The Genetic and Prognostic Characteristics of AML-MRC Patients / 中国实验血液学杂志

OBJECTIVE@#To investigate the genetic and prognostic characteristics of acute myeloid leukemia with myelodysplasia-related changes (AML-MRC) patients.@*METHODS@#There were 230 non-M3 AML patients treated in Ningbo First Hospital enrolled, among which 58 patients were newly diagnosed AML-MRC, the patients were followed up and SPSS 25.0 was used to statistically analyze.@*RESULTS@#There were 49 patients performed genetic testing, 29 patients (59.2%) showed chromosomal abnormalities, including 7q- 8 cases (16.3%), 5q- 6 cases (12.2%), 5 cases (10.2%) of 17p abnormalities, 13 cases (26.5%) of highly abnormal complex karyotypes (CK) (≥5 unrelated chromosomal abnormalities), CK contained chromosomal abnormalities such as +8, 5q-, and 12 cases (24.5%) of monosomal karyotypes (MK). Genetic testing was performed in 37 patients, and 24 (64.9%) patients showed genetic mutations, among which ASXL1 mutation was the most common (8 cases, 21.6%), followed by TET2 mutation in 6 cases (16.2%). Kaplan-Meier analysis showed that AML-MRC patients with high CK (P=0.012), 5q- abnormalities (P=0.038), and TP53 mutations (P=0.008) had poor overall survival.@*CONCLUSION@#AML-MRC has unique genetic characteristics, and high CK, 5q- and TP53 mutations are poor prognostic factors.

Treatment of Chronic Aplastic Anemia with Chinese Patent Medicine Pai-Neng-Da Capsule () for Replacing Androgen Partially: A Clinical Multi-Center Study / 中国结合医学杂志

OBJECTIVE@#To evaluate the efficacy and safety of Pai-Neng-Da Capsule (, panaxadiol saponins component, PNDC) in combination with the cyclosporine and androgen for patients with chronic aplastic anemia (CAA).@*METHODS@#A total of 79 CAA patients was randomly divided into 2 groups by a random number table, including PCA group [43 cases, orally PNDC 320 mg/d plus cyclosporine 5 mg/(kg·d) plus andriol 80 mg/d] and CA group [36 cases, orally cyclosporine 5 mg/(kg·d) plus andriol 160 mg/d]. All patients were treated and followed-up for 6 treatment courses over 24 weeks. The complete blood counts, score of Chinese medical (CM) symptoms were assessed and urine routine, electrocardiogram, hepatic and renal function were observed for safety evaluation. Female masculinization rating scale was established according to the actual clinical manifestations to evaluate the accurate degree of masculinization in female CAA patients treated by andriol.@*RESULTS@#The effective rates were 88.1% (37/42) in the PCA group and 77.8% (28/36) in the CA group based on the standard for the therapeutic efficacy evaluation of hematopathy. There was no significant difference in the white blood cell (WBC) counts, platelet counts and hemoglobin concentration of peripheral blood between two groups after 6 months treatment. The masculinization score of female patient in the PCA group was significantly lower than the CA group (P<0.05). The mild abdominal distention was observed in 1 cases in the PCA group. In CA group, the abnormalities in the hepatic function developed in 2 cases and the renal disfunction was found in 1 case.@*CONCLUSION@#The PNDC possesses certain curative effects in the treatment of CAA without obvious side-effects and can partially replace andriol thereby to reduce the degree of masculinization [Registried at Chinese Clinical Trial Registry (ChicTR1900028153)].

Effect of applying sitting posture corrector on improving reading and writing posture of elementary school students / 上海预防医学

