Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.

Progress of chimeric antigen receptor T cells in the treatment of hematologic malignancies：reports from the 57th American Society of Hematology annual meeting / 白血病·淋巴瘤

Chimeric antigen receptor T cells (CAR-T), one of the most promising cancer immunotherapy, has attracted much attention in the 57th American Society of Hematology (ASH) annual meeting. CAR-T therapy has obtained significant effect on leukemia and lymphoma and the latest research results are also inspiring in the 57th ASH annual meeting. It is an important task that how to combine CAR-T therapy with the traditional methods of treatment and the immune checkpoint blocking antibodies and small-molecule-targeted drugs to achieve the best effect. This paper will review the progress of CAR-T in the treatment of hematologic malignancies.

Progress of targeted therapy in diffuse large B-cell lymphoma: reports in the 55th ASH annual meeting / 白血病·淋巴瘤

With the further study on heterogeneity and molecular mechanism of diffuse large B-cell lymphoma (DLBCL),many novel targeted drugs have been developed,which also showed clinical benefit in early-stage studies.Therapeutic targets for DLBCL mainly include the surface antigens of B lymphocyte,B cell receptor signaling and the cellular microenvironment.Several new clinical trials about targeted therapy for DLBCL had been reported in the 55th ASH annual meeting,and the results were encouraging.The advances in this field will be summarized in this paper based on the new reports in the 55th ASH annual meeting.

The integrin linked kinase promotes migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB pathway / 中国综合临床

Objective To investigate the role of integrin-linked kinase (ILK) on migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB (NF-κB) pathway.Methods A549 cell line were overexpressed ILK and matrix metalloproteinase-9 (MMP-9) confirmed by cell transfection,siRNA interference,cell scratch test,real-time quantitative PCR and Western Blot.Results Over-expression of ILK stimulated MMP-9 expression in lung cancer cells(P ＜ 0.01).The addition of MMP-9 inhibitor doxycycline and anti-MMP-9 neutralizing antibody significantly impaired the wound healing capacity of ILK-transfected cells(P ＜ 0.01),as well as by in vitro matrigel invasion assay (P ＜ 0.01).In addition,overexpression ILK induced phosphorylation and nuclear translocation of nuclear factor-κB subunit p65.Upregulation of MMP-9 was severely abolished by either BAY 11-7028,a specific NF-κB inhibitor,or siRNA targeted to NF-κB p65 in ILK over-expression cells.Conclusion The finding indicate that over-expression of ILK can promote the migration and invasion of lung cancer cell,and upregulate MMP-9 through the NF-κB pathway.

High risk factors in 128 elderly patients undergoing hip replacement / 中国组织工程研究

BACKGROUND:Deep vein thrombosis after hip replacement has a high incidence rate. Moreover, deep vein thrombosis can induce pulmonary embolism that can endanger patients’ life and dysfunction of distant deep vein. The appearance of deep vein thrombosis is a great obstacle for the gradual y increased hip replacement. OBJECTIVE:To observe the occurrence of deep vein thrombosis of lower limb after total hip replacement in elderly patients, and to screen the risk factors for the occurrence of deep vein thrombosis of lower limbs. METHODS:Clinical data of 128 elderly patients with hip replacement were analyzed retrospectively. Al patients were examined with color Doppler ultrasound in double lower limbs at 1 day before replacement and 7 days after replacement. Multifactor unconditional logistic analysis was conducted on clinical related factors and the formation of lower limb deep vein thrombosis. RESULTS AND CONCLUSION:At 7 days after the operation, 16 patients affected deep vein thrombosis of lower limb. The factors for deep vein thrombosis contained female, general anesthesia, bilateral hip replacement and the application of bone cement (P<0.05). The risk for deep vein thrombosis after total hip replacement significantly increased in elderly patients aged over 70 years. Multifactor unconditional logistic analysis exhibited that the multiple risks of sex, obesity and the use of bone cement in elderly patients with deep vein thrombosis increased to 11.398, 3.109 and 8.925. The patients with a blood type O at the age of over 70 years experienced a decreased risk for deep vein thrombosis after total hip replacement. The occurrence of deep vein thrombosis decreased to 0.186 times after replacement. Blood type O could be considered as a protective factor for the occurrence of deep vein thrombosis.

