1.Expression and clinical significance of HA117 in adults with acute leukemia
Journal of International Oncology 2011;38(8):636-639
Objective To investigate the expression of HA117 in bone marrow mononuclear cells (BMMNCs) of acute leukemia patients, and its correlation with multidrug resistance( MDR), as well as its correlation with antigens presentation and its clinical significance. Methods HA117 gene expressions in 36 patients with acute myeloid leukemia (AML) and 14 patients with acute lymphocytic leukemia (ALL),10 patients with idiopathic thromnocytopenic purpura (ITP) were tested by using reverse transcriptase polymerase chain reaction (RT-PCR) technique. Thirty bone marrow specimens of AML patients at different phase were selected, and immunophenotypes were assayed by flow cytometry. ResultsThere was no significant difference ( P > 0.05 ) in HA117 gene expression between ALL (78.57%) and A ML (72.22%). The expressions of HA117 gene in both ALL and AML were significantly higher than in ITP patients ( P < 0.05 ). The HA117 gene was expressed in 75.00% of the initially diagnosed patients and 77.78% of the remission patients and 69.23% of the nonremission patients, all of them had higher expression than the patients with ITP, but there was no significant difference among the three groups. There were 47.06% and 87.88% with HA117 gene expressioned in non-resistance group and resistance group respectively, and the difference between the two groups was significant(P=0.005). HA117 had positively correlation with CD11b(OR =0.59, P=0. 001 ),CD33(OR=0.704, P=0. 000) expression in 30 patients of AML, and had negative correlation with MPO (OR=-0.611, P=0.001). ConclusionHA117 gene overexpression is significantly associated with the clinical resistance. It may also be associated with MDR of acute leukemia. HA117 gene can be used as an important indicator for investigating clinical MDR and predicting prognosis of acute Ieukemia patients. HA1 17 is closely related to CD11b, CD33, MPO expression in AML, and they can be used as biomarkers for evaluating therapy efficacy in AML.
2.Cloning of human csp-B matrix attachment region sequence and construction of its retrovirus vector
Yuxi ZAN ; Li WANG ; Junhe ZHANG ; Tianyun WANG
Chinese Journal of Tissue Engineering Research 2010;14(11):1948-1950
BACKGROUND:Matrix attachment region(MAR)are DNA elements that bound to the nuclear matrices after chromatin digested with restriction endonuclease.Plenty of studies have shown that MAR considered as initiaI point of DNA replication or transcription of regulatory gene.Thereby,construction of MAR expression vector can elevate the overall level of transgene expression,enhance stability of exogenous gene.as welI as increase frequency of stable transfectant cells.OBJECTIVE:To construction pLXSN-CAT recombinant retrovirus vector that containing chloramphenicol acetyltransferase(CAT)via cloning MAR sequence of human.and to explore the influence of MAR on the gene expression.METHODS:An open experiment was performed at the Department of Biochemistry and Molecular Biology.Xinxiang Medical College from September 2007 to December 2007 The PLXSN-CAT vector of CAT was constructed by the laboratory.TaqDNA polymerase,T_4 DNA ligase,DNA Marker,restriction enzyme BamH I,agarose gel DNA purification kit,as well as plasmid purification kit were purchased fromTakara Biotechn0Iogy(Dalian)Co.,Ltd.The sequence of csp-B MAR was amplified by polymerase chain reaction(PCR)method applied to human DNA.The fragment was inserted into retrovirus vector PLXSN-CAT plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.RESULTS AND CONCLUSION:The length of specific fragment applied by PCR was 931 bp,and the recombinant plasmid PLXSN-CAT-MAR presented two bands:5.9 kb and 931 bp using respective restriction enzymes BamH I The sequence of MAR was confirmed by blasting to Genbank(serial numobr:M6271 6).It suggested that MAR had been cloned into PLXSN-CATR vector correctly.The recombinant retrovirus vector PLXSN-MAR was successfully constructed.
