CD19-targeted chimeric antigen receptor T-cell (CAR-T) therapy is effective in refractory/relapsed (R/R) B-cell acute lymphoblastic leukemia (B-ALL).This review focuses on achievements,current obstacles,and future directions in CAR-T research.A high complete remission rate of 68 ％ to 93％ could be achieved after and-CD19 CAR-T treatment for B-ALL Cytokine release syndrome and CAR-T-related neurotoxicity could be managed.In view of difficulties collecting autologous lymphocytes,universal CAR-T is a direction to explore.Regarding the high relapse rate after anti-CD19 CAR-T therapy,the main solutions have been developing new targets including CD22 CAR-T,or CD19/CD22 dual CAR-T.Additionally,some studies showed that bridging into transplant postCAR-T could improve leukemia-free survival.Some patients who did not respond to CAR-T therapy were found to have an abnormal conformation of the CD 19 exon or trogocytosis.Anti-CD19 CAR-T therapy for R/R B-ALL is effective.From individual to universal CAR-T,from one target to multi-targets,CAR-T-cell has a chance to be off the shelf in the future.

Background Stromal-derived factor 1α /chemokine receptor 4(SDF-1α/CXCR4) axis is one of the important signals which mediates several different activities such as chemotaxis,adhesion,proliferation and survival resulting in recruitment to sites of immune and inflammatory reactions.Considerable evidence suggests that CXCR4/SDF-1α axis is involved in tumor angiogenesis and plays a key role in the development of ocular neovascularization.Objective The purpose of this study was to explore the effect of CXCR4 antagonist on the development of cxperimental corneal neovascularization(CNV).Methods CNV model was established in the left eye of 8-weekold clean BALB/c mouse by putting the filter with 1 mol/L NaOH at the central cornea for 40 seconds.The animals were randomizcd into hyaluronate group and CXCR4 antagonist group,and the edydrops was topically administered respectively on the day of modeling 4 times per day for 14 days.CNV was examined under the slit lamp at the fourteenth day,and then the corneal suspension and section were made.Expressions of CXCR4 mRNA and protein in corneas were detected using RT-PCR and Western blot.The CD31 level in cornea was assayed by flowcytometry and immunochemistry.The expression of VEGF in burned corneas and suspension from mouse peritoneal macrophages stimulated with CXCR4 antagonist in vitro was detected by ELISA.The use of the animal followed Ragulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Two weeks after corneal alkali burn,the growth of CNV peaked under the slit lamp.Compared with hyaluronate group,CNV was obviously decreased in the CXCR4 antagonist group.Immunochemistry showed that intensity of positive staining for CD31 in cornea in the CXCR4 antagonist group was weaker than the hyaluronate group.Flowcytometry clarified that CD31 positive cells rate was 9.50％ ±2.34％ in the CXCR4 antagonist group and 17.50％ ±3.16％ in the hyaluronate group,showing a significant difference between them (t=-7.312,P＜0.05).In 2,4,7 days after cornea alkali burn,the expressions of CXCR4 mRNA and protein were significantly enhanced in burn corneas compared with normal corneas(P＜0.01 ;P＜0.05).ELISA showed that the VEGF expression level in corneal tissue and supernatant of mouse peritoneal macrophages in vitro were significantly lower in the CXCR4 antagonist group than that of hyaluronate group(t =10.927,5.151,P＜0.05).The expression level of VEGF in corneal suspension was lower in the GM-CSH+CXCR4 antogonist group than that in the GM-CSH group (P＜0.05).Conclusions CXCR4 antagonist can reduce experimental CNV by down-regulating VEGF expression in cornea.

Objective To study the molecular mechanism of interleukin 25 (IL-25)expression in the lung of asthmatic rats.Methods The expressions of IL-25 mRNA and protein in the lungs were detected by Real-time PCR and ELISA,respectively.The levels of IL-25 mRNA and protein were detected by ovalbumin (OVA)in human bronchial epithelial cells.And the transcription factors that regulate IL-2 5 expression were explored through site prediction.Results The expressions of IL-25 mRNA and protein in the lung of OVA-induced asthma rats were significantly increased during animal experiments.Cell experiments showed that OVA could increase the expression of IL-2 5 in human bronchial epithelial cells in a dose-dependent manner,and OVA could upregulate the expression of transcription factor AP1.AP1 was found in the promoter region of IL-25 by site prediction.The AP1 inhibitor (T5224)significantly reduced the expression of IL-25 in OVA-induced human bronchial epithelial cells. Conclusion The molecular mechanism of IL-25 expression induced by OVA in asthma is related to the increase of transcription factor AP1 .

