI and negatively correlated with eating behavior(r=0.14, -0.18, P<0.05). The degree of cognitive deviation of somatotype expectation, eating behavior, BMI, eating health awareness, and grade all had predictive effects on dieting tendency. The multivariate correlation coefficient between the five predictive variables and dieting tendency was 0.43, and the determination coefficient was 0.18. The overall test value of the regression model was 4.24(P=0.04). The five predictive variables accounted for 18.1% of the dieting tendency, among which the most powerful was the degree of cognitive deviation in somatotype expectation, which accounted for 12.9% of the variance.@*Conclusion@#There is a general deviation in female college students’ cognition of somatotype, mainly because they expect to be thinner. Irrational expectations of somatotype will lead to poor diet and even dieting behaviors, which will seriously affect their physical and mental health. Targeted intervention measures should be taken.

4.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 510-517, 2019.
Article in Chinese | WPRIM | ID: wpr-817668

ABSTRACT

@#【Objective】 A human embryonic stem cell line derived from Preimplantation genetic testing (PGT) embryos was established in a xeno- free stem cell culture system to provide disease models for medical research. 【Methods】The xeno-free culture system using xeno-free human foreskin fibroblast feeder layers(XF-HFF)mixed with commercially available chemically-defined medium(CDM)was assessed. In the culture system,a new hESC cell line was established using discarded embryos derived from PGT in patients with chromosomal balance translocation.【Results】The new availabled stem cell line was successfully cultured in the xeno-free culture system for a long time(> 45 passages). The karyotype analysis revealed that the new line kept the same karyotype over 45 passages. Moreover,the expression of pluripotent markers was detected by fluorescent immunostaining including SSEA- 3,SSEA- 4,SSEA- 1,TRA- 1- 60, and TRA-1-81. RT-PCR analysis showed that the stem cell markers were present in hESC grown on XF-HFF-CDM. In addition,the teratoma formation analysis demonstrated that the cells cultured in XF-HFF/CDM maintained their pluripotency in vivo.【Conclusions】Our study may provide the possibility to establish embryonic stem cells with certain pathogenic genes,which could be applied for clinical research and treatment.

5.
Chinese Journal of Pathophysiology ; (12): 403-409, 2018.
Article in Chinese | WPRIM | ID: wpr-701135

ABSTRACT

AIM:To observe the effects of transection of right cervical sympathetic trunk(TCST)on inflam-matory response and expression of high mobility group box 1(HMGB1)and TLR4/NF-κB signaling pathway in the rats af-ter acute myocardial infarction(AMI).METHODS: AMI model was established by ligation of left anterior descending coronary artery in SD rats,then the model rats were randomly divided into MI group and MI +TCST group.MI+TCST model was performed by transection of right cervical sympathetic trunk after left anterior descending coronary artery ligation. The rats in MI group and MI+TCST group were divided into 1,3,7,14 and 28 d subgroups,and another sham operation group threading without ligation, with 8 rats in above each group.After modeling for 4 weeks, the cardiac function was measured by echocardiography.All rats were killed to harvest the hearts for mesuring cardiac hypertrophy index.The myo-cardial tissue close to infarction was observed with HE staining.The relative mRNA expression levels of HMGB1, tumor necrosis factor α(TNF-α)and interleukin(IL)-6 at different time points were detected by real-time PCR.The protein ex-pression of HMGB1 and TLR4 at different time points after AMI was determined by Western blot.The effect of transection of right cervical sympathetic trunk on the expressions of HMGB 1 and TLR4/NF-κB signaling pathway was also analyzed. RESULTS:Compared with the MI group,left ventricular ejection fraction(LVEF)and left ventricular shorterning fraction (LVFS)were significantly higher(P<0.05),left ventricular end-diastole dimension(LVEDd),left ventricular end-sys-tole dimension(LVESd)and cardiac hypertrophy index were significantly lower(P<0.05), and the mRNA levels of HMGB1,TNF-αand IL-6 decreased significantly in MI +TCST group(P<0.05).Western blot results revealed that the protein expression level of HMGB1 increased in the infarct border zone at 3 d,and reached its peak at 7 d,then gradually decreased,and at 28 d after MI in MI group was still significantly higher than that in sham group(P<0.05).The protein expression of TLR4 was consistent with that of HMGB1.Transection of right cervical sympathetic trunk reduced protein ex-pression of HMGB1 and TLR4/NF-κB signaling pathway-related proteins(P<0.05).CONCLUSION: Transection of right cervical sympathetic trunk improves ventricular remodeling and maintaining cardiac function.The mechanism may be related to inhibiting HMGB1/TLR4/NF-κB signaling pathway to reduce inflammatory response.

