1.RNA Interference and Its Application to Metabolic Engineering in Plants
Hui LI ; Tian-Long WU ;
China Biotechnology 2006;0(10):-
RNA interference(RNAi) is an high efficient,specific gene silencing.The discovery process of RNAi and its mechanism was reviewed.In the further,RNAi will play important roles in characterizing new genes in the metabolic pathways,improving plant nutritional value,enhancing plant resistance and creating new varieties.
2.Correction of penile curvature by allogeneic acellular dermal matrix
Yulin DONG ; Xiaowei WU ; Long TIAN
Chinese Journal of Urology 2012;33(2):146-148
ObjectiveTo evaluate the effect of lengthening the short-side albuginea of the bending penis by the allogeneic acellular dermal matrix ( Allo-ADM ) for the treatment of penile curvature.MethodsFrom Jun 2007 to Jun 2010,18 patients with penile curvature due to malformation of the albuginea cavernous body were treated.The age of the patients ranged from 15 to 26 years (mean,20 years).Twelve patients were married.The curvature degree ranged from 30° to 80° (mean,55°).There were 17cases of single curvature and 1 case of complex curvature.The grafts ( Allo-ADM ) of different sizes were sutured to the albuginea at the curvatus side of the penis according to the extent of penile curvature through a circumcision incision.The extent of penile curvature and complications were evaluated postoperatively.ResultsPenile curvature was corrected in all 18 patients after the operation.No infection,hematoma and abnormal erection occurred postoperatively. No erectile dysfunction and penile re-curvature was observed during the follow-up period of 3 to 24 months.ConclusionLengthening the short-side albuginea of the bending penis by Allo-ADM could be a safe and effective way to correct penile curvature.
3.Interferon-induced Protein 204 Over-expression Inhibits Aortic Vascular Adventitial Fibroblast Proliferation and Migration in Experimental Rats
Fang SONG ; Maobo TIAN ; Yan XIAO ; Xiangshu LONG ; Qiang WU
Chinese Circulation Journal 2015;(11):1110-1114
Objective: To observe the effects of interferon-induced protein 204 (p204) over-expression on apoptosis, proliferation and migration of aortic vascular adventitial ifbroblast (VAFs) in experimental rats.
Methods: Our research included in 3 groups: Iif204-Lv group, in whichVAFs were infected by Iif204-recombined lentivirus, Con-Lv group, in which VAFs carried the empty vector without virus, Blank control group, in which VAFs were untreated. VAFs proliferation was examined by MTT method, cell apoptosis was measured by lfow cytometry and the migration was detected by scratching assay and transwell chamber method. The mRNA and protein expressions of p204, p53 and p21were evaluated by real-time q RT-PCR and Western blot analysis respectively.
Results: Compared with Con-Lv and Blank control groups, Iif204-Lv group had decreased VAFs proliferation (by OD value) at 48 hours: (0.53 ± 0.05) vs (0.66±0.03) and (0.63 ± 0.06), at 72 hours: (0.89 ± 0.06) vs (1.02 ± 0.06) and (1.01 ± 0.07); distance of cell migration (by pixel): (61.00 ± 1.83) vs (74.50 ± 6.25) and (75.50 ± 7.85); number of cell migration: (61.75 ± 10.69) vs (155.25 ± 10.21) and (153.75 ± 9.40), allP<0.05. VAFs apoptosis rates were similar among different groups. Compared with Con-Lv and Blank control groups, Ifi204-Lv group presented up-regulated mRNA expressions of p204 (3.45 ± 0.15) vs (2.09 ± 0.10) and (2.06 ± 0.09); p53 (3.41 ± 0.09) vs (2.06 ± 0.07) and (2.10 ± 0.06); p21 (3.01 ± 0.08) vs (2.05 ± 0.06) and (2.11 ± 0.08), allP<0.05.
Conclusion: p204 over-expression inhibits VAFs proliferation and migration which might be partly related to the activation of p53 and p21 expression in experimental rats.
4.Interferon alpha suppresses proliferation and induces apoptosis of human brain vascular adventitial fibroblasts via IFI16
Jing HUANG ; Fang SONG ; Xiangshu LONG ; Maobo TIAN ; Qiang WU
Chinese Journal of Immunology 2017;33(4):494-497
Objective:To study interferon alpha (IFN-α) inhibition of proliferation and apoptosis induction of human brain vascular adventitial fibroblasts(HBVAFs)via IFI16.Methods:Cultured HBVAFs were treated with transfection IFI16 siRNA and/or IFN-α in vitro instantaneously.The protein and mRNA levels of IFI16,P53,P21 were measured by Western blot and Real-time PCR.MTT was used to detect the cell proliferation of the HBVAFs.Cell cycle and apoptosis were analyzed by flow cytometry.Results:IFN-α with terminal concentration of 2 000-5 000 kU/L induce significantly expression of IFI16 in HBVAFs,without any significant difference.Stimulated with 2 000 kU/L IFN-α up-regulated the expression of P53,P21 at protein and mRNA levels,and inhibited the cell proliferation and promote cells apoptosis in HBVAFs.Such effect was restrained by transfection with IFI16 siRNA into HBVAFs.Conclusion:IFN-α inhibits HBVAFs proliferation and induces apoptosis may partly relate to the increased IFI16 expression.
