Expression of Key Enzymes in Glucose Metabolism in Chronic Mountain Sickness and Its Correlation with Phenotype / 中国实验血液学杂志

OBJECTIVE@#To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin.@*METHODS@#Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×109/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71+ erythrocytes and hemoglobin.@*RESULTS@#The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717).@*CONCLUSION@#CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.

Assessment strategies for drug permeability during drug discovery and development / 药学学报

Permeability is a key factor in the bioavailability of oral drugs. Therefore, in the early stage of drug discovery, accurate and efficient evaluation of drug permeability is essential. The parallel artificial membrane permeability assay (PAMPA) with Caco-2 cells model was used by the industry as early evaluation methods. At present, the Ussing chamber rat model is also widely used. This review summarizes the human data for the in vivo single-pass perfusion technique (Loc-I-Gut) – the gold standard, and then focuses on the basic principles, experimental operation, and efficiency of the three in vitro methods, with correlation to the effective permeability coefficient (Peff) and fractional absorbed (Fa) in man. We provide recommendations for the use of proper permeability methods at different stages in drug discovery and development.

First-line atezolizumab plus chemotherapy in treatment of extensive small cell lung cancer: a cost-effectiveness analysis from China / 中华医学杂志(英文版)

BACKGROUND@#IMpower 133 trial first confirmed the efficacy and safety of adding atezolizumab or placebo to first-line treatment with chemotherapy in patients with extensive-stage small-cell lung cancer (SCLC). While, overprice limited its broad use in clinical. The aim of this study was to evaluate the cost-effectiveness of atezolizumab plus chemotherapy in treatment of extensive SCLC as first line in China.@*METHODS@#A Markov model was established by extracting data from the IMpower 133 trial with untreated extensive SCLC patients. Utility values were obtained from published studies, and the costs were acquired from real world and literature. Additionally, sensitivity analyses based on a willingness-to-pay (WTP) threshold were performed to identify the uncertain parameters of Markov model.@*RESULTS@#Total costs of atezolizumab group were $48,129, while cost of chemotherapy alone was just $12,920 in placebo group. The quality-adjusted life-years (QALYs) in atezolizumab group was just 0.072 higher than that in placebo group (0.858 QALYs vs. 0.786 QALYs). The cost-effectiveness ratio between atezolizumab combination with chemotherapy and chemotherapy alone was $489,013/QALY in China. The net benefit of placebo group was significantly higher than atezolizumab group. One-way sensitivity analyses highlighted that utilities of the progression-free survival (PFS) and progression disease state in placebo group were the most influential parameter.@*CONCLUSIONS@#Atezolizumab combination therapy was not more cost-effective than chemotherapy alone at a WTP threshold of $25,929/QALY in China.

Significance of Bone Marrow Microvessel Density and Vascular- Related Factors in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical significance of bone marrow microvessel density(MVD) and angiogenesis related factors in multipic myeloma(MM).@*METHODS@#Twenty cases of MM and 20 cases of simple fracture were selected and enrolled in MM group and control group respectively. The clinical data and results of laboratorial tests were collected; the bone marrow MVD of patients was detected by using the modified plastic-embedded pathologic sections of bone marrow tissue and histochemistry staining, the expression levels of amgiogenesis-related factors including VEGF, TNF-α, HGF, TGF-α, TGF-β1, bFGF, Ang-Ⅰ, Ang-Ⅱ in bone marrow supernatant were detected by ELISA; the mRNA expression levels of above-mentioned cytokines in bone marrow mononuclear cells were detected by real time-PCR; the pearson correlation analysis was used to analyze the correlation of MVD with VEGF, HGF and bFGF levels.@*RESULTS@#The MVD in MM group was significantly higher than that in control group (P＜0.001); the mRNA expression of VEGF, TGF-α, TGF-β1 and HGF in bone marrow mononuclear cells of MM group was higher than that of control group(P＜0.001); the levels of VEGF, HGF, bFGF and THF-α in bone marrow supernatant of MM group were higher than those in control group(P＜0.05), moreover, the MVD positively correlated with levels of VEGF, HGF and bFGF in bone marrow(r=0.488, 0.472 and 0.457).@*CONCLUSION@#The MVD and levels vessel-related factors in bone marrow supernatant of MM patients increase, among which the levels of VEGF and HGF in bone marrow supernatant are consistant with those mRNA expression level in bone marrow mononuclear cells, moreover, the MVD possitively cerrelates with levels of VEGF, HGF and bFGF in bone marrow supernatant, suggesting that the changes of bone marrow microenvironment vassel-related factors play an important role in angiogenesis and pathogenesis of multiple myeloma.

