1.Effect of ULF gene silence on etoposide induced H1299 cell apoptosis
Wei LIU ; Xin XU ; Xiaoci CAO ; Fei YUAN ; Huiying FANG
Chinese Journal of Biochemical Pharmaceutics 2014;37(4):59-62
Objective To explore the effects of knocking down ULF gene on the apoptosis of non-small-cell carcinoma H1299 cell after treatment with etoposide.Methods Three ULF small interfering RNA(siRNA)sequences and one negative control siRNA sequence were designed and synthesized, and then individually transfected into H1299 cell via lentivirus.The interference efficiency of ULF-siRNA were screened by real-time PCR and Western blotting.Then the most target siRNA was used for apoptosis assay after treatment with etoposide,MTT assay for H1299 cell proliferation,flow cytometry for cell cycle distribution. Results The expression of ULF gene and its protein ULF were down-regulated in H1299 cell when transfected with ULF-siRNA,and ULF-siRNA-1 was the most effective one,which had the highest inhibition rate(80%)of ULF expression.Compared with negative control group,ULF-siRNA group showed an obvious apoptosis after treatment with etoposide,and the inhibition rate of was higher than control group,which was positively correlated with etoposide dose,the difference was statistically significant(P<0.05 ).Flow cytometry showed that compared with the control group,G0/G1 cell cycle in ULF-siRNA group was increased,and S phase cells was decreased,the differences were all statistically significant(P<0.05).Conclusion Down-regulation ULF protein expression through treatment with etoposide can induce apoptosis of non-small-cell carcinoma H1299 cells,and inhibit cell proliferation,which lead to cell cycle arrest.ULF gene may become the new target of gene therapy for cancer.
2.Application value of three-dimensional visualization technology in the resectability assessment and surgical planning for huge hepatic carcinoma
Wei CAI ; Fei XIANG ; Yaohuan HUANG ; Yingfang FAN ; Chihua FANG
Chinese Journal of Digestive Surgery 2017;16(1):53-58
Objective To investigate the application value of three-dimensional (3D) visualization technology in the resectability assessment and surgical planning for huge hepatic carcinoma.Methods The retrospective cross-sectional study was conducted.The clinical data of 48 patients with huge hepatic carcinoma who were admitted to the Zhujiang Hospital of Southern Medical University between January 2012 and June 2015 were collected.The preoperative image of computed tomography (CT) was converted to 3D reconstruction,visual observations and simulated surgery for assessing the tumor resectability through MI-3DVS,and corresponding treatments were performed according to the results of assessment.Observation indicators:(1) 3D reconstruction situations;(2) tumor resectability assessment through simulated surgery:tumor diameter,tumor volume,preoperative standard liver volume (SLV),tumor-free liver volume after simulated resection,future liver remnant (FLR) after simulated resection,hepatic resection rate (HRR);(3) surgical and postoperative situations:surgical procedures,resection extent,operation time,volume of intraoperative blood loss,complications,duration of postoperative hospital stay;(4) typical case analysis;(5) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect the survival of patients and tumor recurrence or metastasis up to June 2016.Measurement data with normal distribution were represented as(-x) ± s.Results (1) Three-D reconstruction situations:48 patients with huge hepatic carcinoma received successful 3D reconstruction and visual observations.Portal vein branches and hepatic vein branches reached level 4 through 3D reconstruction,and spacial position relationship between tumor and intrahepatic vascular backbones or branches can be clearly observed,as well as location and degree of vascular compression and invasion.(2) Tumor resectability assessment through simulated surgery:of 48 patients receiving simulated hepatectomy,26 underwent hepatectomy and 22 didn't undergo hepatectomy based on the assessment of resectability.Tumor diameter,tumor volume,preoperative SLV,tumor-free liver volume after simulated resection,FLR after simulated resection and HRR through assessment of 3D reconstruction and simulated surgery were (12.3-± 2.0) cm,(838 ± 284) mL,(1 884 ± 391) mL,(494 ± 140) mL,(551 ± 184) mL,46% ± 12% in 26 patients with resectable tumor and (14.0 ± 2.0) cm,(1 877 ± 1 240) mL,(2 945 ± 1 194) mL,(666 ± 206) mL,(402 ± 86) mL,62% ± 9% in 22 patients with unresectable tumor,respectively.