6.
Chinese Journal of Anesthesiology ; (12): 863-865, 2018.
Article in Chinese | WPRIM | ID: wpr-709889

ABSTRACT

Objective To evaluate the efficacy of bronchial blocker outside the endotracheal tube for one-lung ventilation (OLV) in pediatric patients. Methods Thirty-eight American Society of Anesthe-siologists physical status ⅠorⅡ patients of both sexes, aged 4-8 yr, weighing 14. 0-28. 6 kg, scheduled for elective thoracic surgery under general anesthesia, were enrolled in this study. The size of tracheal tube was selected according to the endotracheal diameter at the level of clavicle and sternum on chest X-ray. Af-ter anesthesia induction, the distal end of the bronchial blocker was bent toward the target bronchus with a videolaryngoscope and then inserted until resistance was felt, and the preselected tracheal tube was inserted with a videolaryngoscope at a depth of (age∕2+12) cm. The position of the bronchial blocker was adjusted using a bronchofiberscope so that the upper edge of the blue cuff was 0. 5-1. 0 cm below the carina. The heart rate, mean arterial pressure, SpO2and airway pressure were recorded, arterial blood gas analysis was performed, and PaCO2and PaO2were recorded immediately before OLV (T1), at 30 min after OLV (T2) and at the end of OLV (T3). The intubation time, success rate of intubation at first attempt, displacement of bronchial blocker and occurrence of postoperative laryngeal pain and hoarseness were recorded. The pul-monary collapse was evaluated according to the standard described by Campos.Results Compared with the baseline at T1, PaO2was significantly decreased and airway pressure was increased at T2,3(P<0. 05), and no significant change was found in heart rate, mean arterial pressure, PaCO2or SpO2at T2,3( P>0. 05). The average intubation time was (185±38) s. The success rate of intubation at first attempt, rate of bronchial blocker displacement, excellent and good rates of pulmonary collapse and incidence of laryngeal pain and hoarseness were 95%, 16%, 100% and 11%, respectively. Conclusion Bronchial blocker outside the endotracheal tube can be effectively used for OLV in pediatric patients.

7.
China Journal of Chinese Materia Medica ; (24): 4582-4587, 2017.
Article in Chinese | WPRIM | ID: wpr-338234

ABSTRACT

To explore the resource of endophytic actinomycete in Fritillaria unibracteata, and alleviate the shortage of F. unibracteata resource, using F. unibracteata as experimental materials which growth in the western Sichuan plateau and cut its healthy bulb. Pure culture, insert, TLC and Oxford cup were applied to observe the mycelial morphology, research the ability of producing alkaloid and its antibacterial activity. Totally, 14 endophytic actinomycete strains were isolated by using Gao culture media. Based on the color reaction, 5 typical strains were selected for producing alkaloid. Through the TLC technique, all strains produced 2 obvious alkaloids spots. Antibacterial activity determination showed that the antimicrobial effects of 2 strains is prominent, the diameter up to 11 mm.16S rRNA gene sequence comparison analysis showed that 5 strains belonging to the Streptomyces. The alkaloids produced by endophytic actinomycetes are not related to F. unibracteata, but its fermentation liquid has antibacterial effect, it is worthy of further study.

8.
The Journal of Clinical Anesthesiology ; (12): 775-777, 2016.
Article in Chinese | WPRIM | ID: wpr-498186

ABSTRACT

Objective To compare the effect of different puncture positions in the combined spinal epidural anesthesia (CSEA)in cesarean section.Methods Ninety uterogestation women under-going elective Caesarean section were randomly divided into two groups,45 in each group.L3-4 inter-vertebral space was selected as puncture point.Group R kept right lateral position for two minutes af-ter puncture and turned 30°left tilt supine position until ready for surgery.Group L was turned to su-pine position after left lateral punctured and then turned to 30 left tilt supine position until ready for surgery.1% ropivacaine 1.5 ml+ 10% glucose 0.5 ml was used for spinal anesthesia.The sensory block level and adverse reaction within 1 5 min after the spinal anesthesia were observed,and the neo-natal Apgar score and the pH value of umbilical artery blood were recorded.Results The final block level of the group R was significantly lower than group L,and the time achieving final block height was significantly shorter than that in group L(P <0.05 ).The ephedrine dosage of group R 0 (0-6) mg was significantly lower than that of group L6 (0-12)mg (P <0.05).The PH value of umbilical artery blood in group R was significantly higher than that in group L (P <0.05).There was no sig-nificant difference in neonatal Apgar score.Conclusion In Cesarean section,keeping right lateral puncture position for two minutes after infusion and then turning 30° left tilt supine position can a-chieve better effects than 30°left tilt supine position immediately after puncturing.

