Chronic kidney disease(CKD)is increasingly recognized as a global public health problem.Uncontrolled complications of CKD,especially cardiovascular diseases,contribute greatly to the premature death and unfavorable prognosis.Recent evidence shows that CKD complications may occur earlier than previously thought.CKD complications deserve early detection and active treatment.Periodical follow-up and regular check should be done to adjust the therapeutic condition.Clinical practice guideline or recommendation based on evidence-based medicine is essential for management of CKD complications.Personalized treatment should be considered to improve survival and quality of life,and to make patient return to society.

To determine whether increased expressions of gelatinase A(MMP-2) and gelatinase B(MMP-9) occur in vivo in autoimmune MRL/lpr mice model and to investigate the modulation effects of "shenle." Shenle (4g?kg -1 ?d -1 ,orally) or methylprednisolone(MPS,25mg?kg -1 ?d -1 ,ip)was administered daily to MRL/lpr mice at the age of 8 weeks. The activities of MMP-2/9 by gelatin zymography were compared in kidney protein extracts and urine. After treatment for 20 weeks, a progressive reduction in positive proteinuria number/total mice (40% vs.33 3% vs.80%, proteinuria over 300 mg/dl as positive) and an elevated survival rate (70% vs.80% vs. 50%) were found in "shenle" and MPS groups compared with the control group. Histological analysis of kidney tissues indicated that both "shenle" and MPS could inhibit the mesangial proliferation and renal sclerosis. Using SDS-PAGE gelatin zymography, we have identified increased expressions of both latent and activated form enzymes of MMP-2/9 in urine and kidney extraction. Immunohistochemical staining showed both MMP-2 and MMP-9 were obviously up-regulated within glomerulus in control group. "Shenle" as well as MPS suppressed the expression of both latent and activated form of MMP-2/9. These in vivo results suggested that MMP-2/9 expressions might play an important role in murine lupus nephritis. "Shenle" delayed the development of glomerulonephritis and improved survival in MRL/lpr mice probably by suppressing the expressions and activities of MMP-2/9.

To determine thrombin activation and fibrin deposition in the development of lupus nephritis in MRL lpr/lpr mice and the inhibitory effects of "shenle". "Shenle" (4g/(kg?d) orally) was administered daily to MRL lpr/lpr mice at the age of 8 weeks. After treatment for 20 weeks, we compared thrombin receptor (Protease Activated Receptor-1, PAR-1) expression with immunohistochemistry and fibrin deposition with MSB(Martius-Scarlet-Blue)staining in renal sections. PAR-1 mRNA expression was analyzed with RT-PCR method in the two groups. With the development of murine lupus nephritis, we observed an increase in thrombin receptor mRNA and severe fibrin deposition in renal tissue in the control group, while thrombin receptor protein expression was strikingly downregulated, suggesting its continuous activation and degradation. "Shenle" inhibited PAR-1 activation significantly and it was correlated with reduced fibrin deposition. These results suggested that thrombin activation may play an important role in the development of glomerulonephritis in MRL-lpr mice. "Shenle" ameliorated the murine renal lesions probably by inhibiting thrombin receptor activation and fibrin deposition.

Objective To determine the changes in expressions of thrombin receptor and fibrin deposit in glomeruli during the process of senility. Method Rats were divided into 3 groups (8 rats in each group): 3-month-old group (3m), 12-month-old group (12m) and 24-month-old group (24m). Fibrin deposition was detected by Martius-Scarlet-Blue staining and direct immunofluorecence method. Immunohistochemical studies were performed to detect the expression of thrombin receptor (PAR-1) and transforming growth factor-? (TGF-?). Semi-quantitative PCR was performed to detect the changes in PAR-1 mRNA expression. A quantitative analysis of the expressions was performed by image analysis system. Result Significant pathological changes were found in glomeruli during the process of senility. Fibrin deposition was not observed in glomeruli in different groups. Significant expression of PAR-1 was found in glomerular endothelial cells, mesangial cells and epithelial cells in 3m rats. On the contrary, in 24m rats, PAR-1 expression in glomeruli was significantly decreased. Expression of TGF-? was increased with senility in glomeruli. PAR-1 gene expression, barely detectable in control tissue, was strikingly increased in 24m rats. Conclusion Thrombin receptor activation could be found in glomeruli of senile rat, and it is independent of fibrin deposition. Activation of PAR-1 may play an important role in the process of renal senility.

