1.Effects of basic fibroblast growth factor on expressions of collagen and fibronectin in normal skin and hypertrophic scar fibroblasts
Rui SONG ; Huining BIAN ; Wen LAI ; Dehua CHEN ; Kesen ZHAO
Chinese Journal of Tissue Engineering Research 2010;14(41):7784-7790
BACKGROUND:Basic fibroblast growth factor(bFGF)can promote production of collagen,fibronectin and matrix enzyme in healing wounds.However,dysregulation of this process,such as the abnormal coordination of cell proliferation,extracellular.matrix and neovasculadzation formation,or remodeling of the wound matrix will lead to excess accumulation of scar tissues.OBJECTIVE:To investigate effects of bFGF on normal skin wound healing and hypertrophic scar formation.METHODS:Normal and hypertrophic scar fibroblasts from tissue biopsies from 5 patients who underwent plastic surgery for repairing hypertrophic scars were isolated and cultured.The expressions of collagen,fibronectin and protein synthesis were detected by RT-PCR and ELISA.The mitochonddal membrane potential changes were measured using JC-1 staining and flow cytometry.Simultaneously,adenosine tdphosphate(ATP)levels were determined by chemiluminescence method.The effects of bFGF on these indexes of normal and hypertrophic scar fibroblasts were observed.RESULTS AND CONCLUSION:Hypertrophic scar fibroblasts become slower after being exposed to bFGF,which selectively inhibited type Ⅰ collagen production in hypertrophic scar fibroblasts(P<0.05).Although bFGF inhibited type]collagen production,it had no effect on type Ⅲ collagen expression in both normal and hypertrophic scar fibroblasts.However,fibronectin expression in the normal fibroblasts was up-reguleted after bFGF treatment(P<0.05).In addition,the mitochonddal membrane potential tended to depolarization,although no statistical difference,in hypertrophic scar fibroblasts treated with bFGF(10 or 100 μg/L).bFGF treatment increased the cellular ATP levels in the normal fibroblasts,while there were no significant alterations in the hypertrophic scar fibroblasts over a treatment of bFGF(10 or 100 μg/L,P<0.05).The results suggest that there are differential effects and mechanisms on the skin fibroblasts with bFGF treatment in normal wound healing and hypertrophic scar formation.
2.Karyotype analysis of chorionic villi from pregnant women with missed abortion using multiplex ligation-dependent probe amplification
Yan GU ; Jiansheng XIE ; Fuwei LUO ; Qian GENG ; Huakun ZHANG ; Huining SHEN ; Kun ZHAO ; Qingzhi LIU
Chinese Journal of Obstetrics and Gynecology 2009;44(7):509-513
Objective To evaluate the clinical value of multiplex ligation-dependent probe amplification (MLPA) technique used in karyotype analysis of chorionic villi from missed abortion. Methods Feb 2008 to Oct 2008, 91 patients with missed abortion diagnosed by hormonal measurement, type B ultrasound and physical exam matched with 20 normal pregnant women undergoing artificial abortion were enrolled in this study. Chorionic villi was obtained by suction dilation and curettage in aseptic condition, then those villi was cultured and analyzed by traditional cytogenetic karyotyping method, in the mean time, the DNA extracted from villi was detected by MLPA. The results of chromosomal G-banding of chorionic villi were compared between two methods. Results The diagnostic concordance of MLPA and traditional karyotyping was observed in 92% (84/91) cases, there were 84 cases in the case group with diagnostic concordance by traditional karyotyping and MLPA except 7 cases of euploidy could not be detected by MLPA. The 84 cases included 40 normal karyotype,29 trisomy of euchromosome, 1 double trisomy of euchromosome, 10 monosomy X , 1 monosomy X combined with trisomy of euchromosome, 2 chimaera of X chromosome, 1 structural abnormity of euchromosome. Among 7 cases with discordance diagnosis, 2 cases with trisomy and 5 cases with tetrasomy of euchromosome were identified in traditional karyotyping, however, they were all diagnosed with normal disomy by MLPA. Of 20 villi from normal pregnancy, two methods got the consistent results. Conclusion The MLPA was rapid and efficacy method used for analyzing aneuploids in chorionic villi.
