1.Construction of CD138-targeted chimeric antigen receptor- modified T cells and their effect in multiple myeloma therapy
Chengcai GUO ; Yang LU ; Kejing TANG ; Haiyan XING ; Zheng TIAN ; Qing RAO ; Min WANG ; Dongsheng XIONG ; Jianxiang WANG
Chinese Journal of Hematology 2024;45(5):436-444
Objective:To construct a novel chimeric antigen receptor T (CAR-T) cell targeting CD138 and to investigate its cytotoxicity against myeloma cells.Methods:The hybridoma strain that can stably secrete the CD138 monoclonal antibody (mAb) was prepared and obtained through monoclonal antibody screening technology. The hybridoma strain cells were intraperitoneally injected into mice to produce ascites containing monoclonal antibodies, which were then collected and purified to obtain pure CD138 mAb. Further examinations were performed to assess the biological characteristics of CD138 mAb. The variable region sequence of this antibody was amplified through reverse transcription polymerase chain reaction and was used as the antigen recognition domain of CD138 CAR, which was subsequently expressed on the surface of T cells by lentiviral infection. Flow cytometry was employed to assess the phenotype of CD138 CAR-T cells. In vitro cytotoxicity and degranulation assays were performed to evaluate their antitumor effects.Results:① We successfully prepared anti-human CD138 antibody hybridoma cell lines and screened a hybridoma cell strain, 5G2, which could persistently and stably secrete the anti-CD138 antibody. ② The purified CD138 (5G2) mAb can especially recognize CD138 + cells with a binding affinity constant (K D) of 6.011×10 -9 mol/L and showed no significant binding activity with CD138 - cells. ③The variable region sequence of the CD138 (5G2) antibody was obtained using molecular cloning technology, and CD138 (5G2) CAR was successfully constructed and expressed on T cells through lentivirus infection and, concurrently, demonstrated effective binding to recombinant human CD138 protein.④ The proliferation of T cells transduced with the CD138 (5G2) CAR was highly efficient. The phenotype analysis revealed that CD138 (5G2) CAR-T cells exhibited a greater tendency to differentiate into central memory T cells and memory stem T cells, with a reduced proportion of terminally differentiated effector memory subsets. ⑤CD138 (5G2) CAR-T cells demonstrated specific cytotoxicity against CD138 + myeloma cell line H929, whereas CD138 - cell line K562 remained unaffected. The percentage of residual H929 cells was (12.92±8.02) % after co-culturing with CD138 (5G2) CAR-T cells, while (54.25±15.79) % was left in the Vector-T group (E∶T=1∶2; P<0.001). ⑥Results of degranulation assays demonstrated a significant activation of CD138 (5G2) CAR-T cells after co-culture with the H929 cell line, whereas no significant activation was observed in Vector-T cells [ (25.78±3.35) % vs (6.13±1.30) %, P<0.001]. ⑦After co-culturing with CD138 + cells, CD138 (5G2) CAR-T cells exhibited a significant increase in cytokine secretion compared to the Vector-T group [interleukin-2: (1 697.52±599.05) pg/ml vs (5.07±1.17) pg/ml, P<0.001; interferon-γ: (3 312.20±486.38) pg/ml vs (9.28±1.46) pg/ml, P<0.001; and tumor necrosis factor-α: (1 837.43±640.49) pg/ml vs (8.75±1.65) pg/ml, P<0.001]. However, no significant difference was observed in cytokine secretion levels between the two groups after co-culturing with CD138 - cells. Conclusion:This study successfully prepared a novel monoclonal antibody against CD138, and CAR-T cells constructed with the antigen recognition domain derived from this 5G2 mAb demonstrated effective antitumor activity against myeloma cells. This can be used as a new option for the detection of the CD138 antigen and proposes a novel strategy for multiple myeloma immunotherapy.
