AIM:To investigate the effect of miRNA-181a inhibition on the proliferation, migration and cell cycle of the human multiple myeloma cell line RPMI 8226.METHODS:Real-time PCR was used to detect miRNA-181a expression in serum samples from multiple myeloma or healthy subjects .After transfection with miRNA-181a inhibitor, the cell viability was examined by CCK-8 assay and colony formation assay .The cell migration ability was analyzed by wound healing assay .The cell cycle was detected by flow cytometry .Moreover , the protein level of cyclin D 1 and the phosphoryla-tion of PI3K and Akt were determined by Western blot .RESULTS:The expression of miRNA-181a was significantly in-creased in the serum from multiple myeloma patients as compared with healthy group .Inhibition of miRNA-181a expression by transfection with miRNA-181a inhibitor remarkably decreased the cell viability , migratory ability, the population of G0/G1 phase and cyclin D1 protein expression in the RPMI8226 cells.However, the population of S phase and the phosphory-lation of PI3K and Akt were reduced .CONCLUSION:Down-regulation of miRNA-181a inhibits the viability and migra-tory ability in the RPMI8226 cells via inhibition of cell cycle and PI 3K/Akt signaling pathway .

0. 05). Conclusion: The HCAPl can be expressed in both normal peripheral white blood cells and the leukemic cells, and there is no difference in the protein levels between them.

Objective To evaluate the growth inhibition of human gastric carcinoma cell lines SGC 7901 in vitro and the expression of bcl-2, bcl 2l12 and bax with docosahexaenoic acid (DHA) and 5-fluorouracil (5-FU). Methods The effect of DHA and 5-FU was measured by trypan blue, and the interaction between two agents was judged by combination index (CI). Cells were observed by inverted microscope. Flow cytometry was used for analysis of apoptosis by PI staining and Annexin-V/PI. RT-PCR was used to analyze the levels of bcl-2, bcl 2l12 and bax mRNA. Results DHA significantly inhibited the growth of SGC 7901 cells in a dose- and time-dependent way ( P < 0. 05 ), the IC50 of 24 h and 48 h was 67. 81 μg/ml and 45.76 μg/ml, and a strong synergism was found in the combination of DHA and 5-FU (CI < 1 ,P <0. 01 ). Treated by DHA and 5-FU for 48 h, cells became sparse under inverted microscope. DHA or 5-FU was able to induce apoptosis and the effect became even more significant by the combination of DHA and 5-FU. Cells were holted in phase of G01/G1 and S. RT-PCR showed that DHA or 5-FU down-regulated the expression of bcl-2 and bcl 2l12 mRNA, while bax mRNA expression was not downregnlated. Conclusions DHA could inhibit the growth of gastric carcinoma cells, DHA and 5-FU had synergetic effect in the inhibition of the cells growth and blockage of the cell cycles possibly by down-regulating the expression of bcl-2 and bcl 2l12.

AIM:To detect the treatment of K562 leukemia cells with bortezomib altering the expression of genes fas,bcl-2,bcl2l12,bim,bax,caspase-9 and caspase-3.METHODS:MTT assay was used to detect the inhibition of proliferation.Apoptosis was detected by Annexin-V staining and mitochondrial transmembrane potential(??m).RT-PCR was used to analyze the mRNA expressions of fas,bcl-2,bcl2l12,bim,bax,caspase-3 and caspase-9.RESULTS:Bortezomib caused a time-and dose-dependent inhibition of cell proliferation and IC50 of 24 h and 48 h were 161.41 nmol/L and 96.33 nmol/L,respectively.At the concentration of 104 nmol/L,bortezomib induced apoptosis in a time-dependent manner,including increasing annexin-V positivity and decreasing the ??m.RT-PCR showed that bortezomib up-regulated the mRNA expression of fas,bcl2l12,caspase-9 and caspase-3,but mRNA expressions of bcl-2,bim and bax did not changed obviously.CONCLUSION:Bortezomib inhibits the proliferation of K562 and induces apoptosis,in which fas,bcl2l12,caspase-9 or caspase-3 gene is one of the main genes taking part in.