3.Study on sleeping quality of medical students and its influential factors
Xiao ZHENG ; Chichen ZHANG ; Huining ZHAO ; Xiaojie PAN ; Weihong HU ; Chunhui SU
Chinese Journal of Health Management 2017;11(3):240-244
Objective To understand the quality of sleeping in medical students, and provide a reference basis for designing sleep health management program for college students. Methods The survey was conducted at a medical college in Shanxi province with stratified random sampling and Pittsburgh Sleep Quality Index (PSQI) scale. Totally 210 individuals were enrolled, 199 completed the questionnaire, with a response rate of 93.5%. SPSS 21.0 statistical software was used for data analysis. Descriptive statistical analysis was carried out for socio-demographic data. Independent t-test, one-way analysis of variance and non-parametric test were used to compare the scores of PSQI between participants. Multiple stepwise linear regression was used to determine influencing factors of sleep quality. Results The average score of PSQI for the 199 medical students interviewed was (6.74±2.42), the incidence of sleep disorder was 50.25%, the overall sleep quality was not high. Learning pressure of medical student had statistically significant in PSQI (Completely unmatched, 5.14 ± 2.32, Mostly unmatched, 5.93 ± 2.73, Mostly matched, 6.70 ± 2.42, Absolutely matched, 7.39 ± 2.10, F=3.366, P=0.020), employment pressure of medical students in PSQI (Completely unmatched, 6.50 ± 0.71, Mostly unmatched, 6.80 ± 3.24, Mostly matched, 5.98 ± 2.14, Absolutely matched, 7.14 ± 2.35, F=3.134, P=0.027). Grade (β=-0.172, P<0.05) and learning pressure (β=0.210, P<0.01) were the main influence factors for students. Conclusion Sleep quality of medical students was not high, upper grade was worse than low grade, the incidence of sleep disorder had increasing trend as learning pressure and degree of employment pressure worsen.
4.Analysis of the prevalence and related factors for comorbidity of myopia, obesity, and depression symptoms among middle school students in Beijing
LUO Huijuan, SUN Bingjie, ZHAO Hai, CHEN Dongni, WANG Lu, GAO Ruoyi, KUANG Huining, E Boran, GUO Xin
Chinese Journal of School Health 2024;45(7):1045-1049
Objective:
To analyze the current status and related factors of comorbidity of myopia, obesity, and depression symptoms among middle school students in Beijing, so as to provide a basis for comprehensive public health interventions for common diseases.
Methods:
Through stratified cluster random sampling in October 2022, a total of 11 262 junior high school, senior high school, and vocational high school students in 16 districts of Beijing were surveyed with self administered questionnaires, physical examinations and visual acuity examinations. The χ 2 test and binary Logistic regression model were used to analyze group differences in the comorbidity of myopia, obesity and depression symptoms and factors influencing the comorbidity. Stratified analysis was applied to analyze the associations between health risk behaviors and the comorbidity.
Results:
The detection rate of comorbidity of myopia, obesity, and depression symptoms among middle school students in Beijing was 3.35%, the comorbidity rate among vocational high school students (4.61%) was higher than that in junior high school students (2.80%) and senior high school students (3.41%). The comorbidity rate was higher among students in suburban areas (3.66%) than that in urban areas (2.92%), and the differences was statistically significant ( χ 2=15.02, 4.63, P <0.05). Binary Logistic regression analyses indicated that middle school students with poor dietary behaviors ( OR =1.59) and excessive screen time ( OR =1.70) were associated with elevated risk of comorbidity of myopia, obesity, and depression symptoms. Both boys and girls with poor dietary behaviors ( OR =1.63, 1.69) and excessive screen time ( OR =1.45, 2.23) had elevated likelihood of comorbidity of myopia, obesity and depression symptoms. Students in junior high school and senior high school with poor dietary behaviors ( OR =2.16, 1.47) and excessive screen time ( OR =2.20, 1.63 ) had elevated likelihood of comorbidity of myopia, obesity, and depression symptoms ( P <0.05).
