OBJECTIVE:To investigate the relationship between mutated genes and clinical features in patients with essential thrombocythemia (ET).METHODS:The clinical data of 69 patients with ET from October 2018 to March 2022 were retrospectively analyzed. According to driver mutation type, patients were divided into JAK2 group, CALR group and triple-negative group. The sex, age, cardiovascular risk factors, thrombosis, splenomegaly, routine blood test and coagulation status of patients in three groups were analyzed.RESULTS:Among 69 ET patients, 46 cases were associated with JAK2 mutation, 14 cases with CALR mutation, 8 cases with triple-negative mutation, and one with MPL gene mutation. There were no significant differences in age and sex among the three groups (P >0.05). The highest thrombotic rate was 26.09% (12/46) in JAK2 group, then 12.5% (1/8) in triple-negative group, while no thrombotic events occurred in CALR group. The incidence of splenomegaly was the highest in JAK2 group (34.78%), while no splenomegaly occurred in triple-negative group. The white blood cell (WBC) count in JAK2 group was (9.00±4.86)×109/L, which was significantly higher than (6.03±2.32)×109/L in CALR group (P <0.05). The hemoglobin (Hb) and hematocrit (HCT) in JAK2 group were (148.42±18.79) g/L and (0.44±0.06)%, respectively, which were both significantly higher than (131.00±15.17) g/L and (0.39±0.05)% in triple-negative group (P <0.05). The platelet (PLT) in JAK2 group was (584.17±175.77)×109/L, which was significantly lower than (703.07±225.60)×109/L in CALR group (P <0.05). The fibrinogen (Fg) in JAK2 and triple-negative group were (2.64±0.69) g/L and (3.05±0.77) g/L, respectively, which were both significantly higher than (2.24±0.47) g/L in CALR group (P <0.05, P <0.01). The activated partial thromboplastin time (APTT) in triple-negative group was (28.61±1.99) s, which was significantly decreased compared with (31.45±3.35) s in CALR group (P <0.05).CONCLUSIONS:There are differences in blood cell count and coagulation status among ET patients with different driver gene mutations. Among ET patients, JAK2 mutation is most common. Compared with CALR group, the thrombotic rate, WBC and Fg significantly increase in JAK2 group, while PLT decrease. Compared with triple-negative group, the incidence of splenomegaly and HCT significantly increase. Compared with CALR group, Fg significantly increases but APTT decreases in triple-negative group.

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.

ObjectiveTo further understand the factors that promote thrombosis in essential thrombocythemia （ET） that were not included in IPSET-thrombosis （International Prognostic Score of Thrombosis for ET）. MethodsA systematic evaluation was conducted on 292 studies by retrieving PubMed，Web of Science, and The Cochrane Library. The incidence and relative risk ratio （RR） of thrombosis in ET patients with different risk factors were calculated. The confidence levels for the outcome were analyzed in combination with RR values and recommendation scores (RS) to find the thrombotic risk factors in ET patients. ResultsOut of 9 risk factors that promoted ET thrombosis, there were 4 factors in IPSET-thrombosis, including history of thrombosis （35.0%, RR=3.30, 95% CI : 2.30-4.75）, JAK2V617F positive （21.0%, RR=2.28, 95%CI : 1.14-4.56）, age over 60 years old （ 22.4% , RR=1.95, 95%CI : 1.17-3.23） and cardiovascular risk factors （17.7%, RR =1.90, 95%CI : 1.08-3.35）. The remaining five risk factors, not listed in IPSET-thrombosis, included monoclonal type of ET based on X-chromosome inactivation pattern （XCIP, 30.6%, RR=4.49, 95% CI: 1.58-12.77）, high JAK2V617F burden （33.9%, RR=3.69, 95%CI: 1.70-7.99）, leukocytosis （18.7%, RR=3.05, 95%CI: 1.81-5.13）, MPL mutation （22.1%, RR=2.40, 95%CI: 1.08-5.33） and splenomegaly （25.6%, RR=1.76, 95%CI：1.29-2.40）. ConclusionAmong the risk factors promoting thrombosis in ET patients, five thrombotic risk factors not included in the IPSET-thrombosis have been identified.

