1.Preparation of CD52-targeted chimeric antigen receptor-modified T cells and their anti-leukemia effects.
Yan LIU ; Yu LIU ; Ke Jing TANG ; Zhao Qi CHEN ; Jun Li MOU ; Ying Xi XU ; Hai Yan XING ; Zheng TIAN ; Qing RAO ; Min WANG ; Jian Xiang WANG
Chinese Journal of Hematology 2022;43(4):279-286
Objective: To construct chimeric antigen receptor (CAR) T cells targeting CD52 (CD52 CAR-T) and validate the effect of CD52 CAR-T cells on CD52-positive leukemia. Methods: A second-generation CD52-targeting CAR bearing 4-1BB costimulatory domain was ligated into a lentiviral vector through molecular cloning. Lentivirus was prepared and packaged by 293 T cells with a four-plasmid system. Fluorescein was used to label cell surface antigens to evaluate the phenotype of CD52 CAR-T cells after infection. Flow cytometry and ELISA were used to evaluate the specific cytotoxicity of CD52 CAR-T cells to CD52-positive cell lines in vitro. Results: ①A pCDH-CD52scFv-CD8α-4-1BB-CD3ζ-GFP expressing plasmid was successfully constructed and used to transduce T cells expressing a novel CD52-targeting CAR. ②On day 6, CD52-positive T cells were almost killed by CD52-targeted CAR-T post lentivirus transduction [CD52 CAR-T (4.48 ± 4.99) %, vs Vector-T (56.58±19.8) %, P=0.011]. ③T cells transduced with the CAR targeting CD52 showed low levels of apoptosis and could be expanded long-term ex vivo. ④The CD52 CAR could promote T cell differentiation into central and effector memory T cells, whereas the proportion of T cells with a CD45RA(+) effector memory phenotype were reduced. ⑤CD52 CAR-T cells could specifically kill CD52-positive HuT78-19t cells but had no killing effect on CD52-negative MOLT4-19t cells. For CD52 CAR-T cells, the percentage of residual of HuT78-19t cells was (2.66±1.60) % at an the E:T ratio of 1∶1 for 24 h, while (56.66±5.74) % of MOLT4-19t cells survived (P<0.001) . ⑥The results of a degranulation experiment confirmed that HuT78-19t cells significantly activated CD52 CAR-T cells but not MOLT4-19t cells[ (57.34±11.25) % vs (13.06± 4.23) %, P<0.001]. ⑦CD52 CAR-T cells released more cytokines when co-cultured with HuT78-19t cells than that of vector-T cells [IFN-γ: (3706±226) pg/ml, P<0.001; TNF-α: (1732±560) pg/ml, P<0.01]. Conclusions: We successfully prepared CD52 CAR-T cells with anti-leukemia effects, which might provide the foundation for further immunotherapy.
CD52 Antigen
;
Cell Line, Tumor
;
Humans
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Immunotherapy, Adoptive/methods*
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Lentivirus/genetics*
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Leukemia
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Receptors, Antigen, T-Cell
;
Receptors, Chimeric Antigen/genetics*
2.Analysis of Influence of Jiangxi Characteristic Processing Technology on Chemical Compositions of Aurantii Fructus Based on UPLC-Q/TOF-MS
Jing ZHU ; En YUAN ; Jun-yong MOU ; Ling-yun ZHONG ; Jin-lian ZHANG ; Ming YANG ; Hai-ping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(16):142-153
Objective:To compare the effects of different processing techniques on the chemical constituents of Aurantii Fructus for screening the dominant decoction pieces. Method:UPLC-Q/TOF-MS was used to detect the chemical constituents of Aurantii Fructus, chromatography separation was achieved on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm), and gradient elution was performed with 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) as mobile phase (0-10 min, 5%-35%B; 10-18 min, 35%-75%B; 18-21 min, 75%-100%B; 21-24 min, 100%B; 24-24.1 min, 100%-5%B; 24.1-28 min, 5%B). Data acquisition was carried out in electrospray ionization (ESI) under the positive ion mode, the scanning range was
3.Application of endoscopic injection of lauromacrogol and methylene blue in sclerotherapy of esophageal varices
Hai-Jun MOU ; Jing-Yu XU ; Bi-Guang TUO ; Yuan GAO ; Yuan YANG ; Xiao-Lin ZHOU
China Journal of Endoscopy 2018;24(3):1-5
Objective To discuss the application of endoscopic injection of lauromacrogol and methylene blue in sclerotherapy of esophageal varices. Methods The clinical data of 62 patients with esophageal varices underwent endoscopic treatment from June 2014 to February 2016 were collected, including 26 cases treated by endoscopic injection of lauromacrogol (A group) and 36 cases treated by endoscopic injection of lauromacrogol and methylene blue (B group). The treatment effects, success rate of hemostasis, safety, complications and follow-up recurrence were compared between the two groups. Results The total effective rate and success rate of emergency hemostasis in B group were slightly higher than those in A group (P > 0.05). The times of treatment for eliminating esophageal varices in B group was less than that in A group, and the duration of treatment was shorter than that in A group (P < 0.05). The long-term rebleeding rate and total bleeding rate in B group were lower than those in A group (P < 0.05). The incidence of postoperative complications and recurrence rate in B group were lower than those in A group (P < 0.05). Conclusion Endoscopic injection of lauromacrogol and methylene blue is effective in the treatment of esophageal varices, and the hemostatic effect is good. The incidence of postoperative complications and the recurrence rate are low.
