BACKGROUND: Talomerase activity inhibitor inhibits or kills renal carcinoma cells, and also affects stem cells that play importan roles in occurrence and development of renal carcinoma.OBJECTIVE: To observe renal carcinoma stem cell surface marker CD133 and telomerase activity expression in serum-free suspension culture, and to compare with renal carcinoma cells in serum suspension culture.DESIGN, TIME AND SETTING: The in vitro cytological study was performed at the Jiangsu University from June 2008 to Februar 2009.WIATERIALS: Fresh normal renal tissue surrounding renal carcinoma was obtained from Affiliated Hospital, Jiangsu University.Renal carcinoma stem cell line OS-RC-2 was supplied by Cell Bank, Chinese Academy of Sciences Shanghai Branch.METHODS: OS-RC-2 in logarithmic phase, digested by trypsin, and centrifuged. Supematant was removed. OS-RC-2 cell line in serum-free DMEM/F12 supplemented with epidermal growth factor and basic fibroblast growth factor was incubated at 2×105/L in 5% CO2 incubator at 37℃. Renal carcinoma cultured in serum and normal renal tissue served as controls.MAIN OUTCOME MEASURES: Cell growth was observed under an inverted microscope. Expression of CD133 and CD34 was detected using flow cytometry. Reel-time quantitative TRAP assay was applied to evaluate telomerase activity in renal carcinoma stem cells.RESULTS: After incubated in serum-free medium, renal carcinoma stem cells were round and suspended. Two days later, cell mass generated. Each cell mass contained 3-8 cells, with strong refraction. Seven days later, cell mass became more, presented big body that was regular, round or elliptical. CD133+CD34- rate in renal carcinoma stern cell mass was significantly greater in serum-free suspension culture compared with in serum suspension culture. CD133 and CD34 expression was not determined in normal renal tissue. There were significant differences among groups (F=328.25, P < 0.05). Telomerase activity was greater in renal carcinoma stem cells and renal carcinoma cells compared with normal renal ceils (F=-278.74, P < 0.05). No significant difference was detected between renal carcinoma stem cells and renal carcinoma cells.CONCLUSION: Compared with serum cultured renal carcinoma cells, serum-free cultured renal carcinoma cell surface marker CD133 presents high expression. Moreover, talomerase activity is high in renal carcinoma stem cells and renal carcinoma cells compared with normal renal tissue.

Objective To further clarify the pathogenesis of different types of Kaposi′s sarcoma (KS) by measuring the protein expressions of caspase-3 and E-cad in tumor tissues of Xinjiang Uygur patients with acqured immunodeficiency syndrome (AIDS)-Kaposi′s sarcoma (KS) and classical KS.Methods From July 2011 to October 2014, 38 patients with KS at the First Affiliated Hospital of Xinjiang Medical University and Urumqi Infectious Disease Hospital were enrolled, among whom 28 were male and 10 were female, and all of them were uygur.Immunohistochemical and Western blot methods were used to detect the expressions of caspase-3 and E-cad proteins in 22 cases of AIDS-KS patients and 16 cases of classic KS.The quantitative data of normal distribution were analyzed by t test, while count data were compared with χ2 test with R × C table.Results KS lesions in patients with classic KS were confined to the skin, without mucosal, lymph node or visceral involvement.Lesions in AIDS-KS patients were not only confined to the skin and superficial lymph nodes, but also oral mucosa involved in 12 cases and internal organs involved in 7 cases.Liver and lung involvement was more common.The CD4+T lymphocyte count in patients with AIDS-KS was (200.8±166)/μL.All 15 AIDS cases with CD4+ T cell count less than 200/μL developed opportunistic infections.CD4+ T lymphocyte count of patients with classic KS was (562.52±222.66)/μL and the 16 patients with CD4+T lymphocyte count greater than 350/μL had no opportunistic infections.The results of immunohistochemistry showed that the positive expression rate of caspase-3 protein in KS tissues in patients with AIDS-KS was 68.2%, in patients with classic KS was 100.0%, with significant difference between two groups (χ2=7.37, P=0.01).The positive expression rate of E-cad protein in KS tissues in patients with AIDS-KS was 72.7%, in patients with classic KS was 100.0%, with significant difference between two groups (χ2=5.18, P=0.03).Western blotting showed that the gray value of caspase-3 in the KS tissue of patients with AIDS-KS was 0.55±0.36, and that in patients with classic KS was 0.86±0.56, with significant difference between two groups (t=-2.070, P<0.05).The gray value of E-cad in the KS tissue of patients with AIDS-KS was 0.54±0.41, and that in patients with classic KS was 0.85±0.45, with significant difference between two groups (t=-2.060,P<0.05).Conclusions There are differences in the protein expressions of caspase-3 and E-cad in tumor tissues of patients with AIDS-KS and classical KS in Xinjiang Uygur patients with Kaposi's sarcoma, which may correlate with a faster progression and a higher mortality rate for AIDS-KS.

