1. Construction and characterization of hGM-CSF-expressing K-562 cell line
Academic Journal of Xi'an Jiaotong University 2007;19(2):200-203
Objective: The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods: The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results: We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion: The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.
2. Construction and characterization of hGM-CSF-expressing K-562 cell line
Academic Journal of Xi'an Jiaotong University 2007;19(2):200-203
Objective: The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods: The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3. 1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results: We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion: The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.
3.OBSERVATION OF CURATIVE EFFECT OF HA IN 23 CASES WITH CML
Yichao QIAO ; Yinxia CHEN ; Goudong TIAN
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(02):-
The 23 cases with chronic myelogenousleukemia (CML) were treated with program ofharringtonine and ara-c (HA) and the results werethat the 13 cases got complete remission and 10cases got partial remission. Comparing with thecontrol group the patients of which were treatedwith busulfan, the authors found that the spendedtime of decreased WBC in blood (WBC decreasedto 10. 0 ? 10~9/L) and reduced spleen (reduced tosmallest status of spleen) and improvement of self-sensation symptoms in the group treated with HAwere better than those in control group. The HAprogram for cases who recured CML again and re-ceived maintenance cure after remission of CMLwas still effective. The time of maintenance cureand interval should be individualized.
4.The enantioselective pharmacokinetic study of desvenlafaxine sustained release tablet in Chinese healthy male volunteers after oral administration.
Yinxia CHEN ; Jiangbo DU ; Yifan ZHANG ; Xiaoyan CHEN ; Dafang ZHONG
Acta Pharmaceutica Sinica 2015;50(4):486-91
A chiral LC-MS/MS method for the simultaneous analysis of desvenlafaxine (DVS) enantiomers in human plasma was developed and applied to a pharmacokinetic study on 12 Chinese healthy volunteers. d6-Desvenlafaxine was used as internal standard (IS). Chromatographic separation was performed on the Astec Chirobiotic V chiral column (150 mm x 4.6 mm, 5 μm). The assay was linear over the concentration range of 0.500-150 ng x mL(-1) for both enantiomers (r2 > 0.99). The method was successfully applied to a stereoselective pharmacokinetic study of 100 mg desvenlafaxine sustained release tablets on 12 Chinese healthy volunteers under fasting conditions. The results showed that the pharmacokinetic parameters were similar to both enantiomers in Chinese healthy volunteers. The AUC(0-t), and C(max) of the two enantiomers were about 1.5 times higher than those of blacks and whites reported in the literature.
5.Impact of electroacupuncture on the life quality in patients of female mixed urinary incontinence.
Yinxia CHEN ; Enhui HE ; Hongfang TIAN ; Jiping ZHAO
Chinese Acupuncture & Moxibustion 2016;36(3):256-258
OBJECTIVETo observe the impact of electroacupuncture (EA) on the quality of life in the patients of female mixed urinary incontinence (MUI) and evaluate its clinical efficacy.
METHODSEA was adopted in 25 cases of MUI at Zhongliao (BL 33) and Huiyang (BL 35), needle retaining for 30 min each time. The treatment was given once every two days, three treatments a week, totally for 12 weeks of treatment. The incontinence quality of life questionnaire (I-QOL), the incontinence impact questionnaire-7 (IIQ-7), the urogenital distress inventory short form (UDI-SF) and the visual analogue scale (VAS) were adopted in the comparison before and after treatment. The efficacy was evaluated.
RESULTSThe curative rate was 52.0% (13/25) and the total effective rate was 96.0% (24/25) in 25 cases. In comparison before and after treatment, in I-QOL, the score of limiting behaviors was increased up to (30.32 ± 3.39) from (11.52 ± 2.45), the score of psychosocial impacts was up to (36.48 ± 2.90) from (13.52 ± 2.25) and the score of social isolation was up to (21.52 ± 1.61) from (9.84 ± 2.23). The score of IIQ-7 was decreased, down to (2.48 ± 1.36) from (11.96 ± 3.45), the score of UDI-SF was down to (2.92 ± 0.95) from (8.84 ± 2.58) and VAS score was down to (1.40 ± 0.91) from (6.96 ± 1.65). The differences were all significant statistically in comparison (all P < 0.05).
CONCLUSIONEA improves effectively the quality of life in MUI patients and the clinical efficacy is apparent.
Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Middle Aged ; Quality of Life ; Treatment Outcome ; Urinary Incontinence ; physiopathology ; therapy ; Urination
6.Evaluation for clinical application effect of gene chip for detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis
Rongqing XU ; Dan LI ; Yinxia LIN ; Mingxiang HUANG ; Xinchao CHEN
Chinese Journal of Zoonoses 2017;33(1):43-48
We evaluated clinical application effect of gene chip for detection of rifampin (RFP) and isoniazid (INH) resistance in Mycobacterium tuberculosis (MTB).Rifampin and isoniazid drug-resistance gene loci were detected by gene chip with sputum specimens from smear-positive tuberculosis patients and clinical strains,comparing the results of detection.BACTEC MGIT 960 drug susceptibility test results were used as control to evaluate the detection performance of gene chip.The sequences of the polymerase chain reaction products of the rpoB,katG and inhA genes from 999 strains identified as Mycobacterium tuberculosis were determined to confirm the mutations by DNA sequencing.Results showed that 100 cases were identified as nontuberculous mycobacteria by gene chip in the 1 108 cases of smear-positive samples.Among the rest 1 008 samples,there were only 9 cases of microarray results different from BACTEC MGIT960 culture-positive strains,achieving the coincidences of 99.1%.Compared with BACTEC MGIT 960 drug susceptibility test results,the gene chip method displayed a concordance of 98.1 % and 94.5 % for RFP and INH respectively in the 999 strains.Compared with the DNA sequencing method,the accuracy of gene chip method was 99.6% for rifampin resistance and 99.8% for isoniazid resistance.It's concluded that the gene chip technology can quickly and accurately detect rifampin and isoniazid resistance in MTB and can be used directly for the detection of sputum samples.