(3) Surgical and postoperative situations:26 patients with resectable tumor underwent hepatectomy,without occurrence of death.Of 26 patients,21 underwent anatomic hepatectomy,including 12 undergoing right hemihepatectomy,3 undergoing left hemihepatectomy,2 undergoing right lobectomy of the liver,2 undergoing right posterior lobectomy of the liver,1 undergoing left lobectomy of the liver and 1 undergoing resection of hepatic segment Ⅴ + Ⅵ.And 5 underwent non-anatomic hepatectomy,including 2 with reduced right hemihepatectomy,1 with resection of hepatic segment Ⅱ + Ⅲ and partial segment Ⅳ,1 with resection of hepatic segment Ⅵ + Ⅶ and partial segment Ⅴ and 1 with resection of hepatic segment Ⅴ + Ⅵ and partial segment Ⅶ.Operation time and volume of intraoperative blood loss in 26 patients were respectively (6.4 ± 1.3) hours and (712 ±633)mL.Three patients with postoperative pleural effusion and 1 with postoperative bile leakage were cured by symptomatic treatment,without the occurrence of hepatic dysfunction.Duration of postoperative hospital stay was (19 ± 8) days.Of 22 patients with unresectable tumor,14 underwent transcatheter hepatic arterial chemoembolization (TACE),4 underwent portal vein ligation,1 underwent portal vein embolization and 3 abandoned treatment.(4) Typical case analysis:results of 3D reconstruction through MI-3DVS showed that patients underwent portal vein right anterior branch-preserving expanded right posterior lobectomy of the liver,with a smooth recovery.Patients were followed up for 14.0 months,with a good survival and without tumor recurrence and metastasis.(5) Follow-up:40 of 48 patients were followed up for 6.0-33.0 months with a median time of 13.0 months,including 26 with surgery and 14 without surgery.During the follow-up,the median survival time of patients with and without surgery was 20.0 months and 10.5 months,respectively.Twelve patients with surgery had tumor recurrence and metastasis.Conclusion Three-dimensional visualization technology is safe and feasible in the resectability assessment and surgical planning for huge hepatic carcinoma,and it will benefit to reduce risk of surgery.
3.A novel HIF-1 inhibitor--manassantin A derivative LXY6099 inhibits tumor growth.
Fang-Fang LAI ; Xiao-Yu LIU ; Fei NIU ; Li-Wei LANG ; Ping XIE ; Xiao-Guang CHEN
Acta Pharmaceutica Sinica 2014;49(5):622-626
Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.
Animals
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Antineoplastic Agents
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pharmacology
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Breast Neoplasms
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metabolism
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Cell Line, Tumor
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Gene Expression Regulation, Neoplastic
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Heterografts
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Humans
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Hypoxia-Inducible Factor 1
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metabolism
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Lignans
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pharmacology
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Mice, Nude
4.The value of 3.0T MR DWI in pathological grading and staging for cervical carcinoma
Shuhua WEI ; Naiyu LI ; Chao WEI ; Fei GAO ; Xin FANG ; Chuanbin WANG
Journal of Practical Radiology 2017;33(7):1062-1064,1072
Objective To explore the correlation between apparent diffusion coefficient (ADC) and pathological grading and staging of cervical carcinoma.Methods Among 311 patients diagnosed with squamous cell carcinoma,87 patients were met the criteria for inclusion.All the patients were performed routine MR and diffusion-weighted imaging(DWI) with b=1 000 s/mm2 before treatments,and recorded the pathological grading and staging.The ADC values of the lesions were measured on GE AW4.5 workstation,and were compared among the groups with different pathological grading and staging by using LSD and paired t-test.Results The mean ADC value was (0.77±0.079)×10-3 mm2/s in the poorly-differentiated group,(0.88±0.10)×10-3 mm2/s in the moderately-differentiated group,and (1.05±0.084)×10-3 mm2/s in the well-differentiated group.There were significant differences among the different groups (P<0.05).The mean ADC value was (0.84±0.12)×10-3 mm2/s in the stage Ⅰb group, and (0.78±0.12)×10-3 mm2/s in the stage Ⅱa group.The difference was not statistical significance (P>0.05).Conclusion The increasing of the ADC values is correlated with the high levels of tumor cell differentiation, and not correlated with the pathological stages in cervical carcinoma.
