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Intracranial hemorrhage (ICH) is a devastating complication of immune thrombocytopenia (ITP). However, information on ICH in ITP patients under the age of 60 years is limited, and no predictive tools are available in clinical practice. A total of 93 adult patients with ITP who developed ICH before 60 years of age were retrospectively identified from 2005 to 2019 by 27 centers in China. For each case, 2 controls matched by the time of ITP diagnosis and the duration of ITP were provided by the same center. Multivariate analysis identified head trauma (OR = 3.216, 95%CI 1.296–7.979, P =.012), a platelet count ≤ 15,000/μL at the time of ITP diagnosis (OR = 1.679, 95%CI 1.044–2.698, P =.032) and severe/life-threatening bleeding (severe bleeding vs. mild bleeding, OR = 1.910, 95%CI 1.088–3.353, P =.024; life-threatening bleeding vs. mild bleeding, OR = 2.620, 95%CI 1.360–5.051, P =.004) as independent risk factors for ICH. Intraparenchymal hemorrhage (OR = 5.191, 95%CI 1.717–15.692, P =.004) and a history of severe bleeding (OR = 4.322, 95%CI 1.532–12.198, P =.006) were associated with the 30-day outcome of ICH. These findings may facilitate ICH risk stratification and outcome prediction in patients with ITP. [ABSTRACT FROM AUTHOR]

RP215 monoclonal antibody (Mab) was initially generated against OC-3-VGH ovarian cancer cells and was shown to react with a cancer-associated carbohydrate epitope in glycoproteins designated as CA215. Additional five high affinity Mabs, designated as RCA-10, -100, -104, -110 and -111, respectively, were generated by using affinity-purified CA215 as the immunogen in this study. All RCA Mabs were found to recognize periodate-sensitive carbohydrate-associated epitope(s) and to pair with RP215 in typical sandwich enzyme immunoassays for the quantification of CA215. When compared with those of RP215, the amino acid sequence homology of the Fab regions ranged from 100%% for RCA-100 to 65%% for RCA-110, based on which 3 distinct Mab groups were categorized. In vitro TUNEL apoptosis and complement-dependent cytotoxicity assays were performed with these Mabs and found to have comparable inhibitory efficacy to cancer cells. Results of biochemical and immunological assays revealed that RP215, RCA-100 and RCA-10 react with the linear carbohydrate-associated epitope, whereas the others recognize the conformational form of the epitope in CA215. This study has suggested that the unique carbohydrate-associated epitope(s) is immunodominant in mice when immunized with CA215. It remains to be demonstrated if the differential anti-cancer efficacy exists among the distinct groups of these anti-CA215 Mabs. [ABSTRACT FROM AUTHOR]

Objective: To study the expression level of TGFβ1 and VEGF gene in patients with acute myeloid leukemia (AML) and its clinical prognostic value., Methods: Seventy-eight AML patients treated in our hospital from July 2016 to September 2018 were selected. After isolation of bone marrow mononuclear cells from the patients, the levels of TGFβ1 and VEGF genes were detected by RT-PCR, and the correlation of TGFβ1 with VEGF genes and clinical characteristics of AML patients was analyzed. OS and EFS of the patients were evaluated by Kaplan-Meier, and Cox risk ratio model was used to analyze the prognostic risk factors of AML patients., Results: The relative expression level of TGFβ1 gene in AML patients was 0.32±0.04, which was significantly lower than that in control group (P<005). The relative expression level of vascular endothelial growth factor(VEGF) gene in the patients was 2.65±0.15, which was significantly higher than that in the control group (P<0.05). The levels of TGFβ1 and VEGF genes significantly correlated with leukocyte count, hemoglobin, platelet and peripheral blast levels in AML patients (P<0.05). The level of TGFβ1 in AML patients with complete remission was higher than that in patients with partial remission or non-remission (P<0.05). The level of TGFβ1 in AML patients with partial remission was significantly higher than that in patients with non-remission (P<0.05). The level of VEGF in AML patients with complete remission was lower than at in patients with partial remission or non-remission (P<0.05). The level of VEGF in AML patients with partial remission was significantly lower than that in patients with non-remission (P<0.05). Kaplan-Meier survival analysis showed that OS and DFS in AML patients with high expression of TGFβ1 were better than those in patients with low expression of TGFβ1 (P<0.05), OS and DFS in AML patients with low expression of VEGF were better than those in patients with high expression of VEGF (P<0.05). Multivariate Cox regression analysis showed that platelet, TGFβ1 and VEGF gene were independent influencing factors of OS (P<0.05). Leukocyte, TGFβ1 and VEGF gene were independent influencing factors of DFS (P<0.05)., Conclusion: Decreased expression of TGFβ1 and increased expression of VEGF gene in AML patients closely relate to the poor prognosis of AML patients, which can provide reference for improving clinical efficacy of AML patients.