Abnormal platelet activation can lead to thrombosis in essential thrombocythemia (ET) and thus impact patient prognosis. Platelet activation-associated proteins are key molecules for platelet activation. However, it is unclear which proteins are most closely associated with the disease's prognosis. To determine which platelet activation-related proteins can be employed as ET patient prognosis predictors, we used label-free quantification (LFQ) and parallel reaction monitoring (PRM) technology and first determined the serum proteomic expression levels and the differential proteins of ET patients. Then, based on the IPSET (International Prognostic Score for ET), the differential protein associated with the prognostic score was found. To investigate potential processes affecting prognosis, the connection of this protein with prognostic markers, such as thrombotic history, age, white blood cell count, coagulation factors, and inflammatory factors, were further examined. The levels of platelet activation-related proteins GPIbα, SELP, PF4, MMP1, and FLNA were significantly higher in ET patients, according to LFQ and PRM analyses (p < 0.01). Based on regression analysis of the IPSET prognostic score, it is suggested that the SELP level was positively correlated with the prognostic score and prognostic risk factor analysis (p < 0.05). Further regression analysis of SELP with coagulation factors showed that antithrombin (AT-III) was negatively correlated with SELP levels (p < 0.05). Further regression analysis of the inflammatory factors with AT-III and SELP revealed that IL-10, IL-12P70, and IL-31 were negatively correlated with AT-III and SELP (p < 0.01). Platelet activation pathway-related proteins are expressed more frequently in ET patients, and serum SELP may be a prognostic marker for these individuals by encouraging leukocyte increase and inflammatory factor expression and causing aberrant coagulation.

Thrombosis is a major cause of morbimortality in patients with polycythemia vera (PV). Furthermore, neutrophils play a significant role in thrombosis, but their role in the pathogenetic mechanisms of PV is not well characterized. Therefore, we investigated the role and mechanisms by which neutrophils regulate thrombosis in PV patients. Univariate and multivariate logistic regression analysis of clinicopathological factors was performed to determine the independent risk factors of thrombosis in PV. Pearson's correlation analysis was performed to determine the relationship between absolute neutrophil count (ANC) and the hypercoagulable state in PV patients. Bioinformatics analysis of the GSE54644 dataset was used to identify hemostasis-related pathways in neutrophils of PV patients. Weighted gene co-expression network analysis (WGCNA) of the integrated dataset (GSE57793, GSE26049 and GSE61629) was used to identify neutrophils-related genes and pathways associated with thrombosis in PV. Ingenuity pathway analysis (IPA) was performed to identify the differentially activated pathways in PV patients with or without thrombosis using GSE47018 dataset. Our data showed increased ANC in PV patients. Multivariate logistic regression analysis showed that ANC was an independent risk factor for the thrombotic events in PV patients before or at diagnosis. ANC correlated with the hypercoagulable state in PV patients. Neutrophil extracellular traps (NETs) pathway was significantly enriched in the neutrophils of PV patients. IPA results demonstrated that PRKCD-mediated NETs pathway was hyperactivated in PV patients with thrombosis. In summary, ANC was an independent risk factor for the thrombotic events in PV patients before or at diagnosis, and PRKCD-mediated NETs pathway was aberrantly activated in the neutrophils of PV patients and was associated with the thrombotic events.

目的 论证柴丹泻血方治疗肝郁血瘀证原发性血小板增多症(essential thrombocythemia,ET)的科学性.方法 基于ET的发病机制以及治疗现状,分析中医药治疗ET的优势;基于其中医病因病机,分析柴丹泻血方的组方用药特点;基于柴丹泻血方组方药物及其活性成分的现代药理学研究,探讨柴丹泻血方治疗肝郁血瘀证ET的机制;基于柴丹泻血方的网络药理学研究,揭示柴丹泻血方治疗肝郁血瘀证ET的关键通路与靶点.结果 ET的发病及血栓形成与JAK2V617F突变及JAK-STAT通路活化关系密切,但现有治疗方法存在不良反应不能耐受及药物抵抗等问题,部分中药有降血小板和抗凝的作用,但尚无特效中药;柴丹泻血方基于该病肝郁血瘀的核心病机,具有疏肝化瘀的功效;现代药理学研究表明柴丹泻血方组方及其活性成分具有抑制JAK-STAT通路和炎性因子、抗血小板聚集与抗凝血的作用;网络药理学研究显示该方可以抑制JAK-STAT信号通路,具有抑制巨核系增殖和防治血栓形成的作用.结论 柴丹泻血方疏肝化瘀,组方科学,成分清楚,机制明确,治疗肝郁血瘀证ET具有科学性.