3.Regulatory effect of human beta-interferon matrix attachment region on transgene expression in CHO cells
Yuxi ZAN ; Tianyun WANG ; Junhe ZHANG ; Li WANG
Chinese Journal of Tissue Engineering Research 2007;0(29):-
BACKGROUND: Matrix attachment region (MAR), a DNA sequence, is still bound to the nuclear matrices after chromatindigested with restriction endonuclease, not only affects expression of endogenous gene, but also overcames transgenic silence andimproves transcription and expression of exogenous gene. OBJECTIVE: To investigate the influence of ?-interferon MAR of CHO cells on the transgenic expression of chloramphenicolacetyltransferase (CAT). DESIGN, TIME AND SETTING: The opening experiment was performed at the Department of Biochemistry and MolecularBiology, Molecular Institute, Xinxiang Medical College from October 2006 to April 2007. MATERIALS: CHO cell lines were obtained from China Center for Type Culture Collection. The pCATG vector of CAT and G418screening markers were constructed by this laboratory. METHODS: Human ?-interferon MAR by PCR was digested with SacI/KpnI and BamHI/SalI, and was inserted into pCATGvector, which was propagated in Escherichia coli JM109, then extracted and purified followed by enzyme digestion analysis. Vectorof CAT expression cassette and human ?-interferon MAR by the two sides was successfully constructed, and christened aspCAT-MAR. Two methods were compared between CHO cells of pCATG transformation and CHO cells of pCATG-MARtransformation. After G418 selecting, genome DNA of cell lines of G418 was extracted, then primers for PCR to amplify the CATtarget gene fragment was designed. MAIN OUTCOME MEASURES: The activity of CAT was analyzed by ELISA method. It was also tested to see if thepCATG-MAR was stably integrated into genomic DNA in the transfected cells. RESULTS: CHO cells of pCATG transformation was screened to have 16 strains of positive cell, and CHO cells of pCATG-MARtransformation was screened to have 17 strains of positive cell. Human ?-interferon MAR could increase the CAT gene expressionby 2.8 fold. The coefficient of variation of CHO cells of pCATG transformation was 2.065 0, and coefficient of variation of CHOcells of pCATG-MAR transformation was 0.813 1. Genome DNA of stable transformation cell lines was amplified by a fragment of437 bp. The results confirmed the pCAT-MAR vector was stably integrated into genomic DNA. CONCLUSION: Human ?-interferon MAR can increase transgenic expression in CHO cells and decrease the transgenicexpression variation in different transfected cells.
4.Effect of GC-rich DNA fragment on the level of transgene expression in Chinese hamster ovary cells and its position effect
Sijia CHEN ; Chunpeng ZHAO ; Junhe ZHANG ; Xiaoyin WANG ; Tianyun WANG
Journal of Xinxiang Medical College 2017;34(9):794-797
Objective To analyze the effect of GC-rich DNA fragments on the level of transgenic expression in Chinese hamster ovary (CHO) celts and its position effect.Methods The synthetic DNA fragment with GC-rich was cloned into the 5'or 3'or both 5'and 3'ends of expression cassette of expression vector.Three new expression vectors (pIRES-G1,pIRES-G2 and pIRES-G3) which was inserted with the GC-rich DNA fragments in different position were transfected CHO ceils,respectively,and then was observed under fluorescence microscope;the control vector was pIRES-EGFP.Stable transfected cell lines were screened under G418,and enhanced green fluorescent protein(EGFP) expression was analyzed by flow cytometry and the transgenic copy number was detected by quantitative real-time quantitative polymerase chain reaction (qRT-PCR).Results Three expression vectors with a GC-rich DNA fragments in different position were constructed successfully.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector could obviously improve the expression level of vector in CHO cells;and the expression level of the stably transfected CHO cells increased 1.39 fold and 1.32 fold compared to the control vector,respectively;the transgene copy number increased 1.32 fold and 1.24 fold compared with the control vector.While the insertion of GC-rich DNA fragments at 5'end of expression cassette had no obvious effect on the level of gene expression.Conclusion The role of DNA fragment with GC-rich in improving the transgenic expression of CHO cells is related to its position in the vector.The insertion of GC-rich DNA fragments at 3'end and both 5',3'ends of the box of expression vector can improve transgenic expression.
5.Analysis of the therapeutic effect and prognostic factors for carcinoma of parotid gland:a retrospective report of 103 cases
Shaoqing CHEN ; Yu LIU ; Junhe LI ; Yinfang GUO ; Zhe SUN ; Zhenyu YOU ; Xiaolang WANG
Chinese Journal of Clinical Oncology 2014;(12):797-800
Objective: This study aims to analyze the therapeutic effect and prognostic factors of carcinoma of parotid gland (CPG). Methods: Data on 103 CPG patients were retrospectively analyzed. The patients were divided into the simple surgery group (Group One) and post-operative radio-chemotherapy group (Group Two). Kaplan-Meier survival analysis, Log-rank test, and Cox re-gression analysis were employed to analyze the five-year overall survival. Chi-square test was applied to compare the local control rate and recurrence-free survival. Logistic regression analysis was used to determine the correlation between all factors and the local control rate. Results:For all patients, the five-year local control rate, five-year recurrence-free survival rate, and five-year overall survival rate were 71.49%, 69.61%, and 76.10%respectively. The five-year local control ratio (81.96%vs. 61.90%), five-year recurrence-free surviv-al (78.69%vs. 59.52%), and five-year overall survival (88.12%vs. 68.50%) were significantly improved in Group Two compared with Group One. The logistic regression analysis showed that the therapeutic method, T staging, as well as pN(+) neck and tumor differentia-tion were significantly correlated to the five-year local control rate and five-year recurrence-free survival (P<0.01). Cox regression anal-ysis showed that therapeutic method, T stage, as well as pN(+) neck and tumor differentiation were significantly correlated to the five-year overall survival (P<0.01). Conclusion:Post-operative radio-chemotherapy can improve the local control and overall survival rates. This therapeutic method is applicable to patients with T3-4 tumors, as well as pN(+) neck and middle-low differentiation.
