Objective To analyze the treatment efficacy and safety of a modified GMALL protocol for adult acute lymphoblastic leukemia (ALL). Methods Data of 37 patients with newly diagnosed adult ALL treated with a modified GMALL protocol from January 2005 to December 2009 were retrospectively analyzed,and compared with that of 44 patients treated with an in-house conventional protocol at the same period.Results The complete remission (CR) rate was 89.2 %(33/37) treated with modified GMALL protocol. The cumulative overall survival (OS) rates at 1 year, 2 years, 3 years and 4 years were 77.5 %, 48.0 %, 40.0 %and 40.0 %, respectively. The main adverse events were grade 3 or grade 4 hematological toxicities and infections which were easily managed, and the treatment-related mortality rate was low. The OS of modified GMALL protocol was superior to that of the conventional protocol. Conclusion The modified GMALL protocol has a satisfying effect and the adverse events can be tolerated for adult ALL, so its clinical application can be encouraged.

Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10%FBS, during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation, including three-drug combination group and four-drug combination group. The protocol of mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination induced differentiation, four-drug combination was successfully achieved in over 90% of the cell inducing, which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.

Objective To assess the capability of monochromatic energy images of Gemstone spectral imaging(GSI) in reducing artifacts caused by metallic implants. Methods Twelve subjects with metallic implants underwent GSI (Discovery CT750 HD, GE Healthcare, Milwaukee ). The metallic orthopedic implants included 3 patients of dentures, 2 patients of cervical spinal vertebraplasty, one clavicle fracture fixation, one lumbar spinal vertehraplasty, 3 patients of artificial femoral head, one iliac fracture fixation and one tibial fracture fixation. GSI was performed by using a single source ultra-fast dual energy X-ray switching (80 kVp and 140 kVp). Following GSI scanning, thin slice images were reconstructed into 1.25 mm slice thickness. The monochromatic energy images were set to the same window width and level (window width 1500 HU,window level 500 HU). The artifact indexes (AI) at different kiloelectronvolts (keV) images were measured and compared. 3D reconstruction was performed using images with minimal AI. Result The artifacts index on monochromatic energy images varied with the change of keV. Of the images from 12 subjects, the maximal AI ranged between 145-225 at 40 keV, and minimal AI ranged between 15-90 at the 95-140 keV. The artifacts are clearly visible on polychromatic energy images and the artifacts are reduced markedly on the monochromatic energy images with minimal AI. Conclusion The artifacts caused by metallic implants can be reduced significantly by GSI with high keV monochromatic energy images.

Objective:To investigate the correlations of Lauren classification and world health organization (WHO) classification of gastric cancer (GC) with microvascular density (MVD), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2), and p53. Methods:The clinical data of 89 patients with GC were collected. The collected specimens were categorized on the basis of Lauren classification and WHO classification. CD34/periodic acid-Schiff (PAS) double staining was performed to validate MVD. Immunohistochemistry was conducted to investigate the expression levels of MMP-2, MMP-9, VEGF, VEGFR1, VEGFR2, and p53. Results:MVD was not correlated with Lauren classification or WHO classification (P>0.05). Lauren typing was associated with the expression levels of MMP-9, VEGFR1, and p53 (P<0.05). WHO classification was not related to any of the factors (P>0.05). Cox proportional hazards model revealed that Lauren classification and WHO classification were the prognostic factors of overall survival (P<0.05). Conclusion:This research on tumor related factors, angiogenesis, and different classifications of GC may provide new methods to treat this disease.

Objective To explore whether hypoxia could promote epithelial-mesenchymal transition (EMT) in various differentiated colorectal cancer cells, and analyse the effect of hypoxia on invasion and migration of colorectal cancer cells. Methods HCT116 (poorly differentiated) and HT-29 (highly differentiated) colorectal adenocarcinoma cells were selected respectively. The morphological changes of two cell lines were observed after 0,10,25,50,100 and 150 mg/L cobalt chloride (CoCl2) treatment for 48 h. The expression of hypoxia-inducible factor-1α(HIF-1α) protein was analysed after 0, 10,25,50,100 and 150 mg/L CoCl2 treatment for 48 h. An optimal concentration of CoCl2 was then selected. Methylthiazolyl tetrazolium (MTT) assay was used to detect the proliferation of two kinds of colorectal cancer cells induced by CoCl 2 at different time points (0, 24, 48, 72 and 96 h), and to select an optimal time. Under the optimal concentration and time conditions, the HCT116 and HT-29 cells were processed by hypoxia (hypoxia group) and normoxia (normoxic group). Transwell invasion assay and Wound healing assay were used to detect cell invasion and migration in two groups. Western blot assay and RT-PCR were used to detect protein and mRNA expression levels of HIF-1α, E-cadherin and Vimentin in two groups. Results Two kinds of cells showed obvious morphological changes after 50 mg/L CoCl2 treatment for 48 h. HIF-1αprotein level first increased and then decreased in two groups of cells with the increased concentration of CoCl 2, and 50 mg/L CoCl2 was the optimal concentration (P<0.05). The cell proliferation showed a tendency to decrease after the increase in both kinds of cells with or without hypoxia for 0-96 h (P<0.05), and 48 h was the optimal time. The transmembrane number and cell migration rate were significantly more in hypoxia group than those of normoxic group (P<0.05). The protein and mRNA levels of HIF-1α and Vimentin were significantly higher in hypoxia group than those of normoxic group in HCT116 and HT-29 cell lines (P<0.05). E-cadherin protein and mRNA levels were significantly lower in hypoxia group than those of normoxic group (P<0.05). Conclusion Hypoxia can promote EMT in different differentiated colorectal cancer cells, and can enhance invasion and migration of two kinds of colorectal cancer cells.

