ObjectiveTo compare the diagnostic value of magnetic resonance imaging (MRI),computed tomography (CT) and radiography in the early detection of arthropathies of haemophilia.Methods Prospective studies of 21 joints in 11 patients were studied with X-ray,CT and MR examination. The 21 joints with haemophilia arthropathies were divided into three groups according to Pettersson scoring system.0 point were the first group,＜4 points were the second group,≥4 points were the third group.Abnormal imaging findings of osteoporosis,enlarge epiphysis,erosion of cartilage,irregular subchondral surface,narrowing of joints space,joint deformity,subchondral cyst formation,effusion/haemarthrosis of joint,synovial hypertrophy with haemosiderin,deformity of joints were used for all imaging comparison.The results were analysis with Chi-square test.To compare the first group,irregular subchondral surface and the number of subchondral cyst formation of all symptomatic joints were detected by CT and MR,the results were analysis with pair-sample t test.ResultsModerate and severe hemophilic joints were found in 80.95％ (17/21)of twenty-one symptomatic joints,and mild hemophilic joints were found in 19.05％ (4/21).The detected results were the same in enlarge epiphysis,narrowing of joints space,joint deformity in all joints by radiography,CT and MR.Significant difference in detection of irregular subchondral surface,subchondral cyst formation,effusion/haemarthrosis of joint,were found between radiography with either CT (x2 value 19.06,16.70,4.84,P ＜0.05 ) or MRI (x2 value 19.06,16.70,7.76,P ＜0.05),Significant difference in detection of the first group joint irregular subchondral surface and the subchondral cyst formation total number were found between CT and MR ( x2 =3.29,P ＜ 0.05 ). Conclusions MR and CT were superior in detection of the early abnormal changes in evaluating hemophilic arthropathies,however CT could detect more smaller irregular subchondral surface and subchondral cyst formation than MR.

Objective To retrospectively review and compare the clinical results of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) from HLA- matched sibling donors mobilized with different regimens. Methods Seventy-one patients with hematological malignant diseases received allo-PBSCT from HLA-matched sibling donors in our department. Among them, 24 received allografts mobilized with G-CSF (group G), and the remaining (47 cases) were mobilized with G-CSF and GM-CSF (group G+ M). CD34+ subsets and T cell subsets in the allografts were analyzed, and the time of hematopoietic reconstitution and the incidence of graft versus host diseases (GVHD) were compared. The adverse effects on the donors after mobilization were also observed. Results The enough targeted CD34+ cells in all donors were harvested by 1-3 aphereses. Ninety-six h after mobilization, WBC counts of the donors were significantly higher in group G than in group G + M [(49. 6± 19. 5) 109/L vs (25.4 ± 10. 4) 109/L, P＜0. 05]. Analysis of the CD34+ subsets showed that the percentage of cells with the CD34+/CD38- phenotype was significantly higher in group G + M than in group G [(37. 7 ± 5. 7) % vs (31.4 ± 4. 5) %, P＜0. 05]. There was no significant difference in T cells and subsets of grafts. There was no significant difference in the number of total CD34+ cells and CD34+ CD38- cells, and infusion of T cells between two groups. The days required for the recovery of neutrophils and platelets was inversely correlated with the infused CD34+ and CD34+ /CD38- cell number. There was no significant difference in incidence of acute and chronic GVHD between two recipient groups. Seventeen cases and 10 eases among 71 eases died of relapses of primarydiseases, and complications of transplantation such as severe GVHD and infections respectively.Fourteen cases in group G (58.3 %) and 31 cases in group G+ M (66.0 %) survived. The most common adverse events in the donors were bone pain and fever, which mostly occurred 36 h after mobilization and could be relieved by non-steroidal anti-inflammatory drugs. Conclusion Two mobilization regimens showed equivalent clinical results. But the combined regimen of G-CSF and GM-CSF demonstrated a significantly greater mobilization of cells with the CD34+/CD38- phenotype.Meanwhile in allogeneic PBSCT, a greater number of total CD34+ cells and CD34+ CD38- cells infused may be associated with faster hematopoietic reconstitution of recipients.

