1.Construction of a stable mouse fibroblast line targeting mammalian target of rapamycin by lentivirus mediated short hairpin RNA
Xiao, SUN ; Xuejuan, CHEN ; Chen, ZHAO ; Kanxing, ZHAO
Chinese Journal of Experimental Ophthalmology 2014;32(8):688-692
Background Age-related macular degeneration (AMD) is a group of vision-threatening eye disorders.Previous researches showed that the activation of Akt/mammalian target of rapamycin (Akt/mTOR) pathway closely related to the mechanism of AMD.A new specific method to inhibit Akt/mTOR pathway will become a breakthrough for the treatment of AMD.Objective This study was to establish a mouse fibroblast cell line (NIH/3T3) which can stably inhibit the expression of mTOR gene and provide a cell model for the study on the function of Akt/mTOR pathway in AMD and observe the influence of mTOR gene knockdown on related proteins.Methods Three short hairpin RNAs (shRNAs) targeting mTOR gene were designed and synthesized based on murine mTOR mRNA sequence.Double-strand shRNA hairpins were separately cloned into PIGZ-green fluorescent protein (GFP) +Puro vectors to produce recombination plasmids.The packaged lentiviral plasmids and RNAi plasmids were co-transfected into NIH/3T3 cells,a mouse fibroblast line.After puromycin selection and culture expansion,0.5 mg/L puromycin was added the culture medium to establish stable cell clones.The expressions of mTOR mRNA and protein in NIH/3T3 cells were detected by real-time PCR and Western blot respectively,and the inhibitory efficiency of interference was analyzed.Results Transfected GFP-labeled NIH/3T3 cells by lentiviral presented the green fluoresccence with the efficiency of infection of 90% in the third day.Real-time PCR showed a distinct band of mTOR mRNA in 184 bp.The knockdown rate for sh1,sh2 and sh3 were respectively 31.3%,31.8% and 45.3% in the lower multipolicity of infection (MOI) group ;while in the higher MOI group,the knockdown rate for sh1,sh2 and sh3 were 47.1%,56.5% and 71.6% respectively.Western blot assay exhibited weakened expression band of mTOR protein in NIH/3T3 cell line for sh1,sh2 and sh3 in both lower and higher MOI groups with the weakest expression for sh3.Conclusions A stable mouse fibroblast cell line is established by the inhibition of mTOR with RNA interference technique,which can provide a cell model for studying the function of Akt/mTOR pathway in AMD.
2.Myokines and osteokines in muscle-bone interactions
Dongmei CHEN ; Kexiang ZHAO ; Qian XIAO
Chinese Journal of Geriatrics 2017;36(3):344-347
Sarcopenia and osteoporosis are two common diseases with adverse effects on the health of the elderly and share many characteristics.As target organs of the two diseases,muscles and bones are closely connected,not only in anatomy and physiology,but also in pathophysiological processes.Initially,mechanical stimulation was conducted to explain the relationship between muscle and bone at the macroscopic level.Currently,research is increasingly focused at the cellular and molecular levels.Both muscle and bone tissues can secrete a number of growth factors,cytokines,polypeptides,and so on.Myokines include fibroblast growth factor-2 (FGF-2),myostatin,irisin,insulinlike growth factor-1 (IGF-1),interleukins,musclin,among others.Osteokines comprise osteocalcin,fibroblast growth factor-23,IGF-1,vascular endothelial growth factor,etc.In this review,we summarize the roles of myokines and osteokines in muscle-bone interactions.
3.Determination of Anionic Surfactants in Waters by On-line Double Microporous Membrane Extraction-Flow Injection Spectrophotometry
Ping ZHAO ; Jinhui CHEN ; Jingze XIAO
Chinese Journal of Analytical Chemistry 2009;37(12):1853-1856
Anionic surfactants in waters were determined using on-line double microporous membrane extraction-flow injection analysis method. This method was based on the extraction of the ion-association complex (λ_(max)=650 nm) which formed from methylene blue cation and anionic surfactants. The on-line double microporous membrane liquid-liquid extraction phase separators were adopted. Optimization of variables of the flow injection manifold, groove mechanical dimension of the phase separator, and pore size of the membrane was performed. The proposed method had a linear range of 25.0-1000.0 μg/L(r≥0.999). The detection limit was 4.28 μg/L. The relative standard deviation(n=7) of different concentration was 0.7%-6.0%. The recovery was between 96%-110% and the sample throughput was 18 h-1. The method was applied to the analysis of standard Reference materials with satisfactory results. For practical application, two water samples were analyzed with the established method.
4.Congenital hepatic fibrosis: report of a case.
Chinese Journal of Pathology 2010;39(7):485-486
5.Characteristics of human bocavirus infection in children in Chongqing area.
Yuan DING ; Xiao-dong ZHAO ; Xin CHEN
Chinese Journal of Pediatrics 2009;47(6):466-467
Child
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China
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epidemiology
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Female
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Human bocavirus
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Humans
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Infant
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Infant, Newborn
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Male
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Parvoviridae Infections
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epidemiology
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virology
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Respiratory Tract Infections
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epidemiology
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virology
6.Psoriasis patients in china: socio-demographic and clinical characteristics at different disease onset age
Yi XIAO ; Shuang ZHAO ; Yue ZHAO ; Wu ZHU ; Xiang CHEN
Journal of Chinese Physician 2016;18(4):544-547,551
Objective To explore different socio-demographic and clinical characteristics for early onset patients (EOP) with psoriasis and late onset patients (LOP) with psoriasis in Chinese population and to provide scientific evidences for establishing comprehensive prevention and treatment strategy for psoriasis.Methods Cross-sectional study was performed.A total of 942 diagnosed psoriasis patients who paid a visit to outpatient clinic in a tertiary hospital in China from 12th September 2013 to 19th May 2015 was consecutively enrolled and investigated.Exploratory analysis was conducted to detect the association between disease onset age and patients features.Results Among 942 enrolled psoriasis outpatients,the average age was (40.6 ± 13.6) years with a range from 5 to 80 years.The sex ratio was 2.1 in favor of male.The most observed type of psoriasis in the present study was psoriasis vulgaris (98.3%).The average psoriasis area and severity index (PASI) for patients were 9.47 ±7.63.Comorbidity was combined in 20.2% cases.Patients with EOP had a significantly higher likelihood of family history of disease.Compared EOP to LOP,LOP had a significantly higher likelihood of comorbidities involvement (P < 0.05) and significant higher BMI index (P < 0.05).Conclusions The present study supports the hypothesis that there are clinical differences between EOP and LOP in Chinese population.Both dermatologists and patients should pay more attention to psoriasis-associated features,such as family history and comorbidity diseases involvements.
