2.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
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.
5.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).
6.The MRI diagnosis of lung cancer intramedullary spinal cord metastasis
Jie ZOU ; Yingjie ZHAO ; Xin PAN ; Liying CHEN
Chinese Journal of Radiology 2001;0(02):-
Objective To discuss the MRI features and pathogenic mechanisms of lung cancer intramedullary spinal cord metastasis (ISCM). Methods Four cases with clinically and 3 cases pathologically proved lung cancer ISCM were analyzed retrospectively. Turbo spin-echo sequence T 1 and T 2 weighted images were acquired in all patients. T 1 weighted images were obtained after intravenous administration of Gd-DTPA in all patients. Results A total of 7 ISCM were displayed by MR studies. The tumor occurred in thoracic cord (3/7) and in medullary cone (4/7). The tumor involved the central aspect of the cord. The lung cancer ISCM showed hypointensity (n=1) or isointensity (n=6) on T 1WI and hyperintensity on T 2WI. The extensive cord enlargement with cyst formation or syringomyelia was common. On contrast study, all tumors showed marked homogenous enhancement with clear borders. Conclusion The lung cancer ISCM usually presented as solitary lesions with marked contrast enhancement. The extensive cord enlargement was common in all patients. Because of the limitation in the evaluation of lung cancer ISCM on MRI, definite diagnosis of lung cancer ISCM depended on combination of clinical and MRI data.
7.Changes of bone mineral density in association with serum interleukin-6 in adolescent idiopathic scoliosis
Jie WU ; Yong QIU ; Le ZHANG ; Yanfang SUN ; Xin CHEN
Chinese Journal of Tissue Engineering Research 2005;9(10):223-225
BACKGROUND: There has been increasing attention in the study of adolescent idiopathic scoliosis (ALS)and bone metabolism, which is accompanied by osteopenia and osteoporosis. Interleukin(IL) -6, a cytokine that strongly promotes bone resorption, participates in the regulation of bone metablism.OBJECTIVE: To explore the relationship between changes of bone mineral density(BMD) and serum IL-6 content in AIS.DESIGN: A controlled non-randomized concurrent study involving patients with AIS and and healthy volunteers.SETTING: The clinic and wards of the department of spinal surgery of a university-affiliated hospital.PARTICIPANTS: Thirty-six patients with AIS(6 males and 30 females aged 12 to 18 years) were treated in the Department of Spinal Surgery, Drum Tower Hospital, Medical College of Nanjing University from July to October 2003, who had a Cobb angle ranging from 34° - 109° and curvature of the thoracic spine. Thirty-six healthy adolescent volunteers(7 males and 29 females within the range of 13 - 18 years old) served as the control group.METHODS: The BMD was measured at L2- L4 and the proximal femur;(including the femoral neck, greater trochanter and Ward' s triangle) by dual-energy X-ray absorptiometry and serum IL-6 determined by enzyme-linked immunosorbent assay(ELISA).MAIN OUTCOME MEASURES: ① BMD of the lumbar spine and femur ② Comparison of serum IL-6 content of all the subjects.RESULTS: The BMD of AIS patients at L2- L4, femoral neck, greater trochanter and Ward' s triangle was 0.79±0. 12, 0.78±0. 12, 0.65± 0. 10, and 0. 69 ± 0. 13 g/cm2, respectively, all significantly lower than the corresponding measurements of the control group(1.09 ± 0. 11, 0.95 ± 0. 11,0. 78 ± 0. 10, and 0. 88 ± 0. 11 g/cm2, respectively, P < 0. 001), whereas the serum IL-6 content in the patients was significantly higher than that in the control subjects ( P < 0. 005). The changes of BMD at the lumbar spine and the three sites of femur were negatively correlated with serum IL-6 in AIS patients( P < 0. 001 ), but not so in the control group( P > 0.05).CONCLUSION: The BMD is decreased and serum IL-6 elevated in patients with AIS, and excessive secretion of IL-6 might be one of the important factors of osteopenia in AIS.
8.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.
9.The inhibitory effects of rh-endostatin (YH-16) in combination with radiotherapy on lung adenocarcinoma A549 in mice and the underlying mechanisms.
Huita, WU ; Jie, DENG ; Shiying, YU ; Xin, WANG ; Yuan, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(1):108-12
In order to investigate the inhibitory effects of Endostar (rh-endostatin, YH-16) in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy, the transplantation tumor models of A549 lung adenocarcinoma were established. When the largest diameter of tumor reached 1.0 cm, all nude mice were randomly divided into 4 groups: Endostar group, radiotherapy group, radiotherapy plus Endostar (combined treatment) group, and control group (n=6 in each group). The largest diameter and the vertical diameter of tumor were measured at different time points. At the 16th day, mice were executed, and the tumors were applied to analysis of rate of tumor cell apoptosis, and the expression levels of basic fibroblast growth factor (bFGF) mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) and those of vascular endothelial growth factor (VEGF) by immunohistochemistry. The results demonstrated that the rate of tumor inhibition in combined treatment group was higher than that in other groups. And the rate of tumor cell apoptosis in combined treatment group was also higher than that in other groups. Meanwhile, the levels of bFGF mRNA and VEGF expression in combined treatment group were lower than those in other groups. It was concluded that Endostar obviously enhanced the curative effectiveness of radiotherapy on lung adenocarcinoma A549 in mice. The underlying mechanisms may involve the down-regulation of bFGF mRNA and VEGF expression to inhibit angiogenesis by Endostar and the cooperative effect of Endostar and radiotherapy to synergistically promote tumor cell apoptosis. And Endostar inhibits angiogenesis by down-regulating the expression of bFGF mRNA and VEGF.
10.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.