Objective:To evaluate the value of enhanced magnetic resonance imaging (MRI) on the assessment of synovial hyperplasia and International Prophylaxis Study Group (IPSG) score of hemophilic arthropathy (HA).Methods:This was a retrospective case series study. Briefly, 54 joints of 46 male patients with hemophilia type A and diagnosed with HA in Henan Provincial People′s Hospital from August 2016 to September 2017 were selected. Plain and enhanced MRI were performed at the same time. The IPSG score of synovial hyperplasia and the total joint before and after enhancement were calculated, and the enhancement rate of the synovium and muscles at the same level were also calculated. The differences in synovial hyperplasia and joint total IPSG scores before and after enhancement were compared by paired rank sum test. The correlation between enhancement joint total IPSG score, synovial IPSG score, synovial enhancement rate, total joint bleeding number, course of disease, and Hemophilia Joint Health Score (HJHS) score were analyzed by Spearman′s correlation analysis.Results:Enhanced MRI could more clearly show synovial hyperplasia and ensure better accuracy of joint total IPSG score ( Z=-2.24, P=0.025). The enhancement extent of synovial hyperplasia was higher than that of the same level muscle. There was no correlation between synovial enhancement rate and total number of joint bleeding, course of disease, and HJHS score. After enhancement, the joint total IPSG score was highly positively correlated with the total number of joint bleeding and the disease course ( r=0.96, 0.84, P<0.001) and moderately positively correlated with the HJHS score ( r=0.58, P<0.001). The enhanced synovial IPSG score showed a low positive correlation with the total number of joint bleeding and the disease course ( r=0.37, 0.36, P=0.006, 0.008), but no correlation with HJHS score. Conclusion:Enhanced MRI can provide accurate imaging of synovial hyperplasia of HA and make joint IPSG score more accurate.

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 reveal the related factors of inhibitors and differences ofhemorrhage and joint disease before and after the production of inhibitors in children with hemophilia A (HA) . Methods: Retrospective analyses of the clinical data of 381 children with HA under the age of 16 registered in the Registration Management Center of Hemophilia in Henan Provincial from January 2015 to August 2018. Results: A total of the 381 children were enrolled with 116 (30.4%) mild, 196 (51.4%) moderate, and 69 (18.1%) severe cases; 54 patients (14.2%) had inhibitors, including 22 high and 32 low titer inhibitors. Positive family history was positively associated with inhibitors[P<0.001, OR=3.299 (95%CI 1.743-5.983) ], and high-intensity exposure was associated with inhibitors[P=0.002, OR=2.587 (95%CI 1.414-4.731) ]. High-intensity exposure was associated with high titer inhibitor production[P=0.001, OR=8.689 (95%CI 2.464-30.638) ], and high-intensity exposure increased the risk of high titer inhibitors in HA patients. After inhibitors occurred in 54 patients with HA, the rates of overall joint annual bleeding (z=-3.440, P=0.001) and traumatic annual bleeding (z=-2.232, P=0.026) increased, but the rates of the annual joint bleeding (z=-1.342, P=0.180) and spontaneous annual bleeding (z=-1.414, P=0.157) remained to be not statistically significant. The joint ultrasound score did not change significantly after the inhibitor information (z=-0.632, P=0.527) . Conclusions Positive family history and high-intensity exposure could increase the risk of F Ⅷ inhibitors in HA patients, and high-intensity exposure increased the risk of high titer inhibitors. The rates of the overall joint annual bleeding and traumatic annual bleeding increased after the inhibitor information.

Objective:To evaluate the effect of imatinib on growth impairment in children with chronic myeloid leukemia (CML-CP) in the chronic phase.Methods:From July 2018 to July 2019, questionnaires were distributed to CML children aged <18 years at the time of diagnosis who were receiving imatinib for at least 3 months or to their parents in China. The height-for-age standard deviation score (HtSDS) and the difference of standard deviation integral (△HtSDS) were used to explore the change in height with imatinib therapy.Results:The data of 238 respondents were included; 138 (58.0% ) respondents were men. The median age at the first diagnosis of CML was 11.0 years (range, 1.4-17.9 years) , and 93 (39.0% ) respondents were at the prepuberty stage. At the time of completing the questionnaires, the median age was 15.0 years (range, 2.0-34.0 years) . The median duration of imatinib therapy was 28 months (range, 3-213 months) . Among all the respondents, the mean HtSDS when completing the questionnaires (-0.063±1.361) was significantly lower than that at the time of starting imatinib treatment (0.391±1.244) ( P<0.001) . Total 71.0% respondents showed growth impairment that was more common in those starting imatinib therapy at prepubertal age than in those starting at pubertal age. Multivariate analysis showed that younger at the start of imatinib therapy ( P<0.001) and longer duration of imatinib therapy ( P<0.001) were significantly associated with severe growth impairment on imatinib therapy. Conclusions:Imatinib induced growth impairment in children with CML-CP. Younger the age of initiation and longer the duration of imatinib therapy, more obvious the effect of imatinib on growth impairment.

Objective: 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 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.

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.