1.Effects of specific interfering TACC3 gene expression on proliferation and apoptosis of CD133+CD44+ oral squamous cell carcinoma cells
Rui DUAN ; Yongsheng LI ; Yichao XIA
Chinese Journal of Tissue Engineering Research 2017;21(25):3995-4000
BACKGROUND: Studies have indicated that the abnormal expression of TACC3 is closely related to the occurrence and development of many kinds of tumors, and the expression of TACC3 is up-regulated in these tumors. Therefore, in vitro specific inhibition of TACC3 expression may become an important target for the treatment or intervention of tumor growth.OBJECTIVE: To investigate the mechanism by which TACC3 gene expression regulates cell proliferation and apoptosis in oral squamous cell carcinoma.METHODS: CD133+CD44+ oral squamous cell carcinoma cells were sorted from human oral squamous cell carcinoma cell line Cal-27 by immunomagnetic beads. In experimental group, the shRNA sequence of TACC3 was designed and synthesized, which was then trasnfected into CD133+CD44+ oral cancer stem cells by LipofectamineTM 2000. Empty vector-trasnfected (negative control) and untransfected cells were used as callsed. Forty-eight hours after the transfection, effects of TACC3 gene silencing on proliferation and apoptosis in vitro in CD133+CD44+ oral squamous cell carcinoma were detected by MTT, clone formation test, and TUNEL assay. Western blot assay was used to detect the effect of TACC3 gene silencing on Ki67, Bax and Bcl-2 protein expression in CD133+CD44+ oral squamous cell carcinoma.RESULTS AND CONCLUSION: (1) Cell proliferation. The proliferation rate and expression level of Ki67 were significantly lower in the experimental group than the negative control and untransfected groups (P < 0.05). (2) Clone formation. The clone formation ability in the experimental group was significantly lower than that in the negative control and untransfected groups (P < 0.05). (3) Cell apoptosis. TACC3 gene silencing caused an obvious decrease in Bcl-2 protein expression and a significant increase in Bax protein expression. These findings further confirmed that specific interference of TACC3 gene expression could inhibit the proliferation of CD133+CD44+ cells and promote the apoptosis.
2.Research on Effective Parts of Xiong-Shao Decoction on Protecting Liver, Reducing Enzyme Activity and Oxidative Damage
Xia LI ; Jiuzhang MEN ; Rui LIANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):774-779
This article was aimed to study the therapeutic material basis of Xiong-Shao (XS) decoction on hepatic fi-brosis (HF), and screen effective parts from XS decoction for protecting liver, reducing enzyme activity and oxidative damage. Male wistar rats were randomly divided into the normal group, model group, Fu-Zheng Hua-Y u (FZHY) group, XS group, polysaccharide group, total alkaloids group and the total glycosides group. HF rat model was estab-lished with the intraperitoneal injection of DMN. After modeling, FZHY solution (0.105 g·mL-1), XS decoction (1.610 g·mL-1), crude polysaccharides extract of XS decoction (35.420 mg·mL-1), total glycosides extract solution (25.725 mg·mL-1), and total alkaloids extract of XS decoction (0.196 mg·mL-1) were administered to corresponding treatment group by gavage once a day for 4 weeks, respectively. Rats of the normal group and model group were given equiva-lent normal saline by gavage once a day for 4 weeks. After 4-week drug administration, rats were killed to remove the liver. Automatic biochemical analyzer was used in the detection of serum parameters of liver function, including ALT, AST, TIBL and ALB. Serum SOD activity was detected by xanthine oxidase. GSH-PX activity was tested by DTNB reduction. Serum contents of MDA were measured by TBA. Pathological changes of the liver were observed with HE staining and masson staining. The results showed that compared with the model group, there was no signifi-cant differences between the total alkaloids group and the model group, but levels of serum ALT, AST and TBIL of other treatment groups were significantly decreased, and the serum ALB level was significantly elevated (P< 0.05 or P < 0.01). Compared with the model group, levels of serum SOD and GSH-PX of the FZHY group, XS group and total alkaloids group were significantly elevated (P < 0.01), and level of serum MDA was significantly reduced (P <0.05 or P< 0.01). Comparison among the polysaccharides group, total glycosides group, and model group showed no significant differences. It was concluded that crude polysaccharide and total glycosides fractions were effective parts of XS decoction for protecting liver and reducing enzyme activity. And total alkaloids fraction was the effective part for reducing oxidative damage.