Background Thrombosis is a common complication of myeloproliferative neoplasm (MPN) and its major cause of disability and death. With the development of next-generation gene sequencing technology, the relationship between non-driver mutations and thrombotic risk factors has also attracted close attention. Methods The clinical data of 125 MPN patients (75 ET and 50 PV) attending Xiyuan Hospital of the China Academy of Chinese Medical Sciences between January 2018 and March 2022 were retrospectively analyzed. A multivariate analysis of risk factors for thrombosis was performed using a Cox proportional hazards model. Results A total of 125 patients with MPN (75 ET and 50 PV) were evaluated, with a median age at diagnosis of 50 (18-78) years, 67 (53.6%) males and 58 (46.4%) females. The mutation rates of driver genes JAK2V617F, CALR, and MPL were 70.4%, 12.8%, and 0.8%, respectively. At least one non-driver mutation was detected in 36% of patients, with the highest rate of mutations in TET2 (12%), followed by ASXL1 (9.6%). Thrombotic events occurred in 35 of 125 patients (28.0%), and the incidence of thrombotic events was 21.3% (16/75) in ET patients and 38.0% (19/50) in PV patients, respectively. For 125 MPN patients, univariate analysis revealed significant associations between thrombosis and age at diagnosis, age >60 years at diagnosis, JAK2V617F mutation, TET2 mutation, WBC, NLR, cardiovascular risk factors and history of remote thrombosis. In the multivariate analysis that included the above 8 factors as covariates, age at diagnosis (HR: 1.03, 95％ CI: 1.00-1.06; P = 0.046), TET2 mutation (HR: 2.84, 95％ CI: 1.26-6.36; P = 0.011), NLR (HR: 1.18, 95％ CI: 1.06-1.32; P = 0.003), and history of remote thrombosis (HR: 5.33, 95％ CI: 1.93-14.70; P = 0.001) remained risk factors for thrombosis in MPN patients. In the subgroup analysis of ET and PV patients, univariate analysis was performed for the parameters, and factors with differences were used as covariates for further multivariate analysis. In ET patients, multivariate analysis showed that TET2 mutation and a history of remote thrombosis were independent risk factors for thrombosis in ET patients, with an HR of 4.1 (95% CI: 1.40-12.01; P = 0.01) for TET2 mutation and 6.89 (95% CI: 1.45-32.68; P = 0.015) for history of remote thrombosis (Table 1). In PV patients, multivariate analysis observed neutrophil-to-lymphocyte ratio (NLR) (HR: 4.77, 95% CI: 1.33-17.16; P = 0.017) and a history of remote thrombosis (HR: 1.67, 95% CI: 1.03-1.32; P = 0.014) as independent risk factors for thrombosis, however, no association of TET2 mutation with thrombosis was observed in PV (Table 1). Among 125 patients, TET2 mutations were detected in 15 patients (9 ET and 6 PV), with a mutation rate of 12%. Among them, 11 patients combined with JAK2V617F mutation (6 ET and 5 PV), 2 ET patients combined with CALR mutation and 1 ET patient combined with ASXL1 mutation. Two loci mutations of TET2 were detected in one patient with PV. The details of the 15 MPN patients with TET2 mutation are shown in Table 2. Our study above have shown that TET2 mutation is an independent risk factor for thrombosis in ET patients, and to understand their clinical characteristics and coagulation status, we compared the clinical characteristics and coagulation function of TET2 mutated and unmutated patients. We found that the age of ET patients with TET2 mutation was significantly higher than that of patients without TET2 mutation (P < 0.05) . In terms of coagulation function, compared with ET patients without TET2 mutation, patients with TET2 mutation were in a relatively hypercoagulable state, as evidenced by a significant increase in D-Dimer (P < 0.01) and a significant decrease in antithrombin Ⅲ (AT-III) (P < 0.05). Conclusion TET2 mutation is an independent risk factor for thrombosis in ET patients, but no such association was observed in PV. Therefore, TET2 mutation is more valuable in predicting thrombosis in ET patients compared to PV. ET patients with TET2 mutation were older and in a relatively hypercoagulable state of coagulation compared to TET2 unmutated patients. However, prospective studies with more patients are needed to validate our results and to fully elucidate the association. Our study reports risk factors for thrombosis in MPN patients in Asia and complements the data on the relationship between non-driver mutations and thrombotic events in Asian MPN patients, providing a reference for future larger studies. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Thrombosis is a common complication of myeloproliferative neoplasm (MPN), and it is a major cause of disability and death. With the development of next-generation gene-sequencing technology, the relationship between non-driver mutations and thrombotic risk factors has also attracted considerable attention. To analyze the risk factors of thrombosis in patients with essential thrombocythemia (ET) and polycythemia vera (PV), we retrospectively analyzed the clinical data of 125 MPN patients (75 ET and 50 PV) and performed a multivariate analysis of the risk factors of thrombosis using a Cox proportional risk model. Among the 125 patients, 35 (28.0%) had thrombotic events, and the incidence of thrombotic events was 21.3% and 38.0% in ET and PV patients, respectively. In ET patients, the multivariate analysis showed that a TET2 mutation and history of remote thrombosis were independent risk factors for thrombosis in ET patients, with an HR of 4.1 (95% CI: 1.40–12.01; p = 0.01) for TET2 mutation and 6.89 (95% CI: 1.45–32.68; p = 0.015) for a history of remote thrombosis. In PV patients, the multivariate analysis presented the neutrophil-to-lymphocyte ratio (NLR) (HR: 4.77, 95% CI: 1.33–17.16; p = 0.017) and a history of remote thrombosis (HR: 1.67, 95% CI: 1.03–1.32; p = 0.014) as independent risk factors for thrombosis, with no significant change in the risk of thrombosis in patients with TET2 mutations. A further analysis of the clinical characteristics and coagulation occurring in ET patients with a TET2 mutation revealed that the values of age and D-dimer were significantly higher and antithrombin III was significantly lower in TET2-mutated ET patients compared to TET2-unmutated patients. In summary, TET2 mutation may be more valuable in predicting thrombosis in ET patients than in PV patients. ET patients with a TET2 mutation are older and present differences in coagulation compared to TET2-unmutated patients.

