OBJECTIVE: To understand the biological characteristics of polycythemia vera (PV) patients with myeloid fibroplasia, and further analyze the risk factors affecting myeloid fibroplasia in PV patients, so as to provide ideas for predicting the occurrence of myeloid fibroplasia in PV patients. METHODS: Forty patients with PV in the Department of Hematology, Xiyuan Hospital of China Academy of Chinese Medical Sciences were collected and divided into two groups, with (hyperplasia group) and without (Non-proliferative group) hyperplasia of bone marrow fibers. The differences of basic clinical characteristics, blood routine, biochemistry, bone marrow cells, coagulation function and other indicators between the two groups were compared, and the independent risk factors affecting the proliferation of bone marrow fibrous tissue in PV patients were further analyzed by multivariate regression. RESULTS: Compared with Non-proliferative group, the JAK2 mutation rate (95% vs 70%，P=0.037), eosinophilic cell count (0.19 vs 0.11, P=0.047) and eosinophilic percentage (1.84 vs 1.27, P=0.001) in PV patients with hyperplasia were significantly increased, triglycerides (1.55 vs 1.91, P=0.038) and low-density lipoprotein (1.50 vs 3.08, P=0.000) were significantly reduced, bone marrow hematopoietic volume (0.85 vs 0.6, P=0.001), granulocyte/erythrocyte ratio (3.40 vs 1.89, P=0.033), lymphocyte/erythrocyte ratio (0.60 vs 0.42, P=0.033), and granulocyte+lymphocyte/erythrocyte ratio (3.72 vs 2.37, P=0.026) were significantly increased, thrombin time (18.84 vs 18.12, P=0.043) was significantly prolonged. Multivariate regression analysis results showed that peripheral blood eosinophil ≥2% and low-density lipoprotein ≤2 mmol/L were independent risk factors for bone marrow fibrous tissue hyperplasia in PV patients (P<0.05). CONCLUSION Increased proportion of peripheral blood eosinophils and decreased low density lipoprotein are risk factors for bone marrow fibrous tissue hyperplasia in PV patients.
Humans ; Bone Marrow/pathology* ; Polycythemia Vera ; Hyperplasia/pathology* ; Granulocytes/pathology* ; Janus Kinase 2/genetics* ; Risk Factors ; Lipoproteins, LDL ; Polycythemia/pathology*

OBJECTIVE: To investigate the significance of peripheral blood lymphocyte to monocyte ratio (LMR) and corrected levels of serum calcium (cCa) as prognostic markers for the newly diagnosed multiple myeloma (MM) patients. METHODS: The clinical data of 114 newly diagnosed MM patients in the Second Affiliated Hospital of Kunming Medical University from January 2013 to March 2020 were retrospectively analyzed. Receiver operating characteristic (ROC) curve analysis was used to identify the optimal cutoff value, and the patients were divided into high LMR group and low LMR group (LMR≥3.35 and LMR < 3.35). Moreover, the patients were divided into four groups according to initial diagnosis LMR and LMR after four courses of treatment (LMR4): Group A (LMR≥3.35, LMR4≥3.35), Group B (LMR≥3.35, LMR4 < 3.35), Group C (LMR < 3.35, LMR4≥3.35), and group D (LMR < 3.35, LMR4 < 3.35). The simple prognosis model was established by combined with LMR and cCa, the patients were divided into Group a (no risk factor), group b (1 risk factor) and Group c (2 risk factors). Independent sample T-test, Pearson Chi-square test or Mann-Whitney U test were used to evaluate the differences between various parameters, and Kaplan-Meier method and Cox regression were used for survival analysis. RESULTS: The median follow-up time was 13.05（0.1-72.5）months. Survival analysis showed that the patients with low LMR predicted poor prognosis, the overall survival (OS) time of the patients with low LMR was significantly shorter (17 vs 50.5 months, P=0.006) than the patients with high LMR, the difference was also significant between group A and Group D (56.5 vs 30.5 months, P=0.043). The OS of the patients was also significantly shorter in the high cCa group (≥2.75 mmol/L) compared with normal group (8.5 vs 34 months, P=0.006). Multivariate survival analysis showed that LMR < 3.35 (P=0.028) and cCa≥2.75 mmol/L (P=0.036) were the independent risk factors affecting prognosis of MM patients. The comparison of risk factors showed that the median OS of Group a, b and c was 50, 20, and 8.5 months, respectively. The prognosis of the patients without risk factors was better than that of patients with 1-2 risk factors (Group a vs Group b, P < 0.0001; Group a vs Group c, P=0.002). CONCLUSION LMR and cCa are the independent risk factors affecting the prognosis of newly diagnosed MM patients, and the development of a simple prognosis system combining them can quickly identify the prognosis of newly diagnosed MM patients.
Calcium ; Humans ; Lymphocytes ; Monocytes ; Multiple Myeloma ; Prognosis ; Retrospective Studies