2.To construct a risk model and study the immune mechanism of genes related to myocardial infarction and cuproptosis based on bioinformatics and single cell sequencing
Xing JU ; Lianqun JIA ; Zhe ZHANG ; Dongsheng WEI ; Jingsheng ZHANG ; Guanlin YANG
Chinese Journal of Immunology 2024;40(11):2247-2256
Objective:Integrating genes and GEO database related cuproptosis chips,to analyze connection between cupropto-sis genes,immune infiltration and myocardial infarction(MI),construct risk prediction model,predict Western medicine and Chi-nese medicine,analyze miRNA-mRNA regulatory network,and to provides a new research direction for the future study of MI-related cuproptosis mechanism and immune infiltration.Methods:By GEO database retrieval of MI related chips,standardized processing and MI-related cuproptosis genes screening were performed,and immunoinfiltration analysis and quantification were performed based on the treated gene expression matrix,correlation between immune infiltrating cells and function was analyzed,as well as their differ-ences in MI group and the control group.Cuproptosis genes that most related to MI in immune infiltration were screened out,and the risk model was constructed to analyze the risk probability of cuproptosis genes in MI.The Enrichr website and Coremine Medical data-base were used to predict cuproptosis-related genes in MI in Western medicine and traditional Chinese medicine.Finally,the up-stream mirnas of FDX1 and SLC31A1 were predicted by miRTarBase,Starbase and Targetscan databases,and miRNA-mRNA regula-tory networks were constructed.Results:Correlation of immune infiltration showed that Tfh cells and B cells had the strongest positive correlation(r=0.68),while regulatory T cells and iDC had the strongest negative correlation(r=-0.63);the difference analysis of im-mune infiltration showed that the differences among mast cells,NK cells and Th1 cells in the MI group at the cellular level were the most significant(P<0.001);and the differences in APC co-inhibition and MHCⅠ at the functional level were the most significant(P<0.001).Six genes with the highest correlation between immune cells and immune function were screened out:ATP7A,DLD,FDX1,LIAS,LIPT1 and SLC31A1.Results of the risk model showed that the high levels of FDX1 and SLC31A1 were negatively correlated with the risk prediction of MI.A total of 21 Western medicines and 30 traditional medicines were predicted by database comparison.miRTarBase,Starbase and Targetscan databases predicted 9 upstream miRNAs of cuproptosis-related genes in MI,including has-miR-122-5p.Conclusion:Tfh cells,B cells,para-inflammatory,typeⅠinterferon response and other related immune cells and functions may play important roles in pathogenesis and prognosis of MI.FDX1 and SLC31A1 as the key genes of cuproptosis process,are negatively correlated with MI.A total of 30 kinds of traditional Chinese medicines including Spirulina,sheep liver,Wen ezhu,Pian jianghuang and Yujin may have potential value in treatment of MI.Finally,9 miRNAs including has-miR-122-5p may play an important role in the regulation of cuproptosis in myocardial infarction.
3.Supplementing rehabilitation training with botulinum neurotoxin improves outcomes for Parkinson′s disease patients with striatal foot deformity
Xue LI ; Liuyi LI ; Shucheng XING ; Siyuan CHEN ; Shaopu WU ; Qi GU ; Dongsheng LI ; Jianjun MA
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(2):146-150
Objective:To observe any therapeutic effect of combining botulinum toxin type A (BTX-A) with rehabilitation training in treating Parkinson′s disease (PD) patients with striatal foot deformity (SFD).Methods:A total of 68 PD patients with SFD were randomly divided into a control group and a treatment group. Both groups were given routine medication with pramipexole and dopamine receptor agonists and received lower limb rehabilitation training, including passive activity training, strength training and walking training. The treatment group was additionally injected with BTX-A. Sciatic pain was quantified using a visual analogue scale. The Unified Parkinson′s Disease Rating Scale-lower limb motor lower limb motor function (UPDRS-LLM) scale, the Berg balance scale and the modified Barthel index were applied to test all of the participants before the experiment and on the 7th, 14th and 30th day of the treatment.Results:The average scores of the control group on all of measures at were significantly better than those of the control group at the same time points, and by the 14th and 30th day had improved significantly compared with those before treatment.Conclusion:Supplementing rehabilitation training with BTX-A can significantly improve foot deformity and relieve the muscle tension and spastic pain of PD patients with SFD, promoting the motor functioning of their lower limbs, their balance and their performance in the activities of daily living.
4.Study on the relationship between the pathological type of lung cancer caused by coke oven discharge and the contact type and exposure time.