Objective To evaluate the effect of applying sitting posture corrector on improving reading and writing posture of elementary school students， and provide scientific evidence for prevention and control of myopia in children and adolescents. Methods One elementary school each in urban and suburban areas of Shanghai was selected using a convenience sampling strategy. Furthermore， two classes each in Grade 3 and 4 were selected as the intervention group （282 students were included in the study）， and the other two classes each in Grades 3 and 4 were selected as the control group （294 students were included in the study）. Students in the intervention group used the sitting posture corrector in the classrooms for 4 months （from September 2020 to January 2021）， while those in the control group did not use the sitting posture corrector. Relevant data were collected before and after the intervention through a self-administered questionnaire and visual examination. Statistical analysis was performed using chi-square test and generalized estimating equation. Results Before the intervention， 13.5% （38/282） of students in the intervention group and 12.2% （36/294） in the control group had good reading and writing posture （ χ 2 = 0.195， P >0.659）. After the intervention， 18.4% （52/282） of students in the intervention group had good reading and writing posture， which was higher than that （11.2%， 33/294） in the control group （ χ 2=5.957， P =0.015）. Before and after the intervention， there was no significant differences in the prevalence of myopia between students in the intervention and control groups （all P >0.05）. Generalized estimating equation analysis showed that students in the intervention group were 1.502 times more likely to have good reading and writing posture than those in the control group after the intervention （ P =0.043）. Conclusion Applying sitting posture corrector in schools could improve students' reading and writing posture.

Effect of applying sitting posture corrector on improving reading and writing posture of elementary school students / 上海预防医学

Objective To evaluate the effect of applying sitting posture corrector on improving reading and writing posture of elementary school students， and provide scientific evidence for prevention and control of myopia in children and adolescents. Methods One elementary school each in urban and suburban areas of Shanghai was selected using a convenience sampling strategy. Furthermore， two classes each in Grade 3 and 4 were selected as the intervention group （282 students were included in the study）， and the other two classes each in Grades 3 and 4 were selected as the control group （294 students were included in the study）. Students in the intervention group used the sitting posture corrector in the classrooms for 4 months （from September 2020 to January 2021）， while those in the control group did not use the sitting posture corrector. Relevant data were collected before and after the intervention through a self-administered questionnaire and visual examination. Statistical analysis was performed using chi-square test and generalized estimating equation. Results Before the intervention， 13.5% （38/282） of students in the intervention group and 12.2% （36/294） in the control group had good reading and writing posture （ χ 2 = 0.195， P >0.659）. After the intervention， 18.4% （52/282） of students in the intervention group had good reading and writing posture， which was higher than that （11.2%， 33/294） in the control group （ χ 2=5.957， P =0.015）. Before and after the intervention， there was no significant differences in the prevalence of myopia between students in the intervention and control groups （all P >0.05）. Generalized estimating equation analysis showed that students in the intervention group were 1.502 times more likely to have good reading and writing posture than those in the control group after the intervention （ P =0.043）. Conclusion Applying sitting posture corrector in schools could improve students' reading and writing posture.

Clinical Features of Pregnant Women with Thalassemia in Non Endemic Area / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical features of pregnant women with thalassemia in non endemic area, and to prevent the births of babies with intermedia or major thalassemia.@*METHODS@#Two hundred and thirty-five pregnants women with thalassemia diagnosed from March 2015 to April 2016 in our hospital were enrolled and retrospectively analysed. The blood routine and hemoglobin electrophoresis were performed respectively by XN-9000 automatic blood cell analyzer and HYDRASYS hemoglobin electrophoresis apparatus. The three commonest deletion of α-thalassemia, the three non-deletion α-thalassemia and 21 known β-thalassemia mutation were all detected by fluorescence melting curve analysis.@*RESULTS@#Among 235 pregnant women of thalassemia, the majority were β-thalassemia, which were followed by α-thalassemia and composite thalassemia. Most pregnant women showed a mild anemia, and suffered from microcytic anemia, but less suffered from iron deficiency anemia. The ratio of second-child pregnant women was increased, and the ratio was close to one third both in α-thalassemia and β-thalassemia patients, and 75% patients were composite thalassemia. HbF was found to be more in native pregnant women with β-thalassemia. Hemoglobin isomer was easy to found in the pregnant with α-thalassemia, and they were all non native. The genotype of --@*CONCLUSION@#More pregnant women with thalassemia are founded to be in non endemic area, and shows their own unique clinical features. It is certainly to detect thalassemia mutation in their spouse and their babies, to prevent the births of babies with intermedia or major thalassemia.