Calreticulin/genetics* ; DNA Methylation ; Female ; Humans ; Janus Kinase 2/genetics* ; Mutation ; Myeloproliferative Disorders/genetics* ; Polycythemia Vera/genetics*

OBJECTIVE: To analysis the specific protein markers of essential thrombocythemia (ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation. METHODS: Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P＜0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P＜0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring (PRM) technology. RESULTS: A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing. CONCLUSION Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV, GPVI, COL1A1 and GP1bα can be used as new targets related to ET platelet activation.
Blood Platelets/metabolism* ; Humans ; Platelet Activation ; Platelet Membrane Glycoproteins/metabolism* ; Technology ; Thrombocythemia, Essential

OBJECTIVE: To explore and design a novel bi-specific chimeric antigen receptor (CAR) structure. To obtain the corresponding CAR-T cells and verify killing effects on tumor cells in vitro and in vivo. METHODS: Five kinds of bi-specific CAR structures including humanized CD19 scFv and CD79b scFv, CD8 hinge & TM-4-1BB-CD3ζ and/or CD3ε chain intracellular regions were constructed and prepared. CAR-19-79b cells were obtained. Five kinds of CAR-T cells were co-incubated with the 3M-CD19-CD79b-Luc target cells. Luciferase assay and ELISA were used to detecte the killing ability of these five groups of CAR-T cells and the secretion of cytokines and compared. The optimal structure of CAR-T cells was used to treat the leukemia mouse model constructed by Daudi-Luc cells. And the treatment efficacy was evaluated. At the same time, other targets were used in this structure. With the same methods, the stability and effectiveness of the structure were verified. RESULTS: CAR-19-79b-T cells were cultured for 7 days, the expression rates of CAR-19 and CAR-79b were 21.6%-36.3% and 21.7%-37.8%, respectively. The killing rates of 5 kinds of CAR-19-79b-T cells prepared by T cells from 3 healthy donors on 3M-CD19-CD79b-Luc cells were significantly higher than those of the T cell control group at the effect-target ratio of 10∶1. Among them, the killing rates of CAR-19-79b-T cells with No. III and No. IV structures were the strongest. After co-incubation with 3M-CD19-CD79b-Luc target cells, the amount of IFN-γ and TNF-α secreted by CAR-T cells with CAR IV and CARV structures was the lowest. And there was no significance between the two groups (P>0.05). CAR IV cells with remarkable killing effect and low secretion factor had obvious therapeutic effect on Daudi-Luc leukemia mice, extending the survival period of mice to 64 days. And all mice in the T cell control group died at 41.0±2.4 days. The CAR-19-BCMA-T and CAR-19-22-T with the same structure showed significant killing ability and low cytokine expression levels. CONCLUSION A novel bi-specific CAR structures was successfully designed, which could efficiently kill the corresponding tumor cells and secrete less cytokines (such as TNF-α, IFN-γ). Moreover, it shows obvious therapeutic effect on Daudi lymphoma mouse model. The bi-specific CAR structure shows good killing specificity and safety.
Animals ; Mice ; Leukemia ; Receptors, Chimeric Antigen ; T-Lymphocytes ; Tumor Necrosis Factor-alpha

