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【Objective】 To optimize the existing spin-EB method and promote human induced pluripotent stem cells (hiPSCs) differentiate into megakaryocytes (MKs). 【Methods】 In this study, the initial inoculation amount of hiPSCs was increased from 3 500 cells/well to 8 000 cells/well, and the size of EB was increased. By observing the generation time of EB- hematopoietic cells during differentiation, and detecting the proliferation of CD34+ hematopoietic progenitor cells and CD41+ MKs in different stages, it was studied whether the optimized scheme could promote the differentiation of hiPSCs into hematopoietic progenitor cells(HPCs) and MKs. 【Results】 By increasing the initial inoculation amount of hiPSCs and the size of EB, the differentiation of hiPSCs into HPCs and MKs and the cell production efficiency can be promoted. 【Conclusion】 Our research describes an optimized and repeatable differentiation method, which can produce hematopoietic progenitor cells and mature MKs from hiPSCs in a relatively short time with higher yield. It is of great clinical significance and broad scientific research prospect to continuously optimize the culture scheme of hiPSCs differentiation to produce MKs and platelets in vitro, and to promote large-scale platelet generation in vitro in transfusion medicine.
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Objective:To compare the expression of myeloid cell trigger receptor expressed on myoid cell 2 (TREM2) in different brain regions of tyrosine kinase binding protein(TYROBP) knockout mice and wild-type mice at different months of age, and to explore the relationship between TREM2, TYROBP and early onset Alzheimer's disease(EOAD).Methods:Healthy TYROBP gene knockout mice were divided into three groups according to the results of gene sequencing: the homozygous (TYROBP -/-) group, the heterozygous (TYROBP -/+ ) group, and the wild type (WT) group.Western blot and RT-qPCR were used to detect the expression of TREM2 in prefrontal cortex and hippocampus of 2, 4 and 6 month old mice in the three groups and with 10 in each group at each time point. Results:(1) In the prefrontal cortex: Western blot and RT-qPCR results showed that compared with WT mice (2-month-old: (0.993±0.048), (1.654±0.033); 4-month-old: (0.503±0.019), (2.169±0.023); 6-month-old: (0.600±0.036), (1.468±0.057)), the levels of TREM2 protein and mRNA in 2-month-old TYROBP -/+ group ((0.746±0.062), (1.137±0.067)) and TYROBP -/- group ((0.661±0.028), (0.644±0.012)) were decreased.While in 4-month-old and 6-month-old TYROBP -/+ group (4-month-old: (1.140±0.006), (5.483±0.088); 6-month-old: (0.827±0.043), (3.020±0.082)) and TYROBP -/- group (4-month-old: (1.071±0.010), (3.012±0.150); 6-month-old: (0.627±0.026), (1.633±0.027)) were increased, especially in 4-month-old mice and the differences were statistically significant ( F=12.946, 134.445; 725.318, 289.202; 12.172, 202.791; all P<0.05). (2) In the hippocampus: Western blot results showed that compared with WT mice (2-month-old: (1.268±0.036); 4-month-old: (0.813±0.010); 6-month-old: (0.312±0.021)), the level of TREM2 protein in 2-month-old TYROBP -/+ group ((0.804±0.034)) and TYROBP -/- group ((0.534±0.020)) were decreased.While in 4-month-old and 6-month-old TYROBP -/+ group ((0.932±0.011); (0.769±0.031)) and TYROBP -/- group ((0.910±0.014); (0.609±0.018)) were increased, especially in 4-month-old mice and the differences were statistically significant ( F=142.807; 27.884; 94.067; all P<0.05). Conclusion:The expression level of TREM2 decreases in 2-month-old TYROBP gene knockout mice while increases in 4-month-old and 6-month-old TYROBP gene knockout mice.It is presumed that TREM2/TYROBP signal pathway participates in the pathological process of EOAD and plays different roles in different pathological stages of EOAD.