Influence of nimodipine on neurological function ,serum levels of AQP4 and MIF in aged patients with hypertensive intracerebral hemorrhage / 心血管康复医学杂志

Objective :To study influence of nimodipine on neurological function and serum levels of aquaporin (AQP) 4 and macrophage migration‐inhibitory factor (MIF) in aged patients with hypertensive intracerebral hemorrhage (HICH).Methods :A total of 120 aged HICH patients treated in our hospital from Jan 2014 to Jan 2017 were ran‐domly and equally divided into routine treatment group (received puncture removal of hematoma + routine treat‐ment) and nimodipine group (received nimodipine based on routine treatment group ) ,both groups were treated for three weeks.United States National Institutes of Health Stroke Score (NIHSS) score ,brain edema volume ,serum levels of AQP4 ,MIF ,hsCRP and tumor necrosis factor (TNF)‐α ,therapeutic effect and incidence of adverse reac‐tion were measured and compared between two groups before and after treatment .Results :Total effective rate of nimodipine group was significantly higher than that of routine treatment group (88.33% vs .70.00%) , P＝0.013. Compared with routine treatment group after treatment ,there were significant reductions in NIHSS score [ (17.39 ± 3. 08) scores vs .(12. 26 ± 3.74) scores] ,brain edema volume [ (11. 84 ± 2.31) ml vs .(6.68 ± 1.93) ml] ,serum levels of AQP4 [ (2. 79 ± 0. 64) ng／ml vs .(1. 84 ± 0. 52) ng／ml] ,MIF [ (55.39 ± 7. 65) ng／L vs.(43.25 ± 5. 81) ng／L] ,hsCRP [ (18.83 ± 5. 17) mg／L vs.(12. 53 ± 3.87) mg／L] and TNF‐α [ (8. 42 ± 1.37) ng／L vs.(5. 78 ± 1.96) ng／L ] in nimodipine group , P＝0.001 all.There were no severe adverse reactions and no significant differ‐ence in incidence rate of adverse reactions between two groups , P＝0.436. Conclusion :Nimodipine can significantly improve therapeutic effect ,neurological function ,reduce serum AQP4 and MIF levels ,relieve brain edema and in‐flammation in aged HICH patients .

Effect of Chromosomal Karyotype on the Prognosis of Patients with Acute Promyelocytic Leukemia in Condition of the Maintenance Treatment Based on Arsenic Trioxide / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of chromosomal karyotype on the prognosis of patients with acute promyelocytic leukemia (APL) in condition of the maintenance treatment based on arsenic trioxide.@*METHODS@#The patients with acute promyelocytic leukemia for last 12 years in our hospital were retrospectively collected. The patients mainly treated with arsenic trioxide in maintenance protocol were selected and followed up. All the patients were divided into 3 groups according to cytogenetic data: single t (15; 17) group, t (15; 17) with additional chromosomal abnormality (ACA) group, and normal karyotype group. Then, the prognostic significance of ACAs and complex karyotype were investigated in APL patients.@*RESULTS@#There were 57 cases in the single t (15; 17) group, in which 8 cases died in the first month after induction treatment with early mortality rate of 14%. There were 21 patients in t (15; 17) with ACA group, in which 4 cases died in the first month with early mortality rate of 19%. There were 15 cases in normal chromosome group, in which 5 cases died in the first month with the early mortality rate of 33.3%. There was no statistical difference in the early mortality among 3 groups. All the remaining 76 patients achieved complete hematological remission. These patients were followed up. The median follow-up time was 43.9 months. Among them, only 2 patients in single t (15; 17) group and 1 patient in t (15; 17) with ACA group relapsed. No patient relapsed in normal karyotype group. The relapse rate was 3.5% in single t (15; 17) group and 4.2% in t (15; 17) with ACA group, respectively. There was no statistical difference in the overall survival and disease-free survival rates among 3 groups. Further analysis showed that the patients with complex chromosome karyotypes had lower relapse-free survival rates, but overall survival rates were not significantly different in 3 group.@*CONCLUSION@#In general, ACA can not affect the prognosis of patients with acute promyelocytic leukemia in condition of the maintenance treatment based on arsenic trioxide, but the complex chromosomal karyotype may reduce the relapse-free survival rates.

