1.The inhibition of hTERT and TRF2 gene expression and inducing cells apoptosis by adenovirus-mediated hTERT/TRF2 RNA interference in MCF-7 cells
Lan LIU ; Shaokun CHEN ; Qinglin SHUI ; Yongqiu ZENG ; Hong YU
China Oncology 2009;19(10):749-754
Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.
2.Exploration and practice of bilingual teaching in excellent doctor education and training
Lan LIU ; Yongqiu ZENG ; Jiao ZHAO ; Qinglin SHUI
Chinese Journal of Medical Education Research 2014;(9):902-905
Excellent doctor education training plan is an important policy applied by the gov-ernment to enhance the quality of medical personnel training in China. One of the important purposes is to improve the medical students' ability to apply and practice the medical professional English. In order to achieve this purpose, we have applied bilingual teaching for the students of excellent doctor in medical cell biology course. This article mainly introduces some reform measures , which involves training teacher, preparing textbooks and resources, establishing network database, exploring teaching modes, improving teaching methods and means, implementing diversified evaluation methods, and so on. At the same time, we have summarized some problems in the teaching process, and thus put for-ward improvement strategy.
3.Effects of L-Asparaginase Administration on Coagulation Function in Children with Acute Lymphoblastic Leukemia
wan-shui, WU ; gui-lan, LIU ; le-ping, ZHANG
Journal of Applied Clinical Pediatrics 2004;0(09):-
0.1).One(1.3%) of them suffered from intracranial hemorrhage.Conclusions 1.(L-ASP) may affect the function of coagulation system,such as prolonged APTT and hypofibrinogenemia.2.There was no statistical difference in side effects of coagulation,in use of domestic L-ASP and foreign L-ASP,intravenous dosing and intramuscular dosing.
4.Studies on macroscopic and microscopic characteristics of Ophiocordyceps xuefengensis.
Hao LIU ; Lan-ping ZHEN ; Ru-cai ZHU ; Shui-han ZHANG ; Hui-yong HUANG
China Journal of Chinese Materia Medica 2015;40(14):2820-2824
The macroscopic characteristics, tissue, caterpillar body wall and powder of Ophiocordyceps xuefengensis in different batch numbers were observed and researched by the macroscopic and microscopic identification methods. The result shows that the morphology, size, abdominal annulations of caterpillar, etc. of 0. xuefengensis are the macroscopic identification characteristics, the caterpillar body surface mycelium, body wall sculpture and crochets on abdominal legs are the microscopic identification characteristics. These characters are stable and regular discriminant features, which are proved to be the identification basis of O. xuefengensis. In addition, The characters such as crochets on abdominal legs arrange in two parallel ellipse rings, the inner crochets are long strip, and the external toes are unciform, are specific.
Animals
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Hypocreales
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cytology
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Moths
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anatomy & histology
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cytology
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microbiology
5.Effect of Intracerebral Transplantation of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood on Hypoxic-Ischemic Brain Damage in Neonatal Rat
gui-zhi, XIA ; xin-ru, HONG ; xin-min, CHEN ; shui-liang, WANG ; feng-hua, LAN
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To investigate the therapeutic effect of intracerebral transplantation of mesenchymal stem cells(MSCs) derived from human umbilical cord blood(UCB) on hypoxic-ischemic brain damage(HIBD) in neonatal rat.Methods Twenty samples of human UCB were collected from healthy full-term newborns.MSCs were isolated from human UCB by density gradient centrifugation and purified by adhere cell selection method.For transplantation,P3 human UCB-derived MSCs were labeled by the 5-bromo-2-deoxyuridine (BrdU).Thirty SD rats of 7 d were built for neonatal HIBD model.One rat died and others were divided into transplant group(n=18) and control group(n=11).At the third day after building models,human UCB-derived MSCs were injected into left cortex in transplant group,while PBS of the same volume was injected into the same site in control group at the same time.The seventh day after transplantation,6 rats of transplant group were sacrificed to prepare brain tissue sections.The survival,migration and differentiation of the transplanted cells were investigated by brain tissue immunohistochemical analysis,and nervous function of 2 groups were evaluated by modified neurological severity score(mNSS) on the first,7th,14th,21th and 28th day after transplantation.Results MSCs were isolated from 5 of 20 human UCB samples.Immunocytochemical analysis of brain tissue showed that the transplanted human UCB-derived MSCs could survive and migrate around by the center of transplant site.There were (12.67?2.73)% of MSCs differentiated into astrocyte-like cells.mNSS showed that the score of transplant group was lower than that of control group on the first,7th,14th,21th and 28th day,and the differences of score points between 2 groups on the 14th,21th and 28thday were statistically significant(Pa
6.Effect of OA kneepad on apoptosis genes Bcl-2 and p53 expression in articular cartilage cells of experimental knee osteoarthritis.