6.Townes-Brocks syndrome caused by de novo mutation in spalt-like transcription factor 1 gene: a case report and literature review
Junhe LING ; Yupei WANG ; Jiao TANG ; Xinyuan TIAN ; Duling XU ; Shengju HAO ; Ling HUI
Chinese Journal of Perinatal Medicine 2023;26(7):591-596
Objective:To summarize the clinical features and gene variations in children with Townes-Brocks syndrome (TBS).Methods:The clinical data of a female infant diagnosed with TBS caused by human spalt-like transcription factor 1 ( SALL1) gene mutation in Gansu Maternal and Child Health Hospital in May 2022 were analyzed retrospectively. Relevant articles up to July 2022 were retrieved from several databases including CNKI, VIP, Wanfang, Chinese Medical Journal Network and PubMed with the terms of " SALL1 gene" and "Townes-Brocks syndrome". Patients diagnosed with TBS caused by SALL1 gene mutation were retrieved and the clinical phenotype-genotype correlations in patients with TBS caused by frameshift mutation in SALL1 gene were analyzed and summarized. Descriptive statistical analysis was applied. Results:(1) Clinical data: The index patient was a 40-day-old girl exhibiting major clinical manifestations of polycystic kidney dysplasia, congenital external ear deformity, preaxial polydactyly and recto-perineal fistula. Whole exome sequencing and Sanger sequencing revealed a heterozygous variation of c.420delC (p.S141fs*42) in the SALL1 gene, while the same gene was found to be wild type in her parents and sister. The variant was predicted to be pathogenic (PVS1+PS2+PM2). (2) Literature review retrieved 161 cases of TBS, of which 71 were attributable to a frameshift mutation in SALL1 gene. Clinical phenotypes of the 71 cases and the index case were summarized. TBS was mainly characterized by external ear, hand and anal deformities, sometimes accompanied by hearing loss, abnormal kidney development and foot deformity. A small number of affected cases presented with rare clinical phenotypes such as abnormal eyes, hypothyroidism and abnormal development. At present, the human gene mutation database records 110 variations in the SALL1 gene, with a majority located in exon 2. The most common mutation type was frameshift variation, accounting for 52%, followed by missense variation and nonsense variation. Conclusion:TBS should be considered in children with ear, hand and anal malformations, accompanied by renal dysfunction and hearing loss, and genetic testing is recommended for timely diagnosis.
7.Genetic analysis of a child with Pitt-Hopkins syndrome due to a novel variant of TCF4 gene derived from low percentage maternal mosaicism.
Jiao TANG ; Junhe LING ; Chuan ZHANG ; Shengju HAO ; Jun MA ; Jiaxuan LI ; Lei ZHAO ; Yupei WANG ; Ling HUI
Chinese Journal of Medical Genetics 2023;40(6):680-685
OBJECTIVE:
To explore the genetic etiology of a child with Pitt-Hopkins syndrome.
METHODS:
A child who had presented at the Medical Genetics Center of Gansu Provincial Maternal and Child Health Care Hospital on February 24, 2021 and his parents were selected as the study subjects. Clinical data of the child was collected. Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to trio-whole exome sequencing (trio-WES). Candidate variant was verified by Sanger sequencing. Karyotype analysis was also carried out for the child, and her mother was subjected to ultra-deep sequencing and prenatal diagnosis upon her subsequent pregnancy.
RESULTS:
The clinical manifestations of the proband included facial dysmorphism, Simian crease, and mental retardation. Genetic testing revealed that he has carried a heterozygous c.1762C>T (p.Arg588Cys) variant of the TCF4 gene, for which both parents had a wild-type. The variant was unreported previously and was rated as likely pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (ACMG). Ultra-deep sequencing indicated that the variant has a proportion of 2.63% in the mother, suggesting the presence of low percentage mosaicism. Prenatal diagnosis of amniotic fluid sample suggested that the fetus did not carry the same variant.
CONCLUSION
The heterozygous c.1762C>T variant of the TCF4 gene probably underlay the disease in this child and has derived from the low percentage mosaicism in his mother.
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Transcription Factor 4/genetics*
8.Progress of research on the pharmacological effects and mechanisms of emodin
Congyu WU ; Yue ZHOU ; Luxi SHANGGUAN ; Yani YANG ; Jingya WANG ; Junhe YU ; Shuaishuai GONG ; Junping KOU
Journal of China Pharmaceutical University 2023;54(5):634-643
As an active hydroxyanthraquinone ingredient, emodin is abundant in Chinese medicine herbs, such as Rheum palmatum, Polygonum cuspidatum and Polygonum multiflorum.Modern pharmacological studies have shown that emodin has a variety of pharmacological activities including anti-tumor, anti-inflammatory and immunoregulatory, antibacterial and anti-viral effects, myocardial protection, neuroprotection, renal protection, bone protection, antifibrosis and so on, which indicate its high medicinal value and broad application prospects.This article aims to summarize the progress in the pharmacological activity and mechanism of action of emodin published in domestic and international journals over the last 5 years and highlight the potential targets and molecular signaling pathways linked with emodin, so as to provide some clues and references for further development and clinical application of emodin.