1.Suppression effect of RNA interference on expression of EGFR of SKOV3 cell line
lan, DAI ; wen, DI ; chuan-wei, DING
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To investigate the suppression effect of RNA interference(RNAi) on the expression of epidermal growth factor receptor(EGFR) of ovarian cancer SKOV3 cell line. Methods SKOV3 cells were transfected with synthetic EGFR sequence-specific dsRNA by lipofectamin 2000.The expression level of EGFR mRNA and protein were detected by RT-PCR and Western blotting,respectively.The proliferation of transfected SKOV3 cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay,respectively. Results The suppression rates on EGFR mRNA and protein by sequence-specific dsRNA-EGFR were 73.65% and 58.94%,respectively.The cell proliferation was inhibited by 22.54%,35.76% and 31.07%,respectively at 24,48 and 72 h after transfection.The adhesion of sequence-specific dsRNA transfected SKOV3 cells for 30 min and 90 min were reduced by 12.11% and 18.66%,respectively.The migration and invasion of SKOV3 cells were decreased by 25.47% and 22.08%,respectively. Conclusion The down-regulation of EGFR expression of ovarian cancer by sequence-specific dsRNA can lead to the suppression of proliferation,adhesion,migration and invasion of ovarian cancer cells.
2.Four types of surgery for the treatment of adenoid hypertrophy
Lide WU ; Xuejun TAN ; Yi WEN ; Xiaoxia WEN ; Peng DAI ; Di WU ; Shaoyun CHEN ; Xiaoping CHEN
Chongqing Medicine 2016;45(27):3808-3810,3813
Objective To explore the effection of four types of surgery for the treatment of adenoid hypertrophy . Methods 148 cases of with adenoid hypertrophy treated in our hospital between April 2012 and April 2015 were chose;they were randomly divided into 4 groups ,each group of 37 people .A group of patients with adenoid hypertrophy were taken traditional ade‐noidectomy curettage;Group B with nasal endoscopic adenoidectomy and cutting aspiration biopsy ;Group C by adenoidectomy shave their + residual endoscopic adenoidectomy bit cut method combined treatment ;Group D with nasal endoscopic adenoidectomy plas‐ma cutting treatment .The curative effect ,operation time ,blood loss were observed ;patients were followed‐up for half a year ,ade‐noidectomy residual rate and complications of each group were compared .Results The total effective rate of B ,C ,D three groups were significantly higher in group A patients (χ2 =7 .731 ,5 .045 ,7 .731 ,P<0 .05) ,the efficient between three groups was not sta‐tistically different (P>0 .05) .B ,C ,D three groups of operation time is significantly higher than A group of patients (t=5 .819 , 5 .829 ,2 .759 ,P<0 .05);B and C group had long operation time than group D (t=3 .555 ,3 .637 ,P<0 .05);But operation time of B and C had no significant difference between the two groups (t=0 .149 ,P>0 .149) .Bleeding of B and C group were significantly higher than group A (t=9 .305 ,4 .126 ,P<0 .05);Group D was significantly lower than A ,B ,C three group (t=8 .054 ,16 .559 , 12 .837 ,P<0 .05);Group C and group B was significantly higher than the bleeding (t=5 .739 ,P<0 .05) .Retention rate of group A is significantly higher than the other three groups (χ2 =31 .308 ,31 .308 ,24 .667 ,P<0 .05) ,the residual rate of B ,C ,B group were lower ,there was no statistically significant difference(P> 0 .05) .Complication rates between the four groups was no statistical difference(P>0 .05) .Conclusion we should choose the right means of surgical treatment according to patients condition and eco‐nomic situation to .