Lin Lin WANG ; Xing LIANG ; Ke Jun JIA ; Yong Ping HE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(10):761-764
Objective: To study the pathological types of lung cancer caused by coke oven emissions and analyze the correlation between different exposure levels. Methods: In October 2020, the relevant data of 86 confirmed cases of lung cancer caused by coke oven emissions (including basic information of patients, relevant occupational exposure and clinical data) were collected, The workers were grouped according to the different COEs concentrations in their posts: workers in auxiliary posts were taken as the low exposure group (11 persons) , The workers at coke side and furnace bottom are the medium exposure group (14 persons) , and the workers at furnace top are the high exposure group (61 persons) , and the correlation between pathological types of lung cancer and different exposure levels was analyzed. Results: There was no significant difference in age and length of service among the groups (P>0.05) ; The number of lung cancer cases and pathological types among workers in each group were statistically significant (P=0.044) . After adjusting for interference factors, the number of undifferentiated cancers (mainly small cell lung cancer) increased with the increase of exposure level, with a statistically significant difference (P=0.001) . The incidence of lung cancer increased gradually with the length of service, and the incidence rate of lung cancer among workers of different working ages was statistically significant (P<0.05) . Conclusion: Undifferentiated small cell carcinoma is the most common pathological type of lung cancer caused by coke oven emissions, and the incidence of lung cancer tends to increase with the length of service.
Humans
;
Coke/analysis*
;
Lung Neoplasms/epidemiology*
;
Occupational Exposure/analysis*
;
Polycyclic Aromatic Hydrocarbons/analysis*
;
Pyrenes/analysis*
5.Establishment and Analysis of the Fingerprints of Adiantum capillusveneris from Different Producing Areas and Content Determination of Its Differential Components
Dan XIAO ; Dongsheng FAN ; Xing LIU ; Yi KONG ; Jinjing XUE ; Yuan GAO
China Pharmacy 2021;32(17):2085-2090
OBJECTIVE:To establish the fingerprint of Adiantum capillusveneris from different producing areas ,and to conduct chemometric analysis and content determination of differential components ,so as to provide reference for quality control of A. capillusveneris . METHODS :HPLC-DAD combined with Similarity Evaluation System of TCM Chromatogramtic Fingerprint (2004 A edition )were used to establish fingerprint of 19 batches of A. capillusveneris from different producing areas (S1-S19). Common peaks were confirmed and their similarities were evaluated. Chemometric analysis methods such as cluster analysis , principle component analysis (PCA),orthogonal partial least squares discriminant analysis (OPLS-DA)were used to evaluate the quality of 19 batches of A. capillusveneris from different producing areas ,screen the differential components ,and determine the contents of some differential components. RESULTS :Among 19 batches of A. capillusveneris from different producing areas ,22 common peaks were confirmed ;peak 9 was chlorogenic acid ,peak 17 was quercetin- 3-O-β-D-glucopyranoside,peak 20 was kaempferol-3-O-rutoside;the similarity of 19 batches of sample were 0.677-0.962. Through cluster analysis ,it was found that S 7 and S 10 were clustered into one category ;S15 and S 18 were clustered into one category ;and S 1-S6,S8,S9,S11-S14,S16,S17 and S 19 were clustered into one category. PCA and OPLS-DA found that S 7 and S 10 were clustered into one category ;S15 were clustered into one category ;S18 were clustered into one category ;S1-S6,S8,S9,S11-S14,S16,S17 and S 19 were clustered into one category. Chlorogenic acid ,quercetin-3-O-β-D-glucopyranoside,kaempferol-3-O-rutoside and chemical composition represented by peak 14 were the differential components of the〔2017〕2841); medicinal material. Among 19 batches of A. capillusveneris , average contents of chlorogenic acid and quercetin- 3-O-β-D- glucopyranoside and kaempferol- 3-O-rutoside were 0.10-4.25, 0.31-7.11,0.61-12.00 mg/g,respectively. CONCLUSIONS : 电话:0851-86614212。 HPLC-DAD fingerprints of A. capillusveneris from different producing areas are establishe d in the study ,and three common peaks are identified. Four differential components affecting the quality of A. capillusveneris are screened , and the contents of chlorogenic acid , quercetin-3-O-β-D-glucopyranoside and kaempferol-3-O-rutoside in A. capillusveneris from different producing areas were significantly different.
6.Gadolinium angiography of inner ear in patients with sudden deafness and its clinical analysis.