Objective: To investigate the expression of CD44, CD87 and CD123 in acute leukemia and its correlation with cellular immune markers., Methods: A total of 166 patients with acute leukemia (AL) admitted from May 2014 to February 2017 were enrolled in AL groups. Among these patients, 100 patients suffered from acute myeloid leukemia, 50 patients suffered from acute lymphoid leukemia, and 16 patients showed B/medullary phenotype. At the same time 50 patients with non-acute leukemia were enrolled in the control group. 5 ml of fasting venous blood collected from the patients in each group, and the percentage of CD44, CD87 and CD123 cells was determined by three-color flow cytometry. Symptomatic chemotherapy was given to the patients with confirmed acute leukemia, and the remission was evaluated after 2 treatmen courses. The Complete remission (CR) was recorded and the percentage of CD44, CD87 and CD123 cells under different curative efficacy were recorded. The correlation of the prognosis patients with CD44, CD87 and CD123 was analyzed by SPSS Pearson correlation analysis software., Results: The positive rates of CD44, CD87 and CD123 in AL group were all higher than those in the control group (P＜0. 05). The positive rates of CD44 and CD123 in acute myeloid leukemia group were higher than those in acute lymphoblastic leukemia group and B/myeloid phenotype group (P＜0. 05). The positive rate of CD44 in acute lymphoid leukemia group was higher than that in B/medullary double phenotype group (P＜0.05). The treatment in the patients of AL group was successfully completed. 132 patients reachel to CR and 34 patients to PR+NR after 2 courses. The positive rates of CD44, CD87 and CD123 in CR patients were lower than those in PR+NR patients (P＜0.05). The results of SPSS Pearson correlation analysis showed that the prognosis of patients with acute leukemia negatively correlated with CD44 and CD87 (P＜0.05)., Conclusion: The expression of CD44, CD87 and CD123 in different phenotype of acute leukemia are different, which correlateds with prognosis. The determination of CD44, CD87 and CD123 can be used to evaluate the prognosis of patients for the reference of clinical treatment.

Objective: To analyze the prognostic value of BCL-2, BCL-6 and MYC in patients with diffuse large B cell lymphoma (DLBCL)., Methods: One hundred and sixty three cases of DLBCL in our hospital from March 2012 to March 2015 were selected. The specimens of lymphoma tissue of patients were collected. The expression of BCL-2, BCL-6 and MYC was detected by immunohistochemical method. The fusion of IGH/BCL-2, the gene breakage of BCL-6 and MYC were detected by interphase fluorescence in situ hybridization. The correlation of the expression levels of BCL-2, BCL-6 and MYC with the clinicopathological features and prognosis in the patients with DLBCL was further analyzed., Results: MYC, BCL-2 and BCL-6 showed pale brown or reddish brown positive signals, among them MYC mainly positively expressed on the cell membrane, and BCL-2 mainly expressed on the cytoplasm and local cell membrane, and BCL-6 mainly expressed in the nucleus. The expression level of BCL-2 in ECOG physical status score 2 was higher than that in patients with ＜2 scores, and the expression level of BCL-2 in CD5 + and germinal center B-cell-like (GCB) was significantly higher than that in patients with non-GCB (P＜0.05), and the international prognostic index (IPI) for 3-5 scores at the MYC expression level was significantly higher than that of the 0-2 score (P＜0.05); the expression level of BCL-6 in immune subtype CD5 + and GCB was significantly lower than that in non-GCB (P＜0.05). The results of Cox multivariate analysis showed that the expression level of BCL-2, BCL-6 and MYC significant correlate with the overall survival and progression-free survival (P＜0.05) of the patients with DLBCL., Conclusion: BCL-2, BCL-6 and MYC as important molecular markers are of high value for evaluating the prognosis of patients with DLBCL.