7.Effect of the shenxiong glucose injection on the recovery of neural function and the expression of AQP-4 in rats after spinal cord injury
Yinxia WU ; Xiaoyan YUAN ; Xinsheng CHEN ; Shuyuan WANG ; Chengfang HUANG
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(12):907-912
Objective To explore the action mechanism of shenxiong glucose injection in treatment of acute spinal cord injury(SCI) through observing its effects on the recovery of motor function and the expression of aquaporin-4(AQP-4) in SCI rats.Methods Totally 90 healthy,aduh,Sprague-Dawley rats were randomly divided into a sham operation group (n=30),aSCI group (n=30) and a drug group (n=30).The SCI rat models in both the SCI group and the drup group were established aecording to the modified Allen's method,while the sham operation group was only given laminectomy.After the operation,the drug group was given intraperitoneal shenxiong glucose injection of 30 rnl/kg a day,while the other two groups were injected in the same way with normal saline.The neural function recovery,the pathological changes after SCI and the expressions of AQP-4 were observed 1,3,7,14 and 21 d after the operation using the Tarlov score,the hematoxylin and eosin staining,as well as immunofluorescence techniques and Western blotting.And the correlation of Tarlov scores with AQP-4 expressions was analyzed.Results No significant changes in Tarlov scores were observed in the sham operation group (P > 0.05),while in the SCI group and the drug group,postoperative Tarlov scores decreased significantly.The hindlimb nerve function recovered to some degree with time in the SCI group and drug group.At 3,7,14 and 21 days after the operation,the Tarlov scores in the drug group were significantly higher than the SCI group (P < 0.05).The drug group showed less severe pathological changes,with more residual neurons still visible of nucleoli than the SCI group 21 days after the operation.Compared with the sham operation group,the expression levels of AQP-4 were significantly higher in the SCI group and drug group at all the time points (P < 0.05).However,the expression levels of AQP-4 in the drug group were significantly lower than the SCI group accordingly (P < 0.05).The Tarlov scores were found to be significantly and negatively related to the AQP-4 protein expression levels 3 days(r =-0.523,P =0.003),7 days(r =-0.437,P=0.016),14 days(r=-0.417,P=0.022) and 21 days(r=-0.377,P=0.040)after the operation.Conclusion Injecting shenxiong with glucose can effectively promote the recovery of motor function after SCI,at least in rats.And its mechanism may be that the development of spinal cord edema is prevented and the secondary spinal cord injury alleviated by restraining the expressions of AQ P-4 in the injured areas.
8.Anticonvulsive action of Pinellia Pedatisecta Schott extract prepared by ethanol-modified supercritical CO2 extraction
Jingjing CHEN ; Rong YANG ; Mingzheng WANG ; Yinxia CHENG ; Xingu HE ; Yonggang MA ; Lihua YANG ; Qiaoyan HE
Chinese Journal of Pharmacology and Toxicology 2007;21(6):449-454
AIM To investigate the anticonvulsive action of supercritical CO2 ethanol extract from Pinellia Pedatisecta Schott(SEE-CO2PP). METHODS The rat convulsive model was induced by penicillin localized injected in rat cortex. The effects of SEE-CO2PP on the latency of seizure and changes of convulsive behaviors were investigated. The latency of epileptiform discharge, and frequency and amplitude of highest wave in cortex and hippocampus were recorded by using RM6240C multichannel physiological signal collection and analysis recorder. At the same time, the contents of glutamic acid (Glu), aspartic acid (Asp), glycine (Gly) and γ-aminobutyric acid (GABA) in hippocampus were determined with high performance liquid chromatography. RESULTS SEE-CO2PP 15 and 30 g·kg-1, ig, prolonged the latent period of seizure and weakened the extent. SEE-CO2PP also prolonged the latent period of epileptiform discharge, reduced the frequency and decreased amplitude of the highest wave in both cortex and hippocampus. Moreover, SEE-CO2PP increased the content of GABA in hippocampus, but the levels of Gly,Asp and Glu had no obvious changes. CONCLUSION SEE-CO2PP inhibits the epileptiform discharge and convulsive behaviors of convulsive model rats, which suggests SEE-CO2PP has anticonvulsive action.
10.Construction and characterization of hGM-CSF-expressing K-562 cell line
Yinxia CHEN ; Suhu LIU ; Yun YANG ; Jie LIU ; Xiaorong MA ; Wanggang ZHANG
Journal of Pharmaceutical Analysis 2007;19(2):200-203
Objective The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3.1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.