Exosomes from miR-23 Overexpressing Stromal Cells Suppress M1 Macrophage and Inhibit Calcium Oxalate Deposition in Hyperoxaluria Rat Model.

Purpose: To investigate whether ADSC-derived miR-23-enriched exosomes could protect against calcium oxalate stone formation in a hyperoxaluria rat model. Methods: An ethylene glycol- (EG-) induced hyperoxaluria rat model and an in vitro model of COM-induced HK-2 cells coculturing with RAW264.7 cells were established to explore the protective mechanisms of ADSC-derived miR-23-enriched exosomes. Results: The results showed that treatment with miR-23-enriched exosomes from ADSCs protected EG-induced hyperoxaluria rats, and cell experiments confirmed that coculturing with miR-23-enriched exosomes alleviated COM-induced cell autophagy. Overexpressed miR-23 suppressed M1 macrophage polarization by inhibiting IRF1 expression. Furthermore, the predicted binding site between the IRF1 messenger RNA 3'-untranslated region (3'-UTR) and miR-23 was confirmed by the dual-luciferase reporter assay. Conclusion: In conclusion, our research gave the first evidence that ADSC-derived miR-23-enriched exosomes affected the polarization of M1 macrophages by directly inhibiting IRF1 and protecting against calcium oxalate stone formation in a hyperoxaluria rat model.

Cystine crystal-induced reactive oxygen species associated with NLRP3 inflammasome activation: implications for the pathogenesis of cystine calculi.

PURPOSE: To investigate whether cystine crystal-induced production of reactive oxygen species (ROS) and activation of NLRP3 inflammasome contribute to cystine calculi formation. METHODS: Slc7a9-knockout rats were created as cystine calculi animal models. Kidney histological examination using TEM and immunohistochemistry were performed. The protein expression of NLRP3 and IL-1ß and the concentrations of oxidative stress markers such as ROS, MDA and H2O2 in kidney tissues were estimated. In parallel, HK-2 human renal proximal tubule cells were exposed to cystine crystals and NAC treatment. The protein and mRNA expression levels of NLRP3 were evaluated. Finally, cell apoptosis and cystine crystal adherence were also assessed. RESULTS: Activation of the NLRP3 inflammasome and marked elevations in MDA, H2O2 and ROS levels were observed both in vivo and in vitro. In particular, the protein and mRNA expression of NLRP3 was significantly increased by cystine crystals, but could be restored by an inhibitor of ROS. In addition, cell apoptosis and cystine crystal adherence were promoted by the NLRP3 inflammasome. The expression of CD44, OPN and HA in HK-2 cells was markedly increased by cystine crystals, but could be decreased by NLRP3 siRNA treatment. CONCLUSION: Notably, we found that the activation of NLRP3 by cystine crystal-induced ROS production was of major importance in the pathogenesis of cystine calculi formation.

Ceftriaxone Calcium Crystals Induce Acute Kidney Injury by NLRP3-Mediated Inflammation and Oxidative Stress Injury.

OBJECTIVE: To investigate the role of inflammatory reactions and oxidative stress injury in the mechanisms of ceftriaxone calcium crystal-induced acute kidney injury (AKI) both in vivo and in vitro. METHODS: Male Sprague Dawley rats were randomly divided into five groups of ten each according to different concentrations of ceftriaxone and calcium. Based on the levels of serum creatinine (Scr) and blood urea nitrogen (BUN), the AKI group was chosen for the subsequent experiments. Kidney histological examination and immunohistochemistry were performed. The expression of NLRP3 and IL-1ß protein and the concentrations of oxidative stress markers such as ROS, MDA, and H2O2 in kidney tissues were estimated. In parallel, HK-2 human renal proximal tubule cells were exposed to ceftriaxone calcium crystals. The mRNA expression levels of NLRP3 and IL-1ß and the concentrations of oxidative stress markers were evaluated. Finally, cell viability and rat survival were also assessed. RESULTS: The results showed that significantly increased Scr and BUN levels, consistent with morphological changes and kidney stones, were found in the rats that received the highest concentration of ceftriaxone (1000 mg/kg) combined with calcium (800 mg/kg). The activation of the NLRP3 inflammasome axis and the marked elevation of MDA, H2O2, and ROS levels were observed both in vivo and in vitro. High expression of Nrf2, HO-1, and NQO1 was also documented. In addition, cell apoptosis and rat mortality were promoted by ceftriaxone calcium crystals. CONCLUSIONS: Notably, we found that ceftriaxone-induced urolithiasis was associated with a high risk of AKI and NLRP3-mediated inflammasome and oxidative stress injury were of major importance in the pathogenesis.

