Objective To explore the phenotypic conversion of regulatory T cells (Tregs) in the lungs of mice with bronchopulmonary dysplasia (BPD)-affected mice. Methods A total of 20 newborn C57BL/6 mice were divided into air group and hyperoxia group, with 10 mice in each group. The BPD model was established by exposing the newborn mice to hyperoxia. Lung tissues from five mice in each group were collected on postnatal days 7 and 14, respectively. Histopathological changes of the lung tissues was detected by HE staining. The expression level of surfactant protein C (SP-C) in the lung tissues was examined by Western blot analysis. Flow cytometry was performed to assess the proportion of FOXP3⁺ Tregs and RORγt⁺FOXP3⁺ Tregs in CD4⁺ lymphocytes. The concentrations of interleukin-17A (IL-17A) and IL-6 in lung homogenate were measured by using ELISA. Spearman correlation analysis was used to analyze the correlation between FOXP3⁺Treg and the expression of SP-C and the correlation between RORγt⁺FOXP3⁺ Tregs and the content of IL-17A and IL-6. Results The hyperoxia group exhibited significantly decreased levels of SP-C and radical alveolar counts in comparison to the control group. The proportion of FOXP3⁺Tregs was reduced and that of RORγt⁺FOXP3⁺Tregs was increased. IL-17A and IL-6 concentrations were significantly increased. SP-C was positively correlated with the expression level of RORγt⁺FOXP3⁺ Tregs. RORγt⁺FOXP3⁺ Tregs and IL-17A and IL-6 concentrations were also positively correlated. Conclusion The number of FOXP3⁺ Tregs in lung tissue of BPD mice is decreased and converted to RORγt⁺ FOXP3⁺ Tregs, which may be involved in hyperoxy-induced lung injury.
Animals ; Mice ; Mice, Inbred C57BL ; Bronchopulmonary Dysplasia ; T-Lymphocytes, Regulatory ; Interleukin-17 ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; Hyperoxia ; Interleukin-6 ; Forkhead Transcription Factors ; Lung

OBJECTIVE: The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model. METHODS: Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model. RESULTS: On the 10th day after inoculation, hCD45⁺ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3⁺ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks. CONCLUSION Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.
Humans ; Animals ; Mice ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Heterografts ; Bone Marrow ; Disease Models, Animal ; T-Lymphocytes ; Mice, SCID

OBJECTIVETo analyze the incidence of bone marrow involvement in patients with different pathological types of lymphoma.

METHODSThe results of bone marrow tests including bone marrow aspiration(BMA), flow cytometry detection, bone marrow biopsy(BMB) and F-FDG PET/CT, were analyzed retrospectively in 702 cases of newly diagnosed lymphoma with bone marrow assessment in our hospital from October 2000 to September 2016. If one of the above-mentioned 4 tests showed positive, the lymphoma patient was judged as bone marrow involved.

RESULTSThe incidence of bone marrow involvement (BMI ) in the patients with NHL was much higher than that in patients with HL [32.6 %(201/616) vs 15%(13/86)](P<0.05). For patients with NHL, the incidence of bone marrow involvement in B-cell lymphoma was higher than that in T-cell lymphoma (37.0% vs 22.6%)(P<0.05). According to different pathological types, the incidences of BMI in the patient with mantle cell lymphoma, hepatosplenic T-cell lymphoma, diffuse large B-cell lymphoma (DLBCL) and follical lymphoma (FL) were 88% (25/22), 100% (5/5), 21.8% (56/257), and 38.5% (15/39) , respectively.

CONCLUSIONThe incidence of bone marrow involvement varies in different pathological types of lymphoma.Bone marrow assessment has significant importance for stading of newly diagnosed lymphoma patients.

Biopsy ; Bone Marrow ; Fluorodeoxyglucose F18 ; Humans ; Incidence ; Lymphoma ; Positron Emission Tomography Computed Tomography ; Retrospective Studies

OBJECTIVETo explore the prognostic value of interimF-FDG PET/CT (i-PET/CT) scan for the patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL).

METHODSA total of 70 cases of initially diagnosed of DLBCL by 158F-FDG PET/CT scans in our hospital were retrospectively analyzed. The 5-point scale, the Lugano classification and maximum standardized uptake value induction (ΔSUVmax) criteria were used respectively to assess i-PET/CT scans. Receiver-operating characteristics (ROC) analysis was used to determine an optimal cutoff for ΔSUVmax. Progression-free survival (PFS) and overall survival (OS) times were estimated as prognostic indicators using the Kaplan-Meier method and Cox regression.