Objective: To investigate the clinical features of β-thalassaemia intermediate (TI) patients and the curative effect and side reactions of hydroxyurea therapys., Methods: Twenty nine patients with TI were divided into hydroxyurea therapy group and no hydroxyurea therapy group; the curative effect and side reactions in 2 groups were compared; the situation of blood transfusion in the 2 groups was evaluated., Results: In hydroxyurea therapy group, the hemoglobin level increased after treatment for 3 months; the reticulocyte percentage obviously decreased after treatment for 12 months; the serum ferritin had been maintained at a low level; while in no hydroxyurea therapy group, the levels of hemoglobin and reticulocytes were not significantly improved after treatment, the serum ferritin level gradually increased. In hydroxyurea therapy group, 12 cases were out of blood transfusion after treatment for 12 months, effective rate of treatment was 85.71%; while in no hydroxyurea therapy group, the blood transfusion dependency was not improved after treatment. No serious side reactions were found in all the hydroxyurea treated patients., Conclusion: The hydroxyurea shows a better curative effect on TI patients, no serious side reactions occur in all the patients treated with hydroxyurea, but the long-term curative effect and side reactions should be observed continuously.

Objective: To investigate the relationship between the etiology, laboratory findings and prognosis of secondary HPS, so as to enhance the understanding of the secondary HPS and the related factors affecting prognosis, reduce the misdiagnosis and to understand the factors that affect the prognosis., Methods: The etiology, laboratory findings and prognosis of 61 patients with secondary HPS were analyzed retrospectively., Results: Univariate analysis showed that TG, FIB, SF, ANC, ALB, TBIL, ALT, LDH were significantly different between the 2 groups of the patients with secondary HPS. Multiariate factor analysis showed that the LDH and the etiology affected the prognosis of the patients with secondary HPS. The prognosis of the patients with elevated LDH, viral infection, especially EB virus infection, tumor and unknown causes might be poor., Conclusions: The etiology and clinical characteristics of prognosis are diverse. The cause needs to be identified as soon as possible. The prognosis should be judged according to LDH and other indicators. Then, targeted therapy should be used to control the disease in the short time.

Some cases of myeloproliferative disorders (MPD) could be transformed into acute myeloid leukemia (AML), but the cell differentiation process of MPD into AML has not yet been observed in vivo. This study was aimed to reveal this differentiation process. The flow cytometry was used to analyse the immunophenotype of differentiated cells of 2 MPD cases who developed into AML in a short time. The reports showed that the different MPD-AML subclones are presented when the MPD cells that proliferate slowly in vivo become the AML blast cells that proliferate rapidly. It is concluded that understanding the process of MPD crisis will help the MPD-AML early diagnosis and treatment.

Objective: To study the risk factors of bronchial asthma of Li nationality in Hainan., Methods: A total of 13 050 subjects of Li nationality were selected by random unequal ratio stratified cluster sampling method from southern, central and western part of Hainan and investigated with Hainan Epidemiological Asthma Survey Questionnaire of Li Nationality. There were 441 cases of bronchial asthma, and 1296 cases of control that were sampled by random number table method. The logistic regression method was used to analyze risk factors., Results: The asthma prevalence of Li nationality in Hainan was 3.38%(441/13 050). The main risk factors of asthma were family asthma (OR = 4.323, 95%CI = 3.259 - 5.735), hypersensitiveness (OR = 7.775, 95%CI = 5.686 - 10.632), smoking (OR = 1.494, 95%CI = 1.174 - 1.902), cooking fuels and living environment. Cold air change (OR = 1.604, 95%CI = 1.286 - 2.001) and respirable dust or irritant gas (OR = 2.123, 95%CI = 1.702 - 2.648) were the important incentives., Conclusion: The main risk factors of asthma among Li nationality were family asthma, hypersensitiveness, smoking, cooking fuels by means of fuel oil, hay or wood, living environment by means of couch grass room and human-livestock mix live, cold air change, respirable dust or irritant gas.