3.Advances of programmed death ligand 1 and its inhibitor in non-small cell lung cancer
Journal of International Oncology 2015;(8):628-630
Programmed death-ligand 1 (PD-L1 ),which is highly expressed in NSCLC,can be divided into two categories:membrane PD-L1 and soluble PD-L1 .PD-L1 participates in tumor immune escape through combining with its receptor.PD-L1 immune checkpoint inhibitors have entered the phase Ⅰ studies and showed a good application prospect.It is also found that the sensitivity of PD-L1 immune checkpoint inhibitors is strongly associated with the expression of PD-L1 in tumor.Therefore,PD-L1 can be used as a biomarker to pre-dict its curative effect.
4.Influence of different hemostatic way on ovarian function in laparoscopic excision of ovarian cysts
Xia WANG ; Cuiying LI ; Rui WANG
Journal of Regional Anatomy and Operative Surgery 2015;(1):87-89
Objective To investigate the influence of different hemostatic way on ovarian function in laparoscopic excision of ovarian cysts. Methods 87 cases with unilateral ovarian cysts by laparoscopic excision were chosen from Jun. 2012 to Jun. 2013 in our hospital, and they were divided into the electrocoagulation group (44 cases) and the suture group (43 cases) according to hemostatic way. The electro-coagulation group were given bipolar coagulation while the suture group were given endoscopic hemostasis. The ovarian hormone levels and blood parameters before and after surgery were compared between the two groups. Results There was no statistically significant differenc in parameters of E2,FSH,LH,P before surgery between the two groups (P>0. 05), and there was no statistically significant differenc before and after surgery in the same group (P>0. 05). Compared with the suture group, there were statistically significant difference in terms of pa-rameters of E2,FSH in the electrocoagulation group(P<0. 05). There was no statistically significant difference in ovary sectional area and PSV between the two groups before surgery (P>0. 05). It was of no statistically significant difference in ovary sectional area between the two groups after surgery (P>0. 05), but it was of statistically significance in PSV (P<0. 05). Conclusion Compared with electrocoagulation, the laparoscopic excision of ovarian cysts with endoscopic hemostasis had less damage to ovarian, and it could protect the ovarian function.
5.Inhibition of Proto-oncogene Pokemon Expression by siRNA Expression Vector in SW480 Cells
Rui GUO ; Xi-Xia LI ; Jun XIE ;
China Biotechnology 2006;0(06):-
To investigate the inhibitory effects of siRNA expression vector on proto-oncogene Pokemon expression in SW480 cells and to provide experimental basis for further research about the biological function of Pokemon.siRNA expression vectors were constructed to express a short hairpin RNA against Pokemon.The recombinants were transfected into SW480 cells with liposome.Cellular morphology and transfection efficiency were observed.The expression of Pokemon were checked by real-time fluorescence quantitative PCR and Western blot.MTT assay was used to detect the effect of siRNA on the growth of SW480 cells and Fluorescence Activated Cell Sorter was applied to study cell apoptosis of SW480 cells.The transfection efficiency was about 36% and the cellular morphology changed greatly at 48h after transfection.siRNA expression vectors could specifically reduce the expression of Pokemon mRNA and protein in SW480 cells.Compared with negative control,the inhibition ratio of Pokemon mRNA expression was 34.2% and 67.7%,in 24th hour and 48th hour,the inhibition ratio of Pokemon protein was 48.3% and 73.6%,in 48th and 72th hour,respectively.Transfection with Pokemon siRNA expression vectors could inhibit cell growth and induce cell apoptosis of SW480 cells.siRNA expression vectors were successfully established that could effectively inhibit the expression of Pokemon in SW480 cells.Pokemon gene silencing could decrease growth and increase apoptosis of SW480 cells.
