Objective To study the quantification of MDR1 and WT1 gene expression in patients with de novo acute myeloid leukemia(AML)and to explore the role of these two genes in clinical drug resistance and their correlation with risk stratification. Methods A real time quantitative reverse transcriptase polymerase chain reaction method was established for detecting MDR1 and WT1 gene expression levels in 63 de novo AML patients.Resuits The expression of WT1 and MDR1 was significantly higher in de novo AML patients than in normal controls (P＜0.001).WT1 levels were significantly correlated with corresponding levels of MDR1 gene in de novo AML patients(P=0.004).Expression levels of WT1 and MDR1 gene were not associated with FAB subtype and risk stratication(P＞0.05).AML patients with FLT3-ITD mutations had a significantly higher WT1 expression level as compared to with those without(P＜0.05),on the contrary MDR1 expression was not associated with FLT3-ITD mutations(P＞0.05).Patients with co-expression of high levels of WT1 and MDR1 had a significantly lower complete remission rate after induction therapy than those with low levels(P＜0.05).Conclusion There is a positive correlation between MDR1 gene expression and WT1 gene expression in AML.Quantification of the two gene expression together is more effective for judgement of prognosis in AML.

Antineoplastic Agents, Hormonal ; therapeutic use ; Bone Neoplasms ; secondary ; Breast Neoplasms ; chemistry ; drug therapy ; pathology ; Disease Progression ; Female ; Humans ; Menopause ; Neoplasm Recurrence, Local ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis ; Tamoxifen ; therapeutic use

Objective To evaluate the clinical application of the fistulography of multi-slice spiral CT(MSCT)in the diagnosis of anal fistula.Methods A total of 28 patients who were verified operatively of fistula in ano underwent preoperative fistulography of MSCT.The multi-planar reformation(MPR)and surface shadow display(SSD)images were generated and analyzed retrorespectively.Results There were 8 cases with perianal abscess and 20 cases with anal fistula in 28 patients.Complex fistula was diagnosed in 16 cases,and simple inter-sphincteric fistula was found in 4 cases;the inner orificiums of anal fistulas were revealed accurately with MPR images in 18 cases.Conclusion The fistulography of MSCT is valuable for preoperative assessment of anal fistula,in particular for investigation of the inner orificiums of anal fistulas with MPR.

Objective To investigate the sensitivity of BIOMED-2 primer system in adult acute lymphoblastic leukemia (AIJ,) patients Ig gene rearrangement, and to analyze their frequency, corearrangement pattern, utilization of V, D and J genes and composition of junctional regions. Methods Amplification of rearranged IgH (complete and incomplete), IgK, IgK-Kde and IgL was performed in standard PCR in 29 adult ALL patients. Monoclonal PCR products were subjected directly to DNA sequencing. Sequences were identified by comparison with all known human Ig germline sequences to analyze the recombination patterns, somatic mutations and germline gene segments usage. Results IgH, incomplete IgH, IgK, igK-Kde and Igl, rearrangements were found with positive rate of 70.8%, 12.5% , 29.2% , 25.0% and 0 of B-ALL patients, respectively. All B-ALL patients displayed at least one pattern of Ig gene rearrangements. In TALL, one of five patients was found with incomplete IgH rearrangement, two patients were found with IgK rearrangements and two patients were PCR-negative. The sequence analysis showed that the most frequently used V, D, J segments in adult B-ALL patients were from VH3/VH4 families, DH3 family and JH6 family, respectively. Four of five IgK rearrangement used VκI family. 23.5% B-ALL IgH contained scattered replacement mutations with replacement to silent substitution ratio < 1 in complementarity determining regions. Conclusion BIOMED-2 multiplex PCR analysis strategy is a reliable and useful technique in the adult BALL patients.

