1.Activated Notch1 signaling inhibits growth of EC109 cell line and its mechanism
Yongli ZHANG ; Kejie ZHANG ; Xianghui MIN ; Quanyi LU ; Wenli LIU
China Oncology 2009;19(8):597-601
Background and purpose: It has been reported that activation of Notch1 could strongly inhibit proliferation of HPV (human papilloma virus)-positive HeLa cells by down-regulation of the E6 and E7 genes. The aim of this paper was to investigate the role of the Notch signaling pathway in growth arrest of EC109 cells in vitro and the molecular mechanism. Methods: EC109 cell lines, a well differentiated human ESCC (esophageal squamous cell carcinoma) cell line with HPV18-positive, was used in the study. Exogenous intracellular domain of Notch1(ICN) was transfected into cultured EC109 cells by lipofectamine transfection, the proliferation of the transfected cells was measured by an MTT assay. Cell cycle distribution was analyzed by flow cytometry. Human papilloma virus type 18 (HPV18) E6/E7 mRNA expression was detected by RT-PCR, and p53 protein expression was detected by Western blot.Results: Activation of Notchl signaling resulted in inhibition of EC109 cell proliferation with the induction of G_2/ M arrest. There was a significant difference in terms of the percentage of G_2/M phase cells among the ICN-transfected group (42.57±1.57)% and the non-transfected group (1.88±0.66)% or the empty plasmid transfected group (1.99±1.02)% (P<0.01). Down modulation of HPV18 E6/E7 gene expression and upregulation of p53 expression was (2.15±0.23) in ICN-transfected group higher than non- transfected group (0.45±0.07) and empty plasmid transfected group (0.46±0.02) (P<0.01). Conclusion: Repression of HPV18 E6/E7 expression by Notch1 signaling results in growth suppression of HPV18-positive EC109 cells with concomitant activation of p53-mediated pathways.
2.Changes of cerebra flow under monitoring of transcranial Dopple in posterior retroperitoneoscopic surgery
Jixiu XUE ; Yang HUA ; Kejie WANG ; Weitao CHENG ; Qinghai LIU ; Limin WEI ; Jingsheng LI
Journal of Medical Postgraduates 2003;0(07):-
Objective:To explore the changes of cerebra flow in retroperitoneoscopic surgery.Methods: Twenty-two patients undergoing retroperitoneoscopic surgery were selected.We measured Vm、Delt and PI with transcranial Dopple(TCD),and measured endothelin(ET),neurol peptide Y(NPY),Calcitonin gene-related peptide(CGRP) in plasma before CO2 insufflation,after CO2 insufflation,and after CO2 deflation.Results:Vm and Delt increased significantly at 30 min after pneumoperitonenum,and reached the peak at 90 min after pneumoperitonenum.PI decreased significantly 90 min after pneumoperitonenum.The plasma concentrations of ET and NPY increased significantly 60 min after pneumoperitonenum. Conclusion:Cerebra flow changes significantly in patients undergoing retroperitoneoscopic surgery,which is correlated to the changes of plasma CO2 and vascular bioactive substances.
3.An Analysis of the Common Deafness Gene Mutation from 356 Non-Syndromic Deafness Patients in Guizhou Province
Xue YANG ; Youqin WANG ; Hongyuan GUO ; Li LAN ; Yuqing LIU ; Zuwei CAO ; Fangying TAO ; Kejie YANG
Journal of Audiology and Speech Pathology 2017;25(1):9-13
Objective The study was to investigate the spectrum and incidence of the hot-spot deafness gene mutations from non-syndromic hearing loss(NSHL)in Guizhou province.Methods Genomic DNAs of 356 patients of NSHI in Guizhou were obtained from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the gene GJB2,SLC26A4,GJB3 and mitochondria 12SrRNA.Results Eighty-eight (24.72%)out of 356 patients were found carrying at least one pathogenic gene mutation.Among them,40 patients with the GJB2 mutations were found(11.24%),including 19 cases(5.34%)of homozygous matutions,5 cases(1.40%)of com-pound heterozygous mutations,and 15 cases(4.21%)of single heterozygous mutations.Twenty-nine patients with the SLC26A4 mutations were found (8.15%),including 9 cases(2.53%)of homozygous mutations and 19 cases (5.34%)of single heterozygous mutations.Homogenic mitochondrial 12SrRNA 1555A>G mutations were in 10 patients(2.81%)and 1494C>T mutations in 10 patients (0.56%),and heterogeneous mitochondrial 12SrRNA 1555A>G mutations in 7 patients(1.97%).GJB3 gene c.538C>T heterozygous mutations was found in 1 patients. Conclusion The mutations of GJB2 and SLC26A4 gene are two major pathogenic genes for patients with NSHL in Guizhou province.Our study provides a theoretical basis for the early diagnosis,genetic counseling and treatment of deafness.
