Deficiency of mindin reduces renal injury after ischemia reperfusion.

BACKGROUND: Acute renal injury (AKI) secondary to ischemia reperfusion (IR) injury continues to be a significant perioperative problem and there is no effective treatment. Mindin belongs to the mindin/F-spondin family and involves in inflammation, proliferation, and cell apoptosis. Previous studies have explored the biological functions of mindin in liver and brain ischemic injury, but its role in AKI is unknown. METHOD: To investigate whether mindin has a pathogenic role, mindin knockout (KO) and wild-type (WT) mice were used to establish renal IR model. After 30 min of ischemia and 24 h of reperfusion, renal histology, serum creatinine, and inflammatory response were examined to assess kidney injury. In vitro, proinflammatory factors and inflammatory signaling pathways were measured in mindin overexpression or knockdown and vector cells after hypoxia/reoxygenation (HR). RESULTS: Following IR, the kidney mindin level was increased in WT mice and deletion of mindin provided significant protection for mice against IR-induced renal injury as manifested by attenuated the elevation of serum creatinine and blood urea nitrogen along with less severity for histological alterations. Mindin deficiency significantly suppressed inflammatory cell infiltration, TNF-α and MCP-1 production following renal IR injury. Mechanistic studies revealed that mindin deficiency inhibits TLR4/JNK/NF-κB signaling activation. In vitro, the expression levels of TNF-α and MCP-1 were increased in mindin overexpression cells compared with vector cells following HR. Moreover, TLR4/JNK/NF-κB signaling activation was elevated in the mindin overexpression cells in response to HR stimulation while mindin knockdown inhibited the activation of TLR4/JNK/ NF-κB signaling after HR in vitro. Further study showed that mindin protein interacted directly with TLR4 protein. And more, mindin protein was confirmed to be expressed massively in renal tubule tissues of human hydronephrosis patients. CONCLUSION: These data demonstrate that mindin is a critical modulator of renal IR injury through regulating inflammatory responses. TLR4/JNK/NF-κB signaling most likely mediates the biological function of mindin in this model of renal ischemia.

Phototherapy with Cancer-Specific Nanoporphyrin Potentiates Immunotherapy in Bladder Cancer.

PURPOSE: Immune checkpoint inhibitors (ICI) in general have shown poor efficacy in bladder cancer. The purpose of this project was to determine whether photodynamic therapy (PDT) with bladder cancer-specific porphyrin-based PLZ4-nanoparticles (PNP) potentiated ICI. EXPERIMENTAL DESIGN: SV40 T/Ras double-transgenic mice bearing spontaneous bladder cancer and C57BL/6 mice carrying syngeneic bladder cancer models were used to determine the efficacy and conduct molecular correlative studies. RESULTS: PDT with PNP generated reactive oxygen species, and induced protein carbonylation and dendritic cell maturation. In SV40 T/Ras double-transgenic mice carrying spontaneous bladder cancer, the median survival was 33.7 days in the control, compared with 44.8 (P = 0.0123), 52.6 (P = 0.0054), and over 75 (P = 0.0001) days in the anti-programmed cell death-1 antibody (anti-PD-1), PNP PDT, and combination groups, respectively. At Day 75 when all mice in other groups died, only 1 in 7 mice in the combination group died. For the direct anti-tumor activity, compared with the control, the anti-PD-1, PNP PDT, and combination groups induced a 40.25% (P = 0.0003), 80.72% (P < 0.0001), and 93.03% (P < 0.0001) tumor reduction, respectively. For the abscopal anticancer immunity, the anti-PD-1, PNP PDT, and combination groups induced tumor reduction of 45.73% (P = 0.0001), 54.92% (P < 0.0001), and 75.96% (P < 0.0001), respectively. The combination treatment also diminished spontaneous and induced lung metastasis. Potential of immunotherapy by PNP PDT is multifactorial. CONCLUSIONS: In addition to its potential for photodynamic diagnosis and therapy, PNP PDT can synergize immunotherapy in treating locally advanced and metastatic bladder cancer. Clinical trials are warranted to determine the efficacy and toxicity of this combination.

Application of Percutaneous Holmium Laser Lithotripsy in the Treatment of Bladder Calculi with Lower Urinary Tract Obstruction or Pelvic Joint Disease.

