Objective To investigate the relationship between the expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF) and the angiogenesis in human malignant melanoma (MM). Methods Tissue sections from 31 MM patients were examined. The protein expression of eNOS and VEGF was detected using immunohistochemistry technique and morphological quantitative analysis. Microvessel density (MVD) was also counted by immunostaining with anti-FⅧRAg antibody. Results (1)The positive staining of eNOS and VEGF was detected in 71.4% (24 cases) and 83.9% (26 cases) of MM respectively. As negative control there was no staining in the nevus pigmentosus tissue sections. (2)There was a positive correlation between the expression of eNOS and VEGF in MM. (3)The positive correlation was also found between the MVD and the expression of eNOS and VEGF in MM. The number of MVD was higher in MM than that in nevus pigmentosus tissue. (4)The lymphatic metastasis was not correlated with the expression of eNOS and VEGF. Conclusion The number of the MVD increases along with the high expression of eNOS and VEGF in MM.

Objective To explore the relationship between expression of p27 and cyclin D1 protein in keratinocyte－ derived tumors. Methods Expression of p27 and cyclin D1 protein was determined in various keratinocyte－ derived tumors by immunohistochemistry techniques. Results Significantly higher expression of p27 was found in both Bowen′ s disease and actinic keratosis(AK) than that in basal cell carcinoma(BCC) and squamous cell carcinoma(SCC)（ P

Objective To evaluate the specificity,sensitivity and clinical significances of direct antiglobulin test of bone marrow monouclear cells (BMMNC-Coombs test) and Flow Cytometry (FACS).Methods Autoantibodies on bone marrow mononuclear cells of 270 patients with immune and 471cases with primary or secondary anaemia and 19 normal controls were measured by BMMNC-Coombs test,and a part of IRP patients with negative BMMNC-Coombs test were retested by FACS.Results In 270 immune hemocytopenia patients,there were 163 (60.37%) with positive BMMNC -Coombs test.The positive rate of patients with immunorelated pancytopenia (IRP) was upmost and accounted for 67.2%.AIHA 63.6%,ITP 57.1%,Evans syndrome 43.2%.Both 471 patients with primary or secondary anaemia and 19 normal persons were all negative.In BMMNC-Coombs positive cases,there were mainly three types including IgG,IgG+C_3,IgG+IgM+IgA+C_3,they accounted for 30.4%,26.4%,20.8%.61 1RP with negative BMMNC-Coombs test were teased again for their BMMNC autoantibodies by FACS.83.6% of them were found to be positive with autoantibodies on BMMNC.The positive rate of autoantibodies on hematopoietic stem cells of IRP patients was 94.1%.it was higher than positive rate on any other types of bone marrow hematopoietic cells.Conclusion the sensitivity and specificity of BMMNC-Coombs test for diagnosis of autoimmune hemocytopenia was high,but the sensi- tivity of FACS assay was higher than that of BMMNC-Coombs test for detecting autoantibodies.Also had IRP,AIHA, ITP and Evans syndrome patients autoantibodies on bone marrow hematopoietic cells,The difference between IRP and other immune hemocytopenias was that IRP patients did have autoantibodies only on BMMNC.

In order to explore the effects of Panax notoginoside (PNS) on the expression of transforming growth factor β1 (TGF-β1) and Smad-7 in renal tissues of diabetes, a rat model of diabetic nephropathy was set up by intravenous injection of streptozotocin (STZ). Wistar rats were randomly divided into normal group, diabetic control group, group treated by PNS at low-dosage (PL), group treated by PNS at high-dosage (PH) and group treated by catopril (C), respectively. Fasting blood glucose (FBG), renal index, endogenous creatinine clearance rate (C(Cr)) and urinary albumin (UAlb) in 24 h were examined after 6 weeks. Meanwhile, the expressions of TGF-β1 and Smad7 in renal tissues were immunohistochemically dectected. At the end of the sixth week, FBG, renal index, C(cr), UAlb were all elevated significantly in control group (P<0.01). The expression of TGF-β1 protein was increased while Smad7 protein decreased in renal tissue (P<0.01). However, the treatment with PNS reversed the aforementioned changes in renal tissues of diabetic rats. These results indicate that PNS possess a protective effect on the kidney of diabetic rats and it might protect kidney by inhibiting the expression of TGF-β1 protein and enhancing the expression of Smad7 protein.

