1.Expression of miR-634 in hepatocellular carcinoma and its effect on biological behavior of Hepatocellular carcinoma cells
Fei LI ; Dexin LI ; Guangpeng ZHOU
Chinese Journal of Immunology 2016;32(8):1160-1164
Objective:To detect the expression level of microRNA-634 (miR-634) in hepatocellular carcinoma (HCC) and its regulatory effect on the common biological behavior of hepatocellular carcinoma cells .Methods: Real-time fluorescence quantitative PCR (RT qPCR) method was used to detect HCC cell lines (HepG2, SMMC7721, BEL7402, bel7404, SNU739), 69 cases of HCC tissues and matching relative quantification of miR-634 paracancerous tissues and analysis of relationship between miR-634 expression and HCC patients gender, age, tumor size, degree of differentiation, child Pugh classification, BCLC staging, portal vein tumor thrombus and liver metastasis , while building a miR-634 eukaryotic expression vector and transfected into hepatocellular carcinoma cell lines, using live cell counting kit-8 CCK-8, flow cytometric annexin V/PI double staining and Transwell experiment to detect the transfection miR-634 on cell proliferation , apoptosis and invasion effects .Results:Compared with the normal human liver cell line L-02 and hepatocellular carcinoma cells miR-634 were decreased ( P<0.05 ) , the expression followed by HepG 2>SNU739>Bel7402>Bel7404>SMMC7721;69 cases of hepatocellular carcinoma ( HCC ) of miR-634 level ( 0.253 ±0.019 ) and lower than that of the matched paracancerous tissues ( P<0.05 ) , and related with the tumor size , degree of differentiation , BCLC stage , portal vein tumor thrombus and liver metastasis ( P<0.05 ) .Over expression in the transfected group 24-96 h after miR-634 level continues to rise , control group and blank vector transfected group differences were statistically significant ( P<0.05 );and the control transfection group and blank group compared to transfection proliferation inhibition rate , apoptosis rate was increased , but wear the number of cell membrane decreased, the difference was statistically significant (P<0.05).Conclusion:miR-634 in hepatocellular carcinoma tissues and cells were low expression and related to clinical pathological parameters , raised its level can inhibit the proliferation and invasion of hepatocellular carcinoma cells and induce apoptosis , for the prevention and treatment of liver cancer has an important reference value .
3.Intratympanic Dexamethasone in the Treatment of Acute Low-Frequency Hearing Loss
Jie FEI ; Weidong ZHOU ; Jihong LI
Journal of Audiology and Speech Pathology 2010;18(2):150-152
Objective To explore the efficacy of intratympanic dexamethasone injection in treating acute low-frequency hearing loss(ALHL).Methods Thirty-seven ALHL cases not responsive to intravenous dexamethasone and vasodilator medications were enrolled in the study and divided into two groups.In the treatment group,dexamethasone was injected intratympanically once per day to 18 cases for a seven-day history with unilateral ALHL and 1 case with bilateral ALHL.In the control group,18 cases with unilateral ALHL were given intravenous vasodilator medications for 7 days.All patients were followed up for 1~ 2 years.Results In the treatment group with unilateral ALHL,8 cases achieved complete recovery,7 cases achieved partial recovery and no change was observed in the other 3 cases.The efficiency was 83.3 %.2 cases showed recurrent low-frequency hearing loss during the follow-up period.One case with bilateral ALHL did not respond to the treatment and no recurrence was observed.No side effect was observed in the study.In the control group,3 cases completely recovered,5 cases partially recovered and 10 cases showed no improvement.The efficiency was 44.4%.5 cases recurred and one of them progressed to Meniere's disease.There was a statistically significant difference in efficiency of patients with unilateral ALHL between two groups(P<0.05).Conclusion Intratympanic dexamethasone application was an effective and safe treatment option for ALHL.
4.Pattern of Explicit Self-enhancement of Chinese Undergraduate
Fei TENG ; Dengfeng WANG ; Li ZHOU
Chinese Journal of Clinical Psychology 1993;0(01):-
Objective: Self-enhancement is a kind of motive that urges people to attend exclusively to the positive aspects of himself,it is so intense that individuals even tend to distort the threatening information to some degree so that they can maintain a positive self-view. This research is to investigate the pattern of self-enhancement of Chinese people. Methods: Self-enhancement was measured by requiring participants to make self attribution as well as other attribution of their success and failure on eight factors. Results: There existed strong self-enhancement motive in Chinese individuals. Specifically,Chinese undergraduates attribute their success to inner factors such as personality and effort. This trends also evident in the conditions in which participants were asked to speculate the attribute pattern of other people like his/her mother on their own success and failure. Conclusion: Chinese undergraduates tend to enhance the self-image in other people's eyes.