Clinical Significance of Monosomal Karyotype in MDS / 中国实验血液学杂志

OBJECTIVE@#To investigate the frequency, karyotype characteristics and prognosis significance of monosomal karyotype (MK) in newly diagnosed MDS patients.@*METHODS@#The clinical, laboratorial and follow-up data of 202 MDS patients received the chromosome karyotype test in Department of Hematology, Ningbo Hospital of Zhejiang University from 2009 to 2018 were analyzed retrospectively, the monosomal karyotype features, clinical characteristics and their effects on the prognosis of MDS patients also were analyzed.@*RESULTS@#Among 202 cases of MDS, 25 (12.38%) confirmed to be the MK. The abnormality of chromosome 5 (60.00%), 7 (56.00%), 17 (56.00%), 15 (56.00%), 13 (40.00%) and 20（40.00%）were common in monosomal karyotype. MK-MDS (MDS with monosomal karyotype) patients had higher bone marrow blast percentage than MK-MDS (MDS without monosomal karyotype) patients, the median are 6.25% and 3.00% (P＜0.01) respectively, but there were no difference in age, sex, hemoglobin level, white blood cell count, neutrophile granulocyte percentage, platelet count, blood blast percentage, serum ferritin, folic acid and vitaminB12 between MK-MDS and MK-MDS. The overall survival time of MK-MDS and MK-MDS patiens with chromosome 3, 5, 7, 13, 15, 17 abnormalities was significantly shorter than MK-MDS and AK+MK-MDS patients (MDS with abnormal karyotype but without monosomal karyotype) , the MK-MDS patients had a median survival time of 7.33 months, but the median survival time had not been reached in MK-MDS and AK+MK-MDS patients had not been reached by the end of the follow-up, and could not be assessed (P＜0.01).@*CONCLUSION@#The monosomal karyotype is a poor prognosis factor for newly-diagnosed MDS patients. The poor prognosis suggested by monosomal karyotype may be related with the abnormality of 3, 5, 7, 13, 15 and 17 chromosome.

Clinical Characteristics and Karyotype Analysis of Myelodysplastic Syndrome Patients with TP53 mutation / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical characteristics of myelodysplastic syndrome (MDS) with TP53 mutant and the relationship between TP53 mutation and monosomal karyotype in MDS patients.@*METHODS@#The TP53 mutations in 102 patients with de nove MDS were retrospectively analyzed, and the clinical features of the TP53 mutation group and the non-mutation group were compared. The relationship between TP53 mutation and karyotype, especially monosomal karyotype was analyzed.@*RESULTS@#Fifty-two out of the 102 MDS patients were male and 50 were female, the median age was 59.5 (23-83) years old. The mutational frequency of TP53 was 12.7%, which mostly occurred in patients with MDS-EB. As compared with non-mutation group, the hemoglobin level and platelet count were lower (P=0.001, P=0.033), the LDH level and bone marrow blast ratio were higher in TP53 mutation group (P=0.002, P＜0.001), but the statistical difference of alsolute count of neutrophils and levels of serum ferritin and β2-microglobulin between 2 groups was not found. The karyotype abnormality frequency of patients with TP53 mutation was 90.9%, among them 72.7% was monosomal karyotype. The incidence of monosomal karyotype in the TP53 mutation group was very significantly higher than that in the non-mutation group (P＜0.001). MDS with TP53 mutation and monosomal karyotype appeared in the groups with high and very high IPSS-R risk.@*CONCLUSION@#MDS patients with TP53 mutation have unique clinical features and high incidence of monosomal karyotype, and their overall prognosis is poor.

The preliminary analysis of double stent-retriever thrombectomy for acute internal carotid artery occlusion / 中华神经科杂志

Objective To explore the safety and efficacy of Solitaire AB double stents in acute occlusions in bifurcation of cerebral artery (including the ends of internal carotid artery and middle cerebral artery M1 segment).Methods The clinical and imaging data of six cases treated with the double stent retriever technique using the Solitaire AB system in Wuhan No.1 Hospital from January to November 2017 were retrospectively analyzed.And the therapeutic effect and postoperative complications of them were analyzed.Results One patient took the double stents directly,whereas five patients were treated with double stent-retriever thrombectomy after the failure of single stent thrombectomy.All of the six patients achieved recanalization successfully (modified thrombolysis in cerebral infarction (mTICI) 3 in five patients,mTICI 2b in one).All patients had no intracranial hemorrhage immediately after thrombectomy.In the 24 hours,7 days and 2 weeks,the median NIHSS score was 10 (3-17),3 (1-15) and 1 (0-15),respectively.During perioperative period,one patient had asymptomatic cerebral hemorrhage,one died of symptomatic cerebral hemorrhage within 48 hours,and one was complicated with pulmonary infection.Five patients were followed up by outpatient visit,and four patients showed good outcome (modified Rankin Scale score ≤2).Conclusion In the emergency revascularization of acute cerebral artery occlusion at arterial bifurcation,double stent-retriever is better at increasing the efficacy of thrombectomy and is safe compared with single stent mechanical thrombectomy.