4.Clinical Effect Observation of Jianpishugan and Huoxuehuatan Formula on Liver Depression and Spleen Deficiency Induced Non-Alcoholic Steatohepatitis
Yong HU ; Hai-Jun MOU ; Ke CHEN
Journal of Nanjing University of Traditional Chinese Medicine 2017;33(4):359-362
OBJECTIVE To study the clinical effect of Jianpishugan and Huoxuehuatan formula on liver depression and spleen deficiency induced non-alcoholic steatohepatitis.METHODS 140 patients with liver depression and spleen deficiency induced non-alcoholic steatohepatitis admitted in our hospital from Oct.2015 to Oct.2016 were selected and randomly divided into the observation group and the control group according to the random number.Both groups were given exercise and diet therapy,and the control group was also treated with polyene phosphatidylcholine capsules.Based on the control group,the observation group was also treated with modified Jianpishugan and Huoxuehuatan Formula.RESULTS Symptom scores and the total scores of the clinical symptoms in two groups decreased significantly (P<0.01) after the treatment,but that in the observation group decreased more significantly (P<0.01).Total effective rate of overall efficacy in the observation group was 94.28% after the treatment,which was significantly higher than that in the control group (77.14%,P<0.01).Total effective rate of TCM syndrome in the observation group was 95.71% after the treatment,which was significantly higher than that in the control group (77.14%,P<0.01).ALT,AST and GGT levels in two groups decreased significantly (P<0.01) after the treatment,but that in the observation group decreased more significantly (P<0.01).Abdominal B ultrasound results in two groups improved significantly after the treatment(P<0.01),but that in the observation group improved more significantly (P<0.01).During the treatment,patients in two groups did not complain of other discomforts,adverse reactions and toxic effects.CONCLUSION Jianpishugan and Huoxuehuatan Formula can noticeably improve the clinical symptoms and liver functions of patients with liver depression and spleen deficiency induced non-alcoholic steatohepatitis.
5.Risk factors associated with postoperative complications of D2 radical resection for local advanced gastric cancer.
Xiao-liang LAN ; Guo-xin LI ; Hao LIU ; Yan-feng HU ; Hai-jun DENG ; Ting-yu MOU
Chinese Journal of Gastrointestinal Surgery 2013;16(8):764-767
OBJECTIVETo evaluate the risk factors of postoperative complications following D2 radical resection for advanced gastric cancer.
METHODSFrom June 2004 to May 2011, 483 patients with local advanced gastric cancer who underwent radical gastrectomy with D2 lymph node dissection were enrolled in the study, including 132 patients of LAG (27.3%) and 351 patients of open procedure (72.7%). Clinicopathological data and postoperative complications were reviewed retrospectively. Postoperative complications were classified into overall and severe complications according to Clavien-Dindo Classification. Multivariate logistic model was used to identify risk factors of postoperative complications.