Mu-Nan LIN ; Xian-Xiang LIU ; Shui-Liang WANG ; Feng-Hua LAN ; Xi-Hai LI ; Jian-Hua LIU
China Journal of Orthopaedics and Traumatology 2009;22(9):688-691
OBJECTIVETo study the effects of kneepad on expression of Bcl-2 and p53 mRNA of chondrocyte in white rabbits with knee osteoarthritis, so as to explore and treatment mechanism of OA kneepad on apoptosis of chondrocytes of rabbits with knee osteoarthritis in molecular degree.
METHODSForty-four Japanese healthy 6-month-old rabbits (equal male and female,the weight ranging from 2 to 2.2 kg) were used to establish knee osteoarthritis models by modified Hulth method. The rabbits were randomly divided into 6 groups: normal group, model group, control group (microwave), experimental group 1 (electricity), experimental group 2 (thermal), experimental group 3 (kneepad). Ten rabbits in the normal group were breed with conventional method; 9 rabbits in the model group were breed with conventional method after model made; 9 rabbits in the control group were treated with microwave for 30 minutes, one time daily; 9 rabbits in the experimental group 1 were treated with electricity (density wave) for 30 minutes,one time daily;8 rabbits in the experimental group 2 were treated with hot (hot soft membrane) for 30 minutes, one time daily; 9 rabbits in the experiment group 3 were treated with electrothermal (OA knee pad) for 30 minutes, one time daily. All the rabbits were treated for 16 weeks and then sacrificed. The expressions of Bcl-2 and p53 mRNA of chondrocytes in knee joint were detected by using fluorescence quantitative RT-PCR method.
RESULTSAt the 16 hthek,th e OD260/OD280 value range of total RNA extracted from rabbit articular cartilage tissue in each group were all at 1.80 to 2.00,wh ich indicates high RNA purity. The p53 relative mRNA in articular cartilage cells of model group,th e control group,th e experimental group 1 ,r oup 2,gr oup 3 were overexpressed,an d Belc2 mRNA expression levels of articular cartilage cells were low expression,an d compared with the normal group there were significant differences (P < 0.01). Belc2, p53 mRNA expression in articular cartilage cells,th ere were significant differences (P < 0.01) between the control group, experimental group 1, group 2, group 3 and model group. The results between the control group, experimental group 1 ,group 2 and group 3 had significant differences (P < 0.01).
CONCLUSIONOA-kneepad can up-regulate the mRNA expression of Bcl-2 as well as down-regulate the mRNA expression of p53, thereby to inhibit the apoptosis of cartilage cells and delay the degeneration of articular cartilage changes.
Animals ; Apoptosis ; physiology ; Disease Models, Animal ; Female ; Knee Joint ; metabolism ; pathology ; Male ; Osteoarthritis, Knee ; genetics ; pathology ; therapy ; Protective Devices ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics
7.Value of 3D-OCT in the diagnosis of macular disease before cataract surgery
Lin, LIU ; Hua, ZHENG ; Ni, LI ; Ping, TONG ; Yong-Gang, SHUI ; Lan, YU ; Shan, LIU ; Fei, SU ; Xuan-Chu, DUAN
International Eye Science 2017;17(9):1765-1767
AIM:To investigate the value of 3D-optical coherence tomography (OCT) in the diagnosis of macular disease before phacoemulsification.METHODS:Clinical records of 423 cataract patients (512 eyes) who underwent phacoemulsification combined with intraocular lens implantation in our hospital from June to December in 2015 were retrospectively analyzed.In addition to preoperative routine examination of fundus, Topcon 3D-OCT 2000 was used to examine the macula, the detection rate of macular disease was compared, risk factors of cataract combined with macular disease were analyzed.RESULTS:The OCT image results of 305 cases (384 eyes, 72.1%) were successfully obtained, 133 cases showed macular disease (146 eyes), the detection rate was 28.5% (95%CI:27.64%-29.40%);the macular disease of 35 cases (37 eyes) were detected by routine examination of fundus before operation, the detection rate was 7.2% (95%CI:6.72%-7.74%);the detection rate of 3D-OCT was significantly higher than routine examination of fundus for macular disease (χ2=79.05, P<0.01).Female, over 65 years old, surgical history of diseased eye, and high myopia were risk factors of cataract combined with macular disease, the relative risk was 1.705 (95%CI:1.091,2.664), 1.893 (95%CI:1.219,2.939), 6.593 (95%CI:2.027,21.447) and 95%CI:5.130 (2.841,9.263) respectively, the risk of cataract combined with macular disease showed an increasing trend with rising age.CONCLUSION:In preoperative examination of cataract patients, 3D-OCT has higher sensitivity in the detection of macular disease, especially for women, over 65 years old, high myopia and surgical history of diseased eye, 3D-OCT can be used as a routine preoperative examination.