Evaluation of Circulating Endometrial Cells as a Biomarker for Endometriosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Circulating endometrial cells (CECs) have been reported to be present in the peripheral blood of women with endometriosis (EM), providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs.</p><p><b>METHODS</b>The participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay.</p><p><b>RESULTS</b>The detection rate of CECs reached 89.5% (17/19) in the EM group, which was significantly higher than that of the control group (15.0% [6/40], P < 0.001) and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage I-II EM. In contrast, a positive CA125 test had limited value in detecting EM (13/19, 68.4%) and detected only one case of Stage I-II EM.</p><p><b>CONCLUSION</b>CECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay.</p>

Effects of Bone Marrow Mesenchymal Stem Cells on the Biological Characteristics of Migrating and Homing in Multiple Myeloma Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To construct a co-culture system for bone marrow mesenchymal stem cells (BMMSC) and multiple myeloma (MM) cells, and to investigate the effects of co-cultured BMMSC on the migrating and homing of multiple myeloma cells.</p><p><b>METHODS</b>The BMMSC from the transgenic mice with green fluorescent protein (GFP) fetal bone were cultured by adherent screening. A co-culture system of BMMSC and MM cell line XG-7 cells was constracted, the proliferation and apoptosis of cells were determined by trypan blue exclusion and Annexin V/PI, respectively, MDC staining was employed to detect the autophagy. The moving direction distribution of molecule in BMMSC and XG-7 cells labeled with PE-CD138 in co-culture process were observed dinamically by confocal microscopy.</p><p><b>RESULTS</b>After co-culture with GFP-BMMSC, the resistance of XG-7 cells to apoptosis and autophagy were enhanced; at the same time, their proliferation increased. Apoptosis rates of XG-7 cells directly and indirectly co-cultured with BMMSC were (6.23 ± 0.12)% and (6.97 ± 0.03)% respectively, which were lower than that of XG-7 cells cultured alone (17.90 ± 1.46)% (P < 0. 01). There was low level of autophagy in XG-7 cells co-cultured with BMMSC. XG-7 cells are highly polarized and contained a specialized membrane domain with specific protein and lipid components to contact with BMMSC under confocal microscope. After methyl-β-cyclodextrin treatment, the molecules were normally enriched in the specialized domain.</p><p><b>CONCLUSION</b>BMMSC can protect XG-7 cells from apoptosis and autophagy, and obviously promote the proliferation of XG-7 cells, and can influence the migrating and homing of multiple myeloma cells.</p>

Supporting Effect of Umbilical Cord Blood-Derived Mesenchymal Stem Cells on CD34+ Cell Proliferation and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the ability of UCB-derived MSCs to support the expansion of HSCs ex vivo and the possible mechanisms involved in this process.</p><p><b>METHODS</b>HSCs from UCB were co-cultured with UCB-derived MSCs for 14 days, and then the total number of HSCs and colony-forming units (CFU) were detected. Cytokines levels of MSCs supernatant were analyzed using ELISA.</p><p><b>RESULTS</b>The proliferation rate of HSCs co-cultured with MSCs was significantly higher than that of cultured HSCs alone (P<0.05). Furthermore, the addition of exogenous cytokines to the culture system significantly increased the proliferation rate of HSCs (P<0.05). MSCs had secretion of many cytokines, including GM-CSF, IL-7, IL-8, IL-11, SCF and SDF-1α.</p><p><b>CONCLUSION</b>UCB-derived MSCs as a feeder layer can be an alternative approach for ex vivo expansion of HSCs, and the cytokines by secreted UCB-MSCs may mediate the supportive role of MSC to HSC proliferation.</p>