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Objective To study the association between the single nucleotide polymorphism (SNP) of SEPT14 (rs77231105,rs10241628,rs11981883,rs73701167) and sporadic PD in Chinese Han populationin of Southwest Shandong.Methods One hundred and eighty PD patients from Southwest Shandong served as PD patient group and 200 healthy subjects from Southwest Shandong served as control group in this study.The distribution frequencies of alleles and genotypes in SNP of rs77231105,rs10241628,rs11981883 and rs73701167 were compared by PCR and sequencing respectively.Results No significant difference was found in the distribution frequencies of alleles and genotypes in rs77231105,rs10241628,rs11981883 between the two groups (P＞0.05).The distribution frequency of rs73701167 was significantly higher in PD patient group than in control group (31.1％ vs 20.5％,OR=1.75,95％CI=1.261-2.428,P=0.001).Conclusion The SNP of SEPT14 rs77231105,rs10241628 and rs1198188 are not associated with PD,the SNP of rs73701167 is associated with PD in Chinese Han population of Southwest Shandong.The allele C is a risk factor for PD.
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Objective To investigate the expression of ferroptosis-related proteins in the hippocampus in AD mice. Methods AD mice were created by knocking in PSEN1 p.G378E gene in C57BL/6 mice. The expression of ferroptosis-related proteins, glutathione peroxidase 4 (GPX4), cystine-glutamate exchanger (SLC7A11), long-chain fattyacyl-CoA synthetase 4 (ACSL4) and phosphatidylethanolamine-binding protein1 (PEBP1) in the hippocampus were compared among PSEN1 p.G378E-/-、 PSEN1 p.G378E-/+and WT mice. Results The expression levels of GPX4, PEBP1, SLC7A11 and ACSL4 proteins in the hippocampus were significantly increased in PSEN1 p.G378E-/-mice than in either PSEN1 p.G378E-/+or WT mice (P<0.05). Meanwhile, the expression levels of GPX4 and PSEN1 proteins were higher in PSEN1 p.G378E-/+mice than in WT mice (P<0.05). However, there were no significant differences in expression levels of SLC7A11 and ACSL4 proteins between PSEN1 p.G378E-/+mice and WT mice (P>0.05). Conclusion Expression levels of ferroptosis-related proteins are significantly increased in the hippocampus of PSEN1 p.G378E knock-in mice.
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Objective To explore the clinical characteristics and prognostic factors of breast carcinosarcoma.Methods Clinical data of 37 breast carcinosarcoma patients treated at Tianjin Cancer Hospital from 1974 to 2014 was analyzed retrospectively.Kaplan-Meier unvariate analysis was used to calculate the survival rate and compare the survival rates.Multivariate factors for survival were analyzed by COX proportional hazards regression model.Results 37 cases of breast carcinosarcoma were all female.The median age was 55 years.Among them 31 cases complained for painless mass,six cases had breast mass with skin ulceration.21 cases were three-negative breast cancer.Ten cases had axillary lymph node metastasis,ten cases had distant metastasis.The 5-year survival rate was 62.3％.Unvariate analysis showed that axillary lymphnodes metastasis (P =0.045) and therapeutic schedule (P =0.016) significantly influenced the oulcome of the patients.COX multivariate analysis validated that the therapy modality was an independent prognostic factor for breast carcinosarcoma(P =0.041).Conclusions Breast carcinosarcoma is rare and has a poor prognosis.ER,PR and HER-2 expressions in most cases are negative.If there is axillary lymph node metastasis,the prognosis is poor.A reasonable and comprehensive treatment can improve the prognosis.
