0.05). In the second and third years, average scores were decreased ( P

As a member of regenerating islet-derived family,regenerating islet-derived type Ⅳ(RegⅣ)is involved in cell proliferation and differentiation. RegⅣ is closely related to the occurrence and develop-ment of tumor,tumor cell proliferation and invasion and anti-apoptosis,and it may be a potential molecular tar-get of targeting therapy in cancer.

Objective To detect the level of Tnl and T.2 type cytokines in the patients with lym-phoma in order to find out the laboratory evidence of tumor immunotherapy. Methods The levels of serum IFN-γ, TNF-α, IL-2, IL-4, IL-5 and IL-10 were measured by cytometric bead array (CBA) in 92 patients with lymphoma, and 70 normal sera as control. Results The levels of TH1 type cytokincs in 92 patients with lymphoma were: IFN-γ (34.26 ± 33.48) pg/ml, TNF-α (8.17 ± 10. 09) pg/ml, IL-2 (3.74 ±1.72) pg/ml; and the levels of TH2 type cytokines were: IL-10 (6. 28±8.56) pg/ml, IL-5 (3.53 ±3.20) pg/ml, IL-4 (6.22±7.13) pg/ml. The levels of TH1 and TH2 cytokines in lymphoma patients were significantly higher than those in controls(P ＜ 0.01) except TNF-α. And the rate of IL-2/IL-4 was signifi-cantly decreased in lymphoma patients(P ＜0.01). The level of IL-10 in Ⅲ/Ⅳ stage lymphoma patients was much higher than that in Ⅰ/Ⅱ stage patients(P ＜0.01). The level of IFN-γ/was significantly decreased in aged patients with lymphoma (P ＜0. 05). Conclusion TH 1/TH2 is imbalance in lymphoma, which may provide clinical index for the evaluation of progrossien and prognosis. In the lymphoma patients TH1/TH2 shifts to TH2, which may be the mechanism of tumor arising and transferring by immune escape from immu-nosurveillance.

AIM:To establish some HPLC-digitized fingerprint spectrum(HPLC-DFPS)in Liuwei Dihuang Dripping Pill preparation and to apply it to identification of medicinal materials.METHODS:Based on loganin and paeonoside contents adopted as qualitative and quantitative factors,relative retention value as creteria,RPHPLC was carried out on inensil ODS:column,mobile phase consisted of methanol-water(0.2% formyl acid),detection wavelength was set at 240 nm.RESULTS:Using above optimum chromatograph conditions,HPLC-DFPS of relative medicinal materials andLiwei Dihuang Dripping Pillpreparation were established.By comparison a group of characteristic peaks could be used to identify medicinal materials.CONCLUSION:This result indicates that it is practical to use HPLC-DFPS for identification of Liawei Dihuang Dripping Pill.

AIM:To establish some HPLC-digitized fingerprint spectrum(HPLC-DFPS)in Liuwei Dihuang Dripping Pill preparation and to apply it to identification of medicinal materials.METHODS:Based on loganin and paeonoside contents adopted as qualitative and quantitative factors,relative retention value as creteria,RP-HPLC was carried out on inertsil ODS:column,mobile phase consisted of methanol-water(0.2% formyl acid),detection wavelength was set at 240 nm.RESULTS:Using above optimum chromatograph conditions,HPLC-DFPS of relative medicinal materials and "Liwei Dihuang Dripping Pill" preparation were established.By comparison a group of characteristic peaks could be used to identify medicinal materials.CONCLUSION:This result indicates that it is practical to use HPLC-DFPS for identification of Liuwei Dihuang Dripping Pill.