AbstractObjective: To analyze the DNA methylation gene mutations of myeloproliferative neoplasm (MPN), and preliminarily explore its clinical features. METHODS:Next-generation sequencing technology was used to detect 31 MPN-related genes in 105 cases of MPN patients ［40 cases of polycythaemia vera (PV), 65 cases of essential thrombocythemia (ET)］, and to analyze the relationship between DNA methylation gene mutations and clinical features. RESULTS:15 mutation types were detected in 105 patients (88 mutations in total), and the total mutation detection rate was 87.6% (92/105). A total of 23 mutations in 4 DNA methylation genes (TET2, DNMT3A, IDH1, IDH2) were detected in 22 patients. The mutation rate of DNA methylation genes was 21.0%, mainly in the form of double mutations, including JAK2 V617F and TET2 (n=10), JAK2 V617F and DNMT3A (n=4), CALR and TET2 (n=2), JAK2 V617F and IDH1 (n=1). Compared with MPN patients without DNA methylation gene mutations, the proportion of women with DNA methylation gene mutations and the white blood cell count (WBC) were significantly higher (P<0.05). Compared with MPN patients with triple-negative driver genes, the proportion of women with DNA methylation gene mutations, age, WBC, platelet count (PLT), and neutrophil-to-lymphocyte ratio (NLR) were significantly higher (P<0.05). The remaining difference was not statistically significant (P>0.05). The MPN10 score, the incidence of thrombotic events, and the proportion of medium-risk and high-risk patients with DNA methylation gene mutations were significantly higher than those of MPN patients without DNA methylation gene mutations (P<0.05). CONCLUSION:The mutation rate of DNA methylation genes was 21.0%, mainly coexisting in the form of double mutations. The proportion of women with DNA methylation gene mutations in MPN patients and WBC is high, the symptom load is heavy, the incidence of thrombosis is high, and the proportion of medium-high-risk patients is high, suggesting that their prognosis may be poor.

