[Comparison of single infusion of anti-BCMA versus combined infusion of anti-CD19 chimeric antigen receptor T cells for immune reconstruction in relapsed/refractory multiple myeloma].

Objective: We observed and compared the differences in immune reconstruction between single-infusion anti-B-cell maturation antigen (BCMA) , chimeric antigen receptor T cells (CAR-T) , and combined infusion of anti-CD19 CAR-T cells in the treatment of recurrent/refractory multiple myeloma (RRMM) . Methods: Sixty-one patients with RRMM who underwent CAR-T cell therapy in our hospital from June 2017 to December 2020 were selected. Among them, 26 patients received anti-BCMA target, and 35 patients received anti-BCMA combined with anti-CD19 target. Using flow cytometry, we determined T cell subsets (CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+)) , B cells (CD19(+)) , and NK cells (CD16(+) CD56(+)) at different time points before and after CAR-T treatment, and detected immunoglobulin IgG, IgA and IgM levels by immunoturbidimetry. We compared the reconstruction rules of lymphocyte subsets and immunoglobulins in the two groups. Results: CD8(+) T lymphocytes recovered most rapidly after the infusion of CAR-T cells, returning to pre-infusion levels at 3 months and 1 month after infusion, respectively[BCMA: 695 (357, 1264) /µl vs 424 (280, 646) /µl; BCMA+CD19: 546 (279, 1672) /µl vs 314 (214, 466) /µl]. NK cells returned to normal levels at 3 months after infusion in both groups[BCMA: 171 (120, 244) /µl, BCMA+CD19: 153 (101, 218) /µl (Normal reference range 150-1100/µl) ]; however, the NK cells were not maintained at stable levels in the BCMA CAR-T cells group. The recovery of CD4(+) T lymphocytes in both groups was slow and remained persistently low within 12 months after infusion, and no recovery was observed in most patients. The reversal of the ratio of CD4(+)/CD8(+) lasted for more than a year. The levels of CD19(+) B cells in both groups returned to baseline 3 months after infusion[BCMA: 62 (10, 72) /µl vs 57 (24, 78) /µl; BCMA+CD19: 40 (4, 94) /µl vs 29 (14, 46) /µl]. IgG returned to the pre-infusion level 12 months after infusion in the group with anti-BCMA cells alone, but not in the group with combined infusion of CD19 CAR T cells[7.82 (6.03, 9.64) g/L vs 6.92 (4.62, 12.76) g/L]. IgA returned to pre-infusion levels at 9 and 12 months after infusion, respectively[BCMA: 0.46 (0.07, 0.51) g/L vs 0.22 (0.12, 4.01) g/L; BCMA+CD19: 0.46 (0.22, 0.98) g/L vs 0.27 (0.10, 0.53) g/L]. IgM in both groups returned to pre-infusion levels 6 months after infusion[BCMA: 0.43 (0.06, 0.60) g/L vs 0.20 (0.13, 0.37) g/L; BCMA+CD19: 0.53 (0.10, 0.80) g/L vs 0.16 (0.11, 0.28) g/L]. There was no significant difference in the indexes of lymphocyte subpopulation reconstruction and immunoglobulin recovery between the two groups at each time point. Conclusion: This study showed that in patients with RRMM treated with CAR-T cells, the appropriate target antigen can be selected without considering the difference of immune reconstruction between anti-BCMA CAR-T and combined anti-CD19 CAR-T therapy.

[Active screening of intestinal carbapenem-resistant Enterobacteriaceae in high-risk patients admitted to the hematology wards and its effect evaluation].

