Objective:To validate the efficacy of chimeric antigen receptor T cells targeting CD7 (CD7 CAR-T cells) modified with protein blocking technology and analyze whether pretreatment with dasatinib can enhance CD7 CAR-T killing ability or reverse the depletion phenotype.Methods:Green fluorescent protein (GFP)-labeled tumor cells were co-incubated with CD7 CAR-T cells or T cells at different potency-to-target ratios, but the culture volume and the numbers of CAR-T/T cells were same. The number of tumor cells was detected using flow cytometry. The killing effect of CAR-T cells on tumor cells was evaluated. A mouse model of acute T-lymphoblastic leukemia (T-ALL) was constructed by injecting 1×10 6 luciferase-expressing CCRF-CEM cells into the mouse tail vein to evaluate the therapeutic effect of CD7 CAR-T cells. Results:CD7 CAR-T cells had a significant killing effect on CCRF-CEM and Jurkat cells, but not on CD7-negative NALM6 cells. The mice in the group receiving CD7 CAR-T cells had a significantly reduced in vivo tumor load and a significantly prolonged survival time as compared with the mice in the group receiving untransduced T cells ( P<0.05). Dasatinib pretreatment significantly reversed the depletion phenotype of CD7 CAR-T cells ( P<0.05) and had no adverse effects on the killing effect and the proliferation of the cells. Conclusions:Protein-blocking technology-modified CD7 CAR-T cells are protected from killing each other, and pretreatment with dasatinib is expected to improve the efficacy and durability of CD7 CAR-T cells.

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 analyze the clinical characteristics of patients with novel coronavirus pneumonia (COVID-19) and the factors influencing mild cases developing into severe cases, so as to provide a basis for clinical screening, prevention and treatment of potential severe cases.Methods:Retrospective analysis was performed on the clinical characteristics of 168 cases who were admitted to two tertiary general hospitals in Anhui province and diagnosed with COVID-19 from January 20 to March 4, 2020. According to the classification criteria in the COVID-19 diagnosis and treatment program (trial version 6) issued by the National Health Commission, the mild and common cases were classified as the mild group ( n=137), and the severe and critical cases were classified as the severe group ( n=31). The general data, epidemiological history, clinical manifestations, laboratory examination and imaging indexes of the two groups were compared. Univariate analysis was performed. Then multivariate Logistic regression analysis was conducted on the factors with statistically significant differences in univariate analysis to obtain independent influencing factors of the occurrence of severe COVID-19. Results:Among the 168 COVID-19 patients, 95 were male and 73 were female, with an average age of 42.6±15.8 years old. The mean age of the mild group was younger than that of the severe group (40.5±15.5 vs 51.6 ±14.1, P<0.01). The proportion of patients combined with hypertension (29.0% vs 10.9%), diabetes (25.8% vs 2.2%, P=0.005) and two or more underlying diseases (29.0% vs 4.4%, P=0.006) in the severe group were significantly higher than those in the mild group. In the severe group, the proportion of patients receiving initial treatment in Medical institutions below secondary hospitals was significantly higher than that in the mild group ( P<0.01), and the time between symptom onset and diagnosis was longer [(8.00±3.27) d vs (6.49±3.90) d, P=0.048]. There was no significant difference in the initial symptoms between the mild group and the severe group. However, the body temperature was higher in the severe group [(38.80±0.67)℃ vs (37.9±0.60)℃, P<0.01]. At the time of admission, the lymphocyte percentage of the severe group was significantly lower than that of the mild group [(18.20±9.13)% vs (24.43±10.43)%, P<0.01], while C-reactive protein, interleukin-6 (IL-6), D-dimer, LDH, aspartate and aminotransferase were significantly higher than that of the mild group ( P<0.01). CT imaging showed that 11 (8%) patients in the mild group had lesions confined to a single lobe of the lung, while all patients in the severe group had multi-lobe lesions ( P<0.01). All the 168 COVID-19 patients in this study were cured, and the length of hospital stay in the severe group was significantly longer than that in the mild group [(24.71±7.72) d vs (20.28±7.67) d, P=0.021]. According to multivariate binary Logistic regression analysis, age ( P=0.042), diabetes ( P=0.021), body temperature at admission ( P=0.001), and IL-6 measured at admission ( P=0.008) were independent factors affecting COVID-19 to severe progress. Conclusions:Strengthening the professional knowledge training of primary hospitals is helpful for early diagnosis of COVID-19. Patients with older age, combined with diabetes, high initial fever and significantly increased IL-6 level are more possibly to develop into severe disease. Early identification and prevention should be carried out.

