3.Detection of p53 and p21~(WAF1) in cholangiocarcinoma
Xin WANG ; Jianxi ZHANG ; Jie CHEN
Chinese Journal of Hepatobiliary Surgery 1998;0(06):-
Objective To clarify the clinical significance of p21 WAF1 and the relationship between it and p53. Methods Samples of human cholangiocarcinoma (HC) tissue and paired normal bile duct tissue adjacent to the tumor from 30 patients with HC were employed in this study. In situ hybridization (ISH) was used to detect the p53 and p21 mRNA expression. Immunohistochemistry (IHC) was applied to analyze p53 gene mutation and P21 protein expression. Results p21 WAF1 And p53 gene protein expression was detected in 36.7% (11/30) and 53.3% (16/30) of the carcinoma specimens, respectively, and none of the paired tissue by IHC. p53 Positivity was related to local lymph node metastasis. Comparing with non-lymph node metastatic group, p53 positivity in metastatic group was significantly higher. p53 Positivity of cholangiocarcinoma in clinical stage III was significantly higher than that in clinical stages I and II. The survival time was significantly shorter in patients with P53 protein expression than in those without. It was found that p53 expression was not associated with p21 WAF1 expression. Conclusions P53 Positivity may be correlated with tumor development but not tumor occurrence. The fact that p53 expression was not associated with p21 WAF1 expression indicates that p53-independent activation of p21 WAF1 may exist.
4.The application of urinary kallidinogenase in recombinant tissue plasminogen activator intravenous thrombolytic treatment in patients with acute cerebral infartion
Jie CHEN ; Xin YAN ; Yuheng SUN
Chinese Journal of Geriatrics 2009;28(8):685-687
ObjectiveTo evaluate the safety and efficacy of urinary kallidinogenase for recombinant tissue-type plasminogen activator (rt-PA) intravenous thrombolytic treatment in patients with acute cerebral infartion MethodsA randomized control study was applied. All 44 patients with acute cerebral infartion were randomized 1:1 to the experimental group (22 cases) and the control group (22 cases). Patients were administrated rt-PA(0. 9 mg/kg)in control group, and patients were given urinary kallidinogenase by intravenous drip (0.15 PNAU/d, for 7 days) after rt-PA intravenous thrombolytic treatment (0.9 mg/kg)in experimental group. The main evaluation index was the incidence of symptomatic intraeerebral hemorrhage within 24 hours, and the secondary assessing items were NIHSS and BI. ResultsThere was 1 case (4.6%) with symptomatic intracerebral hemorrhage in the experimental group and 2 (9.1%) in the control group (X2 =0.00, P= 1.000),and reinfarction rate showed a decreasing tendency in experimental group (18.2% vs. 31.8%, X2=1.091,P=0.296). Compared with the control group, the NIHSS scores were significantly lower 1,21,90 days after thrombolytic therapy (t=2.119, 2.913, 2.187);P=0.041, 0.0 06, 0.042),and the BI scores were obviously higher at 90 days after thrombolytic therapy in experimental group(t= 2.39,P= 0.012). ConclusionsWithout increasing the risk of intracerebral hemorrhage, urinary kallidinogenase may improve the curative effect for rt-PA intravenous thrombolytic treatment in patients with acute cerebral infartion
5.A study of deformable image registration of high-dose-rate brachytherapy CT image to intensity modulated radiation therapy CT image
Xin ZHEN ; Haibin CHEN ; Yang XIAO ; Jie HU ; Linghong ZHOU
Chinese Journal of Radiation Oncology 2015;24(2):209-212
Objective To study a novel method for the high-dose-rate brachytherapy (HDR) CT image to the intensity modulated radiation therapy (IMRT) CT image deformable image registration and dose accumulation.Methods The applicator in the HDR CT image is first segmented and removed,then a deflation step is performed on the applicator-free HDR CT image by solving the Navier-Stokes equation.Demons algorithm is utilized to register the deflated HDR CT image to the IMRT CT image,along with the HDR dose.The deformed HDR dose is then added on the IMRT dose and yield the final accumulated dose.Results The HDR CT image and IMRT CT image,as well as the corresponding dose distribution,from five cervical cancer patients are used for evaluation of the proposed algorithm,the results show that the proposed method can effectively get rid of the influence of the applicator and produce an accurate accumulated dose.Conclusions Dose accumulation and supervision is an important step in adaptive radiotherapy for accurate dose delivery and treatment plan re-optimization.The proposed method in this study can effectively accumulate the HDR dose to the IMRT dose domain,and the accuracy is proved to be sufficient for clinical needs.