We explored the mechanisms and molecular targets of Ejiao Siwu Decoction (EJSW) for treating primary immune thrombocytopenia (ITP) using network pharmacology and molecular docking. Active compounds of EJSW were identified by high-performance liquid chromatography-diode array detector (HPLC-DAD) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) and their targets were obtained from HERB and SwissTargetPrediction, and ITP targets were obtained from Comparative Toxicogenomics Database (CTD) and GeneCards. STRING and Cytoscape were used for protein-protein interaction (PPI) network analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses by WebGestalt yielded a gene-pathway network, Autodock molecular docking was applied to screen targets and active compounds, and cytokines were detected using a cytometric bead array (CBA) human inflammation kit. We identified 14 compounds and 129 targets, and 1,726 ITP targets. RAC-alpha serine/threonine-protein kinase (AKT1), tumour necrosis factor (TNF), interleukin-6 (IL6), caspase-3 (CASP3) and tumour suppressor protein (TP53) were core targets (nodes and edges). Functional annotation identified cofactor binding and coenzyme binding, and 20 significantly enriched pathways. Active compounds of EJSW were successfully docked with ITP targets. Tumour necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) were upregulated in ITP patients, vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor D (VEGF-D) were downregulated, and EJSW treatment reversed these trends. EJSW may regulate key ITP targets based on the in silico analyses, and protect vascular integrity through AGE-RAGE signalling, complement and coagulation cascades, and VEGF signalling by downregulating TNF-α, IL-1β and other inflammatory factors.

Polycythemia vera (PV) is a malignant clonal hematological disease of hematopoietic stem cells characterized by the proliferation of peripheral blood cells, and JAK2 mutation is one of the main causes of PV peripheral blood cell proliferation. Abnormal cell metabolism is a new feature of malignant proliferation of tumor cells, but the role of metabolism in the pathogenesis and prognosis of PV remains unclear. We analyzed metabolic differences of peripheral blood sera between 32 PV patients and 20 healthy controls (HCs) by liquid chromatography–mass spectrometry (LC–MS) to investigate their relationship with cell proliferation and to screen for prognosis-related metabolic biomarkers. Compared to HC, 33 endogenous metabolites were significantly changed in PV and were involved in fatty acid metabolism, glucose metabolism, sphingolipid metabolism, and amino acid metabolism pathways. Among them, seven metabolites were closely associated with JAK2 mutations, 2 of which may contribute to the proliferation of peripheral blood cells in PV patients. A set of potential prognostic metabolic biomarkers containing four metabolites was identified by a receiver operating characteristic (ROC) curve according to the risk stratification of the PV patients and their combined AUC value of 0.952, with a sensitivity of 90.905% and specificity of 90.909% at the optimal cutoff point. Metabonomics is an important tool for the study of the pathogenesis of PV and the relationship between JAK2 gene mutation. Furthermore, the potential biomarkers of this study may provide a reference for the prognosis of PV.

