1.Expression of recombinant plasmid of homo heterogeneous nuclear ribonucleoprotein E1 in SH-SY5Y cells
Lirong HUO ; Lanying WANG ; Junhua ZOU ; Nan ZHONG
Chinese Journal of Tissue Engineering Research 2014;(42):6769-6774
BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.
2.Characteristics of chromosome karyotype and marker chromosome of human breast cancer cell lines Bcap-37 and MCF-7
Junhua ZOU ; Junzheng YANG ; Lingfeng MIN ; Jiaying LIU
Chinese Journal of Tissue Engineering Research 2005;9(18):258-259
BACKGROUND: Cytogenetic evidence suggests that chromosomal alteration are not randomly occurring events and some malignancies are characterized by specific chromosome abnormalities, which provides cytogenetic basis for the expression of oncogenes.OBJECTIVE: To investigate the characteristics of chromosomal karyotype and marker chromosome of breast cancer cell lines Bcap-37and MCF-7 by means of G-banding chromosomal analysis.DESIGN: A controlled experiment with breast cancer cells as observation subjects.SETTING: Department of Medical Genetics, Peking University Health Science Center.MATERIALS: This study was carried out in the Department of Medical Genetics of Peking University Health Science Center from April 1991 to May 1992 using breast cancer cell lines MCF-7 and Bcap-37.METHODS: The chromosomes of human breast cancer cell lines Bcap-37and MCF-7 were obtained by growth synchronization induced by hypothermia and colchicines treatment. The cells at prometaphase or metaphase underwent G-banding chromosomal analysis. For each cell line, 50 to 60 mitotic figures were counted and 15 or 16 G-binding karyotypes were analyzed, including the mitotic figure at the level of about 320- and 500-band .MAIN OUTCOME MEASURES:Abnormality in chromosome number and structural aberration of the two breast cancer cell lines.RESULTS:The modal chromosomal number of Bcap-37 cell line was 63, of which 17 marker chromosomes had identifiable structure, as compared with 13 out of 56 chromosomes in modal number of MCF-7 cell line.CONCLUSION: Both of the two breast cancer cell lines have complex cytogenetic abnormality in the modal number and structure of the chromosomes, which might result in the rearrangement of DNA sequence of the cancer-related genes or DNA depletion, so as to play important roles in the pathogenesis and development of breast cancer.
3.Effect of electroacupuncture at acupoint Guanyuan on excitability of detrusor and expression of BDNF and TrkB in spinal cord of rat with urinary retention after spinal cord injury
Junhua WANG ; Bangguo CHEN ; Jing YIN ; Weigeng ZOU ; Xiaojuan LUO
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(4):232-235
Objective To study the effect and mechanism of electroaeupuncture at acupoint Guanyuan on expression of brain-derived neurotrophic factor (BDNF) and tropomyosine receptor kinase B (TrkB) in spinal cord and excitability of detrusor of rat with urinary retention after spinal cord injury. Methods The model of urinary retention after spinal cord injury was established by using weight drop method. Eighty female Sprague-Dawley rats were randomly divided into four groups, namely, a normal, a sham, a model and an electroacupuncture (EA) group, with 20 rats in each group. The EA group was given electroacupuncture at aeupoint Guanyuau once a day for 10 days in total, while the other groups was given no treatment. After 10 days, the expression level of BDNF and TrkB in spinal cord of injury site was measured by immunohistochemistry staining, and excitability of detrusor was measured by using the muscle strips in vitro. Results It was revealed that, with regard to the expression level of BDNF and TrkB in spinal cord,the model group exceeded the normal and sham group, the EA group significantly exceeded the other groups ( P < 0. 05 ). With regard to the excitability of detrusor, the model group was significant-ly lower than the normal and sham group. The EA group significantly exceeded the model group ( P < 0.05 ). Conclusion Electroacupuncture at acupoint Guanyuan can raise the expression level of BDNF and TrkB in spinal cord and increase the excitability of detrusor in rats with urinary retention after spinal cord injury.
4.A new triterpenoid saponin julibroside J_(24) from the stem bark of Albiza julibrissin
Kun ZOU ; Bin WANG ; Yuying ZHAO ; Junhua ZHENG ; Ruyi ZHANG ;
Journal of Peking University(Health Sciences) 2004;0(01):-
Objective:To separate chemical and active constituents from the stem bark of Albiza julibrissin . Methods:Chromatography and spectral methods were used. Results:3 O ? D xylopyranosyl (1→2) ? L arabinopyranosyl (1→6) ? D glucopyranosyl 21 O {(6 S) 2 trans 2 hydroxymethyl 6 methyl 6 O [3 O (6 S ) 2 trans 2 hydroxymethyl 6 methyl 6 hydroxy 2,7 octadienoyl) ? D xylopyranosyl] 2,7 octadienoyl} acacic acid 28 O ? D glucopyranosyl (1→3) [? L arabinofuranosyl (1→4)] ? L rhamnopyranosyl (1→2) ? D glucopyranosyl ester (1) was separated and identified. Conclusion: Compound 1 is a new saponin named as Julibroside J 24 .
