Objective To investigate the effect of Treg cells and Th17 cells on the pathogenesis of SLE in patients with SLE,by detecting levels of serum IL-10 and IL-17 in patients with systemic lupus erythematosus (SLE).Methods Selected 54 patients with SLE diagnosed in Suzhou Municipal Hospital Affiliated to Nanjing Medical University from July 2013 to July 2015 as subjects,and 19 healthy persons were selected as control group.Using double antibody sandwich method (ELISA) to detect the levels of IL-10 and IL-17 in two groups.Using indirect immunofluorescence and western blot method to detect ANA,dsDNA,RNP,Sm,SSA and SSB in patients with SLE.Then compared and correlation analysed the level of IL-10 and IL-17,the levels of IL-10 and IL-17 in ANA,RNP,Sm,SSA and dsDNA groups were analyzed simultaneously.Results The level of IL-10 and IL-17 in SLE were 63.7±89.0 pg/ml and 87.7±123.0 pg/ml,and control group were 20.8±8.9 pg/ml and 18.5 ± 111.6 pg/ml,with the statistically significant difference (trL10 =3.484,tIL-17 =4.076,P＜0.01).The level of positive group in SLE were 176.5±93.2 pg/ml and 237.2±107.5 pg/ml,and negative group were 16.2±5.7 pg/ml and 12.9 ±8.3 pg/ml,with the statistically significant difference (tIL-10 =6.875,tIL-17 =8.843,P＜0.01).The level of IL-10 was positively correlated with IL-17 level in SLE (r=0.96,P＜0.05).The level of IL-10 and IL-17 in ANA high titer group were 91.8±100.8 pg/ml and 135.5±140.9 pg/ml,ANA low titer group were 44.5±76.7 pg/ml and 54.4±98.5 pg/ml,with the statistically significant difference (tIL10 =2.215,tIL-17 =2.345,P＜0.05).The level of IL-10 and IL-17 in anti ds-DNA antibody positive group were 87.1 ± 101.1 pg/ml and 122.4 ± 137.1 pg/ml,negative group were 27.4± 50.1 pg/ml and 28.6 ± 61.6 pg/ml,with the statistically significant difference (tIL-10 =2.904,tIL-17 =3.443,P＜0.05).Conclusion The levels of IL-10 and IL-17 were significantly increased and there was positive correlation.It was explained that the anti-inflammatory and pro-inflammatory response existing at the same time in SLE patients and Treg cell and Th17 cell may also play an important role in the occurrence and development in SLE.

Objective To investigate the expressions of E-cadherin (E-cad)in arterial chemoembolization interventional therapied bladder carcinoma.Methods The expressions of E-cad in bladder tumor tissues of30 non-muscle-invasive bladder carcinoma treated with preoperative interventional chemotherapy and 20 invasive bladder carcinoma treated with surgical were measured by streptavi-din-peroxidase immunohisto chemical method.The changes of E-cad expression in bladder carcinoma before and after interventional treatment were analyzed.Results The averaged normal expressions rate of E-cad in non-muscle-invasive and muscle invasive bladder carcinoma was 70.0% (21/30),25.0% (5/20)respectively.The averaged normal expressions rate of E-cad after interventional treatment was improved to 90% (27/30),the differences were statistically significant (P <0.05 ).Conclusion The expressions of E-cad in bladder carcinoma had significant relations with pathological grade and clinical stage.The abnormal expressions of E-cad in the mucosal surface, may be associated with inflammation.Interventional treatment can significantly improve the expressions of E-cad of tumor tissue and delay the progress of bladder cancer.

Objective:To analyze the effects of protein intake of varied dietaries on the pregnancy of pregnant women.Methods:The pregnant women who had their medical records in our hospital were included in this study.Their dietary were collected based on the respective review of their food during 24 hours when they visited the Nutrition Clinics between 24 weeks and 28 weeks.The subjects were divided into six groups according to the dietary calories and protein intake:group A (normal calorie and normal protein),group B (normal calorie and low protein),group C (normal calorie and high protein),group D (high calorie and normal protein),group E (high calorie and high protein) and group F (low calorie and low protein).The gestational diabetes mellitus incidence rates,levels of fasting blood glucose (FBG),blood glucose (BG),hemoglobin Alc (HbAlc),and blood lipids,and bone mineral density were compared among six groups.Results:The gestational diabetes mellitus incidence rate,levels of postprandial blood glucose at 1 hour and 2 hour in groups of B,C,D,and E were significantly higher than those in group A (P ＜ 0.05).The levels of triglyceride,total cholesterol,and low density lipoprotein cholesterol were remarkably higher in groups of B,D,and E than those in group A (P ＜ 0.05).The bone mineral density was significantly lower in group A than that in groups of C,E,and F (P ＜ 0.05).Conclusion:The diet with normal calorie and normal protein is the scientific dietary structure.The pattern with high calorie and high protein intake is not recommended for pregnant women.