Objective: To study the effect of Thromboxane A2 (TXA2) inhibitor on the proliferation of hypoxic pulmonary artery smooth muscle cells (PASMC) of porcines., Methods: The methods of 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), immunohistochemistry of Ki67 and TdT-mediated dUTP nick end labeling (TUNEL) were employed to measure the proliferation, inhibition rate and apoptosis of PASMC., Results: U46619, a thromboxane A2 agonist, could promote the proliferation and expression of Ki67 in PASMC under hypoxic and normal conditions. It could also inhibit the apoptosis of PASMC. Ozagrel, a thromboxane A2 inhibitor, inhibited the proliferation and the expression of Ki67 in PASMC under hypoxia. The inhibition rate was 71.4%, but it had no effect on the proliferation and expression of Ki67 in PASMC under normal conditions. It promoted the apoptosis of PASMC. Ozagrel could inhibit the action of U46619 in promoting the proliferation and expression of Ki67 in PASMC and inhibit the apoptosis of PASMC., Conclusion: Ozagrel may be used in treating the pulmonary hypertension by inhibiting the proliferation of PASMC, pulmonary vascular remodeling, and promote the apoptosis of PASMC.

Objective: To learn the prevalence rate and distribution of bronchial asthma of Li nationality in Hainan by epidemiological survey so as to provide rationales for preventing and treating asthma., Methods: A total of 13 050 subjects of Li nationality in Hainan were investigated by random sampling of stratified cluster method with an unequal ratio according to the medical records, physical signs and symptoms and pulmonary function tests. Then the prevalence was analyzed according to different population characteristics., Results: The prevalence rate of asthma was 3.38% for Li nationality in Hainan. And it increased with an advancing age. It was much higher in agricultural population (4.08%) than that in cultivation and commercial populations (2.52% and 1.97%, P < 0.01). The asthma prevalence rate was in the highest at 4.62% in rubber workers. And it was much higher in rural areas than that in urban areas (3.85% vs 2.63%, P < 0.01)., Conclusions: The epidemiological distribution of asthma of Li nationality in Hainan has been determined. And asthma remains a severe problem locally. The efforts of preventing and treating asthma and its education should be strengthened.

Objective: To investigate the synergistic therapy effects of B and T lymphocyte attenuator (BTLA) extracellular domain in combination with heat shock protein 70 (HSP70)-TC-1 antigen peptide complex on the mouse model of cervical cancer and the related immunological mechanisms., Methods: (1) Detecting the BTLA and herpesvirus entry mediator (HVEM) gene expression in the tumor microenvironment after C57BL/6 mice were inoculated with TC-1 tumor cells by realtime PCR; BTLA, HVEM expression on tumor infiltrating lymphocytes cell surface were detected by flow cytometry (fluorescence intensity). (2) According to different treatments, tumor-bearing mice were divided into 5 groups, which was injected with pcDNA3.1 (empty vector plasmid as control), psBTLA (vector plasmid which expresses BTLA extracellular domain), HSP70 (HSP70-TC-1 cell peptide complex), HSP70 + pcDNA3.1 or HSP70 + psBTLA, respectively. The weight of tumor was recorded. The expression of immunoregulatory genes in tumor microenvironment were detected. The change of lymphocyte amount and cytotoxicity were detected too; lymphocyte proliferation activity was measured by tritium thymidine incorporation assay; the concentration of interleukin (IL)2 and interferon-γ (IFN-γ) in supernatants of spleen lymphocyte were measured by enzyme-linked immunosorbent assay (ELISA)., Results: (1) BTLA gene expression was gradually increased after tumor cells inoculation. The highest expression level was 2.83 ± 0.35 at 14th day, which had statistical significance difference with the 7th day expression of 1.66 ± 0.25 (P < 0.05). While HVEM mRNA expression did not change significantly (P > 0.05). The 7th and 14th day after TC-1 cells inoculation, the average fluorescence intensity of BTLA expression on the surface of tumor infiltrating lymphocytes was 33.5 and 51.8, respectively, in which there was statistically significant difference (P < 0.05); while the difference of HVEM expression was not statistically significant (57.2 vs 49.3, P > 0.05). (2) The 28th day after inoculation, tumor inhibition rate of HSP70 + psBTLA group was 88%, which was significantly higher than other treatment groups (P < 0.05). The 28th day after TC-1 cells inoculation, combination therapy not only promoted IFN-γ and IL-2 gene (3.12 ± 0.71, 3.20 ± 0.62) expression but also reduced transforming growth factor-β (TGF-β), Foxp3 and IL-10 expression (0.25 ± 0.03, 0.19 ± 0.03, 0.31 ± 0.04; P < 0.05). It also promoted CD₈(+) T lymphocyte infiltration (52 ± 6)/high power field, cytotoxicity (65.5 ± 2.4)%, proliferation (15.0 × 10³ cpm) and cytokine IL-2, IFN-γ secretion (824 ± 51), (1096 ± 112) pg/ml, which were all significantly higher than other groups (P < 0.05)., Conclusion: The effect of immunotherapy on tumor can be augmented by the combination of psBTLA which expresses extracellular domain of BTLA and HSP70-TC-1 tumor antigen peptide complex, which could improve the expression of the related immunoregulatory genes to establish a much better microenvironment in favor of anti-tumor immune response against the mice model of the cervix carcinoma.