RESULTSOptimal cutoff to predict progression or death was 62% for ΔSUVmax. The positive predictive value (PPV) for 2-year PFS and OS of i-PET/CT diagnosed by 5-point scale was low, and could be improved by using the Lugano classification with decreased sensitivity or ΔSUVmax criteria. Kaplan-Meier survival curve analysis showed that the Lugano classification and ΔSUVmax were good predictors for PFS and OS, respectively, while the 5-point scale could only predict OS. Cox regression univariate analysis showed that the International Prognostic Index (IPI) score was better to predict PFS than 5-point scale, but worse than the three assessments in predicting OS. COX regression multivariate analysis showed that ΔSUVmax<62% was an independent risk factor of prognosis, while the Lugano classification was only the OS independent prognostic predictor.

CONCLUSIONAssessing i-PET/CT by 5-point scale is a limited value for predicting PFS and OS in DLBCL patients. The Lugano classification is recommended to discriminate the patients with poorer outcomes. The ΔSUVmax criteria for i-PET/CT of DLBCL patients is an independent prognostic predictor for PFS and OS, better than the IPI score.

OBJECTIVETo investigate the correlation between the bone marrow fibrous proliferation and the prognosis of acute myeloid leukemia(AML).

METHODSThe quantitative method was used to analyze the reticulin fiber density (RFD) of AML patients. the bone marrow sections from 39 primary AML patients and 35 normal controls were collected to compare the RFD between these 2 groups. The prognosis value of RFD for AML were estimated by using appropriate statistical analysis.

RESULTSRFD in primary AML was significantly higher than that in normal controls(2.41%±0.23% vs 1.14%±0.06%)(P<0.05). Relapse-free survival(RFS) analysis showed that the patients with RFD more than 1.68% indicated poor RFS, and the overall survival(OS) analysis showed that patients with RFD more than 2.66% indicated poor overall survival (P<0.05). Besides, there were no relationship between RFD and the BM blast count (r=0.01) and WBC counts (r=0.04) at diagnosis(P>0.05).

CONCLUSIONThe RFD in bone marrow is a high risk factor in poor prognosis of AML patients.

OBJECTIVETo isolate platelet-rich plasma(PRP) from the white slurry(WS), a depleted fraction of the clinical blood supply, so as to provide an easier method to harvest PRP for related studies and clinical use.

METHODSThe protocols preparing PRP from whole blood and WS were compared. The morphological characteristics of the different PRPs were observed under transmission electron microscope; the expression of the platelet markers CD41a and CD42b were detected by the flow cytometry. Moreover, the ingredients of the PRPs were measured by using cytoanalyzer. for detecting the physiological function of the PRP, the harvested PRP were added to MSC culture and the cell proliferation was detected by using CCK-8 method.

RESULTSa large amount of PRP from WS was easier harvested. the WS-derived PRP shared similar morphological characteristics and ingredients as compared with whole blood-derived PRP. Importantly, the WS-derived PRP exhibited a higher expression of CD41a and CD42b than that of traditional PRP, which indicate that the WS is a promising reservoir for PRP.

CONCLUSIONThe WS can be used to prepare PRP, and the novel PRP share similar biological characteristics as traditional PRP prepared from whole blood. The present study provides an easier and economical method to harvest PRP and this findings may be helpful for PRP related studies.

OBJECTIVETo explore the effects of simvastatin on vascular endothelial cell apoptosis and Bcl-2 protein expression in the aorta in a rat model of atherosclerosis.

METHODSThirty-six rats were randomized into control group (n=10), atherosclerosis model group (n=13) and simvastatin intervention group (n=13). In the latter two groups, rat models of atherosclerosis were established by intraperitoneal injection of vitamin D3 combined with high-fat feeding for 6 weeks, and the control rats were fed with regular diet. In the intervention group, the rats were further fed with high-fat diet with daily simvastatin treatment for 4 weeks. After the treatments, the pathological changes and plaque in the thoracic aorta were observed, and the expression of Bcl-2 protein was detected with immunohistochemistry. TUNEL assay was used to determine the apoptosis index (AI) of the vascular endothelial cells.

