1.miRNA-199 a-5 p inhibit the invasion of MDA-MB-231 cells via regulating ERK5 through SP1
Limin ZHAI ; Shuo YANG ; Wentong LI
Chinese Journal of Clinical and Experimental Pathology 2015;(9):981-985
Purpose To study the effect and mechanism of miR-199a-5p on the invasion of breast cancer MDA-MB-231 cells. Meth-ods miR-199a-5p mimic was transfected into MDA-MB-231 cells. Influence of miR-199a-5p on the invasion of MDA-MB-231 cell was displayed by Transwell, the expression of epithelial-mesenchymal transition ( EMT) molecular markers ( E-cadherin, vimentin) regulated by miR-199a-5p was determined using immunofluorescence and Western blot. Western blot was employed to assess the levels of ERK5, pERK5, EGF and SP1 in MDA-MB-231 cells dealt with miR-199a-5p mimic and LNA-siRNA. Chromatin immunoprecipita-tion (CHIP) was applied for displaying the reaction of SP1 with ERK5 promoter. Results miR-199a-5p could inhibit the invasion of MDA-MB-231 cells, decrease the expression of vimentin and enhance E-cadherin. Meanwhile, miR-199a-5p decreased the expression of ERK5 and pERK5, the levels of EGF and SP1 were also downregulated. On the contrary, the levels of EGF, SP1, ERK5 and pERK5 were enhanced by employing LNA-siRNA targeting miR-199a-5p. SP1 could bind with ERK5 promoter. Conclusions miR-199a-5p could reduce the expression of ERK5 and pERK5 through regulating EGF and SP1, which functioning the inhibitory effect on invasion of MDA-MB-231 breast cancer cells.
2.Simultaneous determination of 7 kinds of bioactive components of Ligusticum chuanxiong Hort. by HPLC-MS
Chinese Traditional Patent Medicine 1992;0(08):-
AIM: To establish an HPLC-MS for determining seven bioactive components,including vanillin ferulaic acid,ligustrazine,senkyunolid I,H,and A,and ligustilide in Ligusticum chuanxiong Hort. . METHODS: The bioactive components were extracted by ultrasonic method,and then were separated by reverse HPLC,and finally were determined by the multiple reaction mode of MS. RESULTS: The detected limits of the seven bioactive components such as vanillin were 40,20,80,20,10,60 and 50 ng/mL,respectively. Their linear range were 1450,4-490,1-625,2-360,2-700,1-244 and 21-210 ?g/mL,respectively. The average contents of the above components in Ligusticum chuanxiong Hort. were 0. 12,0. 25,0. 41,0. 99,0. 30,8. 96 and 11. 25 mg/g,respectively. CONCLUSION: The proposed method has the characteristic of simplicity,rapidty and high sensitivity. It could be used in analyzing the whole components of Ligusticum chuanxiong Hort. .
3.The hospital scientific research evaluation and influence factors analysis
Limin PAN ; Junxia ZHAI ; Biao XI
Chinese Journal of Medical Science Research Management 2014;27(3):253-256,266
Scientific research to a great extent,represents the comprehensive strength of a unit,directly determine the development prospects of this unit.At present,our country many medical institutions of scientific research strength is low,potential is insufficient,can't meet the development of medical institutions.This study from the aspects of management performance,to evaluate scientific research ability from various perspectives,statistical analysis of its influencing factors at the same time,Reflects the technical level of this unit,discipline construction,scientific research and development and performance,produced by scientific management for the unit provides an objective basis for scientific research management departments to formulate relevant policies to explore to improve the new management starting point,thus achieve the goal of science and technology.
4.Sensorineural hearing impairment resulted from radiochemotherapy in patients with nasopharyngeal carcinom
Yumei WEI ; Baosheng LI ; Anting XU ; Limin ZHAI ; Haiqun LIN
Chinese Journal of Radiation Oncology 2009;18(2):93-95
Objective To compare the severity of sensorineural hearing impairment resulted from radiotherapy (RT) and radiochemotberapy (CRT) in patients with nasopharyngeal carcinoma (NPC). Methods Between March 2002 and May 2007, 100 initially diagnosed NPC patients in Shandong Tumor Hospital and Qi Lu Hospital were randomized to RT group and CRT group. All patients underwent intensity modulated radiation therapy. In CRT group, concurrent and adjuvant CDDP were administered (CDDP 25 mg/m2/d for 3 days to 4 cycles). Pure tone auditory threshold examination was performed 1 week ,6 months, 1 year and 2 years after the completion of radiotherapy. Statistical analyses were performed using Mann-Whit-ney U test,chi-square test and Fisber's exact probability test. Results The high-frequency threshold was significantly increased in CRT group comparing with RT group at 1- and 2-year after the treatment. In RT group, the hearing threshold was impaired immediately after the treatment, partially recovered within the first year but impaired again after 2 years. In CRT group, hearing threshold was impaired at the same time and kept getting worse until 1 year after radiotherapy, which could not be recovered. Conclusions Patients with NPC treated with radiotherapy and concurrent/adjuvant chemotherapy have more severe sensorineural hearing impairment comparing with those with radiotherapy alone, especially to the high frequency sound in the speech range. Inner ear tissue tolerance should be redefined for patients receiving radiochemotherapy.