Background and purpose:The prognostic role of human epidermal growth factor receptor 2 (HER-2) remains controversial in patients with non-small cell lung cancer (NSCLC). This study aimed to evaluate the expression of HER-2 and its prognostic implication in patients with NSCLC.Methods:Four hundred and twenty-ifve NSCLC patients’ specimens were obtained from Zhejiang Cancer Hospital. HER-2 protein expression was determined by immunohistochemistry (IHC). The relationship between HER-2 and clinicopathological parameters and patients’ prognosis was analyzed by Chi-square test and COX proportional hazards regression model.Results:Eighty-three patients were positive expression of HER-2 (83/425, 19.5%). The expression of HER-2 was closely related with histological type (P=0.051), most patients with HER-2 positive were adenocarcinoma, then were squamous carcinoma or other histological types. However, HER-2 was not an independent prognostic indicator for the overall survival of patients with NSCLC.Conclusion:We found that the expression of HER-2 was closely connected with histological type, but it was not a predictive marker for the prognosis in patients with NSCLC.

Objective To investigate the relationship of the metastasis-associated genes and its copy numbers variation in the highly metastatic human epithelial ovarian cancer cell line HO-8910PM. Methods The differentially expressed genes and its copy number variation between HO-8910PM cell line and normal ovarian tissues was detected by human genome UI33A 2.0 gene chip and human mapping 10K array 2.0 gene chip, and the data was analyzed by bioinformatics. Some of metastasis-associated genes were validated the results of single nucleotide polymorphism (SNP) and cDNA chips by fluorescence in situ hybridization (FISH) and real-time quantitative PCR. Results Integrate analysis of two gene chips data showed that there were 385 differentially expressed genes in the same and 379 SNP positional point (6 of them, included 2 genes) between HO-8910PM cell line and normal ovarian tissues, these copy number amplification of 379 SNP positional point of chromosome were ≥3, which had 240, deletion ≤ 1 had 139. Chromosome location analysis showed that there were 385 differentially expressed genes located at all chromosomes, and 261 of them ( 67.8%, 261/385 ) located at 10 chromosomes, included that 34 (8.8% ), 33 ( 8.6% ), 28 (7.3%), 27 (7.0%), 25 (6.5%), 24 (6.2%) of them located at chromosome 3, 2, 9, 10, 1 and 11 respectively, and 23 (6.0%) of them at chromosome 6 and 12 each, 22 (5.7%) of them at chromosome 4 and 5 each. For the function of differentially expressed genes, the results showed that 99 (25.7% ) genes belonged to the family of enzymes and their regulators, 54 ( 14.0% ) genes associated with signal transduction, 50 (13.0%) genes associated with nucleic acid binding, and 36 (9.4%) genes associated with protein binding. Conclusion We have demonstrated that there are 4 kinds of differentially expressed genes related to metastasis of ovarian cancer, which belonged to the families enzyme and its regulator, nucleic acid binding, signal transduction and protein binding, and located at chromosome 1, 2, 3, 4, 5, 6, 9, 10, 11 and 12.

Objective To evaluate the feasibility of apply Ion Proton semiconductor sequencing platform in non-invasive prenatal genetic diagnosis .Methods Totally 1 000 pregnant women with a singleton pregnancy of 12-32 weeks gestation were selected from the Third affiliated Hospital of Zhengzhou University from Jan to Dec 2013.Using noninvasive prenatal genetic diagnosis based on Ion Proton semiconductor sequencing platform to study their cffDNA .In parallel, 72 pregnant women received invasive prenatal diagnosis by traditional chromosomal analysis with amniocentesis chorionic villus sampling .Results It′s shown that 18 out of 1 000 (1.8%) pregnant women underwent the noninvasive prenatal genetic testing had a high risk for aneuploid chromosomes , including 7 cases of 21-trisomy, 4 cases of 18-trisomy, 2 cases of 13-trisomy, 4 cases of sex chromosomal abnormality , and 1 case of 15-trisomy.It demonstrated that the rate and accuracy of fetal 21-trisomy, 13-trisomy and 18-trisomy by non-invasive prenatal genetic testing were both 100%without misdiagnosis , the rate of detection for sex chromosomal abnormality was 2/2 with a false positive rate of 1/3.However, the 15-trisomy predicted by the non-invasive prenatal diagnosis in a woman was finally proved to be a false positive .Based on the results by karyotyping (55/55) as well as follow-ups (493/493), the specificity of the non-invasive prenatal diagnosis for detection of 21-trisomy, 18-trisomy and 13-trisomy was 100%.One Ion PITM chip could detect 12 to 15 samples in 1.5 h and the whole process of noninvasive detection could be completed in 1 to 1.5 days.Conclusions The non-invasive prenatal diagnosis by Ion Proton semiconductor sequencing platform could provide fast and accurate detection of fetal aneuploidy .The benchtop high-throughput sequencing platform has laid the foundation for the independent application in clinical settings for fetal aneuploidy detection .