RESULTSCompared with that in the control group, Bcl-2 protein expression in the aorta of atherosclerotic rats was significantly decreased (P<0.05); simvastatin treatment obviously increased the expression of Bcl-2 protein in atherosclerotic rats (P<0.05) to a level similar to that in the control group. The AI was the highest in the model group (P<0.05) and comparable between the control and simvastatin treatment group.

CONCLUSIONThe therapeutic effect of simvastatin against atherosclerosis is probably mediated by up-regulation of Bcl-2 protein, which inhibits vascular endothelial cell apoptosis in rats with aortic atherosclerosis.

Objective To investigate the practice and effect of specialist nursing inspection of intravenous infusion in the quality control of specialist nursing. Methods Intravenous infusion treatment nursing group used feed-forward control theory and methods, taking cross-sectional research methods yearly from 2014 to 2016 and referencing intravenous infusion special quality checklist to fully understand the hospital infusion tool selection situation and quality. Intravenous infusion related treatment system operational procedures was established and improved, the monthly quality control plan was made to do quality nursing of intravenous infusion specialist inspections and timely feedback of each month to various sections of the hospital. Results The rational choice of the hospital infusion tool was more reasonable. The proportion of steel needle in 2016 compared with in 2015 and 2014 was decreased by 8.9%(321/920-212/846), 18.8%(321/920-156/968). Intravenous infusion quality was significantly improved in the catheters′normative fix, nursing staffs′mastery of the skills and related expertise control rate, intravenous infusion treatment-related complications e.g. Phlebitis above grade two in 2016 were reduced 22 cases and 30 cases compared with in 2015 and in 2014, meanwhile the number was 18 and 31 in drug extravasation case. Conclusions Special nursing of intravenous infusion has a good effect on the improvement of nursing quality, and feed-forward control is an effective method for quality control of intravenous infusion therapy.

OBJECTIVETo explore the clinical and prognostic features as well as treatment response of childhood B-cell non-Hodgkin's lymphoma/acute lymphoblastic leukemia (B-NHL/B-ALL), so as to better modify the treatment for further improving the prognosis.

METHODSThe clinical data of 43 patients with newly-diagnosed childhood B-NHL/B-ALL from July 2005 to December 2013 in West China Second Hospital of Sichuan University were retrospectively analyzed with particular focus on clinical presentations, laboratory findings and histology. Among them 26 patients received B-NHL-2010 protocol and 17 patients received LMB-89 protocol treatment. Kaplan-Meier method was used to compare the survival rates between groups, while multiple factor logistic regression was used to identify the prognostic factors.

RESULTS(1) The median age at diagnosis was 7.58 (2.42-13.67) years. The male-to-female ratio was 2.9 : 1. No significant difference was found in the median age at diagnosis between male and female children with B-NHL/B-ALL (P = 0.837). (2) Burkitt's lymphoma was the most common (34/43, 79.07%), followed by diffuse large B cell lymphoma (4/43, 9.3%), ALL-L3 (3/43, 6.98%) and others (2/43, 4.65%) in decreasing frequency. (3) According to St. Jude staging classification, 4 patients (9.30%) were divided into stage I, 9 patients (20.93%) into stage II, 23 patients (53.49%) into stage III and 7 patients (16.28%) into stage IV; (4) Clinically, the common predilection sites were as following: ileocecus (11/43, 25.58%), nasopharynx (10/43, 23.26%), faciomaxillary (9/43, 20.93%), superficial lymphadenopathy (8/43, 18.60%), other sites such as mediastinum and bone marrow (5/43, 11.63%). (5) With a median follow up of 24 months (0.7-105 months), the 2-year overall survival (OS) rate and event-free survival (EFS) rate were 79.8% ± 6.5%% and 71.0% ± 7.2%, respectively. The 2-year OS and EFS rates in patients treated with B-NHL-2010 protocol were 79.1% ± 8.4% and 74.1% ± 8.4%, while those in patients treated with LMB-89 protocol were 87.5% ± 8.3% and 66.7% ± 12.4%, respectively, but there was no significant difference between them (P > 0.05). The 2-year EFS rate in patients with LDH > 2N and bone marrow infiltration were significantly lower than that of other groups (P < 0.05). (6) 8 patients (18.6%) relapsed. The median relapsed time was 6 months (2-9 months). 1 patient suffered progressive disease. Male, systemic symptom, elevated LDH, bone marrow and CNS infiltration and advanced stage (stage III and stage IV) were associated with relapse /progressive disease. Logistic regression analysis showed that LDH > 2N was an independent unfavorable prognostic factors (OR = 31.129, P = 0.02).

