Objective   To analyze the characteristics and prognosis of elderly renal transplant recipients.  Methods   The authors included 130 recipients older than or equal to 60 years at the time of operation in elderly group, and the paired 130 patients receiving contralateral renal transplants from the same donors and younger than 60 years in control group. All the patients received renal transplant in Kidney Disease Center of the First Affiliated Hospital of Zhejiang University from Nov 1994 to Dec 2013.  Results   The average age of the patients of elderly group was (63±3) years old, whereas the patients in control group were (41±10) years old. There was no significant difference in sex, type of dialysis, number of mismatched HLA, level of panel reactive antibodies (PRA), percentage of receiving induction therapy or immunosuppressive regimen between elderly group and control group, except that the patients in old group had a longer duration of dialysis. The patients of elderly group had a lower level of serum creatinine than control at the follow up times from 6 months to 24 months after transplant. The doses of immunosuppressives were lower in elderly group compared with the control group whereas the concentration of tacrolimus or cyclosporine was same. The dose of prednisone in old group was lower compared with control after 6 months post - transplantation. The patients of elderly group had high percentages of pulmonary infection and new-onset diabetes mellitus compared with the control group. Until June 2014, the follow-up rate of all patients was 85.4%; the median follow-up time was 70.4 months in elderly group and 79.9 months in control group. There was no significant differencein mortality rate or graft loss rate between elderly group and control group. Pulmonary infection (HR=2.981, P=0.018), hepatitis C virus infection (HR=5.797, P=0.003) and malignancies (HR=5.228, P=0.005) were correlated with the survival rate of the elderly group.  Conclusions   Elderly renal transplant recipients have a similar prognosis compared with the younger ones. Pulmonary infection, hepatitis C virus infection and malignancy are related risk factors for the survival rate of elderly patients.

Objective    To evaluate the nighttime blood pressure(BP) control status of hypertensive Chinese chronic kidney disease (CKD) patients and related risk factors.  Methods    This cross - sectional study enrolled 337 hypertensive CKD in - patients. The clinical and ambulatory BP monitoring (ABPM) data were retrieved from the electronic database of the hospital. High ambulatory BP were defined as >130/80 mmHg (average 24 - hour BP) and >135/85 mmHg (daytime) />120/70 mmHg (nighttime), respectively. Multivariable analysis was used to evaluate the risk factors for lack of nighttime BP control and circadian rhythm.  Results    There were 38.6% of the whole population had average 24-hour BP controlled. But only 22.8% of them achieved nighttime BP control, which was far less than the 50.7% of daytime BP control (P＜0.01). Even among those patients who achieved average 24 - hour BP control shown by ABPM, there were still 44.6% of them with uncontrolled nighttime BP. Multiple analyses showed urinary protein excretion (OR: 1.151, 95%CI: 1.035-1.279) was independent risk factor for lack of nighttime BP control. About 80% of patients presented with non- dipping BP pattern, among whom 37.3% were presented with reverse-dipper pattern. Lack of nighttime BP control was independent risk factor for lack of normal circadian rhythm (both P＜0.001).  Conclusions    Lack of nighttime BP control was common in hypertensive CKD patients and contributed to the abnormal circadian rhythm. ABPM should be performed more commonly in clinical practice to help nighttime BP control in the future.

Objective   To investigate and analyze the mineral and bone disorder (MBD) in the patients with chronic kidney disease (CKD), reveal the change of related indexes of CKD-MBD.   Methods   A cross-sectional study was carried out in the First Affiliated Hospital of Harbin Medical University. From October 2011 to May 2014, 1318 inpatients and hemodialysis outpatients were enrolled. Parameters related to MBD, including serum phosphorus (P), total calcium (t - Ca), intact parathyroid hormone (iPTH) and alkaline phosphatase (AKP) were analyzed. Last, it was analyzed with multiple regression analysis to related factors of the secondary hyperparathyroidism (SHPT) in patients with CKD.  Results   Serum calcium, phosphorus and iPTH had no obvious abnormalities at the early stages of CKD [GFR＞60 ml•min^-1•(1.73 m²）^-1], and relatively stable before GFR＞30 ml•min^-1•(1.73 m²)^-1. After entering the CKD4 stage, serum phosphorus, iPTH increased sharply and serum calcium decreased obviously along with the decreased glomerular filtration rate (GFR). Serum P, t-Ca and iPTH levels were statistically significant in CKD 1 to 5D patients, respectively, serum P: (1.13±0.20) mmol/L, (1.14±0.22) mmol/L, (1.26±0.23) mmol/L, (1.48±0.34) mmol/L, (2.05±0.61) mmol/L and (2.08±0.58) mmol/L; serum t-Ca (mmol/L) (2.35±0.13) mmol/L, (2.35±0.12) mmol/L, (2.35±0.15) mmol/L, (2.26± 0.18) mmol/L, (2.07±0.29) mmol/L and (2.31±0.26) mmol/L; iPTH: 57.8(45.6, 91.8) ng/L, 54.1(37.8, 74.6) ng/L, 71.6(45.8, 102.2) ng/L, 131.1(81.7, 205.1) ng/L, 277.5(173.6, 395.3) ng/L and 354.9 (194.4, 720.3) ng/L; The stepwise logistic regression analysis showed: hypocalcemia (OR=3.32, P＜0.01) and decreased GFR (OR=5.28, P＜0.01) were independent risk factors of iPTH elevation at stage CKD3～5.  Conclusions   From the beginning of the CKD3 stage, serum t - Ca, P, iPTH level began to be relatively abnormal as renal function declined. Hyperphosphatemia, SHPT has not been improved significantly in CKD5D stage patients even with hemodialysis. The regulation of hemodialysis on serum calcium showed "overcorrecting" phenomenon.