Value of CAT and mMRC in clinical evaluation of chronic obstructive pulmonary disease / 重庆医学

Objective To evaluate the health status ,dyspnea and exercise endurance in the patients with chronic obstructive pul-monary disease(COPD) by the CAT scale ,modified medical research council(mMRC) dyspnea scale and 6-min walk test(6MWT) and to analyze the correlation between their evaluation results with the percentage (FEV1% pred) of the forced expiratory volume at 1 second(FEV1) in the predict value and whether complicating pulmonary hypertension (PH) .Methods 70 patients with COPD were performed the examinations of the lung function ,CAT scale ,mMRC score and 6MWT .The correlation among the various e-valuation systems and between each evaluation system with the severity of the pulmonary function airway limitation were compre -hensively evaluated .The CAT scores ,mMRC scores and 6 min walking distance(6MWD) were compared between the COPD com-plicating PH group and the simple COPD group .Results The CAT scale was negatively correlated with 6MWD(r= -0 .623 ,P0 .05) .The CAT scores had statistical difference between the COPD complicating PH group and the simple COPD group (P<0 .01) .6MWD had the highest accuracy for screening COPD whether complicating PH ,followed by CAT .The diagnostic cut-off point by adopting CAT for judging the risk of COPD complicating PH was 21 .Conclusion The CAT scale has good correlation with 6MWD and no correlation with FEV1% pred .The CAT scale is more suitable for the overall assessment of the total severity of COPD .Patients with the high CAT scores have higher risk of suffering from PH .

Effects of intramuscular and oral mecobalamin in elderly patients with vitamin B12 deficiency / 中国综合临床

Objective To investigate the effect of intramuscular and oral mecobalamin on hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.Methods One hundred and twenty-six patients with vitamin B12 deficiency who fulfilled the inclusion criteria were randomly divided into 3 groups,42 cases in each,including the intramuscular group (to receive mecobalamin therapy intramuscularly),the oral group ( to receive mecobalamin therapy orally) and the control group ( without mecobalamin therapy).The changes of hematologic markers including hemoglobin (Hb,g/L),mean corpuscular volume (MCV,fl),serum levels of vitamin B12 (ng/L),folate and total homocysteine (Hcy,μmol/L),and neurological signs before and after treatment were compared among these groups.Results Baseline characteristics among the three groups were similar.After a 6-month therapy,there were no differences in any markers in control group patients in comparison to baseline;for patients in the intramuscular group,the blood vitamin B12 levels increased from (139.13 ± 31.57)ng/L to (328.10 ± 42.35 )ng/L (P ＜ 0.001 ).Hcy levels decreased from (36.29 ± 16.23 )μmol/L to ( 18.23 ± 9.85 ) μmol/L ( P ＜ 0.001 ).Hb rose from ( 125.34 ± 16.21 ) g/L to ( 132.79 ± 15.98 )g/L (P =0.037).MCV reduced from (92.98 ±5.35)fl to (87.65 ±5.74)fl (P ＜0.001 ) ;For patients in the oral administration group,the blood vitamin B12 levels increased from ( 138.19 ± 29.95) ng/L to (487.79 ±32.21 ) ng/L ( P ＜ 0.001 ).Hcy levels decreased from ( 33.27 ± 11.51 ) μmol/L to ( 17.49 ± 10.13 ) μmol/L( P ＜ 0.001 ).Hb rose from ( 125.89 ± 17.65 ) g/L to ( 133.46 ± 16.26) g/L ( P =0.041 ).MCV reduced from (93.08 ± 5.10 ) fl to ( 89.29 ± 5.37 ) fl ( P =0.001 ).After the 6-month therapy,there were somewhat improvement in MMSE scores of the intramuscular ( 28.24 ± 3.89 vs.27.85 ± 3.56,P =0.633 ) and the oral groups (27.97 ± 3.77 vs.27.34 ± 3.15,P =0.408 ) compared with baseline,but the differences were not significant.The achilles tendon reflex of the intramuscular ( 1.86 ± 0.67 vs.1.56 ± 0.61,P =0.035 ) and the oral groups ( 1.79 ± 0.64 vs.1.43 ± 0.51,P =0.006 ) were enhanced compared with baseline.Foot vibration sensation of the intramuscular ( 1.35 ± 0.37 vs.1.06 ± 0.41,P =0.001 ) and the oral groups ( 1.24 ± 0.52vs.1.01 ± 0.43,P =0.03) were improved compared with baseline.After treatment,the serum VitB12 concentration in the oral group were higher than that of the intramuscular group ( P ＜ 0.001 ).There were no significant differences in other indexes between the two groups.Conclusion Mecobalamin,administered either intramuscularly or orally,may improve the hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.

