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 .

Objective To determine the expression of CD24 in colorectal carcinoma, and to explore the relationship between CD24 and the clinicopathological features of colorectal carcinoma.Methods The expression of CD24 in 62 cases of colorectal carcinoma, 47 cases of adenomas, 15 cases of colorectal polyps and 30 cases of the adjacent non-cancerous tissues were observed by immunohistochemical assay.The relationship between CD24 and the clinicopathological features was analyzed.Results The positive rates of CD24 in colorectal carcinoma 72.6％ and adenomas 63.8％ were significantly higher than those in colorectal hyperplastic polyps 13.3％ (x2 =17.83, P =0.00;x2 =11.61, P =0.00) and adjacent non-cancerous tissues 6.7％ (x2 =35.15, P =0.00;x2 =24.64, P =0.00).The expression of CD24 in colorectal carcinoma had a significant correlation with the tumor diameter (x2 =5.48, P =0.02), tumor differentiation (x2 =8.86, P =0.00), Duke staging (x2 =11.47, P =0.00) and lymph node metastasis (x2 =8.92, P =0.00).Conclusion The expression of CD24 is high in colorectal carcinoma, having a significant correlation with the size of tumor, degree of differentiation, Duke stage and lymph node metastasis.

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.

AIM:To study the effect of vitamin K3(VK3) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS:Cell viability was estimated by MTT assay.AO/EB staining was performed to detect apoptotic cells.Apoptosis and the changes of cell cycle were detected by flow cytometry.NAC was used to observe the effect of growth inhibition by VK3.RT-PCR was used to confirm the changes in gene expression.Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA.RESULTS:PC-3M cells growth was significantly inhibited by VK3(≥60 ?mol/L,P

Objective To establish a optimal method and threshold of 3-deoxy-3-fluorothymidine (FLT) PET-CT in delineating the biological target length of gross tumor in esophageal carcinoma, and to compare FLT PET-CT with other imaging modalities including esophagoseopy, esophagography, CT and flu-orodeoxyglucose (FDG) PET-CT. Methods Twenty-four patients with esophageal squamous cell carcinoma treated with radical surgery were enrolled. Before surgery, all the patients underwent FLT PET-CT, esepha-goscopy and esophagography. Twenty-two patients also received FDG PET-CT scan. Gross tumor volumes (GTV) were delineated using seven different threshold of FLT PET-CT: visual interpretation, standardized uptake value (SUV) 1.3, SUV 1.4, SUV 1.5, 20% of maximum standard uptake value (SUV_(max)), 25% SUV_(max), and 30% SUV_(max). Three different thresholds of FDG PET-CT were used, including visual interpre-tation, SUV 2.5, and 40% SUV_(max). The length of tumors on FLT PET-CT scan were measured and recorded as L_(FLTvis), L_(FLT1.3), L_(FLT1.4), L_(FLT1.5), L_(FLT20%), L_(FLT25%), and L_(FLT30%), respectively. The length of tumors on FDG PET-CT scan were recorded as L_(FDGvis), L_(FDG2.5), and L_(FDG40%), respectively. The length of tumors on CT, esophagography and esophagoscopy were recorded as L_(CT), L_(X-ray) and L_(Scopy). All of these results were com-pared with the length of gross tumor in the reseeted specimen measured by pathological examination (L_(Path)), Results The L_(Path) was (4.90±2.14) cm. The Length of tumors delineated by different methods, being from short to long, were L_(FDG40%), L_(Scopy), L_(X-ray),L_(FLT1.5),L_(CT),L_(FLT30%),L_(FLTvis),L_(FLT1.4),L_(FLT25%), L_(FDG2.5),L_(FDGvis),L_(FLT1.3),L_(FLT20%). The mean values were (3.85±1.52), (4.46±2.23), (4.63± 2.37), (4.64±2.38),(4.69± 1.85),(4.75±2.19) ,(4.85±2.33),(4.87±2.35),(5.05±2.20), (5.08± 2.19) ,(5.10±2.22), (5.21±2.40) and (5.53±2.17) cm,respectively. The correlation coefficients were 0.91,0.93,0.88, 0.95, 0.90, 0.81,0.96, 0.96, 0.80, 0.99, 0.99, 0.95 and 0. 79 , respective-ly. All the P values were 0. 000. L_(FLT1.4) of FLT PET-CT and L_(FDG2.5) of FDG PET-CT were found more ap-proximate to L_(Path). There was no significant difference between L_(FLT1.4) and L_(FDG2.5) (1= 1.23, P = 0.232), and the correlation coefficient was 0.96 (P = 0. 000). Conclusions Thresholds of SUV 1.4 on FLT PET-CT and SUV 2.5 on FDG PET-CT could optimally estimate the tumor length measured by pathological examina-tion, and could be objective and simple methods for semiquantitative analysis.

Objective:To detect precancerous lesions of gastric cancer and biopsy tissue vascular endothelial growth factor (VEGF)and mutant p53 gene(mtp53)expression,to explore the development of clinical significance of VEGF and mutant p53 gene in gastric cancer.Methods:19 cases by endoscopic biopsies of normal gastric tissues,22 cases of intestinal metaplasia,47 cases of gastro-intestinal mucosal dysplasia, 54 cases of gastric cancer samples by immunohistochemical staining to detect the expression levels of VEGF and mtp53′s.Results: The expression levels of VEGF, mtp53 in normal gastric mucosa, intestinal metaplasia, dysplasia, and gradually increased gastric cancer was the law.mtp53 of VEGF expression in gastric carcinoma and compared with normal gastric tissue,intestinal metaplasia was significantly higher(P<0.05),but with atypical hyperplasia was no significant difference(P>0.05). Conclusion: The abnormal expression of VEGF and mutant p53 may be related to the degree of deterioration of the stomach tissue lesions related.

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 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: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.