目的 探索益气温阳汤治疗慢性免疫性血小板减少症(chronic immune thrombocytopenia,CITP)的免疫调控机制及转录因子GATA结合蛋白(GATA binding protein,GATA)-3通路对血小板计数的预测价值.方法 纳入28例CITP患者为治疗组,20名健康志愿者为健康对照组,治疗组给予益气温阳汤治疗,健康对照组不给予干预,4个月后用流式细胞术检测治疗前后外周血T、B淋巴细胞亚群及辅助性T细胞(helper T cell,Th)1、Th2水平,荧光定量PCR检测T-bet、GATA-3、Foxp3 mRNA表达水平,Logistic回归分析法对血小板相关预测因素进行分析.结果 益气温阳汤治疗CITP总有效率42.86％,与治疗前比较,治疗组治疗后CITP患者CD3+总T细胞、细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)、自然杀伤T细胞(natural killer T cell,NKT)、GATA-3 mRNA表达水平显著降低,血小板、CD5+B淋巴细胞比值、调节性T细胞(regulatory T cell,Treg)、Th2细胞水平升高(P<0.05);治疗组治疗后Foxp3 mRNA表达水平显著低于健康对照组(P<0.05).Logistic回归分析显示,治疗后GATA-3水平与血小板呈负相关,方程式:血小板=90.299-64.086×(GATA-3).结论 益气温阳汤通过调节CITP患者T、B淋巴细胞亚群水平,起到免疫调节和免疫耐受作用,并可以通过调控GATA-3和Foxp3 mRNA表达提升血小板计数,GATA-3 mRNA的表达水平可能对CITP患者血小板计数有预测价值.

ObjectivesThe currently recommended aspirin regimen appears inadequate for thromboprophylaxis in essential thrombocythemia (ET). This study aimed not only to evaluate the curative effect of aspirin but also to explore the coagulation status and determinants of aspirin resistance (AR) of ET patients.MethodsA total of 80 ET patients who underwent coagulation tests, thromboelastography (TEG), and next-generation sequencing (NGS) were involved in the study. Patients were divided into the aspirin sensitivity (AS) group and AR group according to the arachidonic acid inhibition rate. Their clinical features and coagulation function were analyzed.ResultsThe incidence of AR was 53.75% (43/80) in 80 ET patients. Fbg was significantly higher in coagulation tests in AR patients compared with AS patients (P < 0.05), while the differences in other variables (D-D, PT, PTA, INR, APTT, TT, FDP, and AT-III) were not statistically significant (P > 0.05). Compared with AS patients, the K values, α angles, MA values, and CI values of TEG in AR patients were statistically smaller (P < 0.05), but there was no significant difference in R value between them (P > 0.05). Univariate and multivariate logistic regression analysis showed that age, irregular use of aspirin, smoking, dyslipidemia, and hypertension increased the risk of AR (P < 0.05). In the routine NGS, the driver gene and non-driver gene had no effect on AR in ET patients.ConclusionCompared with AS patients, AR patients have enhanced platelet aggregation function, are in a relatively hypercoagulable state, and haveelevated fibrinogen function/levels, all of which cause a worse coagulation status. ET patients with increasing age, irregular use of aspirin, smoking, dyslipidemia, and hypertension are possibly at higher risk of AR. The routine NGS may not be helpful for the prediction of AR, therefore we recommend adding relevant drug-resistance genes to NGS.

Background: Polycythemia vera (PV) is a refractory hematological disease that lack of effective therapy. Chinese traditional medicine Longchai Jiangxue formula (LCJX) has showed the powerful effects on PV. However, the active ingredients and mechanisms of this formula have not been elucidated. We explored the active ingredients and mechanisms of LCJX for treating PV. Methods: The chemical constituents of LCJX were qualitatively analyzed by UPLC/Q-TOF-MS/MS. On this basis, the TCMSP, ETCM, PubChem BioAssay and ChEMBL databases were searched to predict the potential targets of chemical components of LCJX. Then Genecards, GEO, DisGeNET, and OMIM databases were used to retrieve data of targets related to PV. Drug-disease-target network and protein-protein-interaction (PPI) network were built. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed. Finally, Molecular docking, CCK-8 assay, Annexin V-FITC/PI staining and western blot were processed so as to screen the active components related to PV and elucidate its mechanisms. Results: A total of 84 compounds were identified from LCJX by UPLC/Q-TOF-MS/MS. After removed duplicate items, there were 143 targets linked to both disease and drugs. Crucial genes, such as MTOR, HIF1A, JAK2, VEGFA, STAT3, AKT1, TERT, MAPK1, were shown in PPI network. GO enrichment indicated that oxidative stress process, tyrosine kinase activity and phosphatase binding function, and cell membrane structure were in reference to LCJX against PV. KEGG enrichment showed that JAK-STAT signaling pathway and PI3K-Akt signaling pathway, were put in an important position of the treatment. Furthermore, Molecular docking, CCK8 assay, Annexin V-FITC/PI staining and western blot technique proved the therapeutic effect of Saikosaponin A, main ingredient of LCJX. Conclusion: This study, combined with UPLC/Q-TOF-MS/MS, network pharmacology and molecular biology, provides a reference for the identification of effective components, screening of quality markers and analysis of its action mechanism of LCJX.