Objective: To evaluate the effect of intestinal carbapenem-resistant Enterobacteriaceae (CRE) active screening combined with enhanced intervention in the prevention and control of nosocomial infection in patients admitted to the hematological ward. Methods: Patients who were admitted to the Department of Hematology in a tertiary-care general hospital from March 1, 2017 to December 31, 2019 and underwent chemotherapy or immunosuppressive therapy comprised the intervention group. They were screened for intestinal CRE at least thrice. From December 1, 2016 to February 28, 2017, patients who underwent chemotherapy or immunosuppressive therapy without active intestinal CRE screening in the Department of Hematology formed the control group. Both the patient groups were monitored for CRE infection in real time. The χ(2) test was used to compare the changes in the CRE infection rate and mortality in high-risk patients before and after the active screening. Results: During the intervention period, the CRE colonization rate of patients was 16.46% (66/401) ; in terms of disease distribution, the colonization rate of acute leukemia was the highest 23.03% (26/113) . Of the 66 colonized patients, 27 (40.9%) patients were identified as positive for CRE at the first screening, 15 (22.7%) were identified at the time of the second screening, and the remaining 24 (36.4%) were identified at the third or subsequent screening; Carbapenem-resistant Klebsiella pneumoniae (CRPK) strains were dominant among the pathogens, accounting for 54.55% (36/66) . During the active screening period, the CRE infection rate (2.49%) and mortality rate (50.00%) of high-risk patients were significantly lower than those of the controls (11.30% and 69.23%, respectively) . The pathogens of 10 CRE infection patients during the intervention period were exactly the same as the previous active screening pathogens, and the coincidence rate was 100%. Conclusion: The CRE colonization rate was the highest in patients with acute leukemia who were admitted in the hematology wards. CRPK is the main pathogen of CRE colonization, infection, and death. Increasing the frequency of screening can significantly raise the positive rate of screening, Active screening can effectively reduce the incidence and subsequent mortality of CRE in high-risk patients admitted in the hematological wards. High coincidence rate between CRE screening positive pathogens and subsequent CRE infection pathogens. Intestinal CRE screening can serve as an indicator of CRE bloodstream infection in patients with hematological diseases as well as provide information for antibiotics therapy.

Effects of a nutrient-dense formula compared with a post-discharge formula on post-discharge growth of preterm very low birth weight infants with extrauterine growth retardation: a multicentre randomised study in China.

BACKGROUND: Post-discharge optimal growth and nutritional intake have beneficial effects for neurodevelopment in preterm very low birth weight infants (VLBWIs) with extrauterine growth retardation (EUGR). The present study aimed to compare the effects of a nutrient-dense formula (NDF) to a post-discharge formula (PDF) on post-discharge growth of preterm VLBWIs with EUGR. METHODS: Forty-eight preterm VLBWIs with EUGR at discharge were randomised to receive NDF (100 kcal per 100 mL; 2.6 g protein per 100 mL) or PDF (74 kcal per 100 mL; 1.95 g protein per 100 mL) for 1-6 months until body weight reached the 50th percentile on growth charts with corrected age. Volume, nutrient intake, anthropometry and biochemistry data were collected. RESULTS: Volume intake was lower in the NDF group than the PDF group during the first 2 months of feeding (P = 0.039 and 0.018, respectively).There were no significant differences in volume intake during months 2-6 of feeding. Energy, protein, carbohydrate and fat intake were higher in the NDF group during months 1-6 of feeding. There were no significant differences in weight, length, and head circumference Z-scores during months 1-6 between the two groups. The △length Z-score from discharge to month 6 was significantly higher in the NDF group than the PDF group (P = 0.043). No differences existed between the two groups with respect to biochemistry. CONCLUSIONS: After discharge, preterm VLBWIs with EUGR fed a NDF gain anthropometric parameter Z-scores similar to those for a PDF within 6 months of follow-up. A NDF leading to gain in length requires further follow-up.

[Expression characteristics and prognosis significance of miRNA-181a in acute myeloid leukemia with normal karyotype].

Objective: To study the expression of miRNA-181a in acute myeloid leukemia (AML) patients with normal karyotype to probe its prognosis significance. Methods: The expression level of miRNA-181a in bone marrow mononuclear cells of 120 de novo AML patients with normal karyotype was detected by real time fluorescence quantitative PCR. The direct sequencing method was used to detect IDH1, IDH2, NPM1, FLT3-ITD, DNMT3A and CEBPα mutations in CN-AML patients after PCR. The relationship between miRNA-181a expression and gene mutation, the clinical parameters and prognosis were analyzed. Results: The rates of overall surviva1 (OS) in high expression and low expression groups were 25.0 months and 15.0 months, respectively (P<0.05) . Relapse free survival (RFS) in high expression and low expression groups were 21.4 months and 11.2 months, respectively (P<0.05) . Significantly higher level hemoglobin, complete remission rate and proportion of wild type NPM1 expression in the high expression of miRNA-181a group were observed when compared with the lower expression of miRNA-181a group (P<0.05) . Multivariate Cox regression analysis showed miRNA-181a overexpression was an independent prognostic factor for CN-AML (HR=2.219, 95%CI 1.601~2.432, P=0.018) . Conclusion: Higher expression of miRNA-181a was a good prognostic factor independent of clinical parameters and high frequency gene mutations, which implicated that the miRNA-181a expression level could be used as an important prognostic indicator of AML patients with normal karyotype.