3.Regulation of microRNA-199a on adhesion, migration and invasion ability of human endometrial stromal cells
Lan DAI ; Liying GU ; Jie ZHU ; Jun SHI ; Yao WANG ; Fang JI ; Wen DI
Chinese Journal of Obstetrics and Gynecology 2011;46(11):817-821
Objective To study the regulation of microRNA 199a (miR-199a) on adhesion,migration and invasion ability of human eutopic endometrial stromal cells (ESC) from patients with endometriosis.Methods ESC were transfected with miR-199a mimics or negative control (NC) RNA by lipofectamine 2000.The adhesion,migration and invasion ability of ESC were detected by cell adhesion assay,scratch assay,cell migration assay and matrigel invasion assay,respectively.Luciferase reporter assay was used to evaluate whether IKKβ was the target gene of miR-199a.The expression of ikappa B kinase beta (IKKβ),inhibitory kappa B alpha (IκB-α),phospho-IκB-α (p-IκB-α) and nuclear factor-kappa B (NF κB) protein were measured by western blot.Results ( 1 ) Adhesion potential:the adhesion inhibitory rates were ( 14 ± 4 )% in miR-199a group and 0 in control group,which showed significant difference (P<0.01 ).(2) Migration and invasion:in the scratch assay,ESC transfected with miR-199a exhibited a lower scratch closure rate than that of controls.In migration and invasion assays,the migration and invasion ability of miR-199a group were significantly decreased compared with those of NC group [ 130 ± 31 vs.247±36 (P<0.01); 63 ± 15 vs.133 ± 17 (P<0.01),respectively].(3) The luciferase activity of miR-199a group was significantly lowered than that of control group [ 0.160 ± 0.006 vs.0.383 ± 0.083 ( P <0.01 ) ].The protein levels of IKKβ,p-IκB-α,IκB-α and NF-κB of 0.350 ±0.195,0.443 ±0.076,1.970 ±0.486 and 0.454 ± 0.147 in miR-199a group were significantly different compared with the NC group in which the protein levels were set at 1.000 ( P < 0.01 ).Conclusions miR-199a can inhibit the adhesion,migration and invasion of the ESC.IKKβ is the target gene of miR-199a in ESC.One of the mechanisms of the inhibition effect is probably that miR-199a inhibits the activation of NF-κB signaling pathway by targeting IKKβ gene.
4.Study of local immunity of lower genital tract infections
Dai ZHANG ; Zhaohui LIU ; Qinping LIAO ; Jingmei MA ; Yufeng SUN ; Shangrong FAN ; Lina HU ; Haijun JIA ; Wen DI ; Wen WANG
Chinese Journal of Obstetrics and Gynecology 2009;44(1):13-15
Objective To investigate the profile of local immunity of vagina and the immune defense mechanisms against lower genital tract infections. Methods Vaginal lavage was collected from healthy women and patients of vulvovaginal candidiasis, bacterial vaginosis, Trichomonol vaginitis, human papilloma virus infection(VVC), and chlamydia trachomatis infection. Each group included 60 cases. The level of interleukin (IL) 2,4,5, 13,8 and human defensin 5 (HD5) were detected by enzyme linked immunosorbent assay(ELISA). Results (1) Cytokine of helper T cell 1(Th1): the level of IL-2 between healthy women and VVC/ bacterial vaginosis (BV)/ trichomonol vaginitis (TV)/ chlamydia trachomatis (CT) patients had no significant difference. The IL-2 level(96±33 )×10-3 pg/L of human papilloma virus (HPV) infection patients was significantly higher than that of healthy women( P<0.05). (2)Cytokine of helper T cell 2 (Th2) : the level of IL-4 between healthy women and VVC/CT patients had no significant difference. The level of IL-5 between healthy women and BV patients had no significant difference. The IL-13 level(42±15)×10-3 pg/L of TV patients was significantly higher than that of healthy women (30±29)×10-3 pg/L (P<0.05). The IL-4 level (103±28)×10-3 pg/L of HPV infection patients was significantly higher than that of healthy women (36±22)×10-3 pg/L (P<0. 05 ). (3) IL-8 : the IL-8 level (5.8± 2.7) pg/L of TV infection patients was significantly higher than that of healthy women (2. 6±2.4) pg/L (P<0.05). The level of IL-8 between healthy women and BV patients had no significant difference. (4)HD5:the HD5 level of TV , BV, VVC, HPV and CT infection patients were significantly higher than that of healthy women (P<0.05). Conclusions (1) HD5 plays an important role in the defence of vaginal epithelial cell. (2) Th2 may be more important than Thl in lower genital tract infections.(3) IL-8 plays an important role in extrinsic source infections.
