Chronic kidney disease ( CKD ) is a worldwide public health problem.Glomerular filtration rate ( GFR ) is a key indicator for early diagnosis and accurate classification of CKD , how to estimate GFR accurately and conveniently has been a difficulty and hot-points.We describes the development of several major eGFR equations based on creatinine and cystatin C , analysis the impact of the different research methods on performance of equation , and discusses the issue in research and application of eGFR in China.Accordingly make the following recommendations ( 1 ) Expand the study size ( multiple centers) and participants number , then develop and validate eGFR equations based on Chinese population ;(2) Standardize the gold standardof GFR in the study and unify the analysis and evaluation methods;(3) Promote the consistency and standardization of creatinine and cystatin C which is the basis for the wide range of applications of the eGFR formula.

AIM: IL-12 acts upon T lymphocytes and activates its receptor complexes of ?1/?2 ,and so IL-12 can regulate TH1/TH2 balance. Our study is aimed at IL-12-inducing apoptosis of T cells and expression and signal conduction of Bcl-2 during T cell apoptosis. METHODS: The apoptosis of T cells was detected by Annexin V staining cytometry and the expression of Bcl-2 under different inhibitors were detected by the method of semi-quantitative PCR. RESULTS: IL-12 can induce the human leukemic T cell line(TIB-152) and the human lymphoma T cell line(HTB-176) and the normal human T cells to undergo apoptosis. The Bcl-2 expression at 6 hours of treatment with IL-12 increased aparently,and reached the max at 24 hours. But IL-12 did influence Bcl-2 expression. IL-12 can induce T cells to undergo apoptosis which is characterized by early membrane changes. CONCLUSION: The inducing effect is correlated with the concentration of IL-12 and the maturation of T cells. Bcl-2 takes part in the progression of T cells' apoptosis as a apoptosis mediator.

AIM: To illustrate the expression of hypermethylation p15 gene in 53 non-Hodgkin’s lymphoma (NHL). METHODS: The methylation of p15 gene in 53 cases of non-Hodgkin’s lymphoma was detected by using methylation-specific PCR (MSP) technique. RESULTS: 18.9% (10/53) in NHL were methylation in p15 gene. p15 gene was frequently in high malignant NHL patients (27.3%) compared with in low malignant patients (0%). CONCLUSION: The result suggested that the hypermethylation of p15 may play an important role in non-Hodgkin’s lymphoma.

AIM: To investigate the variation and distribution of abnormaly methylated p15 INK4B in myelodysplastic syndrome (MDS) and its subgroups. METHODS: The abnormal methylation of p15 INK4B in 32 cases with MDS was studied using methylation-specific PCR (MSP). RESULTS: The positive rate of abnormal methylation of p15 INK4B was about 50% in MDS. The ratios in subtypes were: 0% (0/6) in RA,20% (1/5) in RA-S,57.1% (4/7) in RAEB,74.1% (5/7) in CMML,85.7% (6/7) in RAEB-t, respectively.It was worth noticing that 4 cases represented abnormal methylation of p15INK4B during their transformation and progression into AML. CONCLUSION: The abnormal methylation in p15 INK4B might be one of the causes of MDS, which was related to pathologial process of MDS.Every subtype was not solitary classification completely. Abnormal methylation of p15 INK4B was apt to occur accompanying the progression and transformation of the subtypes.

AIM: To study the effect of IL-12 on T lymphocytes apoptosis, the expression of Fas/FasL and TNFR/TNF?. METHODS: Terminal dUTP nick end labeling(TUNEL) and Annexin V assay were used. Anti-TNFR were labeled with FITC, anti-CD95 was labeled with PE and Anti-FasL with biotin. Three kinds of T cells (HTB176,TIB152 and human normal T cells) were analysed through flow cytometry. RESULTS: At 1st hour after being treated with IL-12, the expression of FasL protein and FasL mRNA in HTB176 and TIB152 began to increase and reached peak value in 24 hours. In the normal T cells, FasL just began to increase in 1 hour and maintained stability in 6, 12 and 24 hours through the later experiment period. All three kinds of T cells displayed no change in the expression of CD95 and TNFR/TNF? under the stimulation of IL-12. CONCLUSION: Expression of such apoptosis regulating factors were different in the apoptosis of T cells induced by IL-12.

ted by the ECI analyzer.