The clinical effects of long-term domiciliary oxygen therapy and pulmonary rehabilization program on the patients with chronic obstructive pulmonary disease / 中国综合临床

Objective To evaluate the clinical effects of long-term domiciliary oxygen therapy (LDOT)in accompany with pummonary rehabilization program on the patients with chronic obstructive pulmonary disease (COPD).Methods Seventy two COPD cases receiving LDOT treatment were randomized into treatment group and control group.The patients in control group were given LDOT alone,while the treatment group was given pulmonary rehabilization besides LDOT.Lung functions,arterial blood gas parameters and blood rheological parameters were compared between the two groups 2 years after the observation.Results The follow-up period lasted for 1 - 2 years.The frequency of acute exageration in the treatment group ( 3.0 ± 1.3 ) was significantly lower than that of control group (4.0 ± 1.6) ( t =1.893,P ＜ 0.05 ).Compared with that of control group,the FEV1([1.59±0.08]L vs.[1.41 ±0.13]L,t =-3.966,P ＜0.01),FVC ([2.47 ±0.20]L vs.[2.27 ±0.17]L,t=-2.788,P＜0.05),FEV1％ ([2.47±0.20]％ vs.[2.27±0.17]L,t=-4.402,P＜0.01) and PaO2 ( [79.1 ± 8.9 ] kPa vs.[ 60.0 ± 6.6 ] kPa,t =- 4.622,P ＜ 0.01 ) were significantly increased,while plasma viscosity ( [ 2.14 ± 0.31] mPa · s vs.[ 2.44 ± 0.45 ] mPa · s,t =1.985,P ＜ 0.05 ),Low shear blood viscosity ( [ 13.48 ± 1.97 ] mPa · s vs.[ 14.33 ± 1.87 ] mPa · s,t =2.126,P ＜ 0.05 ),median shear whole blood viscosity( [ 6.33 ± 0.66 ] mPa · s vs.(7.92 ± 0.98 ) mPa · s,t =4.238,P ＜ 0.01 ),high shear whole blood viscosity ([4.58 ±0.59] mPa · s vs.[5.33 ±0.68]mPa · s,t =0.3890,P ＜0.01) and erythrocyte sedimentation rate ( [ 30.63 ± 5.76 ] mm/1 h vs.[ 35.63 ± 6.925 ] mm/1 h,t =2.230,P ＜ 0.05 ) was greatly decrease.Conclusion Long-term domiciliary oxygen therapy in company with pulmonary rehabilization program is helpful to improve the lung function,arterial blood gas parameters and rheological status of COPD patients.

Clinical significance of detection of bcr-abl mRNA in chronic myeloid leukemia / 白血病·淋巴瘤

Objective To evaluate significance of the quantification of bcr-abl mRNA in diagnosis and therapy of chronic myeloid leukemia (CML),essentiality significance for monitoring minimal residual disease. Methods Bcr-abl mRNA of 518 CML patients were detected using real-time PCR. Results Expression of bcr-abl mRNA was gradually increased among blastic phase (BP) (12.6 %),accelerated phase (AP) (25.4 %) and chronic phase (CP) (57.2 %) (P<0.05). Quantification of bcr-abl mRNA was cut down gradually after allotransplantation in the patients and becomes normal after treatment for 6 months. But quantification of bcr-abl mRNA inpatients treated with imatinib mesylate became normal after 12 months. Conclusion Real-time PCR was reliable and can be used for diagnosis,monitoring the treatment outcome,detecting the minimal residual disease,and predicting blast crisis.

Effects of heparin and low molecular weight heparin on serum level of pregnancy associated plasma protein A in patients with cerebrovascular disease / 中华检验医学杂志