Conclusions
The current status of comorbidity of myopia, obesity, and depression symptoms among middle school students in Beijing is concerning. Schools and parents should work together to guide students to develop healthy behaviors such as balanced diet and moderate video, in order to achieve the goal of controlling myopia, obesity and depression symptoms.
5.Cognitive function in patients with early onset and adult onset schizophrenia
Bingjie HUANG ; Jiaheng XIE ; Chengcheng PU ; Huining GUO ; Lei YANG ; Xue HAN ; Zhang CHENG ; Yanbo YUAN ; Jingping ZHAO ; Chuanyue WANG ; Zheng LU ; Fude YANG ; Hong DENG ; Chuan SHI ; Xin YU
Chinese Mental Health Journal 2019;33(3):161-166
Objective:To explore the characteristics of cognitive function in patients with early onset and adult onset schizophrenia.Methods:In this cross-sectional study, 546 patients with schizophrenia who met the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-Ⅳ) were selected.Among them, 62 cases were defined as early onset schizophrenia (EOS, age of onset<18 years) and 175 patients were defined as adult onset schizophrenia (AOS, age of onset≥25 years).Patients underwent clinical assessments with the Positive and Negative Symptom Scale (PANSS) and the Personal and Social Performance Scale (PSP), and comprehensive neuropsychological assessments.Results:The EOS patients got lower scores in motor function-PEGDOM T score [ (26±12) vs. (30±11), P<0.01], working memory-average T score of PASAT and WMSSP[ (34±12) vs. (38±10), P<0.05]and executive function (inhibition) -Stroop T score [ (35±12) vs. (39±10), P<0.05]than AOS patients.No differences were fund in processing speed, verbal memory and learning, visual memory and learning (Ps>0.05) between the two groups.Conclusion:It suggests that the EOS patients have worse motor function, working memory and inhibition.
6.The effect of the ratio of aneurysm sac diameter to patient age on the long-term efficacy of different surgical methods for infrarenal abdominal aortic aneurysm
Ren LIN ; Songbiao ZHAN ; Jiesheng QIAN ; Haipeng HE ; Yang ZHAO ; Junbing LYU ; Jiaxin PENG ; Yibo ZHANG ; Huining CHEN ; Henghui YIN
Chinese Journal of Surgery 2024;62(6):598-605
Objective:To investigate the effect of the ratio of the maximum diameter of aneurysm sac to age (R) on the long-term efficacy of endovascular aneurysm repair (EVAR) and open surgical repair (OSR) in patients with infrarenal abdominal aortic aneurysm (IAAA).Methods:This is a retrospective cohort study.The clinical data of 317 patients with IAAA who underwent surgical repair in the Department of Vascular Surgery,the Third Affiliated Hospital of Sun Yat-Sen University from January 2016 to October 2022 were retrospectively collected.There were 266 males and 51 females,aged (69.7±8.3) years (range:37 to 87 years).The R value of the patient was calculated and the receiver operating characteristic(ROC) curve was used to establish a model to calculate the optimal cut-off value.The propensity score matching method was used to match the baseline data of patients in the EVAR and OSR group by 3∶1 (the caliper value was 0.05),and the patients were stratified according to the cutoff value of R, and the postoperative efficacy and survival of the patients were analyzed.The primary endpoint was the total mortality rate,and the secondary endpoints included the occurrence of postoperative complications and reintervention.Pearson χ2 or Fisher ′s exact test was used for categorical variables, and independent sample t test or Wilcoxon rank sum test was used for continuous variables to compare differences between groups.The survival curves of the two groups were described by Kaplan-Meier method. Results:After propensity score matching,198 cases were in the EVAR group and 66 cases were in the OSR group.The ROC model showed that the best cut-off value of R value was 0.90,and the two groups were divided into two layers:R<0.90 and R≥0.90.Among them,112 patients with R<0.90 (84 cases of EVAR,28 cases of OSR);there were 152 patients with R≥0.90 (114 cases of EVAR and 38 cases of OSR).The