Wilms tumor(WT) is one of the most common renal malignancies in children.Although several genetic loci such as the WT1,WT2,p53 and ?-catenin genes have been considered to be associated with WT,the causes of the tumor are still unknown.Recently the US researchers have identified a new tumor suppressor gene that is mutated in WT.The biological function of the protein encoded by WTX is yet unknown,however,the gene's location in the X chromosome is of particular interest.This review highlights the current study of the gene mutated in Wilms tumor.

Objective To investigate the value of immunophenotypes in pathologic diagnosis of follicular lymphoma (FL) and the differential diagnosis between neoplastic follicle (NF) and reactive hyperplastic follicle (RHF).Methods 50 cases of FL and 10 cases of RHF as control were studied by clinical data, the expressions of CD20, bcl-2, CD3, CD10, bcl-6, CD21 and Ki-67 were detected by EnVision immunohistochemical method for the immunphenotypical pattern of FL and RHF.Results Among 50 FL cases , there were 24 male cases and 26 female cases, with median age of 50 years old (10-80 years old), including 32 cases (64 ％) involved predominantly neck lymph nodes.The histologic grades were 1-2 in 16 cases (32 ％) and ≥3 in 34 cases (68 ％).CD20, CD10 and bcl-6 were positive in NF tissues of FL, with irregular forms of NF and loss of the mantle area without clear demarcation, and infiltrating to NF, irregular shape and no clear perimeter.The bcl-2 was positive in all of grade 1-2 NF, and it was positive in 68 ％ (23/34) cases and was partial or total negative in 32 ％ (11/34) cases in grade≥3 NF.Those were poorly demarcated contours.The Ki-67 proliferative index of NF were ＜30 ％ in grade 1-2, and ＞30 ％ in grade ≥ 3, with the highest to 90 ％.In FL, the CD21 staining result showed follicular dendritic cells (FDC) network were synchronized with NF, and in some cases of grade ≥ 3, the FDC network were ruptured.In RHF, the CD20 was positive for round or oval nodules with clear demarcation.The CD10, bcl-6 and CD21 were positive and bcl-2 was negative in all germinal center without invasive immunophenotype.The Ki-67 index was high in germinal center, and sometimes polarity may be seen.Conclusion The immunophenotypical differences between FL and RHF include invasive characteristics, loss of follicular mantle area and germinal center pattern in the NF, while the normal follicular immunophenotype in the RHF.

10.3969/j.issn.2095-4344.2013.25.011

ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

Objective To observe the relationship of IL-10 gene single nucleotide polymorphism and the susceptibility to ALL. Methods The bone marrow and peripheral blood samples from 115 ALL patients and 323 healthy controls were collected in Peking University First Hospital and Beijing Dao-pei Hospital from January 2007 to December 2009. The DNA were extracted from all samples. The primers of -819C/T and -592A/C in the promoter region of IL-10 gene were designed for the PCR. The restrictive fragment length polymorphism of IL-10 gene was analyzed by using restrictive enzyme Msl Ⅰ and HpyCH4 Ⅲ.Sequencing was done in part of these samples to confirm the results of PCR. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the ALL patients and healthy controls. Real-time quantitative PCR was performed to detect the EB virus (EBV) infection and the expression of BCR/ABL fusion gene. The differences of genotypes and allele ratio of -819 and -592 sites were analyzed between the positive and negative group. Results The genotype ratios of -819CC, -819TT, - 819CT, -592AA,- 592CC and - 592AC were 14. 8% ( 17/115 ), 45.2% ( 52/115 ), 40. 0% ( 46/115 ), 43.5% ( 50/115 ),16. 5% ( 19/115 ), 40. 0% ( 46/115 ) in ALL patients, and were 9. 9% ( 32/323 ), 16. 4% ( 53/323 ),73.7% ( 238/323 ), 11.8% ( 38/323 ), 15.5% ( 50/323 ), 72. 8% ( 235/323 ) in the healthy controls,respectively. The genotypes of -819 and -592 sites had statistically significant differences between the two groups(x2 values were 46.000 and 54.550, all P ＜ 0. 05 ). The allele ratio of -819T and -592A were (65.2%, 150/230) and (63.5%, 146/230) in ALL patients, while they were 53.5% (344/646) and 48. 1% (311/646)in the healthy controls. There were statistically significant differences between the two groups (x2 values were 9. 877 and 15.986, all P ＜ 0. 05 ). The EBV DNA were detected in 42 ALL patients,among which 22 were positive and 20 were negative. The genotype ratios of -819CC, -819TT, -819CT,-592AA, - 592CC, - 592AC in EBV positive group were 9. 1% ( 2/22 ), 40. 9% ( 9/22 ), 50. 0%(11/22) ,31.8% ( 7/22 ), 13.6% ( 3/22 ), 54. 5% ( 12/22 ), while they were 35.0% ( 7/20 ), 45.0%(9/20) ,20. 0% (4/20) ,35.0% (7/20) ,45.0% (9/20) ,20. 0% (4/20) in the EBV negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups( all P ＞ 0. 05 ).The BCR/ABL fusion gene were detected in 36 ALL patients, among which 20 were positive and 16 were negative. The genotype ratios of - 819CC, - 819TT, - 819CT, - 592AA, - 592CC, - 592AC in BCR/ABL positive group were 0% (0/20) ,45.0% (9/20) ,55.0% ( 11/20), 45. 0% (9/20) ,5.0% (1/20) ,50. 0%( 10/20), while they were 18. 8% ( 3/16 ), 50. 0% ( 8/16), 31.3% ( 5/16 ), 50. 0% ( 8/16 ), 18. 8%(3/16), 31.3 % (5/16)in the BCR/ABL negative group. The genotypes of -819 and -592 sites showed no statistical differences between the two groups ( all P ＞ 0. 05 ). Conclusion The population with - 819TT and - 592AA genotype of IL-10 gene shows susceptibility to ALL.