OBJECTIVE: To investigate the distribution and drug resistance of nosocomial infection pathogens in AL patients with hematological agranulocytosis, so as to provide evidence for the clinical rational use of antibiotics. METHODS: Pathogenic data of 504 hospitalized patients with agranulocytosis caused by nosocomial infection in the Department of Hematology, the First Hospital of Lanzhou University from May 2015 to May 2018 were collected and retrospectively analyzed for the distribution of pathogenic bacteria and the results of drug susceptibility. RESULTS: The isolated pathogenic bacteria strains amounted to 184, out of which, 168 strains (91.3%) orginated from the patients with acute leukemia, while 16 strains (8.7%) originated from the patients with non-acute leukemia. The positive samples mainly originated from blood stream, the isolated bacteria from which were 81 straims (44%); then originated from sputam and pharynx swabs, from which isolated bacteria amounted to 54 strains (29.3%) and 35 strains (19%) respectively. In the pathogenic bacteria, the Gram-negative bacteria amounted to 126 strains accounting for 68.46%, out of which the most commond bacteria strains were Klebseilla pneumoniae, cscherichia coli and Pseudomonas aeruginosa; the Gram positive bocteria amounted to 23 strains accounting for 12.5%, mainly staphy lococeus anreus, and Staphylococcus epitermidis; the fungi amounted to 35 strains accounting for 19.02%, mainly Candida albicans. The detection rates of Escherichia coli and Klebsiella pneumoniae producing extended-spectrum β-lactamases (ESBLs) were 40.0% and 22.2%, respectively. They were 100% sensitive to amikacin and 27.8% resistant to carbapenems. Klebsiella pneumoniae had the highest sensitivity to amikacin, 94.44% to ampicillin, 97.22% to carbapenems and 100% sensitive to ammonia. Their penicillin-resistance rate was the highest, up to 80%; Pseudomonas aeruginosa was sensitive to the antibiotics (＞80%）. Methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative Staphylococcus were detected in Gram-positive bacteria. The susceptibility rate of main Gram-positive bacteria to vancomycin and linezolid was 100%, and they were 100% resistant to penicillin. CONCLUSION Gram-negative bacteria are the main pathogens of nosocomial infection in patients with hematological agranulocytosis. Pathogens have different resistance to antimicrobial agents. It is important to know the distribution and susceptibility of common pathogens for rational selection of antimicrobial agents and control of nosocomial infection.
Cross Infection ; Drug Resistance ; Drug Resistance, Bacterial ; Gram-Negative Bacteria ; Humans ; Methicillin-Resistant Staphylococcus aureus ; Microbial Sensitivity Tests ; Retrospective Studies

BACKGROUND AND OBJECTIVE: Acute liver failure (ALF) is a type of disease with high mortality and rapid progression with no specific treatment methods currently available. Glucocorticoids exert beneficial clinical effects on therapy for ALF. However, the mechanism of this effect remains unclear and when to use glucocorticoids in patients with ALF is difficult to determine. The purpose of this study was to investigate the specific immunological mechanism of dexamethasone (Dex) on treatment of ALF induced by lipopolysaccharide (LPS)/D-galactosamine (D-GaIN) in mice. METHODS: Male C57BL/6 mice were given LPS and D-GaIN by intraperitoneal injection to establish an animal model of ALF. Dex was administrated to these mice and its therapeutic effect was observed. Hematoxylin and eosin (H&E) staining was used to determine liver pathology. Multicolor flow cytometry, cytometric bead array (CBA) method, and next-generation sequencing were performed to detect changes of messenger RNA (mRNA) in immune cells, cytokines, and Kupffer cells, respectively. RESULTS: A mouse model of ALF can be constructed successfully using LPS/D-GaIN, which causes a cytokine storm in early disease progression. Innate immune cells change markedly with progression of liver failure. Earlier use of Dex, at 0 h rather than 1 h, could significantly improve the progression of ALF induced by LPS/D-GaIN in mice. Numbers of innate immune cells, especially Kupffer cells and neutrophils, increased significantly in the Dex-treated group. In vivo experiments indicated that the therapeutic effect of Dex is exerted mainly via the glucocorticoid receptor (Gr). Sequencing of Kupffer cells revealed that Dex could increase mRNA transcription level of nuclear receptor subfamily 4 group A member 1 (Nr4a1), and that this effect disappeared after Gr inhibition. CONCLUSIONS In LPS/D-GaIN-induced ALF mice, early administration of Dex improved ALF by increasing the numbers of innate immune cells, especially Kupffer cells and neutrophils. Gr-dependent Nr4a1 upregulation in Kupffer cells may be an important ALF effect regulated by Dex in this process.
Animals ; Dexamethasone/therapeutic use* ; Disease Models, Animal ; Kupffer Cells/physiology* ; Liver Failure, Acute/pathology* ; Male ; Mice ; Mice, Inbred C57BL ; Nuclear Receptor Subfamily 4, Group A, Member 1/physiology* ; Receptors, Glucocorticoid/physiology*

Trauma is one of the leading causes of death worldwide. It is an urgent task to strengthen the trauma care and prevent the complications. In 2018, Chinese Journal of Traumatology reported a series of trauma-related articles of which the contents include pre-hospital care, in-hospital care and complication prevention, et al, aiming to improve the treatment levels, decrease the trauma incidence, and reduce the trauma mortality and disability.
China ; Humans ; Periodicals as Topic ; Societies, Medical ; organization & administration ; Time Factors ; Traumatology ; organization & administration ; Wounds and Injuries ; prevention & control ; therapy