follow-up time was (23.6±1.6) months (range:1 to 70 months).In the R≥0.90 stratification,the total mortality (26.3% vs.5.3%, χ2=7.600, P=0.006),complication rate (44.7% vs.26.3%, χ2=4.025, P=0.045), and secondary intervention rate (31.6% vs.13.2%, χ2=4.910, P=0.027) in the EVAR group were higher than those in the OSR group.In the R<0.90 stratification,there was no significant difference in the total mortality rate (13.1% vs.10.7%, χ2=0.109, P=0.741), complication rate (28.6% vs.35.7%, χ2=0.507, P=0.477) and secondary intervention rate (14.3% vs.21.4%, χ2=0.353, P=0.552) between the two groups. Conclusions:When R≥0.90 in IAAA patients,OSR maybe more beneficial to patients in terms of survival rate,postoperative complication rate and secondary intervention rate than EVAR.When R<0.90,there are no significant differences in survival rate,complication rate and secondary intervention rate between the two surgical methods.
7.The effect of the ratio of aneurysm sac diameter to patient age on the long-term efficacy of different surgical methods for infrarenal abdominal aortic aneurysm
Ren LIN ; Songbiao ZHAN ; Jiesheng QIAN ; Haipeng HE ; Yang ZHAO ; Junbing LYU ; Jiaxin PENG ; Yibo ZHANG ; Huining CHEN ; Henghui YIN
Chinese Journal of Surgery 2024;62(6):598-605
Objective:To investigate the effect of the ratio of the maximum diameter of aneurysm sac to age (R) on the long-term efficacy of endovascular aneurysm repair (EVAR) and open surgical repair (OSR) in patients with infrarenal abdominal aortic aneurysm (IAAA).Methods:This is a retrospective cohort study.The clinical data of 317 patients with IAAA who underwent surgical repair in the Department of Vascular Surgery,the Third Affiliated Hospital of Sun Yat-Sen University from January 2016 to October 2022 were retrospectively collected.There were 266 males and 51 females,aged (69.7±8.3) years (range:37 to 87 years).The R value of the patient was calculated and the receiver operating characteristic(ROC) curve was used to establish a model to calculate the optimal cut-off value.The propensity score matching method was used to match the baseline data of patients in the EVAR and OSR group by 3∶1 (the caliper value was 0.05),and the patients were stratified according to the cutoff value of R, and the postoperative efficacy and survival of the patients were analyzed.The primary endpoint was the total mortality rate,and the secondary endpoints included the occurrence of postoperative complications and reintervention.Pearson χ2 or Fisher ′s exact test was used for categorical variables, and independent sample t test or Wilcoxon rank sum test was used for continuous variables to compare differences between groups.The survival curves of the two groups were described by Kaplan-Meier method. Results:After propensity score matching,198 cases were in the EVAR group and 66 cases were in the OSR group.The ROC model showed that the best cut-off value of R value was 0.90,and the two groups were divided into two layers:R<0.90 and R≥0.90.Among them,112 patients with R<0.90 (84 cases of EVAR,28 cases of OSR);there were 152 patients with R≥0.90 (114 cases of EVAR and 38 cases of OSR).The follow-up time was (23.6±1.6) months (range:1 to 70 months).In the R≥0.90 stratification,the total mortality (26.3% vs.5.3%, χ2=7.600, P=0.006),complication rate (44.7% vs.26.3%, χ2=4.025, P=0.045), and secondary intervention rate (31.6% vs.13.2%, χ2=4.910, P=0.027) in the EVAR group were higher than those in the OSR group.In the R<0.90 stratification,there was no significant difference in the total mortality rate (13.1% vs.10.7%, χ2=0.109, P=0.741), complication rate (28.6% vs.35.7%, χ2=0.507, P=0.477) and secondary intervention rate (14.3% vs.21.4%, χ2=0.353, P=0.552) between the two groups. Conclusions:When R≥0.90 in IAAA patients,OSR maybe more beneficial to patients in terms of survival rate,postoperative complication rate and secondary intervention rate than EVAR.When R<0.90,there are no significant differences in survival rate,complication rate and secondary intervention rate between the two surgical methods.