5.Research progress of new isothermal amplification technology in rapid diagnosis of Plasmodium and other parasites
YANG Jun-jie ; WU Tian ; LIU Cheng-long ; LIAN Meng-ze ; ZHANG Yi-long
China Tropical Medicine 2022;22(09):884-
Abstract: At present, nucleic acid detection technology based on the PCR principle is commonly used to detect malaria parasites, the existing Plasmodium detection methods mainly include microscopy, antigen immunoassay, and nucleic acid detection,but due to the long detection time, high personnel and equipment requirements, and other shortcomings, its popularization, and application at the grassroots level are limited. What challenges previous Plasmodium detection methods are the lack of experienced professionals and advanced equipment at the grassroots as well as the requirement of rapid detection of large samples under extreme conditions. The isothermal amplification technology developed in recent years has potential application prospects due to its simplicity, rapidity, high sensitivity, and high specificity. This article attempts to review the principles, characteristics, and prospects of various isothermal amplification technologies, and on this basis, focuses on the introduction of recombinase polymerase amplification (RPA) and recombinase⁃aided isothermal amplification (RAA) assay technologies and proposes the use of such recombinant enzyme amplification technologies to achieve rapid and accurate diagnosis of common Plasmodium species possibility and imagination.
6.Regulation of deleted in liver cancer-1 gene domains on the proliferation of human colon cancer HT29 cell
Pingping WU ; Peng WU ; Qiqiang LONG ; Nan LI ; Zhi JIN ; Xiaoqiang TIAN ; Peilin HUANG
Chinese Journal of Digestion 2012;32(11):744-749
Objective To study the role of deleted in liver cancer-1 (DLC-1) gene main domains on the regulation of human colon cancer HT29 cell proliferation.Methods Subcloning recombinant plasmid vectors with Rho GTPase activating protein (RhoGAP),sterile alpha motif (SAM) or steroidogenic acute regulatory-related lipid-transfer (START) domains of DLC-1 gene knockout were constructed and transfected into human colon cancer cell HT29.Wild HT29 cell group (control group),pcDNA3.1-HT29 cell group (vector group) and pcDNA3.1-HT29-DLC-1 cell group (whole DLC-1 gene transfected group) were set as control.The change of cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and colony formation test.The cell apoptosis was analyzed by flow cytometry.The activity of RhoA protein was detected by pull-down assay.The differences between the groups were analyzed by the analysis of variance.Results At 48 hours after the successful transfection,compared with control group and vector group,cells proliferation and the activity of RhoA protein were significantly suppressed in whole DLC-1 gene transfected group (F=146.36,698.08,both P<0.05) and early cell apoptosis increased (F=294.08,P<0.05).Compared with control group and vector group,there was no significant difference in cell proliferation ability,cell apoptosis and the activity of RhoA protein activity in RhoGAP knockout transfected cells (F=0.99,0.049,5.769,all P>0.05).Compared with whole DLC-1 gene transfected group,the suppression of cell proliferation was more significant in SAM knockout transfected cells (F=31.00,P<0.05),the activity of RhoA protein down regulated (F=92.57,P<0.05) and apoptosis increased (F=130.44,P<0.05).Compared with whole DLC-1 gene transfected group,the ability of cell proliferation increased (F=15.47,P<0.05),apoptosis cell decreased (F=110.23,P<0.05) and the activity of RhoA protein up regulated (F=199.39,P<0.05) in START knockout transfected cells.Conclusions The role of DLC-1 gene in the suppression of cell proliferation in HT29 cells was RhoGAP-dependent.SAM domain may be the self suppression domain for endogenous RhoGAP activity.START domain may take effect through enhancing RhoGAP domain.
7.Differentiation of Spinal Neural Stem Cells from Various Gestational Aged Fetal Rats
Wei NAN ; Dongliang FENG ; Kaisheng ZHOU ; Yandong ZHU ; Jing WANG ; Ting TIAN ; Long JIANG ; Zaiyun LONG ; Yamin WU ; Haihong ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(10):1145-1150
Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.