Objective:To analyze the clinical characteristics of critical patients with coronavirus disease 2019 (COVID-19), build an early warning model for severe/critical type, and aim at providing reference for the prediction of severe/critical COVID-19.Methods:The clinical data of COVID-19 patients treated in the Second People' Hospital of Fuyang City from January 20th to February 18th in 2020 were retrospective analyzed, including the demographic and epidemiological date, vital signs and hematology indexes, etc. on admission. Patients were divided into the normal type (set as normal group) and severe/critical type (set as severe group) according to the COVID-19 treatment plan classification standard published by National Health Commission of the People's Republic of China. The differences between two groups were compared, and the variables with statistical significance were incorporated in the multivariate binary unconditional Logistic regression analysis to screen the risk factors of severe/critical type. Risk factors were summarized to establish an early warning model, and the receiver operating characteristic (ROC) curve was carried out to evaluate the significance of the early warning model in the screening of critically COVID-19.Results:A total of 155 patients with COVID-19 were admitted, including 125 patients of normal type and 30 patients of severe/critical type. ① Compared with normal group, patients in severe group were older, and with higher proportion of basic diseases, higher body mass index (BMI), higher incidence of tachypnea, persistent high fever, peripheral blood oxygen saturation (SpO 2) < 0.95, while the white blood cell count (WBC), CD4 + T lymphocyte, CD8 + T lymphocyte, lymphocyte count (LYM) were decreased obviously, the levels of interleukin-6 (IL-6), C-reactive protein (CRP) and serum amyloid a protein (SAA), and CT showed higher incidence of multi-pulmonary lobe lesions. There were no significant differences of gender, travel history from Wuhan, smoking history, shock index (SI) and CD4 +/CD8 + ratio between the two groups. ② Multivariate Logistic regression analysis showed that age ≥60 years old [odds ratio ( OR) = 1.620, P = 0.031], combined with underlying diseases ( OR = 1.521, P = 0.044), persistent high fever ( OR = 2.469, P = 0.014), WBC < 2.0×10 9/L and/or LYM < 0.4×10 9/L ( OR = 3.079, P = 0.006), pulmonary multilobar lesions ( OR = 1.367, P = 0.047), and IL-6 ≥ 30 ng/L ( OR = 2.426, P = 0.010) were the risk factors of severe/critical COVID-19. ③ The OR value corresponding to each risk factors were scored by rounding. Two points were scored for age≥60 years old, with underlying diseases, persistent high fever and IL-6 ≥ 30 ng/L, 3 points for WBC < 2.0×10 9/L and/or LYM < 0.4×10 9/L, 1 point for pulmonary multilobar lesions, and totally calculated as early warning model scores. The early warning model score of the severe group was significantly higher than that of the normal group (9.33±2.79 vs. 5.04±2.38, t = 9.010, P = 0.001). ④ The ROC curve analysis showed the area under ROC curve (AUC) of early warning model on the early screening of severe/critical patients in COVID-19 was 0.944, and 95% confidence interval (95% CI) was 0.903-0.985; and the sensitivity and specificity were 93.3% and 72.0% respectively while the cut-off was 6.5. Conclusions:There are many differences between severe/critical and mild COVID-19 patients. The establishment of early warning model could help to screen severe/critical patients at an early stage, with certain significance for guiding treatment.

Objective:Retrospective analysis of the occurrence, development, treatment and outcome of three cases of pure red cell aplasia(PRCA) caused by parvovirus B19 after solid organ transplantation and literature review.Methods:Three patients were diagnosed by typical clinical manifestations, laboratory tests, and bone marrow aspiration. The parvovirus DNA was found to be positive in all 3 patients, and case 3 was found to be co-infected with herpes zoster virus. The treatment included a combination of immunosuppressive agents, intravenous gamma globulin, antiviral, hematopoietic, and hematopoietic materials supplementation.Results:The patient′s anemia was corrected, and the viral DNA became negative. No recurrence was found during follow-up.Conclusions:The etiological diagnosis of PRCA after solid organ transplantation is crucial, and comprehensive treatment can have a good effect.