6.Studies on chemical constituents from fruits of Morus alba L.
Xin WANG ; Hongqing WANG ; Jie KANG ; Chao LIU ; Ruoyun CHEN
Acta Pharmaceutica Sinica 2014;49(4):504-6
Chemical investigation of fruits of Mours alba L. lead to the isolation of fifteen compounds by various chromatographies such as silica gel, Sephadex LH-20, RP-C18 column chromatography. Their structures were determined to be: 1-[5-(2-formlfuryl) methyl] dihydrogen 2-hydroxypropane-1, 2, 3-tricarboxylate 2, 3-diethyl ester (1), 1-[2-(furan-2-yl)-2-oxoethyl] pyrrolidin-2-one (2), divaricataester A (3), methyl 1-[2-(furan-2-yl)-2-oxoethyl]-5-oxopyrrolidine-2-carboxylate (4), 1-[2-(furan-2-yl)-2-oxoethyl]-5-oxopyrrolidine-2-carboxylic acid (5), L-pyroglutamic acid (6), L-pyroglutamic acid ethyl ester (7), 3-O-caffeoylquinic acid methyl ester (8), 3-O-caffeoylquinic acid ethyl ester (9), 5-O-caffeoylquinic acid methyl ester (10), 5-O-caffeoylquinic acid ethyl ester (11), 4-O-caffeoylquinic acid methyl ester (12), 4-O-caffeoylquinic acid methyl ester (13), 4-O-caffeoylquinic acid (14), 3-O-caffeoylquinic acid (15), respectively, based on the spectral analysis such as NMR, MS etc. Compounds 1-14 were isolated from this genus for the first time, among which 1 was a new compound.
7.Primary Study of Producing UMP by Overexpressing URA 5 and URA 3 Genes in Saccharomyces cerevisiae
Gao-Xin LEI ; Yong CHEN ; Lin XU ; Han-Jie YING ;
China Biotechnology 2006;0(12):-
To increase the biotransfomation efficiency from the orotic acid to the uridine 5'-monophosphate(UMP),URA5 gene encoding orotate phosphoribosytransferase was amplified from Saccharomyces cerevisiae BY4742 by PCR,then it was inserted into the expression vector pYX212(contained orotidine monophosphate decarboxylase gene URA3)and the pYX212-URA5 was transformed into Saccharomyces cerevisiae BJX12 by electroporation.The recombinant strain was elementarily used to convert orotic acid to UMP.The results showed that pYX212-URA5/BJX12 could accumulate 7mmol/L UMP from 32mmol/L orotic acid in 26h,significantly higher than both control groups pYX212/BJX12(2.7mmol/L) and BJX12(2.4 mmol/L).
8.Comparison of two methods testing synergistic action of Reduning and cefoperazone sodium/sulbactam sodium on extensive drug resistant Acinetobacter bauman
Xin LI ; Guifang YANG ; Huayu CHEN ; Jie GENG ; Zhijie JIA
International Journal of Laboratory Medicine 2016;37(6):723-725
Objective To observe the results of broth dilution method and disc diffusion method to test the synergistic effect of Reduning and cefoperazone sodium / sulbactam sodium(SCF) on extensive drug resistant Acinetobacter bauman (XDR-AB) in vitro environment ,and compare their compliance to guide the clinical medication .Methods A total of 12 strains of XDR-AB from infec-tion patients in our hospital in 2015 were collected ,the strain was sub cultured .Firstly ,observe the minimum inhibitory concentra-tion (MIC) of SCF and Reduning on XDR-AB alone and in combination by broth dilution method .And then judge the synergy effects through calculation .Secondly ,the inhibition ring diameter and the synergy effects was detected using the disc diffusion meth-od .Results The MIC of Reduning and SCF in combination on XDR-AB was declined compared with them alone .The Fractional in-hibitory concentration of Reduning and SCF in combination on XDR-AB were equal or less than 0 .5 ,they had synergistic effect on XDR-AB .The inhibition ring diameter of Reduning was 10 mm tested by disk diffusion method .Different strains of XDR-AB on SCF bacteriostatic annulus diameter difference ,5 strains were 15 mm ,3 strains were 16 mm ,and 4 strains were 17 mm .Reduning and SCF appeared synergistic effect according to the inhibition ring diameter expanded when they effected on XDR -AB in combina-tion .Conclusion In vitro ,Reduning combined with SCF on XDR-AB has good synergistic effect .Compared with broth microdilution checkerboard dilution method ,disk diffusion method is more simple and convenient ,but it has a certain subjective on judging re-sults ,which is better to operate by experienced person .