Objective To set up a real-time quantitative PCR approach for detection and quantification for bcr-abl transcripts in CML patients,and detect minimal residual disease (MRD) in CML by real-time quantitative PCR (RQ-PCR)and evaluate the significance of MRD detection.Methods The ber-abl.fusion gene expression in 80 patients with CML was analyzed by RQ-PCR. The patients were divided into three groups according to the different treatment, allogeneic hematopoietic stem cell transplantation group,imatinib group and hydroxyurea group. The change of bcr-abl fusion gene was monitored in CML patients before and after treatment.Results The average of RQ-PCR detection on newly diagnosed patients with CML in chronic phase was 6847.67 copies / 104 cells,the accelerated phase was 306 176.08 copies / 104 cells,and the average results were 944.33, 2.37, 0.29, 0 copies / 104 cells after allogeneic hematopoietic stem cell transplantation one month,6 months,12 months or 24 months respectively.The average of RQ-PCR detection after use imatinib mesylate 3 months was 3720.23 copies / 104 cells and not be detected after one year. The average was 7290.11 and 3143.24 copies / 104 cells after hydroxyurea treatment 0 and 9 months respectively.The difference in first two groups was not significant (t=1.74,P=0.17), but the difference between the third group and the first two groups was significant (t=3.74,P=0.01.t=2.97,P=0.02). The upregulation of bcr-abl transcript levels could be detected when disease progression. The transcripts level in accelerated phase was significantly higher than that in chronic phase. Conclusion RQ-PCR can be used to detect the MRD,monitor the treatment outcome,predict disease recurrence and give early intervention.

Objective:To discuss the influence of ALDH1+and CD133+phenotypic breast cancer stem-like cells in TA2 triple negative breast cancer on promoting epithelial-mesenchymal transition (EMT) occurrence in TA2 mice with triple-negative breast cancer and on their biological behavior. Methods:Flow cytometry was performed to analyze the markers ALDH1 and CD133 in TA2 mice triple nega-tive breast cancer and breast cancer stem-like cells with ALDH1+, ALDH1?, CD133+, and CD133?phenotypes, which were sorted out. Then, the TA2 mice were inoculated with sorted tumor cells according to cell type. The mice were divided into ALDH1+, ALDH1?, CD133+, and CD133-groups. The tumor-growing conditions were observed. A tumor tissue was sliced for the immunohistochemical testing of ALDH1?, CD133?, and EMT-related Twist1, E-cadherin, and VE-cadherin proteins. The expression difference of breast cancer stem cell surface markers ALDH1 and CD133 in triple-negative breast cancer and EMT-related proteins Twist1, E-cadherin, and VE-cad-herin was analyzed. Results:The expression rates of breast cancer stem cell markers ALDH1 and CD133 in TA2 mice triple negative breast cancer were 31.2%and 6.5%, respectively. The tumor growth ability of TA2 mice from ALDH1+group was obviously stronger than that from ALDH1?group. The CD133+group was evidently stronger than CD133?group. The immunohistochemical results showed that ALDH1, Twist1, and VE-cadherin expression levels in the ALDH1+group were evidently higher than that in the ALDH1?group (all P<0.05). E-cadherin expression decreased (P<0.05). CD133?, Twist1, and VE-cadherin expression levels in CD133+group were higher than that in CD133?group (all P<0.05). Conclusion:In TA2 mice triple negative breast cancer, ALDH1+and CD133+phenotypic breast cancer stem-like cells may influence the expression of EMT-related proteins, and promote the formation of triple-negative breast cancer.