CONCLUSIONOutcome of B-NHL/B-ALL is greatly improved by current intensive and short-time chemotherapy regimen. The 2-year event-free survival (EFS) rate is 71.0% ± 7.2%. There is no significant difference in EFS rate between patients treated with B-NHL-2010 protocol and LMB89 protocol. The long-term survival rate in patient with advanced disease need to be further improved.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Burkitt Lymphoma ; diagnosis ; drug therapy ; Child ; Cyclophosphamide ; therapeutic use ; Cytarabine ; therapeutic use ; Disease-Free Survival ; Doxorubicin ; therapeutic use ; Etoposide ; therapeutic use ; Female ; Humans ; Hydrocortisone ; therapeutic use ; Leucovorin ; therapeutic use ; Logistic Models ; Lymphoma, B-Cell ; diagnosis ; drug therapy ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; drug therapy ; Male ; Methotrexate ; therapeutic use ; Multivariate Analysis ; Neoplasm Staging ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; drug therapy ; Prednisone ; therapeutic use ; Prognosis ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use

OBJECTIVETo study the relationship between surface markers of CD56 and CD19 and karyotypes and prognosis in multiple myeloma.

METHODSA total of 126 cases of newly diagnosed multiple myeloma in the first hospital of Peking university from 2011 to 2015 were enrolled in this study. Cytogenetic abnormalities and immunophenotypes were detected by using fluorescence in situ hybridization and flow cytometry respectively before chemotherapy. Bone marrow smear was used for detection of abnormal plasma cell infiltration. By combining with their basic data, the relationship between immunophenotypes, cytogenetics and prognosis of MM was analyzed.

RESULTS(1) The median of myeloma cells in the 126 patients was 0.24（0.01-0.97）; the median of myeloma cells in 116 patients who have immunophenotype datas was 0.25（0.01-0.97）； the median of myeloma cells in CD19 positive patients was 0.11（0.01-0.53）; the median of myeloma cells in CD19 negative patients was 0.26（0.01-0.97）. The median of myeloma cells in CD19 positive patients was much lower than that in CD19 negative patients（P=0.036）. (2)In 116 patients detected by the immunophenotype, the myeloma cells expressed CD19,CD20,CD56 and CD117. Compared with CD56 negative patients(45/116,38.79%),CD56 positive patients(71/116,61.21%) had a clearly favorable disease outcome（OS was 53.0 month vs 31.0 month,P=0.016; PFS was 37.5 months vs 18.4 months, P=0.036）. (3)CD19 positive patients was 16.38%（19/116）,CD19 negative patients was 83.62%（97/116）； CD19 positive MM and CD19 negative MM had no difference in OS and PFS. (4)CD117 positive rate in CD19 positive patients was 42.11%(8/19), the CD117 positive rate in CD19 negative patients was 18.57%(18/97), the CD19 expression positively correlated with CD117 expression. (5)FISH detection was done for 67 newly diagnosed MM patients, 8 patients showed normal karyotypes(11.94%), 59 patients had abnormal karyotypes(88.06%). The most common abnormal karyotypes were IgH rearragement which occurred in 47 patients(70.15%). Other abnormal karyotypes included 1q21+, del(13q14),del(13q14.3),del(17p13) . These abnormal karyotypes occurred in 37 patients（55.22%）,31 patients（46.27%）,33 patients（49.25%） and 13 patients（19.40%） respectively. In comparison with CD19 negative MM patients, the incidence rate of 1q21+ and del(13q14.3) was significantly lower in CD19 positive patients(1q21+:33.33% vs 61.54%,P=0.016; del(13q14.3): 33.33% vs 53.85%,P=0.043）.

CONCLUSIONThe prognosis of CD56 positive MM patients is better than that of CD56 negative MM patients, CD19 negative MM has more abnormal karyotypes and bone marrow infiltration,but they have no statistical prognostic differences.

Chromosome Aberrations ; Chromosome Deletion ; Flow Cytometry ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Karyotyping ; Multiple Myeloma ; Prognosis