肺癌发病机制是在各种病因影响下,肺脏生理功能失调,气血津液运行失常,产生以痰邪为主的病理产物.痰邪蕴结于肺,日久渐积难消而恶变,发展为肺癌.痰邪的流窜性可能与肺癌细胞的转移有重要关系.治疗肺癌应以理气化痰为基本原则.二陈汤理气健脾、燥湿化痰,药少而力专,是治疗痰证的经典方剂,可通过调节细胞免疫来防治肺部恶性肿瘤,亦可提高肺癌铂类化疗方案的疗效,对肺癌转移具有抑制作用,减轻肺癌患者临床症状,提高患者化疗后生活质量.

OBJECTIVE:To investigate the relationship between JAK2 gene mutation and clinical indicators in patients with myeloproliferative neoplasms (MPN).METHODS:122 MPN patients in the Department of Hematology, Xiyuan Hospital, China Academy of Chinese Medical Sciences from September 2017 to January 2020 were retrospectively analyzed. The relationship between JAK2 gene mutation and sex, age, peripheral blood cell count, splenomegaly, and thrombosis and bleeding events were analyzed.RESULTS:In 122 patients with MPN, the patients with polycythemia vera (PV) accounted for 36 (29.5%), the patients with essential thrombocythemia (ET) accounted for 56 (45.9%), the patients with myelofibrosis (MF) accounted for 30 (24.6%). The JAK2 gene mutation rate in MPN patients was 64.6% (79/122), and the JAK2 gene mutation rate in PV, ET and MF groups were 77.7% (28/36), 60.7% (34/56) and 56.7% (17/30), the JAK2 gene mutation rate of the patients in PV group was statistically significant as compared with those in the ET group (P＜0.05). The hemoglobin (Hb) count of the patients in JAK2 gene mutation group was higher than those in wild-type group ［(150.0±39.6)g/L vs (129.4±38.9)g/L, P＜0.05］; the white blood cell (WBC) count of the patients in JAK2 gene mutation group was higher than those in the wild type group ［(9.5±4.7)×109/L vs (8.4±46.9)×109/L, P＜0.05］. As for the patients in PV group, the platelet count of the patients in JAK2 gene mutation group was higher than those in the wild type group ［(370.2±113.1)×109/L vs (264.8±63.9)×109/L, P＜0.05］. The incidence of splenomegaly in MPN patients was 35.2% (43/122), and the incidence of splenomegaly in MF patients was 63.3% (19/30), and the incidence of splenomegaly in the patients in JAK2 gene mutation group in MF group (82.4%, 12/17) was significantly higher than those in the wild-type group (38.5%, 5/13) (P<0.05).CONCLUSION:The mutation rate of JAK2 gene in MPN patients is higher, and the mutation rate of JAK2 gene in PV patients is higher than that in ET and MF patients; JAK2 gene mutations in MPN patients are related to hemogram index; the incidence of splenomegaly is the highest in MF patients, and splenomegaly is related to the occurrence of JAK2 gene mutations in MF patients.

目的:基于数据挖掘分析王沛教授治疗胰腺癌的用药规律.方法:收集北京中医药大学东方医院肿瘤科门诊,2010年9月10日至2018年1月12日在王沛教授处就诊的胰腺癌患者资料.采用频数统计,网络图,Apriori算法等对处方进行分析.结果:纳入处方283首方,共涉及药物164味,最常用的药物为生半夏、瓜蒌和黄芪等,关联分析显示相关性较好的药物为薤白、瓜蒌、生半夏和黄连等.结论:王沛教授治疗胰腺癌多用补虚、化痰、理气之品.