7.Control study of the ultrasonography with the placental vascular casts of the acardiac twins sequence
Xinlin CHEN ; Xiaohong YANG ; Peiwen CHEN ; Sheng ZHAO ; Qiaoying PENG ; Mei XIAO ; Xiaojun LU ; Lei ZHAO
Chinese Journal of Ultrasonography 2011;20(4):327-330
Objective To investigate the relationship between the ultrasonography diagnosis in prenatal and the postnatal placental vascular casts of the twins reversed arterial perfusion(TRAP) sequence. Methods Six cases of TRAP sequence underwent the two-dimensional and color Doppler ultrasonography, and performed placental vascular casts after termination of pregnance. Results According to the anatomical features and ultrasonography, there were 3 cases of acardius acephalus( IV) ,2 cases of acardius anceps( II ) , and 1 case of acardius acormus( I ). In all 6 cases,there were arterio-arterial and veno-venous anastomoses at the level of the chorionic plate between the normal co-twin and the acardiac twin,these anastmosis vessels were superficial and large calibre, and the distances of the placental insertions of the umbilical cords were different,4 cases <0. 5 cm, 2 cases were 4-5 cm. In 4 cases, the vessels entered and went out of the acardiac twin were just one large umbilical artery and one large umbilical vein. In 2 cases, the umbilical artery and umbilical vein had many vascular branches in the placenta. Out of 6 cases of the pump twin, 4 cases died,and 2 cases survived. Conclusions Ultrasonography was consistent with the placental vascular casts,the contrast observing gives us new information for the further research of the special type of the vascular anastmosis and prognosis of the TRAP sequence.
8.Transurethral resection of the hyperplaitic prostate using bipolar plasmakinetic technique
Tao ZHAO ; Weizhong XIAO ; Jianhua ZHANG ; Yong CHEN
Journal of Third Military Medical University 2003;0(14):-
Objective To assess the efficacy and safety of transurethral bipolar plasmakinetic resection of benign prostatic hyperplasia (BPH). Methods A total of 45 cases of BPH were treated by transurethral bipolar plasmakinetic resection of prostate (PKRP) and were followed up for 3-10 months.Results The weight of the resected prostate was 28-105 g with an average of (47?16) g, the operation time was 15-120 min with an average of (50?12) min. No case needed blood transfusion during the operation and no transurethral resection syndrome occurred. The mean catheterization time was 4 d and the mean postoperative hospital stay was 7 d. Peak urine flow increased from (7.3?1.5) ml/s to (16.2?4. 4) ml/s and IPSS decreased from (27.6?1.3) to (5.8?1.0) in 3 months postoperatively. Conclusion Transurethral bipolar plasmakinetic resection of prostate is effective and safe with less complications.
9.Assessment on changes of anterior chamber parameters after ICL surgery with Allegro Oculyzer system and their correlation with IOP
Qing, CHEN ; Peng, YU ; Ya-Li, ZHAO ; Xiao-Mei, GUO
International Eye Science 2016;16(8):1515-1518
Abstract? AIM: To investigate changes of anterior chamber parameters after posterior chamber phakic implantable collamer lens ( ICL ) surgery and its correlation with intraocular pressure ( IOP) .?METHODS: This was a retrospective case series study. Seventy four eyes in 43 myopia patients were examined by Allegro Oculyzer anterior segment tomography to obtain the changes of anterior chamber volume ( ACV ) , anterior chamber angle ( ACA) , central anterior chamber depth ( ACD) and vault, meanwhile, to measure the IOP to analyze the correlation with anterior chamber parameters.?RESULTS: Compared with preoperative, ACV, ACA, ACD all decreased apparently ( P <0.01 ) in the postoperative period of 1, 3, and 6mo.ACV, ACA and ACD showed their stability in each period after the surgery.There were changes of vault in 1 and 6mo ( t=27.66, P=0.01).IOP had not altered when compared with the preoperative.This research revealed that IOP had no relation with ACV, ACA, ACD and vault in every phase( all P>0.05).?CONCLUSION:For patients underwent ICL, the anterior chamber parameters all decreased which included ACV, ACA, ACD, and had stabilized since early postoperative period. Correspondingly, IOP was stable and had not correlate with ACV, ACA, ACD and vault, however the long-term observation is still necessary.
10.Investigation of Factors for Retinopathy of Premature Infants
li-jun, CHEN ; wen-jiang, CAI ; xiao-jian, ZHAO
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To investigate the factors for retinopathy in premature infants. Methods Ophthalmologic mydri was conducted in premature infants and acording to the international classification of ROP, they were diagnosed as ROP. The risk factors of the incidence of ROP were statistically and respectively analyzed. Results Among total of 47 patients, 11 patients were found retinopathy of prematurity. Conclusion The body weight is lower, the gestation is shorter, incidence of ROP is the higher.