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A successful conversion of lignocellulosic biomass into biofuel could be achieved through an ionic liquids (ILs)-pretreatment and an enzyme saccharification. In this study, the fungal strains with high cellulase productions were isolated and identified as Penicillium janthinellum FHH1 and P. oxalicum FLY4. A high cellulase production was obtained at pH 4.0 and 30 °C for P. janthinellum FHH1 with corn stalk and beef extract for 9 days and for P. janthinellum FHH1 with corn stalk and peptone for 7 days. A pH range between 4.0 to 6.0 and a temperature range from 55 °C to 60 °C were applicable for cellulase with high activities during hydrolysis. A high NaCl-stability and a relatively high ILs-stability were found for cellulase obtained from P. oxalicum FLY4. High yields of reducing sugar were obtained by enzymatic hydrolysis of the original corn stalk and ILs-pretreated corn stalk. P. oxalicum FLY4 has a great potential for ILs-assisted fungi in the degradation of lignocellulosic biomass into value-added products.

To assess the effects between MPL and JAK2V617F on the thrombosis risk and peripheral blood cell counts in patients with essential thrombocythemia (ET), we identified eligible studies from PubMed, Embase, and the Cochrane Library. Seven studies were ultimately included in this meta-analysis. All studies reported the peripheral blood cell counts of ET patients, and three of them reported the eligible thrombotic events. In comparing the effect of MPL versus JAK2V617F on thrombosis, 1257 ET patients (73 MPL + and 1184 JAK2V617F +) were included. MPL-positive (MPL +) ET patients had a higher risk of thrombosis than JAK2V617F-positive (JAK2V617F +) ET patients [RR = 1.80 (1.08–3.01), P = 0.025]. And 3453 ET patients (138 MPL + and 3315 JAK2V617F +) were included in the comparison of peripheral blood cell counts. Platelet counts of MPL + ET patients were higher than that of JAK2V617F + ET patients [WMD = 81.18 (31.77–130.60), P = 0.001]. MPL + ET patients had lower hemoglobin [WMD = − 11.66 (− 14.32 to − 9.00), P = 0.000] and white blood cell counts [WMD = − 1.01 (− 1.47 to − 0.56), P = 0.000] than JAK2V617F + ET patients. These findings indicate that the MPL mutation is a high-risk factor for thrombosis in ET patients, and it may be rational to include MPL mutation in the revised IPSET as a criterion for thrombosis prediction scores. And given the differences in peripheral blood, it is necessary to further study whether MPL + ET patients differ from JAK2V617F + ET patients in bleeding and survival.

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.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.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.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.基于非标记定量蛋白组学对原发性血小板增多症患者血小板活化相关差异蛋白的分析.基于蛋白组学技术分析原发性血小板增多症（ET）特异性蛋白标志物，寻找与血小板活化相关特异性蛋白，并进行验证.应用非标记定量蛋白组学（LFP）检测ET患者和健康者血清蛋白质谱，选取疾病组蛋白丰浓度较对照组差异倍数上调或下调1.3倍及两组蛋白丰浓度经t检验P＜0.05的蛋白定义为差异蛋白。应用KEGG和GO分析软件对差异蛋白进行生物信息学分析。组间差异蛋白平均丰浓度的差异分析采用t检验，P＜005为差异有统计学意义。选取差异蛋白，应用平行反应监测进行验证.本研究发现差异蛋白共140个，其中72个蛋白表达上调，68个蛋白表达下调。KEGG富集显示差异蛋白表达与血小板活化通路相关，与血小板活化相关的差异蛋白是GPV、COL1A2、GP1bα、COL1A1和GPVI，其中GPV、GP1bα和GPVI表达上调，COL1A2和COL1A1表达下调。对COL1A1、GP1bα、GPVI和GPV进行PRM验证，其结果与LFP蛋白组学结果一致.ET患者差异蛋白与血小板活化通路激活相关，GPV、GPVI、COL1A1和GP1bα等差异蛋白可作为ET血小板活化相关的新靶点.

Indole-3-acetic acid (IAA) plays an important role in the growth and development of plants. In this study, a series of predominant strains were isolated and identified as Enterobacter sp. with remarkable IAA-producing capabilities. The IAA-producing strains are mainly tryptophan-dependent and have significantly high yields of IAA (3477 μg mL−1 and 3378 μg mL−1). The ipdC gene encoding indole-3-pyruvate decarboxylase was identified by genomic analysis and RT-qPCR analysis, indicating the involvement of the indole-3-pyruvic acid (IPyA) pathway of IAA biosynthesis. The IPyA pathway was also confirmed by the intermediate assay. The IAA product of microbial metabolites was isolated, purified and characterized. These microbes exhibiting IAA production significantly promoted the growth of maize, increasing root length, plant height, fresh weight and dry weight. Thus, Enterobacter sp. with high IAA production has great prospects in agricultural and industrial applications.

