Objective: To investigate the effects of simvastatin on K562 cells apoptosis and elucidate its molecular mechanisms. Methods: K562 cells were treated with various concentrations of simvastatin for different time,and then AnnexinⅤ-FITC/PI staining was performed to confirm the apoptosis of K562 cells. Flow cytometry was employed to measure ROS level;mutant P53 protein expression level in K562 cells was measured by immunohistochemistry method. RT-PCR was employed to analyze the mRNA expression levels of c-jun. Results: K562 cells were induced to undergo apoptosis after 5,10 ?mol/L and 20 ?mol/L simvastatin treatment for 48 h and 72 h. After the treatment of 10 ?mol/L and 20 ?mol/L simvastatin for 48 h,compared with the control group,ROS level in K562 cells increased obviously. Immunohistochemistry results indicated that simvastatin could inhibit protein expression levels of mutant-type P53. RT-PCR results showed that simvastatin could increase the mRNA expression levels of c-jun significantly. Conclusion: Simvastatin induces K562 cells apoptosis by down-regulating mutant P53 protein and up-regulating c-jun mRNA expression which might be caused by changes of intracellular redox state.

Objective To evaluate the effect of various sampling errors on ELISA test results. Methods Standard sample volume,standard sample volume reducing 1,2,3,4 μL or adding 1, 2,3 μL were respectively pipetted into the wells of a microplate,follwed by routine operation of ELISA test. Then the influence of various sampling errors was analyzed on ELISA test results of HBsAg, HCV and TP. Results S/CO value was increased with the increase of sample volume. The statistical difference of mean S/CO value of HBsAg and TP was only found between sample volume adding 3 μL group and control group(P<0.05). For HCV result, there were significant differences between standard sample volume adding 2,3μL or reducing 3,4μL groups and control group(P<0.05), while no obvious differences were found in the other groups(P>0.05). The difference of mean positive rate between ex-perimental groups and control group showed an increasing tendency with the reduction of sample vol-ume,and significant differences in HBsAg, HCV and TP results were also found between sample vol-ume increase groups and reduction groups(P<0.05). Conclusion Various sampling errors influence ELISA test results to different degrees,and the extent increases with the reduction of standard sample volume.

Objective To study the clinical characteristics of acute leukemia(AL) patients with cross‐lineage antigen expression . Methods Patients were diagnosed and classified by morphology ,cytochemistry and immunology assay ,and prognostic acting factor were also analyzed .Results According to FAB standards ,acute myeloid leukemia(AML)‐M2 ,acute lymphocytic leukemia (ALL)‐L2 and no‐classified type were common in 320 patients with cross‐lineage antigen expression .The immunophenotype with B and my‐eloid mixed expression was the most common(176 cases) ,followed by cross expression of antigen T and myeloid(131 cases) ,and the co‐expression of B ,T and myeloid antigen was found in only 10 cases .In lymphoid antigenpositive AML(Ly+ AML) ,CD19 anti‐gen was the most common among B lineage ,CD7 was the most common in T lineage .In myeloid antigen positive ALL(My+ ALL) , CD33 was the most common myeloid antigen .Forty‐five cases were with mixed expression of myeloid antigen and CD56 expression . Correlation existed between CD7 and CD34(P< 0 .05) ,CD19 and CD34(P< 0 .05) .There were 9 patients with CD34 ,CD7 and CD19 co‐expression ,7 patients with CD34 ,CD7 and CD56 co‐express .In Ly+ AML patients ,23 cases were with recurrent chromo‐some abnormality ,including 11 cases with t(8 ;21)(q22 ;q22) ,RUNX1‐RUNX1T1 ,3 cases with t(15;17)(q22 ;q11‐12) ,PML/RAR ,6 cases with bone marrow eosinophilia inv(16)(p13 ;q22) ,CBF beta /MYH11 ,and 3 cases with t(9;11)(p22;q23) , MLLT3‐MLL .In My+ ALL ,15 patients were with recurrent chromosome abnormality ,including 9 cases of B‐ALL with t(9;22) (q34 ;q11 .2) ,BCR‐ABL1 ,3 cases of B‐ALL with t(v ;11q23) ,MLL rearrangement ,and 3 cases of T‐ALL with 14q11 .2 .Among the presence of reproducible chromosomal abnormalities in AL ,the antigen expression of mistranslation was still with a certain fea‐ture:Ly+ AML patients often mistranslated CD19 ,CD56 ,CD2 ,and My+ ALL patients often mistranslated CD13 and CD33 .Com‐pared with the lymphoid antigennegative AML(Ly - ALL) group ,CD7+ AML group ,CD19+ AML group and CD56+ AML group had significant difference in survival curves(with P value of 0 .01 ,0 .02 and 0 .02) .There was no significant difference in survival curves between myeloid antigen negative ALL(My -ALL) group and CD13/33+ ALL group(P<0 .05) .CD7 was also positive com‐monly(53 cases) and related with CD34(P<0 .05) .So it significantly influenced the prognosis .If patients were with co‐expression of CD34 ,CD7 and CD19 ,the prognosis could be worse .Conclusion AL with cross‐lineage antigen expression might be a special type and confirmed by immunotype .Furthermore ,expression types of differentiation antigen could be critical for prognosis and sur‐vival .

