1.Culture, identification of phenotype,and labeling of mesenchymal stem cells in vitro in SD rats
Yong ZHU ; Liangwan CHEN ; Ruobai LIN ; Ziyang HAN ; Mingqiang KANG
Journal of Jilin University(Medicine Edition) 2006;0(02):-
Objective To investigate the methods of isolation,culture,identification and labeling of mesenchymal stem cells(MSCs) in vitro and lay a foundation for further study on intervention of MSCs on immunologic rejection of organ transplantation. Methods MSCs were isolated and cultivated by adherent methods . The expressions of CD90 and CD45 of cells were analyzed by using flow cytometry in order to identify MSCs.The third generation of MSCs were labeled by DAPI,the labeling efficiency was detected.Results Primary cultured MSCs adhered to plastic surface within 48 h and reached 90% confluence within 7-10 d .Flow cytometry showed that the positive rates of CD90 and CD45 of MSCs at third generation were 99.8% and 6.8%. MSCs expressed CD90 but no CD45.All of the MSCs after labeling by DAPI showed blue fluorescence by immunofluoroscope. DAPI labeling was sensitive and highly efficient to MSCs.Conclusion Adherent method is simple and easy to isolate and cultivate MSCs and it can serve as a routine method.DAPI labeling can be used as a efficient method to label MSCs.
2.The influence of specimen storage time to platelet count in unpaid blood donors of platelets
Ziyang FENG ; Lishuang ZHU ; Heshan TANG ; Baohua QIAN
Chinese Journal of Blood Transfusion 2017;30(7):704-705
Objective To analyse the effect of specimen storage time on platelet count in apheresis Donors,in order to choose the appropriate testing time.Methods We choose fifty healthy Unpaid Blood Donors of Platelets to test platelet count in 0,0.5,1,3,6h respectively by the blood counting instrument.Results At room temperature,the count of platelets from blood samples is relatively lower at 0 hour and the difference is significant (P < 0.05).After 0.5-6 hours,the count of platelets become stabilized and not has significantly different(P>0.05).Conclusion The count of platelets of blood samples is lower at 0 hour than 0.5-6 hours,this work suggest that the count of platelets of blood samples should be done at 0.5-6 h in order to protect platelets quality.
3.Comparison of Clinical Efficacy of Lyophilized Recombinant Human Brain Natriuretic Peptide and Dobutamine in Treatment of Acute Heart Failure and Plasma Gal-3, CysC and ET-1 Levels
Zhen ZHU ; Jianrong ZHAO ; Ziyang LI ; Qiang LU ; Hong SHAO
Progress in Modern Biomedicine 2017;17(26):5145-5148,5156
Objective:To explore and compare the clinical efficacy of lyophilized recombinant human brain natriuretic peptide (Lrh-BNP) and dobutamine (Dob) in the treatment of patients with acute heart failure (AHF) and impacts on the plasma galectin (Gal)-3,Cystatin C (CysC) and endothelin (ET-)-1 levels.Methods:114 cases of patients with AHF in our hospital from February 2015 to February 2017 were selected as the research objectives and randomly divided into two groups.Dob group was treated by Dob,while Lrh-BNP group was treated by Lrh-BNP.The cardiac function parameters,plasma Gal-3,CysC,ET-1 levels before and after treatment,clinical comprehensive efficacy and incidence of adverse reactions were compared between two groups.Results:The FS,LVEF levels of both groups at 72 hours after treatment were significantly higher than those before treatment (P<0.01),but the LVEDD,plasma Gal-3,CysC,ET-1 levels were obviously decreased (P<0.01),the index mentioned above of Lrh-BNP group improved more significantly than those of the Dob group(P<0.01).The overall effective rate of Lrh-BNP group was 89.5 %,which was significantly higher than that of the Dob group (73.7%,P<0.05).No significant difference was found in the incidence of adverse reaction between two groups(P>0.05).Conclusion:Lyophilized recombinant human brain natriuretic peptide was more effective in the treatment of AHF than Dobutamine with equal safety,which might be related to the decrease of plasma Gal-3,CysC,ET-1 levels.