ObjectiveTo explore the genetic abnormalities of multiple myeloma (MM)patients by fluorescence in situ hybridization (FISH).MethodsWith the application of FISH,sequence specific DNA probes (IGH,DI3S319/p53 and 1q21/RB1) were applied to detect 14q32 rearrangement,del(13q14),del (17p13)and gain of lq21.Forty-four MM patients were enrolled in this study.ResultsThirty-two cases (72.7 ％) detected by FISH had genetic abnormality in 44 cases,lq21 amplification was observed in 11 cases (25.0 ％),while RB1 deletion in 17 cases (38.6 ％),D13S319 deletion in 16 cases (36.4 ％),p53 deletion in 6 cases(13.6 ％)and 14q32 translocation in 19 cases(43.2 ％).The patients with one abnormality was detected in 10 cases(22.7 ％),two abnormalities in 11 cases(25.0 ％),three abnormalities in 8 cases (18.2 ％),4 abnormalities in 3 cases(6.8 ％).28 were found to undergo split-phase by conventional cytogenetic in 44 patients.The patients with genetic abnormalities detected by conventional G-banding was 2 cases (7.14 ％),the difference with that in FISH was significant (P ＜0.05).Genetic abnormalities compared with clinical parameters showed that β2-MG in IGH gene abnormal patients were significantly higher than those without such abnormalities (P ＜0.05).Patients with bone marrow plasma cells of lq21 amplification were higher than those with normal karotypes(P ＜0.05),CRE was significantly higher among lq21 amplification and p53 deletion patients (P ＜0.05),CRP was significantly higher among p53 deletion patients (P ＜0.05).No significant difference was oberserved in relationship of the chromosome aberration and age,the chromosome aberration and stage.ConclusionThe most common genetic abnormalities in MM is IGH rearrangement and absence of RB1 and D13S319,followed by lq21 amplification,the least is p53 deletion.FISH is a rapid and sensitive technique to refine chromosome aberrations in MM.The specific detection for genomic features of MM is proved to be correlative with its clinicopathologic characteristics and the prognosis.

Objective:To study the predictive value of the amplitude-integrated electroencephalography (aEEG) within 6 hours and 3 days after birth and magnetic resonance imaging(MRI) on the adverse neurobehavioral development of asphyxiated preterm infants at the correction age of 6 months.Methods:From December 2017 to June 2019, 50 asphyxiated preterm infants who were delivered at the obstetrical department transferred to the division of neonatology in the Third Affiliated Hospital of Anhui Medical University were monitored by aEEG within 6 hours after birth, then once a day for at least 4 h. MRI was administered at 40 weeks of corrected age, neuromotor developmental function of the infants was assessed by the Geisel developmental diagnostic scale at 6 months of corrected age, then the infants were divided into good prognosis group and poor prognosis group according to the assessment results. SPSS 19.0 software was used for statistical analysis.The software of SPSS 19.0 was used to analyze the data.Independent sample t-test and χ 2 test were used to analyze the difference between the two groups.The relationship between aEEG grading and MRI, and their predictive value for adverse neurobehavioral development were analyzed at 6 months of corrected age. Results:The degree of white matter damage( H=24.896) and intracranical hemorrhage( H=29.245) of premature infants with different aEEG clinical grades were different (both P<0.01) on MRI. The sensitivity of aEEG within 6 hours and 3 days after birth on predicting poor prognosis was 96.2% and 97.8%, the specificity was 56.2% and 62.5%, the negative predictive value was 98.2% and 99.0%, the positive predictive value was 37.8% and 52.3%, the correct index was 52.4% and 60.3%, respectively. The aEEG was combined with MRI, the sensitivity (90.0%, 97.0%), specificity (89.0%, 99.0%), negative predictive value (99.2%, 99.5%), positive predictive value (80.6%, 88.5%), and correct index (79%, 96%) were all improved. Conclusion:The combination of aEEG grading and MRI can improve the prognostic value on neurodevelopmental prognosis, and provide a better evaluation basis for clinical follow-up and intervention of asphyxiated premature infants with brain injury.

Objective:To examine the expression of IQGAP1 in hepatocellular carcinoma and its effect on vasculogenic mimicry(VM). Methods:Immunohistochemical staining was performed to investigate the expression of IQGAP1.CD31/PAS double staining was per-formed to detect VM and analyze the correlation of IQGAP1 and VM.HepG2 cells were transfected with an IQGAP1 overexpression plasmid to induce exogenous expression of IQGAP1,and an IQGAP1 knockdown plasmid was transfected into SMMC7721 cells to re-duce IQGAP1 levels.The expression of cancer stem cell markers CD133,CD44,Sox2,and ALDH1 was analyzed by Western blot and compared with that in the control.Cellular functional experiments were used to determine the role of IQGAP1 in promoting cancer cells' ability of invasiveness and migration, proliferation, and VM formation. Results: Immunohistochemical analysis revealed that IQGAP1 was mainly located in the cell membrane and/or cytoplasm,and the staining intensity was correlated with tumor grade,me-tastasis,and VM(P<0.05).Cells transfected with the overexpression plasmid showed enhanced CD133,CD44,Sox2,and ALDH1 levels due to the increase in IQGAP1 and exhibited increased invasion ability,proliferation,and VM formation.In the SMMC7721 cells trans-fected with the knockdown plasmid,CD133,CD44,Sox2,and ALDH1 levels were decreased and motility was inhibited.Conclusions:IQGAP1 supports malignant behavior in hepatocellular carcinoma and may promote VM by increasing stemness.