1.Detection of truncating mutations of APC gene by digital protein truncation test labeled with fluorescent Lys-t-RNA
Xin ZHANG ; Hai LI ; Yong DU ; Xing GAO ; Yinxue YANG
Chinese Journal of Digestive Surgery 2012;11(4):386-390
Objective To investigate the relationship between truncating mutations of APC gene and sporadic colorectal cancer,and analyze the feasibility of non-radioactive protein truncation test (PTT) in the detection of mutations of APC gene.Methods Ninety-six samples of sporadic colorectal cancer tissues ( including 44 patients with colonic cancer and 52 patients with rectal cancer) were obtained from the General Hospital of Ningxia Medical University from September 2008 to September 2010.The mutation cluster region of the APC gene was screened using digital PTT labeled with fluorescent Lys-t-RNA,with a polymerase chain reaction fragment amplified from genomic DNA serving as a tenplate for in vitro translation.The occurrence of gene mutation was determined according to the emergence of truncated peptides.The mutation cluster region of the APC gene in 46 samples of colorectal cancer tissues was analyzed by direct sequencing.The detection rates of the 2 methods were compared by chi-square test.Results Thirteen (26%) truncated peptides were detected in the 50 samples of colorectal cancer tissues.The mutation type of 4 samples is nonsense mutation,which resulted in emergence of truncated gene products.Eleven (24%) truncated peptides were detected in the 46 samples of colorectal cancer tissues.There was no significant difference in the detection rates between PTT and direct sequencing ( x2 =0.033,P > 0.05 ).Conclusions Truncating mutations of APC gene are common alterations in sporadic colorectal cancer in the Chinese.Digital PTT labeled with fluorescent Lys-t-RNA is rapid and high-sensitive in screening gene mutations.
2.Expression of Truncated NSP2 Protein of Porcine Reproduction and Respiratory Syndrome Virus in E.coli and Preparation of Monoclonal Antibodies Against NSP2 Protein
Hai-Yan WANG ; Ping JIANG ; Yi-Jun DU ; Yu-Feng LI ; Jun-Xing LI ; Fang SHEN ;
China Biotechnology 2006;0(02):-
The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.
3.Psychologic status and their influencing factors in congenital microtia patients and their families.
Jia-mei DU ; Hong-xing ZHUANG ; Hai-yue JIANG ; Bo PAN ; Wan-hou GUO ; Xue-chuan LI
Chinese Journal of Plastic Surgery 2005;21(3):218-221
OBJECTIVETo study the psychologic status and their influencing factors in congenital microtia patients and their families.
METHODSTotally one hundred and two congenital microtia patients (79 men, 23 women, mean age 13.62 +/- 7.2 years) were enrolled. The patients and their families answered the questionnaire written by ourselves to identify the psychosocial problems.
RESULTS(1) 23.5% patients were found to have severe psychosocial problems, such as lack of self-confidence, close and fear and so on. (2) With the growth of age, psychosocial problems of the patients were rated high (P < 0.05). (3) For patients who found their deformations early, psychosocial problems also were rated low. (4) For patients who found their deformations by themselves, psychosocial problems also were rated low. (5) The education and psychosocial impact for parents all affected patients deeply.
CONCLUSIONSTo prevent psychosocial problems, we should operate for patients as early as possible. And correct guidance is very important for youngsters.
Adolescent ; Adult ; Child ; Child, Preschool ; Congenital Abnormalities ; epidemiology ; psychology ; Ear ; abnormalities ; Family ; psychology ; Female ; Humans ; Inpatients ; statistics & numerical data ; Male ; Surveys and Questionnaires ; Young Adult
4.Effect of sphingosine kinase 1 on the apoptosis, migration and invasion of colon cancer HT-29 cells and its molecular mechanisms.
Shi-quan LIU ; Meng-bin QIN ; Jie-an HUANG ; Yue-yuan ZHONG ; Guo-du TANG ; Hai-xing JIANG
Chinese Journal of Oncology 2011;33(3):178-182
OBJECTIVETo investigate the effect of sphingosine kinase 1 (SphK1) on the proliferation, apoptosis, migration and invasion of colon cancer TH-29 cells and to explore its molecular mechanisms.
METHODSPhorbol 12-myristate 13-acetate (PMA) was used to induce the activity of SphK1 and N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. Cell prolieration and apoptosis were detected by MTT assay and flow cytometry, respectively. The migration and invasion capabilities of the cells were assessed in Transwell chambers. The activity of SphK1 was assayed by autoradiography. Western blot was used to evaluate the protein expression of SphK1, p38, phosphorylated p38 (p-p38) and SAPK/JNK.
RESULTSPMA and DMS were able to induce and suppress the activity and protein expression of SphK1 in a time-dependent manner, respectively. PMA enhanced and DMS suppressed the cell viability in a time- and dose-dependent manner. Being treated with 100 nmol/L PMA or 50 µmol/L DMS for 0, 6, 12, 24 h, the cell apoptosis rates of PMA group were (9.35 ± 0.84)%, (7.61 ± 0.48)%, (5.53 ± 0.76)% and (0.56 ± 0.33)%, contrastly, that of DMS group were (9.18 ± 0.94)%, (12.06 ± 1.41)%, (19.80 ± 2.36)% and (31.85 ± 3.60)%, respectively. Compared with the control group, the cell migration and invasion capabilities of the PMA group were significantly enhanced, and that of the DMS group were significantly suppressed. The migration cell number of control, PMA and DMS groups were 68.75 ± 6.15, 109.33 ± 11.63 and 10.83 ± 2.48, the invasion cell number of control, PMA and DMS groups were 55.42 ± 4.50, 90.58 ± 7.06 and 9.58 ± 2.39, respectively. With the elevating activity and expression of SphK1, the protein expressions of p38, p-p38 and SAPK/JNK were strikingly suppressed. On the contrary, after treating with DMS the protein expressions of p38, p-p38 and SAPK/JNK were enhanced.