Objective Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear. Methods A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients’ medical records. Multivariable logistic regression models were used to estimate the odds ratio (OR) and 95% confidence interval (95% CI) of the associations between comorbidities (cardiometabolic or non-cardiometabolic diseases), clinical severity, and treatment outcomes of COVID-19. Results Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30–6.32), type 2 diabetes (T2DM) (3.57, 2.32–5.49), cardiovascular disease (CVD) (3.78, 1.81–7.89), fatty liver disease (7.53, 1.96–28.96), hyperlipidemia (2.15, 1.26–3.67), other lung diseases (6.00, 3.01–11.96), and electrolyte imbalance (10.40, 3.00–26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89–12.75), CVD (8.47, 6.03–11.89), and electrolyte imbalance (19.44, 11.47–32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25–9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46–29.64) within two weeks. Conclusion Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.

BACKGROUND X-linked agammaglobulinemia is a primary immunodeficiency disease caused by gene mutations of Bruton’s tyrosine kinase (BTK). We found a new mutation point and summarized the correlation analysis and performed a literature review. CASE SUMMARY The proband was a 5-year-old boy. He was admitted to our hospital due to a recurrent cough and a fever that had persisted for a month. He had a history of multiple respiratory infections and sinusitis. There was no immunodeficiency or recurrent infection history among his family members. Agammaglobulinemia was characterized as follows: Immunoglobulin (Ig) A, 90.0 mg/dL (90-450 mg/dL); IgG, 20.0 mg/dL (800-1800 mg/dL); and IgM, 18.0 mg/dL (60-280 mg/dL). Notably, the assessment of IgG subtypes revealed the following very low levels: Subtype 1, 0.26 g/L (3.62-12.28 g/L); subtype 2, 0.10 g/L (0.57-2.9 g/L); subtype 3, 0.009 g/L (0.129-0.789 g/L); and subtype 4, 0.003 g/L (0.013-1.446 g/L). Cellular immunological test results were as follows: CD3, 74.6% (50%-84.0%); CD4, 47.3% (27.0%-51.0%); and CD8, 24.9% (15.0%-44.0%). A de novo hemizygous deletion in BTK was detected: c.902_c.904delAAG/p.E301del. Transcript levels of the mutant BTK were similar to those of the wild-type gene, though overexpression resulted in markedly reduced levels of mutant BTK (9.49% ± 1.58%), relative to the wild-type BTK (75.8% ± 2.98%, P < 0.01). CONCLUSION This case of X-linked agammaglobulinemia was attributed to a de novo hemizygous deletion mutation in BTK (c.902_c.904delAAG/p.E301del). The mutation resulted in markedly reduced BTK protein stability in vitro.

Ionic liquids (ILs) are effective solvents for lignocellulose pretreatment. Enzymatic saccharification converts pretreated lignocelluloses into valuable products, and IL-tolerant cellulase improves the enzymatic efficiency and the reuse of ILs. In this study, a fungal strain with a relatively high cellulase production was isolated and identified as Aspergillus aculeatus G1-3. The high production of β-glucosidase (1.943 U per mL), CMCase (1.303 U per mL), and FPase (0.165 U per mL) was obtained using corn stover as the carbon source and peptone as the nitrogen source. The results were obtained at pH 8.0 and 30 °C with an inoculation size of 3% (volume per volume) for 7 days. A mutant strain Aspergillus aculeatus P6 with β-glucosidase (7.023 U per mL), CMCase (1.543 U per mL), and FPase (0.098 U per mL) was obtained by 60Co-γ irradiation. The cellulase activity was measured at pH 5.0 and 60 °C for enzymatic hydrolysis. The cellulase from mutant strains was stable in different concentrations of 1-ethyl-3-methylimidazolium acetate. Enzymatic saccharification of the original corn stover and ILs-pretreated corn stover was successfully performed with high sugar yields. The mutant strains of Aspergillus aculeatus have great potential for their further application in the conversion of lignocellulosic biomass into biofuels.

Background Previous studies have shown that DNA methylation plays a significant role in myelodysplastic syndrome (MDS). In addition to hypermethylation, aberrant hypomethylation can result in the transcriptional activation of oncogenes in cancer, including MDS. Therefore, drugs targeting DNA hypomethylation are needed for the treatment of MDS. This study aimed to investigate whether As2S2 promoted hypomethylation by increasing DNA methyltransferases (DNMTs) expression in MDS. Patients and methods Ten bone marrow samples from MDS patients and 3 healthy donors were obtained for the examination of the DNA methylation with a Human Methylation 850K Bead Chip. The mRNA expressions for the DNMTs in the ten MDS patients and 3 controls were compared by Q-PCR. Then, the MDS cell line SKM-1 was treated with As2S2. After 2 days of treatment, Human Methylation 850K Bead Chip was applied to analyze the changes of gene methylation status in the cells. Q-PCR and Western blot were taken to test the changes of mRNA and protein expressions for DNMTs in SKM-1 cells after treatment. Results Five hundred ninety-two abnormally hypomethylated genes were found in MDS patients compared to those in controls by Human Methylation 850K. The mRNA expressions of DNMTs (DNMT1, DNMT3a and DNMT3b) in MDS patients were significantly lower than those in healthy individuals. The IC50 value of As2S2 for SKM-1 cells was 4.97 μmol/L.Treatment with As2S2 at 2 μmoL/L resulted in significant alterations in the methylation levels at 1718 sites in SKM-1 cells compared to those in the controls. Hypermethylation was observed in 1625 sites (94.58%), corresponding to 975 genes, compared to those in the controls. Finally, the expression levels of DNMTs (DNMT1, DNMT3a, and DNMT3b) significantly increased in SKM-1 cells treated with As2S2 at 2 μmoL/L and 4 μmoL/L. Conclusion These data show a potential clinical application of As2S2 as an innovative hypermethylation agent in MDS.