Objecfive To investigate the effects of treatment for cerebrovascular disorder patients with heparin and low molecular weight heparin(LMWH) on serum PAPP-A concentrations and provide the basis for evaluating the clinical significance of PAPP-A in the following study.Methods Forty cases with cerebrovascular disease from Qilu Hospital from November 2009 to May 2010 were collected in this study.Blood samples were taken before and after drug administration.All cases were divided into four groups according to situation of medication.Group A consisted of 10 patients who received subcutaneous LMWH anticoagulation therapy, and blood samples were collected before LMWH injection, three hours after subcutaneous LMWH anticoagulation therapy in the first day, the second day and the seventh day and 24 hours after the last injection. Group B consisted of 10 patients who did not receive LMWH therapy, and blood samples were collected immediately after admission, the first day, the second day and the seventh day after admission. Group C consisted of 10 patients with percutaneous carotid intervention who received intravenous heparin at the beginning of stenting, and blood samples were collected from the arterial sheath just before angiography and heparin administration, and at 3, 5, 15, 40 and 100 min after heparin administration. Group D consisted of 10 patients who received carotid angiography but LMWH-free therapy,and blood samples were collected from the arterial sheath just before and after angiography. Serum PAPP-A concentrations were analyzed by ELISA to evaluate the differences of intra-groups and differences at different time points of inter-groups. Results In group A, PAPP-A concentrations were time dependent and elevated gradually from 12. 36 (9. 90-14. 32) mIU/L before LMWH injection to 21.80 (23.50-19.73) mIU/L at the seventh day after injection (M=38. 72, P < 0.01 ). In group C, there was a rapid increase of PAPP-A concentration from 12. 86 ( 9. 67-14. 05 ) mIU/L to 51.56 ( 44. 20-66. 00 ) mIU/L within 5 min after intravenous heparin injection (M=46. 06, P <0. 01 ). The PAPP-A concentration of one week after LMWH administration in group A was 21.80 (23.50-19.73) mIU/L, significantly higher than that in group B [11.81 (9. 21-12. 89) mIU/L] (U<0. O01, P<0.01). The PAPP-A concentration at 15 min after heparin administration in group C was 43.70 (37.70-54. 30) mIU/L, significantly higher than that after angiography in group D [14. 18 (11.25-15. 86) mIU/L] ( U<0. 001, P <0. 01 ). The peak level of blood PAPP-A after subcutaneous LMWH injection was significantly lower than that after intravenous heparin injection. The concentrations in group A and C were 21.80 ( 23.50-19. 73 ) and 51.56 (44. 20-66. 00) mIU/L respectively, and had a significant difference ( U=0. 999, P < 0. 01 ) . Conclusions Both intravenous heparin and subcutaneous LMWH administration induce an increase in serum PAPP-A concentration. The effect of drug should be considered when PAPP-A is selected as an evaluation indicator.

Infection in patients with malignant tumors / 中南大学学报(医学版)

OBJECTIVE@#To investigate the characters of infection in patients with malignant tumors, especially the distribution, yearly change of pathogens, and pathogen resistance to common antibacterial agents.@*METHODS@#We respectively analyzed the characters of infection in 489 patients with malignant tumors.@*RESULTS@#The respiratory tract was the most frequent infection site (61.1%). The infection was mainly caused by opportunistic pathogens. The Gram-negative bacterias mainly consisted of Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii (46.3%). The Gram-positive bacteria mainly consisted of Staphylococcus aureus and Staphylococcus epidermidis (29.9%), and the rest 23.8% of the infection was caused by different fungi, mainly consisting of Candida albicans. The ratio of the Gram-negative bacteria resistance to antibiotics such as penicillins, cephalosporins (except ceftazidime), sulfanilamides, tetracyclines and quinolones was higher. The ratio of the Gram-positive bacteria resistance to antibiotics such as penicillins, macrolides and quinolones was higher. The ratio of fungus resistance to antibacterial agents such as fluconazol and itraconazole was higher. The infection caused by fungi obviously increased in the past 5 years.@*CONCLUSION@#The infection in patients with malignant tumors is mainly caused by opportunistic pathogens, and the pathogen resistance to antibacterial agents is serious. The infection caused by fungi is increasing.

Application of hydroxyapatite artificial bone in bilateral open-door posterior cervical expansive laminoplasty / 中国组织工程研究