5.Current situation of scientific research administrator in Primary Hospital and training Countermeasures
Limin PAN ; Xiantao HUANG ; Kuanlei WANG ; Chao JING ; Junxia ZHAI
Chinese Journal of Medical Science Research Management 2014;27(5):568-570
With the advent of the knowledge economy,hospital management of scientific research work is facing new challenges.At present,administrators in charge of scientific research in most primary hospitals of China do not have the systematic training on knowledge and skills involving research management.Thus,training a master business and familiar with the operation of modern hospital science and technology management of high-quality management personnel is urgent needed.As long as Do a good job on training of the management personnel of scientific research in the primary hospital,can enrich the scientific research management team,do good on the management work of sci ence and technology in the hospital,hospital work to be continuous development of science and technology and enhance.
6.Effects of metformin on expression of renal tissue AGEs in type 2 diabetic rats
Yeqiong LI ; Shandong YE ; Limin ZHAI ; Wen HU
Chinese Pharmacological Bulletin 2016;32(5):703-706,707
Aims To observe the influences of met-formin ( MET ) on the expression of renal tissue ad-vanced glyclation end-products( AGEs) protein and its receptor mRNA ( RAGE mRNA ) in type 2 diabetes mellitus (T2DM) model rats, and to discuss the mech-anism of the MET in the treatment of diabetic nephrop-athy ( DN ) . Methods The rat model of T2 DM was established by fed with high-fat diet and intraperitoneal injection of low-dose of streptozotocin ( STZ ) . All rats were randomly divided into metformin group( MET,300 mg·kg-1 ·d-1 ) , glyburide group( GLY,5 mg·kg-1 ·d-1),T2DM model group(T2DM) and normal con-trol group ( NC ) . After 8 weeks ’ observation, blood glucose ( BG ) , glycated hemoglobin ( HbA1 c ) , blood urea nitrogen(BUN), urinary albumin,urinary AGEs and urine creatinine were detected. The expression of renal tissue AGEs was detected by immunohistochemis-try assay, and the expression of RAGE mRNA was measured by real-time PCR. Results The levels of BG, HbA1c , urinary albumin/urine creatinine ( UACR ) , glomerular basement membrane thickness ( GBMT ) in MET group and GLY group were signifi-cantly lower than those of T2DM group, while higher than those of NC group(P<0. 05), the levels of BG, FINS and HbA1 c were not statistically significant be-tween MET and GLY group ( P >0. 05 ) . The urinary AGEs/urine creatinine( UGCR) , the expressions of re-nal tissue AGEs and RAGE mRNA in MET group and GLY group were significantly decreased compared with those of T2 DM group ( P < 0. 05 ) , but higher than those of NC group ( P <0. 05 ) . The UGCR, the ex-pressions of AGEs and RAGE mRNA in MET group were lower than those of GLY group(P<0. 05). Con-clusion MET can reduce the accumulation of AGEs in the renal tissue,and down-regulate the over-expres-sion of RAGE mRNA in T2DM rats.
7.Effects of Chinese herbal medicines Shengmai injection and Xuesaitong injection on ventricular fibrillation threshold and connexin 43 expression in rats with myocardial infarction.
Aiming WU ; Dongmei ZHANG ; Lixia LOU ; Jianying ZHAI ; Xiying Lü ; Limin CHAI ; Shuoren WANG
Journal of Integrative Medicine 2011;9(7):775-82
To explore the effects of Shengmai injection and Xuesaitong injection, compound Chinese herbal medicines for replenishing qi and activating blood, on ventricular fibrillation threshold, heart structure and connexin 43 (Cx43) expression in rats with myocardial infarction (MI).