Yong Ping HAN ; Jun Yi ZHANG ; Lei LIU ; Cai Feng LIANG ; Guo Hua MIAO ; Zheng Bang XU ; Lei WU ; Xue Qin DU ; Wen Jie FAN ; Ying XING ; Li Sheng YU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2021;56(7):746-750
Objective: To observe the gadolinium imaging findings of inner ear in patients with sudden deafness and to analyze its clinical features. Methods: From November 2017 to July 2020, 21 patients with sudden deafness in the People's Hospital of Dongsheng District, Ordos City were selected as the research objects, including 14 males and 7 females, aged 36-76 years, with a median age of 50 years. The course of disease was 1-19 days, with an average of 5.5 days. The patients received audiology tests, laboratory examination, and intravenous gadolinium angiography, each of whom was scanned twice by 3D-FLAIR sequence: once before intravenous gadolinium injection, and once again 4.5-6.0 h after intravenous gadolinium injection. The following corresponding clinical treatment was given. The imaging manifestations and clinical features were observed. Results: Among 21 cases of sudden deafness in acute stage, the signal intensity of 11 cases was significantly higher than that of the contralateral ear, and 2 cases had vestibular labyrinthine hydrops. In laboratory examination, only 2 cases of total deafness had increased WBC count and faster erythrocyte sedimentation rate, and the rest had no abnormality. The hearing types of 21 patients with sudden deafness were: total deafness in 8 cases, flat decline in 10 cases, low frequency decline in 1 case, high frequency decline in 2 cases. The total effective rate was 57% (12/21). The hearing types of 11 patients with abnormal gadolinium angiography were total deafness in 5 cases, flat decline in 5 cases and high frequency decline in 1 case. The total effective rate was 64% (7/11). Conclusion: Gadolinium angiography is abnormal in some patients with sudden deafness, and the permeability of blood labyrinth barrier may be increased, which is worthy of further study.
Angiography
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Deafness
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Female
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Gadolinium
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Hearing Loss, Sudden/diagnostic imaging*
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Humans
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Magnetic Resonance Imaging
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Male
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Middle Aged
;
Vestibule, Labyrinth
7.Effects of Locking Screws with Different Head Angles on Mechanical Properties of Screw-Plate System
Dongsheng JI ; Yun ZHENG ; Erfan HUO ; Hongyan XING
Journal of Medical Biomechanics 2020;35(2):E223-E227
Objective To investigate the effects from head angle and locking times of locking screws on mechanical properties of the screw-plate system, so as to provide a theoretical basis for doctors to select appropriate locking screws and master the locking times in clinical surgical procedure. Methods Locking screws with different head angles were selected to match with locking compression plates in the case of different locking times, and cantilever bending method was used to compare the differences of mechanical properties in each group. Results There were significant differences in failure load and bending stiffness between locking compression plate and locking screw with different head angles (P<0-05). The mechanical properties of the screw-plate system with the best locking degree were optimum. There was no significant difference between 3 time-locking and 1 time-locking (P>0-05). The failure modes of locking screw with different head angles and different locking times were different, and the failure mode of locking screw had a positive correlation with its head angle and locking times. Conclusions The differences in mechanical properties between locking screw with different head angles and locking compression plate cannot be neglected. It is suggested that doctors should choose locking screw and locking compression plate with the best locking degree in clinical procedure, and choose the right torque wrench to lock in multiple times if necessary, thus to prevent screw loosening from affecting recovery of the patients.
8.A preliminary study for special English teaching reform of medical laboratory
Wen LIU ; Dongsheng WANG ; Bin GUO ; Yan LEI ; Linxu ZHAO ; Fang LIN ; Yan XING ; Xiaolan GUO
Chinese Journal of Medical Education Research 2017;16(9):908-911
There are some problems with this course of medical laboratory special English, such as reference materials, teaching contents, teaching ways, the evaluation forms, etc. The department of medical laboratory of North Sichuan Medical College did exploratory reforms including 1+n teaching mode (one major teacher and several co-operational teachers in discussion section), Flipped classroom and interactive teaching, new formative assessment forms (usual performance combing final shows grade), etc. They used innovative teaching way with the purpose of establishing new teaching way , cultivating abilities of au-tonomous learning and comprehensive application.