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 < 0.00001) and a higher probability of transformation to acute leukemia (LFS: HR = 1.77, 95% CI: 1.30–2.42, P = 0.0003; the rate of acute leukemia transformation: OR = 2.06, 95% CI: 1.50–2.83, P < 0.00001). Furthermore, ASXL1 mutations were correlated with patients older than 65 years old, male, a lower level of platelet counts, and a higher risk of the international prognostic score system. These findings indicate that ASXL1 mutations have a significant adverse impact on the prognosis of PMF patients and may contribute to risk stratification and prognostic assessment for PMF patients.

目的 系统评价滋阴清热法预防性干预头颈部恶性肿瘤急性放射性口腔黏膜炎的临床疗效.方法 通过计算机检索中国知网(CNKI)、万方数据知识服务平台、中国科技期刊数据库(VIP)、中国生物医学文献数据库(CBM)、PubMed、Cochrane Library数据库,检索时限为自建库至2019年12月30日.2位研究者严格按照纳入标准独立筛选提取文献,根据Cochrane标准进行文献纳入筛选、数据提取、质量评价,采用RevMan 5.3软件进行数据分析.结果 最终共纳入18项随机对照试验(RCT),合计1341例患者,Meta分析结果显示,观察组急性放射性口腔黏膜炎发生时间较对照组明显延后[MD = 6.97,95％CI(3.71,10.22),P＜0.0001],口腔黏膜Ⅲ～Ⅳ级损伤发生率明显低于对照组[OR=0.17,95％CI(0.13,0.22),P＜0.00001],治疗有效率明显高于对照组[OR = 8.25,95％CI(4.83,14.08),P＜0.00001].结论 滋阴清热法预防性干预,能延迟放射性口腔黏膜损伤出现时间,明显降低Ⅲ～Ⅳ级口腔黏膜损伤发生率,治疗有效率高于西医常规治疗,由于本研究纳入的RCTs质量和数量原因,临床需更多高质量的RCTs研究对滋阴清热法防治急性放射性口腔黏膜炎的临床疗效进行验证.

[目的]基于数据挖掘和网络药理学方法,探讨胡晓梅主任治疗真性红细胞增多症的用药规律及其作用机制,为真性红细胞增多症的治疗提供新思路.[方法]通过回顾性研究方法,收集2018年4月1日—2020年11月30日胡晓梅主任治疗真性红细胞增多症(PV)的有效方剂202首,通过中医传承辅助平台(V2.5)软件分析其处方的用药规律,通过频次分析筛选累计频次最高的前10味中药作为其治疗本病的核心药物.然后对核心药物进行了网络药理学分析,探索其治疗PV的潜在的分子机制.[结果]通过药物频次分析得到的前10位的核心药物是生地黄、柴胡、白花蛇舌草、赤芍、龙葵、半枝莲、龙胆、夏枯草、黄芩、牛膝.用药模式分析结果显示最常见的核心组合是生地黄和柴胡.网络药理学分析结果显示核心药物的靶点为378个,与PV相关的疾病靶点为589个,韦恩图显示核心药物与PV相关的靶点为114个.基因本体论(GO)富集分析显示核心药物治疗PV主要涉及氧化应激、药物反应及细胞凋亡等生物过程,它们通过细胞因子受体-配体结合、磷酸化作用、血红素结合、蛋白结合、激酶调节等分子功能在细胞膜、分泌囊泡等部位发挥作用.京都基因与基因组百科全书(KEGG)富集于PI3K-Akt、HIF-1、JAK-STAT、P53等信号通路.蛋白互作网络图显示,STAT3、AKT1、TP53、MAPK1、TNF、VEGFA、MAPK14、IL6有可能是核心药物治疗PV的关键靶点.[结论]运用数据挖掘和网络药理学分析方法,获得胡晓梅主任治疗PV的高频核心药物,并揭示了核心药物具有多成分、多靶点、多通路治疗PV的潜在作用机制,为临床治疗PV提供了参考方案,并为进一步深入的物质基础和分子机制研究提供了思路.