5.Results analysis of serum alanine transaminase measurement in 23 clinical laboratories with target value assigned by reference method
Weimin ZOU ; Junhua ZHUANG ; Xianzhang HUANG ; Zengwen LIN ; Yun LI ; Jianbin WANG ; Jiaqi LI
Chinese Journal of Laboratory Medicine 2015;(5):318-320
Objective To explore the feasibility of inter-laboratory comparison and trueness evaluation among clinical laboratories, and assess the quality of participants′measurement, by measuring the activity of alanine aminotransferase ( ALT ) in patient serum samples.Methods Method comparison study was used.Five patients serum samples, whose target values were assigned by two international candidate reference laboratory with reference method of ALT without pyridoxal phosphate, were measured by 23 routine laboratories.The bias between measurement result of each participant and the mean of reference laboratories was calculated, and then compared to allowable bias 6%.Calculate the mean value and the relative bias.Results Compared with the mean of reference laboratories, the maximum absolute value of bias among the 23 routine laboratories was 31.27%.The rate range which bias was less than the allowable bias was 26.09%-73.91 %.The bias acceptability of 8 participants were more than or equal to 80%;15 participants were less than or equal to 60%; and 3 participants were 0%.Conclusions Using patient serum samples and values assigned by reference method is an effective way to carry out inter-laboratory comparison and trueness evaluation.It can reflect the quality of measurement more truly.
6.EGFR gene mutation status, ERCC1 and Ki-67 protein expression in non-small cell lung cancer and relationship with clinicopathologic features
Yiming HAN ; Jie ZHENG ; Yunhui JIANG ; Jinhua SHEN ; Lan RAO ; Wanting FAN ; Junhua XIE ; Xianjin ZOU
Chinese Journal of Clinical and Experimental Pathology 2015;(7):759-763
Purpose To study the status of EGFR mutations and the expression of excision repair cross-complementation group 1 ( ER-CC1) and Ki-67 protein in patients with non-small cell lung cancer (NSCLC) and to examine the relationship between their expression and clinicopathologic features. Methods EGFR mutations were analyzed with DNA sequencing, and the expression of ERCC1 and Ki-67 protein was examined by immunohistochemistry EnVision. The relationship of EGFR mutations with the expression of ERCC1and Ki-67 and the clinicopathological features were analyzed. Results EGFR mutations were detected in 143 (143/291, 49. 1%) of the 291 specimens. EGFR mutations were found more frequently in women, non-smokers and adenocarcinoma. The difference of EGFR muta-tion rate between the histological subtypes according to the IASLC/ATS/ERS classification of lung adenocarcinoma was significantly ( P=0. 008). The mean tumor diameter was smaller in patients with EGFR mutations than in those with wild-type EGFR (P=0. 020). EGFR mutations were not related to age, lymph node metastasis. However, EGFR mutations were not related to the expression of ER-CC1 and Ki-67 protein (P>0. 050). Conclusions EGFR mutation is closely linked to several clinicopathological factors, such as gender, differentiation, and histological subtype. There is heterogeneity of EGFR mutation in patients with NSCLC. EGFR mutations were not related to the expression of ERCC1 and Ki-67 protein.
7.Procollagen type 1 N-terminal propeptide and beta-collagen special sequence for the diagnosis and treatment of osteoporosis
Jun ZOU ; Chenxi YUAN ; Hongjun ZHU ; Ying LU ; Min LIN ; Yijia CHEN ; Junhua WANG ; Jiaxuan XU
Chinese Journal of Tissue Engineering Research 2015;(29):4731-4735
BACKGROUND:Procolagen type 1 N-terminal propeptide (P1NP) and β-colagen special sequence(β-CrossLaps) are two bone metabolic markers that are closely related to osteoporosis. Combined detection of bone metabolic markers and bone mineral density is of clinical significance for the diagnosis of osteoporosis. Bone metabolic markers are ideal indicators to predict fractures, which can compensate for the lack of bone density test. OBJECTIVE:To introduce the application of bone metabolic markers in the monitoring of drug efficacy on the treatment of osteoporosis as wel as in the prediction of fracture risks in recent 20 years and to explore the clinical values of P1NP and β-CrossLaps to assess the therapeutic efficacy on osteoporosis and risks for osteoporotic fractures. METHODS:A computer-based search of CNKI and SCI databases were performed for relevant articles published from 2000 to 2014 using the keywords of “serum bone metabolic markers; osteoporosis; bone mineral density” in Chinese and English, respectively. Finaly, 44 articles meeting the inclusive criteria were reviewed. RESULTS AND CONCLUSION:This paper analyzes the source and detection mechanisms of P1NPand β-CrossLaps and then compares their advantages in the therapeutic effect assessment of osteoporosis. Serum bone metabolic markers cannot only reflect the dynamic changes of bone metabolism, but also have earlier changes than the bone mineral density. Both P1NPand β-CrossLaps are very important for assessing the early diagnosis of osteoporosis as wel as anti-osteoporosis drug efficacy.