Objective   To investigate the factors correlated to coronary artery calcification (CAC) in maintenance hemodialysis (MHD) patients.  Methods   This study included 132 patients(54 females, 78 males), aged 26-94 years, who were on hemodialysis for 10-204 months(median dialysis duration 51.00 months). The parameters including calcium, phosphorus, parathyroid hormone, total cholesterol, low density lipoprotein, triglycerides, C - reactive protein (CRP), klotho, and so on were assessed. Quantification of CAC was determined by multi-slice spiral computed tomography (MSCT), known as the coronary artery calcification score (CACs).  Results   Ninety-two patients (69.70%) had CAC, with CACs ranging from 0 to 13 450.20. More than 30% patients experienced one even a variety of cardiovascular and cerebrovascular diseases. A positive correlation was observed between the degree of CAC and the incidence of cardiovascular and cerebrovascular diseases. Whereas a positive correlation existed between CACs and age (r=0.347, P=0.000), duration of hemodialysis (r=0.245, P= 0.005), systolic blood pressure (r=0.184, P=0.034), diabetes history (r=0.211, P=0.015), phosphorus (r= 0.262, P=0.002), calcium-phosphorus product (r=0.247, P=0.004); and a negative correlation between CACs and klotho level (r=-0.294, P=0.001). Multivariate logistic regression analysis showed that the main factor influencing the degree of CAC in MHD patients was age.  Conclusions   CAC is common and widespread in hemodialysis patients, who are often accompanied by cardiovascular and cerebrovascular diseases. The prevalence rate of cardiovascular and cerebrovascular diseases increases with the aggravation of CAC degree. Age, duration of hemodialysis, systolic blood pressure, diabetes history, disturbance of calcium and phosphorus metabolism and klotho are correlated with the severity of CAC. Age is an independent risk factor of CAC degree.

Objective   To investigate the effect of pretreatment with U75302, antagonist of leukotriene B4 receptor 1 (BLT1), on cisplatin induced acute kidney injury in mice and its immuno- regulatory mechanism.  Methods   Healthy C57BL/6 mice were randomized into four subgroups: 1. healthy control group; 2.cisplatin group; 3.U75302 control group; 4.cisplatin + U75302 group, n=6. Group 2 and 4 received intraperitoneal injection of cisplatin (20 mg/kg) on day 0, group 3 and 4 received intraperitoneal injection of U75302 (5 μg/mouse) on day 0 and day 2. Mice were sacrificed on the 3^rd day and blood and kidney were collected. Renal function and histological changes were estimated, the infiltration of immune cells were determined by flow cytometry, the level of peroxidase (MPO) in kidney were determined by colorimetry, relative expression of TNF - α, IL - 1β, CXCL1, CXCL2 were detected by Real-time PCR.  Results   Compared with healthy control group, levels of BUN, Scr were higher in cisplatin group with serious tubular structural damage. There were more neutrophils, macrophages, CD4⁺ T lymphocytes, CD8⁺ T lymphocytes in kidneys of cisplatin group, the level of MPO and relative expression of TNF-α, IL-1β, CXCL1, CXCL2 were also higher in cisplatin group. Compared with cisplatin group, lower BUN [(17.75±1.80) mmol/L vs（42.6±6.66）mmol/L, P＜ 0.05], Scr were found in cisplatin + U75302 group with less tubular structural damage. Meanwhile, U75302 reduced infiltration of neutrophils [(146±13)×10³/g vs (296±66) ×10³/g, P＜0.05], macrophages [(245±13)×10³/g vs (420±78)×10³/g, P＜0.05] in the kidney. Levels of MPO [(1.756±0.283) U/g vs (3.308±0.577) U/g, P＜0.05] and relative expression of TNF - α, IL - 1β, CXCL1, CXCL2 were also lower. Conclusions BLT1 antagonist U75302 protects mice against AKI induced by cisplatin, and the mechanism is associated with reduced infiltration of inflammatory cells in kidney and the inhibition of kidney inflammation.