Objective To investigate the effects of the expression of integerin-linked kinase (ILK) on connexin 43 (Cx 43) in rat mesangial cells (RMCs). Methods RMCs were divided into three different groups (6 for each group): RMC group, ILK-con siRNA group and ILK-siRNA group. ILK siRNA was synthesized, and then transfected into RMCs by LipofectAMIN 2000. RMCs were transiently transfected with ILK-con siRNA, ILK-siRNA and lysed 24h later, and mRNA was then extracted and detected by reverse transcription-polymerase chain reaction (RT-PCR) analysis using ILK and Cx43-specific primers, and an aliquot of protein from each sample was subjected to western blot analysis using ILK and Cx43 antibodies. Cells were seeded into 96-well plates (2?103 cells/well) and then transfected with ILK-con siRNA and ILK-siRNA. After incubation for 24, 48 and 72 hours, respectively, 20?l of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (5mg/ml) was added into each well and incubated for 4 hours. Subsequently, 150?l of dimethyl sulfoxide (DMSO) was added to each well to dissolve the formazan crystals, and the absorption at 492nm was measured. Results ILK mRNA and protein levels decreased by 30%-50% after being transiently transfected with ILK-siRNA, while Cx 43 mRNA and protein levels increased by 30%-40% and 60%-70%, respectively. The viabilities in ILK-siRNA infected RMCs at 24, 48 and 72 hours were significantly higher than that in ILK-con siRNA infected RMCs. Conclusion Inhibition of ILK pathway will up-regulate the expression of Cx 43 and the viability of RMCs, implying that the regulation of connexin 43 might possibly be achieved via ILK pathway.

Objective: Full expression of class I HLA molecules on tumor cells is pivotal in priming the tumor antigen-specific CTLs for effective iramunotherapies. Tumor cells display abnonnal expression of HLA class I molecules in human ovarian carcinoma. Methods: In this study, the expression of class I molecules in human ovarian cancer cells was determined by using Western bloting, immunohistochemistry testing and flow cytometry. The mRNA of TAP (transporter associated with antigen processing) and LMP(low molecular weight polypeptide) were exannined at the same time by RT-PCR. Results: It has been shown that class I molecules were unable to be, or only weakly, expressed in five of eight ovarian cancer cell lines. On these cell lines abnormal in class I expression, no or only little mRNA were detected for TAP1, TAP2, LMP2 and(or) LMP7 genes. Class I expression, however, could be partially recovered by incubating the tumor cells at 25℃ when compared with those at 37℃ . Conclusion: The results suggested that the dysfunction of TAP and IMP genes, following by a defective expression of class I molecules, might be one of the mechanisms enable ovarian cancer cells to escape from immune surveillance.