9.Treatment of comminuted posterior acetabular wall fractures with structural autologous iliac bone graft combined with mini screws
Haomin CUI ; Limeng SUN ; Dongsheng ZHOU ; Lianxin LI ; Xing WANG ; Qinghu LI ; Weicheng XU
Chinese Journal of Orthopaedic Trauma 2016;18(2):126-132
Objective To evaluate the short and mid-term effects of fixing comminuted posterior acetabular wall fractures with structural autologous iliac bone graft combined with mini screws.Methods From January 2010 to January 2014,29 patients with comminuted posterior acetabular wall fracture were treated by structural autologous iliac bone graf combined with mini screws.They were 21 males and 8 females,with a mean age of 44.2 years (range,from 22 to 58 years).The mean time form injury to operation was 7.8 days (range,from 1 to 25 days).The operations were performed through the Kocher-Langenbeck approach,with the patients in the lying position on the uninjured side.The fragments were reduced and fixed by mini screws and the ischemic ones were removed.Structural autologous iliac bone graft was used to reconstruct the posterior wall of acetabulum before a reconstruction plate was applied to compress and maintain it.The functional outcomes were evaluated by the modified Merle d'Aubigne and Postel clinical grading system at the last follow-ups.The radiographs were graded according to the Matta criteria.Results By the Matta criteria,10 cases achieved excellent reduction,16 good reduction,and 3 poor reduction,giving a good to excellent rate of 89.7%.Of this series,29 patients were followed up for 31.5 months on average (range,from 12 to 48 months).By the modified Merle d'Aubigne and Postel criteria,the functional recovery was rated as excellent in 16 cases,good in 9,fair in 3 and poor in one,giving a good to excellent rate of 86.2%.Two cases developed femoral head necrosis according to the magnetic resonance imaging 18 months postoperation.Three patients developed traumatic arthritis two years postoperation.Five patients developed heterotopic ossification postoperation,with no obvious clinical symptoms.Two patient with injury to the sciatic nerve recovered 4 months postoperation.Conclusions Structural autologous iliac graft combined with mini screws can reconstruct the integrity and stability of the fractured acetabular posterior wall,avoiding osteonecrosis of the acetabulum.This surgical technique is effective and safe in treatment of comminuted fracture of the acetabular posterior wall.
10.The Application of DHI Scale to Evaluation of Quality of Life in Patients with Benign Paroxysmal Positional Vertigo
Shiguo XIU ; Dongsheng XING ; Wei HU ; Xin FAN ; Xue ZHANG ; Dongzhu LI
Journal of Audiology and Speech Pathology 2014;(1):48-52
Objective This paper attempts to explore the application of dizziness handicap inventory (DHI) in evaluation of health -related quality of life (QOL ) changes of patients with benign paroxysmal positional vertigo (BPPV) before and after the treatment with canalith repositioning procedure (CRP) .Methods The DHI was em-ployed to investigate and evaluate the dizziness handicap of 120 patients with BPPV before and after 3 months of CRP treatment (treatment group) and 60 healthy controls (control group) ,while the DHI scoring results were com-pared .Results As indicated by DHI evaluation ,the scoring of each DHI items of patients with BPPV before treatment was higher than that of control group ,treatment group before treatment :functional score 22 .60 ± 6 .54 ,emotional score 18 .50 ± 8 .28 ,physical score 17 .90 ± 5 .05 ,total composite score 59 .00 ± 14 .32 .For the control group:functional score 1 .35 ± 1 .74 ,emotional score 1 .00 ± 1 .01 ,physical score 1 .37 ± 1 .86 ,total composite score 3 .72 ± 3 .46 ,with the differ-ence statistically significant (P<0 .05) .The DHI scoring of treatment group of patients with BPPV after 3 months of CRP treatment was lower than that before the treatment (treatment group after 3 months CRP treatment :functional score 1 .10 ± 1 .42 ,emotional score 1 .50 ± 1 .70 ,physical score 1 .45 ± 1 .46 ,total composite score 4 .05 ± 3 .66) ,and the difference was statistically significant (P<0 .05) .There was no statistically significant difference comparing to the control group (P>0 .05) .Conclusion CRP is effective to treat BPPV .The DHI is available for the evaluation of QOL of BPPV patients .

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