5.Differentiation of mesenchymal stem cell in the microenviroment of retinitis pigmentosa
Dong-Mei, HUO ; Fang-Tian, DONG ; Wei-Hong, YU ; Fei, GAO
International Eye Science 2010;10(10):1851-1854
AIM: To assess the differentiation of rat mesenchymal stem cell (MSC) in the microenvironment of retinitis pigmentosa(RP) induced by the administration of sodium iodate. METHODS: In vitro cultured Lewis rat MSC were injected into the subretinal space of NaIO3 induced RP rat eyes (30g/L NaIO3 100mg/kg). To observe the trace and differentiation of MSC by immuno-fluorescent method successively in 5 weeks after the surgery.RESULTS: The majority of the transplanted cells stay in retinal pigment epithelium(RPE) layer and cones and rods layer. From the 2nd week after transplantation, the engrafted MSC expressed PCK and rhodopsin under fluorescent microscope.CONCLUSION: MSC can survive mainly in the outer layer of retina in the microenvironment of RP and differentiate forward the RPE cell and photoreceptor.
6.Expression of C2A domain of synaptotagmin Ⅰ fusion protein and its imaging in the ischemia-reperfusion rat model
Kang-yun, SUN ; Jun-dong, ZHOU ; Fei, SHEN ; Wei, FANG ; Jin-chang, WU
Chinese Journal of Nuclear Medicine 2010;30(5):312-315
Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P <0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.
7.In vitro Effect of Combined Traditional Chinese Medicine (Changqing Capsule) on The Tachyzoites of Toxoplasma gondii
Wei ZHANG ; Furong FANG ; Yuanjiao LIU ; Liandi YANG ; Ruoyu LUO ; Fei GONG ; Hui LU ; Xiaoxia XU
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(01):-
Objective To detect the in vitro effect of the traditional Chinese medicine on the tachyzoites of Toxoplasma gondii. Methods Supernatant (1^5 ml) of different doses of the traditional Chinese medicine (Changqing capsule) was collected by normal saline immersion and 2^5?10+4 Toxoplasma gondii tachyzoites were added in each paste well for 8 hours. Spiramycin, pyrimethamine and azithromycin in different doses were used as controls. Normal saline was used as negative control. Mice were inoculated with drug-treated tachyzoites intraperitoneally or intragastrically. The normal mice were subcultured after 8 days for 3 generations. Results The incident number of the infected mice was significantly different among groups with different drugs and doses: 2/60, 16/60, 10/60 and 10/60 in the groups of Changqing capsule, spiramycin, pyrimethamine and azithromycin respectively (P
8.Investigation of the initial symptom, onset time and predisposing factors of acute cerebral haemorrhage
Fei CAO ; Fang LUO ; Bo HU ; Guirong WEI ; Hui XU ; Jingjie CAI
Chinese Journal of Geriatrics 2008;27(11):824-826
ObjectiveTo investigate the initial symptoms, onset time and predisposing factors of acute cerebral haemorrhage in order to more effectively prevent cerebral haemorrhage. Methods 728 cases with acute cerebral haemorrhage were collected in recent 8 years. The initial symptoms,onset time and predisposing factors of all the patients were investigated. ResultsAcute cerebral haemorrhage in 582 caces (79.9%) occurred at time between 6am and 18pm and 146 cases (20%) at night. The main predisposing causes of acute cerebral haemorrhage were the playing of chess, cards and mahjong (16.1%), quarrels (12.8%), overwork (11.7%), alcoholism (7.2%) and so on. The most common initial symptom was hemianesthesia, accounting for 16.2%, others were vertigo (13.0%), headache ( 11.4%), amaurosis fugax ( 9.3%), speech ambiguity ( 8.6%) and transitory visual disturbance (7.9%).ConclusionsThe preventive measures of cerebral haemorrhage should be focused on daytime, especially in the morning. Sixteen locations where the cerebral haemorrhage is prone to happen should be paid more attention to by the patients with hypertension and the medical staff. Thirteen symptoms,such as hemianesthesia and vertigo,are the aurae of cerebral haemorrhage.