Protective effect of salvianolic acid B against oxidative injury associated with cystine stone formation.

The aim of this study was to investigate the role of oxidative stress in cystine crystal formation and whether salvianolic acid B, a natural antioxidant, could prevent cystine-mediated oxidative injury in vivo and in vitro. The levels of oxidative stress and antioxidase activity in cystine stone patients were assessed. Then, the oxidative stress exerted by cystine on human kidney-2 (HK-2) cell viability and biochemical parameters including antioxidase activity and antioxidant protein expression were evaluated, and the protective action of salvianolic acid B was also examined. Finally, salvianolic acid B was tested to determine whether it could prevent or reduce renal crystal formation in Slc7a9 knockout mice. The activity levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were decreased, and the amount of malondialdehyde (MDA) was increased in patients with cystine stones compared with people without cystine stones (p < 0.05). Significant reductions in cell viability, antioxidase activity and antioxidant protein expression levels were found in the cystine group compared with controls. However, such oxidative injuries were prevented by salvianolic acid B. In the animal study, loose crystals with white spots were seen in the renal parenchyma, bilateral renal pelvis and bladders in the Slc7a9 knockout group. In contrast, no renal crystals were seen in the control group, and markedly fewer crystals with significantly higher antioxidase activity and diminished oxidative stress were detected in the salvianolic acid B group. Cystine cytotoxicity in vitro and cystine stone formation in vivo were associated with oxidative stress, and salvianolic acid B could protect against cystine stone-induced injury.

Clinical value of 68Ga-PSMA-11 PET/CT in the diagnosis of castration-resistant prostate cancer / 中华核医学与分子影像杂志

Objective To assess the role of 68Ga-N,N′-bis(2-hydroxy-5-(carboxyethyl)benzyl) ethylenediamine-N,N′-diacetic acid(HBED-CC)-(Ahx)Lys-CO-Glu(PSMA-11) PET/CT on the detection of metastatic lesions from castration-resistant prostate cancer (CRPC).Methods Sixteen patients with CRPC who underwent 68Ga-PSMA-11 PET/CT between January 2015 and November 2015 were recruited in this study.Mean age of patients was (72±9) years.The PSA levels were 4-12 356 μg/L, Gleason score was 7-10.PET/CT was performed at 1 h postinjection of 68Ga-PSMA-11.Patient-based analysis and lesion-based analysis were performed.ROI analysis was used to calculate the tumor uptake (SUVmax).Final diagnosis was based on histopathology and results of other imaging examinations(99Tcm-MDP imaging, MRI).χ2 test was used to compare the diagnostic efficiencies of PET and CT.Results No adverse effects were observed in patients.68Ga-PSMA-11 PET/CT showed moderate physiologic uptake in salivary glands and proximal small intestine, with predominant tracer clearance by the kidneys.All patients were positive on 68Ga-PSMA-11 PET/CT.Bone metastasis was found in 16 patients, liver metastasis in 2 patients (5 lesions), and lymph node metastasis in 4 patients (26 lesions).The SUVmax of liver, lymph node and bone metastases were 15.06±2.77, 7.54±5.20, 19.01±16.96, respectively.The diagnostic sensitivity, specificity and accuracy on bone metastasis with 68Ga-PSMA-11 PET and CT were 96.30%(52/54) vs 61.11%(33/54), 3/3 vs 1/3, 96.49%(55/57) vs 59.65%(34/57).The sensitivities and accuracies of the two modalities were significantly different(χ2=19.943, 22.593, both P<0.01).Conclusions 68Ga-PSMA-11 PET/CT could precisely detect both primary and metastatic lesions of CRPC, suggesting that it is of great value for the clinical management and treatment.