8.Study of Bilingual Teaching for Medical Microbiology
Shui-Xiang XIE ; Xiao-Li WANG ; Wen-Ping ZHANG ; Lian-Lan MA ; You-Tian ZHONG ; Gao-Lu CAO ; Zhen HUANG ;
Microbiology 1992;0(01):-
Bilingual teaching is adapted to the development of higher education in china.Based on actual fact of college,teaching mode,evaluation and effect of bilingual teaching on medical microbiology were studied,which started with necessity of bilingual teaching to use original edition teaching material in English. The result would provide some gist to choice the suitable pattern of bilingual teaching for other subject of our college.
9.Developing Students' Ability to Practice Resulting from Reforme of Experimental Teaching Pattern of Medical Microbiology
You-Tian ZHONG ; Li WANG ; Xiao-Li WANG ; Zhi-Chun LIU ; Shui-Xiang XIE ; Lian-Lan MA ;
Microbiology 1992;0(05):-
To develop Students' Practical Ability according to the teaching requirement and culture aim of preventive medicine major,the teaching plan,teaching content,teaching methods,and experimental check-ing methods were explored and the experimental teaching pattern of medical microbiology adapted to pre-ventive medicine major was constructed.The investigation showed that the experimental teaching pattern helped to cultivate the students' operating ability,thinking of scientific research and ability of aggregate and solving analysis.Moreover,it helped to develop the students' co-operative consciousness and team spirit.It indicated that the new pattern was superior to the traditional experimental teaching.
10.Emergency response of infectious snails detected after interruption of schistosomiasis transmission in Hannan District, Wuhan
ZOU Yu-ting ; XU Ming-xing ; LUO Hua-tang ; SONG Xiu-lan ; CHEN Qiu-qin ; WANG Hao ; ZHOU Shui-mao
China Tropical Medicine 2023;23(2):131-
Abstract: Objective To analyze the emergency response and long-term intervention effects after the detection of infectious snails epidemic by loop-mediated isothermal amplification (LAMP) assays in Hannan District, Wuhan City, and to explore the application of LAMP in early surveillance and early-warning of schistosomiasis transmission. Methods Snails picked up by the risk monitoring system in Hannan District were examined by anatomical microscopy and LAMP technology to identify the schistosomiasis infection. Emergency response and intensive intervention were initiated in the environment where positive snails appeared, and the long-term effects were evaluated. Results In May 2018, the infectious snails were detected by LAMP technology in Hannan District, and the positive snails were located in Zhujiacha, Dongzhuang Village, Obstacles and weeds were removed and buried by machine in Zhujiacha. 12 700 m2 of snails were killed by drugs, and the mortality rate of snails was more than 80%; no new seropositive persons were found in the emergency examination within 500 m of the positive snail sites. 506 people were examined in Dong Zhuang Village at the end of the year, and 30 positive IHA cases were detected with a blood positive rate of 5.93%, no positive fecal test was found, and all positive blood test patients took preventive medication. The monitoring results of sentinel rats and wild feces were all negative. Health education was carried out, 7 warning signs were deployed and refreshed, and 500 publicity brochures were distributed. After nearly three years of intensified intervention and monitoring in the villages where the positive environment is located, and the density of snails on the stubborn snail has dropped from 0.094/frame to 0.027/frame, and the positive rate of blood test in Dongzhuang Village has steadily dropped from 5.93% to 3.74%. Conclusions The infected snails missed by microscopy were detected by LAMP in Hannan District, which created conditions for the rapid emergency treatment of environment and elimination of positive snail and improved the sensitivity of the surveillance and early warning system in transmission-interrupted areas.