Oxidative stress inhibits growth and induces apoptotic cell death in human U251 glioma cells via the caspase-3-dependent pathway.

OBJECTIVE: To investigate the possible pathway involved in hydrogen peroxide (H2O2) induced apoptosis in U251 glioma cells. MATERIALS AND METHODS: The cultured U251 glioma cells were used in this study. The cells were divided into three groups: control group (untreated glioma cells), H2O2 group (treated with 100, 300 and 500 µM H2O2) and CI group (treated with 300 µM H2O2 and 15 µM caspase inhibitor, CI). The cellular viability was determined by MTT [3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide] assay. A flow cytometer was used for measuring the cell cycles. The mode of cell death was assessed by Annexin-V/PI-flow cytometry analysis. Fluorescence dihydroethidium (DHE) method was conducted to detect the oxygen species (O2-). Western blot analysis was performed to confirm the pro-caspase-3, caspase-3 and PARP (poly-ADP-ribose polymerase) protein expression. RESULTS: The oxidative stress to U251 glioma cells exhibited in a dose-dependent manner with H2O2 concentrations increasing. The cell viability was considerably decreased and apoptosis occurred in H2O2 treated cells. A G1 cell cycle arrest and O2- level increase were found in H2O2 group. Western blot analysis showed a decrease of pro-caspase-3 protein level and an increase of caspase-3 and PARP level in 300 µM H2O2 treated cells. The H2O2 induced apoptosis depicted above was significantly restrained by CI. CONCLUSIONS: Oxidative stress inhibits growth and induces apoptotic cell death in human U251 glioma cells via the caspase-3-dependent pathway. Mitochondrial pathway might involve in this signaling conduction. These findings are favorable for understanding the mechanisms of oxidative stress-induced apoptosis in U251 glioma cells.

Isolation and characterization of Thermus bacteriophages.

One-hundred-fifteen bacteriophage strains were isolated from alkaline hot springs in Iceland, New Zealand, Russia (Kamchatka), and the U.S.A. The phages belonged to the Myoviridae, Siphoviridae, Tectiviridae, and Inoviridae families. Over 50% of isolates were isometric or filamentous. One type of siphovirus had giant tails of over 800 nm in length. Phages were further characterized by host range, genome size, DNA restriction endonuclease digestion patterns, and temperature and pH sensitivity. Myoviruses and tectiviruses had a worldwide distribution. Most phages were narrowly host-specific and all were highly resistant against heating and alkaline and acidic pH. This is the first time that tectiviruses and filamentous phages are reported for bacteria of the Thermus-Deinococcus phylum. The presence of tectiviruses, inoviruses, and myoviruses is attributed to acquisition from ancestral gamma-proteobacteria by horizontal gene transfer.

[Gene clone of autoantigen hnRNP A2/B1 and research on expression in synovium].

OBJECTIVE: To investigate hnRNPA2/B1's role in rheumatoid synovitis by obtaining its cDNA sequence and analyzing its expression in synovium. METHODS: Total RNA was isolated from peripheral blood mononuclear cells (PBMC) and the RT-PCR was performed. The fragment was cloned into PUC-T1 plasmids and further sequenced. We detected it's expression in synovium with monoclonal antibodies and specific cDNA probes by immunohistochemistry and in situ hybridization. RESULTS: The fragment was identified by DNA sequencing. The expression in rheumatoid synovium was higher than that from osteoarthritis and the normal controls (P < 0.05). CONCLUSION: hnRNP A2/B1 cDNA has been cloned successfully and hnRNP A2/B1 might play an important role in rheumatoid synovitis.

ERPs of characters of Chinese words compared with tone and picture stimuli in adolescents and aged persons.

