2.Functions of long noncoding RNAs and their roles in ocular diseases
Fang-Kun, ZHAO ; Yu, QIN ; Jing, LI ; Jin-Song, ZHANG
International Eye Science 2016;16(8):1469-1473
?Advances in genome-wide analysis have revealed that up to 90%of the human genome is transcribed.However, only approximately 1% of RNA transcripts encode proteins, and the remaining transcripts are noncoding RNAs.Noncoding RNAs can be roughly divided into small noncoding RNAs (<200nt ) and long noncoding RNAs ( LncRNAs, >200nt ). Small noncoding RNAs include microRNAs, transfer RNAs and small nucleolar RNAs, whereas the long noncoding RNAs comprise ribosomal RNA, natural antisense transcripts, etc. Although the biosynthesis and biological activities of microRNAs are well studied through bioinformatics and active biological molecules analysis, the understanding of LncRNAs on these aspects is still limited.LncRNAs play multiple roles in regulating gene transcription and translation, and epigenetics.Aberrant LncRNAs expression can occur in various pathological processes and significantly related to the pathogenesis or poor prognosis of ophthalmological diseases. In this review, we will focus on the characteristics and regulatory functions of LncRNAs that are commonly associated with ophthalmological diseases.
3.Expression of Plasminogen Activator Inhibitor-1 of Frozen Muscle Specimensin Muscular Dystrophy
gui-lian, SUN ; hong-kun, JIANG ; shuang, ZHAO ; jing, ZHANG
Journal of Applied Clinical Pediatrics 2006;0(24):-
Objective To explore the role of plasminogen activator inhibitor-1(PAI-1)in development of progressive fibrosis via the inhibition of extracellular matrix degradation,and to reveal the contributive role of PAI-1 in muscular dystrophy(MD).Methods Expression and cellular localization of PAI-1 protein were examined in frozen muscle specimens obtained via biopsy from 5 patients with duchenne muscular dystrophy(DMD),3 patients with becker muscular dystrophy(BMD),9 patients with congenital muscular dystrophy(CMD) and 4 cases with normal muscle by immunohistochemistry,double immunofluorescence and Western-blot analysis.Results PAI-1 was positive only in vascular endothelial cells of normal muscle.Both immunohistochemistry and Western-blot analysis showed that PAI-1 expression distinctly increased in most dystrophic muscles of MD than that in normal muscles.Double immunolabeling revealed that PAI-1 strongly expressed in cytoplasm and nuclei of regenerating muscle fibers,macrophages,macrophage infiltrating necrotic fibers.Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were positive for PAI-1.Conclusions The functional consequence of overexpression of PAI-1 in dystrophic muscles is unknown but the elevated local expression of PAI-1 in diseased muscles of MD and their distinct distribution pattern provide evidence that PAI-1 participate in pathogenesis of MD.
4.A Meta-analysis of the related factors of PICC malposition
Lin LI ; Xiaodan LIU ; Jing REN ; Kun LIU ; Qianqian ZHANG
Chinese Journal of Practical Nursing 2014;30(30):14-19
Objective To identify the related factors of PICC malposition.Methods The domestic published literature about the related factors of PICC malposition were collected from 2001 to 2014.Analysis of heterogeneity was performed and cumulative effects were calculated using either fixed or random effects models by RevMan 5.0.Results The incidence of PICC malposition in patients with median cubital vein or cephalic vein was obviously higher than that in patients with basilic vein,the incidence of PICC malposition in patients with cephalic vein was significantly higher than that in patients with median cubital vein; the incidence of PICC malposition in patients with recumbent position was significantly higher than that in patients with seat position; the incidence of PICC malposition in the older than sixty years patients was significantly higher than that in younger than sixty years patients; the incidence of PICC malposition in male and female patients was not discrepant.Conclusions The vein,position and age of patient are the related factors of PICC malposition,while sex factor has nothing to do with the occurrence of PICC malposition.
5.Vaspin promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in rats through the Wnt/β-catenin signaling pathway
Fei GAO ; Kun CHEN ; Yan ZHANG ; Jing YANG
Chinese Journal of Endocrinology and Metabolism 2016;32(6):505-509
_ Objective_ To observe the effect of vaspin on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells in rats and its potential mechanism. Methods Bone marrow mesenchymal stem cells( BMSCs) from 4 weeks aged rats were isolated and cultured. The BMSCs were treated with osteogenic induction medium and different concentrations of vaspin and Wnt signaling pathway inhibitor DKK1. The proliferation was detected by CCK8 method at 24, 48, 72 h. After 7 days, the mRNA expressions of ALP, Runx2,β-catenin were detected by realtime qPCR. The expression levels of Runx2 and β-catenin protein were detected by Western blot. After 21 days, alizarin red stained mineralized nodules and quantitative detection were performed. Results Vaspin had no effect on the proliferation but promoted the expression of osteogenic differentiation gene ALP, Runx2, and also increasedβ-catenin mRNA and expression of Runx2 and β-catenin protein. Mineralized nodules were brown and increased, OD value of vaspin group was higher than control group. After adding the DKK1, the expression of ALP, Runx2,β-catenin mRNA and Runx2, β-catenin protein were significantly decreased(all P<0. 05). Conclusion Vaspin can promote BMSCs into osteogenic differention through the Wnt/β-catenin signaling pathway.