8.Incidence of postoperative complications in Chinese patients with gastric or colorectal cancer based on a national, multicenter, prospective, cohort study
Shuqin ZHANG ; Zhouqiao WU ; Bowen HUO ; Huining XU ; Kang ZHAO ; Changqing JING ; Fenglin LIU ; Jiang YU ; Zhengrong LI ; Jian ZHANG ; Lu ZANG ; Hankun HAO ; Chaohui ZHENG ; Yong LI ; Lin FAN ; Hua HUANG ; Pin LIANG ; Bin WU ; Jiaming ZHU ; Zhaojian NIU ; Linghua ZHU ; Wu SONG ; Jun YOU ; Su YAN ; Ziyu LI
Chinese Journal of Gastrointestinal Surgery 2024;27(3):247-260
Objective:To investigate the incidence of postoperative complications in Chinese patients with gastric or colorectal cancer, and to evaluate the risk factors for postoperative complications.Methods:This was a national, multicenter, prospective, registry-based, cohort study of data obtained from the database of the Prevalence of Abdominal Complications After Gastro- enterological Surgery (PACAGE) study sponsored by the China Gastrointestinal Cancer Surgical Union. The PACAGE database prospectively collected general demographic characteristics, protocols for perioperative treatment, and variables associated with postoperative complications in patients treated for gastric or colorectal cancer in 20 medical centers from December 2018 to December 2020. The patients were grouped according to the presence or absence of postoperative complications. Postoperative complications were categorized and graded in accordance with the expert consensus on postoperative complications in gastrointestinal oncology surgery and Clavien-Dindo grading criteria. The incidence of postoperative complications of different grades are presented as bar charts. Independent risk factors for occurrence of postoperative complications were identified by multifactorial unconditional logistic regression.Results:The study cohort comprised 3926 patients with gastric or colorectal cancer, 657 (16.7%) of whom had a total of 876 postoperative complications. Serious complications (Grade III and above) occurred in 4.0% of patients (156/3926). The rate of Grade V complications was 0.2% (7/3926). The cohort included 2271 patients with gastric cancer with a postoperative complication rate of 18.1% (412/2271) and serious complication rate of 4.7% (106/2271); and 1655 with colorectal cancer, with a postoperative complication rate of 14.8% (245/1655) and serious complication rate of 3.0% (50/1655). The incidences of anastomotic leakage in patients with gastric and colorectal cancer were 3.3% (74/2271) and 3.4% (56/1655), respectively. Abdominal infection was the most frequently occurring complication, accounting for 28.7% (164/572) and 39.5% (120/304) of postoperative complications in patients with gastric and colorectal cancer, respectively. The most frequently occurring grade of postoperative complication was Grade II, accounting for 65.4% (374/572) and 56.6% (172/304) of complications in patients with gastric and colorectal cancers, respectively. Multifactorial analysis identified (1) the following independent risk factors for postoperative complications in patients in the gastric cancer group: preoperative comorbidities (OR=2.54, 95%CI: 1.51-4.28, P<0.001), neoadjuvant therapy (OR=1.42, 95%CI:1.06-1.89, P=0.020), high American Society of Anesthesiologists (ASA) scores (ASA score 2 points:OR=1.60, 95% CI: 1.23-2.07, P<0.001, ASA score ≥3 points:OR=0.43, 95% CI: 0.25-0.73, P=0.002), operative time >180 minutes (OR=1.81, 95% CI: 1.42-2.31, P<0.001), intraoperative bleeding >50 mL (OR=1.29,95%CI: 1.01-1.63, P=0.038), and distal gastrectomy compared with total gastrectomy (OR=0.65,95%CI: 0.51-0.83, P<0.001); and (2) the following independent risk factors for postoperative complications in patients in the colorectal cancer group: female (OR=0.60, 95%CI: 0.44-0.80, P<0.001), preoperative comorbidities (OR=2.73, 95%CI: 1.25-5.99, P=0.030), neoadjuvant therapy (OR=1.83, 95%CI:1.23-2.72, P=0.008), laparoscopic surgery (OR=0.47, 95%CI: 0.30-0.72, P=0.022), and abdominoperineal resection compared with low anterior resection (OR=2.74, 95%CI: 1.71-4.41, P<0.001). Conclusion:Postoperative complications associated with various types of infection were the most frequent complications in patients with gastric or colorectal cancer. Although the risk factors for postoperative complications differed between patients with gastric cancer and those with colorectal cancer, the presence of preoperative comorbidities, administration of neoadjuvant therapy, and extent of surgical resection, were the commonest factors associated with postoperative complications in patients of both categories.