Clinical value of 68Ga-PSMA-11 PET/CT in the diagnosis of castration-resistant prostate cancer / 中华核医学与分子影像杂志

Objective To assess the role of 68Ga-N,N'-bis (2-hydroxy-5-(carboxyethyl) benzyl) ethylenediamine-N,N'-diacetic acid(HBED-CC)-(Ahx) Lys-CO-Glu(PSMA-11) PET/CT on the detection of metastatic lesions from castration-resistant prostate cancer (CRPC).Methods Sixteen patients with CRPC who underwent 68Ga-PSMA-11 PET/CT between January 2015 and November 2015 were recruited in this study.Mean age of patients was (72±9) years.The PSA levels were 4-12 356 μg/L,Gleason score was 7-10.PET/CT was performed at 1 h postinjection of 68Ga-PSMA-11.Patient-based analysis and lesionbased analysis were performed.ROI analysis was used to calculate the tumor uptake (SUVmax).Final diagnosis was based on histopathology and results of other imaging examinations(99Tcm-MDP imaging,MRI).x2 test was used to compare the diagnostic efficiencies of PET and CT.Results No adverse effects were observed in patients.68Ga-PSMA-11 PET/CT showed moderate physiologic uptake in salivary glands and proximal small intestine,with predominant tracer clearance by the kidneys.All patients were positive on 68Ga-PSMA-11 PET/CT.Bone metastasis was found in 16 patients,liver metastasis in 2 patients (5 lesions),and lymph node metastasis in 4 patients (26 lesions).The SUVmax of liver,lymph node and bone metastases were 15.06±2.77,7.54±5.20,19.01± 16.96,respectively.The diagnostic sensitivity,specificity and accuracy on bone metastasis with 68Ga-PSMA-11 PET and CT were 96.30％(52/54) vs 61.11％(33/54),3/3 vs 1/3,96.49％ (55/57) vs 59.65％ (34/57).The sensitivities and accuracies of the two modalities were significantly different(x2=19.943,22.593,both P＜0.01).Conclusions 68Ga-PSMA-11 PET/CT could precisely detect both primary and metastatic lesions of CRPC,suggesting that it is of great value for the clinical management and treatment.

Research progress and prospect of decellularized matrix in kidney tissue engineering / 中国组织工程研究

BACKGROUND: Previous studies have demonstrated that the well combination of decellularized matrix and appropriate seeded cells could construct a tissue-engineered kidney.OBJECTIVE: To review advances in kidney tissue engineering and decellularized matrix.METHODS: The first author retrieved CNKI, Wanfang and PubMed databases for articles addressing kidney tissue engineering published from January 1996 to April 2016. The key words were decellularized matrix,extracellular matrix,tissue engineering, kidney, seeded cells in English and Chinese,respectively.RESULTS AND CONCLUSION: The decellularized matrix loses its immunogenicity due to the removal of cellular components, while it retains the important bioactive components of the extracellular matrix and the ultrastructure of the tissues and organs, making it more and more important in kidney tissue engineering. The decellularized matrix especially exerts an important role in the tissue-engineered construction of the entire kidney as driven by recently emerging all-organ acellular cell technology. Remarkable advances in kidney tissue engineering and decellularized matrix have been made in recent years, and realized the construction of a certain functional tissue-engineered kidney. However, there are still many challenges on the way to construct a completely functional tissue-engineered kidney.BACKGROUND: Previous studies have demonstrated that the well combination of decellularized matrix and appropriate seeded cells could construct a tissue-engineered kidney.OBJECTIVE: To review advances in kidney tissue engineering and decellularized matrix.METHODS: The first author retrieved CNKI, Wanfang and PubMed databases for articles addressing kidney tissue engineering published from January 1996 to April 2016. The key words were decellularized matrix,extracellular matrix,tissue engineering, kidney, seeded cells in English and Chinese,respectively.RESULTS AND CONCLUSION: The decellularized matrix loses its immunogenicity due to the removal of cellular components, while it retains the important bioactive components of the extracellular matrix and the ultrastructure of the tissues and organs, making it more and more important in kidney tissue engineering. The decellularized matrix especially exerts an important role in the tissue-engineered construction of the entire kidney as driven by recently emerging all-organ acellular cell technology. Remarkable advances in kidney tissue engineering and decellularized matrix have been made in recent years, and realized the construction of a certain functional tissue-engineered kidney. However, there are still many challenges on the way to construct a completely functional tissue-engineered kidney.