4.Clinical analysis of Prader-Willi syndrome in 10 children
Qiaoli ZHOU ; Bei HAN ; Ziyang ZHU ; Wei GU ; Qianqi LIU ; Xing SHI ; Shining NI
Chinese Journal of Applied Clinical Pediatrics 2016;31(20):1578-1579
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5.Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS
Keren WANG ; Hai ZHANG ; Zhenyu ZHU ; Ziyang LOU ; Liang ZHAO ; Yifeng CHAI
Journal of Pharmaceutical Analysis 2010;22(3):149-155
A global quality control method based on high performance liquid chromatography (HPLC) coupled with diode array detection (DAD), single quadrupole mass spectrometry (MS) and time-of-flight mass spectrometry (TOFMS) was developed for simultaneous determination of seven major components (mangiferin, neomangiferin, timosaponin E1, timosaponin E, timosaponin BⅡ, timosaponin BⅢ, and timosaponin AⅢ) and identification of most components in extracts of Rhizoma Anemarrhenae (RA). HPLC analysis was performed on an Agilent SB-C18 column (4.6 mm×150 mm, 5 μm) by gradient elution using acetonitrile and water-acetic acid(100∶0.05, v/v) as the mobile phase. Seven major components in RA were successfully separated. This quantitative method was fully validated in respect of the following performance criteria: linearity, precision, repeatability, stability, accuracy, limits of detection (LOD) and quantification (LOQ). A formula database of known compounds in RA was established, against which, most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS. This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China. This global quality control method, which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components, is suitable for routine quantification and comprehensive quality control of RA.
6.Comparative analysis of essential oils found in Rhizomes Curcumae and Radix Curcumae by gas chromatography-mass spectrometry
Diya LU ; Yan CAO ; Ling LI ; Zhenyu ZHU ; Xin DONG ; Hai ZHANG ; Yifeng CHAI ; Ziyang LOU
Journal of Pharmaceutical Analysis 2011;01(3):203-207
A comparison of the volatile compounds in Rhizomes Curcumae (Ezhu) and Radix Curcumae (Yujin) was undertaken using gas chromatography mass spectrometi-y (GC-MS). Ultrasonic extraction and GC-MS methods were developed for the simultaneous determination of five sesquiterpenes, namely, α-pinene, β-elemene, curcumol, germacrone and curdione, in Ezhu and Yunjin. Good linearity (r〉0.999) and high inter-day precision were observed over the investigated concentration ranges. The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin. The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.
7.Etiology and bone age of 2132 children with short stature
Su WU ; Sumei WANG ; Ziyang ZHU ; Wei GU ; Shining NI ; Xing SHI ; Qianqi LIU
Journal of Clinical Pediatrics 2015;(8):730-733
ObjectiveThe aim of this study is to analyze the etiology and status of bone age of children with short stat-ure.MethodsAnthropological and physical examination data were retrospectively collected and studied in 2132 children with short stature in the department of endocrinology between 2009 and 2014. Growth hormone (GH) levels were determined by ar-ginine-clonidine test. Bone age was determined by CHN scoring.ResultsAmong the 2132 patients, 1333 were males and 799 were females. Mean age is 9.03 ± 3.04 years old, mean bone age is 6.81 ± 3.05 years. Of them, 324 cases (15.2%) were diagnosed complete GH deifciency, 780 cases (36.59%) were partial GH deifciency, 27cases (1.27%) were multiple pituitary hormone de-ifciency, 13 cases (1.64%) were hypothyroidism, 893 cases (41.89%) were idiopathic short stature, 19 cases (0.89%) were small for gestational age (SGA), 40 cases (1.88%) were chromosomal disorders, etc. Signiifcant difference in age and bone age was found using t test (P<0.05). Signiifcant differences in Δage were found between etiological categories using ANOVA (P=0.000). Δage was signiifcantly and negatively associated with peak GH using Pearson's correlation.ConclusionsGH deifciency is the most common cause of short stature. Bone age of children with short stature is commonly delayed. Δage was signiifcantly and negatively associated with peak GH. Multiple pituitary hormone deifciency has a signiifcant effect on bone age. The etiology of patients with short stature cannot be determined just by bone age.
8.Comparative analysis of essential oils found in Rhizomes Curcumae and Radix Curcumae by gas chromatography-mass spectrometry
Diya Lü ; Yan CAO ; Ling LI ; Zhenyu ZHU ; Xin DONG ; Hai ZHANG ; Yifeng CHAI ; Ziyang LOU
Journal of Pharmaceutical Analysis 2011;01(3):203-207
A comparison of the volatile compounds in Rhizomes Curcumae (Ezhu) and Radix Curcumae (Yujin) was undertaken using gas chromatography-mass spectrometry (GC-MS).Ultrasonic extraction and GC-MS methods were developed for the simultaneous determination of five sesquiterpenes,namely,α-pinene,β-elemene,curcumol,germacrone and curdione,in Ezhu and Yunjin.Good linearity (r>0.999) and high inter-day precision were observed over the investigated concentration ranges.The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin.The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.