Objective To evaluate the efficacy and toxicities of gemcitabine plus oxaliplatin with R-GemOx or without (GemOx) rituximab regimen in the treatment of relapsed or refractory diffuse large B-cell lymphoma in elderly patients.Methods A total of 39 patients with relapsed or refractory diffuse large B-cell lymphoma received R-GemOx or GemOx chemotherapy.There were 16 patients in R-GemOx and 23 patients in GemOx group.Patients in both groups received gemcitabine 1000 mg/m2,d1,at land 8 day and oxaliplatin 130 mg/m2,d1 at lday.Patients in R-GemOx additionally received rituximab 375 mg/m2.Every 21-28 days was 1 cycle.The toxicities were evaluated after 1 cycle of chemotherapy.The efficacy was evaluated after 2 cycles of chemotherapy.Results In R-GemOx group,the total response rate was 62.5％,and the clinical benefit rate was 87.5％.In GemOx group,the total response rate was 47.8％,and the clinical benefit rate was 73.9％ There was no significant differences between the two groups.There was a significant difference in the median time-to-progression (TTP) between R-GemOx group (6.4 months) and GemOx group (5.0 months) (P ＜ 0.05).The major toxicities were marrow suppression and gastrointestinal reaction,which had no significant differences between the two groups.Conclusions R-GemOx and GemOx regimen are effective and safe for the elderly patients with relapsed or refractory diffuse large B-cell lymphoma(DLBCL).But the patients with relapsed/refractory DLBCL treated with R-GemOx had a longer median time-to-progression than with GemOx regimen.

To investigate the effects and mechanisms of miR?144 on proliferation, apoptosis and cisplatin ( DDP ) resistance of neuroblastoma cells. Methods Real?time fluorescence quantitative PCR ( RT?qPCR ) was used to detect the mRNA expressions of miR?144 and MYCN in neuroblastoma cell lines, including SH?SY5Y and SK?N?SH, and human umbilical vein endothelial cells HUVEC. The miR?negative control, miR?144 mimics, si?negative control, si?MYCN, miR?144 mimics and pcDNA, miR?144 mimics and pcDNA?MYCN co?transfected SH?SY5Y cells were described as miR?NC, miR?144, si?NC, si?MYCN, miR?144+pcDNA and miR?144+pcDNA?MYCN group, respectively. The half maximal inhibitory concentration ( IC50 ) and cell proliferation were detected by 3?( 4, 5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. The protein expressions of MYCN, p21, cyclin D1, Bax, Bcl?2 were analyzed by western blot.Cell apoptosis was detected by flow cytometry.The cell fluorescence activity was detected by double luciferase reporter gene assay.Results Compared with HUVEC cells, the expressions of miR?144 in neuroblastoma cells SH?SY5Y and SK?N?SH significantly decreased, while the mRNA and protein expression of MYCN significantly increased. The IC50 of DDP was 9.16 μg／ml in SH?SY5Y cells. The absorbance value in 490nm ( A490 value) of miR?144 group was 0.30 ± 0.03, significantly lower than 0.46±0.03 of miR?NC group. The cell apoptotic rate of miR?144 group was 26.94%± 2.01%, significantly higher than 9.68%±0.52% of miR?NC group. The IC50 value of DDP in miR?144 group was 2.95±0.26, significantly lower than 9.23±0.61 of miR?NC group. The expressions of p21, cyclin D1, Bax, Bcl?2 in miR?NC and miR?144 group were 2.67±0.19, 0.41±0.04, 2.12±0.21, 0.18±0.01 and 1.01± 0.07, 1.00 ± 0.06, 1.00 ± 0.05, 1.00 ± 0.06, respectively, with statistical significance ( all P＜0.05). Knockdown of MYCN showed the similar effects with those of miR?144 overexpression in SH?SYSY cells. MiR?144 significantly inhibited the fluorescence activity of ectopic MYCN expressing cells and negatively regulated the expression of MYCN. Overexpression of MYCN can reverse the effects of miR?144 on proliferation inhibition, apoptosis promotion and sensitization of SH?SY5Y cells to DDP. Conclusion MiR?144 inhibits proliferation, promotes apoptosis and enhances the sensitivity of neuroblastoma cells to DDP through targeting MYCN, which provides a potential treatment for neuroblastoma.