Effect of umbilical cord blood mesenchymal stem cells on peripheral blood lymphocyte subsets / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the effect of mesenchymal stem cells (MSCs) on subsets and cytokine secretion of T lymphocytes.</p><p><b>METHODS</b>Umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) were isolated by density gradient and were cultared by amplifying culture. The subsets and cytokine secretion of T lymphocytes were detected by flow cytomety after being co-cultured with UCBMSC.</p><p><b>RESULTS</b>The proliferation of lymphocytes was inhibited. CD4(+)T cell subsets were increased, CD8(+)T cell subsets decreased when co-cultured with UCBMSC; Th1 and Tc1 level significantly reduced, while Th2 and Tc2 level slightly increased.</p><p><b>CONCLUSION</b>The UCBMSC can inhibit the proliferation of lymphocytes, especially CD8(+)T cell subsets. In addition, UCBMSCs can reduce Th1 and Tc1 cells, and increase Th2 and Tc2 cells. UCBMSC may have the clinical application potential for preventing and remedying GVHD.</p>

Mechanism of HLA-G participation in inhibiting lymphocyte proliferation by amniotic mesenchymal stem cells / 中国实验血液学杂志

This study was aimed to investigate the inhibitory mechanism of human amniotic mesenchymal stem cells (HAMSC) on lymphocyte proliferation and to validate the participation of the nonclassic human leukocyte antigen (HLA) class I molecule (HLA-G) in immunosuppressive action of HAMSC. HAMSC were isolated from fetal membranes of human placentas, and were cultured and expanded. The phenotypes of HAMSC were identified by flow cytometry, at same time the HLA-G levels on membrane surface and in cytoplasm were detected by flow cytometry. The soluble HLA-G (sHLA-G) level in HAMSC supernatants was determined by ELISA, MTT assay was used to examine the effect of mixed cultured HAMSC on proliferation of lymphocytes. The results showed that both surface and cytoplasm of HAMSC expressed HLA-G, the average rates of HLA-G expression on surface and in cytoplasm were (16.75 ± 3.871)% and (39.14 ± 4.274)%, respectively. The sHLA-G level in cell culture supernatant was 5.2 ng/ml. After HAMSC and culture supernatants were added in the MLR, the inhibitory rate on lymphocyte proliferation increased obviously, meanwhile the inhibitory rate on lymphocyte proliferation decreased when the HLA-G antibody was added in MLR. It is concluded that the surface and cytoplasm of HAMSC express HAL-G, at same time HAMSC secrete the HLA-G to supernatants of culture. The HLA-G is one of critical factors inhibiting immuno-function of HAMSC. This study contributes to improve the clinical therapeutic trails for using the HAMSC to prevent rejection.

Effects of arsenic trioxide combined with bortezomib on apoptosis of multiple myeloma cell line KM3 and its mechanisms / 中国实验血液学杂志

This study was purposed to investigate the effect of bortezomib (Bor) and arsenic trioxide (As(2)O(3)) combination on multiple myeloma cell line KM3 and its mechanisms. KM3 cells were cultured with different concentration of Bor or As(2)O(3) as well as both for a certain time. The cell proliferation was analysed by MTT assay and the concentration of 50% proliferation inhibition (IC(50)) was calculated. Early apoptosis and late apoptosis of KM3 cells were detected by Annexin-V-FITC Kit, and the change of transmembrane potential was measured by flow cytometry. mRNA of Caspase-3, Bim and Bcl-xL were detected by RT-PCR. The results showed that the proliferation inhibitory rate of KM3 cells treated by Bor plus As(2)O(3) was much higher than that of KM3 cells treated by Bor only for 72 h [ (27.64 ± 0.81)% vs (21.67 ± 2.20)%, P < 0.05]. There were more KM3 cells treated by Bor plus As(2)O(3) in early apoptosis at 48 h and late apoptosis at 72 h than that of KM3 cells treated only by Bor [ (53.20 ± 3.70)% vs (35.40 ± 2.58)%, P < 0.01; (63.96 ± 2.97)% vs (54.08 ± 3.76)%, P < 0.01]. Transmembrane potential (Δψm) of KM3 cells treated by Bor plus As(2)O(3) decreased more at 48 h, as compared with Bor alone. The expression levels of caspase-3 mRNA and Bim mRNA in KM3 cells treated with Bor plus As(2)O(3) were higher than that in KM3 cells treated with Bor alone. But the expression level of Bcl-xL mRNA was lower than that in KM3 cells treated with Bor alone. It is concluded that As(2)O(3) can enhance the apoptosis-inducing effect of Bor on multiple myeloma cell line KM3, which is associated with decreasing the expression of Bcl-xl mRNA and increasing the expression of Caspase-3 and Bim mRNA.