Aim To establish a co-incubation system in cardiac fibroblasts of SD neonatal rats and spleen CD4+ CD25 + regulatory T lymphocytes (Tregs) of normal adult SD rats,and to investigate the effects of eplerenone(EPL) on the interaction of two cells and the relationship with the Kvl.3 channel on Tregs cell membrane.Methods The spleen Tregs of normal adult SD rats were sorted by immunomagnetic bead sorting,and the myocardial fibroblasts of SD rats were isolated by differential adherence method.The experiment was conducted in the following groups:CFs,CFs + Tregs,CFs + Tregs + EPL,Tregs.The proliferation of CFs was detected by CCK-8 method.The expression levels of type Ⅰ collagen,type m collagen and matrix metalloproteinase 2 (MMP-2) secreted by CFs were detected by ELISA.The mRNA expression levels of Kv1.3,KCa3.1 on Tregs cell membrane and intracellular CRAC channel were detected by RT-qPCR technique.Tregs cell membrane Kvl.3 channel protein expression levels were determined by In-Cell Western blot.Results After 48 h incubation of the co-culture system,the cell proliferation was stable.CFs proliferation was marked(P ＜0.01),which could be inhibited by EPL(P ＜0.01).The type Ⅰ,type Ⅲ collagen and MMP-2 secreted by CFs increased (P ＜ 0.01).The expression levels of Kv1.3,KCa3.1 and CRAC channel mRNA in Tregs increased by 6.95,1.99 and 1.53 fold (CFs + Tregs vs Tregs,P ＜0.01),EPL decreased the mRNA level of each channel (CFs +Tregs + EPLvs CFs + Tregs,P＜0.01),and the decrease of Kv1.3 channel was the most significant (P ＜ 0.01).The Kv1.3 channel protein of Tregs increased by 67.9％ (CFs + Tregs vs Tregs,P ＜0.01),which could be inhibited by EPL(P ＜ 0.01).Conclusions Tregs cultured with CFs after 48 h can significantly promote the proliferation of CFs,and EPL can down-regulate the Kv1.3 channel expression on the Tregs membrane and inhibit the activation/proliferation of Tregs,indirectly inhibiting myocardial fibrosis.

The study was aimed to establish a new method of preparation of human placenta factor (PF) and to determine its physic-chemical properties, as well as effects on lymphocytes in vitro. PF was prepared by ultrafiltration. The contents and molecular weight of all constitutions were determined by Bradford method and SDS-PAGE, respectively. Cyclosporin A (CsA) was served as positive control, normal saline (NS) was used as negative control. PHA-stimulated lymphocyte proliferation and mixed lymphocyte reaction (MLR) were detected with MTT assay. The expression of CD69 on T cells was analyzed by flow cytometry. Cytotoxicity of natural killer (NK) cells against K562 tumor cells was examined with LDH release assay. The results indicated that PF was determined to be a group of low molecular weight polypeptides, consisting of two major components whose molecular weight were 9.187 and 4.794 kD respectively. The contents of PF were 5.7 - 6.9 mg/g fresh placenta. PF had similar suppressive effects on PHA-stimulated lymphocyte proliferation and MLR in vitro as compared with CsA (P > 0.05). Both PF and CsA could downregulate the expression of CD69 on T cells which had been stimulated by PMA plus ionomycin (PF vs CsA, P > 0.05). The cytotoxicity of NK cells against K562 cells in PF group was slightly higher or equivalent as compared with that in NS group (P > 0.05), but the cytotoxicity in CsA group was much lower than that in NS group (P < 0.05). It is concluded that a new method of preparation of PF has been established. This study first demonstrates that PF has strong immunosuppressive effects on T cell in vitro, and suppresses T cell proliferation and activation induced by mitogen and alloantigen. This study indicats that PF has no any inhibitory effects, but even enhances the cytotoxicity of NK cells against K562 tumor cells. These results suggest that PF may have suppressive effects on graft-versus-host disease (GVHD) without diminishing graft-versus-tumor (GVT) effects. Therefore, PF may probably be an ideal and promising agent against GVHD.
Cyclosporine ; pharmacology ; Ferritins ; immunology ; isolation & purification ; Graft vs Host Disease ; metabolism ; prevention & control ; Humans ; Immunosuppressive Agents ; immunology ; K562 Cells ; Killer Cells, Natural ; immunology ; Lymphocytes ; immunology ; T-Lymphocytes ; immunology

Modern drying technology was used to explore suitable drying process to provide scientific basis for improving drying processing methods of Scrophulariae Radix. Controlled temperature and humidity drying, vacuum drying apparatus, microwave vacuum drying apparatus, short infrared drying device were used to gain samples for analyzing. The character appearance, concentration of main components and power consumption indicators were chosen for preliminary judging. Six major components, including iridoids and phenylpropanoids were analyzed by UPLC-MS/MS method. The contents of polysaccharides were determined by UV-visible spectrophotometry. The character appearance with controlled temperature and humidity drying and short infrared drying meet the pharmacopoeia standard (Ch. p, edition 2015), while samples with vacuum and microwave vacuum drying apparatus didn't. Compared to fresh sample, concentrations of harpagide, harpagoside, aucubin and catalpol were lower in the dried samples. Angoroside-C showed no significant change before and after drying. Concentration of acteoside increased after drying. Samples with controlled temperature (70 degrees C) and humidity (15% - 10%) drying had high content and short drying time. The better drying process of Scrophulariae Radix was controlled temperature and humidity drying. The method will provide the reference for the drying technology standard of roots medicine.
Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Plant Roots ; chemistry ; Quality Control ; Scrophularia ; chemistry