Objective To validate the analytical performance of three Cys C reagents with particle-enhanced turbidimetric immunoassay(PETIA) method used on the automatic biochemistry analyzer for preliminary clinical application.Methods The performance of three Cys C reagents (labeled as A, B, C) with PETIA method from Shanghai Jing Yuan Co., Beijing Leadman Co. and Beijing Jiuqiang Co. on OlympusAU2700 automatic biochemistry analyzer were assessed.According to the standard of CLSI EP6-A, EP15-A and EP7-P, the precision, linearity range, disturbance (bilirubin, hemoglobin, chyle) were assessed, and compared with those of Cys C reagent based on particle-enhanced nephelometric immunoassay(PENIA) from Dade Behring Co.. The reference ranges for Cys C in serum of 120 healthy individual were evaluated.Results The within-run CVs of the three reagents (A, B and C) were 3.08%-3.2%, 2.3%-4.15% and 1.38%-1.53% respectively.The total CV in A, B and C were 3.29%-3.44%, 2.65%-5.18% and 1.67%-1.69% respectively, lower than the stated.Limits of quantitative determination (LOQ) of the three reagents were 0.41, 0.23 and 0.07 mg/L, basically meeting the testing requirement.The linearity range was 0.22-7.26 mg/L(r=0.996), 0.20-7.72 mg/L(r=0.999)and 0.20-7.62 mg/L(r=0.997)in the three reagents, which demonstrated a sound linear correlation. For interference tests, no remarkable interference (<±10%) of reagent C was detected when bilirubin≤684 μmol/L, hemoglobin≤9.7 g/L and Chyle turbidity≤6 200 FTU; and no significant interference of reagent B was found when bilirubin≤684 μmol/L, hemoglobin≤6.79 g/L and Chyle turbidity≤6 200 FTU; when bilirubin≤684 μmol/L, hemoglobin≤4.85 g/L and Chyle turbidity≤1 240 FTU reagent A was not interfered significantly. The comparison afte and before the high-speed centrifugation reveals that the average percentage of bias for reagents A, B, C measured Cys C in chylous serum samples of patients was -8.31%, 1.52%, 1.32%, respectively.In method comparison tests, the regression equations of the three reagents compared with Dade Behring PENIA Cys C reagent were as follows:Y=0.787X+0.492 (R2=0.976), Y=1.098X+0.137 (R2=0.982) and Y=1.037X+0.249 (R2=0.996), respectively. Agreement rates of the high Cys C in reagent A, B, C and Dade Behring Cys C reagent were 80% (Kappa=0.615,P=0.000), 100% (Kappa=1.000,P=0.000), 91.2% (Kappa=0.824,P=0.000); While for reference range of preliminary clinical assessment, diagnosis coincidence rate of reagent A increased to 98.8% (Kappa=0.974,P=0.000). Conclusions When used in automatic biochemical analyzer, the three Cys C reagent with PETIA showed high precision,sensitivity, and sound correlation with Dade Behring PENIA reagents.The three reagents are all able to meet clinical test requirements, nevertheless, anti-interference capability were diffierent and the reference range should be further validated.

Objective To investigate the applicability of Cys C-based formulas for prediction of GFR in Chinese patients with CKD.Methods A total of 176 adult patients with CKD including 90 males and 86 females collected from 4 hospitals located in different geographic regions of China (Beijing,Shanghai,Dalian and Changsha) were enrolled in this study from September 2007 to July 2009.The rGFR was measured using 99mTc-DTPA clearance rate two-sample method.Cystantic C was measured by PETIA and PENIA respectively.The results of eGFR in the Larsson formula,Grubb formula,Hoek formula,Filler formula,Stevens formula and Hojs formula were compared with the rGFR to evaluate the calculation coherence,bias,precision,accuracy and the performance of correct phasing of the formulas.Results The mean 99mTc-DTPA clearance was [40.70 ( 19.09 - 86.49)] ml · min-1 · ( 1.73m2 ) -1.Significant difference was witnessed in the evaluation of GFR estimation formulas calculated by PETIA and PENIA (P ＜0.01).ICC and Spearman correlation analysis revealed a significant correlation between eGFR and rGFR.The ICCs of eGFR and rGFR ranged from 0.874 to 0.938.Compared with rGFR,the 30％ accuracy of all the eight evaluation formulas using PETIA and PENIA method were lower than 60％.The percentages of correct phasing in all the 5 stages of CKD were not ideal.With these formulas,percentages of correct phasing from CKD stage 2 to CKD stage 4 were lower than 65％.The eGFRs were underestimated by formulas evaluated by PENIA in CKD stage 1.All the eGFRs were overestimated remarkably by all equations in CKD stage 5.Conclusions None of the eight Cys C based formulas are ideal for estimation of GFR in Chinese CKD patients,and they can not be applied to Chinese patients directly.For this patient population,further studies will be needed to develop a more accurate Cys C-based GFR estimation formula that includes ethnicity,age and other factors.