BACKGROUND:Hydroxyapatite (HA) artificial bone,as bone grafting substitute,would not cause inflammatory reaction or immunological rejection and possesses good biocompatibility after transplantation into human body.It is a novel implant material with bone conduction ability.OBJECTIVE:To investigate the efficacy of HA artificial bone in bilateral open-door posterior cervical expansive laminoplasty and to make a comparison with autogenous bone.DESIGN,TIME AND SETFING:A retrospective case analysis was performed at the Department of Spine Surgery,Hungpuyuan Branch,the First Affiliated Hospital of Sun Yat-sen University from March 2001 to December 2008.PARTICIPANTS:Seventy patients with cervical spondylosis complicated by compression in 3 or more segments or by cervical stenosis and additional fifteen patients with cervical stenosis complicated by cervical trauma were included in this study.METHODS:A bilateral open-door posterior cervical expansive laminoplasty was performed,in which,23 patients received autogenous bone transplantation (autogenous bone group) and 62 patients underwent HA artificial bone transplantation (HA group).MAIN OUTCOME MEASURES:① Japanese Orthopaedic Association (JOA) score pdor to and after surgery,surgery time,and intraoperative bleeding.② HA artificial bone-host biocompatibility.RESULTS:All eighty-five patients were followed up for more than 3 months.There was no significant difference in JOA scores no matter prior to or after surgery between the autogenous bone and HA groups (P＞0.05).The surgery time averaged 85.2 minutes (range 65-110 minutes) in the HA group and averaged 116.4 minutes (range 75-150 minutes) in the autogenous bone group.The intraoperative bleeding averaged 210 mL (range 130-400 mL) in the HA group and averaged 260 mL (range 170-500 mL) in the autogenous bone group.There were no material-host response and other severe complications found in each group,except HA artificial bone fragmentation in 3 patients from the HA group.CONCLUSION:HA artificial bone yields good efficacy and causes fewer complications in bilateral open-door posterior cervical expansive laminoplasty;in addition,it requires less time for surgery and causes less bleeding.

Efficacy of monosegmental pedicle instrumentation in treatment of traumatic thoracolumbar burst fractures / 中华创伤杂志

Objective To evaluate the clinical efficacy of monosegmental pedicle instrumentation in management of thoracolumbar burst fractures. Methods A total of 67 patients with traumatic thora-columbar burst fractures (type A3.1 and A3.2) were treated with monosegmental pedicle instrumentation in our department from October 2003 to February 2008. Imageologic effect was observed by measuring sagittal index and wedge index via X-ray and clinical outcomes evaluated by using low back outcome score. Results All operations were performed successfully, with average operation duration of 93 mi-nutes and average intraoperative blood loss of 157 ml. Of all, 65 patients were followed up for 4-27 months (average 19.8 months), which showed that all the patients achieved bony fusion, with no implant failure except for one with screw loosening. The sagittal index and wedge index were 13.06°and 42.9% preoperatively and 4.47° and 21.78% postoperatively, with statistical difference (P <0.01). The final follow-up showed no significant correction loss except for two patients (P < 0.05). The low back outcome scores of all patients at follow-up were improved significantly (P < 0.05). Conclusions Monoseg-mental pedicle instrumentation has advantages of minimal invasion, short operative duration, less blood loss and less vertebral motion segment loss and hence is an effective and reliable operative technique for thoracolumbar burst fractures.

Epidemiologic analysis of greenhouse farmer's lung in part of rural areas of Liaoning province / 中国实用内科杂志

Objective To survey the prevalence of greenhouse farmer's lung and related risk factors in part of rural areas of Liaoning Province.Methods Using uniform scheme,procedures and questionnaire,a survey for 5420 farmers(2660 men and 2760 women)with complete data who work inside greenhouses was performed in Shenyang,Xinmin,Chaoyang,and Jinzhou between August 2006 and June 2009.Pulmonary function tests was performed for every active farmer.Results Greenhouse farmer's lung was diagnosed in 308 cases,205 men(66.55%,205/308)and 103 women(33.44%,103/308),a prevalence of 5.7%(308/5420).The prevalence rate of greenhouse farmer's lung in males was significantly higher than that in females(?2=39.93,P0.05).In the 308 cases,the number of patiernts presented with fever chill,cough/sputum,chest tightness/shortness of breath were 180(58.44%),192(62.34%),160(51.95%)respectively,and the number of crepitations,radiological changes,spirometry abnormalities and serum IgE antibodies(+)was 164(53.25%),153(49.68%),147(47.73%)and 136(44.16%)at the time of the study.62.34%(192/308)of patients with greenhouse farmer's lung were mild and 38.66%(116/308)were severe.Conclusion The total prevalence rate of greenhouse farmer's lung in part of rural areas of Liaoning Province was 5.7% and multiple risk factors were associated with the disease.

Neonatal Screening for Alpha-thalassemia / 中华围产医学杂志

0.05), but significantly lower than 8.3% in Guangzhou area(P