Objective:To retrospectively analyze efficacy and safety of granulocyte colony-stimulating factor(G-CSF)in relapsed/refractory B cell acute lymphoblastic leukemia(R/R B-ALL)patients with neutropenia(NE)after receiving CAR-T cell therapy.Methods:From March 2017 to December 2022,99 patients with R/R B-ALL developed NE after receiving CAR-T cell therapy in Tianjin First Central Hospital were collected and divided into two groups according to using time of G-CSF.One was early G-CSF group(received G-CSF within 7 days,n=56),the other was control group(received G-CSF after 7 d,n=43),whose recovery of NE and occurrence of adverse reactions after G-CSF were compared.Results:Duration of NE in early G-CSF group was shorter than control group[4(2,5.7)vs 11(9,14),P<0.05],but there were no significant differences in the lowest absolute neutrophil count(ANC),degree of NE inhibition and incidence of infection(P=0.261,P=0.09,P=0.111).There was no significant difference between incidence and severity of cytokine release syndrome(CRS)between two groups,and CRS was controllable in all patients.Conclusion:Early application of G-CSF in R/R B-ALL patients after CAR-T cell therapy can shorten duration of NE,and has no significant effect on adverse reactions after CAR-T cells.

Objective To investigate the clinical value of high resolution vascular wall imaging(HRVWI)in the diagnosis and treatment of central nervous system vasculitis(CNSV)in children.Methods A total of 36 children with CNSV underwent HRVWI examination were selected.The HRVWI imaging features,involved vessel types and stenosis degree were analyzed retrospectively and compared with magnetic resonance angiography(MRA).Combined with some patients'follow-up and reviewed imaging after treatment.Results HRVWI showed that all of the 36 children with CNSV had cerebral arteritis,and the affected inflammatory ves-sels showed varying degrees of focal arterial wall thickening and enhancement,mainly uniform thickening and centripetal enhance-ment of the affected arterial wall.Most were mid-enhanced,and the stenosis degree was mostly at level 1(21/36).According to the type of involvement cerebrovascular,there were 26 cases of large-medium vessel type,6 cases of small vessel type and 4 cases of mixed type.Compared with MRA,HRVWI had a higher sensitivity in diagnosing CNSV(P＜0.05).13 cases were followed up after treat-ment,and HRVWI showed cerebral vasculitis relief in 8 cases and progression in 5 cases,consistent with changes in clinical symp-toms and laboratory indicators.Conclusion The imaging features of CNSV in children with different causes are different.HRVWI has a high sensitivity in the diagnosis of CNSV in children.HRVWI plays an important role in the diagnosis and treatment of CNSV in children.

Objective:To analyze the molecular characteristics of E genes of imported dengue virus (DENV) in Sichuan province from 2012 to 2021, such as serotypes and genotypes, and so to provide the basis for virus traceability.Methods:DENV E genes of the 28 serum samples which were tested DENV positive by real time RT-qPCR from 2012 to 2021 in Sichuan province were amplified and sequenced by RT-PCR. Bioinformatics software Mega Align5.0 was used for the alignment analyses of nucleotide and deduced amino acid sequences, and bioinformatics software Mega7.0 was used to draw phylogenetic trees.Results:RT-PCR and sequencing indicated that 28 E genes of DENV were obtained. The serotypes and genotypes of DENV were revealed by homology and phylogenetic analysis based on E genes of DENV; 20 virus strains of Sichuan belonged to DENV serotype 1 (DENV-1), of which 16 virus strains were genotype I (G-I), 4 virus strains were genotype V (G-V); 7 virus strains belonged to DENV serotype 2 (DENV-2), of which 4 virus strains were G-I, 3 virus strains were genotype II (G-II); 1 virus strain belonged to DENV serotype 3 (DENV-3), G-I. The phylogenetic trees showed that Sichuan strains had more homology with strains of Southeast Asia and Guangdong province in China, that is consistent with the epidemiological data.Conclusions:In 2012 to 2021, the imported DENV in Sichuan province had 3 serotypes and multiple genotypes, and they were most likely transmitted from Southeast Asian countries.