CONCLUSIONSSphK1 potently enhances the prolieration, migration and invasion of colon cancer HT-29 cells, meanwhile suppresses the cell apoptosis. The suppressing of the p38 and SAPK/JNK signalling pathways may be one of its molecular mechanisms.
Apoptosis ; drug effects ; Carcinogens ; administration & dosage ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; HT29 Cells ; Humans ; MAP Kinase Kinase 4 ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; physiology ; Sphingosine ; administration & dosage ; analogs & derivatives ; pharmacology ; Tetradecanoylphorbol Acetate ; administration & dosage ; analogs & derivatives ; pharmacology ; Time Factors ; p38 Mitogen-Activated Protein Kinases ; metabolism
5.Effects of Long-term Treatment with Hydrochlorothiazide Combined Spironolactone or Captopril on Left Ventricular Hypertrophy in Hypertensive Patients
Ai-Jun XING ; Dong-Xian LI ; Xin DU ; Shou-Ling WU ; Hai-Yan ZHAO ; Li-Ming LING ; Dong-Qing LI ; Zheng-Xin CAO ; Gui-Ling WANG ; Qing YU ;
Chinese Journal of Hypertension 2007;0(05):-
0.05);2)After 12,24,36 months' treatment,BP was decreased significantly in each group (P0.05).Conclusion Both combined spirono- lactone/HCTZ and captopril/HCTZ significantly reduced BP and LVMI or LVMI and the maguitude of reduction was further enhanced after prolonged treatment.
6.Complete genome sequencing of enterovirus 71 (EV71) HENAN08 strain isolated in Henan province
Shao-Li ZHANG ; Bian-Li XU ; Wan-Shen GUO ; Li CHEN ; Hai-Yan WEI ; Yan-Hua DU ; Xing-Le LI
Chinese Journal of Epidemiology 2009;30(9):938-941
strains (isolated in 2008). Conclusion The HENAN08 strain might belong to the same genogroup with AnhuiFY08 and Zhejiang08 strains as C4 gene subtypes.
7.Identification and Characterization of Gonatobotryum apiculatum CausingLeaf Spot and Blight on Sinowilsonia henryi
Ying GAO ; Hai Feng LIU ; Zheng Xing SONG ; Xiao Ying DU ; Jian Xin DENG
Mycobiology 2020;48(1):70-74
Sinowilsonia henryi is a rare and endangered plant, as well as an endemic species in China.In July 2018, leaf spot and blight disease was observed on S. henryi in Yichang, Hubei,China. A fungus isolated from disease tissues was identified as Gonatobotryum apiculatumbased on morphology and sequence analyses of ITS and LSU regions. Phylogenetic analysesindicated that the species belongs to Dothioraceae (Dothideales). Morphologically, the speciesproduced two distinct types of conidia from authentic media, both conidia weredescribed here. Pathogenicity tests showed that the fungus is a pathogen causing leaf spotson S. henryi. This is the first report of leaf spot and blight disease on S. henryi caused byG. apiculatum in China.
8.Identification and Characterization of Gonatobotryum apiculatum CausingLeaf Spot and Blight on Sinowilsonia henryi
Ying GAO ; Hai Feng LIU ; Zheng Xing SONG ; Xiao Ying DU ; Jian Xin DENG
Mycobiology 2020;48(1):70-74
Sinowilsonia henryi is a rare and endangered plant, as well as an endemic species in China.In July 2018, leaf spot and blight disease was observed on S. henryi in Yichang, Hubei,China. A fungus isolated from disease tissues was identified as Gonatobotryum apiculatumbased on morphology and sequence analyses of ITS and LSU regions. Phylogenetic analysesindicated that the species belongs to Dothioraceae (Dothideales). Morphologically, the speciesproduced two distinct types of conidia from authentic media, both conidia weredescribed here. Pathogenicity tests showed that the fungus is a pathogen causing leaf spotson S. henryi. This is the first report of leaf spot and blight disease on S. henryi caused byG. apiculatum in China.
9.The observation on effectiveness of different administration time and positions of Kaiselu for patients with acute myocardial infarction on prevention of their constipation
Hai-Bo GU ; Mei-Fang DU ; Yue-Zhen ZOU ; Lin CHEN ; Xing-Zhu WEN ; Li-Yao ZHUANG
Chinese Journal of Modern Nursing 2008;14(11):1304-1305
Objective To observe the effectiveness of different administration time and positions of Kaiselu for patients with acute myocardial infarction On prevention of their constipation.Methods 68 acute myocardial infarction patients without three days puzzles stool were randomly divided into two groups f 34 case8 in each group).The patients in the observation group received Kaiselu 40~50 ml without the awareness of defecation,by the anal insertion 7~10 cm,while the control group received Kaiselu 40~50 ml with the awareness of defecation, by the anal insertion 3~4 cm. Results The defecation effect in the observation group Wag significantly higher than the control group(P<0.01).Conclusions It shows better defecation effect for patients with acute myocardial infarction received Kaiselu without the awareness of defecation by the anal insertion 7~10 cm.