Objective To investigate the inhibitory effect of programmed cell death factor 4 (PDCD4) on arsenic trioxide (As2O3)?induced cell growth and nuclear factor kappa B (NF?κB) signaling pathway in neuroblastoma. Methods The PDCD4 overexpression vector was transfected into neuroblastoma cells and detected by fluorescence quantitative PCR and Western blot. As2O3 was used to treat PDCD4 overexpressing neuroblastoma cells. MTT assay was used to measure the proliferation. Colony formation assay was used to determine the cell clone forming ability. Apoptosis was measured by flow cytometry. Western blot was used to detect the expression of NF?κB p65 and cleaved caspase?3 protein in cells. Results The transfection of PDCD4 overexpression vector significantly increased the expression level of PDCD4 in neuroblastoma cells. The cell survival rates of the control group, PDCD4 group, As2O3 group and As2O3+PDCD4 group were 100%, (72.14± 5.20)%, ( 62.58± 3.14)% and ( 40.87 ± 2.47)%, respectively. The colony formation rates in these four groups were (91.25±8.36)%, (65.32±7.14)%, (57.23±5.28)% and (37.14± 3.64)%, respectively. In addition, the cell apoptotic rates of these four groups were ( 3.57 ± 0.24)%, (28.64±3.20)%, (36.41±4.58)% and (49.65±5.27)%, respectively.Therefore, overexpression of PDCD4 in the absence or presence of As2O3 inhibited cell proliferation and clone formation ability, while promoted apoptosis. Furthermore, the expression levels of cleaved caspase?3 in the control group, PDCD4 group, As2O3 group and As2O3+PDCD4 group were 0.21±0.03, 0.30± 0.02, 0.43± 0.05 and 0.57± 0.06, respectively. And the expression levels of NF?κB p65 protein were 0.68±0.04, 0.52±0.03, 0.43±0.04, and 0.32±0.02, respectively. Compared with the control group, the expression levels of NF?κB p65 protein in PDCD4 group, As2O3 group and As2O3+PDCD4 group were significantly decreased (P＜0.05), whereas the expression level of cleaved Caspase?3 protein was significantly increased (P＜0.05). The changes in As2O3+PDCD4 group were more significant than those in PDCD4 group and As2O3 groups ( both P＜0.05 ). Conclusion PDCD4 enhances the inhibitory effect of As2O3 on the growth and NF?κB signaling pathway in neuroblastoma cells.

To investigate the effects and mechanisms of miR?144 on proliferation, apoptosis and cisplatin ( DDP ) resistance of neuroblastoma cells. Methods Real?time fluorescence quantitative PCR ( RT?qPCR ) was used to detect the mRNA expressions of miR?144 and MYCN in neuroblastoma cell lines, including SH?SY5Y and SK?N?SH, and human umbilical vein endothelial cells HUVEC. The miR?negative control, miR?144 mimics, si?negative control, si?MYCN, miR?144 mimics and pcDNA, miR?144 mimics and pcDNA?MYCN co?transfected SH?SY5Y cells were described as miR?NC, miR?144, si?NC, si?MYCN, miR?144+pcDNA and miR?144+pcDNA?MYCN group, respectively. The half maximal inhibitory concentration ( IC50 ) and cell proliferation were detected by 3?( 4, 5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. The protein expressions of MYCN, p21, cyclin D1, Bax, Bcl?2 were analyzed by western blot.Cell apoptosis was detected by flow cytometry.The cell fluorescence activity was detected by double luciferase reporter gene assay.Results Compared with HUVEC cells, the expressions of miR?144 in neuroblastoma cells SH?SY5Y and SK?N?SH significantly decreased, while the mRNA and protein expression of MYCN significantly increased. The IC50 of DDP was 9.16 μg／ml in SH?SY5Y cells. The absorbance value in 490nm ( A490 value) of miR?144 group was 0.30 ± 0.03, significantly lower than 0.46±0.03 of miR?NC group. The cell apoptotic rate of miR?144 group was 26.94%± 2.01%, significantly higher than 9.68%±0.52% of miR?NC group. The IC50 value of DDP in miR?144 group was 2.95±0.26, significantly lower than 9.23±0.61 of miR?NC group. The expressions of p21, cyclin D1, Bax, Bcl?2 in miR?NC and miR?144 group were 2.67±0.19, 0.41±0.04, 2.12±0.21, 0.18±0.01 and 1.01± 0.07, 1.00 ± 0.06, 1.00 ± 0.05, 1.00 ± 0.06, respectively, with statistical significance ( all P＜0.05). Knockdown of MYCN showed the similar effects with those of miR?144 overexpression in SH?SYSY cells. MiR?144 significantly inhibited the fluorescence activity of ectopic MYCN expressing cells and negatively regulated the expression of MYCN. Overexpression of MYCN can reverse the effects of miR?144 on proliferation inhibition, apoptosis promotion and sensitization of SH?SY5Y cells to DDP. Conclusion MiR?144 inhibits proliferation, promotes apoptosis and enhances the sensitivity of neuroblastoma cells to DDP through targeting MYCN, which provides a potential treatment for neuroblastoma.