4.Test of urine leukotriene FA in infants with bronchiolitis and its clinical value
Huanyin YAO ; Shumei LIU ; Guozheng ZHU ; Xiaohong CHEN ; Kejie XIE ; Wenyong LOU ; Wei WANG ; Xiaoxian WANG
Journal of Clinical Pediatrics 2010;(2):152-155
Objective To study the prognosis of infants with bronchiolitis by testing urine leukotriene E4 (LTE4) level and investigating atopy's influences. Methods Urine LTE4 was tested in 38 eases with mild bronchiolitis (47 in acute stage, 17 in convalescent stage), 9 severe bronchiolitis cases, 15 atopic cases, 25 control cases. Peripheral blood was used to determine eosinophils count (EC) in acute bronchiolitis cases. Results (1) The level of urine LTE4 is obviously higher in cases of acute group (62.11 ± 12.23 pmol/L) than that of control group (22.19±1.50 pmol/L) , and the convalescent group (34.86 ±5.75 pmol/L) (F = 132.42, P < 0.01) ;Urine LTE4 level of convalescent group is higher than that of the control group (P < 0.01). (2) Urine LTE4 level is significantly higher in severe group (98.04 ± 8.04 pmol/L) than that of mild group (59.16 ± 12.25 pmol/L) (t = 9.92, P < 0.01). (3) Urine LTE4 level of atopy positive (88.75 ± 10.45 pmol/L) infants with bronchiolitis is significantly higher than atopy negative infants (55.28 ± 11.44 pmol/L)(t = 8.63, P < 0.01). (4) There is no significant correlation between the levels of urine LTE4 and EC for acute bronchiolitis. Condusions The level of urine LTE4 in acute bronehiolitis patients increases and remains high in convalescent stage;Higher urine LTE4 level in severe bronchiolitis cases indicates that urine LTE4 level is related to the severity of the disease;cysteinylleukotrenes is an important mediator of inflammation that may influence the prognosis of atopy positive infants with bronchiolitis;EC is not a good index to present the airway inflammation of infants with bronehiolitis.
5.Clinical analysis of 68 cases of small intestine bleeding
Kejie LIU ; Shilun TONG ; Yongbin ZHENG ; Hongfa GAN ; Fengyu CAO ; Xiaobo HE ; Yu DING
Clinical Medicine of China 2012;28(3):307-309
Objective To investigate the causes,diagnosis and treatment of small intestine bleeding.Methods Sixty-eight cases of small intestine bleeding from January 2000 to June 2010 were retrospectively analyzed.Among all cases,4 underwent routine hemostatic treatment under colonoscopy,40treated with open surgery and 24 patients with laparoscopic therapy.Among them,57 cases underwent part resection for some small intestine,completely laparoscopic resection of diverticula was performed in 7patients.Results Neoplasms was the leading cause of small intestine bleeding,accounting for 48.5% (33/68)in these patients,followed by small intestine diverticulum accounted for 29.4% ( 20/68 ),intestinal infective diseases accounted for 14.7% ( 10/68 ) and vascular disease accounted for 7.4% ( 5/68 ).Conclusion The clinical manifestations of small intestinal bleeding showed no specific signs.Neoplasm,intestine diverticulum and intestinal infective diseases are the most common causes of small intestinal bleeding.Small intestinal bleeding can be diagnosed in intraoperative colonoscopy.Surgery is the most effective treatment for small intestinal bleeding.
7.Irradiation injury temporarily induces enhancement of APN/CD13 peptidase activity on aorta-gonads-mesonephros (AGM)-derived stromal cells.
Yan, ZHU ; Lifang, HUANG ; Xiaohua, LUO ; Hanying, SUN ; Dan, RAN ; Kejie, ZHANG ; Miao, ZHENG ; Kun, ZHOU ; Wenli, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):145-8
This study was designed to investigate the expression of aminopeptidase N (APN)/CD13 on intraembryonic AGM stromal cells, and the change of its enzymatic activity after irradiation injury. The expression of APN/CD13 on AGM stromal cells was assayed by RT-PCR and immunihistochemistry. After the stromal cells in AGM region were irradiated with 8.0 Gy of (60)Co gamma-rays, APN/CD13 enzymatic activity was measured by spectrophotometer at different time points. The result showed that AGM stromal cells strongly expressed APN/CD13. The enzymatic activity of APN/CD13 decreased temporarily after irradiation injury, then increased to higher level 4 h after irradiation, and it returned to the pre-irradiation level 24 to 48 h after the irradiation. The enzymatic activity of APN/CD13 was temporarily enhanced after irradiation injury, which might be one of the compensatory mechanisms that promote the hematopoietic recovery after irradiation.