5.Study on vaginal production of human defensins and the correlated pathogenetie factors of vulvovaginal candidiasis
Wen WANG ; Wen DI ; Qinping LIAO ; Zhaohui LIU ; Ning ZHANG ; Huiying ZHANG ; Dai ZHANG ; Li GENG ; Shangrong FAN ; Lina HU
Chinese Journal of Obstetrics and Gynecology 2008;43(7):486-489
Objective To investigate the correlated pathogenetic factors and vaginal local immunity in vulvovaginal candidiasis (VVC). Methods A case control study was conducted to compare VVC group (60 cases) with normal group (60 cases). All of the women filled up the specific questionnaires. Routine examination, pH test and bacterial culture were done on the vaginal discharge. Cytokines of the vaginal lavage were measured by enzyme linked immanosorbent assay. Results (1) Outcomes of the questionnaires: there was no significant difference between the two groups in educational background,knowledge of gynecologic infection, history of gynecologic infection, hygienic habit, sex life, or use of medicine (P >0. 05). The incidence of chronic cervicitis in normal group (43%, 26/60) was higher than in VVC group (22%, 13/60; P <0.05). (2) There was no difference in vaginal pH between the two groups ( P > 0. 05 ). (3) Detection rate of candida albicans by vaginal discharge routine examination was 72% (43/60). (4) The concentrations of interleukin (IL) 2, and IL-4 in vaginal lavage did not show significant difference between the two groups ( P > 0. 05 ), but the concentrations of human defensin 5,human beta-defensin (HBD) 1, and HBD2 in WC group [(0.94±0.44) mg/L, (3.1±0.4) μg/L,(10±6) μg/L] were higher than normal group (P < 0.05). Conclusions VVC is a common vulvovaginitis. There is no significant correlation between the incidence of VVC and educational background,knowledge of gynecologic infection, history of gynecologic infection, hygienic habit, sex life, or use of medicine in the child-bearing period. Human defensin may be closely correlated with the pathogenesis of VVC.
6.One hemodialysis patient with headache, blurred vision, and hypotension induced by pituitary prolactinoma.
Wen-di DAI ; Wen-Hu LIU ; Dong-Liang ZHANG
Chinese Medical Journal 2012;125(15):2787-2789
We reported a rare case of a dialysis patient coincident pituitary prolactinoma with calcification. A 55-year-old woman who had undergone hemodialysis for 8 years was admitted to the nephrology unit because of headache, blurred vision, and hypotension. Physical examination was normal; endocrinological examination demonstrated elevated serum levels of prolactin (> 4240 mIU/L), but other hormonal profiles, such as growth hormon, adrenocorticotropic hormone, thyroid stimulating hormone, free triiodothyronine, free thyroxine, follicle-stimulating hormone and luteinizing hormone, were absolutely or relatively lower. A cranial computed tomography (CT) suggested saddle area a high-density screenage with an anteroposterior diameter of 1.0 cm. A cerebral magnetic resonance scan confirmed the pituitary adenoma accompanied with calcification. Contrast-enhanced T1-weighted images revealed a less enhancing tumor, 14 mm wide round lesion with a high intensity signal. It enlarges the sella turcica, but the optic chiasma is not displaced. We suggest that in the differential diagnosis of any hemodialysis patient with severe headache, hypotension, and visual disturbances, this syndrome should be considered as prompt pituitary adenoma.
Female
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Headache
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parasitology
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Humans
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Middle Aged
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Pituitary Neoplasms
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diagnosis
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physiopathology
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Prolactinoma
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diagnosis
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physiopathology
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Renal Dialysis
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Vision Disorders
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pathology
7.Study on steroidal saponins from Dioscorea zingiberensis and their platelet aggregation activities.
Jing-jing WANG ; Yi-xun LIU ; Di WEN ; He-shui YU ; Li-ping KANG ; Xu PANG ; Zhao YANG ; Bai-ping MA ; Yun-dai CHEN
China Journal of Chinese Materia Medica 2014;39(19):3782-3787
Using the absorbent resin, silica gel and ODS column chromatography as well as semi-preparative HPLC, ten compounds were isolated from 70% ethanol extract of tubers of Dioscorea zingiberensis C. H. Wright, and their structures were elucidated as trigoneoside XIIIa (1), parvifloside (2), trigoneoside IVa (3), deltoside (4), protobioside (5), lilioglycoside k (6), zingiberensis newsaponin I (7), deltonin (8), prosapogenin A of dioscin (9), and trillin (10) on the basis of NMR and MS spectral data analysis. Among these compounds, 1, 3, 5 and 6 were isolated from this plant for the first time. In the screening test on platelet aggregation, compounds 7 and 8 exhibited induction effect on platelet aggregation, while compound 9 exhibited significant inhibitory effect on platelet aggregation in vitro.