Ruxolitinib is a targeted drug to treat myelofibrosis (MF). Ruxolitinib has significant advantages in spleen reduction and increasing 5-year overall survival (OS), and ruxolitinib-based combinations might provide more benefits than ruxolitinib monotherapy. In this review, we focus on the data of ruxolitinib-based combinations therapies and treatment-related adverse events (AEs) and safety. We analyzed and summarized the data of ruxolitinib-based combinations. Ruxolitinib combined with prednisone + thalidomide + danazol (TPD), panobinostat, pracinostat, azacytidine, or hydroxyurea has well reduced spleen. Ruxolitinib combined with danazol or TPD had well therapies in improvement of hemoglobin (Hgb) and platelets (PLT). Most ruxolitinib-based combinations therapies showed a superior benefit on reduced treatment-related AEs than ruxolitinib monotherapy. Treatment-related AEs and dose modification affect the safety and tolerability of ruxolitinib-based combinations. Genetic testing before treatment is recommended. To provide better clinical guidance, comparisons of these randomized controlled trials with the trials of ruxolitinib alone are necessary. This review suggests that the clinical application of ruxolitinib-based combinations is worth waiting for.

Phosphate-solubilizing microorganisms have been considered as a novel alternative approach to provide phosphate fertilizers that promote plant growth. In this study, three strains were isolated and identified as Penicillium oxalicum FJG21, Penicillium oxalicum FJQ5, and Bacillus subtilis BPM12, with a relatively high phosphate-solubilizing activity. Various phosphate sources were investigated, and Ca3(PO4)2 was identified as the effective phosphate source. Factors governing the phosphate-solubilizing activity of the strains included carbon and nitrogen sources, initial pH, and fermentation time. A high soluble phosphorus content was achieved with 529.0 μg·mL-1, 514.0 μg·mL-1, and 330.7 μg·mL-1 for Penicillium oxalicum FJG21, Penicillium oxalicum FJQ5, and Bacillus subtilis BPM12, respectively. An inverse correlation of the quantity of soluble phosphorus content and the pH value of the medium was observed. In addition, Bacillus subtilis BPM12 displayed a prominent capability of producing indole acetic acid. Penicillium oxalicum FJG21 and Penicillium oxalicum FJQ5 exhibited high cellulase activities. These phosphate-solubilizing microorganisms with good phosphate-solubilizing capability and growth-promoting ability are the promising strains for agricultural utilization.

To analyze the overall survival (OS) of elderly acute myeloid leukemia (AML) patients treated with oral arsenic-containing Qinghuang Powder (青黄散, QHP) or low-intensity chemotherapy (LIC). Forty-two elderly AML patients treated with intravenous or subcutaneous LIC (1 month for each course, at least 3 courses) or oral QHP (3 months for each course, at least 2 courses) were retrospectively analyzed from January 2015 to December 2017. The main endpoints of analysis were OS and 1-, 2-, 3-year OS rates of patients, respectively. And the adverse reactions induding bone marrow suppression, digestive tract discomfort and myocardia injury were observed. Out of 42 elderly AML patients, 22 received LIC treatment and 20 received QHP treatment, according to patients’ preference. There was no significant difference on OS between LIC and QHP patients (13.0 months vs. 13.5 months, >0.05). There was no significant difference on OS rates between LIC and QHP groups at 1 year (59.1% vs. 70.0%), 2 years (13.6% vs. 15%), and 3 years (4.6% vs. 5.0%, all >0.05). Furthermore, there was no significant difference of OS on prognosis stratification of performance status > 2 (12 months vs. 12 months), age> 75 year-old (12.0 months vs. 12.5 months), hematopoietic stem cell transplant comorbidity index >2 (12 months vs. 13 months), poor cytogenetics (12 months vs. 8 months), and diagnosis of secondary AML (10 months vs. 14 months) between LIC and QHP patients (>0.05). QHP may be an alternative treatment for elderly AML patients refusing LIC therapy.