8.Identification of novel partner proteins of PCBP1
Lirong HUO ; Chen SHEN ; Weina JU ; Junhua ZOU ; Wu YAN ; W.ted BROWN ; Nanbert ZHONG
Journal of Peking University(Health Sciences) 2009;41(4):402-408
Objective: PCBP1 is a family member of heterogeneous nuclear ribonucleoproteins (hnRNPs) that belong to RNA-binding proteins and bear three KH domains. The protein plays a pivotal role in post-transcriptional regulation for RNA metabolism and RNA function in gene expression. We hy-pothesized and were going to identify that the regulatory function of PCBP1 is performed through different complexes of proteins that include PCBP1. Methods: To test our hypothesis, approaches of protein wal-king with a yeast two-hybrid system (Y2H), pulling down in yeasts, co-immunoprecipitation and immu-nofluorescent microscopy assay were employed in this study. The PCBP1 was used as the initial "walker" to search for its interaction partner(s). Results: Candidate proteins including MYL6, PECAM1, CSH1,RAB7, p57KIP2, ACTG1, RBMS1 and PSG4-1ike were identified with selection mediums and preceding methods. Conclusion: With these candidate protein molecules, some protein complexes associating with PCBP1 are proposed, which may help in a better understanding of physiological functions of PCBP1 and proved evidence that PCBP1 is involved in variant biological pathways.
9.The effect of electro-acupuncture on bladder detrusor apoptosis in rats with urinary retention after spinal cord injury
Junhua WANG ; Bangguo CHEN ; Jing YIN ; Weigeng ZOU ; Xiaojuan LUO ; Xiaoming ZHANG
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(6):380-384
Objective To study the effect of electro-acupuncture(EA)on bladder detrusor apoptosis in rats with urinary retention after spinal cord injury(SCI),compare the effectiveness of different acupoints and explore possible mechanisms.Methods One hundred female adult Sprague-Dawley rats were randomly divided into 5 groups:a normal group,a sham operation group,a model control group,a Guanyuan acupoint(RN4)EA group and a Shuidao acupoint(ST28)EA group,with 20 rats in each group.A model of urinary retention after SCI was established using the weight drop method.The EA groups were given EA once daily for 10 d.The other groups had no treatment.After 10 d of EA,an electron microscope was used to observe the bladder detrusor ultrastructure,and the terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) method and immunohistochemistry were used to observe any apoptosis in the detrusor smooth muscle cells and the expression of apoptosis-related genes bcl-2 and bax.Results After EA treatment,expansion of the rough endoplasmic reticulum was reduced significantly,and the number of edemic mitochondria was reduced compared with the model control group.In the normal and sham operation groups,little apoptosis was observed,and there was no significant difference between the two groups.Apoptosis indices(AIs)were higher in the model control group than in the normal group.The average AI in the Guanyuan EA group was lower than that in the Shuidao group.In the Guanyuan group,bax protein expression was less than that in the Shuidao group and the model group.In the Guanyuan group,bcl-2 protein expression was not significantly different from that in the Shuidao group and the model control group.Conclusion EA at the Guanyuan or Shuidao aeupoint can restrain bladder detrusor apoptosis in rats with urinary retention after SCI.EA may improve the ultrastructure of the detrosor urinae, reduce the expression of bax, change the ratio of bcl-2 to bax, and thus decrease apoptosis.The effect of EA at the Guanyuan acupoint is superior to that at the Shuidao acupoint.
10.Detection of serum IFN-?, IFN-?, NO and iNOS in patients with SARS and its clinical significance
Xinzhong WU ; Junhua ZHUANG ; Lin CHEN ; Xianzhang HUANG ; Lin LIN ; Guoli ZOU
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To study the serum level of interferon ( IFN)-?, IFN-?, nitric oxide (NO) and nitric oxied synthase (iNOS) in severe ac ute respiratory syndrome (SARS) patients and to expl ore its significance. METHODS: IFN-?, IFN-?, NO and iNOS were determined in SARS patients during the early, recovery and follow-up stage, the health doctors and nurses and health people, respectively. RESULTS : The serum level of IFN-? during the early stage was higher than that of other groups (P