Effect of HIF-1α on Angiogenesis-Related Factors in K562 Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the mechanisms of angiogenesis in chronic myeloid leukemia (CML) through detecting the levels of angiogenesis-related factors secreted from K562 cells after overexpression and interference of HIF-1α gene in K562 cells.@*METHODS@#The K562 cells were transfected by lentiviruses carried and interfered HIF-1α gene, then the transtected K562 cells with carried and interfered with HIF-1α gene were enrolled in overexpression and interference groups respectively, at the same time the K562 cells transfected by the empty virus were enrolled in control group. The cells were harvested after culture for 72 hours under normoxid condition. The transfection efficient in 3 groups was detected by fluorescence microscopy; the mRNA expression of HIF-1α gene and angiogenesis-related factors was detected by RT-PCR; the concentration of angiogenesis-related factors in the caltured supernatant was detected by ELISA.@*RESULTS@#The optimal MOI of K562 cells transfected with lentivirus was 10 and the transfection efficiency was about 50%. The positive rate of transfection after screening by puromycin was more than 90%. The mRNA expression of ANG-I, ANG-II, TGF-α and VEGF in the interference group was lower than that in the over-expression group, and the TGF-β1 mRNA expression in the interference group was higher than in the over-expression group. The mRNA expression of ANG-I and VEGF in the interference group was lower than that in the control group. TGF-αdid not could be detected, and the culture supernatant concentration of ANG-I and TNF-α in the interference group was lower than in the over-expression group, while the VEGF concentration in the interference group was higher than that in the over-expression group. All of the above-mentioned differences were statistically significant (P＜0.05).@*CONCLUSION@#The positive K562 cells transfected with leutivirus have been harvested by screening with puromycin. The HIF-1α mRNA positively regulates the mRNA expression of ANG-1, ANG-2, TGF-α, VEGF in K562 cells, promotes the antocrine ability of ANG-1 and TNF-α, moreover not stimulates the autocrine of TGF-α, the up-regulation of HIF-1α expression can inhibit the expression TGF-β1 in K562 cells and the autocrine of TGF-β1.

Age of diagnosis of autism spectrum disorder in children and factors influencing the age of diagnosis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the age of diagnosis of autism spectrum disorder (ASD) and the factors influencing the age of diagnosis in children.</p><p><b>METHODS</b>A retrospective analysis was performed for the clinical data of 1 691 children who visited in the Children's Hospital of Chongqing Medical University for the first time and were definitely diagnosed with ASD between February 2011 and July 2017. A multiple linear regression model was used to identify the factors influencing the age of diagnosis of ASD.</p><p><b>RESULTS</b>The ASD children had a mean age of 35±17 months (range 9-175 months) at diagnosis. Of all 1 691 children, the children who received a diagnosis of ASD at the age of 24-35 months accounted for the highest proportion (46.13%, 780/1 691), followed by those at the age of ≥36 months (33.41%, 565/1 691). The multiple linear regression analysis showed that the children who had language disorders or lived in the main urban area or whose parents had a high education level had a younger age at diagnosis than other children (P<0.05).</p><p><b>CONCLUSIONS</b>Most ASD children have an age of 24-35 months at diagnosis. The age of diagnosis of ASD is associated with children's symptoms, living area, and parents' education level.</p>

Changes of Neutrophil CD64 in Patients with Hematological Mali-gnancies Combined with Bacterial Infections / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the changes of CD64 on surface of neutrophils in patients with hematological malignancies combined with bacterial infections.</p><p><b>METHODS</b>Ninety-seven patients with hematological tumor admitted in our hospital from August 2014 to February 2016 were selected and divided into 2 groups: infection group(50 cases) and noninfection group(47 cases) according to their symptoms, physical sings and blood culture results for microbiologic detection. The CD46 index on surface of neutrophils, serum C- reactive protein (CRP), neutrophil count (NC) and procalcitonin (PCT) level were detected by flow cytometry. After treatment of infection patients, the CD46 index, CRP, PCT and NC were detected again.</p><p><b>RESULTS</b>Before antibacterial treatment of patients, the CD64 index in infection group was significantly higher than that in noninfection group, the CRP, PCT and NC levels in infection group also were higher than those in noninfection group; after antibacterial treatment, the CD64 index, CRP, PCT and NC levels in infection group decresed.</p><p><b>CONCLUSION</b>The CD64 index in hematologic malignancy patients complicated with bacterial infections significantly increased, after antibacterial treatment these indexes decreases, confirming the value of CD64 in the diagnosis of bacterial infections for patients with hematologic malignancies.</p>

Epigenetic mechanisms of chronic pathological pain: Research progress / 第二军医大学学报

Pathological pain refers to the pain that caused by the tissue injury due to trauma, infection, tumor, etc. It may turn into chronic pain if it lasts for more than one month or remains when the tissue injury is healed. Patients with chronic pain often suffer insomnia. anxiety, depression and other mental disorders. which may seriously impair their physical function and quality of life, subsequently leading to many social and economic problems. Recent studies have found that epigenetic regulation may explain, at molecular levei, the pathogenesis of pathological pain, including inflammation pain, neuropathic pain and psychological pain, which may lead to the devclopment of its thcrapcutic means. In this article, we reviewed the latest research progress of epigenetic regulation of chronic pathological pain. including DNA methylation, histone acetylation and miRNA activity.