Objective To observe the effects of Chinese herbal compounds (alkaloid of Coptidis Rhizoma 40%, total saponin of panax notoginseng 35%, and polysaccharide of Ophiopogonis Radix 25%) on levels of blood glucose, fructosamine, and insulin of diabetic rats, and explore mechanism of action.Methods Ten rats were randomly chosen as blank control group, and other rats were used to establish diabetic rat models by STZ intraperitoneal injection. Successfully molding rats were randomly divided into Chinese herbal compounds high, medium, low dose groups, metformin group, and model group, 10 rats in each group. Each rat received gavage for continuous 4 weeks. Then rat blood was collected by cutting head and taking caudal vein. FBG was detected by glucose oxidase method;insulin level was detected by radio-immunity double antibody method;fructosamine content was detected by fructosamine method.Results Compared with the blank control group, model group the levels of FBG, froctosamine content increased, while fasting insulin level were lower after 4 weeks'medication. Compared with the model group, the levels of FBG, froctosamine content in Chinese herbal compounds high and medium dose groups and metformin group were lower, while fasting insulin level increased after 4 weeks'medication (P<0.05).Conclusion TCM herbal compounds can increased the basic insulin level by decreasing the contents of FBG and fructosamine.

OBJECTIVE: To evaluate the diagnostic value of multi-slice spiral CT (MSCT) for plasma cell mastitis. METHODS: Radiographs of MSCT for forty-six patients with plasma cell mastitis diagnosed by pathological examination were reviewed. RESULTS: The findings of MSCT of plasma cell mastitis could be divided into four types, including the inflammation type, the abscess type, the sinus and fistula type, and the mixed type, and each type had its radiographic characteristics. CONCLUSION: MSCT is helpful for diagnosing plasma cell mastitis and should be used as an examination of first choice for the patients.

BackgroundMonocyte chemotactic protein-1 (MCP-1)plays an important role in the tumor,inflammation,diabetic retinopathy and other neovascular disease,but the expression and the role of MCP-1 in the oxygen induced retinopathy(OIR) model have rarely been reported. Objective This study was to investigate the expression of MCP-1 in the retina development of newborn mouse and in mouse models with OIR.Methods C57BL/6J newborn mice were divided into two groups and 60 mice in each group.Mice in OIR group were exposed to 75％ oxygen for 5 days and then to room air.All mice in normal control group exposed to room air only.Ten mice in each group were randomly chosen and sacrificed at postnatal 5,7,12,14,17,21 days.The expression of MCP-1 in mouse retina was detected with the method of immunohistoehemistry and reverse transcription polymerase chain reaction(RT-PCR).Results MCP-1 positive cells were seen in normal mouse retina.Up-regulation of MCP-1 positive cells was detected both in 12 days in normal control group and in 14 days in OIR group.MCP-1 mRNA was detected in mouse retina at 5 days,and a transient up-regulation of MCP-1 mRNA was observed in 12 days in normal control group.MCP-1 mRNA in OIR group significantly increased in 14 days in comparison with the normal control group( P =0.028,P =0.001 ). Conclusions Expression of MCP-1 is detectable in whole retinal development procession of mice.A transient up-regulation of MCP-1 expression is detected in the critical period of retinal vascular development in mice models with OIR,which is closely related to the retinal vascular development and progression of retinal new vessels.

Polyamine biosynthesis is controlled primarily by omithine Decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC). Antisense ODC and AdoMetDC sequences were cloned into an adenoviral vector (Ad-ODC-AdoMetDCas). To study the inhibitory effects of Ad-ODC-AdoMetDCas on polyamine biosynthesis and esophageal cancer cell apoptosis, adenovirus-mediated gene tmnsduction efficiency was assessed with counting GFP-positive cells using MTT. The malignant phenotype of Eca109 cells was assessed by growth curve. Western blot and HPLC were used to detect ODC and AdoMetDC expression and polyamine content in Ecal09 cells. TUNEL was used to analyze cell apoptosis. The change of morphology of apoptotic cells was observed by electron microscope. It was demonstrated approximate 70% of Eca 109 cells were infected with Ad-ODC-AdoMetDCas when MOI reached 50. The expression of ODC was inhibited in the infected tumor cells. Ad-ODC-AdoMetDCas could inhibit Ecal09 cell growth and invasive ability. TUNEL proved that Ad-ODC-AdoMetDCas can lead to cell apoptosis. Characterized morphology was observed by electronmicroscope (ehromatincondensation,nuclear disintegration,formation of apoptoticbodies).It was suggested Ad-ODC-AdoMetDCas has significant inhibitory effects on esophageal cancer cell proliferation, leads to cell apoptosis and bears therapeutic potential for the treatment of esophageal cancer.