Erythropoietin mobilizes renal progenitor cells to reduce ischemic reperfusion injury in rats with nephron-sparing surgery / 中华肾脏病杂志

Objective To investigate the effects of the erythropoietin (EPO) on ischemia reperfusion injury (IRI) in rats with nephron-sparing surgery (NSS). Methods Fifty-four Sprague Dawley rats were divided into 3 groups randomly after right kidney nephrectomy: Sham group, NSS group (PBS+NSS) and EPO group (EPO+NSS). During NSS, renal artery was clamped for 40 min to induce IRI. Sham group just adopted exposure renal artery without vascular clamped. Rats in NSS group were injected intraperitoneally with PBS for 3 days before NSS. Rats in EPO group were injected intraperitoneally with EPO for 3 days before NSS. After 12 h, 24 h, 72 h, blood sample and renal tissues were collected. The serum creatinine (Scr) and urea nitrogen (BUN) were evaluated. The pathology injury was evaluated by HE staining. The CD24/CD133 double-positived renal progenitor cells (RPCs) were tested by flow cytometry. The CD133 and PCNA protein were quantified by immunohistochemical staining. The expressions of Wnt7b and β-catenin protein were detected by Western blotting. Results Rats in NSS group had more elevated Scr, BUN and pathology injury scores 12 h, 24 h and 72 h after operation than those in Sham group (all P<0.05). Compared with those in the NSS group, the Scr and BUN in the EPO group were significantly lower 24 h after the surgery (all P<0.05), and the pathology injury score also decreased (P<0.05). The proportion of RPCs, expressions of CD133 and PCNA, and expressions of Wnt7b and β-catenin protein were significantly higher after 24 h of the surgery in NSS group than those in the Sham group (all P<0.05). While compared with those in the NSS group, the proportion of RPCs and expressions of CD133, PCNA, Wnt7b and β-catenin increased at the EPO group (all P<0.05). Conclusions EPO can reduce the IRI after NSS, and its mechanism may be related to the mobilization of the RPCs by the Wnt7b/β-catenin signal pathway.

Sunitinib as first-line neoadjuvant therapy in two patients with rectal stromal tumors.

In the treatment of rectal stromal tumors, which account for approximately 5% of gastrointestinal stromal tumors, molecular-targeted neoadjuvant therapy should be considered if the tumor is too large to achieve R0 grade resection or multiple visceral resection is required. Currently, imatinib is generally recommended as the first-line agent for such therapy. Although it has been reported that neoadjuvant therapy in patients experiencing imatinib resistance or intolerable adverse events can be successfully achieved by switching to sunitinib, first-line use of sunitinib for neoadjuvant therapy of gastrointestinal stromal tumors has not previously been reported. In this case report, first-line sunitinib neoadjuvant therapy of two patients who had very large rectal stromal tumors at sites close to the prostate and bladder produced good clinical outcomes.