Objective To investigate the clinical efficacy of arthroscopic double Endobutton plate internal fixation in the treatment of acute acromioclavicular joint dislocation.Methods A retrospective case series study was performed to analyze the clinical data of 32 patients with Rockwood type Ⅲ or Ⅴ acute acromioclavicular joint dislocation admitted to Foshan Hospital of TCM from May 2015 to February 2018.There were 26 males and six females,aged 22-56 years [(37.7 ± 1.6)years].There were 22 patients with Rockwood type Ⅲ and 10 patients with type Ⅴ.The duration from injury to operation was 2-5 days.All patients were treated with arthroscopic double Endobutton plate internal fixation for coracoclavicular ligament reconstruction.The operation time,hospitalization time,and postoperative shoulder joint braking time were recorded.The shoulder joint function was evaluated by the Constant-Murley score (CMS),visual analogue scale (VAS),American shoulder and elbow surgeons (ASES) score,University of California Los Angeles (UCLA) score and simple shoulder test (SST) before operation and at the last follow-up.Meanwhile,complications such as infection,loosening and joint dislocation were recorded.Results All patients were followed up for 8-48 months [(27.8 ± 11.6) months].The operation time was 48-93 minutes [(65.0 ± 24.6) minutes],the hospitalization time was 3.2-10 days [(6.5 ± 2.6)days],and the postoperative braking time was 21-35.3 days [(28.2 ± 7.1)days].All patients had a grade A healing of surgical incision.The VAS was decreased from preoperative (5.7 ±2.5)points to (1.2 ±0.8)points at the last follow-up (P ＜0.01);CMS wasvincreased from preoperative (29.3 ± 3.4) points to (93.2 ± 2.1) points at the last follow-up (P ＜0.05);ASES score was increased from preoperative (28.9 ± 10.1) points to (95.7 ± 5.6) points at the last follow-up (P ＜ 0.05);UCLA score was improved from preoperative (11.6 ± 2.8) points to (34.2 ±1.5)points at the last follow-up (P ＜0.05);SST score was improved from preoperative (3.5 ±1.2)points to (9.7± 1.4)points at the last follow-up (P ＜ 0.O1).The incidence of intraoperative and postoperative complications was 16％ (5/32).In two patients,the surgery was changed to U shape fixation circulating the coracoid base due to the coracoidbone tunnel cutting out.One patient was found to have insufficient reduction and received the acromioclavicular joint operation.One patient had reduction loss due to the clavicle button collapsing,and one patient had excessive reduction.Conclusions For Rockwood type Ⅲ or Ⅴ acute acromioclavicular dislocation patients,arthroscopic double Endobutton plates for coracoclavicular ligament reconstruction has the advantages of less intraoperative trauma,quick recovery and satisfactory clinical result.However,the incidence of intraoperative and postoperative complications is relatively high,and indications should be strictly controlled.

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 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.

Objective: To evaluate the feasibility and potential value of comprehensive geriatric assessment (CGA) in elderly (≥60 years) patients with newly diagnosed acute myeloid leukemia (AML) in China. Methods: The CGA results of 83 newly diagnosed AML (non-APL) patients from 16 hospitals in Beijing and Tianjin between March 2016 and December 2017 were prospectively collected and analyzed. The clinical data, treatment and follow-up information were also collected. Results: Of 83 newly diagnosed elderly AML patients, 81 patients (97.6%) completed all designated CGA assessment. The median number of impaired scales of the CGA assessment in the studied population was 2(0-6). Sixteen patients (19.3%) showed no impairments according to the geriatric assessment scales implem ented by this study. The distributions of impaired scales were as follows: impairment in ADL, 55.4%; IADL impairment, 42.2%; MNA-SF impairment, 48.2%; cognitive impairment, 15.7%; GDS impairment, 31.7%; HCT-CI impairment, 19.5%, respectively. In patients with "good" ECOG (n=46), the proportion of impairment for each CGA scale ranged from 6.5% to 37.0% and 32 patients (68.9%) had at least one impaired CGA scale. Survival analysis showed that the number of impaired scales of the CGA was significantly correlated with median overall survival (P=0.050). Conclusions CGA was a tool with feasibility for the comprehensive evaluation in elderly AML patients in China. Combined with age and ECOG, CGA may be more comprehensive in assessing patients’ physical condition.

Objective: To explore the clinical characteristics, diagnosis, treatment and prognosis of patients with hematological diseases that complicated by mucormycosis, and to improve the understanding and clinical diagnosis and treatment of the disease. Methods: The clinical data of 7 patients suffering from mucormycosis during September 2012 and September 2018 were retrospectively analyzed, and their clinical characteristics, treatment process and prognosis were analyzed. Results: Of 7 patients, there were 4 males and 3 females, with a median age of 36 (19-79) years old. Two patients were diagnosed as acute myeloid leukemia as the underlying disease, the other 5 patients suffered from acute B lymphoblastic leukemia, peripheral T cell lymphoma, chronic myelocytic leukemia in blastic phase, myeloproliferative neoplasm and severe aplastic anemia after transplantation, respectively. Among them, disease types of mucormycosis were pulmonary in 4 patients, rhino-orbital-cerebral in 1 patient, cutaneous in 1 patient and disseminated in 1 patient. All the cases were confirmed by biopsy histopathology. The treatment drugs were amphotericin B or liposomal amphotericin B, and posaconazole. Surgical treatment was performed in 4 patients, 3 out of 4 achieved radical debridement, and the other one had local debridement. Two patients were cured, 1 patient was improved and 4 patients died. Conclusions The clinical manifestation and image feature of mucormycosis in patients with hematological diseases were diverse, and the mortality rate is high, diagnosis mainly depends on histopathology. Early diagnosis, control of underlying disease, improvement of immunosuppressive status, timely effective antifungal therapy and radical surgical debridement are the key points for improving the survival rate of patients with hematological diseases complicated by mucormycosis.