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Objective To study the clinical characteristics of acute leukemia(AL) patients with cross‐lineage antigen expression . Methods Patients were diagnosed and classified by morphology ,cytochemistry and immunology assay ,and prognostic acting factor were also analyzed .Results According to FAB standards ,acute myeloid leukemia(AML)‐M2 ,acute lymphocytic leukemia (ALL)‐L2 and no‐classified type were common in 320 patients with cross‐lineage antigen expression .The immunophenotype with B and my‐eloid mixed expression was the most common(176 cases) ,followed by cross expression of antigen T and myeloid(131 cases) ,and the co‐expression of B ,T and myeloid antigen was found in only 10 cases .In lymphoid antigenpositive AML(Ly+ AML) ,CD19 anti‐gen was the most common among B lineage ,CD7 was the most common in T lineage .In myeloid antigen positive ALL(My+ ALL) , CD33 was the most common myeloid antigen .Forty‐five cases were with mixed expression of myeloid antigen and CD56 expression . Correlation existed between CD7 and CD34(P< 0 .05) ,CD19 and CD34(P< 0 .05) .There were 9 patients with CD34 ,CD7 and CD19 co‐expression ,7 patients with CD34 ,CD7 and CD56 co‐express .In Ly+ AML patients ,23 cases were with recurrent chromo‐some abnormality ,including 11 cases with t(8 ;21)(q22 ;q22) ,RUNX1‐RUNX1T1 ,3 cases with t(15;17)(q22 ;q11‐12) ,PML/RAR ,6 cases with bone marrow eosinophilia inv(16)(p13 ;q22) ,CBF beta /MYH11 ,and 3 cases with t(9;11)(p22;q23) , MLLT3‐MLL .In My+ ALL ,15 patients were with recurrent chromosome abnormality ,including 9 cases of B‐ALL with t(9;22) (q34 ;q11 .2) ,BCR‐ABL1 ,3 cases of B‐ALL with t(v ;11q23) ,MLL rearrangement ,and 3 cases of T‐ALL with 14q11 .2 .Among the presence of reproducible chromosomal abnormalities in AL ,the antigen expression of mistranslation was still with a certain fea‐ture:Ly+ AML patients often mistranslated CD19 ,CD56 ,CD2 ,and My+ ALL patients often mistranslated CD13 and CD33 .Com‐pared with the lymphoid antigennegative AML(Ly - ALL) group ,CD7+ AML group ,CD19+ AML group and CD56+ AML group had significant difference in survival curves(with P value of 0 .01 ,0 .02 and 0 .02) .There was no significant difference in survival curves between myeloid antigen negative ALL(My -ALL) group and CD13/33+ ALL group(P<0 .05) .CD7 was also positive com‐monly(53 cases) and related with CD34(P<0 .05) .So it significantly influenced the prognosis .If patients were with co‐expression of CD34 ,CD7 and CD19 ,the prognosis could be worse .Conclusion AL with cross‐lineage antigen expression might be a special type and confirmed by immunotype .Furthermore ,expression types of differentiation antigen could be critical for prognosis and sur‐vival .
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Objective To know the condition of cognition of cardiac surgical patients for patient safety.Methods Carried out a semi-structured depth interviews among 8 cardiac surgical patients,and then cleared up the information by phenomenology method.Results Three themes were extracted:patients' consciousness about patient safety,the lack of knowledge of patients in patient safety management,the lack of emotional support of patients in patient safety.Conclusions Some positive counter-measures should be taken by nursing administrators to resolve patients' difficulty and confusion,and then promote patients take part in the course of safety management of patients,ensure patients' safety.
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Objective To investigate the curative effects of oxalic acid Ai Sciplan in assisting psychological therapy for coronary heart disease accompanied with depression. Methods A total of 136 patients of coronary heart disease with de?pression were randomly divided into control group and treatment group. The routine drug treatment was given to control group. Treatment group was given oxalic acid Ai Sciplan which is assisted by the psychological therapy based on the routine drug treatment schedule for six months. HAMD score before treatment, one-week, one-month, 3-month and 6-month after treatment were compared between two groups. Homocysteine (Hcy) and high sensitive C reactive protein (hs-CRP) levels were detected by circulating enzymatic method before treatment and 6-month after treatment. The WHO quality of life scale (WHOQOL-BREF) was used to assess the quality of life in two groups of patients. The improvements of angina and depressive symptoms were evaluated after 6-month treatment. The cardiovascular events were followed up in two groups. Re?sults HAMD scores significantly decreased after treatment in both groups (P < 0.05). HAMD scores at all different time points were significantly lower after treatment than those before treatment in treatment group. In control group, HAMD scores at different time points were also significantly lower after treatment than those before treatment, except time point of one week after treatment. The levels of Hcy and hs-CRP were decreased after treatment in two groups, and the levels were lower in treatment group than those of control group. Values of WHOQOL-BREF score, mental status, social relations and the sur?rounding environment scores were higher after treatment in both groups. Also they are higher in treatment group than that of control group. The angina and depressive symptoms were improved in treatment group. The incidences of angina, myocardial infarction, arrhythmia, heart failure, sudden cardiac death and other cardiovascular events were significantly lower in treat?ment group than those of control group (χ2=9.396, P<0.05). Conclusion Escitalopram oxalate combined with psychological therapy shows a significant beneficial effect and a better prognosis in the treatment of coronary heart disease and depression.