Objective:To explore the diagnostic performance of ultrasound attenuation imaging (ATI) in grading the degree of hepatic steatosis in metabolic dysfunction-associated fatty liver disease (MAFLD).Methods:The liver gray-scale ultrasound and ATI examinations were performed on 212 subjects who were treated in Zhongshan Hospital Affiliated to Fudan University from August 2020 to March 2021. The attenuation coefficient(AC) values among different degrees of hepatic steatosis were analyzed and the diagnostic performance of ATI was evaluated. Relationships between AC values and clinical characteristics were assessed by Pearson′s correlation analysis.Results:The AC values for normal liver, mild, moderate and severe fatty liver were (0.56±0.05)dB·cm -1·MHz -1, (0.68±0.09)dB·cm -1·MHz -1, (0.82±0.09)dB·cm -1·MHz -1, (0.94±0.09)dB·cm -1·MHz -1, respectively. There were significant differences in AC values among different hepatic steatosis divisions( P<0.008). There was highly significant correlation between AC values and the degree of hepatic steatosis( r=0.860, P<0.01), moderate correlation between AC values and BMI( r=0.425, P<0.01), weak correlation between AC values and HDL-C( r=-0.237, P=0.029), no correlations between AC values and age, TC, TG, LDL-C ( r=0.083, 0.055, 0.133, -0.039, all P>0.05) .The areas under the receiver operating characteristics curve of ATI for mild fatty liver and above, moderate fatty liver and above, severe fatty liver and above were 0.958, 0.962, 0.918; the sensitivity were 90.1%, 95.8%, 94.9%, the specificity were 96.1%, 87.1%, 73.9%, and the cut-off values were 0.666 dB·cm -1·MHz -1, 0.719 dB·cm -1·MHz -1, 0.803 dB·cm -1·MHz -1, respectively. Conclusions:ATI is a reliable and convenient method for evaluating the degree of hepatic steatosis in MAFLD.

Objective: To investigate the efficacy and safety of CD19 chimeric antigen receptor T (CAR-T) lymphocytes for the treatment of B cell lymphoma. Methods: A total of 22 patients with B-cell lymphoma from February 1, 2017 to July 1, 2018 were reviewed to evaluate the efficacy and adverse reactions of CD19 CAR-T. Results: Of 22 patients with B-cell lymphoma received CD19 CAR-T cells, the median dose of CAR-T cells was 7.2 (2.0-12.0) ×10⁶/kg. Nine of 12 cases of relapse refractory patients were overall response. Complete remission (CR) occurred in 2 of 12 patients, partial remission (PR) in 7 of 12 patients. The overall response in minor residual disease positive (MRD) group was 8 of 10 patients. CD19 CAR-T cells proliferated in vivo and were detectable in the blood of patients. The peak timepoints of CAR-T cells proliferated in the relapsed refractory and MRD positive groups were 12 (5-19) and 4.5 (1-12) days after treatment respectively, and among peripheral blood cells, CAR-T cells accounted for 10.10% (3.55%-24.74%) and 4.02% (2.23%-28.60%) of T lymphocytes respectively. The MRD positive patients achieved sustained remissions during a median follow-up of 8 months (rang 3-18 months) . None of all the patients relapsed during a median follow-up time of 10 months (3-18 months) . However, 7 PR responders of the relapsed refractory patients maintained a good condition for 1.5-6.0 months. One patient bridged to hematopoietic stem cell transplantation, another one sustained remission for 12 months. Cytokine-release syndrome (CRS) occurred in 14 patients with grade 1-2 CRS in MRD positive group and grade 3 CRS in relapsed refractory group. Conclusions CAR-T cell therapy not only played a role in the rescue treatment of relapsed and refractory patients, but also produced a surprising effect in the consolidation and maintenance of B-cell lymphoma. CD19 CAR-T cells might be more effective in the treatment of MRD positive B-cell lymphoma patients than in the refractory or relapsed cases. High response rate was observed with fewer adverse reactions.

过继细胞疗法（ACT）的飞速发展使其成为肿瘤治疗手段中的一项新热点，其中嵌合抗原受体修饰的T细胞（CAR-T 细胞）在治疗恶性血液肿瘤中取得的成果更是令人振奋，同时也为实体瘤的治疗提供了新策略。但是，目前CAR-T细胞免疫疗法 在肿瘤治疗过程中的局限性也日渐显露。本文旨在针对CAR-T细胞在肿瘤治疗中的研究进展及治疗中的挑战予以简要探讨。

Objective To study the correlation of wake-up stroke with progressive stroke.Methods Three hundred and twelve patients with acute ischemic stroke,admitted to our hospital from January 2014 to December 2015 were divided into progressive stroke group (n=70) and non-progressive stroke group (n=242).Demographic features,clinical characteristics,and incidence of wake-up stroke were compared between the two groups.The association between wake-up stroke and progressive stroke was analyzed.Results The incidence of wake-up stroke,homocysteine level,and fibrinogen level in progressive stroke group were significantly higher than those in the non-progressive stroke group (40.0％ vs.18.2％,P=0.000;[17.486±16.835] μmol/L vs.[14.321±7.251] μmol/L,P=0.023;[3.539±1.009] g/L vs.[3.134±0.775] g/L,P=0.000).Multivariate Logistic regression analysis showed that wake-up stoke,homocysteine and fibrinogen were the independent predictive factors of progressive stroke (OR=2.978,95％CI:1.623-5.464,P=0.000;OR=1.026,95％CI:1.002-1.052,P=0.035;OR=1.800,95％CI:1.310-2.472,P=0.000).Conclusion Wake-up stoke is a predictive factor of progressive stroke.