Objective To investigate the changes in the threshold of auditory brainstem response (ABR),[Ca2+]i and calmodulin(CaM)in cochlear nucleus of the newborn mice infected by murine cytomegalovirus(MCMV)in the brain.Methods Sixty-nine newborn mice were randomized into the model group and the control group.The model group(54 mice)was established by intracranial injection with MCMV viral suspension 20 μl,and the same volume of 0.9％ sodium chloride was injected in the control group(15 mice).After one month,the ABR was tested in a sound-electric screen environment and the threshold was recorded.Then intracellular free calcium [Ca2 +] i and the mRNA level of CaM in the cochlear nucleus were assayed by flow cytometry and RT-PCR.Results Compared to the control group [(64.0 ±1.3)dBSPL],the threshold of ABR in the model group [(84.5 ±2.7)dBSPL] was increased(F =2.789,P =0.000).Moreover,in the model group the intracellular free calcium [Ca2 +] i and the mRNA level of CaM in the cochlear nucleus were increased(F =1.290,P =0.000;F =4.252,P =0.023),the differences were statistically significant.Conclusions The intracranial injection of MCMV can lead to abnormal changes in the threshold of ABR in mice,and the change of [Ca2 +]i/CaM in cochlear nucleus may be the important pathological basis of sensorineural hearing loss induced by MCMV infection.

Bile Ducts, Intrahepatic ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Liver ; metabolism

OBJECTIVETo report a case of pure erythroid leukemia.

METHODSThe clinical features, treatment and prognosis of a rare case of pure erythroid leukemia were reported, and the related literature was reviewed.

RESULTSThe pure erythroid leukemia patient was diagnosed by 90.4% pronormoblasts in bone marrow, 99.5% for erythroid antigen CD71, 67.4% for glycophorin A were detected, while no differentiation antigen of myeloid, lymphoid and megakaryocyte lineages were observed. HAG (homoharringtonine + Cytarabine and G-CSF) regimen were administered with no effect. The patient developed multiple organ failure and died soon.

CONCLUSIONPure erythroid leukemia has a fulminant clinical course with poor response to chemotherapy and worse prognosis.

Humans ; Leukemia, Erythroblastic, Acute ; therapy ; Male ; Middle Aged ; Prognosis

OBJECTIVETo compare therapeutic effects of local injection with autoallergic platelet rich plasma (PRP) or autoallergic whole blood (AWB) for the treatment of chronic tennis elbow.

METHODSFrom January 2011 to January 2014, 40 patients with chronic tennis elbow were divided into 2 groups, 20 cases in each group: PRP group and AWB group. There were 20 patients in PRP group treated with local injection of autoallergic platelet rich plasma, including 5 males and 15 females, with an average age of (47.50 ± 9.86) years old; and the average course of disease was (4.67 ± 3.27) months. Among the 20 patients in AWB group treated with local injection of autoallergic whole blood, 3 patients were male and 17 patients were female, with an average age of (46.50 ± 9.96) years old;and the average course of disease was (4.53 ± 2.27) months. The elbow joint was fixed with elastic stockings after injection. All the patients were guided to do strengthening and extension exercises during the follow-up period. Visual analog scale (VAS), Mayo scores for elbow and pressure pain threshold (PPT) were used to evaluate clinical effects after injection immediately and 4,8 weeks after treatment. Results:All the patients were followed up,there were no infections and swelling occurred. The VAS, Mayo and PPT scores of patients in PRP group were improved from pre-therapy 7.22 ± 1.32, 56.71 ± 10.90 and 17.47 ± 4.62 to 2.73 ± 1.00, 91.59 ± 6.95 and 21.35 ± 4.80 respectively 8 weeks after treatment. The VAS, Mayo and PPT scores of patients in AWB group were improved from pre-therapy 7.16 ±1.27, 54.72 ± 8.36 and 17.06 ± 4.83 to 3.81 ± 1.36, 82.06 ± 7.89 and 20.12 ± 4.97 respectively 8 weeks after treatment. All the pain and functional variables including VAS, PPT, and Mayo scores were improved significantly in both groups 4 weeks after injection. On the 4th week after injection, there was no statistically significant difference in PPT between two groups; while the VAS and Mayo score of AWB group were lower than those of PRP group. On the 8th week after injection, the VAS of AWB group was higher than that of PRP group; but the Mayo and PPT scores of AWB group were lower than those of PRP group.

CONCLUSIONPRP and AWB injections are both effective to treat chronic lateral epicondylitis. Compared with AWB injection, PRP injection may be more effective in releasing pain and improving function for a longer time.

Adult ; Blood Transfusion ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Pain Threshold ; Platelet-Rich Plasma ; Tennis Elbow ; therapy ; Visual Analog Scale