OBJECTIVE To study characteristics of changing T lymphocytes in epidemic hemorrhagic fever(EHF) patients during acute phase and find out the pathogenesis,in order to elevate the level of early diagnosis.METHODS The anticoagulant blood from 30 cases of EHF patients and 50 normal healthy blood donors was collected.T lymphocyte subsets were detected by flow cytometry.RESULTS Compared with those of normal persons,CD4+ T cell counts of EHF patients decreased,CD8+T cell and double CD4+CD8+ cell(double positive cells,DP cell) counts of EHF patients increased obviously,and 25 cases of EHF in recovery stage returned to normal.And in comparison with HIV,CMV and EBV patients,DP cell counts of EHF patients increased obviously.CONCLUSIONS T lymphocytes of EHF decrease obviously but could be resumed,detection of amounts of lymphocyte subsets and CD4+CD8+ cells can provide an early diagnosis method to EHF.

OBJECTIVETo explore the expression and clinical significance of Musashi2 (MSI2) gene in de novo acute myeloid leukemia (AML).

METHODSReal-time quantitative PCR (RQ-PCR) was used to measure the expression of MSI2 gene in 181 de novo AML patients. Correlation between the expression level and clinical features of such patients was explored.

RESULTSTranscript of the MSI2 gene was detected in 181 AML patients, with the median expression level being 2.341 (0.1124-58.8566). By contrast, CD34+ cells from 10 healthy controls had a much lower expression level (P=0.012), and the expression level of MSI2 in 24 patients with complete remission was significant lower than de novo patients (P=0.021). Based on the median expression level, such patients were divided into low expression group and high expression group. Patients from the high expression group had significantly higher rate of high white blood cell count (78% vs. 63%, P=0.034). Compared with MSI2-low group, FLT3-ITD mutation were much more common in MSI2-high group (28% vs. 7%, P=0.002). The expression level of MSI2 in aberrant karyotypes was much higher than that in favorable karyotypes (the median expression level was 2.7726 and 2.0733, P=0.035). Kaplan-Meier analysis showed that the overall survival in high expression group of MSI2 was lower than the low expression group, with the median survival time being 28 months and 12 months, respectively (P=0.045).

CONCLUSIONDe novo AML patients have a higher level of MSI2 gene expression. And the latter is much more common in those with high white blood cell count and aberrant karyotypes, and has a positive correlation with FLT3-ITD mutation. High expression of MSI2 gene may be a predictor for poorer prognosis among AML patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; RNA-Binding Proteins ; genetics ; metabolism ; Young Adult

OBJECTIVE: To explore the correlation of genetic mutations and clinical features of myelodysplastic syndromes (MDS) with scores of Revised International Prognostic Scoring System (IPSS-R). METHODS: Eighty-seven patients with de novo MDS were enrolled. Mutations of MDS-related genes and clinical features were used to determine the incidence and subtype of mutations. Clinical features and IPSS-R scores of the patients with high frequency mutations involving TET2, TP53, ASXL1, RUNX1 and SF3B1 genes were compared. RESULTS: Fifty-four patients (62.1%) harbored at least one point mutation. The incidences of various mutations were significantly different, with the incidence of MDS-EB-2 being 100% and MDS-SLD being only 38.9%. Compared with the wild types, patients harboring mutations had higher lactate dehydrogenase, higher β2 microglobulin, higher percentage of bone marrow blast cells and lower hemoglobin levels (P=0.027, <0.01, <0.01, 0.046, respectively). The IPSS-R scores of MDS patients with mutations were significantly higher than the wild types (P<0.01). The IPSS-R scores of the TP53 mutation groups were 7.82±1.83, which was significantly higher than the control group (3.77±1.66, P<0.01). No difference was found between the IPSS-R between patients carrying TET2, ASXL1, RUNX1, and SF3B1 mutations or the wild types (P>0.05). CONCLUSION Genetic mutations are commonly found in MDS. MDS patients with mutations have unique clinical laboratory characteristics. Although the prognostic value of most genes is controversial, TP53 is an definite indicator of poor prognosis.
DNA Mutational Analysis ; Humans ; Incidence ; Mutation ; Myelodysplastic Syndromes ; genetics ; Prognosis ; Tumor Suppressor Protein p53 ; genetics