To investigate the expressions of microRNAs in peripheral blood of patients with primary immune thrombocytopenia(ITP) and its correlation with the imbalance of Th1/Th2 cells.Thirty patients with ITP (ITP group) and 15 healthy people (control group) were enrolled．Real-time polymerase chain reaction (RT-PCR) was used to detect the expressions of six miRNAs (miR-107,miR-205-5p,miR-138-5p,miR-326,miR-1827,miR-185-5p) and Th1-specific transcription factor T-bet mRNA and Th2-specific transcription factor GATA-3 mRNA in the peripheral blood of the two groups. Th1 and Th2 cells were detected by flow cytometry. The expressions of Th1-cytokines TNF-α and IFN-γ and Th2-cytokines IL-4 and IL-10 were detected by AimPlex multiple immunoassays for Flow. The expression difference of miRNAs, mRNA, Th1, Th2 cells and cytokines of the two groups were compared, and the correlations of miRNAs to mRNA, Th1, Th2 cells and cytokines were analyzed in ITP group.The expressions of miRNAs（miR-107, miR-205-5p, miR-138-5p, miR-326, miR-1827, miR-185-5p）and Th2-specific transcription factor GATA-3 mRNA of the patients in ITP group were significantly decreased (P0.05) as compared with those in control, while the expressions of Th1 cells and Th1-specific transcription factor T-bet mRNA and Th1-cytokines TNF-α were significantly increased (P0.05), also for the ratios of T-bet mRNA/GATA-3 mRNA and Th1/Th2 cells were significantly increased (P0.05). The relative expressions of miR-107, miR-205-5p, miR-138-5p in ITP patients were negatively correlated with Th2 cells (r=-0.411, r=-0.593, r=-0.403,P0.05) and the relative expression of miR-1827 was negatively correlated with TNF-α (r=-0.390).The relative expressions of the six miRNAs in peripheral blood of patients with ITP are significantly decreased, which result in the increasing ratio of T-bet mRNA/GATA-3 mRNA, then lead to the imbalance of Th1/Th2.原发性免疫性血小板减少症患者外周血microRNAs的表达及与Th1/Th2失衡的关系.探讨原发性免疫性血小板减少症（ITP）患者外周血micoRNAs的表达量与Th1/Th2失衡的关系.纳入30例ITP患者(ITP组)及15例健康人(对照组)，采用实时定量 PCR(RT-PCR)法检测2组外周血中6种miRNA（miR-107、miR-205-5p、miR-138-5p、miR-326、miR-1827、miR-185-5p）和Th1特异性转录因子T-bet、Th2特异性转录因子GATA-3的表达水平，采用流式细胞术检测Th1、Th2细胞，AimPlex流式细胞术高通量技术检测Th1细胞因子TNF-α、IFN-γ以及Th2细胞因子IL-4、IL-10含量。比较2组间miRNAs、mRNA、Th1、Th2细胞及其分泌的主要细胞因子的差异,分析ITP患者miRNAs与Th1、Th2细胞及其分泌的主要细胞因子的相关性.与对照组相比，ITP组miRNA（miR-107、miR-205-5p、miR-138-5p、miR-326、miR-1827、miR-185-5p）和Th2细胞特异性转录因子GATA-3的表达水平均显著降低(P0.05)；Th1细胞、Th1细胞特异性转录因子T-bet及Th1细胞因子TNF-α表达水平均显著升高(P0.05)，T-bet/GATA-3和Th1/Th2比值均显著升高（P0.05）。ITP患者miR-107、miR-205-5p、miR-138-5p相对表达量与Th2细胞均呈负相关（P＜0.05，r=-0.411； r=-0.593； r=-0.403）；miR-1827相对表达量与TNF-α呈负相关（r=-0.390）.ITP患者外周血6种miRNAs表达水平显著降低，导致T-bet mRNA/GATA-3 mRNA比值升高，Th1/Th2失衡.

Mono-cationic ionic liquids (MILs) have been widely utilized for the treatment of lignocelluloses. However, the hydrophilic property of MILs increases the difficulty of isolating sugars and their reuse, which limits their industrial application. In the present study, a series of dicationic ionic liquids (DILs) were synthesized with totally different di-cations and di-anions. An alternating miscibility of DILs with water was observed depending on the dealing temperature. The functional DILs exhibiting hydrophobic and acidic properties were investigated for the treatment of corn stalk. High yields of sugars were achieved as 27.8% and 31.6% with DIL3 and DIL4, respectively. Cellulosic materials were obtained as 74.3% and 77.8% with DIL1 and DIL2 and 61.2% and 65.7% with DIL3 and DIL4, respectively. Enzymatic reaction was successfully performed with cellulosic materials after the DIL treatment. DILs were recycled and reused with no significant reduction in sugar yields. Hydrophobic DILs are a potentially effective approach to enable the successful decomposition and degradation of lignocellulosic biomass.

To explore the relationship between the efficacy of realgar for the treatment of myelodysplastic syndromes with multilineage dysplasia (MDS-MLD) and arsenic concentration in the peripheral blood of patients.In this prospective study, a total of 50 MDS-MLD patients were treated with traditional Chinese drugs containing realgar for 3 months in Xiyuan Hospital from March 2018 to January 2019. Routine blood examination as well as liver and kidney function were monitored before and after treatment. The concentration of arsenic in the peripheral blood was measured using an atomic fluorescence spectrometer after treatment. The correlation between clinical effect and arsenic concentration was analyzed by Spearman's method.The treatment response rate was 54%. Two patients (4% ) achieved complete remission, 50% (25 of 50) showed hematologic improvement, and 23 patients had stable disease (23% ). No disease progression was observed. Arsenic concentration in the peripheral blood ranged from 14.60 to 85.96 μg/L. Clinical efficacy was positively correlated with arsenic concentration (P0.05). The incidence of mild adverse reactions was 16%.A relatively high concentration of arsenic in the peripheral blood may improve the clinical efficacy of realgar in MDS-MLD patients.