Objective: To observe the influence of the plasma thromboxane B2 (TXB2), 6-keto-PGF1alpha, CD62P and PAC-1 and Thrombus in patients with primary thrombocytosis (ET). To observe the effect of sodium ozagrel to prevent and treat thrombosis in patients with ET., Methods: The subjects including 48 patients with ET. All patients were measured the plasma TXB2, 6-keto-PGF1alpha, CD62P and PAC-1 before and after treatment with or without sodium ozagrel., Results: The plasma levels of CD62P, PAC-1, TXB2, 6-keto-PGF1alpha and TXA2/PGI2 in the patients with ET were significantly higher than the normal people (P < 0.01). The levels of CD62P, PAC-1, TXB2, TXB2/6-keto-PGF1alpha in patients with treatment of sodium ozagrel were higher than patients without treatment of sodium ozagrel (P < 0.01). The plasma levels of CD62P, PAC-1 and TXA2/PGI2 in patients with treatment of sodium ozagrel and that in normal people had no significant distinction (P < 0.01). All the index of conventional therapy group were higher than normal people (P < 0.01) but had no significant distinction with the patients before conventional treating. The incidence of thrombus in patients treated with sodium ozagrel was lower than patients treated without sodium ozagrel (P < 0.05)., Conclusion: With the treatment of sodium ozagrel in patients with ET, the CD62P, PAC-1, TXB2 and TXA2/PGI2 of plasma could be decreased. And the incidence of thrombus was decreased.

This study was aimed to investigate the incidence of thrombosis in patients with primary thrombocytosis (PT) and its correlation with function changes of platelets, and to explore the effect of thromboxane A2 (TXA2) inhibitor-ozagrel sodium on platelet activity and its efficacy for prevention and treatment of thrombosis. The CD62P and PAC-1 levels on platelet surface were detected by flow cytometry; the levels of TXB2 (metabolic product of TXA2) and 6-keto-PGFIalpha (metabolic product of prostacyclin) were detected by FLISA. The function change of platelets and its correlation with thrombosis were observed and compared in PT patients with and without thrombosis. The results indicated that the TXB2, PAC-1 and CD62P level, and TXB2/6-keto-PGF1alpha ratio in PT patients with thrombosis were higher than those in PT patients without thrombosis before treatment with ozagrel sodium (p<0.01). After treatment with ozagrel sodium, the function indexes of platelets such as CD62P, PAC-1, TXB2 and TXB2/6-keto-PGF1alpha except 6-keto-PGF1alpha in PT patients with and without thrombosis decreased obviously (p<0.01), but there was no significant difference in TXB2, 6-keto-PGF1alpha and TXB2/6-keto-PGF1alpha levels between PT patients with and without thrombosis except CD62P and PAC-1. It is concluded that the multi-index of platelets in PT patients with thrombosis are higher than that in PT patients without thrombosis, the activation of platelet function is a high risk factor for thrombosis of PT patients. The ozagrel sodium can obviously reduce the platelet activation, decrease the production of TXA2 and ameliorate the TXB2/6-keto-PGF1alpha ratio. The ozagrel sodium not only possesses therapeutic effect, but also preventive efficacy for thrombosis.