Objective To investigate the genetic characteristics of patients with thalassemia gene in Ningbo City.Methods Totally 313 patients with thalassemia gene diagnosed during March 2015 to April 2016 in Ningbo First Hospital were included in the study.The results of routine blood test,hemoglobin electrophoresis and gene test,as well as the gender and origin distribution of patients with thalassemia gene were analyzed.Results Of the 313 patients who carried the thalassemia gene,there were 289 females and 24 males with a median age of 29 years,ranging from 1 d to 57 years after birth.And of the 313 patients,75 carried the α-thalassemia gene,230 β-thalassemia and 8 composite thalassemia.The average value of hemoglobin was around 100 g/L and the average value of erythrocyte mean corpuscular volume (MCV) was less than 80 ft.Abnormal hemoglobin was usually found in α-thalassemia.81.74％ (188/230) of β-thalassemia had abnormal hemoglobin electrophoresis.Most of the patients were women who were diagnosed of anemia in routine prenatal examination.The number of patients,who came from Ningbo City or one of whose grandparents came from Ningbo City,was closed to 50％.Among 20 α-thalassemia patients coming from Ningbo City,genotype of--sea was the commonest genotype,accounted for 70.00％ (14/20).Among 82 β3-thalassemia patients coming from Ningbo,genotype of IVS-Ⅱ-654 was commonest genotype,accounted for 54.88％ (45/82).Genotypes of 2 composite thalassemia coming from Ningbo City were αcs compound IVS-Ⅱ-654 and-α3.7 compound CD41-42.Conclusions In Ningbo City,the incidence of thalassemia in women in Ningbo is higher than that in men.The morbidity of β-thalassemia genotype is apparently higher than that of α-thalassemia,and genotype of IVS-Ⅱ-654 in β3-thalassemia patients is the commonest genotype.

OBJECTIVE: To explore the clinical features and genomic abnorm ality of a fetus enlarged multicystic dysplastic kidneys with oligohydramnios caused by NPHP3 gene mutation. METHODS: The fetuse was found to have multicystic dysplastic kidneys with oligohydramnios upon ultrasonography during the second trimester. Following induced abortion, fetal tissue was collected for the extraction of DNA, chromosomal microarray analysis (CMA) and whole exome sequencing (WES). Sanger sequencing was used to verify the suspected variants in the family. RESULTS: Antenatal ultrasound examination at 19 weeks showed "polycystic" kidneys with Oligohydramnios. Delivery was by induced labour because of the critically low amniotic fluid volume. Testing of CMA was normal. WES showed a compound heterozygous mutation of c.1817G>A, p.W606X; c.432dupA, p.E145Rfs*18 mutations are novel mutations in this study. CONCLUSION The research may further expand the NPHP3 gene mutation spectrum. Enlarged multicystic dysplastic kidneys with oligohydramnios caused by NPHP3 gene mutation at least include one or two splice site mutation, frameshift mutation or nonsense mutation foetal poor prognosis.
Amniotic Fluid ; Female ; Humans ; Kidney Diseases, Cystic ; Multicystic Dysplastic Kidney/genetics* ; Mutation ; Oligohydramnios/genetics* ; Polycystic Kidney Diseases ; Pregnancy ; Ultrasonography, Prenatal

Objective:This study is aimed to investigate the value of absolute lymphocyte count (ALC) in predicting the clinical prognosis of patients with myelodyplastic syndrome(MDS).Methods:245 patients with MDS who diagnosed in our hospital from 2009 to 2019 were analyzed retrospectively, re-diagnosed according to WHO 2016 standard, and 208 patients with intact IPSS-R were risk-stratified, all of the patients′ peripheral blood ALC were collected and analyzed, through the time dependent receiver operating characteristic curve (ROC) analysis in Survival ROC package of R language, the optimal threshold value of ALC was 1.0×10 9/L. The patients of MDS were divided into normal ALC group (ALC ≥1.0×10 9/L) and low ALC group (ALC<1.0×10 9/L). Pearson χ 2 test and Mann-Whitney U test was used to analyze the differences in general data between the two groups. The overall survival (OS) curve and leukemia-free survival (LFS) were plotted by Kaplan-Meier method and compared by Long-rank test. Factors influencing the prognosis of MDS were analyzed by Cox Regression Model. Results:There were 97 cases in low ALC group and 148 cases in normal ALC group. The low ALC group had lower OS (15 months vs 60 months, P<0.000 1) and higher IPSS-R score (5.0 vs 3.75, P = 0.001). Multivariate analysis showed that ALC (<1.0×10 9/L) (HR:0.374,95% CI:0.153-0.917, P = 0.032) was independent risk factor of OS in IPSS-R-intermediate-risk MDS patients. Conclusion:This study shows that ALC in peripheral blood is an independent risk factor in IPSS-R-intermediate-risk MDS patients, which provides clinical evidence for the influence of body immunity on the development of MDS.