Phenotypic identification and differentiation potential analysis of two kinds of human amniotic cells / 中国实验血液学杂志

The aim of this study was to isolate, cultivate and phenotypically characterize two types of human amnio-tic membrane (HAM)-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells (hAEC) were derived from the embryonic ectoderm, while human amnion mesenchymal cells (hAMC) were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immunofluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers (CD29, CD73, CD44, CD59, CD90, CD105, CD166), but did not express the hematopoietic markers (CD31, CD34, CD45, HLA-DR) and showed the weak expression of costimulatory molecules (CD40, CD40L, CD80, CD86). Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytes potential, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.

Induction of dendritic cells with multidrug resistance from K562/MDR1 cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To induce the differentiation of K562/MDR1 cells into dendritic cells (DC) with multidrug resistance property.</p><p><b>METHODS</b>K562/MDR1 cells and K562 cells were cultured in the presence of GM-CSF and IL-4 to generate DC and matured by TNF-α. On d14 K562/MDR1-DC and K562-DC cells were harvested and the expressions of CD1a, CD83, CD80, CD86, HLA-ABC and HLA-DR were assessed by flow cytometry (FCM). The antigen presentation function of K562/MDR1-DC and K562-DC was determined by allogenic mixed lymphocyte reaction (Allo-MLR). The expression of P-glycoprotein and the intracellular accumulation of daunorubicin (DNR) were detected by FCM. The sensitivity of K562/MDR1-DC and K562-DC cell to vincristine, adriamycin was measured using MTT assay.</p><p><b>RESULTS</b>Both K562/MDR1 and K562 cells were differentiated into dendritic cells in the presence of cytokine cocktails, showing the morphologic and immunophenotypic characteristics of DC. K562/MDR1-DC more markedly enhanced proliferation of allogeneic lymphocytes in MLR than K562-DC. High level expression of P-glycoprotein and efflux of DNR were demonstrated in K562/MDR1-DC. K562/MDR1-DC showed multidrug resistance property, with higher IC(50) to VCR and ADM than that of K562-DCs.</p><p><b>CONCLUSION</b>K562/MDR1 cells can be differentiated into DC with the presence of cytokines, the induced K562/MDR1-DC cells express high level of P-glycoprotein and acquire the multidrug resistance property.</p>

Influence of HLA-G on immunoregulatory function of human amniotic mesenchymal cells / 中国实验血液学杂志

Human leukocyte antigen G (HLA-G), a kind of non-classical major histocompatibility complex class I antigens, can inhibit inflammatory reaction, assist tumor cells to escape from immune surveillance and promote the immunologic tolerance of the graft. HLA-G, expressed and secreted by human amniotic mesenchymal cells (HAMC), suppresses the functions of NK cells, T cells and B cells and modulates the activity of dendritic cells (DC). These findings provide a theoretical basis for illustrating the mechanism of immunosuppression on HAMC. In this article, the recent advances on not only the gene and the molecular structure of HLA-G, but also the possible mechanisms of HLA-G in immunoregulatory function of HAMC, as well as the relation of HLA-G with HAMC, NK, DC, T and B cells are reviewed.

In vitro and in vivo antifungal activity of a triadimenol compound / 第二军医大学学报

Objective: To observe the in vitro and in vivo antifungal activity of a triadimenol compound SL-130. Methods: The M27-A project recommended by the National Committee for Clinical Laboratory Standard (NCCLS) in 1997 was used in the present study. The minimum inhibitory concentrations (MIC80) of SL-130 were determined for candidal and non-candidal strains with different susceptibilities to fluconazole. Agar disk diffusion test, time-kill curves test and mouse survival rate were employed to examine the antifungal activity of SL-130. Results: The results indicated that SL-130 had stronger in vitro antifungal activity than fluconazole, and similar in vivo activity to fluconazole. Conclusion: The structure of SL-130 is quite unique and it has in vitro and in vivo antifungal activity: further study is warranted on the compound.