Objective To explore the localization of epileptogenic focus and select the appropriate surgical procedures for post-traumatic epilepsy. Methods The clinical data of 21 patients with post-traumatic epilepsy were studied retrospectively. Epileptogenic focus was located by comprehensively analyzing data of electro-neurophysiology, neurological imaging and clinical manifestation. Surgical procedures were performed in all patients, including resection of lesion and peripheral cortex in 12 patients, epileptogenie focus resection plus low power bipolar coagulation in five, anterior temporal iobectomy plus amygdalohippocampectomy in three and corpus callosotomy in one. Results All patients were followed up from 6 months to 3 years, which showed satisfactory outcome in eight patients, marked improvement in six, improvement in five and slight improvement in two. The total effective rate was 90%. Conclusions Surgical procedure is important for intractable post-traumatic epilepsy. The good efficacy depends on precise localization of epileptogenic focus and combined application of various surgical procedures.

Objective To investigate the alterations and phenotypes of dendritic cells, inflamma-tory monocytes and macrophages in immunocompetent mice during Pneumocystis murina ( P.murina) infec-tion for further analysis of the function of these cells during P.murina infection.Methods Wild type male C57BL/6 mice at age 6-8 weeks were randomly divided into two groups including the group with P.murina infection and the group receiving sham surgery.The mice without any intervention were used to set up the blank control group.The loads of P.murina strains in lung tissues of each mouse were quantified by TaqMan real-time fluorescence polymerase chain reaction after the infection.Histopathological examination was per-formed to evaluate the degree of inflammation in lung tissues.The numbers of dendritic cells, inflammatory monocytes and macrophages in lung tissues, peripheral blood and bone marrow samples, and the changes of inflammatory monocytes in spleen tissues were measured by flow cytometry analysis.The expression of major histocompatability complexⅡ(MHCⅡ), CX3C chemokine receptor 1 (CX3CR1) and CC chemokine re-ceptor 2 ( CCR2 ) by dendritic cells, inflammatory monocytes and macrophages in lung tissues during P.murina infection were analyzed by flow cytometry analysis.All of the data were collected one, two, three and four weeks after the corresponding treatments.Results The loads of P.murina strains in P.murina in-fected mice were elevated after two and three weeks infection, but decline at week 4 (P>0.05).Significant pathological changes including the alveolar destruction, inflammatory cell infiltration and thickened alveolar septum in mice with P.murina infection were observed under a microscope at week 3 and week 4.Compared to the sham surgery treatment group, the number of CD11c+CD11b+dendritic cells were increased in lung tissues, but decreased in blood samples during P.murina infection ( P<0.05) .The levels of inflammatory monocytes in blood samples fell at week 3 and then rose at week 4 during P.murina infection (P<0.05). No significant difference with the change of macrophages in mice was observed during P.murina infection ( P>0.05).The CD11c+CD11b+dendritic cells in lung tissues of mice with P.murina infection expressed high levels of MHCⅡand CX3CR1, and low levels of CCR2.The inflammatory monocytes in lung tissues of mice expressed high levels of CCR2, moderate levels of MHCⅡand low levels of CX3CR1 during P.murina in-fection.High levels of CX3CR1 and low levels of MHCⅡ and CCR2 were observed in macrophages from lung tissues of mice with P.murina infection.Conclusion Highly expressed CD11c+CD11b+dendritic cells and MHCⅡwere detected in lung tissues of mice during P.murina infection, indicating that CD11c+CD11b+dendritic cells were involved in the host defense against P.murina infection.