Indole-3-acetic acid (IAA) plays an important role in the growth and development of plants. In this study, a series of predominant strains were isolated and identified as

A case of primary oral mucosal diffuse large B-cell lymphoma(DLBCL)due to long-term use of methotrexate(MTX)for the treatment of rheumatoid arthritis(RA)was admitted to the Department of Hematology,Fujian Medical University Union Hospital.We analyzed and discussed the clinical features,diagnosis and treatment,and prognosis of specific malignant lymphoma induced by MTX in this RA patient.Our purpose is to improve the awareness and knowledge of other iatrogenic immunodeficiency-associated lymphoproliferative disorders of clinicians and pathologists.This study provides a new reference for the clinical diagnosis and treatment of MTX-associated DLBCL.

To investigate the relation of blood arsenic concentration (BAC) with clinical effect and safety of arsenic-containing Qinghuang Powder (青黄散, QHP) in patients with myelodysplastic syndrome (MDS). Totally 163 patients with MDS were orally treated with QHP for 2 courses of treatment, 3 months as 1 course. The BACs of patients were detected by atomic fluorescence spectrophotometry at 1, 3, and 6 months during the treatment, and the effective rate, hematological improvement and safety in patients after treatment with QHP were analyzed. After 2 courses of treatment, the total effective rate was 89.6% (146/163), with 31.3% (51/163) of hematological improvement and 58.3% (95/163) of stable disease. The hemoglobin increased from 73.48 ± 19.30 g/L to 80.39 ± 26.56 g/L (P 0.05). Among 46 patients previously depended on blood transfusion, 28.3% (13/46) completely got rid of blood transfusion and 21.7% (10/46) reduced the volume of blood transfusion by more than 50% after treatment. The BACs were significantly increased in patients treated for 1 month with 32.17 ± 18.04 μ g/L (P 0.05). The adverse reactions of digestive tract during the treatment were mild abdominal pain and diarrhea in 14 cases (8.6%), and no patients discontinued the treatment. The BACs of patients with gastrointestinal adverse reactions were significantly lower than those without gastrointestinal adverse reactions (22.39 ± 10.38 vs. 37.89 ± 11.84, μ g/L, P

There is increasing interest in the application of ionic liquids for the pretreatment and fractionation of lignocelluloses. In this study, a series of functional acidic ionic liquids (ILs) with various heterocyclic organic cations were synthesized. Corn stalks were successfully fractionated into lignin, hemicelluloses, and cellulose when ultrasonically pretreated with ILs at 70 °C for 3 h, and subsequently treated with alkaline extraction. High yields of IL-isolated lignin (18.3% to 19.6%) and (8.3% to 14.6%) were obtained using ILs in the absence and presence of water, respectively. The yield of cellulose ranged from 40.0 to 77.0% from IL treatments, whereas the yield of hemicelluloses ranged from 1.1% to 17.3%. Enzymatic hydrolysis of the isolated cellulose residual produced 89.2% to 94.9% reducing sugar with 77.8% to 86.1% glucose, which corresponded to 80.5% to 91.4% enzymatic conversion of cellulose. Syringol and vanillin were found as the main lignin degradation products.

