1.Establishment of human small cell lung cancer SH77 multidrug resistance cell line and analysis of its biological properties
Xiangdong ZHOU ; Guisheng QIAN ; Lingzhi LIU ;
Journal of Third Military Medical University 2003;0(16):-
Objective To establish a small cell lung cancer (SCLC) multidrug resistance (MDR) cell line SH77/CDDP and analyze its biological properties. Methods The SCLC MDR cell line SH77/CDDP was established by increasing gradually the dose of cisplatin, the first line chemotherapeutic drug for lung cancer. The chemosensitivities of SH77/CDDP and its parental cell line to 7 chemotherapeutic drugs were tested. Changes in cellular morphology and ultrastructure were observed. The cell growth curve and cell cycle were also determined. Results SCLC MDR cell line SH77/CDDP having different drug resistance to the 7 chemotherapeutic drugs was established successfully. Compared with that of its parental cell, the size of MDR cell was a bit bigger. The ratio of nucleus to cell plasma decreased and mitochondria, Golgi bodies, rough endoplasmic reticulums and free ribosomes increased. The finger like apophysis was obvious. The rate of cell proliferation of SH77/CDDP was similar to that of SH77, but the number of cells in S phase increased( P
2.Investigation on expression pattern of multidrug resistance-associated gene in human small cell lung cancer cell lines SH77/CDDP using gene chip technique
Xiangdong ZHOU ; Guisheng QIAN ; Lingzhi LIU
Journal of Third Military Medical University 1983;0(04):-
Objective To obtain the expression pattern of multidrug resistance-associated gene in human small cell lung cancer(SCLC)MDR cell line SH77/CDDP and provide basis for further studying the mechanism of human SCLC MDR induced by cisplatin.Methods Total RNA was isolated from SCLC MDR cell line SH77/CDDP and its parental cells,and synthesized into double-stranded cDNA,then synthesized into biotin-labeled cRNA probe by in vitro transcription.The cRNA probes were separately hybridized with Affymetrix GeneChip and human U133 set chips(U133A and U133B,containing 39 000 transcripts,including 33 000 known human genes and 6 000 unknown cDNA expression sequence tags,ESTs),and the signals were scanned by the GeneArray Scanner.The results were analyzed by bioinformatics.Results Compared with the gene expression profile of parental SH77 cells,2 389(6.13%)genes were up-regulated in SCLC MDR cell line SH77/CDDP cells,of which 461(1.19%),89(0.23%),26(0.07%)and 7(0.02%)genes were separately up-regulated one-fold,two-folds,three-folds and four-folds.36 genes,including ABCC3,HSP70,CYP26B1,CYP4A11,IGF1R,LOX2,CAP2,LRP2BP,AKNA,ELTD1 and otherwise,were up-regulated 2.5-5.1 folds.According to Gene Ontology and Tree View analysis,these 35 genes were involved in transport,cell adhesion,signal transduction,transcription and ion binding and so on.Conclusion Many multidrug resistance-associated genes induced by cisplatin have been screened by high-throughput gene chip method.Validating their cellular functions will help to identify the mechanism of human SCLC MDR induced by cisplatin.
3.Study of the drug sensitivity of primary cultured lung cancer cells and the expressions of four drug resistance related proteins
Lingzhi LIU ; Xiangdong ZHOU ; Guijun HUANG ; Guisheng QIAN ;
Journal of Third Military Medical University 2003;0(08):-
Objective To study the drug sensitivity of chemotherapeutic drugs by MTT method. Methods Twenty five lung cancer specimens obtained by operation were used for primary culture and the sensitivity of the cancer cells to four drugs, adriamycin(ADM), cis dichlorodiamine platinum(DDP), vincristine(VCR) and etoposide(VP16), was tested by MTT assay. TOPOⅡ, p53, bcl 2 and GST ? proteins in 10 cases of lung cancer tissue were determined by immunohistochemistry. Results The successfully obtained results in 21 cases showed that the effective rate of ADM, DDP, VCR and VP16 was 19.0%, 38.1%, 23.8% and 23.8% respectively. Positive expressions of TOPOⅡ, p53, bcl 2 and GST ? proteins were found in lung cancer tissue in the 10 cases which had different multi drug resistance(MDR) to the four drugs. Conclusion It is possible to test the drug sensitivity of the primary cultured lung cancer cells obtained from operation by MTT method and the results are useful for the selection of anticancer drugs. The expressions of TOPOⅡ, p53, bcl 2 and GST ? are factors to determine MDR for lung cancer cells.