Objective To know the present sanitary status of bathing beach in Haikou,and provide the scientific data for improving the assessment of microorganisms contamination of bathing beach. Methods The routine water quality data and related meteorological data of the bathing beach were collected from 2007 to 2009. In July 8th-10th 2009,a high grade bathing beach and an ordinary one were selected,four sampling sites were selected where the contamination by feces were often seen,the number of swimmers was the largest,the samples were collected from the deep-water areas and sand beach respectively,and fecal coliform and enterococcus were examined. Results The concentration of fecal coliform was(1 085.8?538.8)cfu/L in water and(120 000.0?32 659.9)cfu/1 000 g in sand,and the E.faecalis was(32.5?19.1)cfu/L in water and(2 425.0?689.8)cfu/1 000 g in sand. The numbers of the two kinds of bacteria in water decreased with the increase of distance from the land. The concentration of bacteria was positively correlated to the day's rainfall(P

Objective To investigate the clinical value of serum iron(Fe),total iron binding capacity(TIBC),serum ferritin(SF), folic acid(FA)and vitamin 12(ViB12 )in the diagnosis and treatment of chronic renal failure(CRF).Methods Fasting blood sam-ples were collected from 72 patients with CRF and 83 normal controls.Then the serum SF,ViB12 and FA contents were measured by the ARCHITECT i2000SR fully automatic chemiluminescence immunoassay analyzer;serum Fe and TIBC were detected by the VITORS FS 5.1 dry biochemical analyzer;RBC,HGB,HCT and MCV were analyzed by the Mindray BC-6800 complete automated blood counter.The detection results were performed the statistical analysis by the SPSS16.0 software.Results The levels of TIBC,RBC,HGB and HCT in the CRF patients were significantly lower than those in the control group(P <0.05);while the SF and ViB12 levels were significantly higher than those in the control group;serum Fe and MCV had no statistical difference compared with the control group(P >0.05);the FA level in the female patients was lower than that in the control group,but which in the male patients had no statistical differences compared with the control group(P >0.05).Conclusion The indexes of anemia associat-ed metabolin in the CRF patients can provide certain reference value for the diagnosis and treatment of chronic renal fail and has cer-tain guidance significance for correcting anemia caused by renal insufficiency.

Objective To investigate the correlation between infertility and TORCH infections ,analyse the possible influence factors .Methods TORCH infections of 2343 cases of pregnant women and 1356 cases of infertility women were detected by chemi‐luminescence method ,the positive rates of TORCH‐IgM ,IgG were compared .Results RV ,HSV infections of infertility women were higher ,mainly in 30 -34 years group .TORCH infections of infertility women among seasons were of significant difference (P<0 .05) .TORCH infections of infertility women have correlations with schooling levels ,source of family ,and history of expo‐sure to animals (P<0 .05) .Conclusion The less education ,rural aera ,and history of exposure to animals were high risk factors of TORCH infections ,so screening for TORCH infections of infertility women is very necessary .

Objective To evaluate the methods of measuring platelet-associated antibody PAIgG/ PAIgA/ PAIgM by flow cytometry(FCM) and enzyme linked immunosorbent assay(ELISA), and to investigate their diagnostic value for patients with idiopathic thrombocytopenic purpura(ITP).Methods With FCM and ELISA respectively, PAIg on the platelet membrane and in plasma were measured in 19 patients with ITP and 17 healthy volunteers, and were compared with each other in order to find out whether there were differences in these groups.Results FCM and ELISA measurement in patients with ITP were significantly higher than those in control group (P0.05). Compared with the results of ELISA, the positive percentage of PAIgG measured by FCM(84%) in ITP patients was slightly higher than that by ELISA(79%).Conclusion The platelet-associated antibodies of PAIgG/ PAIgA/ PAIgM, especially PAIgG,are important for diagnosing ITP. FCM, in combination with ELISA, may improve the reliability and the positive percentage of detection in ITP patients.