9.Incidence of postoperative complications in Chinese patients with gastric or colorectal cancer based on a national, multicenter, prospective, cohort study
Shuqin ZHANG ; Zhouqiao WU ; Bowen HUO ; Huining XU ; Kang ZHAO ; Changqing JING ; Fenglin LIU ; Jiang YU ; Zhengrong LI ; Jian ZHANG ; Lu ZANG ; Hankun HAO ; Chaohui ZHENG ; Yong LI ; Lin FAN ; Hua HUANG ; Pin LIANG ; Bin WU ; Jiaming ZHU ; Zhaojian NIU ; Linghua ZHU ; Wu SONG ; Jun YOU ; Su YAN ; Ziyu LI
Chinese Journal of Gastrointestinal Surgery 2024;27(3):247-260
Objective:To investigate the incidence of postoperative complications in Chinese patients with gastric or colorectal cancer, and to evaluate the risk factors for postoperative complications.Methods:This was a national, multicenter, prospective, registry-based, cohort study of data obtained from the database of the Prevalence of Abdominal Complications After Gastro- enterological Surgery (PACAGE) study sponsored by the China Gastrointestinal Cancer Surgical Union. The PACAGE database prospectively collected general demographic characteristics, protocols for perioperative treatment, and variables associated with postoperative complications in patients treated for gastric or colorectal cancer in 20 medical centers from December 2018 to December 2020. The patients were grouped according to the presence or absence of postoperative complications. Postoperative complications were categorized and graded in accordance with the expert consensus on postoperative complications in gastrointestinal oncology surgery and Clavien-Dindo grading criteria. The incidence of postoperative complications of different grades are presented as bar charts. Independent risk factors for occurrence of postoperative complications were identified by multifactorial unconditional logistic regression.Results:The study cohort comprised 3926 patients with gastric or colorectal cancer, 657 (16.7%) of whom had a total of 876 postoperative complications. Serious complications (Grade III and above) occurred in 4.0% of patients (156/3926). The rate of Grade V complications was 0.2% (7/3926). The cohort included 2271 patients with gastric cancer with a postoperative complication rate of 18.1% (412/2271) and serious complication rate of 4.7% (106/2271); and 1655 with colorectal cancer, with a postoperative complication rate of 14.8% (245/1655) and serious complication rate of 3.0% (50/1655). The incidences of anastomotic leakage in patients with gastric and colorectal cancer were 3.3% (74/2271) and 3.4% (56/1655), respectively. Abdominal infection was the most frequently occurring complication, accounting for 28.7% (164/572) and 39.5% (120/304) of postoperative complications in patients with gastric and colorectal cancer, respectively. The most frequently occurring grade of postoperative complication was Grade II, accounting for 65.4% (374/572) and 56.6% (172/304) of complications in patients with gastric and colorectal cancers, respectively. Multifactorial analysis identified (1) the following independent risk factors for postoperative complications in patients in the gastric cancer group: preoperative comorbidities (OR=2.54, 95%CI: 1.51-4.28, P<0.001), neoadjuvant therapy (OR=1.42, 95%CI:1.06-1.89, P=0.020), high American Society of Anesthesiologists (ASA) scores (ASA score 2 points:OR=1.60, 