Objective To analyze nucleophosmin (NPM1) gene mutations in exon 12 in patients with acute myeloid leukemia (AML) and evaluate the clinical appliance of three methods which are frequently used for detecting gene mutation. Methods Genomic DNA from bone marrow of 54 AML patients was detected by PCR for NPM1 exon 12 and screened by PCR-capillary electrophoresis, denature high-performance liquid chromatography (DHPLC) and direct sequencing separately. FLT3-ITD (FMS-like tyrosine kinease internal tandem duplication) was detected by agarose gel electrophoresis and PCR-capillary electrophoresis. Results Seven AML sample harbored NPM1 gene mutations. Five of them were the most common mutation, known as type A (an insertion of a TCTG tetranucleotide at position 960 bp). One of them was type D (an insertion of a CCTG tetranuclectide at position 960 bp). The new variant was a deletion of a TGGCAGTG sequence at 958 bp and insertion of a GCCCGCGGTTTA sequence instead. The detection ratio of the three methods was all 100% and capillary electrophoresis was more rapid, reliable and easier than the other two methods. Moreover it could detect FLT3-ITD simultaneously. The resolving power of DHPLC was affected by many factors. The direct sequencing method was tedious and the heterozygous sequence might be misread. Conclusions There is a new mutation at position 958 bp with a 12-nucleotide insertion and substitution. PCR-capillary electrophoresis is convenient to screen NPM1 mutations of AML in clinical practice.

Objective To investigate differences of ctinicopathologic features in breast cancer patients with or without Type 2 Diabetes Mellitus(DM).Methods The general conditions and elinicopathologic features of the hospitalized breast cancer patients with or without type 2 diabetes were analyzed using a case-control study.Results The average age,fasting blood-glucose,BMI and TG were(58.4 ± 7.80),(8.15 ± 2.80)mmol/L,(27.72 ± 3.47)mmol/L and(2.36 ± 1.18)mmol/L in patients with DM,and(51.6 ± 9.90),(5.13 ±0.63)mmol/L,(24.15 ± 4.95)mmol/L and(1.32 ± 0.59)mmol/L in patients without DM.There were significantdifference between the two groups(t =2.968,P =0.004; t =5.757,P ＜ 0.001 ; t =3.235,P =0.002; t =4.330,P ＜0.001,respectively).HDL-C in patients with DM was(1.39 +0.20)mmol/L,which was significantly lower than that of(1.50 ± 0.24)mmol/L in patients without DM(t =2.000,P =0.05).TC and LDL-C was(4.89 ± 1.16)mmol/L and(3.02 ±0.90)mmol/L in patients with DM,which were not significantly different with those of(4.79 ±0.85)mmol/L and(2.97 +0.61)mmol/L in patients without DM(t =0.396,P =0.693,and t =0.255,P =0.800,respectively).More patients were in the menopausal status in breast cancer patients with Type 2 DM compared to the other group(x2 =11.835,P =0.001).The expression of Her-2 was 76.7％(23/30)in breast cancer patients with Type 2 DM,which was significantly higher than that of 50.8％(33/65)in patients without DM(x2 =5.689,P =0.017).Conclusion The average age was higher in breast cancer patients with Type 2 DM and most of them were in their menopausal status,furthermore the higher body mass index and worse prognosis were observed in this group,so the breast cancer patients with diabetes should choose the more reasonable treatment.