6.A comparison study of axillary small incision and assisted thoracic surgery for spontaneous pneumothorax
Kun GAO ; Ge ZHANG ; Huagang LIANG ; Xiaochun JING
Chinese Journal of Primary Medicine and Pharmacy 2013;20(6):828-829
Objective To study the clinical efficacy of axillary incision and thoracoscopic surgery for spontaneous pneumothorax.Methods 106 cases of spontaneous pneumothorax in our hospital were given axillary incision surgery(axillary incision group) and thoracoscopic surgery(thoracoscopy group).The intraoperative blood loss,operative time,chest tube drainage time,postoperative hospital stay and surgery costs were compared between the two groups,and the occurrence of complications were observed.Results The armpit small incision group,intraoperative blood loss was (44.5 ± 5.2) ml,the thoracoscopic amount of blood loss was (38.3 ± 6.5) ml (t =6.378,P < 0.01) ;armpit operation time of the small incision group was (68.0 ± 5.3) min,thoracoscopic operative time was (60.8 ±6.0)min; armpit chest tube drainage time of small incision group was (2.8 ± 0.8)d,thoracoscopic group of chest tube drainage time was (2.0 ± 0.5) d; axillary small incision group,length of stay was (4.8 ± 0.7) d,the thoracoscopic group hospitalization time was (4.0 ± 0.6) d,(t =3.552,4.215,3.076,all P < 0.05) ; axillary incision surgery costs was (1 550 ± 348) Yuan,the thoracoscopic group cost of surgery was (4 290 ± 573) Yuan (t =-24.823,P < 0.05).Two groups of patients with no surgical complications,chest X-ray review of lung reexpansion good thoracoscopic group one cases of recurrence of pneumothorax,axillary incision group without recurrence (P > 0.05).Conclusion Axillary small incision and thoracoscopic surgery for spontaneous pneumothorax have the similar efficacy,thoracoscopic surgery is less trauma,faster recovery,shorter hospital stay,but the high cost of surgery,if patients physical condition is acceptable,which can be used axillary incision surgery.
7.Effects of silencing survivin gene by RNA interference on cell apoptosis using ultrasound targeted microbubble destruction techniques
Zhiyi CHEN ; Kun LIANG ; Mingxing XIE ; Jing ZHANG
Tumor 2009;(7):626-630
Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.
9.Research on Exogenous Gene mRNA Levels in Zymomonas mobilis by Semi-quantitative RT-PCR
Yuanyuan MA ; Shaolan ZOU ; Kun ZHANG ; Jiefang HONG ; Xin JING ; Cheng LIU ; Minhua ZHANG
Microbiology 1992;0(06):-
An effective RT-PCR method was developed to detect exogenous gene xylB transcript levels in Zymomonas mobilis CP4. Total RNAs without genomic DNA contamination were purified from wild-type and gene engineering strains, and were quantified to the same concentration. Then, cDNAs synthesis and PCR analysis of these samples were conducted by reverse transcription PCR. The optimal number of cycles was determined by observing amplification profile of target gene xylB and internal control gene 16s rRNA, and relative expression levels of xylB in various samples were analyzed by RT-PCR. The results indicated that the xylB transcript was not be detected in CP4, however that could be found in recombinant strains, in which xylB transcription abundance was similar. The enzyme assay furthermore confirmed that effective ex-pression of the target gene. The method provided a useful and rapid tool for detecting transcript levels of target genes from various samples of Z. mobilis.
10.Study on the relationship between rapid aging and avoidance of tumor in senescence-accelerated mouse
Kun NIE ; Yujie JIA ; Jing LI ; Xuezhu ZHANG ; Zhen ZHANG ; Yao WANG
Chinese Journal of Geriatrics 2017;36(7):793-797
Objective To investigate the role of p53 in the regulation of heat shock protein(Hsp)84 and 86,and the correlation of their functional imbalances with accelerated brain aging and with suppressed tumorigenesis in SAMP8 mice(senescence accelerated mouse prone 8).Methods The mRNA and protein expressions of Hsp84 and Hsp86,and protein expressions of p53 pathway-related proteins(p21 and MDM2)in hippocampus of SAMP8 mice and their control SAMR1(senescence accelerated mouse resistant 1)mice were determined.Murine Neuro-2a cells were treated with 20 μmol/L Aβ25-35,and then mRNA expressions of p53,Hsp84 and Hsp86 in these cells were detected.Neuro-2a cells were co-transfected with p53 siRNA and pHsp84-Luc or pHsp86-Luc plasmid and treated with 20 μmol/L Aβ25-35,then promoter activity of Hsp84 and Hsp86 were detected in these cells.After co-transfection with pcDNA3.1-p53 or pcDNA3.1-p53DD and pHsp84-Luc or pHsp86-Luc plasmids,the neuro-2a cells were treated with 20 μmol/L Aβ25-35.Then promoter activity of Hsp84 and Hsp86 were detected in these cells at different concentrations of p53.Results The mRNA levels of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were significantly declined,which were 13.51% and 16.13% of SAMR1 mice,respectively(all P<0.01).Compared with the SAMR1 mice,the protein expressions of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were obviously declined(all P<0.01).Whereas,p53 pathway-related protein p21 expression was increased and MDM2 expression was decreased(all P<0.01).The mRNA expression of p53 in AD cells was significantly increased by 58%(P<0.01),whereas Hsp84 and Hsp86 mRNA levels were significantly decreased by 32% and 41%,respectively as compared with the normal cells(all P<0.05).Inhibition of p53 in AD cells could increase promoter activity of Hsp84 and Hsp86 significantly in a concentration-dependent manner(both P<0.05),whereas overexpression of p53 in the cells could lead to decreased promoter activity of them in a concentration-dependent manner(both P<0.05).Conclusions The p53 can negatively regulate the expressions of Hsp84 and Hsp86.The activity of p53/p21 pathway is increased,while Hsp84 and Hsp86 are inhibited in the brain of SAMP8 mice.Functional imbalance between p53 and Hsp84/86 might be the part of reasons causing accelerated aging and suppressed tumorigenesis in SAMP8 mice.