9.
Journal of Medical Biomechanics ; (6): E181-E187, 2016.
Article in Chinese | WPRIM | ID: wpr-804026

ABSTRACT

The biomechanical studies on the temporomandibular Joint(TMJ)are of great importance to understand the origin and progression of TMD disorders. The finite element method can be used to analyze stress/strain distribution patterns in the TMJ tissues after application of force or deformation. However, due to the delicate and complicated anatomical structures of TMJ, modeling research were always time-consuming, expensive and non-repeatable. To explore a more rapid and accurate method for establishment of 3D finite element model of TMJ, the relevant factors such as geometric similarity, biomechanical environment simulation for improving the accuracy of TMJ finite element modeling were summarized.

10.
Chinese Journal of General Practitioners ; (6): 787-789, 2015.
Article in Chinese | WPRIM | ID: wpr-483094

ABSTRACT

The clinical data of 96 intubated inpatients with restricted cervical movement from June 2012 to June 2014 were retrospectively reviewed.They were divided into fiberoptic bronchoscope, Shikani optical stylet and McCoy laryngoscope groups (n =32 each).Identical anesthesia induction was applied.Compared with McCoy laryngoscope group, the total success rate and one-time success rate of tracheal intubation was higher in Shikani optical stylet group, tracheal intubation time was shorter and postoperative complications were fewer (P < 0.05).Compared with group F, endotracheal intubation time was shorter (P <0.05).However, heart rate, mean arterial pressure (MAP), oxygen saturation (SpO2), total success rate of tracheal intubation, one-time success rate of tracheal intubation and complication rate showed no significant inter-group differences at all timepoints (P > 0.05).Shikani optical stylet intubation has a less impact on hemodynamics and intubation time is short so that it is ideal for surgical patients with restricted cervical spine.

11.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 291-4, 2015.
Article in English | WPRIM | ID: wpr-637105

ABSTRACT

The reliability and validity of risk assessment scale (RAS) of pressure sore during 3S surgery were investigated. RAS of pressure sore was designed independently during 3S surgery. Five operating room nursing experts were selected to consult and detect face validity. Convenient and purposive sampling of 707 samples was conducted. Cronbach's alpha was used to measure content reliability and evaluate the internal consistence of RAS. The structural reliability was investigated by exploratory factor analysis method. The results showed that the content validity index was 0.92, and Cronbach's alpha of content reliability was 0.71. Structural validity, detected by Bartlett sphericity test, was 135.3 for 707 samples with the difference being statistically significant (P<0.01). KMO value was 0.729. The accumulative variance contribution ratio of common factor was 64.63%. The exploratory factor analysis showed the factor load of every clause was larger than 0.596. It was concluded that RAS of pressure sore for 3S surgery has better validity and reliability, and it could be used for evaluating and screening the high risk patients with pressure sores during surgery in order to efficiently reduce the occurrence of pressure sore during surgery. RAS of pressure sore for 3S surgery is worth to be popularized.

12.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 291-294, 2015.
Article in English | WPRIM | ID: wpr-331071

ABSTRACT

The reliability and validity of risk assessment scale (RAS) of pressure sore during 3S surgery were investigated. RAS of pressure sore was designed independently during 3S surgery. Five operating room nursing experts were selected to consult and detect face validity. Convenient and purposive sampling of 707 samples was conducted. Cronbach's alpha was used to measure content reliability and evaluate the internal consistence of RAS. The structural reliability was investigated by exploratory factor analysis method. The results showed that the content validity index was 0.92, and Cronbach's alpha of content reliability was 0.71. Structural validity, detected by Bartlett sphericity test, was 135.3 for 707 samples with the difference being statistically significant (P<0.01). KMO value was 0.729. The accumulative variance contribution ratio of common factor was 64.63%. The exploratory factor analysis showed the factor load of every clause was larger than 0.596. It was concluded that RAS of pressure sore for 3S surgery has better validity and reliability, and it could be used for evaluating and screening the high risk patients with pressure sores during surgery in order to efficiently reduce the occurrence of pressure sore during surgery. RAS of pressure sore for 3S surgery is worth to be popularized.