Objective To investigate the inhibitory effect of programmed cell death factor 4 (PDCD4) on arsenic trioxide (As2O3)?induced cell growth and nuclear factor kappa B (NF?κB) signaling pathway in neuroblastoma. Methods The PDCD4 overexpression vector was transfected into neuroblastoma cells and detected by fluorescence quantitative PCR and Western blot. As2O3 was used to treat PDCD4 overexpressing neuroblastoma cells. MTT assay was used to measure the proliferation. Colony formation assay was used to determine the cell clone forming ability. Apoptosis was measured by flow cytometry. Western blot was used to detect the expression of NF?κB p65 and cleaved caspase?3 protein in cells. Results The transfection of PDCD4 overexpression vector significantly increased the expression level of PDCD4 in neuroblastoma cells. The cell survival rates of the control group, PDCD4 group, As2O3 group and As2O3+PDCD4 group were 100%, (72.14± 5.20)%, ( 62.58± 3.14)% and ( 40.87 ± 2.47)%, respectively. The colony formation rates in these four groups were (91.25±8.36)%, (65.32±7.14)%, (57.23±5.28)% and (37.14± 3.64)%, respectively. In addition, the cell apoptotic rates of these four groups were ( 3.57 ± 0.24)%, (28.64±3.20)%, (36.41±4.58)% and (49.65±5.27)%, respectively.Therefore, overexpression of PDCD4 in the absence or presence of As2O3 inhibited cell proliferation and clone formation ability, while promoted apoptosis. Furthermore, the expression levels of cleaved caspase?3 in the control group, PDCD4 group, As2O3 group and As2O3+PDCD4 group were 0.21±0.03, 0.30± 0.02, 0.43± 0.05 and 0.57± 0.06, respectively. And the expression levels of NF?κB p65 protein were 0.68±0.04, 0.52±0.03, 0.43±0.04, and 0.32±0.02, respectively. Compared with the control group, the expression levels of NF?κB p65 protein in PDCD4 group, As2O3 group and As2O3+PDCD4 group were significantly decreased (P＜0.05), whereas the expression level of cleaved Caspase?3 protein was significantly increased (P＜0.05). The changes in As2O3+PDCD4 group were more significant than those in PDCD4 group and As2O3 groups ( both P＜0.05 ). Conclusion PDCD4 enhances the inhibitory effect of As2O3 on the growth and NF?κB signaling pathway in neuroblastoma cells.

Objective:To explore the correlation between the expression of signaling lymphocyte activation molecule family 6 (SLAMF6) on peripheral blood CD8 +T cells and perforin and granzyme B and the clinical significance in patients with newly diagnosed severe aplastic anemia(SAA). Methods:The indicators of blood routine and bone marrow and peripheral blood samples of 32 newly diagnosed SAA patients admitted to Henan Provincial People′s Hospital from January 2016 to June 2019 were collected for retrospective analysis. Flow cytometry was used to detect the expression of SLAMF6, perforin and granzyme B on samples CD8 +T cell before therapy and 6 months after therapy (11 cases received transplantation, 21 cases received immunosuppressive therapy [IST]). Spearman correlation analysis was performed to determine the association between clinical indicators and laboratory test results. The expression of SLAMF6, perforin and granzyme B was also detected in 10 healthy people (normal group) and 13 myelodysplastic syndromes/paroxysmal nocturnal hemoglobinuria (MDS/PNH) patients (MDS/PNH group). Results:(1) At diagnosis: the expression of SLAMF6 was significantly lower in the SAA group than that in the normal group and the MDS/PNH group ([56.40±6.37]% vs [84.34±5.81]% and [82.24±4.98]% (both P<0.001]). The expression of perforin was significantly higher in the SAA group (32.73±8.46) than that in the normal control group (23.75%±5.10%), and the MDS/PNH group (26.12%±5.53%) (both P<0.05). The expression of granzyme B was also significantly higher in the SAA group (36.23%±7.94%) than that in the normal control group (21.67%±5.05%) and the MDS/PNH group (21.79%±5.10%) (both P<0.001). The expression of SLAMF6 was positively correlated with the hemoglobin ( r=0.804), and reticulocyte absolute values ( r=0.656) in peripheral blood, percentage of granulocytes ( r=0.643) and erythrocytes ( r=0.622) in bone marrow of SAA patients (all P<0.05). Expression of SLAMF6 was negatively correlated with perforin ( r=-0.792) and granzyme B ( r=-0.908) on CD8 +T cells in patients with SAA (both P<0.001). (2) After treatment: the expression of SLAMF6 in peripheral blood CD8 +T cells of 30 surviving patients was higher than pre-treatment ([79.19±12.69]% vs [56.40±6.37]%, P<0.001). The expressions of perforin and granzyme B were lower than pre-treatment level (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 11 transplanted patients was higher than before transplantation ([86.54±3.75]% vs [56.40±7.35]%, P<0.001). The expressions of perforin and granzyme B were lower than before transplantation (both P<0.05). The expression of SLAMF6 on CD8 +T cells in 12 IST-respond patients was higher than that before treatment, while the perforin and granzyme B levels were lower than pre-treatment (all P<0.05). The post-treatment expressions of SLAMF6, perforin and granzyme B were similar as before treatment levels in 7 IST-unrespond patients (all P>0.05). Conclusion:SLAMF6 is significantly down-regulated on CD8 +T cells in newly diagnosed SAA, negatively correlated with the effective factors of CD8 +T cells, which might participate in the immune regulatory of CD8 +T cells as a negative regulatory factor in patients with SAA. The SLAMF6 is significantly up-regulated after hematopoietic recovery, while there is no significant change in treatment-unrespond patients, which could thus serve as an useful diagnostic and therapeutic index of patients with SAA.