9.Clinicopathologic analysis of four cases of desmoplastic melanocytic nevus
Qian ZHANG ; Jing SU ; Jianying LIU ; Fei GAO ; Fang MEI ; Qian ZHANG ; Mengmeng YE ; Wei ZHANG
Chinese Journal of Dermatology 2017;50(7):503-507
Objective To investigate pathological characteristics and differential diagnosis of desmoplastic melanocytic nevus.Methods Four cases of desmoplastic melanocytic nevus were analyzed based on the clinical manifestations and histological,immunohistochemical and fluorescence in situ hybridization (FISH) features.Results Of the 4 cases,2 were male and 2 were female.Their age ranged from 19 to 30 years with the average age being 26.5 years.The skin lesions were located on the extremities in 3 cases,on the vulva in 1 case.Histologically,the lesions were bilaterally symmetrical intradermal nevus.Nevus cells appeared epithelioid and/or fusiform,some were clustered or scattered in the proliferative fibrous tissue.None of lymphocyte aggregation,necrosis or ulceration was observed.Immunohistochemical examination showed positive staining for S100 and Melan A in 3 cases,positive staining for P16 in 2 cases,Ki-67-1abeling index less than 5% in all the 4 cases,and negative staining for factor XⅢ (FXⅢ) and CD34 in 2 cases.FISH assay showed no copy-number variations in gene loci 6p25 (RREB1),6q23 (MYB),6p11.1-q11.1 (Cep6) and 11q13 (CCND1) in desmoplastic nelanocytic nevus.Conclusion Desmoplastic melanocytic nevus is a kind of histologically unique,benign melanocytic nevus,and immunohistochemical staining for Ki-67,S-100,Melan A and FXmand FISH assay on melanoma can be helpful for the differential diagnosis between cutaneous fibrous histiocytoma and melanoma.
10.Study on the infectious risk model of AIDS among men who have sex with men in Guangzhou
Pei HU ; Fei ZHONG ; Wei-Bin CHENG ; Hui-Fang XU ; Li LING
Chinese Journal of Epidemiology 2012;33(7):667-671
Objective To develop a human immune-deficiency virus (HIV) infection risk appraisal model suitable for men who has sex with men (MSM) in Guangzhou,and to provide tools for follow-up the outcomes on health education and behavior intervention.Methods A cros-sectional study was conducted in Guangzhou from 2008 to 2010.Based on the HIV surveillance data,the main risk factors of HIV infection among MSM were screened by means of logistic regression.Degree on relative risk was transformed into risk scores by adopting the statistics models.Individual risk scores,group risk scores and individual infection risk in comparison with usual MSM groups could then be calculated according to the rate of exposure on those risk factors appeared in data from the surveillance programs.Results Risk factors related to HIV infection among MSM and the quantitative assessment standard (risk scores and risk scores table of population groups) for those factors were set up by multiple logistic regression,including age,location of registered residence,monthly income,major location for finding their sexual partners,HIV testing in the past year,age when having the first sexual intercourse,rate of condom use in the past six months,symptoms related to sexually transmitted diseases (STDs) and syphilis in particular.The average risk score of population was 6.06,with risk scores for HIV positive and negative as 3.10 and 18.08 respectively (P< 0.001 ).The rates of H IV infection for different score groups were 0.9%,2.0%,7.0%,14.4% and 33.3%,respectively.The sensitivity and specificity on the prediction of scores were 54.4% and 75.4% respectively,with the accuracy rate as 74.2%.Conclusion HIV infection risk model could be used to quantify and classify the individual' s infectious status and related factors among MSM more directly and effectively,so as to help the individuals to identify their high-risk behaviors as well as lifestyles.We felt that it could also serve as an important tool used for personalized HIV health education and behavior intervention programs.