To investigate the damaging of human umbilical vein endothelial cells (HUVEC) induced by antiplatelet integrin β3 antibodies in vitro.The serum from 36 chronic ITP patients were collected, flow cytometry and monoclonal antibody specific immobilization of platelet antigen (MAIPA) assay were used to collect antiplatelet integrin β3 antibodies from the serum of the patients. After HUVEC were treated by ITP patient serum (PS) containing anti-integrin β3 antibodies, the cell damage was detected by Lactate dehydrogenase (LDH) assay, cell apoptosis was detected by flow cytometry, the expression of apoptosis-related gene Bax was detected by Reverse transcription-Quantitative real-time PCR (RT-qPCR), and expression of Apoptosis-related signaling pathway protein Akt and related protein Bax were detected by Western blot. HUVEC were treated by PS combined with Akt activator SC79, the cells damage were detected by LDH assay, apoptosis of the cells were detected by flow cytometry, the expression of apoptosis-related gene Bax was detected by RT-qPCR.Among 36 cases of serum from the chronic ITP patients, 5 patients' serum containing anti-integrin β3 antibodies were collected. After HUVEC was treated by PS, the viability of LDH was significant increased（P0.05）, so as for the apoptosis of the cells（P0.05）, the expression of gene and protein of Bax was increased up-regulated（P0.05）, the protein expression of pAkt was down-regulated（P0.05）. Comparing with HUVEC cultured with PS alone, the viability of LDH of HUVEC treated by PS combined with SC79 was significantly reduced（P0.05）, so as for the apoptosis of the cells（P0.05）, and gene expression of Bax was significantly decreased（P0.05）.Anti-integrin β3 serum can cause the damage and apoptosis of HUVEC through Akt signaling pathway，the apoptotic effects of anti-integrin β3 antibodies to HUVEC was effectively reversed by SC79.抗血小板整合素β3抗体诱导内皮细胞凋亡机制的研究.探讨抗血小板整合素β3抗体对人脐静脉血管内皮细胞（HUVEC）的损伤及其相关机制.收集36例慢性免疫性血小板减少症患者的血清，通过流式细胞术和单克隆抗体特异性俘获血小板抗原技术（MAIPA）筛选出含抗整合素β3抗体的患者血清。用抗整合素β3血清处理HUVEC后，乳酸脱氢酶(LDH)活性测定法检测HUVEC损伤情况，流式细胞术检测HUVEC凋亡情况，逆转录实时荧光定量PCR(RT-qPCR)检测HUVEC凋亡相关基因Bax表达情况，Western blot检测细胞凋亡相关信号通路蛋白Akt和相关蛋白Bax的表达情况。用抗整合素β3血清结合Akt激活剂SC79处理HUVEC后，LDH活性测定法检测HUVEC损伤情况，流式细胞术检测HUVEC的凋亡情况，RT-qPCR检测HUVEC凋亡相关基因Bax的表达情况.从36份患者血清中筛选出5份含抗整合素β3抗体血清，用这些血清处理HUVEC后，细胞外LDH活性增加（P0.05），细胞凋亡比例增加（P0.05），Bax蛋白和mRNA表达水平上调（P0.05），pAkt蛋白表达下调（P0.05）。抗整合素β3血清结合SC79处理HUVEC后，与单独使用抗整合素β3血清处理组相比，细胞外LDH活性显著降低（P0.05），HUVEC凋亡比例显著下降（P0.05），Bax基因表达下降（P0.05）.抗整合素β3抗体在体外能够引起HUVEC的损伤和凋亡，其作用机制可能与抑制Akt信号通路有关，且Akt激活剂SC79能够在体外抑制抗整合素β3抗体引起的HUVEC损伤和凋亡.

Traditional Chinese Medicine (TCM) is a practical medicine based on thousands of years of medical practice in China. Arsenic dispensing powder (ADP) has been used as a treatment for MDS patients with a superior efficacy on anemia at Xiyuan Hospital of China Academy of Chinese Medical Sciences. In this study, we retrospectively analyzed MDS patients that received ADP treatment in the past 9 years and confirmed that ADP improves patients’ anemia and prolongs overall survival in intermediate-risk MDS patients. Then, we used the MDS transgenic mice model and cell line to explore the drug mechanism. In normal and MDS cells, ADP does not show cellular toxicity but promotes differentiation. In mouse MDS models, we observed that ADP showed significant efficacy on promoting erythropoiesis. In the BFU-E and CFU-E assays, ADP could promote erythropoiesis not only in normal clones but also in MDS clones. Mechanistically, we found that ADP could downregulate HIF1A in MDS clones through upregulation of VHL, P53 and MDM2, which is involved in two parallel pathways to downregulate HIF1A. We also confirmed that ADP upregulates GATA factors in normal clones. Thus, our clinical and experimental studies indicate that ADP is a promising drug to promote erythropoiesis in both MDS and normal clones with a superior outcome than current regular therapies. ADP promotes erythropoiesis in myelodysplastic syndromes via downregulation of HIF1A and upregulation of GATA factors.

The karyotype is highly important for diagnosis and prognosis in myelodysplastic syndrome (MDS). The objective of the present study was to investigate the cytogenetic characteristics of patients with MDS in China. The karyotypes of 665 Chinese patients with MDS were analyzed, and it was identified that 298 cases (298/665, 44.8%) had abnormal karyotypes. Among the 298 patients with abnormal karyotypes, the 75 patients with trisomy 8 (+8) constituted the most common subset (75/298, 25.2%). The incidence of abnormal karyotypes was significantly higher in patients who were ≥51 years old compared with those

Additional sex combs like 1 (ASXL1) mutations are one of the most common molecular biological abnormalities in patients with primary myelofibrosis (PMF), and the effect of these mutations on prognosis remains controversial. Hence, we conducted a meta-analysis to assess the prognostic value and clinical characteristics of ASXL1 mutations in PMF patients. Eligible studies were systematically searched from PubMed, Embase, and the Cochrane Library. We extracted the hazard ratios (HRs) and their 95% confidence intervals (CIs) of overall survival (OS) and leukemia-free survival (LFS), the number of patients transformed to acute leukemia, and clinical characteristics to carry out a meta-analysis by fixed effect model or random effect model according to the heterogeneity between studies. A total of 4501 PMF patients from 16 cohorts of 14 studies were included in this meta-analysis. The results revealed that ASXL1 mutations might predict a shorter OS (HR = 2.30, 95% CI: 1.79–2.94, P P = 0.0003; the rate of acute leukemia transformation: OR = 2.06, 95% CI: 1.50–2.83, P