Objectives To study the effects of pU- vascular endothelial growth factor (VEGF)- short interfering RNA (siRNA) on the formation and apoptosis of malignant melanoma in models of nude mice and its mechanism. Methods The siRNA eukaryotic expression vector for VEGF was constructed, then transfected into A375 (a human malignant melanoma cell line) by electroporation. The nude mice models of malignant melanoma were constructed. The protein expression of VEGF and factor Ⅷ related antigen (FⅧRAg) specific for vascular endothelial cells was detected by immunohistochemical technique and morphological quantitative analysis. Microvessel density (MVD) was counted based on the endothelial cells positively stained with anti-FⅧRAg antibody. The apoptosis of the neoplasms in nude mice was quantitatively determined by TdT mediated dUTP nick end labeling (TUNEL). Results Both the number and the growth speed of the neoplasm formation were lower in the experimental group than those in the controls (both P

Objective To analyze the pathogenesis,initially diagnosed symptoms and clinical manifestations of children with chronic kidney disease (CKD) at stage 2 to 5.Methods The data of 108 children who were hospitalized in Children's Hospital Affiliated to Capital Institute of Pediatrics from September 2007 to April 2016 with CKD stage 2 to 5 were retrospectively analyzed.The etiologies,clinical manifestations and examinations were summarized,and the clinical manifestations were compared between the congenital hereditary urinary diseases group and the acquired urinary diseases group.Results (1) In the 108 cases collected,66 cases were male,42 cases were female,aged from 3 months to 15 years and 1 month old.Twenty-four cases were diagnosed at stage 2,26 cases at stage 3,35 cases at stage 4,and 23 cases at stage 5.(2) Twenty-eight kinds of illness were involved in the cause of CKD.Among them,57 cases (52.8％) had congenital anomalies of the kidney and urinary tract,5 cases(4.6％) had hereditary kidney diseases,41 cases (38.0％) had other primary or secondary kidney diseases,and in 5 cases (4.6％) the causes of disease were unknown.(3) For the initially diagnosed symptoms,29 cases(26.9％) were due to complaints associated with kidney disease,36 cases (33.3％) were of other outside kidney symptoms,and 43 cases (39.8 ％) were of negative symptoms.The results of urinary ultrasound were abnormal in 79 cases(73.1％) and 87 cases(80.6％) showed abnormality in urinary analysis.There were 105 cases (97.2％) with abnormal manifestations either in urinary tract ultrasound or in urinary analysis.(4)The ages on diagnosis as CKD in children with congenital hereditary urinary diseases(5.89 years old) were younger than that of children with acquired urinary diseases (9.20 years old),and the difference was significant(Z =-3.434,P =0.001).The frequency of cases with short stature or lower-weight in group of congenital hereditary urinary diseases[66.1％ (41/622 cases),64.5％ (40/62 cases)] were significantly higher than those of the acquired urinary diseases group[43.9％ (18/41cases),43.9％ (18/41 cases)],and the differences were statistically significant(x2 =4.983,4.263,P =0.026,0.039).Conclusions The causes of CKD are complicated,and the congenital anomalies of kidney and urinary tract are the major causes of CKD at stage 2 to 5 in the cases.The initially diagnosed symptoms of CKD are insidious and atypical.The children with congenital hereditary urinary diseases tend to have more serious growth retardation.Urinary analysis and ultrasound may have an important significance for early diagnosis of CKD in children.