Objective To compare the effect of transplant islets between the subcutaneous inguinal white adipose tissues and renal capsule in the treatment of type 1 diabetes mellitus in mouse models. Methods The mice with type 1 diabetes mellitus undergoing islet transplantation were divided into the white adipose group (n=10) and renal capsule group (n=10). The islets were isolated, purified and transplanted to the subcutaneous white adipose tissues of inguinal region and renal capsule. The random blood glucose level and glucose tolerance function of the recipient mice in two groups were continuously monitored after operation. Islet grafts of the surviving recipient mice were harvested at postoperative 100 d for histopathological examination. Results In the white adipose group, the blood glucose levels of 6 recipient mice were restored to normal at 1 month after transplantation, whereas the blood glucose levels of the other 4 recipient mice were high, which died before the end of monitoring. In the renal capsule group, the blood glucose levels of 10 recipient mice returned to normal within 10 d after transplantation. Islet grafts of the recipient mice in two groups could lower the blood glucose levels, whereas the islet grafts in the white adipose group required a longer time to exert the effect. The glucose tolerance function of the mice in the renal capsule group was significantly better than that of those in white adipose group (P < 0.05). Histopathological examination demonstrated that the insulin of the islet grafts was normally expressed in two groups. Conclusions The islets transplanted into the subcutaneous white adipose tissues of inguinal region can play an effective role in regulating the changes of blood glucose level. Although the blood glucose-lowering function is slightly weaker than that of the islets graft in the renal capsule, it has multiple advantages resembling the ideal islet transplantation sites, which is a promising replacement site for islet transplantation.

Objective To investigate the isolation and culture of porcine bone marrow mesenchymal stem cell (BMSC) with α-1, 3-galactosyltransferase (GGTA1) gene knockout (GTKO), GTKO/ human CD46 (hCD46) insertion and cytidine monopho-N-acetylneuraminic acid hydroxylase (CMAH)/GGTA1 gene knockout (Neu5GC/Gal), and the protective effect of co-culture with porcine islets on islet cells. Methods Bone marrow was extracted from different transgenic pigs modified with GTKO, GTKO/hCD46 and Neu5GC/Gal. Porcine BMSC were isolated by the whole bone marrow adherent method and then cultured. The morphology of BMSC was observed and the surface markers of BMSC were identified by flow cytometry. Meantime, the multi-directional differentiation induced by BMSC was observed, and the labeling and tracing of BMSC were realized by green fluorescent protein (GFP) transfection. The porcine BMSC transfected with GFP were co-cultured with porcine islet cells. Morphological changes of porcine islet cells were observed, and compared with those in the porcine islet cell alone culture group. Results BMSC derived from pigs were spindle-shaped in vitro, expressing biomarkers of CD29, CD44, CD73, CD90, CD105 and CD166 rather than CD34 and CD45. These cells were able to differentiate into adipocytes, osteoblasts and chondrocytes. Porcine BMSC with GFP transfection could be labeled and traced, which could be stably expressed in the daughter cells after cell division. Porcine BMSC exerted certain protective effect on islet cells. Conclusions GFP-labeled porcine BMSC modified with GTKO, GTKO/hCD46 and Neu5GC/Gal are successfully established, which exert certain protective effect upon islet cells.