Objective To analyze the clinical features of AIDS-related Kaposi's sarcoma (AIDSKS) patients in Xinjiang uygur autonomous region,and to evaluate the efficacy of highly active antiretroviral therapy (HAART) and systemic chemotherapy.Methods Clinical data on 70 AIDS-KS patients admitted in Sixth People' s Hospital of Xinjiang Uygur Autonomous Region from January 2008 to June 2013 were reviewed.The demographic characteristics,progress of KS,extent of lesions,treatment and prognosis of these patients were retrospectively studied.The efficacy of different therapies and the prognosis were compared using t or x2 test.Results The average age of 70 AIDS-KS patients was (40 ± 10) years,and the ratio of male and female was 2.5 ∶ 1.Sixty-six out of 70 (94.29％) patients were Uyghur ethnic.Among 70 patients,23 cases (32.9％) were rapidly progressed.Lesions often progress to plaques and nodules affecting head,neck,trunk,limbs and internal organs.Among 64 evaluable cases,the fatality rate in treatment group was 7.7 (4/52),which was significantly lower than that in patients who did not receive treatment (4/12,x2 =5.861,P ＜ 0.05).After 6-month of HAART or HAART sequenced by chemotherapy,CD4+ T lymphocyte counts were significantly increased (t =35.258 and 42.528,P ＜0.01),while it was decreased or remained in low level in patients received other treatments or untreated.HAART sequenced by chemotherapy,chemotherapy sequenced by HAART,and chemotherapy alone were effective for all AIDS-KS patients,while the effective rate in patients received HAART alone was 86.4％ (19/22),and immune reconstitution inflammatory syndrome (IRIS) was appeared in 45.5％ (10/22)patients.Conclusions AIDS-KS more frequently occurs in young Uighur males.The lesion may affect various organs,and some progress rapidly.HAART plus chemotherapy may reduce the incidence of IRIS and improve the prognosis.

Objective To investigate the difference of expression and distribution of Toll-like receptor 4 (TLR4)between acquired immunodeficiency syndrome (AIDS)-related Kaposi sarcoma (AIDS-KS)and classic Kaposi sarcoma (KS).Methods The subjects were collected from The First Teaching Hospital of Xinjiang Medical University from January 2010 to January 2012,including 11 patients with AIDS-KS,7 patients with classic KS,7 human immunodeficiency virus (HIV)infective patients without KS and 7 healthy volunteers.Clinical data and biopsy specimens were collected.The expression of TLR4 was detected by immunohistochemical method.Statistical analysis of mean between two groups was conducted by using t test.Statistical analysis of ranked data was conducted by using U test.Comparison between groups was analyzed with one-way ANOVA.Results The expressions of TLR4 were both strongly positive in patients with AIDS-KS and classic KS,and mainly in alien vascular endothelial cells (VEC)and tumor cells.TLR4 was located on cell membrane and cytoplasm,and also in the nucleus. TLR4 was rarely expressed in the skin tissue in HIV infective patients without KS and healthy volunteers. The expressions of TLR4 in AIDS-KS,classic KS,HIV infective patients and healthy volunteers were 0.242±0.443,0.208±0.059,0.072±0.042 and 0.065 ±0.031 ,respectively (F =33.70,P <0.01 ). The expressions of TLR4 were lower in HIV infective (AIDS ) patients without KS and healthy volunteers.The expression level of TLR4 was significantly higher in KS patients compared with non-KS patients (t=4.097,P =0.003).However,TLR4 expression had no significant difference between AIDS-KS and classic KS patients (t =0.480,P =0.638).The expression of TLR4 was significantly higher in AIDS-KS patients compared with HIV infective patients (t=2.900,P =0.010).Conclusions The study identifies that TLR4 is highly expressed in patients with AIDS-KS or classic KS.It is suggested that TLR4 may play an important role in occurrence and development of KS.