Impact of ischemic preconditioning on dynamics of homing of endothelial progenitor cells after renal ischemia reperfusion injury / 中华肾脏病杂志

Objectives To investigate the impact of ischemic preconditioning (IPC) on dynamics of homing of endothelial progenitor cells (EPCs) after renal ischemia reperfusion injury (IR).Methods Sixty male Sprague-Dawley rats were randomly divided into three groups after right-side kidney nephrectomy:for sham-operated rats,lumbotomy without vascular clamping was performed; IR rats were clamped renal blood vessels for 40 minutes while IPC rats were pre-treated with 15 min ischemia and 10 min reperfusion.At 3,12,24 h,and 3 days after reperfusion,the pool of circulating,kidneys,lungs and spleens were harvested.The extent of renal injury was assessed by biochemical and histological examination.The dynamics of homing of EPCs was observed by flow cytometry.Results The rats in IPC group exhibited significant improvements in renal function and morphology.Compared with IR group and sham group,the number of EPCs in blood was increased in the IPC group at 12 h and 24 h after reperfusion (P ＜ 0.05).The number of EPCs in kidney was increased at all times pointin the IPC group and IR group as compared to the sham group (P ＜ 0.05.In addition,EPCs number was increased in IPC group compared with the IR group at 12 h and 24 h [(11.36±0.66)％ vs (6.37±0.69)％,(6.31±0.70)％ vs (4.40±0.60)％,all P＜ 0.05].Compared with IR group and sham group,the number of EPCs in the lung was increased in the IPC groups at 12 h after reperfusion [(2.95±0.66)％ vs (1.78±0.59)％,(1.66±0.61)％,all P ＜ 0.05].The number of EPCs in spleen was increased in the IPC group at 72 h as compared with the IR group and sham group [(0.55±0.06)％ vs (0.34±0.07)％,(0.31±0.06)％,all P ＜ 0.05].Conclusions Endogenous EPCs may home to injured kidney after IPC.EPCs can also gather in the lungs and spleen.

New progress of Rapamycin in prevention and treatment of chronic allograft nephropathy / 医学研究生学报

Chronic allograft nephropathy(CAN) represents the primary cause of renal graft loss.Calcineurin inhibitor(CI) nephrotoxicity may be one of the most important pathogenetic factors in the development of this CAN.Rapamycin(Rapa)is a new immunosuppressive drug,its mechanism is different from that of CI and without nephrotoxicity.Moreover,Rapa has shown potent antiproliferative activity in different clinical and experimental models.The author reviewed relevant study of Rapa on preventing and treatment CAN.

The clinical significance of microvascular invasion to the neogrowth in clinically nonmetastatic renal cell carcinoma / 中华泌尿外科杂志

Objective To study the significance of microvascular invasion to the neogrow in clinically nonmetastatic renal cell carcinoma (RCC). Methods Between 1989 and 1996,70 patients (mean age 54) were followed up for 1 to 7 years after radical nephrectomy for clinically localized RCC. Among them,there were 5 PT 1,50 PT 2,14 PT 3 and 1 PT 4 and histologically 7 G 1,38 G 2,19 G 3 and 6 G 4.The mean tumor diameter was 7.2 cm.The slides were stained with hematoxylin and eosin,elastin stains and periodic acid Schiff. then,the presence or absence of clinically inapparent vascular invasion,the relevance of microscopic vascular invasion to conventional tumor stage,grade and tumor diameter,et al were studied. Results Of the 70 patients analyzed,24(34.3%) had microvascular invasion,while 46 had none on microscopic examination.Of the microscopic vascular invasion group (11/24,45.8%),7 subsequently died of cancer recurrence,2 noncancer related death,4 alive with metastatic disease while only 4/46(8.7%) without microscopic vascular invasion presented with disease progression.Chi-square test showed statisticaly significant difference between stage,grade,tumor diameter and presence or absence of microscopic vascular invasion. A multivarite analysis was performed considering the impacts of age,PT stage,tumor grade,tumor diameter and microscopic vascular invasion on disease progression,an increase in statistical significance was confirmed with Coxs proportional hazards model(P=0.0062,RR=0.378). So, microscopic vascular invasion seems to be the most important predictor of progression in RCC. Conclusions In patients underwent radical nephrectomy for clinically nonmetastatic RCC,microvascular invasion may be another important prognostic marker and this makes us considering the presence of microscopic vascular invasion in RCC might be another pathological subcategory to predict the prognosis of RCC and can be used to choose whether early adjuvant therapy is necessary.