Subject(s)
Humans , Intraoperative Period , Pressure Ulcer , Reproducibility of Results , Risk Assessment , Surveys and Questionnaires
13.
Journal of Experimental Hematology ; (6): 142-145, 2015.
Article in Chinese | WPRIM | ID: wpr-259625

ABSTRACT

<p><b>OBJECTIVE</b>This study was to detect the plasma thrombomodulin (TM), D-dimer and fibrinogen in patients with multiple myeloma (MM) and to analyze their relationship with morbid state, and also to investigate the relationship of the expression of coagulation factor with the ratio of myeloma cells.</p><p><b>METHODS</b>ELISA was used to detect the TM level in 45 cases of MM at different stages. The plasma level of D-dimer and fibrinogen was detected by STA automatic coagulation analyser.</p><p><b>RESULTS</b>The level of plasma TM in newly diagnosed patients was higer than that in normal control group and in platform stage group (P < 0.01; P < 0.05). There were significant differences between relapsed or refractory group and normal control group or those reached platform stage group (P < 0.05). Meanwhile, the level of plasma TM in the group of thalidomide combined with chemotherapy was higer than that in newly diagnosed patients (P < 0.05). The level of plasma D-dimer and fibrinogen of MM patients was higher than that in normal controls (P < 0.01;P < 0.05). The expression of D-Dimer in relapsed or refractory group reached the maximum. Also, the level of plasma D-Dimer in group of thalidomide combined chemotherapy was higer than in newly diagnosed patients (P < 0.05). The expression of coagulation factor did not correlate with the ratio of myeloma cells.</p><p><b>CONCLUSIONS</b>Level of plasma TM, D-Dimer and fibrinogen of MM patients is higher than that in control group. The level of plasma TM and D-Dimer can be elevated when thalidomide used, which indirectly suggested the tendency for thrombosis in MM patients.</p>


Subject(s)
Humans , Blood Coagulation Tests , Fibrin Fibrinogen Degradation Products , Fibrinogen , Multiple Myeloma , Thalidomide , Thrombomodulin , Thrombosis
14.
São Paulo med. j ; 131(6): 440-440, 2013.
Article in English | LILACS | ID: lil-697429

ABSTRACT

BACKGROUND: It had been assumed that suppressing the undamaged contralesional motor cortex by repetitive low-frequency transcranial magnetic stimulation (rTMS) or increasing the excitability of the damaged hemisphere cortex by high-frequency rTMS will promote function recovery after stroke. OBJECTIVE: To assess the efficacy and safety of rTMS for improving function in people with stroke. METHODS: AUTHORS' CONCLUSIONS: Current evidence does not support the routine use of rTMS for the treatment of stroke. Further trials with larger sample sizes are needed to determine a suitable rTMS protocol and the long-term functional outcome. .


Subject(s)
Humans , Recovery of Function , Stroke/rehabilitation , Transcranial Magnetic Stimulation/methods
15.
Chinese Journal of Stomatology ; (12): 144-146, 2009.
Article in Chinese | WPRIM | ID: wpr-346717

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the manifestation of oral squamous cell carcinoma with metastatic lymph nodes in whole-body diffusion weighted imaging (WB-DWI) and its clinical significance.</p><p><b>METHODS</b>Twenty-one cases of oral squamous cell carcinoma with metastatic lymph nodes were examined by WB-DWI, of which 19 were scanned with routine MRI, 9 cases examined by positron-emission tomography (PET). All cases were confirmed by pathology. MRI scan covered whole body and built whole body diffusion image after reconstruction with GE HDe 1.5T MRI scanner.</p><p><b>RESULTS</b>There were 139 metastatic lymph nodes, 11 normal lymph nodes, and 21 inflammatory lymph nodes exhibited by WB-DWI in 21 cases. Metastatic lymph nodes showed higher signal and lower value of apparent diffusion coefficient (ADC). The mean ADC value of metastatic lymph nodes (0.78 +/- 0.07) was significantly lower than that of inflammatory (1.18 +/- 0.15) and normal nodes (1.78 +/- 0.16), and normal nodes showed even higher levels of ADC value.</p><p><b>CONCLUSIONS</b>STIR-EPI-DWI is a new promising technique for differentiating normal, inflammatory, and metastatic lymph nodes and can provide more useful information on lymph node metastasis.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Pathology , Lymph Nodes , Pathology , Lymphatic Metastasis , Pathology , Mouth Neoplasms , Pathology , Whole Body Imaging , Methods
16.
Journal of Experimental Hematology ; (6): 382-384, 2009.
Article in Chinese | WPRIM | ID: wpr-302128