RESULTSThe overall incidence of postoperative overall and severe complications and mortality were 12.4% (60/483), 2.5% (12/483) and 0.2% (1/483), respectively. Univariate analysis showed that no significant differences were found in overall and severe complications between the two surgical approaches (13.6% vs. 12.0%, P=0.620; 3.0% vs. 2.3%, P=0.743). Furthermore, multivariate analysis showed that age ≥60 years, preoperative comorbidity and intraoperative blood loss >300 ml were independent risk factors associated with overall postoperative complications. Remarkably, intraoperative blood loss >300 ml was also an independent risk factor for severe postoperative complications.
CONCLUSIONSLAG with D2 lymph node dissection for local advanced gastric cancer is technically feasible and safe. However, the elderly, preoperative comorbidity and increased intraoperative blood loss are associated with elevated risk of complications. Decreased intraoperative bleeding may reduce the potential postoperative complications.
Female ; Gastrectomy ; Humans ; Laparoscopy ; Laparotomy ; Logistic Models ; Lymph Node Excision ; Male ; Middle Aged ; Multivariate Analysis ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; surgery
6.Study on force feedback of acupuncture at Fengchi (GB 20).
Jun JIANG ; Fu-Bo WANG ; Hai-Dong GUO ; Shui-Jin SHAO ; Peng MIAO ; Qi ZHANG ; Chun-Xia GUO ; Fang-Fang MOU ; Zhen-Guo YAN
Chinese Acupuncture & Moxibustion 2013;33(10):939-942
Acupuncture manipulations on Fengchi (GB 20) of famous doctors were taken through force feedback device, then the data was input into a digitized virtual human. Virtual Fengchi (GB 20) acupuncture force feedback simulation system was built through the virtual reality technology to achieve one-to-one high simulative manipulation effect for acupuncture students. The interaction force of the needle body and human tissues was analyzed during the acupuncture process on the 3D digital human integrated with information of Fengchi (GB 20) according to the physical characteristics of the tissues under this point. The mechanical model which is used to imitate the stress received by the body of the needle was established, and transmitted truly to the operator by the force feedback device. Thus, Fengchi (GB 20) virtual acupuncture force feedback simulation was preliminary established, and the sense of touch could be reproduced lively on the visualized virtual acupuncture human. It is held that Fengchi (GB 20) acupuncture force feedback research is a preliminary exploration for virtual acupuncture that integrated with the information of visual, tactile and force feedback. And it also provided a dynamic one-to-one simulation approach for acupuncture teaching.
Acupuncture
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education
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Acupuncture Points
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Acupuncture Therapy
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instrumentation
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methods
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Computer Simulation
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Humans
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Needles
7.Trichloroethylene induces biphasic concentration-dependent changes in cell proliferation and the expression of SET-associated proteins in human hepatic L-02 cells.
Wen Xu HONG ; Jin Bo YE ; Mou Tong CHEN ; Yan YAN ; Gui Feng ZHOU ; Xi Fei YANG ; Liang YANG ; Xiao Hu REN ; Hai Yan HUANG ; Li ZHOU ; Xin Feng HUANG ; Zhi Xiong ZHUANG ; Jian Jun LIU
Biomedical and Environmental Sciences 2013;26(7):618-621
Cell Line
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Cell Proliferation
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drug effects
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Dose-Response Relationship, Drug
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Histone Chaperones
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metabolism
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Humans
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Liver
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cytology
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Protein Interaction Maps
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Solvents
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administration & dosage
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toxicity
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Transcription Factors
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metabolism
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Trichloroethylene
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administration & dosage
;
toxicity
8.The safety and long-term survival after laparoscopic surgery versus open surgery for colon cancer.
Li-ying ZHAO ; Ya-nan WANG ; Jiang YU ; Hai-jun DENG ; Yan-feng HU ; Ting-yu MOU ; Guo-xin LI
Chinese Journal of Surgery 2013;51(4):314-319
OBJECTIVETo evaluate the short-term outcomes and 5-year recurrence, overall survival, and disease-free survival of laparoscopic assisted surgery for colon cancer.
METHODSThe clinical and pathologic data were compared between the patients who underwent colectomy during March 2003 to July 2008 and assigned in laparoscopic group (n = 92) and open group (n = 285) according the surgical approach. The 5-year overall survival, disease-free survival, and recurrence rate were analyzed for all patients who were followed-up for more than 36 months in either of the groups.