8.Expression and correlation of PKCζ, MMP-2 and MMP-9 in breast cancer
Shuo YANG ; Hongli LI ; Wentong LI ; Lu YANG ; Ming NI ; Limin ZHAI ; Chonggao YIN ; Baogang ZHANG
Chinese Journal of Clinical and Experimental Pathology 2014;(9):958-962
Purpose To investigate the expressions of PKCζ, MMP-2, and MMP-9 in breast cancer and the relationship with the inva-sion and metastasis of breast cancer. Methods The expression of PKCζ, MMP-2 and MMP-9 in 100 cases with breast cancer was as-sessed with immunohistochemistry PV 9000 method. PKCζ-siRNA was transfected into MDA-MB-231 cell lines, called siPKCζ/MDA-MB-231. While siRNA construct containing a scrambled sequence was transfected into MDA-MB-231 cells to generate control cells, which were designated as Scr/MDA231 cells. Western blotting was used to measure the expression of PKCζ in transfected cells, and the Transwell invasion assay was used to detect the invasion ability in vitro. The content of MMP-2, MMP-9 were measured by ELISA. Results The expression rates of PKCζ, MMP-2 and MMP-9 in breast cancer tissues were 62.5%, 37.5% and 32.5%, and there were significant differences among them (P<0.05). The expression of PKCζwas much higher than those in the normal breast tissues nearby. The expression of PKC protein was assoiated with lymph node metastasis, distant metastasis (P<0.01), but was not correla-ted with other clinicopathologic parameters, such as age, tumor size, histological type, ER, PR, and so on (P>0.05). The expres-sion of PKCζ, MMP-2 and MMP-9 were lower in siPKCζ/ MDA-MB-231 group than those in scr/ MDA-MB-231 group, and the in vitro invasion ability was significantly decreased (P<0.05). Conclusions PKCζ can promote the invasion and metastasis of breast canc-er, and correlated with the expression of MMP-2, MMP-9(P<0.05).
9.Noninvasive prenatal screen of trisomy-21 using maternal plasma fetal free RNA allelic ratio
Yajuan XU ; Shanshan ZHAI ; Xiaohua LUO ; Yingying ZHANG ; Limin RAN ; Lidan REN
Chinese Journal of Obstetrics and Gynecology 2015;(8):568-575
Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.
10.Efficacy of intensity-modulated radiotherapy combined with chemotherapy for nasopharyngeal carcinoma in children and adolescents
Dongqing WANG ; Xiujuan CAO ; Wei DONG ; Shui YU ; Xinhua YANG ; Man HU ; Limin ZHAI
Chinese Journal of Radiological Medicine and Protection 2017;37(2):125-131
Objective To evaluate the treatment outcomes and toxicities of intensity-modulated radiation therapy (IMRT) combined with chemotherapy for children and adolescents with nasopharyngeal carcinoma.Methods Forty-three nasopharyngeal carcinoma patients less than 19 years old were recruited between April 2010 and April 2016.All patients were treated with IMRT (total dose 61.2-76 Gy) combined with cisplatin based chemotherapy.The Kaplan-Meier test was used to calculate overall survival (OS) and progression-free survival (PFS).The patient's clinical characteristics,side effects and longterm effects of treatment were retrospectively analyzed.Results Among 43 patients,there were 29 (67.4%) male and 14 (32.6%) female,and the median age was 14 years old (range,6-18 years).According to AJCC 7thstaging system,2 patients were in stage Ⅱ,26 in stage Ⅲ,7 in stage ⅣA and 8 in stage Ⅳ B.All patients were confirmed as non-keratinizing carcinoma.The positive rates of EB virus VCAIgA was 53.8% (7/13),and Rta-IgG was 60.0% (6/10) before treatment.The median radiation dose was 70 Gy (range,61.2-76 Gy) to the primary tumor.Thirty-three (76.7%) patients received neoadjuvant chemotherapy,with 20 (46.5%) and 36 (83.7%) patients treated by concurrent and adjuvant chemotherapy,respectively.With a median follow-up of 24 months (range,3-76 months),the 5-year OS and PFS ratios were 75.3% and 64.7%,respectively.There were 5 patients (11.6%) occurred to bone metastasis within 2 years after treatment.Hypothyroidism was reported in 47.4% (9/19) patients after IMRT.Conclusions Nasopharyngeal carcinoma in childhood and adolescence is mostly locally advanced diseases with poor differentiation.IMRT combined with chemotherapy produce a well treatment outcome with good tolerance in children and adolescents patients.The most common treatment failure bone metastasis.Radiation-induced hypothyroidism is common.