Objective: To evaluate of liquid based cytology test (LCT) in avoiding medical tangles., Methods: One thousand five hundred five thirty one cases, which were collected from out-patients of precancerous lesions of uterine cervix, were randomly divided into three groups based on different smear preparation: LCT method was used in two groups (one with ThinPrep kit and one with ArtoBrain kit), conventional Papauicolaou smear (PS) was used in one group. All cases of abnormal cervical smears were identified by cytologic test underwent colposcopic examination and colopscopically multiple biopsy. Results of test were analyzed by software SPSS 11.0., Results: Significant diference were found between LCT method and PS method compared by index of satifacation, sensitivity, specificity, accuracy, false negative rate and erroneous diagnosis rate (P < 0.05, but no difference were found between two LCT groups (ThinPrep kit and ArtoBrain kit)., Conclusion: LCT method can improve diagnostic level of precancerous lesions of uterine cervix either tested by ThinPrep kit or ArtoBrain kit, so have the powerfull value to avoid medical tangles.

Objective: To evaluate the effect of intracytoplasmic sperm injection (ICSI) using cryopreserved-thawed testicular spermatozoa with testicular fine needle aspiration (TEFNA) in patients with non-obstructive azoospermia., Methods: Sixty-two patients underwent TEFNA. Mature testicular spermatozoa were found in 35 cases of patients and the testicular tissues were cryopreserved for later ICSI. Ovarian stimulation included a long protocol of GnRHa/FSH/hCG. Oocyte retrieval was performed under transvaginal ultrasound guidance, and ICSI conducted with cryopreserved-thawed testicular spermatozoa., Results: A total of 35 couples underwent 35 ICSI cycles using cryopreserved-thawed testicular spermatozoa with testicular fine needle aspiration. The clinical pregnancy rate was 37.14% (13/35)., Conclusion: ICSI using cryopreserved-thawed testicular spermatozoa with testicular fine needle aspiration is a main and effective method in the treatment of non-obstructive azoospermia, which can avoid further testicular fine needle aspiration.

immunosorbent To determine serum EPO and soluble transferrin receptor (sTfR) level and to assess the degree of erythropoietic proliferation in patients with thalassemia of Li nationality, 50 cases and 50 normal individuals were studied. Enzyme-linked immunosorbent assay was used for EPO and sTfR. Results showed that serum EPO and sTfR levels in beta-thalasemia major patients were significantly higher than those in normal control group, and no significant difference in beta-thalassemia minor cases with those in control. It is concluded that increase of serum EPO and sTfR level is related to the type of thalassemia.

Objective: To explore a stable technology for inducing ES cell to committed hematopoietic differentiation., Methods: The effects of various inductive factors on BLast Colony-Forming Cell (BL-CFC) were investigated. In vitro differentiative system for ES cells was employed in this study., Results: A high linear correlation between the number of D3.5 EB-derived cells plated and the number of blast cell colonies was developed, r = 0.9931. With high frequency of blast colonies observed (1.08-1.2 colonies per 100 cells). 20%-30% D4T conditioned medium (D4T CM) showed the most significant growth potentials of blast colonies. D4T CM, EPO or KL alone had no blast colony growth promoting effect (P > 0.05). But VEGF alone had high significant blast colony growth promoting effect (P < 0.001). However, any two factors combination from above four factors exerted better growth promoting effect than VEGF alone (EPO + D4T CM, P < 0.05; KL + D4T CM, P < 0.01; VEGF + D4T CM, P < 0.001). There were no significant difference among VEGF + KL and EPO + D4T CM or KL + D4T CM, and KL + D4T CM (P > 0.05). While the combination of VEGF + D4T CM was better than KL + D4T CM, VEGF + KL or EPO + D4T CM (P < 0.001). Moreover, the combination of VEGF + KL + D4T CM + EPO, had the highest significant blast colony growth promoting effect (P < 0.001). And the highest frequency of blast colonies was observed (1.5-1.2 colonies per 100 cells)., Conclusion: VEGF may be the main factor which stimulates the growth of significant numbers of blast cell colonies. D4T CM maybe contains strong cofactors. EPO and KL are the main factors for the induction of BL-CFC to committed hematopoietic differentiation. D3.5 EB-derived cells are more sensitive to various stimulators and have strong blast colony growth promoting effect than that of D3.25 EB-derived cells.