BACKGROUND:Co-culture withembryonic stem cels or embryonic tissues can induce differentiation of carcinoma cels into normal epithelial cels or decreasemalignancyof carcinoma cels.Acelular embryoid bodies retain the structure and important cytokines of embryonic tissues.
OBJECTIVE:To prepare acelular embryoid bodies from mouse embryonic stem cels and to investigate their effects on differentiation of mouse Lewis lung carcinoma cels at three-dimensional culturein vitro.
METHODS:Mouse embryonic stem cels(D3)were dynamicaly cultured for 7 days to produce embryoid bodiesfolowedbydecelularization with 0.1% sodium dodecyl sulfate. Mouse Lewis lung carcinoma cels were co-cultured with acelular embryoid bodiesas test group or culturedinthree-dimensionalmatrigel mediumfor 7 days as control group, respectively. Cel proliferation and expression of E-cadherin were detected by immunohistochemical staining and western blot assay, respectively. In addition, mRNA expressions ofSlug and E-cadherin were observed using RT-PCR technology.
RESULTSAND CONCLUSION:Uniform mouse embryoid bodieswere successfuly prepared, andwere completely decelularized with sodium dodecyl sulfate. After 7-day three-dimensionalmatrigelculture, in the control group,multicelular tumor spheroidswere formed,accompanied byahigherKi67positive rate;Lewis lung carcinoma cels in the test group were repopulated in the acelular embryoid bodies showing significantly lowerKi67positive rate. Compared with the control group, the absorbance ofPaxilin in the test group was significantly smaler, and the absorbance of E-cadherin was significantly higher (P< 0.05). Besides, mRNA expressions of Slug and E-cadherin were significantly decreased and increasedin the test group compared with the control group, respectively(P< 0.05). These findings indicate that the acelular embryoid bodies can promote differentiation of mouse Lewis lung carcinoma celsinthree-dimensional culturein vitro.

Objective To describe the clinical application of microvascular anastomotic device in head and neck reconstruction.Methods From July,2013 to November,2013,microvascular free flaps were transferred to reconstruct the defects simultaneously after tumor resection of head and neck region in 12 cases in Department of Oral and Maxillofacial Surgery,Peking University School of Stomatology.Microvascular anastomotic coupling devices (MACD) were used in vascular anastomosis.The clinical data were collected and analyzed,including the selection of free flap,diameter of donor and recipient vessels,type of MACD,time of anastomosis,instant patency of anastomosis.The flap was monitored closely after operation and the final survival rate was calculated.Results Twelve microvascular free flaps were done in this series,including 6 fibula flaps,4 forearm flaps and 2 anterolateral thigh flaps.Totally 17 MACD were used by end-to-end anastomosis in this series,including 5 arterial anastomosis and 12 venous anastomosis.The anastomose time using MACD was from 4 to 10 minutes,with a median time 6.8 minutes.The instant patency rate of anastomosis was 100％.There were some blood leakages near the anastomotic stoma in 1 arterial anastomosis using MACD.It was resolved successfully by changing a new MACD.Conclusion Our primary clinical experience showed that the MACD was well suited to the microvascular reconstruction of head and neck defect.The feasibility and reliability was confirmed by our clinical cases.It should be recommended as a safe,fast and reliable adjuvant anastomotic instrument for free tissue transfer.

Objective To investigate the effect of different concentrations of rapamycin on the proliferation and apoptosis of glomerular mesangial cells(GMCs)and to investigate the mechanism. Methods GMCs were treated with different concentrations of rapamycin(1 μg/L,2 μg/L,4 μg/L,8 μg/L,16 μg/L).After treatment for 24 h,48 h and 72 h,cell proliferation was assessed bv MTT colorimetric assay and the growth curve was traced.After treatment for 72 h,the cell cycle distribution and the apoptotic rate of GMCs in different concentrations of rapamycin were analyzed bv flow cytometry.The effects of different concentrations of rapamycin on the mRNA and protein expression of p27 and p53 were detected by RT-PCR and Western blot respectivelyResult The low dose of rapamycin(1 μ/L)could signiticanfly inhibit the proliferation of GMCs and showed no effect on apoptosis.The high dose of rapamycin (8-16 μg/L)could significantly increase the apoptotic rate of GMCs.Rapamycin could increase the mRNA and protein expression of p27 and p53. Conclusion Rapamycin can inhibit GMCs proliferation and promote GMCs apoptosis by increasing the expression of p27 and p53.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven eases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi- quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpeint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperurieacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial- tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average foUow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16～89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.