95% CI: 1.23-2.07, P<0.001, ASA score ≥3 points:OR=0.43, 95% CI: 0.25-0.73, P=0.002), operative time >180 minutes (OR=1.81, 95% CI: 1.42-2.31, P<0.001), intraoperative bleeding >50 mL (OR=1.29,95%CI: 1.01-1.63, P=0.038), and distal gastrectomy compared with total gastrectomy (OR=0.65,95%CI: 0.51-0.83, P<0.001); and (2) the following independent risk factors for postoperative complications in patients in the colorectal cancer group: female (OR=0.60, 95%CI: 0.44-0.80, P<0.001), preoperative comorbidities (OR=2.73, 95%CI: 1.25-5.99, P=0.030), neoadjuvant therapy (OR=1.83, 95%CI:1.23-2.72, P=0.008), laparoscopic surgery (OR=0.47, 95%CI: 0.30-0.72, P=0.022), and abdominoperineal resection compared with low anterior resection (OR=2.74, 95%CI: 1.71-4.41, P<0.001). Conclusion:Postoperative complications associated with various types of infection were the most frequent complications in patients with gastric or colorectal cancer. Although the risk factors for postoperative complications differed between patients with gastric cancer and those with colorectal cancer, the presence of preoperative comorbidities, administration of neoadjuvant therapy, and extent of surgical resection, were the commonest factors associated with postoperative complications in patients of both categories.
10.Killing effect of NK92 cells modified with CD33-CAR on CD33+ acute myeloid leukemia cells
LIU Yanzhong1 ; PAN Lijuan1 ; TANG Qulai1 ; SHI Jiangzhou1 ; ZHAO Wenjing1 ; HUO Lihong1 ; GU Chaojiang2 ; HU Guang2 ; LIU Huining ; ZHANG Tongcun
Chinese Journal of Cancer Biotherapy 2018;25(5):462-468
[Abstract] Objective: To construct CD33-CAR modified NK92 cells based on CD33-scFv sequence, and to explore its killing effect on CD33+ AML (acute myeloid leukemia) cells. Methods: DNA fragment encoding CD33-CAR was synthesized by gene synthesis and molecular cloning technology and then cloned into lentiviral vector. Lentivirus were packaged and used to transfect NK92 cells. The transfection efficiency was detected by flow cytometry, and puromycin was used to screen NK92 cells stably expressing CD33-CAR (CD33-CAR-NK92). Killing effect of CD33-CAR-NK92 cells on AML cells in vitro was examined with calcein-AM release assays. IFN-γ secretions of NK92 cells and CD33-CAR-NK92 cells were measured by ELISA. Results: The pCDH-CD33-CAR lentiviral vector was successfully constructed. After lentiviral transfection, about 18.7% of NK92 cells express CD33-CAR (referred as CD33-CARNK92 cells). The percentage of CD33-CAR+ NK92 cells was about 86.3% after puromycin selection. In contrast to unmodified NK92 cells, significantly higher cytotoxic effect against CD33+ MOLM-13 cells was found in CD33-CAR-NK92 cells (P<0.01); however, there was no significant difference in cytotoxicity against CD33- JURKAT cells between NK92 cells and CD33-CAR-NK92 cells (P> 0.05). After co-culture at an effect-target ratio of 2∶1 for 6 hours, the level of IFN-γ secreted by the CD33-CAR modified NK92 cells was significantly higher than that of the unmodified ([190.97±11.52] vs [88.41±2.75]pg/ml, P<0.01). Conclusion: The CD33-CARNK92 cells could specifically recognize CD33 antigen and kill CD33+ AML cells in comparison with the unmodified NK92 cells, which provides experimental basis for clinical transformation of CD33-CAR-NK92 cells in treatingAML.