5.THE CHANGE IN mGluR1? IN CA_1 AFTER INFRASONIC DAMAGE AND THE EFFECT OF ITS ANTAGONIST MCPG
Zhigang LI ; Zhou FEI ; Xian ZHANG
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
To study the change in mGluR1? in CA 1 after infrasonic damage and the effect of its antagonist MCPG,120 SD rats were randomizied into infrasonic damage group and MCPG therapy group.The two groups were subdivided into control group and 1 challenge, 7 chellenges, 14 challenge groups respectively,each group consisted of 15 rats. Rats were exposed to 8Hz 130dB infrasound as designed,2h for each challenge.The mGluR1? was stained by immunohistochemistry method.The changes in mGluR1? and cell pattern were observed under light and electronic microscopes.The results showed the number of mGluR1? positive neurons increased after 1 challenge infrasonic damage( P 0 05).MCPG had an effect to protect neuron from infrasonic damage as identified by morphology.It is concluded that the change of mGluR1? activity could mediate excitotoxicity and was one of the major factors related with neuron injury after infrasound.
6.Identification of Candida albicans genes with cDNA technique from LongSAGE tags
Yongping LI ; Cunjian ZHOU ; Fei HAO
Journal of Third Military Medical University 2003;0(20):-
Objective To identify the LongSAGE tags from yeast LongSAGE library and hyphal LongSAGE library of Candida albicans. Methods The technique called the generation of longer cDNA fragments from serial analysis of gene expression (SAGE) tags was used for gene identification (GLGI). The SAGE tags of 17 bases were converted into their corresponding 3′ cDNA fragments covering hundred bases, and the sequence was analyzed. Results Fifteen out of twenty tags were stably and efficiently extended with the base size between 200-1 000 bp. The sequence was considered to represent a known gene since it matched a full-length transcript sequence with over 95% similarity in the same orientation, including the same 17-bp SAGE tag sequence. Conclusion By using novel SAGE tags as primer, we can identify efficiently many novel transcripts. A combined application of SAGE-GLGI can be used to define the boundary of expressed genes in the genomic sequences in Candida albicans and in other eukaryotic genomes.
7.Wudang cherry ameliorates urate underexcretion and renal dysfunction in hyperuricemic mice
Chinese Journal of Pharmacology and Toxicology 2017;31(10):991-991
OBJECTIVE To investigate effects of Wudang cherry on urate excretion and renal function and examined whether renal organic ion transporters were involved in potassium oxonate-induced hyperuricemic mice. METHODS The model of hyperuricemic mice was induced by intraperitoneal injection of potassium oxonate (250 mg·kg- 1) for 7 d. Water extracts of Wudang cherry at 500 mg·kg- 1 were orally administered to hyperuricemic mice for 7 d, benzbromarone (20 mg·kg- 1) and allopurinol (20 mg · kg- 1) were given as positive controls, vehicle control group was given equal normal saline. Serum and urine levels of uric acid were measured in hyperuricemic and normal mice. Simultaneously, the mRNA and protein levels of mouse urate transporter 1 (mURAT1), glucose transporter 9 (mGLUT9), organic anion transporters (mOAT1 and mOAT3), ATP- binding cassette, subfamily G, membrane 2 (mABCG2) and organic cation/carnitine transporters (mOCT1, mOCT2, mOCTN1 and mOCTN2) in the kidney were analyzed by Western blot, RT-PCR, immunohistochemical and immuno?fluorescent assay, respectively. RESULTS Wudang cherry significantly reduced serum uric acid levels and increased urine uric acid levels in hyperuricemic mice. And it effectively reversed potassium oxonate-induced alterations in renal mURAT1, mGLUT9, mOAT1, mOAT3 and mABCG2 mRNA and protein levels, resulting in the enhancement of renal urate excretion in mice. Moreover, Wudang Cherry increased renal mOCT1, mOCT2, mOCTN1 and mOCTN2 mRNA and protein levels, and improved renal impairment in this model. CONCLUSION Wudang cherry processes uricosuric and nephroprotective actions by regulating renal organic ion transporters in hyperuricemic mice.
8.Wudang cherry ameliorates urate underexcretion and renal dysfunction in hyperuricemic mice
Chinese Journal of Pharmacology and Toxicology 2017;31(10):1010-1011
OBJECTIVE To investigate effects of Wudang cherry on urate excretion and renal function and examined whether renal organic ion transporters were involved in potassium oxonate-induced hyperuricemic mice. METHODS The model of hyperuricemic mice was induced by intraperitoneal injection of potassium oxonate (250 mg·kg- 1) for 7 d. Water extracts of Wudang cherry at 500 mg·kg- 1 were orally administered to hyperuricemic mice for 7 d, benzbromarone (20 mg·kg- 1) and allopurinol (20 mg · kg- 1) were given as positive controls, vehicle control group was given equal normal saline.Serum and urine levels of uric acid were measured in hyperuricemic and normal mice. Simultaneously, the mRNA and protein levels of mouse urate transporter 1 (mURAT1), glucose transporter 9 (mGLUT9), organic anion transporters (mOAT1 and mOAT3), ATP- binding cassette, subfamily G, membrane 2 (mABCG2) and organic cation/carnitine transporters (mOCT1, mOCT2, mOCTN1 and mOCTN2) in the kidney were analyzed by Western blot, RT-PCR, immunohistochemical and immuno?fluorescent assay, respectively. RESULTS Wudang cherry significantly reduced serum uric acid levels and increased urine uric acid levels in hyperuricemic mice. And it effectively reversed potassium oxonate-induced alterations in renal mURAT1, mGLUT9, mOAT1, mOAT3 and mABCG2 mRNA and protein levels, resulting in the enhancement of renal urate excretion in mice. Moreover, Wudang cherry increased renal mOCT1, mOCT2, mOCTN1 and mOCTN2 mRNA and protein levels, and improved renal impairment in this model. CONCLUSION Wudang cherry processes uricosuric and nephroprotective actions by regulating renal organic ion transporters in hyperuricemic mice.