Objective   To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats.  Methods   The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ, 35 mg/kg) in spontaneous hypertensive rats (SHR). Diabetic rats were divided into diabetic group and the Irbesartan treated group. The pathological changes were investigated by fluorescence microscope and electron microscope. The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose. The concentration of angiotensin II in the supernatant was detected by radioimmunoassay. Immunofluorescence staining was performed to detect the co - localization of CD31 and FSP1.  Results   The significant myocardial fibrosis was presented in the diabetic group. Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats. Double staining of HAEC showed co-localization of CD31 and FSP1, which was decreased by the treatment of Irbesartan (P＜0.05). When HAEC was exposed to high glucose, it showed some cells acquired spindle-shaped morphology and lost CD31 staining, and FSP1 and α - SMA protein expression levels were markedly upregulated, which attenuated by the treatment of Irbesartan.  Conclusion   Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.

Objective    To investigate the inhibition of interstitial fibrosis by NCTD is related to the dephosphorylation of Smad3 linker region mediated by the inhibition of PP2Ac.  Methods   HK -2 cells were cultured and devided into 5 groups: (1) normal control group; (2) TGF-β1 group (5 μg/L); (3) TGF-β1+NCTD group (2.5 mg/L); (4) TGF-β1+PP2Ac shRNA group; (5)TGF-β1+PP2Ac shRNA+ NCTD group. Real-time PCR and Western blot were used to detect the expression of PP2Ac, FN, Col-I, α-SMA and E-cadherin. Additionally, the HK-2 cells were assigned to three groups:(1) normal control group; (2) TGF-β1 group; (3) TGF-β1+NCTD group. Immunofluorescence were used to analysis the distribution of pSmad3-L(Ser²⁰⁴) and pSmad3-L(Ser²⁰⁸). Western blot analysis were used to detect the protein expression of pSmad3-L(Ser²⁰⁴) and pSmad3-L(Ser²⁰⁸).  Results   (1) TGF-β1 stimulated the expression of PP2Ac in HK-2 cells, increased the expression of FN, Col-I and α-SMA, and decreased the expression of E - cadherin. Both NCTD and PP2Ac shRNA could inhibit PP2Ac expression accompanied with the downregulation of FN, Col - I and α - SMA, and upregulation of E - cadherin. However, compared with PP2Ac shRNA transfected group, cells transfecting with PP2Ac shRNA and incubated with NCTD showed no obvious differences on the relief of the above indicators induced by TGF-β1 in HK2 cells. (2)The expression of pSmad3-L(Ser²⁰⁴) and pSmad3-L(Ser²⁰⁸) in the nucleus of HK2 cells stimulated by TGF-β1 was significantly elevated. The expression of pSmad3-L(Ser²⁰⁴) and pSmad3 - L(Ser²⁰⁸) in the nucleus was further upregulated when treated with NCTD.  Conclusions   NCTD has anti - fibrosis effect and it may be due to the inhibition the dephosphorylation of Smad3 linker region mediated by the inhibition of PP2Ac.

Objective    To investigate the effete of chitosan on rabbit carotid artery internal jugular vein fistula intimal hyperplasia and its regulation on TLR4/NF-κB signaling.  Methods    A total of 28 New Zealand white rabbits were randomly divided into the control group(n=4), the model group(n=12) and the chitosan group(n=12). Model group and chitosan group rabbits were established respectively carotid artery internal jugular vein fistula models. After AVF surgery, chitosan was smeared on venous blood vessels and anastomosis. After 4, 6 and 8 weeks, the rabbits were separately sacrificed and the AVF venous vascular tissues were taken. The pathological changes of AVF venous vascular tissue in each group were observed. The changes of α - SMA were detected by immunohistochemistry method. The mRNA expressions of PCNA and TLR4 in the tissues were measured by Real-time PCR. At the same time, the protein expressions of PCNA, TLR4, MyD88 and NF-κB were detected by Western blotting. The experimental data were processed by two-factor analysis of variance in statistics.  Results    (1) After 4 weeks, vascular intimal was thicked in mdel group. In intimal hyperplasia, α-SMA was staining, and then proliferation of vascular smooth muscle cell was significant. As time increasing, more intimal hyperplasia shown obviously , the expression of α-SMA significantly increased. Compared with model group, chitosan group significantly reduced the degree of intimal hyperplasia, the level of α - SMA was significantly decreased,vascular smooth muscle cell proliferation was also extraordinarily decreased. (2) Compared with control group, the expression levels of PCNA, TLR4, MyD88 and NF-κB increased with time. The indices of Chitosan group were markedly higher than control group, but significantly lower than model groups.  Conclusion    Chitosan can inhibit the proliferation of rabbit VSMCs. The mechanism may be concerned in down regulating TLR4- mediated signaling pathway, reducing the possibility of intimal hyperplasia of rabbit AVF venous blood vessels.

