1.Study on p38 mitogen activated protein kinase in vascular endothelial cells dysfunction in preeclampsia
Xin LUO ; Dandan LIU ; Hongbo QI ; Zhenwei YAO
Chinese Journal of Obstetrics and Gynecology 2011;46(1):36-40
Objective To study expression and activation of p38 mitogen activated protein kinase (MAPK) in vascular endothelial cells dysfunction in preeclampsia. Methods From Sept. 2009 to Mar.2010, 54 pregnant women underwent deliveries in the First Affiliated Hospital of Chongqing Medical University were enrolled in this study, including 20 patients in mild preeclampsia group, 16 patients in severe preeclampsia group and 18 women with term cesarean section without perinatal complications as control group. Placental endothelial cells were labeled by CD34 to assay microvessel density (MVD) of each group. Immunohistochemical SP and western blot were used to detect localization and expression of p-p38 MAPK protein, respectively. The levels of sera soluble fms-like tyrosine kinase-1 (sFlt-1)and soluble endoglin(sEng) were measured by ELISA. Results ①The MVD of placenta were 103 ± 3 in control group, 81 ±5 in mild preeclampsia group and 63±4 in severe group, respectively, which showed statistical difference among each group (P<0.05).②The expression of p38 MAPK protein were 0.84±0.05 in control group,0.90±0.14 in mild group and 0. 86 ±0.18 in severe group, which did not reach remarkable difference among each group (P>0.05). The expression of p-p38 MAPK protein were 0.13±0.05 in control group,0.59±0.12 in mild group and 1.16±0.18 in severe group, which show statistical difference among each group(P<0.05).(3) The localization of p-p38 was in trophoblast, endothelial cells and a few (5.2±0.3)and(10.9±0.4)μg/L in control group,(12.5±1.2) and (20.4±5.3)μg/L in mild group and (19.3±3.0) and (29. 5 ±3.7) μg/L in severe group. When drawing paired comparison in those p-p38 MAPK protein levels and the concentrations of serum sFlt-1, sEng in preeclampsia groups (r=0.68,P<0.05;r=0.87,P<0.05). Conclusions The remarkable activation of the p38 MAPK in the placenta of patients with preeclampsia induced the increased levels of sFlt-1 and sEng in maternal serum, which confer the injury of vascular endothelial cells that caused the significant decline of MVD in placentas. p38 MAPK signaling might be one of the key pathways in vascular endothelial cell dysfunction in preeclampsia.
2.Content Determination of Hordenine in Zang Medicine Herba Aconiti by HPLC
Dashuang QING ; Xin WANG ; Yao QIN ; Lincai YAO ; Siping WANG ; Yong YANG ; Weizao LUO
China Pharmacy 2016;27(3):367-369
OBJECTIVE:To establish a method for the content determination of hordenine in Zang medicine Herba Aconiti. METHODS:HPLC was performed on the column of Gemini-NX C18 with mobile phase of acetonitrile-0.25 mol/l Ammonium ace-tate solution(ammonia water to adjust the pH to 9.5,gradient elution,at flow rate of 1 ml/min,the detection wavelength was 230 nm,the column temperature was 30 ℃,and the injection volume was 20 μl.RESULTS:The linear range of hordenine was 3.276-819μg/ml(r=0.999 5);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recovery was 96.21%-104.04%(RSD=1.23%,n=9). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the content determi-nation of hordenine in Zang medicine Herba Aconiti.
3.Isolation of human umbilical cord mesenchymal stem cells and differentiation into adipocytes and osteblasts
Shaoqing HE ; Zhenyu LUO ; Qiuying LIU ; Xiangrong ZHOU ; Mingquan DENG ; Xin LUO ; Runsi YAO ; Zhi GAO ; Yifei WANG
Chinese Journal of Tissue Engineering Research 2010;14(14):2492-2496
BACKGROUND:Culture condition,isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells,which lack of unified identification standards.Therefore,it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem calls(hUCMSCs).OBJECTIVE:To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts.METHODS:The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase.The morphology,proliferation and immunophenotype of the 3rd passage cells were analyzed,and then cells were induced to osteogenic and adipogenic differentiation in vitro.RESULTS AND CONCLUSION:The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro.Flow cytometry analysis revealed that the hUCMSCs were positive for CD29 CD44,CD59,CD105,but were negative for CD40,CD86 and HLA-DR.These calls could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions.The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro.This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.
5.The comparison of efficacy of cisplatin and nedaplatin for concurrent chemoradiotherapy for patients with locallyadvanced cervical cancer
Juan FU ; Aiyun ZHANG ; Xin YANG ; Aihua LUO ; Xia YAO ; Yifa YIN
Journal of Chinese Physician 2015;17(6):807-810
Objective To investigate the clinical effect of cisplatin and nedaplatin through comparing the effects of synchronous radiotherapy and chemotherapy of cisplatin and nedaplatin on patients with locally advanced cervical cancers.Methods Clinical data of 54 cases of patients with locally advanced cervical cancers received treatment at our hospital from 2009 to 2011 were analyzed retrospectively.Patients were divided into two groups according to the treatment:group DDP (cisplatin) and group NDP (nedaplatin).The general information,short-term effect,long-term curative effect,and adverse reactions were compared between two groups.Results The general information including gender,age,body mass index (BMI),stages,and pathology had no statistically significant difference between two groups.The response rate (RR)of patients in each group was 100%.The complete remission (CR) rate between groups NDP and DDP had no statistically significant difference.Among the patients ≤60 years old,the CR rate,the survival rate of 1,2,and 3 years,and the median progression-free survival (PFS) had no statistically significant difference.Among the patients > 60 years old,the survival rate of 1,2,and 3 years,and the median PFS had no statistically significant difference,but the CR rate had statistically significant difference.In the non-hematological toxicity,the incidence rate of nausea and vomiting,the effect on renal function in group NDP were obviously lower than those in group DDP.There was no significant difference in liver function damage.In the hematological toxicity,there was no statistically significant difference in the incidence rate of anemia and leukopenia.The incidence rate of decrease of platelet in group NDP was higher than that in group DDP,but it was mainly reflected in the Ⅰ and the Ⅱ level.Conclusions Cisplatin had the same efficacy of concurrent chemoradiotherapy with nedaplatin for the treatment of locally advanced cervical carcinoma with a worse tolerance.