Objective To use fluorescent quantitative polymerase chain reaction (FQ-PCR) todetect the level of cyclin D1 gene (CCND1), and to investigate its clinical application in the diagnosisand monitoring of breast cancer. Methods FQ-PCR was applied to detecting the expression level ofCCND1 gene in peripheral blood from 15 healthy females, 30 patients with benign breast disease and81 patients with breast cancer with β2-microglobin as the internal control. Results There were no sig-nificant difference in the level of CCND1 expression between healthy control group and benign breastdisease group (P>0. 05). The level of CCND1 expression was significantly higher in breast cancergroup than that in healthy control group and benign breast disease group (P<0.05). The difference inβ2-microglobin was statistically insignificant among three groups (P> 0. 05). The positive rate was45.7% in breast cancer group, and 0 in benign breast disease group. Conclusion FQ-PCR is a rapid,sensitive and specific method for quantitative determination of CCND1 gene, which may he used as anavailable tool for the diagnosis, monitoring of therapeutic effect and metastasis as well as prognosis e-valuation of breast cancer.

Objective To interpret the clinicopathological features and the key factors for diagnosis of polyomavirus-associated nephropathy (PVAN).Methods Clinicopathological data of 13casesof polyomavirus-associatednephropathyduring2008-2011inour hospitalwere retrospectively analyzed.Three cases received repeat biopsy.The clinicopathological features were analyzed according to thelight microscopicsceneandSV40-Timmunochemicalexpression.Results Recipients had a peak incidence of PVAN in 12 to 18 months period after renal transplantation,accompanied by elevated serum creatinine.Due to the progression of the disease,3patterns of histological findings could be identified.The early lesion was confined to the collected ducts,with slightly inflammatory infiltration in medullary interstitium,viral inclusions were not necessarily seen.The only findings could be enlarged nuclear and irregular arrangement of the tubular epithelial cells.At the developing stage,prominent tubulointerstitial nephritis was detected,and the involved tubules extended to other segments of renal tubule,even the parietal epithelial cells of Bowman's capsule could be compromised.The epithelial cells shed off,leading the tubular basement membrane exposed.Typical intra-nuclear inclusions as well as variable nuclear changes were found.At the end stage,the allograft showed notable chronic tubulointersititial change,with diffuse tubular atrophy and interstitial fibrosis.Although in this period,typical viral inclusions were rare, stillIHCshowedpositiveexpression of SV40-T. After immunosuppressantreductionor exchange,2 cases developed renal failure,4 cases showed sustained increment in serum creatinine,while 7 cases had a stabilized serum creatinine level.Conclusions Polyomavirus-associated nephropathy can display uneven pathological changes,as well as the morphology of the infected epithelial cells.Segments of the involved tubule are associated with the course of disease.Reduction of immunosuppressant at the early stage has a favorable effect.A prompt renal biopsy should be done in renal transplant recipient if who shows increased serum creatinine,and a routine polyomavirus immunohistochemical staining should be applied as well.

Objective To analyze the apoptosis mechanisms of K562 cells in a PI3K-AKT-dependent manner.Methods K562 cells were cultured in vitro for experiments below:the proliferation assay of K562 cells detected by MTT,the analysis of apoptosis rate and cell cycle of K562 cells measured by flow cytometry(FCM),the construction of chronic myeloid leukemia(CML) animal model through the subcutaneous inoculation of K562 cells to 12 BALB/c-nu/nu nude mice,the early apoptosis changes of K562 cells detected by TdT-mediated dUTP nick end labeling(TUNEL) and the in vitro and in vivo changes of N-ras,PI3K,AKT1,IKK-?,NF-?B1 at transcription level detected by RT-PCR.Results Simvastatin inhibited the proliferation of K562 cells and induced their G0/G1 arrest and significant apoptosis.N-ras and most genes of PI3K-AKT signal pathway were expressed differentially in vitro.K562 cells on nude mice could be induced to apoptosis by simvastatin and the apoptotic index increased with the dose accumulation of simvastatin(P=0.000).The differential mRNA expression changes of N-Ras and most genes of PI3K-AKT signal pathway in K562 cells were observed after treatment of simvastatin at different doses(P=0.000 or P=0.003).However,mRNA expres-sion of the genes in PI3K-AKT pathway in vitro differed to that in vivo.Conclusion The differential expression at transcription levels of N-ras and the most genes in PI3K-AKT pathway that are involved in anti-apoptosis in K562 cells,to some degree,indicates that simvastatin can induce the apoptosis of K562 cells in a PI3K-AKT pathway-dependent fashion in vitro.The animal experiments confirm that there are different mechanisms of simvastatin in inducing the apoptosis of K562 cells in vitro and in vivo.