6.Effects of prenatal pulsed electromagnetic fields on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspring
Xia LI ; Rui CHEN ; Ning JIA ; Hui LI ; Zhongliang ZHU
Chinese Journal of Tissue Engineering Research 2010;14(40):7588-7592
BACKGROUND:Electromagnetic fields can cause changes of the body,especially the nervous system.Effect of pulsed electromagnetic fields(PEMFs)on neural stem cells has been detected.OBJECTIVE:To investigate the effect of prenatal pulsed electromagnetic fields(PPEMFs)on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspdng.METHODS:Sprague Dawley female rats weighing 240-260 g were included and randomly divided into two groups:control and PPEMFs.Rats from the control group were given no interventions.Rats from the PPEMFs group were given PEMFs stress at gestational days 14-20.Each stress was given three times daily for 10 minutes.The male and female offspring rats were sacrificed at 1 month of age and their brains were sectioned to determine the expression of nestin protein and Brdu-positive cells in the hippocampus by immunohistochemistry.RESULTS:The expression of nestin-and Brdu-positive cells in the hippocampus of female and male PEMFS offspring were significantly higher compared with the control group(P<0.001),and there was a significant difference between female and male offspring(P<0.001).The nestin-and Brdu-positive cells in female offspring outnumbered those in male offspring(P<0.001);however,there was no significant difference between female and male offspring in the control group(P>0.05).CONCLUSION:PPEMFs can increase the number and proliferative capability of the neural stem cells in offspring.It may be a pdmary stage of the cascade reaction of the body to the brain damage caused by PPEMFs stress.
7.The relationship among IL-10, TNF gene polymorphisms, Helicobacter pylori infection and gastroduodenal diseases in Hubei Han ethnic
Hui XIAO ; Chun LI ; Yi JIANG ; Rui LI ; Bing XIA
Chinese Journal of Internal Medicine 2009;48(7):552-556
Objective To study the distribution of IL-10 and TNF gene polymorphisms in patients with gastroduodenal diseases in Hubei Han ethnic and their association with Helicobacter pylori (Hp) infection. Methods Six hundred and five patients with gastroduedenal diseases (196 chronic gastritis, 189 gastroduodenal ulcer and 220 gastric cancer) as well as 624 healthy controls were genotyped with PCR-RFLP method for IL-10-1082,-819,-592 and TNFα-308, lymphotoxin-α (LTα) Nco Ⅰ and AspH Ⅰ gene polymorphisms. Hp infection status was determined with ELLS& Results (1) There was significant difference of IL-10-1082 AG + GG genotype among the gastric cancer group with the non-malignant gastric diseases groups and healthy control group (P <0. 05). There was no significant difference of IL-10-592 and -819 gene polymorphisms among gastric cancer patients,non-malignant gastric disease patients and healthy controls (P>0. 05). The genotype frequencies of IL-10-819 were the same as those of IL-10-592. (2) Frequency of IL-10-1082 AG + GG genotype in gastric cancer patients with positive Hp was significantly higher than that in the other three groups (P < 0. 05). (3) Frequency of LTα Nco I AG genotype in gastric cancer patients with Hp infection was signiilcandy higher than that in Hp positive healthy controls (P < 0. 05). There were no other associations between TNFα-308, LTα Nco Ⅰ and AspH Ⅰ gene polymorphisms and Hp infection in gastroduodenal diseases. Conclusions (1) Allele AG + GG of IL-10-1082 was associated with gastric cancer in Han nationality of Hubei province. (2) IL-10-1082 AG + GG,LTct Nco ⅠAG heterozygous genotype may be associated with Hp infection in patients with gastric cancer in Han nationality of Hubei province.
8.Plexiform nerve sheath tumor arising on conjunctiva surface: report of a case.