ERPs to target stimulation with tone, picture and characters of Chinese words ([symbol: see text] CCW were target stimulus) were studied in adolescents group and aged persons group. The age of the adolescent group ranged from 15-17 years, with 30 males and 30 females; the age of the elderly persons group ranged from 60-79 years with 29 males and 18 females. The latencies of N2 of aged persons in tone stimulus and in picture stimulus among males were longer than those of adolescents. The latencies of P3 and RT of aged persons were longer than those of adolescents in three stimulus models. The gender differences of latencies of N2 and RT were not obvious in two groups, but were apparent in P3 in adolescents. The latencies of N2, P3 and RT of CCW stimulus were longer compared to those of picture stimulus in the adolescent group, but not in the aged persons group.

Involvement of NMDA receptor mechanisms in jaw electromyographic activity and plasma extravasation induced by inflammatory irritant application to temporomandibular joint region of rats.

The aim of this study was to examine the possible role of N-methyl-D-aspartate (NMDA) receptor mechanisms in responses induced by the small-fibre excitant and inflammatory irritant mustard oil injected into the temporomandibular joint (TMJ) region of rats. The effects of the non-competitive NMDA antagonist MK-801 were tested on the mustard oil-evoked increases in electromyographic (EMG) activity of the masseter and digastric muscles and Evans Blue plasma extravasation. Five minutes before the mustard oil injection, MK-801 or its vehicle was administered systemically (i.v.), into the third ventricle (i.c.v.), or locally into the TMJ region. Compared with control animals receiving vehicle, the rats receiving MK-801 at an i.v. dose of 0.5 mg/kg (n = 5) showed a significant reduction in the incidence and magnitude of EMG responses as well as in the plasma extravasation evoked by mustard oil; MK-801 at an i.v. dose of 0.1 mg/kg (n = 5) had no significant effect on plasma extravasation or on the incidence and magnitude of EMG responses but did significantly increase the latency of EMG responses. An i.c.v. dose of 0.1 mg/kg (n = 5) or 0.01 mg/kg (n = 5) had no significant effect on plasma extravasation or incidence of EMG responses but did significantly reduce the magnitudes of the masseter EMG response; the 0.01 mg/kg dose also significantly increased the latency of the digastric EMG response. The magnitudes of both the masseter and digastric EMG responses were also significantly reduced by MK-801 administered into the TMJ region at a dose of 0.1 mg/kg (n = 5) but not by 0.01 mg/kg (n = 5); neither dose significantly affected the incidence of EMG responses or the plasma extravasation. These data suggest that both central and peripheral NMDA receptor mechanisms may play an important role in EMG responses evoked by the small-fibre excitant and inflammatory irritant mustard oil, but that different neurochemical mechanisms may be involved in the plasma extravasation induced by mustard oil.

Phage typing of Bacillus subtilis and B. thuringiensis.

Phage typing schemes for Bacillus subtilis and B. thuringiensis were constructed using 98 phages and 743 bacterial strains. Most phages were host-species-specific. Phages were classified by electron microscopy. The B. subtilis scheme includes 10 phages and 29 phage types. The B. thuringiensis scheme comprises 8 phages and 25 phage types and can be applied to B. cereus. There is no correlation between H antigen serotypes and phagovars in B. thuringiensis. Characteristics of typing phages are described for identity control.

[The abnormalities of the cellular immunity in rheumatoid synovium].

With the technique of immunohistochemical (ABC) method monoclonal antibodies were used to identify the lymphocyte subsets, macrophage and the expression of class II MHC (HLA-DR, HLA-DQ) antigens in the synovium cells from 18 RA patients and 8 patients with osteoarthritis as control. The results showed that the main cellular abnormality at the sublayer of the synovium was the appearance of lymphoid follicles which mainly consisted of the infiltration of T lymphocytes (77.8%).50.6% of them were CD4+, which mainly consisted by of CD45RO+ cells in the rheumatoid synovium on the consecutive sections. The increased ratio of CD4/CD8 in RA patients was significantly higher when compared with that in the controls (2.11 +/- 0.93 vs 0.63 +/- 0.13, P < 0.001). In advanced RA with fibrosis of joints, the ratio tended to decrease and was accompanied with reduction of infiltrated lymphocytes. Compared with T cells CD20+ B lymphocyte not only had a lower percentage (25.2%), but also showed a characteristic picture of locating in the centre of the follicles. The fact that most of the CD4+ T cells was helper memorized lymphocytes with CD4 phenotype of positive TAC(+) (interleukin-2 receptor) and that up to 54.2% was anti-HLA-DR and 54.1% anti-HLA-DQ monoclonal antibodies indicated that these T lymphocytes were activated in vivo. Cells with anti-CD68+ were seen all over the RA synovium. Class II HLA and CD68 molecule were also expressed on the endothelium cells of the small vessels. It is suggested that the activated lymphocytes, macrophages and endothelium cells and their abnormal distribution may indicate the abnormalities of the cellular immunity in rheumatoid synovium.