Objective: To explore the clinical characteristics and prognostic factors of the patients with non-Hodgkin’s Lymphoma (NHL) complicated with HBV infection, so as to provide a basis for clinical accurate diagnosis and prognosis evaluation. Methods: The data of 313 newly diagnosed NHL patients from August 2012 to July 2016 were collected. The HBV serological markers were detected by ELISA, and HBV DNA was quantified by full automatic microparticle chemiluminescence immunoassay (≥1×10⁵ copies/ml as high copy group, 1×10³-<1×10⁵ copies/ml as low copy group). The relationship between HBV infection and prognosis was analyzed combined with the clinical features of the patients, and the HBV detection rate was compared with that of the common population (from the national HBV sero epidemiological data). Results: ①The positive rate of HBsAg in NHL patients was 12.5% (39/313), which was higher than 7.2% in the general population (χ²=14.596, P<0.001). HBV infection in the past (HBsAg negative but HBcAb positive) in 114 cases (36.4%), the incidence was slightly higher than that in the general population (34.1%). ②Compared HBsAg positive group with the negative group, the proportion of B cell type (87.2% vs 70.3%, P=0.027), Ann Arbor stage Ⅲ-Ⅳ(69.2% vs 34.6%, P<0.001), IPI score 3-5 (74.4% vs 50%, P=0.004), LDH level (79.5% vs 47.8%, P<0.001) and liver involvement (45.5% vs 31.7%, P=0.006) were all higher. The difference was statistically significant. ③Compared the HBV infected group (114 cases) with the non-infected group (160 cases), the difference had statistical significance in the proportion of Ann Arbor stage Ⅲ-Ⅳ (P=0.023) and IPI score 3-5 scores P=0.035). ④Compared HBV DNA positive group (30 cases) with negative group (71 cases), Ann Arbor stage Ⅲ-Ⅳ (P=0.011), IPI score 3-5 score (P=0.030), LDH level (P=0.025) and liver involvement (P<0.001) in the proportion of patients had statistical significance. The positive patients were divided into HBV DNA high and low copy groups with 1×10⁵ copies of /ml as the boundary. The results showed that there was no statistical difference between the two groups (P>0.05). Conclusions The HBV infection rate of NHL patients is significantly higher than that of the general population, and HBV infection is more closely related to B cell type NHL. Patients with HBV infection and HBV DNA positive had late Ann Arbor stage, high IPI score, high LDH level and liver involvement, and the prognosis is poor.

Objective: To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis. Methods: This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45⁺/CD45^- groups. Results: ①The CD45⁺ group was 33 cases (25.38%) , and CD45^- group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45⁺ and CD45^-group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45⁺ and CD45^- group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45⁺ group and CD45^- group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ²=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ²=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 ⁺ group and CD45^- group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ²=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ²=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 μmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors. Conclusion CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45⁺ MM patients.