4.Risk factors of the thrombopenia in septic patients complicated with acute kidney injury
Lingzhi JIANG ; Mingshan WANG ; Jinzhu WANG ; Renhua SUN ; Qian LI
Chinese Journal of Clinical Infectious Diseases 2017;10(3):187-193
Objective To investigate the risk factors of thrombopenia(TP)in septic patients complicated with acute kidney injury (AKI).Methods Two hundred and sixty five septic patients complicated with AKI admitted in Intensive Care Unit ICU of Zhejiang Provincial People''s Hospital during January 2012 and December 2016 were enrolled in the study.The clinical data, results of laboratory tests, Acute Physiology and Chronic Health Evaluation (APACHEII) scores, Sequential Organ Failure Assessment (SOFA) scores, therapeutic intervention, and 28-day mortality were documented.Among 265 patients, TP occurred within 7 days in 112 cases (TP group) and did not occur in 153 cases (non-TP group).Multivariable Logistic regression analysis was performed to analyze the risk factors of TP.Results The 28-day mortality rate in TP group was higher in TP group than that in non-TP group (47.3% vs.33.3%, χ2=5.307,P<0.05).Univariate analysis showed that age, C-reactive protein (CRP), procalcitonin (PCT) and APACHEII score, SOFA score, continuous renal replacement therapy (CRRT), heparin anticoagulation, shock, usage of linezolid and bloodstream infections were associated with TP in septic patients with AKI(all P<0.05).Multivariable Logistic regression analysis showed that age≥65 (OR=4.53, 95%CI 1.23-9.24,P<0.05), CRRT(OR=5.24,95%CI 2.14-14.56,P<0.01), heparin anticoagulation(OR=4.56,95%CI 2.13-8.46,P<0.01), usage of linezolid(OR=2.35,95%CI 1.25-5.24,P<0.01), shock(OR=2.15,95%CI 1.03-4.96,P<0.01)and bloodstream infections(OR=4.26,95%CI 1.36-12.48,P<0.01)were independent risk factors for septic patients with TP.Conclusion For septic patients with AKI having these risk factors, the platelet counts should be closely monitored, and intervention measures should be given to reduce the occurrence of TP.
5.Effect of Ultrasound-guided Subcostal Transverses Abdominis Plane Block with Dexmedetomidine after Laparoscopic Radical Operation on Colon
Yuan ZHOU ; Jiashu QIAN ; Lingzhi XUE ; Su CAO ; Jingjing DING
Chinese Journal of Rehabilitation Theory and Practice 2014;20(12):1171-1174
Objective To investigate the effect of ultrasound-guided subcostal transverses abdominis plane (TAP) block with dexmedetomidine after laparoscopic radical operation. Methods 40 patients underwent laparoscopic radical operation for colorectal cancer were randomized into dexmedetomidine group (group DEX) and control group (group CON). All the patients received ultrasound-guided subcostal TAP block after operation, Group DEX with dexmedetomidine 1 μg/kg and 0.25% ropivacaine to 20 ml, and group CON with 0.25% ropivacaine 20 ml. All the patients were assessed with Ramsay scores and the pain at rest and on coughing were assessed with Visual Analogue Scale (VAS), 2, 6, 12, 24 and 24 hours after operation. The highest level and the duration of sensory blockade, the first time and the total times of pressing the analgesia pump in the first day after operation, and the requirements of sufentanil were recorded. First flatus time, first diet time and the length of hospital stay were compared. Results The scores of VAS were significantly less (P<0.001), and the Ramsay scores were more in the group DEX than in the group CON (P<0.01) 2, 6 and 12 hours after operation; with the longer time of sensory blockade (P<0.001), the later to first press the analgesia pump (P<0.001), the less frequence of pressing the analgesia pump (P<0.001), and less dosage of sufentanil (P<0.001). The first flatus time, first diet time were significantly earlier in the group DEX than in the group CON (P<0.001), with the less length of total hospital stay (P<0.001). Conclusion Dexmedetomidine can promote the anaesthesia of ultrasound-guided subcostal TAP block with ropivacaine and improve the recovery after laparoscopic radical operation.
6.Analysis of fusion gene expression in acute myeloid leukemia
Qi YAN ; Yani LIN ; Xianqi HUANG ; Lingzhi QIAN ; Jingting MA ; Hong ZHANG ; Long CHEN ; Xuejing CHEN ; Yingchang MI ; Kun RU
Chinese Journal of Hematology 2021;42(6):480-486
Objective:To analyze the genetic landscape of multiple fusion genes in patients with de novo acute myeloid leukemia (AML) and investigate the characteristics of immunophenotypes and mutations.Methods:The results of multiple fusion genes from 4192 patients with de novo AML were retrospectively analyzed from 2016 to 2020. In addition, the immunophenotypical data and the mutational results from high-through put method were statistically investigated and correlated as well.Results:①Among the 52 targets, 29 different types of fusion genes were detected in 1948 patients (46.47%) with AML, which demonstrated an "exponential distribution" . ② As the age increased, the number of patients with fusion gene increased first and then decreased gradually. The total incidence rate of fusion genes and MLL rearrangment in children were significantly higher than those in adults (69.18% vs 44.76%, 15.35% vs 8.36%) . ③The mutations involving FLT3 and RAS signaling pathway contributed most in patients with MLL rearrangment. ④No specific immunophenotypic characteristics were found in AML patients with MLL or NUP98 rearrangements. Conclusion:Nearly half of AML patients were accompanied by specific fusion gene expression, the proportions of different fusion genes in pediatric and adults patients were different by multiple PCR. The gene mutations and immunophenotype of these AML patients have certain rules.