Objective To inveatigate the expression and clinical significance of microRNA‐204(miR‐204) in human renal clear cell carcinoma(RCCC) .Methods 65 cases of resected RCCC tissue and corresponding normal tumor‐adjacent tissue were col‐lected and detected the expression level of miR‐204 by using qRT‐PCR .The correlation between the miR‐204 expression level and clilicalpathological features was statistically analyzed .The artificial miR‐204 mimics were adopted to stranfect into Caki‐1 cells ,then the Western blot was used to detect the expression of Bcl‐2 a as the downstream potential targets of miR‐204 and SIRT1 mRNA and protein .Results The expression level of miR‐204 in the RCCC tissue was significantly lowere than that in the normal tumor‐adjacent tissues (P<0 .05);the low expression of miR‐204 was significantly associated with the tumor size (>3 cm ,P<0 .05) , lymph node metastasis(P<0 .05) and advanced TNM stage(Ⅲ + Ⅳ ,P<0 .05) .Both the mRNA and protin expression in RCCC Caki‐1 cells were down‐regulated after transfection .Conclusion The expression of miR‐204 is down-regulated in the RCCC tissue and is related to the malignant clinicopathological features ,and miR‐204 may suppress the RCCC genesis and development through down‐regulating the expression of Bcl‐2 and SIRT1 .

Objective To explore effect of preoperative neutrophil/lymphocyte ratio (NLR) on the prognosis of patients with prostate cancer.Methods A total of 51 cases died included in our hospital as the died group, also included 124 patients survival patients served as survival group.Preoperative total prostate specific antigen (tPSA), free prostate specific antigen (fPSA), albumin, white blood cell (WBC), neutrophil count, lymphocyte count, Gleason score, and NLR were collected.Receiver operating characteristic (ROC) curve was drawn.The function of the index on prognosis was evaluated.Results The average age was 74.0 (9) years in the died group, and 73.6 (11.0) years in the survival group.Gleason score was 8 ～ 10.The patients were followed up for 10 ～ 36 months with a median follow-up time of 23 months.No significant differences in albumin and Gleason score were found between death and survival groups (P ＞0.05).However, the other major indexes had significant differences between two groups (P ＜ 0.01).NLR area under curve (AUC) was 0.835.The NLR boundary value was 2.84, which was higher than the critical value of patients with poor prognosis.Conclusions NLR can predict the prognosis of patients with malignant prostate cancer as auxiliary index.Patients with NLR ＞ 2.83 had poor prognosis.

Objective To study the effects of anti-keratin autoantibody (AK auto Ab) on apoptosis of human keratinocytes. Methods Light and electron microscopy were used to observe morphological changes, flow cytometry (FCM) to analyze cell cycle, and electrophoresis to analyze DNA profiles of cultured keratinocytes. Results Typical morphological changes with apoptotic characteristics such as karyopyknosis, chromatin agglutination and apoptosis bodies were found. The subdiploid peaks due to apoptosis were also found in cell cycle analyses. DNA electrophoresis of keratinocytes showed characteristic ladder. Conclusions AK auto Ab could induce apoptosis in cultured human keratinocytes.

Objective To study the effect of cytokine-induced killer(CIK) cells immunotherapy on immunological func-tion of patients with metastatic renal cell carcinoma.Methods A total of 70 patients with metastatic renal cell carcino-ma were randomly divided into 2 groups, 35 cases of the treatment group were added with CIK cells, while the control group was given IL-2 and IFN-α, detected the immunological function of 70 cases including the levels of CD3+, CD4+, CD8+T cells, ratio of CD4+/CD8+, one week before therapy of CIK cells and one month week after treatment. Results The levels of CD3+、CD4+cells, ratio of CD4+/CD8+in peripheral blood of patients with renal cancer of two groups were much higher than those before therapy (P<0.05), the levels CD8+ was lower after treatment, there were significant changes (P<0.05). Conclusion CIK cells immunotherapy can enhance the immunological function of patients with meta-static renal cell carcinoma.

ObjectiveTo determine the pharmacodynamic substance basis of Epimedii Folium（EF） and Epimedii Wushanensis Folium（EWF） in promoting osteogenic differentiation， and to establish a method to analyze the material basis of Chinese materia medica based on the correlation between chemical fingerprint and cellular metabolomics. MethodThe chemical fingerprints of 15 batches of EF with 4 species and 3 batches of EWF were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， and partial least squares-discriminant analysis（PLS-DA） was used to analyze the peak areas of chemical fingerprints of samples. The effects of different samples on proliferative activity of MC3T3-E1 osteoblast precursors， as well as the activity of alkaline phosphatase（ALP） in osteoblasts were detected by cell counting kit-8（CCK-8） and enzyme-linked immunosorbent assay（ELISA）. At the same time， UPLC-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was used to analyze the effects of different samples on the metabolomics of MC3T3-E1 cells， then metabolic peak table of osteogenic differentiation cells was constructed， and pharmacodynamic index mean Y₀ was introduced into the peak table. PLS was used to calculate mean Y₀ of each group， and the mean Y₀ was added to the peak table of chemical fingerprint to construct the correlation between chemical fingerprint and cell metabolome， the pharmacodynamic components of EF and EWF that promote bone differentiation were screened according to variable importance in the projection（VIP） value>1. The pharmacodynamic effects of EF and EWF were evaluated according to the mean Y₀ of each group. ResultThe chemical fingerprints of EF with different origins and EWF were completely separated. Compared with the blank group， the activity of MC3T3-E1 cells in EF and EWF groups was significantly increased， the activity of ALP in the Epimedium brevicornu（Gansu province）， E. koreanum and E. pubescens groups was significantly increased（P<0.05）. The results of cell metabolomics showed that the blank group and the model group had an obvious trend of separation. EF with different origins and EWF had different distance from the model group， indicating that EF with different origins and EWF had different effect on promoting osteogenic differentiation. Chemical fingerprint-cell metabolomics integration analysis screened 9 components closely related to the efficacy of EF and EWF， including diphylloside B， epimedin C， icariin， baohuoside Ⅰ， yinyanghuo B， β-anhydroicaritin， magnoflorine， cryptochlorogenic acid and quercetin. E. koreanum had the strongest effect on promoting osteogenic differentiation. ConclusionThis study determined that the material basis of EF and EWF promoting osteogenic differentiation were mostly flavonoids， alkaloids and organic acids， which provided ideas and methods for the screening of pharmacodynamic components and the prediction of therapeutic effect of Chinese materia medica.