OBJECTIVETo explore effects of iron overload on hematopoiesis in mice with bone marrow injury and its possible mechanism (s).

METHODSC57BL/6 mice were divided into control, iron, irradiation, irradiation+iron groups. The iron-overloaded model of bone marrow injury was set up after mice were exposed to the dose of 4 Gy total body irradiation and (or) were injected iron dextran intraperitoneally. Iron overload was confirmed by observing iron deposits in mice and bone marrow labile iron pool. Additionally, the number of peripheral blood and bone marrow mononuclear cells and the frequency of erythroid cells and myeloid cells were counted and hematopoietic function was assessed.

RESULTS(1)Iron overload occurred by bone marrow biopsy and flow cytometry analysis. (2)Compared with control group, the number of platelets [(801.9±81.2)×10⁹/L vs (926.0±28.2)×10⁹/L] and BMMNC and the frequency of erythroid cells and myeloid cells decreased. Moreover, hematopoietic colony forming units and single-cell cloning counts decreased significantly in irradiation group (P<0.05). (3)Compared with irradiation group, the number of platelets [(619.0±60.9)×10⁹/L vs (801.9±81.2)×10⁹/L] and the frequency of erythroid cells and myeloid cells decreased; moreover, hematopoietic colony forming units and single-cell cloning counts decreased significantly in irradiation+iron group (P<0.05). (4)Compared with irradiation group, ROS level increased by 1.94 fold in BMMNC, 1.93 fold in erythroid cells and 2.70 fold in myeloid cells, respectively (P<0.05).

CONCLUSIONThe dose of 4 Gy total body irradiation caused bone marrow damage and iron overload based on this injury model, which could damage bone marrow hematopoietic function aggravatingly. And further study found that iron overload was closely related to increased ROS level in BMMNC. The findings would be helpful to further study the injury mechanism of iron overload on the hematopoiesis of bone marrow.

In this study, we found that when Pseudomonas mendocina NK-01 accumulated intracellular carbon reserve, medium chain length poly (3-hydroxyalkanoates), it also synthesized extracellular saccharides, alginate oligosaccharides. The high carbon nitrogen ratio of culture medium facilitated alginate oligosaccharides production. We analyzed the structure of alginate oligosaccharide by Ultraviolet-Visible Spectrophotometry, Fourier Transform Infrared Spectroscopy, 1H and 13C of Nuclear Magnetic Resonance, and found that it was compounded in line from beta-D-mannuronic acids and alpha-L-gluronic acids via beta-(1-->4)/ alpha-(1-->4) bonds, which acetylated partly on the 2- and/or 3-hydroxy. In addition, we determined the weight-average molecular weight of alginate oligosaccharides by gel permeation chromatography to be 2054.
Alginates ; chemistry ; Glucuronic Acid ; biosynthesis ; chemistry ; genetics ; Hexuronic Acids ; chemistry ; Molecular Weight ; Oligosaccharides ; biosynthesis ; chemistry ; genetics ; Pseudomonas mendocina ; metabolism

Aim To investigate the influence of intranasal delivery of calcitonin gene-related peptide(CGRP)on cerebral blood supply and expression of vascular endothelial growth factor(VEGF)following experimental subarachnoid hemorrhage(SAH).Methods Wistar rats were divided into normal control group,SAH group,intranasal normal saline(NS)+SAH group and intranasal CGRP+SAH group.SAH models were produced by double injection of autologous arterial blood into cisterna magna.CGRP and NS were given by intranasal perfusion.Dynamic observations of regional cerebral blood flow(rCBF)of cerebral cortex were made using a laser Doppler flowmeter probe.On the third day after the second cisternal injection,the expression of VEGF protein in cerebral cortex was observed by immunofluorescence method combined with laser confocal microscopic observation.Results Anatomic observation revealed that SAH models were successfully manufactured.In SAH and intranasal NS+SAH groups,a drastic and persistent drop in rCBF was noted during the observed periods.The decrease of rCBF in intranasal CGRP+SAH group was slighter as compared with that in SAH and intranasal NS+SAH groups.In SAH and intranasal NS+SAH groups,increased expression of VEGF protein in cerebral cortex was observed on the third day after second cisternal injection as compared with that in normal control group.The expression of VEGF in intranasal CGRP+SAH group was more obvious than that in intranasal NS+SAH group.Conclusion Intranasal delivery of CGRP improves cerebral blood supply and promotes angiogenesis by enhancing the expression of VEGF after SAH.