Single cell sequencing technology is different from traditional sequencing method, which is based on population cell average level. It has been widely used in many fields and made great progress in the application of malignant hematological diseases. In this review, the principle, methodology and application of single cell sequencing technology in malignant hematological diseases are summarized briefly, including the study of the pathogenesis in myelodysplastic syndrome, the mechanism of transformation into leukemia, accurate diagnosis and classification, differential diagnosis, evaluation of targeted drug therapeutic efficacy and exploration of biomarkers; specific diagnostic indicators for myeloproliferative diseases, progression of disease monitoring and epidemiological studies; moreover, the pathogenesis and drug resistance of acute myeloid leukemia (AML), which can provide reference for the diagnosis and research of malignant hematological diseases.单细胞测序技术在恶性血液病诊疗中的应用.摘要 单细胞测序技术与基于群体细胞平均水平的传统测序方法不同，已经在多个领域得到广泛应用，在恶性血液病中的应用也取得很大进展。本文对单细胞测序技术的原理、方法学以及在多种恶性血液病的应用进行综述，涉及到对骨髓增生异常综合征发病机制、转白机制、精确诊断与分型、鉴别诊断、靶向药疗效评价、生物标志物的探索；以及骨髓增生性疾病的特异性诊断指标的确立、疾病进展监测、流行病学相关研究；还有急性髓系白血病的发病机制以及耐药机制研究，为恶性血液病的诊断和研究提供参考.

Background: LongChaiJiangXue formula (LCJX) has the effect of not only clearing up excessive erythrocytes but also relieving clinical symptoms of Polycythemia vera (PV). Material/Methods: The chemical constitution of LCJX was identified from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Seven hundred and fifty-nine targets were identified and a total of 248 targets were screened out after discarding duplication and genes without any ID in databases. GeneCards database, OMIM database, and GEO database were searched for differential expression genes. The network was built by Cytoscape (3.7.2) software and the protein-protein interaction (PPI) networks of PV and LCJX were merged by https://string-db.org . Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment was processed by the R platform. The molecular docking technology was used to further analyze the intense of the assosiation of the compounds and targets. Results: 73 compounds of LCJX were chosen as the candidate active compounds. The compound-targets network contained 105 nods and 216 edges that presented the interaction of agents and targets. The PPI network of LCJX targets involved 59 nodes and 168 edges. The network showed that the key nodes were concentrated in signal transducer and activator of transcription-3(STAT-3), interleukin-6(IL-6), Janus kinase 2(JAK2), and vascular endothelial growth factor-A(VEGFA). The most enriched terms in the GO analysis in the GO biological processes(BP) were reactive oxygen species metabolic process, response to lipopolysaccharide, and response to oxidative stress. According to GO molecular functions(MF), the central nodes were generally enriched in cytokine receptor binding, cytokine activity, and heme binding. Regarding the GO cell components(CC), the terms included vesicle lumen, cytoplasmic vesicle lumen, and secretory granule lumen. In light of the KEGG enrichment analysis, the Hepatitis B, AGE-RAGE signaling pathway in diabetic complications, Kaposi sarcoma-associated herpesvirus infection, measles, Human cytomegalovirus infection, and JAK-STAT signaling pathway were significantly enriched. The molecular docking technology found that puerarin and saikosaponin A had relative stronger affinity with VEGFA, HIF-1A, JAK2 and STAT3 than other compounds in terms of the binding free energy. Conclusions: The effect of LCJX on PV is achieved through a series of complex mechanisms. Network pharmacology and molecular docking are powerful tools to reveal the effect of compound Chinese medicine on the disease.

Qinghuang Powder (, QHP), an oral arsenic, has become an effective drug in the treatment of myelodysplastic syndromes (MDS) in Xiyuan Hospital, China Academy of Chinese Medical Sciences for many years, and the action mechanism of the compound or active ingredient As2S2 of QHP has been elucidated. Considering the relatively safety, chemotherapy-free and convenient oral profile, QHP is widely used in the clinical treatment for MDS patients, especially for elderly patients. In this review, the authors document the efficacy and safety of oral arsenic-containing compound QHP in the treatment of MDS, with a special focus on the association of efficacy of QHP with the cytogenetics, prognostic risk, DNA methylation, gene mutation, blood arsenic concentration, mechanism of action of As2S2 and the countermeasures against adverse reactions of gastrointestinal tract.

Background Quota payment for specific diseases under global budget is one of the most typical modes of provider payment system reform in rural China. This study aimed to assess this reform mode from aspects of the total fee, structure of the fee and enrollees’ benefits. Methods A total of 127,491 inpatient records from 2014 to 2016 were extracted from the New Rural Cooperative Medical Scheme (NRCMS) database in Weiyuan County, Gansu Province. Total fee, actual compensation ratio, out-of-pocket ratio, constituent ratio of the treatment fee, constituent ratio of the inspection and laboratory fee, and length of stay were selected as dependent variables. Both generalized additive models (GAMs) and multiple linear regression models were used to measure the change in dependent variables along with year. Results Prior to the adjustment of the compensation type, out-of-pocket ratio and length of stay decreased, while total fee, actual compensation ratio, constituent ratio of the treatment fee, and constituent ratio of the inspection and laboratory fee increased. After the compensation type was adjusted, the mean of the total fee increased rapidly in 2015 and remained stable in 2016. The mean length of stay increased in 2015 but decreased in 2016. A comparison of inpatients suffering from diseases covered by quota payments and those suffering from general diseases revealed that total fee, out-of-pocket ratio, and length of stay decreased and actual compensation ratio increased for the former, whereas the opposite was true for the latter. Constituent ratio of the treatment fee and constituent ratio of the inspection and laboratory fee increased for both samples, except for the constituent ratio of the inspection and laboratory fee of quota payment diseases in 2016, which did not change. Conclusions Quota payment for specific diseases under global budget had obviously positive effects on cost control in Weiyuan, Gansu. Considering the limited coverage of quota payment for diseases, the long-term effect of this reform mode and its replicability awaits further evaluation.