Significance of Plasma HMGB1, IFN-γ, IL-4 and TLR4 Expression on CD4T Cell Surface in Patients with Immune Thrombocytopenia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of plasma HMGB1, IFN-γ, IL-4 and CD4T cell surface TLR4 expression on the immune thrombocytopenia(ITP).</p><p><b>METHODS</b>Twenty-five patients diagnosed as ITP in our hospital from October 2014 to October 2015, and 20 healthy persons as controls were selected. The ELISA was used to detect the plasma levels of HMGB1, IFN-γ and IL-4 in 2 groups; the flow cytometry was used to detect the expression level of TLR4 on the surface of CD4cells. The relatienship between different parameters was analyzed.</p><p><b>RESULTS</b>Before treatment, the plasma HMGB1 level in ITP group was significantly higher than that in control group (t=4.259, P<0.01), while after treatment it significantly decreased and close to level in control group (t=1.267, P>0.05). The plasma IFN-γ level detected in ITP group before and after treatment was not significantly different from level in control group (P>0.05), while the IL-4 level in ITP group before treatment was significantly lower than that in control grup (t=2.107, P<0.05), but the IL-4 level in ITP group after treatment was significantly higher than that in control group (t=2.107, P<0.05). The plasma IFN-γ/IL-4 ratio in ITP group before treatment was significantly higher than that in control group (t=5.436, P<0.01), but it obviously decreased and was slightly lower than that in control group after treatment. The expression level of TLR4 in ITP group before and after treatment was all significantly lower than that in control group (P<0.01). The HMGB1 level in ITP group was directly proportional to the CD3content (r=0.824, P<0.01), however it not significantly related with CD4content (r=0.074, P>0.05), but than the HMGB1 level in ITP group was directly proportional to CD8content (r=0.844, P<0.01) and to IL-4 content (r=0.784, P<0.01), and was inversely proportional to IFN-γ level(r=-0.814,P<0.01),and also was inversely proportional to IFN-γ /IL-4 ratio(r=-0.887,P<0.01),and directly proportional to TLR4 expression level(r=0.772,P<0.01).</p><p><b>CONCLUSION</b>The HMGB1 and TLR4 expression levels in ITP patients are higher, and clinical therapy can relieve the disease by targetly control in HMGB1 and TLR4 expression.</p>

Role of CD8(+)T cells and their secreted cytokines in the pathogenesis of aplastic anemia / 中国实验血液学杂志

Aplastic anemia(AA) is mostly considered as an immune-mediated bone marrow failure syndrome, characterized by pancytopenia and bone marrow hypoplasia. The pathogenesis of AA is complicated, until now it is not fully understood. Further study on the pathological mechanism will be helpful for the diagnosis and treatment of AA. CD8(+) T cells and their secreted cytokines play important roles in the abnormal immunity during the process of AA. Thus, this review focuses on the role of CD8(+) T cells and their secreted cytokines in the pathogenesis of AA.

Expression of IL-27 in multiple myeloma and its cell lines / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the expression and significance of IL-27 in patients with multiple myeloma (MM) and in the supernatant of MM cell lines U266 and RPMI8226 cells culture medium.</p><p><b>METHODS</b>A total of 20 MM patients and 20 controls were enrolled in this study. The expressions of IL-27 and IL-6 in MM patient's blood plasma, and the expression of IL-27 in U266 and RPMI8226 culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-27 in mononuclear cells of MM patients' peripheral blood was measured by real-time quantitative polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The levels of IL-27 in plasma of MM patients and normal controls were (61.82 ± 8.01) ng/L and (8.29 ± 4.41) ng/L (P<0.05), and those of IL-6 were (45.62 ± 1.24) ng/L and (2.27 ± 0.18) ng/L (P<0.05), respectively. The levels of IL-27 in U266 and RPMI8226 culture supernatant were (50.06 ± 5.72) ng/L and (335.47 ± 41.88) ng/L. RT-PCR revealed that the levels of IL-27 mRNA were up-regulated in MM patients compared to controls.</p><p><b>CONCLUSION</b>High expression of IL-27 is observed in MM patients and MM cell lines U266 and RPMI8226 cells. IL-27 may play a important role in pathogenesis of MM.</p>