8.Study on the correlation between contrast-enhanced ultrasound perfusion imaging typing and vascularization
Qi WANG ; Gang WU ; Long YANG ; Jianjun YUAN ; Yuanrui HAO ; Jianguo TIAN
Chinese Journal of Ultrasonography 2010;19(1):25-28
Objective To study the significance and value of imaging typing by analyzing the correlation between contrast-enhanced ultrasound perfusion imaging typing and vascularization in patients with primary hepatocellular cancer.Methods The early enhanced arterial phase reflecting angioarchitecture,and the size and edge of enhanced tumor closely related to microvessel distribution, position and density, were observed and analyzed in 89 patients (113 lesions).All the cases were pathologically proved, and some received immunohistochemisty staining and the microvessel density were recorded.Results The perfusion imagings were classified into 4 types according to changes on contrast-enhanced ultrasound,dendritic,mixed,circular and net-like type.Atypia vessels were commonly seen in dendritic type,and Ⅲ and Ⅳ accounted for 79.1%.The microvessel density was higher than the other types.The circular type was relatively regular and Ⅲ and Ⅳ accounted for 12.5%.The microvessel density was lower than the other types.Combined with the contrast-enhanced ultrasound perfusion imaging and compared with pathological grading,growing methods and microvessel density, the dendritic type was characterized by infiltrative growth, and had strong invasion tendency.The circular type was characterized by expansive growth.And the other two types were characterized by transitional type.Conclusions Contrast-enhanced ultrasound can reflect the angioarchiteeture, the microvessel density and the position,and it is related to the pathological grading and growing methods.
9.Inducement Effect of Interferon Alpha on Apoptosis of Rat Vascular Smooth Muscle Cells via P204 and RAS Signal Pathway
Xiangshu LONG ; Qiang WU ; Taihe LIU ; Fang SONG ; Jing HUANG ; Maobo TIAN ; Yan XIAO
Herald of Medicine 2015;34(12):1555-1558
Objective To investigate the effect of interferon alpha ( IFN-α) on apoptosis of vascular smooth muscle cells ( VSMCs) in rats and the related mechanism. Methods The cells were divided into three group:group A, group B and group C.Group A was transfected with nonspecific siRNA, group B was intervened with IFN-α and transfected with nonspecific siRNA, and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).P204, RAS protein levels, and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC/PI method. Results As compared with group A, the expression of P204 mRNA and protein in group B was up-regulated (P<0.05), the cell apoptosis was increased (P<0.05), in the process of the above, the expression of RAS protein was decreased ( P<0.05) and the phosphorylation levels of RAF and ERK were dropped (P<0.05).In group C, the expression levels of P204 mRNA and protein were down-regulated (P<0.05), and cell apoptosis was decreased ( P<0.05) , the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased ( P<0.05) . Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.
10.Relationship Between Body Weight Changes From Early Adulthood to Middle Age and the Prevalence of Hypertriglyceridemia by Large-scale Cardiovascular Risk Factor Investigation
Long ZHOU ; Ying LI ; Min GUO ; Ye TIAN ; Yangfeng WU ; Liancheng ZHAO
Chinese Circulation Journal 2015;(10):962-966
Objective: To explore the relationship between body weight changes from early adulthood to middle age and the prevalence of hypertriglyceridemia by large-scale cardiovascular risk factor investigation.
Methods: A total of 15 population groups from China multi-center collaborative study of cardiovascular epidemiology in 1998 were enrolled. There were approximately 1000 participants in each group including 50% of each male and female at the age of (35-59) years which was deifned as middle age. The participants were surveyed for cardiovascular risk factors including the body weight and body mass index (BMI) at the age of 25 years which was deifned as early adulthood. The participants were divided into 4 groups based on BMI at early adulthood: Low body weight group, BMI < 18.5 kg/m2, Normal body weight group, BMI (18.5-23.9) kg/m2, Overweight group, BMI (24-27.9) kg/m2 and Obese group, BMI ≥ 28 kg/m2. The changes of body weight from early adulthood to middle age were calculated by body weight at present minus body weight at 25 years; according to the differences, participants were divided into another set of 6 groups: differences < -7.5 kg, (-7.5 to -2.6) kg, (-2.5 to 2.5) kg, (2.6 to7.5) kg, (7.6 to 12.5) kg and > 12.5 kg. The relationship between body weight status at 25 years of age with subsequent changes and the prevalence of hypertriglyceridemia were investigated.
Results: A total of 13883 participants finished the investigation.①The prevalence of hypertriglyceridemia by 4 BMI groups were at 22.8%, 26.0%, 27.4% and 30.8% respectively (the trend ofP<0.01).②The prevalence of hypertriglyceridemia by 6 age differences groups were at 12.5 %, 14.0 %, 17.8 %, 24.2 %, 31.5 % and 40.9 % respectively (the trend ofP<0.01). Multivariate Logistic regression analysis indicated that overweight and obesity at the age of 25 years with subsequent weight gain were positively related to the risk of hypertriglyceridemia at the middle age (the trend ofP<0.01).
Conclusion: Overweight and obesity in early adulthood with subsequent weight gain were independently related to the risk of hypertriglyceridemia at the middle age in our survey.