Animals
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Dioscorea
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chemistry
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Male
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Mass Spectrometry
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Molecular Structure
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Platelet Aggregation
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drug effects
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Rats
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Rats, Wistar
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Saponins
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chemistry
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pharmacology
8.Diagnosis of thyroid nodules with ultrasound-guided fine needle aspiration: comparison of solid lesion and complex lesion
Wen-bo LI ; Qing-li ZHU ; Yu-xin NG JIA ; Bo ZHANG ; Qing DAI ; Pin GAO ; Di YANG ; Jian SUN ; Xiao-ping XING ; Xiao-lan LIAN
Chinese Journal of Ultrasonography 2011;20(8):680-683
Objective To compare the efficacy of ultrasound-guided fine-needle aspiration(US-FNA)biopsy in diagnosing solid and complex thyroid nodules with different size. Methods One hundred and seventy-five thyroid FNA biopsies were prospectively performed on 168 patients ranging from 4 to 75 years of age. Sixty-three nodules were surgically excised and the others were clinically followed-up. The cytology diagnoses were categorized into four groups: benign, malignant, suspicious and unsatisfactory. Results There was no significant complication in the all 115 solid and 60 complex thyroid lesions and there were 36and 3 malignant nodules respectively in solid and complex thyroid nodules. The nondiagnostic rates of solid and complex nodules were 7% and 8%. The accuracy of US-FNA in diagnosing complex thyroid nodules was comparatively equal to that of in solid thyroid nodules. In solid thyroid nodules, the sensitivity and accuracy in ≤1 cm group were similar to that of in >1 cm group. Conclusions US-FNA was an accurate and reliable method to diagnose thyroid solid and complex lesions.
9.Expression of Smad4 in leukemia cells.
Yan ZHANG ; Xu CAO ; Ming JIANG ; Li-Dai HA ; Bing-Zhao WEN ; Ling LI ; Hui LIU ; Di ZHONG ; Ren-Yong LIN ; Xiao-Mei LU ; Xiao-Hui FENG ; Hao WEN
Journal of Experimental Hematology 2006;14(4):673-676
Loss of transforming growth factor (TGF)-beta signaling has been implicated in malignant transformation of various tissues. Smad4 plays a central role in the signal transduction of TGF-beta. Deletion or mutation of Smad4 has been described in a number of cancers. This study was aimed to investigate a potential role of Smad4 in leukemia including its expression and location in blast cells. The mononuclear cells were separated from bone marrow of leukemia patients. The samples, blast cells of which were more than 90% in mononuclear cells, were selected. The expression and location of Smad4 protein were analyzed by immunohistochemistry methods. The results showed that the Smad4 protein located mainly in nucleus, part of this protein located in cytoplasma, the expressions of Smad4 were not detected in 6 out of 9 ALL patients, in 7 out of 24 AML patients and in 1 out of 2 CML patients; these leukemia patients, in whose cells the expression of Smad4 was not detected, included one L1 and one L3, four L2, one M0, one M1, two M2a, one M3a, one M4b, one M6 and one CML. In conclusion, the Smad4 protein was mainly in nucleus, the deletion or functional change of Smad4 may related with the pathogenesis of human AML.
Humans
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Leukemia, Myeloid, Acute
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genetics
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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genetics
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Signal Transduction
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Smad4 Protein
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biosynthesis
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genetics
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Transforming Growth Factor beta
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biosynthesis
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genetics
10.Study on the relationship between the polymorphisms and secondary structure of tat exon-1 gene and HIV/ AIDS progress in subtype B' and B'/C.
Xiao-xu HAN ; Di DAI ; Yan ZHANG ; Min ZHANG ; Zi-ning ZHANG ; Ying-ying DIAO ; Wen-qing GENG ; Hong SHANG
Chinese Journal of Epidemiology 2006;27(11):968-972
OBJECTIVETo study the polymorphisms and secondary structure of human immunodeficiency virus (HIV-1) tat exon 1 among subtype B' and B'/C HIV-1 infected people in China and to explore the relationship between the polymorphism of tat exon 1 and the disease progression.
METHODS8 subtype B' and 5 B'/C HIV-1 infected patients with slow disease progression were selected from Liaoning, Jilin and Yunnan province. 26 subtype B' and 9 B'/C HIV-1 infected patients with similar sex, age but with typical disease progression were selected. Provirus was extracted from the whole blood. The gene sequences of the Tat exon 1 were amplified by nest-polymerase chain reaction (nest-PCR). Products were purified and sequenced directly. The sequences were aligned, translated, amino acid substitution were analyzed and secondary structures were predicted.
RESULTSMany amino acid substitution could be found in the exon 1 of Tat in HIV-1 subtype B' and B'/C recombinant strain infected persons with different disease progression except A58T,none of them showed definitely relationship with HIV viral load and disease progression. 23N, 31S, 32Y and 46F were subtype-specific substitutions. No characteristic secondary structure of exon 1 of Tat was found.
CONCLUSIONSome of the mutations of tat exon 1 might be related to HIV viral load and disease progression. However, there was no relationship found between the secondary structure of Tat protein and the disease progression.
Acquired Immunodeficiency Syndrome ; genetics ; pathology ; Amino Acid Substitution ; Disease Progression ; Exons ; genetics ; Genes, tat ; genetics ; HIV Infections ; genetics ; pathology ; Human Immunodeficiency Virus Proteins ; genetics ; Humans ; Polymorphism, Genetic ; Viral Load