Objective To set up a multiplex real time quantitative PCR method to detect the expression of WT1 and MDR1 gene simultaneously in acute leukemia patient. Methods Total RNA was extracted from k562 cell line and was reverse transcribed to cDNA by the outer primers of WT1 and MDR1 respectively. The cDNA of WT1 and MDR1 were purified and digested by Bam H Ⅰ and Bgl Ⅱ , and then the two fragments were ligated to form the recombinant fragment WT1 + MDR1. The outer forward primer of WT1 and outer reverse primer of MDR1 were used to amplify the recombinant fragment WT1 + MDR1. The PCR product was purified and cloned into pMD18-T vector, and then transferred into E. coli DH-5α. A new kind of WT1-MDRl-contained standard plasmid was obtained from the positive colony. The recombinant plasmid was verified by digestion with restriction enzyme and PCR amplification. A multiplex real time quantitative PCR method was set up with FAM-labeled MDR1 probe and VIC-labeled WT1 probe in one reaction tube. The WT1 and MDR1 gene expression was detected in forty-seven AL patients and thirty-two controls by this method. Seven patients were followed-up to elucidate the relationship between the gene expression levels and clinical prognosis. Results The recombinant plasmid was confirmed by EcoR1 digestion and PCR amplification. The multiplex real time quantitative PCR technique could reach the sensitivity of WT1 and MDR1 gene up to 102 copy/μl. The standard curve slopes were 0. 999 and 0. 998. The WT1 [ 37 000( 163-6 370 000 )copies/μg RNA ] and MDR1 [ 76 200( 179-18 000 000 )copies/μg RNA ]expression levels of AL patients were significantly higher as compared to the controls [ 258( 0-643 ) copies/μg RNA and 333( 0-779 )copies/μg RNA ]( Z= 6. 755,6. 736, P ＜ 0. 01 ). Following up seven patients with similar regimen of chemotherapy, the WT1 and MDR1 expression correlated to the clinical course. Three AL patients with WT1 and MDR1 expression levels (2 170 and 86 900, 1 130 and 5 860, 1 170 and 586 copies/μg RNA )significantly decreased after chemotherapy and kept in the low range ( 370 and 560,138 and 980, 150 and 690 copies/μg RNA ), and had a favorable outcome. Three AL patients with WT1 and MDR1 expression levels ( 1 600 and 11 800, 24 800 and 968, 48 200 and 1 100 000 copies/μg RNA )decreased after initial chemotherapy, but increased significantly afterwards (20 314 and 25 660,184 364 and 31 530, 15 680 and 878 000 copies/μg RNA ),and suffered clinical relapse. One patient with high WT1 and MDR1 expression levels ( from 81 600 and 1 200 000 copies/μg RNA to 124 100 and 7 632 400 copies/μg RNA )showed the persistence of disease. Conclusions A multiplex real time quantitative PCR method to detect WT1 and MDR1 gene simultaneously is constructed successfully. The expression of WT1 and MDR1 may provide useful information for AL patients prognosis.

Objective To investigate BAALC(brain and acute leukemia cytoplasmic)gene expression in patients with de novo acute myeloid leukemia(AML)and its clinical significance. Methods BAALC expression was determined by real-time quantitative polymerase chain reaction(RQ-PCR) in 63 de novo AML patients.The association between BAALC expression and therapeutic effect was analyzed.Results The correlation coefficiencies were over 0.99 for standard curves of RQ-PCR method. BAALC expression was detected in 49(78%)AML patients.The peripheral WBC counts,hemoglobin, platelet counts and the bone mahow blast cell percentage at onset in 31 AML patients with high BAALC expression were(26.3?18.1)?10~9/L,(78.3?21.8)g/L,(76.9?64.5)?10~9/L and(61.2?22.3)% and those of 32 AML patients with low BAALC expression were(30.2?21.7)?10~9/L,(81.6?30.9)g/L, (73.9?57.2)?10~9/L,(54.3?16.3)%,respectively.No statistic differences were found between these two groups.The AML patients with normal chromosome karyotypes are more likely to have a high BAALC expression(68%)compared with those with abnormal chromosome karyotypes(23%,?~2=12.093,P= 0.001).AML patients with normal cytogenetics and high BAALC expression shows significant lower CR rate (65%)compared with those with low BAALC expression(84%,?~2=6.573,P=0.013). Conclusion High BAALC expression may define an important risk factor in AML with normal cytogenetics and predicts an adverse prognosis.