6.Determination of Six Alkaloids in Crude and ProcessedRhizomaCoptidisby HPLC
Yong YANG ; Weizao LUO ; Jianbin SUN ; Xin WANG ; Lei HUA ; Longyun LI ; Yao QIN
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):596-602
An HPLC method to determine six alkaloids of the Coptidis for Chinese Pharmacopoeia of 2015 Edition was established through C18 column. The mobile phase was CH3CN-0.25 mol·L-1 NH4Ac (36:64) (containing 8 mmol·L-1 SDS and adjusting pH 9.3 with ammonia) at the flow rate of 1 mL·min-1, the detective wavelength was 270 nm and the column temperature was 35 oC. The linear ranges of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride and berberine hydrochloride were 0.006 96-0.233, 0.004 75-0.152, 0.003 30-0.528, 0.006 31-1.010, 0.004 71-0.753, 0.017 8-2.884 μg·mL-1, respectively. The average recoveries were 99.65%, 98.59%, 98.49%, 98.66%, 98.64%, 98.63% and RSD were 0.03%、0.15%、0.21%、0.12%、0.28%、0.23%, respectively. The method is simple and accurate, and can be used to determine the contents of jatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in the Coptidis.
7.Screening on Active Sites and the Best Compatibility of NewYouguiyin Prescription
Yao QIN ; Ziwei LI ; Dejiang WANG ; Xin WANG ; Qiang FU ; Weizao LUO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(5):985-989
This study was aimed to clarify the main active site and the best compatibility of the newYouguiyin prescription. The rat osteoblast-like cells was used as model cells. The investigation was made on the impact of different extraction parts of each herb and their compatibility on osteoblast proliferation. The results showed that the compatibility of butyl alcohol part fromDipsacus,Epimedium andCnidium had the most significant proliferation of osteoblast activity. It was concluded that the screening of active sites and the best compatibility of newYouguiyin prescription had laid the foundation for the development of subsequent innovation for traditional Chinese medicine (TCM).
8.Study on disulfide bond formation protein A in Escherichia coli.
Man LUO ; Yi-Xin GUAN ; Shan-Jing YAO
Chinese Journal of Biotechnology 2007;23(1):7-15
Disulfide bond formation protein A, DsbA, is one of the important proteins located in E. coli periplasm, which is a foldase facilitating the folding of nascent secreted proteins, especially for those with many pairs of disulfide bonds. The crystal structure and phylogenetic analysis of DsbA and DsbA-mediated protein folding, alternatively in vivo and in vitro, are summarized. Both the extremely low pKa of Cys30 , about 3.5, and the destabilizing effect of the active site disulfide contribute to its strong oxidizing power. The Cys30 is also considered as the most important residue closely related to its activity using site-directed mutagenesis methodology. DsbA could effectively assist proteins folding, both in vivo coexpressed with the target protein, and in vitro replenished as foldases. Moreover, DsbA also has the chaperone-like activity in the assistant refolding of genetically engineered inclusion bodies.
Amino Acid Sequence
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Disulfides
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chemistry
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metabolism
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Escherichia coli
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enzymology
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genetics
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Escherichia coli Proteins
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chemistry
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classification
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metabolism
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Models, Molecular
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Molecular Sequence Data
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Phylogeny
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Protein Disulfide-Isomerases
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chemistry
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classification
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metabolism
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Protein Folding
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Protein Structure, Tertiary
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Sequence Homology, Amino Acid
9.Organization and practice of hospital accreditation
Yutong WU ; Jian ZHANG ; Xiangping WANG ; Lihong WANG ; Xiaoying LI ; Zheng YAO ; Xin YANG ; Xiaoan WANG ; Tao LUO
Chinese Journal of Hospital Administration 2012;(11):804-807
The authors introduced,against the backdrop of the new round of accreditation,organization and practice of the hospital.In accordance with the five management elements of planning,organization,leadership,coordination and control,and the level management theory,the hospital divided,based on a top-down design and step-by-step implementation,the process into four stages of mobilization and deployment,study and training,self-assessment and rectification,supervised self-assessment and constant improvement.These efforts aim at exploring the key points and methodology of hospital accreditation,proposing such key points as the combination of the accreditation with building a long-term mechanism,that of theory with practice,that leadership with full staff involvement,that of top-down design with step-by-step implementation,that of training and rectification,that of self-assessment and supervision,and that of system management with implementation of provisions.This way the hospital accreditation may upgrade the hospital as a whole.
10.Identification of gentianae macrophyllae radix using the ITS2 barcodes.
Kun LUO ; Pei MA ; Hui YAO ; Tianyi XIN ; Yan HU ; Sihao ZHENG ; Linfang HUANG ; Jun LIU ; Jingyuan SONG
Acta Pharmaceutica Sinica 2012;47(12):1710-7
DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.