OBJECTIVE: To explore the values of 4 prognostic score systems in evaluation of clinical effecacy for patients with newly diagnosed chronic myeloid leukemia in chronic phase (CML-CP) treated with imatnib mesylate (IM) and the relationship between 4 prognostic score systems and deep molecular response (MR4.5). METHODS: The clinical data of 240 CML-CP patients treated with imatinib mesylate in our hospital between Janunay 2008 and December 2017 were analyzed retrospecively. The risk was stratified according to 4 prognostic score systems, the relationship between the 4 prognostic score systems and 3-month early molecular response (3M-EMR), 6 month complete cytogenetic response (6M-CCyR), 12-month major molecular response (12M-MMR) as well as the correlation of the 4 prognostic score systems with deep molecalar response were analyzed. RESULTS: At the end of treatment for 3 months, the EMR was evaluated for 219 patients, among them 164 (74.9%) patients achieved 3M-EMR; at the end of treatment for 6 months, CCyR was evaluated for 180 pathsents, among them 130 (72.2%) patients achicved 6M-CCyR; at the end of treatment for 12 months, the MMR was evaluated for 111 patients, among them 60 (54.1%) patients achieved 12M-MMR. Compared with the high-risk group, the treatment response to IM in the low-risk group (including the low-risk group and the intermediate-risk group) was better. There was significant difference in 3M-EMR according to Sokal score and ELTS score (P＜0.05), and there was significant difference in 12M-MMR according to EUTOS score and ELTS score (P＜0.05). Logistic regression analysis revealed Sokal score (HR=0.69, 95%CI：0.22-1.37, P＜0.05) and 3M-EMR (HR=0.47, 95%CI：0.28-0.84, P＜0.01) independently related with MR4.5, The combination of Sokal score, especially the low risk with 3M-EMR much more can predict MR4.5 (HR=0.42, 95%CI=0.21-0.82, P＜0.01). CONCLUSION There is a remarkable clinical efficacy of imatinib mesylate on CML-CP patients, moreover, low risk group has a better therapeutic response. Both Sokal score and ELTS score evaluate 3M-EMR better, both EUTOS score and ELTS score evaluate 12M-MMR better. The combination of low risk in Sokal score with 3M-EMR much more can predict MR4.5. The results of this study provide the reference basis for evaluating the clinical therapentic efficacy and timely modifying the therapeutic regimens for CML patients, also possess the reference value for predicting the MR4.5.
Antineoplastic Agents ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Prognosis ; Risk Factors ; Treatment Outcome

OBJECTIVE: To investigate the apoptosis-inducing effect of Ginsenoside (Rh2) on human acute T lymphoblastic leukemia Jurkat cells and it mechamism. METHODS: The effects of different concentration of Rh2 (0, 10 , 20, 40 and 80 µg/ml) on the proliferation activity of Jurkat cells were detected by methyl thiazolyl tetrazolium (MTT) method, and the semi-inhibitory concentration (IC) of Rh2 on Jurkat cells at 48 h was calculated. Microscopy and Hoechst 33258 fluorescence staining were used to observe the apoptosis of Jurkat cells treated with IC Rh2 for 48 h. And then, the cell experiment was divided into 4 groups: control, Rh2 (IC), PI3K inhibitor LY294002 (50 µmol/l) and Rh2 (IC) + LY294002 (50 µmol/l). After synchronous culture for 48 h, the apoptosis and cycle changes of Jurkat cells were detected by using PI single staining and Annexin V-FITC/PI double staining, respectively. Western blot was used to detect the expression level of apoptosis-related protein BAX, BCL-2, Cleaved-Caspasase 3, cell cycle-related protein Cyclin D1 and PI3K/AKT signaling pathway-related protein AKT and p-AKT. RESULTS: Rh2 (10-80 µg/ml) inhibited the Jurkat cell proliferation in a dose-time dependent manner (r = 0.999, P＜0.01; r = 0.991; P＞0.05), accompanied by obvious morphological changes of apoptosis cells. Flow cytometry showed that compared with the control group, the cell apoptosis rate in Rh2 or LY294002 group significantly increased, and the cell cycle was mostly blocked in G0/G1 phase. However, the cell apoptosis and cell cycle block in Rh2+LY294002 group were more significant than that in Rh2 and LY294002 group. Western blot showed that compared with the control group, Rh2 significantly promoted the expression of BAX and Cleaved-Caspasase 3, inhibited the expression of BCL-2, Cyclin D1 and p-AKT, furthermore LY294002 significantly promoted this effect. CONCLUSION Rh2 can induce the apoptosis of Jurkat cells in time-dose dependent manner, moreover, Rh2 also can result in an obvious block of Jurkat cells at G0/G1, that may be closely related to a series of apoptotic signaling cascades mediated by Rh2 inhibiting PI3K/AKT pathway.
Apoptosis ; Cell Proliferation ; Ginsenosides ; Humans ; Jurkat Cells ; Phosphatidylinositol 3-Kinases ; Precursor Cell Lymphoblastic Leukemia-Lymphoma