9.The expressions of notch genes in human keloid-derived mesenchymal-like stem cells.
Chengliang DENG ; Bo WANG ; Ziyang ZHANG ; Guangfeng SUN ; Jingjing ZHU ; Dali WANG ; Limei YU
Chinese Journal of Plastic Surgery 2014;30(3):197-202
OBJECTIVETo study the expressions of Notch1-4 gene in human keloid-derived mesenchymal-like stem cells, and to explore the Notch signaling pathway's role in the formation of keloid.
METHODSKeloid samples were collected to harvest human keloid-derived mesenchymal-like stem cells through two-step enzymatic dissociation method. By flow cytometry, cell phenotype of primary and P3 generation were analyzed. By immunocytochemistry, the expressions of Oct4, vimentin and CK19 were examined. Keloid-derived mesenchymal-like stem cells were induced into osteoblasts in vitro and calcium deposition was detected by Alizarin red S stain. Realtime polymerase chain reaction (RT-PCR) was used to detect the expressions of Notch1-4 mRNA in keloid-derived mesenchymal-like stem cells.
RESULTSFlow cytometry showed that keloid-derived mesenchymal-like stem cells of primary and P3 generation highly expressed CD29, CD44, CD90 from the typical MSC phenotype marker, but they failed to express HSC phenotype markers, such as CD34 and CD45. The results of immunocytochemistry showed that Oct4 from pluripotent stem cell markers and vimentin from mesenchymal cell markers was positive and CK19 from epithelial cell markers was negative. After induced differentiation into osteoblasts in vitro after 21 day, calcium nodules could be seen clearly; Notch1-4 gene were expressed in keloid-derived mesenchymal-like stem cells through RT-PCR. The relative quantitative of Notch2, Notch3 gene were higher than Notch1, Notch4 gene (P < 0.05).
CONCLUSIONSThe expression difference of different subtypes from Notch gene in human keloid-derived mesenchymal-like stem ceils may be related to self-renewal, proliferation, differentiation, and participate in the formation of keloid.
Adolescent ; Cells, Cultured ; Child ; Female ; Humans ; Keloid ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; Receptors, Notch ; metabolism
10.17 beta-hydroxysteroid dehydrogenase 3 deficiency due to novel compound heterozygous variants of HSD17B3 gene in a sib pair.
Su WU ; Bixia ZHENG ; Ting LIU ; Ziyang ZHU ; Wei GU ; Qianqi LIU
Chinese Journal of Medical Genetics 2021;38(8):787-790
OBJECTIVE:
To explore the genetic basis for a sib pair featuring 17beta-hydroxysteroid dehydrogenase type 3 deficiency.
METHODS:
Genomic DNA was extracted from the proband, her sister, and their parents, and was subjected to sequencing analysis with a gene panel for sexual development. Suspected variant was verified by Sanger sequencing and bioinformatic analysis.
RESULTS:
Both the proband and her sister were found to harbor novel compound heterozygous missense variants of the HSD17B3 gene, namely c.839T>C (p.Leu280Pro) and c.239G>T (p.Arg80Leu), which were derived respectively from their mother and father. The variants were unreported previously and predicted to be deleterious by PolyPhen2, MutationTaster and other online software. Based on the American College of Medical Genetics and Genomics standards and guidelines, both c.839T>C(p.Leu280Pro) and c.239G>T (p.Arg80Leu) were predicted to be likely pathogenic (PM2+PP1+PP2+PP3+PP4, PM2+PM5+PP1+PP2+PP3+PP4).
CONCLUSION
The compound heterogeneous variants of the HSD17B3 gene probably underlay the disease in this sib pair. 17beta-hydroxysteroid dehydrogenase type 3 deficiency may lack specific clinical features and laboratory index, genetic testing can facilitate a definitive diagnosis.
17-Hydroxysteroid Dehydrogenases/genetics*
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Female
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Genetic Testing
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Genomics
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Humans
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Mutation
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Mutation, Missense