As a traditional Chinese medicine,pheretima is widely used for thousands of years in China. According to the studies in recent years,the pharmacological effects and clinical researches of pheretima were summarized based on the introduction of chemical constituents, which could provide information for the further study.

Current reform of higher medical education focuses on curriculum integration and corresponding reconstruction of teaching system. Chongqing Medical University has carried out the reform of medical personnel training mode from basic to clinical, which has achieved certain results. Three aspects including teaching content, teaching methods and appraisal system of the urogenital system curriculum integration are introduced in the paper. In the teaching content, the parts related to urogenital system in basic subjects and clinical subjects are extracted and integrated. In the process of teaching implementation, a teaching team is set up across departments and multidisciplinary joint teaching is carried out. In order to arouse the enthusiasm of students, inspire and cultivate students' scientific research thinking by improving teaching methods, several teaching methods such as case introduction teaching method, question discussion and debate meeting are used. Scientific evaluation system is used to observe and record the whole process of students' learning. Meanwhile, some existing problems and solutions of curriculum integration are discussed in the paper to provide references for peers in relevant colleges and universities.

Objective To investigate the effect of recombinant murine interleukin-23(rIL-23)on systemic candidiasis in a murine model.Methods A cyclophosphamide-induced immunosuppressed murine model of systemic candidiasis was established.The mice were divided into control group and rIL-23 treatment group.Colony forming units(CFU)of the kidney and spleen were determined by using plating dilution method.The histopathological changes and degree of infection of the kidney and spleen were graded.Meanwhile,the levels of interferon gamma(IFN-γ)in the spleen were measured by enzyme-linked immunosorbent assay.Results On the 2nd,3rd and 7th day after Candida albicans infection the number of CFU of the fungi in the kidney in the control group was significantly greater than that in rIL-23 treatment group(P＜0.01).The number of CFU of the fungi on the 2nd,3rd and 7th day after Candida albicans infection in the spleen in control group was also greater than that in rIL-23 treatment group,but without statistically significant difference(P＞0.05).The scores of histopathological changes in the kidney in rIL-23 treatment group were lower than those in control group(P＜0.01),and the degree of infection was milder in rIL-23 treatment group.The scores of histopathological changes in the spleen in rIL-23 treatment group were also lower than those in control group,but without statistically significant difference(P＞0.05).The levels of IFN-γ in the spleen on the 2nd,3rd and 7th day after infection in rIL-23 treatment group were significantly higher than those in control group(P＜0.01).Conclusion rIL-23 has protective effect on murine systemic candidiasis.

Objective To construct an eukaryotic expression vector for TAP genes fused with enhanced green fluorescent protein(EGFP)gene,and to analyze the expression and subcellular localization of the fusion protein in A375 human malignant melanoma cells transfected with the eukaryotic expression vector.Methods A375 cells were cotransfected with the combination of plasmid(P)TAP1-EGFP or pTAP2-EGFP and pDsRed2-endoplasmic reticulum(ER),or with pEGFP-TAP1 and-TAP2,or monotransfected with pTAP1-EGFP or pTAP2-EGFP alone.The monoclonal A375 cells stably expressing TAP were obtained by G418 selection.Then.the distribution and expression of fusion proteins were assessed in A375 cells by using fluorescence microscopy and Western blot,respectively.Flow cytometry was used to measure the expression of HLA class Ⅰ on A375 cells.Results Transfection of A375 cells with pTAP1-EGFP or pTAP1-EGFP and pTAP2-EGFP significantly increased the expression of TAP 1 and TAP 2 in as well as HLA class Ⅰ antigen on A375 cells.The green fluorescence of TAP1-EGFP and TAP2-EGFP overlapped with the red fluorescence of ER marker in cotransfected cells.indicating that TAP was localized subcellularly on the ER.Conclusions The expression vector for TAP-EGFP fusion gene has been constructed cuccessfully and expressed in A375 cells,and the expressed fusion protein is subcelluiarly localized to ER.This study will provide a basis for the research into subsequent immune response following induction of TAP expression.