New Bacillus bacteriophage species.

Nine new species of tailed Bacillus phages, based on morphological and physicochemical properties, are defined. Phage P10 is one of the largest viruses known. The total number of tailed Bacillus phage species is presently 33.

The 1958 and 1987 ARA revised criteria for rheumatoid arthritis in Chinese patients. A comparative study.

The American Rheumatism Association (ARA) 1958 and 1987 criteria for the classification of rheumatoid arthritis (RA) were applied to 111 consecutive RA patients and 54 patients with non-RA connective tissue diseases from three hospitals of tertiary level in Beijing. Comparison of the two criteria showed that the specificity was the same, being 88% for both, whereas the sensitivity varied from 94% of the 1958 criteria to 91% of the 1987 criteria. Factors affecting the sensitivity were morning stiffness for more than one hour and increase of number of swollen joints from one to three in the 1987 revised criteria.

[Analysis of knee arthroscopic findings in rheumatic diseases].

46 patients with knee joint arthritis were subjected to arthroscopic examination. The results were analyzed and its clinical significance was discussed. The postoperative diagnoses were rheumatoid arthritis in 19 cases, Sjögren syndrome in one, seronegative spondyloarthropathy in 4, gouty arthritis in one, tuberculous arthritis in one, pigmented villous nodular synovitis in two, undefined synovitis in 5, osteoarthritis in 8 and nonsynovitis conditions in 5. The most important indication of arthroscopic examination is for differential diagnosis of arthritis of unknown cause, such as rheumatoid arthritis with single knee joint involvement, osteoarthritis with symmetric large and small joints involvement as well as crystal induced arthritis. These conditions showed under arthroscope their special features in synovium both grossly, microscopically and immuno-pathologically. The synovial damage as observed through arthroscopic examination correlated to certain extent with the X-ray film changes of the knee joint, but direct visualization with the former is better than indirect imaging with the latter.

[Properties of two Bacillus subtilis bacteriophages].

Two lytic bacteriophages of Bacillus subtilis BF7658 were isolated. Their morphology, biological properties and physiochemical characteristics of their DNAs were compared. Electromicroscopic observation indicated that BS31 has a hexagonal head and contracted tail sheath, BS32 is a complex short-tailed phage and similar to phi 29 but differs from phi 29 in size and other properties. Two phages have a narrow spectrum of host range. The molecular weight of BS31 and BS32 DNAs are 62kb and 17kb respectively by EcoR1 endonuclease analysis. The G + C content of the DNAs are 45.7%(BS31) and 40.7%(BS32). Structural proteins of BS31 gave two major bands and at least ten minor ones; BS32 gave three major bands and six minor ones by SDS-polyacralamide gel electrophoresis.

[The clinical significance of synovial fluid examination].

Synovial fluids from patients were examined, including 15 cases of rheumatoid arthritis (RA); 9 cases of sero-negative arthropathy (3 cases of ankylosing spondylitis, 6 cases of reactive arthritis); 1 case of gout arthritis; and 5 cases of osteoarthritis. The synovial fluids contained 330-72600 white blood cells/mm3, 9-97% of which were granulocytes, in RA patients. Other physical features of RA fluids were: straw green color, cloudy or unclear; moderate viscosity; fair or poor mucin clotting; lack of crystals and bacteria. Synthesis of the above may contribute to the diagnosis of RA. We also found that IgG, IgA and IgM of serum and IgG of synovial fluids in 15 cases of RA were higher than in the other arthritis groups; the IgM-RF positive rates of RA synovial fluids and sera were 20% and 35.7% respectively, while all were negative in the other groups. Immunoglobulins and complement levels in the synovial fluids of all the tested arthritis cases were about half of those in the serum.