Background Previous studies have shown that DNA methylation plays a significant role in myelodysplastic syndrome (MDS). In addition to hypermethylation, aberrant hypomethylation can result in the transcriptional activation of oncogenes in cancer, including MDS. Therefore, drugs targeting DNA hypomethylation are needed for the treatment of MDS. This study aimed to investigate whether As2S2 promoted hypomethylation by increasing DNA methyltransferases (DNMTs) expression in MDS. Patients and Methods Ten bone marrow samples from MDS patients and 3 healthy donors were obtained for the examination of the DNA methylation with a Human Methylation 850K Bead Chip. The mRNA expressions for the DNMTs in the ten MDS patients and 3 controls were compared by Q-PCR. Then, the MDS cell line SKM-1 was treated with As2S2. After 2 days of treatment, Human Methylation 850K Bead Chip was applied to analyze the changes of gene methylation status in the cells. Q-PCR and Western blot were taken to test the changes of mRNA and protein expressions for DNMTs in SKM-1 cells after treatment. Results Five hundred ninety-two abnormally hypomethylated genes were found in MDS patients compared to those in controls by Human Methylation 850K. The mRNA expressions of DNMTs (DNMT1, DNMT3a and DNMT3b) in MDS patients were significantly lower than those in healthy individuals. The IC50 value of As2S2 for SKM-1 cells was 4.97 μmol/L.Treatment with As2S2 at 2 μmoL/L resulted in significant alterations in the methylation levels at 1718 sites in SKM-1 cells compared to those in the controls. Hypermethylation was observed in 1625 sites (94.58%), corresponding to 975 genes, compared to those in the controls. Finally, the expression levels of DNMTs (DNMT1, DNMT3a, and DNMT3b) significantly increased in SKM-1 cells treated with As2S2 at 2 μmoL/L and 4 μmoL/L. Conclusion These data show a potential clinical application of As2S2 as an innovative hypermethylation agent in MDS.

To investigate the relationship between gene mutations and response to Compound Qinghuang Powder (复方青黄散, CQHP) in patients with myelodysplastic syndrome (MDS). Forty-three MDS patients were genotyped by ultra-deep targeted sequencing and the clinical data of patients were collected and the relationship between them was analyzed. Up to 41.86% of patients harbored genet mutations, in most cases with more than one mutation. The most common mutations were in SF3B1, U2AF1, ASXL1, and DNMT3A. After treatment with CQHP, about 88.00% of patients no longer required blood transfusion, or needed half of prior transfusions. CQHP is an effective treatment for patients with MDS, especially those with gene mutations in SF3B1, DNMT3A, U2AF1, and/or ASXL1.

To explore the effect of Qinghuang Powder (QHP,(青黄散)combined with Bupi Yishen Decoction (BPYS, 补脾益肾方) on myelodysplastic syndromes (MDS) patients with refractory cytopenia with multilineage dysplasia (RCMD) and determine the change of DNA methylation in MDS-RCMD patients after the treatment of Chinese medicine formula. All 308 MDS-RCMD patients were treated with QHP combined with BPYS for 2 months at least, absolute neutrophil count (ANC), hemoglobin (Hb), platelets (PLT), primitive bone marrow cells and chromosome karyotype were chosen as the main evaluation indexes to analyze the treatment effect according to criteria from the MDS International Working Group. Then 43 bone marrow samples from 15 MDS-RCMD patients and 28 healthy donors were obtained for the examination of DNA methylation. Gene Ontology (GO) and Pathway analysis were applied to analyze the methylation data. The overall MDS response rate to QHP was 61.68% (190/360) including hematologic improvement-neutrophil (HI-N) or hematologic improvement-erythroid (HI-E) or hematologic improvement-platelet (HI-P). Patients with anemia had a better response rate than patients with neutropenia or thrombocypenia (55.88% vs 31.54% or 55.88% vs. 36.9%). The DNA methylation microarray analysis disclosed that 4,257 hypermethylated genes were demethylated upon the treatment with QHP and BPYS. GO analysis and Pathway analysis showed that these demethylated genes were involved in a lot of tumor-related pathways and functions. QHP combined with BPYS could effectively treat MDS-RCMD patients through hematologic improvement (HI-N, HI-P or HI-E) and PLT and RBC transfusion independence due to the demethylation, thereby providing another choice for the treatment of patients with MDS-RCMD.