9.Genetic polymorphisms of 19 STR loci in han popu-lation from Hainan and its application in paternity testing
Chunbao CHEN ; Zhen SU ; Xin TIAN ; Hanhua WU ; Jie WANG
Chinese Journal of Forensic Medicine 2017;32(3):276-280
Objective To construct a database for the genetic polymorphism of 19 STR loci in Han population from Hainan province. To investigate the application of 19 STR loci in the paternity testing. Methods The genotypes of 462 unrelated individuals in Hainan were detected with GoldeneyeTM 20A PCR Amplification Kit. 19-STR database was acquired, analyzed and evaluated in 283 paternity testing cases. Results No deviations of allele frequency from Hardy-Weinberg equilibrium expectations were found for Chi-square test (P>0.05). Observed heterozygosity (Hobs) varied between 0.603 and 0.914, total discrimination power (TDP) of 19 STR loci was more than 0.999999999999999, cumulative probability of exclusion (CPE) for triplet cases was 0.999999994. In all 283 paternity testing cases, triplets and duos were 170 and 113 respectively; there were 36 (12.7%) excluded cases comparing to 247 confirmed cases (87.3%). 14 mutation events were observed, and all were one-step mutation. Conclusion 14 out of 19 loci showed highly polymorphic in Han population from Hainan, and 19 STR system has high cumulative probability of exclusion and can meet the needs of paternity test of the local region. But mutation should be paid special attention to.
10.J774A. 1 cell apoptosis induced by Leptospira interrogans lipopolysaccharide and apoptotic regulation of associated signaling pathways
Shijun LI ; Minghuan CHEN ; Xin ZHAO ; Jie YAN
Chinese Journal of Microbiology and Immunology 2010;30(11):1014-1019
Objective To determine the effect of Leptospira interrogans lipopolysaccharide (L-LPS) inducing apoptosis of murine mononuclear-macrophage cell line( J774A. 1 ), and apoptotic regulation of Toll-like receptor(TLR) and associated intracellular signaling pathways. Methods Lipopolysaccharide (L-LPS) of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai 56601 was prepared using phenol-water method. The effects of L-LPS inducing J774A. 1 cell apoptosis and the apoptosis-blocking with FasL neutralizing antibody were detected by flow cytometry. Real-time fluorescent quantitative RT-PCR (qPCR) and flow cytometry were performed to measure the changes of Fas/FasL mRNA and protein expression levels in J774A. 1 cells before and after L-LPS treatment. The regulations in L-LPS-induced cell apoptosis by TLR2 and TLR4 as well as p38MAPK, JNK, ERK pathways were determined by either TLR2 or TLR4 antibody blocking test, signaling pathway blocking test and flow cytometry. Results 56.50%, 69.28% and 24.35% of the J774A. 1 cells after treatment with 100 ng/ml L-LPS for4, 12 and 24 h were apoptotic,while the apoptosis rates were decreased to 11.21%, 21.58% and 12.70% after the cells blocked by FasL neutralizing antibody(P <0.05). The levels of FasL and Fas mRNAs in J774A. 1 cells treated with L-LPS for 4, 12 and 24 h were elevated with 1.34, 2.12, 2.10 times and 2.45, 3.87, 3.12 times compared to those in the L-LPS untreated cells (P < 0. 05 ), respectively, while the expression rates of FasL and Fas proteins were upregulated to 18.61%, 60.13%, 42.75% and 76.34%, 85.70%, 77.92% from 4.82% and 15.32% apoptotic rates in the L-LPS untreated cells, respectively( P <0.05 ). The L-LPS-induced apoptosis rate( 11.54% ) of TLR2 antibody blocked J774A. 1 cells was significantly lower than that(66.56% ) of the J774A. 1 cells without TLR2 antibody blocking( P <0.05 ), but L-LPS-induced apoptosis rate of TLR4 antibody blocked J774A. 1 cells was as high as 55.27% ( P > 0.05 ). Compared to the apoptosis rate (62.17%) in the p38MAPK and JNK pathway-free J774A. 1 cells, the L-LPS-induced apoptosis rates in p38MAPK blocked cells(20.54% ) and JNK blocked cells(47.98% ) were significantly lower( P <0.05 ),and the apoptosis rate in ERK blocked cells was as high as 61.72% ( P > 0.05 ). Conclusion L-LPS was recognized by TLR2 and upregulates both Fas and FasL expression via p38MAPK and JNK pathways, which involving in the process of the L-LPS-induced cell apoptosis.