9.Study on genotypes of plasmid-mediated quinolone in Shigella
Tieli ZHOU ; Mingming ZHOU ; Lizhen SHEN ; Fei XU ; Chao LI ; Jingxian FEI
Chinese Journal of Microbiology and Immunology 2009;29(8):691-694
ored carefully.
10.Optimization of reference genes for microRNA expression analysis in a mouse model of oxygen-induced retinopathy
Qiyu, BO ; Yu, ZHOU ; Xiaorong, LI ; Yan, ZHANG ; Fei, WANG
Chinese Journal of Experimental Ophthalmology 2015;33(2):102-108
Background Retinal neovascularization varies with the change of microRNA (miRNA) expression level.Quantitative real-time PCR (qRT-PCR) is a common method for the analysis of miRNA expression profiling.However,the housekeeping genes selected as references may exhibit differential expression levels under the distinctive experimental conditions.The accuracy of the levels of target gene expression often is affected if the selected housekeeping genes with significantly fluctuating expression as references.Determining a reference gene with stable expression level is the premise to consecutive studies.Objective This study was to compare the expression levels and stability of the frequently used reference genes for miRNA expression analysis in normally developing eyes and in eyes of a mouse model of oxygen-induced retinopathy (OIR),and select the optimal reference gene (s) exhibiting stable ocular expression under both hypoxia and normal development conditions.Methods P7,P12,P17,P37 and 8-week-old clean C57BL/6J mice were selected randomly.q-PCR was used to detect the dynamic changes of relative expressions of 5 kinds of reference genes in different ages of mice,including snRNAU6 (RNU6),5S ribosomal RNA (5s rRNA),snoRNA U68 (U68),snoRNA U70 (U70),snoRNA U49A (U49A).Other 40 P7 mice were randomized into the normal control group and the OIR group.The mice of the normal control group were fed in the normal oxygen environment,and those in the OIR group were raised in the (75-±2)% oxygen environment for 10 days.Fluorescine was injected via ritrobulbar vein for retinal stretched preparation,and the histopathological examination of mouse retinas was performed to identified the models.The expressing difference of 5 kinds of reference genes in the retinas were detected to compare the differences of 5 kinds of genes expression between the two groups.GeNorm program was employed to compare the stability of the expressing genes.This study were approved and granted by Ethical Committee of Tianjin Medical University and met the requirements stipulated in the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.Results Across the developmental ages,the expression levels of U68 in the eye exhibited the greatest variability,and then coming with U49A,U70,RNU6 and 5s rRNA.Significant differences were seen across the developmental ages for the expression levels of U68,U49A,U70,and RNU6 (U70:F =7.005,P =0.000 ; U68:F =10.189,P =0.000 ; U49A:F =21.134,P=0.000;RNU6:F=4.968,P=0.004).Expression of 5s rRNA showed the least variability across the developmental ages (F=2.099,P =0.107).In P17 mice of the OIR group,tortuous access vessels and non-perfusion area were found in the retinal section.Hematoxylin-eosin stain showed the neovascular sprout through across the inner limiting membrane.The relative expression of U70 exhibited the greatest variability and then were U49A,U68 and 5s rRNA in turn in P17 OIR mice,and significantly elevated expressions were found in U70,U49A,U68 and 5s rRNA genes between the OIR group and the normal control group (t =5.174,P =0.000;t =3.376,P =0.012;t =4.802,P =0.000 ;t=2.856,P=0.029).Expression of RNU6 showed the least variability between the two groups (t =2.104,P=0.065).As analyzed by GeNorm program,the stability for the five reference genes across developmental ages was 5s rRNA/RNU6> U70 > U49A > U68,and the optimal reference combination was 5s rRNA and RNU6.Whereas the stability for the OIR model was 5s rRNA/RNU6>U68> U49A>U70,and the optimal reference combination was 5s rRNA and RNU6.Conclusions Reference genes should be selected based on specific subjects and experimental conditions when qRT-PCR is used to analyze miRNA expression levels.In the OIR model,both developmental and hypoxic factors need to be considered.5s rRNA and RNU6 reference combination is the preferred option for the qRT-PCR analysis of ocular miRNA expression if available.