Rui-nan XIA ; Lan-fen YANG ; Shu-hua LI
Chinese Journal of Pathology 2005;34(12):825-826
Conjunctiva
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surgery
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Diagnosis, Differential
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Eye Neoplasms
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metabolism
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pathology
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surgery
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Humans
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Infant
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Male
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Neoplasm Recurrence, Local
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Nerve Sheath Neoplasms
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metabolism
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pathology
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surgery
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Reoperation
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S100 Proteins
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metabolism
9.The effects of Gefarnate in treatment of rat model of colitis induced by trinitrobenzene sulphonic acid
Pingfei TANG ; Jin LI ; Wencheng HE ; Rui ZHOU ; Bing XIA
Chinese Journal of Digestion 2009;29(9):554-558
Objective To investigate the effects of Gefarnate on expression of myeloperoxidase (MPO),cyelooxygenase-1 (COX-1) and COX-2 in trinitrobenzene sulphonic acid (TNBS) induced experimental colitis in rats and its therapeutic effects on ulcerative colitis. Methods Forty female Sprague-Dawley (SD) rats were randomly divided into 4 groups with 10 each. The rats in group A, B and C were infused with TNBS/alcohol by enema. After the production of colitis, the rats in group A or B were treated daily with 1 ml of normal saline or with 1 ml of 5-ASA (100 mg/kg) by enema,and those in group C were treated daily with 1 ml of Gefarnate by gavage. Group D was served as normal control. After the production of colitis,animals were sacrificed at day 7 and 14 with 5 in each group. The macroscopic changes of the colon were evaluated according to disease activity index (DAD scoring and histological change was assessed by HE staining. MPO activity of the mucosa was detected by biochemical methods. Expressions of COX-1 and COX-2 in tissues were detected by immunohistochemistry. Results Compared with group A, macroscopic and histological scores and MPO activity were significantly decreased in group B and C (P<0.05). The expressions of COX-1 at day 7 and 14 were 1.86±0.51 and 1.96±0.41 in group B, 1.73±0.68 and 1.79±0.6 in group C, 1.91±0.34 and 1.99±0.45 in group D, respectively, which were significantly higher than those in group A (0.87±0.18 and 0.93±0.15, P<0.05). Whereas the expressions of COX-2 at day 7 and 14 were 1.53±0.19 and 0.73±0.15 in group B, 1.73±0.94 and 0.86±0.29 in group C, 0.24±0.18 and 0.18±0. 16 in group D, respectivley, which were significantly lower that those in group A (3.50±0.2;3 and 3.06±0.27). There was a significant difference between group D and group B or C (P<0.05). Conclusions Gefarnate provides a therapeutic effect during TNBS-induced colitis in rats, which is similar to that of 5-ASA. The mechanisms are involved in decreasing the concentration of colonic MPO and regulating the expression of COX-1/COX-2.
10.Quantitative assessment of iron load in myocardial overload rabbit model: preliminary study of MRI T2* map
Lu HUANG ; Rui HAN ; Zhiwei LI ; Sishu YUAN ; Liming XIA
Chinese Journal of Radiology 2014;48(3):236-240
Objective To preliminarily investigate the feasibility of MRI-T2* map in evaluating myocardial iron load of myocardial iron overload rabbit models.Methods Eleven rabbits were included in this study and divided into two groups,myocardial iron overload group (n =10) and the control group (n =1).Iron dextrin (dose of 50 mg/kg) was injected in muscles of thigh once a week,totally 12 weeks.Serum iron test and MRI examination were performed before iron injection,and 1 week to 12 weeks after iron injection.MRI scan protocol included short axial T2* map of the left ventricle and cross-section T2* map of the liver.T2* and R2* of the heart and the liver were measured.One rabbit was killed after MRI examination at pre-iron injection,1 week to 8 weeks,11 weeks and 12 weeks after iron injection,respectively.Heart and liver were avulsed to undergo in vitro MRI scan and then paraffin embedded for pathological slices.MRI scan protocol and measurements of the heart and the liver samples were the same to that of in vivo ones.Pearson correlation was used to calculate the relationships between the parameters.Results Myocardial T2* [(32.5 ± 8.3 ms)] and R2* values [(38.4 ± 7.9) Hz] had significant correlation with injecting iron content(1 033.2 ± 673.4 mg),the Pearson coefficients were-0.799 (P =0.001) and 0.770 (P =0.002),respectively.Myocardial T2 had no significant correlation with liver T2* values (r =0.556,P =0.070).T2* values of heart and liver in vivo [(32.5 ± 8.3) ms and (8.8 ± 5.4) ms],respectively had strong correlation with those in vitro [(19.4 ± 6.5) ms and (9.8 ± 5.0) ms],respectively (r =0.757,P =0.007 and r=0.861,P=0.001).T2* and R2* values of the heart and the liver in vivo and in vitro had no significant correlations with serum iron (P > 0.05).On Prussian blue staining slices,blue particles of myocardium,sinus hepaticus and hepatocyte increased with injecting iron content.Conclusions It is feasible for MRI-T2* map to evaluate the myocardial iron load noninvasively.It may provide reliable information for detecting myocardial iron overload in patients with iron overload at an early stage.