ABSTRACT

In order to investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in patients with multiple myeloma and its significance, the expression of VCAM-1 in 23 patients were detected by reverse transcription-PCR (RT-PCR) and then compared with that in normal control. The results showed that the expression of VCAM-1 in patients with newly diagnosed and relapsed/refractory multiple myeloma was much higher than that in normal control and in patients reached plateau stage (p < 0.05). There was no statistically significant difference between newly diagnosed and relapsed/refractory patients (p > 0.05). It is concluded that the expression of VCAM-1 in patients with multiple myeloma is abnormal, which may be an important factor for the pathogenesis of multiple myeloma.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Case-Control Studies , Multiple Myeloma , Metabolism , Pathology , Neoplasm Staging , Reverse Transcriptase Polymerase Chain Reaction , Vascular Cell Adhesion Molecule-1 , Metabolism
17.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 379-382, 2008.
Article in Chinese | WPRIM | ID: wpr-260152

ABSTRACT

To investigate whether glutamate and voltage-gated calcium channels-independent calcium influx exists during acute anoxic neuronal damage and its possible relationship to neuronal protective function of NGF. In in vitro model of acute anoxia, hippocampal cultures from newborn rats were exposed to 3 mmol/L KCN. Changes of intracellular Ca2+ concentration ([Ca2+]i) were monitored by con-focal imaging and cell viability was assayed by PI and cFDA staining. The results showed that after treatment with primary hippocampal cultures with 3 mmol/L KCN for 15 min,[Ca2+]i was significantly increased 6.27-fold compared to pre-anoxia level and 73.3% of the cells died.When combination of 20 μmol/L MK-801 (glutamate receptor antagonist), 40 μmol/L CNQX (AMPA receptor antagonist) and 5 μmol/L nimodipine (voltage-gated calcium channel antagonist) (hereafter denoted as MCN) were administrated to hippocampal cultures, levels of [Ca2+]i and cell death rate induced by KCN were partially reduced by 35.9% and 47.5% respectively. However, Gd3+ (10μmol/L) almost completely blocked KCN-mediated [Ca2+]i elevation by 81.9% and reduced neuronal death by 88.8% in the presence of MCN. It is noteworthy that NGF, used in combination with MCN,inhibited KCN-induced [Ca2+]i increase by 77.4% and reduced cell death by 87.1%. Only PLC inhibitor U73122 (10 μmol/L) abolished NGF effects. It is concluded that Gd3+-sensitive calcium influx,which is NMDA (glutamate receptor) and voltage-gated calcium channels-independent, is responsible for acute anoxic neuronal death. NGF can inhibit Gd3+-sensitive calcium influx and reduce anoxic neuronal death through activating PLC pathway.

18.
Acta Academiae Medicinae Sinicae ; (6): 42-47, 2005.
Article in Chinese | WPRIM | ID: wpr-343769

ABSTRACT

<p><b>OBJECTIVE</b>To study the expression of human myofibrillogenesis regulator 1 (MR-1) gene in E. coli and obtain the MR-1 protein and its antibody for further investigation of its biological function.</p><p><b>METHODS</b>Expression vectors pGEX-5X-1, pET30a (+), and pET24a (+), as well as host strain E. coli BL21 (DE3) and BL21-CodonPlus (DE3) -RIL were used for expression of MR-1. MR-1 N-terminal with GST or T7-tag or C-terminal with His-tag, separately, or N terminal with T7-tag and C terminal with His-tag, simultaneously, were fused in plasmids pGEX-5X-1, pET30a (+) , and pET24a (+). The expressed MR-1-T protein, separated and purified by preparative SDS-PAGE, was applied to immunize the rabbits. The titer of the antibody was assayed by ELISA and its immunogenicity was tested by Western blot with pcDNA3/MR-1 transfected human breast cancer cell MCF7.</p><p><b>RESULTS</b>The MR-1 protein was successfully expressed as inclusion body by fusing its N-terminal with T7-tag in E. coli BL21-CodonPlus (DE3) -RIL. MR-1 protein was purified by electro-elution from SDS-PAGE gel. Using this purified protein, polyclonal antibody in rabbit against MR-1 was essentially generated. ELISA and Western blot showed the titer of this antibody was about 1:10(5) with high immunogenicity.</p><p><b>CONCLUSIONS</b>The N-terminal fusion tag is the most important mechanism for MR-1 expression. The polyclonal antibody of the generated MR-1 protein in E. coli may be applied for its further biological function studies.</p>


Subject(s)
Animals , Female , Humans , Rabbits , Antibodies , Breast Neoplasms , Metabolism , Pathology , Cell Line, Tumor , Escherichia coli , Metabolism , Genetic Vectors , Immunization , Muscle Proteins , Genetics , Allergy and Immunology , Plasmids , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Transfection
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