Objective    To observe the formation of autophagosome, the expression and distribution of autophagy-related protein LC3-Ⅰ, LC3-Ⅱ and Beclin-1 in adriamycin nephropathy rats at different pathological periods, to explore the relationship between autophagy and renal tissue injury, the occurrence of proteinuria, the progression of renal disease.  Methods   Sixty normal male SD rats were randomly divided into control group (n=30) and model group (n=30), the rats in model group were injected with adriamycin(6.5 mg/kg) via tail - vein for one time, while the rats in control group were injected with saline. Urine protein quantitation of 24 hour, the levels of serum albumin and total cholesterol were measured serially at the 2, 4, 6, 8, 10 weeks. The changes of kidney tissue pathology were detected after HE, PAS and Masson staining by light microscope. The formation of autophagy were detected by transmission electron microscopy, the localization and distribution of LC3-Ⅰ, LC3-Ⅱ and Beclin - 1 were detected by indirect immunofluorescence staining in kidney tissue, the autophagy - related proteins LC3-Ⅰ, LC3-Ⅱ and Beclin-1 expression was detected by Western blotting.  Results    In model group, urinary protein began to increase at the first two weeks, serum albumin decreased at the same time, and total cholesterol increased in the four weeks. There was a statistically significant difference compared with the control group (P＜0.01). The Scr and BUN were increased slightly at the four weeks in model group, and showed the deterioration of renal function after the eight weeks. There was a statistically significant difference compared with the control group (P＜0.01). Mesangial cell proliferation, mitochondrial swelling and foot process broadening and integration appeared early in the model group, while foot process disappearing and nuclear pyknosis were observed in the late by transmission electron microscope; Renal pathology gradually changed from mesangial proliferation to focal segmental glomerulosclerosis (FSGS) by light microscope. A low expression of autophagy was detected in renal tissue of control group rats by transmission electron microscopy and immunofluorescence microscope; in model group, with the progression of disease, the autophagy was significantly enhanced and maintained at a high level. With the progression of disease, the autophagy- related proteins LC3-Ⅰ, LC3-Ⅱ and Beclin-1 was significantly enhanced in the model group than the control group (P＜0.05).  Conclusion    Autophagy is involved in renal tissue injury and the occurrence of proteinuria, closely related to the progression of renal disease.

Objective    To observe the expression of SIRT1 gene and explore its role during the renal development of mice.  Methods    Genotyping was used to observe the impact of SIRT1 knockout on the survival of mice. Immunohistochemistry and immunofluorescence staining were used to examine the location of SIRT1 protein at fetal age of 14 days. Real-time quantitative PCR was used to semi - quantitatively measure the expression level of SIRT1 mRNA in the developing mice kidney at fetal age of 15, 16, 17 and 18 days, and also 0, 5, 9, 23 days after birth. Effects of SIRT1 knockout on size of mice kidney was observed at fetal age of 17, 18 days and also 4, 10 days after birth. Kidneys were cultured in vitro to observe the effects of SIRT1 knockout on ureter bud branching. Immunohistochemical and real - time quantitative PCR were used to observe the effects of SIRT1 knockout on kidney PCNA expression levels.  Results    Most of SIRT1(-/-) mice could still be alive during pregnancy, but died shortly after birth. Immunohistochemistry and immunofluorescence staining showed that the expression of SIRT1 protein could be detected in the early stage of kidney development and could be fully expressed in the metanephric mesenchyme of the developing kidney, but weakly expressed in the ureteric bud. Real - time quantitative PCR analysis revealed that SIRT1 was highly expressed in embryonic kidney at fetal age of 17 days, while the expression level declined quickly thereafter and maintained at low level after birth. Absence of SIRT1 gene resulted in smaller kidney specimens of mice and further microscopic observation showed the glomerular area was smaller and the glomerular number was lesser(P＜0.01). Absence of SIRT1 gene also resulted in decreased ureteral bud branching and declined PCNA expression level of kidney(P＜0.01).  Conclusion    SIRT1 plays an important role in the nephron development in mice by regulating the expression of PCNA.