8.Comparison of Rehabilitation Effects of Deaf Children with Different Compensation Strategies
Li LAN ; Fei LI ; Yuqing LIU ; Qing YE ; Kejie YANG ; Min WANG ; Fangying TAO ; Wei HAN ; Yuan ZHANG ; Youqin WANG
Journal of Audiology and Speech Pathology 2015;(2):186-190
Objective The hearing and speech rehabilitation effects by hearing aid use and cochlear implanta‐tion in hearing -impaired children were compared .Methods A total of infants aged from 6 months to 3 years with severe hearing loss of prelingual nature were included .They were divided into 2 groups :group 1 (18 with hearing aids) and group 2 (14 with cochlear implantation) .There was no significant difference in the degree of hearing loss and age between the 2 groups .They received rehabilitation education of the same content for a year .Their unaided and aided thresholds were tested in sound field while the IT -MAIS was used for evaluating hearing ability of two groups before and after 3 ,6 ,9 ,and 12 months of hearing aid use and cochlear implantation .Results In both groups ,hearing were better in aided conditions than those of in unaided ,while those from the cochlear group were significantly improved at more frequencies than those of the hearing aid group .In general ,the longer use and train‐ing ,the better hearing improvement .The mean scores of auditory performance in the cochlear implantation group at 6 months were significantly higher in comparison with those at 3 months (P<0 .05) .There were significant differ‐ences in mean scores between the two groups at 6 ,9 ,12 months after fitting (P<0 .05) .The verbal ability scores ,hearing and speech rehabilitation levels in the cochlear group were higher than those of in the hearing aid group (P<0 .05) .Conclusion For severe hearing loss children ,cochlear implantation could produce more noticeable effects than hearing aids .
9.Enhancing 2-keto-L-gulonic acid production under hyperosmotic stress by adding sucrose.
Kejie CHEN ; Jingwen ZHOU ; Liming LIU ; Jie LIU ; Guocheng DU ; Jian CHEN
Chinese Journal of Biotechnology 2010;26(11):1507-1513
This study aimed to further enhance 2-keto-L-gulonic acid (2-KLG) production efficiency. A strategy for enhancing Ketogulonigenium vulgare growth and 2-KLG production by improving B. megaterium growth with sucrose was developed based on the time course of osmolality during 2-KLG industrial scale fermentation and effects of osmolality on cells growth and 2-KLG production. Results showed that the accumulation of 2-KLG and the feeding of alkaline matter led to an osmolality rise of 832 mOsmol/kg in the culture broth. High osmotic stress (1 250 mOsmol/kg) made the growth ofB. megaterium and K. vulgare decreased 15.4% and 31.7%, respectively, and consequently the titer and productivity of 2-KLG reduced 67.5% and 69.3%, respectively. When supplement sucrose under high osmotic condition (1 250 mOsmol/kg), B. megaterium growth was significantly improved, with the result that 2-KLG production was increased 87%. Furthermore, by applying this sucrose addition strategy further to batch fermentation in 3 L fermentor, the productivity of 2-KLG increased 10.4%, and the duration of fermentation declined 10.8%. The results presented here provide a potential strategy for enhancing the target metabolites produced by mixed strains at environmental stress.
Bacillus megaterium
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genetics
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growth & development
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metabolism
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Fermentation
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Industrial Microbiology
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Osmosis
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Rhodobacteraceae
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genetics
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growth & development
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metabolism
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Stress, Physiological
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Sucrose
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pharmacology
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Sugar Acids
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metabolism
10.Enhancement of 2-keto-L-gulonic acid production using three-stage pH control strategy.
Jing ZHANG ; Jingwen ZHOU ; Liming LIU ; Jie LIU ; Kejie CHEN ; Guocheng DU ; Jian CHEN
Chinese Journal of Biotechnology 2010;26(9):1263-1268
The aim of this study was to improve the 2-keto-L-gulonic acid (2-KLG) production efficiency by Ketogulonicigenium vulgare and Bacillus megaterium by using multi-stage pH control strategy. The effect of pH on the cell growths and 2-KLG production showed that the optimum pH for K. vulgare and B. megaterium cell growth were 6.0 and 8.0, respectively, while the optimum pH for 2-KLG production was 7.0. Based on the above results, we developed a three-stage pH control strategy: the pH was kept at 8.0 during the first 8 h, then decreased to 6.0 for the following 12 h, and maintained at 7.0 to the end of fermentation. With this strategy, the titer, productivity of 2-KLG and L-sorbose consumption rate were achieved at 77.3 g/L, 1.38 g/(L x h) and 1.42 g/(L x h), respectively, which were 9.7%, 33.2% and 25.7% higher than the corresponding values of the single pH (pH 7.0) control model.
Bacillus megaterium
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growth & development
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metabolism
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Culture Media
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chemistry
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Fermentation
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Hydrogen-Ion Concentration
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Rhodobacteraceae
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growth & development
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metabolism
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Sorbose
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metabolism
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Sugar Acids
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metabolism