RESULTSThe laparoscopic colectomy was associated with manifested less blood loss (50(50) ml) (Z = -8.292, P < 0.01), early return of bowel function (the evacuation time was (3.0 ± 1.0) days, and the meal time after operation was (4.0 ± 1.3) days) (t = -6.475 and -4.871, P < 0.01), and longer length (cm) of distal resection margin ((10 ± 4) cm vs. (9 ± 4) cm, t = 3.527, P = 0.000). The 5-year overall survival of the laparoscopic group and the open group were 63.6% and 61.8% respectively. The 5-year disease-free survival of the I-III stage patients in the laparoscopic group and the open group were 69.5% and 65.5% respectively, and the local recurrence were 8.7% and 13.6% (all P > 0.05).
CONCLUSIONThe laparoscopic colectomy for colon cancer is safe in short-term clinical results and non-inferior to the open colectomy in long-term oncological outcomes.
Adult ; Aged ; Colectomy ; methods ; Colonic Neoplasms ; mortality ; surgery ; Female ; Humans ; Laparoscopy ; Laparotomy ; Length of Stay ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Survival Rate ; Treatment Outcome
9.Clinical analysis of anti glomerular basement membrane disease with normal renal function.
Li-jun MOU ; Li-meng CHEN ; Lai-meng ZUO ; Hai-yun WANG ; Jian-ling TAO ; Yu-bing WEN ; Wen--ling YE ; Yan QIN ; Hang LI ; Xue-mei LI ; Xue-wang LI
Acta Academiae Medicinae Sinicae 2011;33(4):432-435
OBJECTIVETo investigate the clinical and pathologic characteristics of anti-glomerular basement membrane(GBM) disease with normal renal function.
METHODSThe clinical and pathologic data of 6 patients with anti-GBM disease and normal renal function in Peking Union Medical College Hospital were reviewed retrospectively. Furthermore, 29 patients with anti-GBM disease and impaired renal function in the same period in the same hospital were enrolled as the control group. Factors that may influence the prognosis were analyzed.
RESULTSSix (17.1%) of all 35 patients maintained normal renal function for 12-133 months during follow-up. Five patients had microhematuria and proteinuria, one had pulmonary hemorrhage only, and three manifested as Goodpasture syndrome. Renal biopsies from 4 patients revealed linear deposition of IgG 2+-3+ along the glomerular capillary walls by immunofluorescence. As shown by normal light microscopy, mild mesangial proliferation and crescentic glomerulonephritis with a large amount of fibrinoid necrosis of glomerular capillary walls were observed in different patients; however, most pathological changes were mild. Five of these six patients were treated with immunosuppressive drugs and/or plasma exchange. Compared with the control group, the 6 patients with normal renal function had significantly higher hemoglobin[(77.97±20.62 vs.(99.67±19.80 g/L P=0.024], lower titers of anti-GBM antibody[(224.34 ± 145.79 vs.(80.23 ± 85.73 EU/ml P=0.027], and lower ratio of glomeruli with crescents[(0.58±0.29 vs.(0.17±0.27 ,P=0.005]. These 6 patients with normal renal function were followed up for 12-133 months, among whom 4 patients achieved complete remission and 2 had mild proteinuria and microhematuria.
CONCLUSIONAnti-GBM disease with normal renal function is not uncommon. Most patients have mild pathologic changes and good prognosis.
Adult ; Anti-Glomerular Basement Membrane Disease ; pathology ; physiopathology ; Female ; Follow-Up Studies ; Humans ; Kidney ; physiopathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies
10.Method improvement of total quality control of the rescue vehicles
Qing-Li WANG ; Hong QU ; Ke-Lan XIANG ; Qiu-Jiang XIA ; Yi-Yun ZHOU ; Qiong-Ya WU ; Hai-Yan XU ; Zong-Jun MOU
Chinese Journal of Modern Nursing 2011;17(8):942-944
Objective Through the unified and standardized management of the rescue vehicle,to ensure rescue items complete integrity, fast and effective.Methods Standard configuration in emergency medicine,placement,storage,registration of all aspects evaluate were unified Results The time for rescuing and counting stuffs,satisfied degree of doctors and valid time of medicine were significantly improved after quality control(P<0.01).Conclusions Standard management of the rescue vehicles could ensure medicines fast.safe and effective.It is worthy of clinical application widely.

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