OBJECTIVE: The aim was to summarize the experience of percutaneous holmium laser lithotripsy in the treatment of bladder calculi with lower urinary tract obstruction or pelvic joint disease in our hospital, explore its efficacy and safety, and improve the minimally invasive surgical technique for bladder calculi. METHODS: The clinical data of 61 patients with bladder calculi combined with lower urinary tract obstructive diseases, including urethral stricture, benign prostatic hyperplasia, and bladder neck contracture or pelvic joint diseases in our hospital from 2017 to 2019 were retrospectively analyzed. All patients with bladder stones measuring 1.5-9 cm were placed in supine or lithotomy position. B-scan was conducted to locate the puncture above the pubic symphysis, establishing a 16-30 Fr bladder channel, and Lumenis holmium laser lithotripsy was subsequently performed through a Li Xun Nephroscope. The crushed stones were flushed out through the percutaneous bladder channel or taken out with foreign body forceps. After surgery, the cystostomy tube was indwelled for 3 days. RESULTS: All the 61 cases were operated successfully with an average lithotripsy time of 25 min, and there was no conversion to open surgery. Postoperative reexamination showed neither residual calculi nor complications such as severe infection, massive hemorrhage, and intestinal injury. CONCLUSION: Percutaneous holmium laser lithotripsy is an improved minimally invasive surgical technique for the treatment of bladder calculi with the advantages of clear surgical field, high stone removal efficiency, less trauma, low-pressure bladder perfusion, and low incidence of accessory injury and infection. For patients with lower urinary tract obstructive disease resulting in obstruction of transurethral surgery and patients with pelvic joint disease resulting in difficult lithotomy position placement, this procedure is more advantageous than transurethral surgery. It is also suitable for bladder calculus with a long diameter >5 cm or multiple calculi.

Combination of cyclin-dependent kinase and immune checkpoint inhibitors for the treatment of bladder cancer.

BACKGROUND: Perturbation of the CDK4/6 pathway is frequently observed in advanced bladder cancer. We investigated the potential of targeting this pathway alone or in combination with chemotherapy or immunotherapy as a therapeutic approach for the treatment of bladder cancer METHODS: The genetic alterations of the CDK4/6 pathway in bladder cancer were first analyzed with The Cancer Genome Atlas database and validated in our bladder cancer patient-derived tumor xenografts (PDXs). Bladder cancer cell lines and mice carrying PDXs with the CDK4/6 pathway perturbations were treated with a CDK4/6 inhibitor palbociclib to determine its anticancer activity and the underlying mechanisms. The combination index method was performed to assess palbociclib and gemcitabine drug-drug interactions. Syngeneic mouse bladder cancer model BBN963 was used to assess whether palbociclib could potentiate anti-PD1 immunotherapy. RESULTS: Of the 413 bladder cancer specimens, 79.2% harbored pertubations along the CDK4/6 pathway. Palbociclib induced G0/G1 cell cycle arrest but with minimal apoptosis in vitro. In mice carrying PDXs, palbociclib treatment reduced tumor growth and prolonged survival from 14 to 32 days compared to vehicle only controls (p = 0.0001). Palbociclib treatment was associated with a decrease in Rb phosphorylation in both cell lines and PDXs. Palbociclib and gemcitabine exhibited antagonistic cytotoxicity in vitro (CI > 3) and in vivo, but palbociclib significantly enhanced the treatment efficacy of anti-PD1 immunotherapy and induced CD8+ T lymphocyte infiltration in syngeneic mouse models. CONCLUSIONS: The CDK4/6 pathway is feasible as a potential target for the treatment of bladder cancer, especially in combination with immunotherapy. A CDK4/6 inhibitor should not be combined with gemcitabine.

Acute Kidney Injury Induced by Pneumoperitoneum Pressure Via a Mitochondrial Injury-dependent Mechanism in a Rabbit Model of Different Degrees of Hydronephrosis.

OBJECTIVE: To clarify the effect of mitochondrial injury during laparoscopic surgery of the kidney in different degrees of hydronephrosis in rabbit model. METHODS: A total of 90 rabbits were randomly allocated into 3 groups (groups PN, PM, and PS, ie, rabbits without, with mild and with severe hydronephrosis, respectively). The rabbits in the PM group (n = 30) and PS group (n = 30) underwent surgical procedures that induced mild and severe left hydronephrosis, respectively. The rabbits in all the groups were then allocated into 5 subgroups and were subjected to intra-abdominal pressures of 0, 6, 9, 12, and 15 mmHg. Changes in the mitochondrial membrane potential and mitochondrial electron microstructure were observed. The apoptosis proteins cytochrome C, apoptosis-inducing factor, caspase-3, and caspase-9 were measured by western blot analysis. RESULTS: As the degrees of hydronephrosis increased, histopathological changes such as the decrease in mitochondrial membrane potential and mitochondrial vacuolization along with increased expression of apoptosis proteins, cytochrome C, apoptosis-inducing factor, caspase-3 gained statistically significance at lower intra-abdominal pressures (In PN and PM groups at 15 mmHg, and in PS group at 9 mmHg; for all P <.01). CONCLUSION: Mitochondrial injury plays an important role during acute kidney injury induced by pneumoperitoneal pressure in different degrees of hydronephrosis in the rabbit model.

[Analysis of flavonoids and antitumor activity of transgenic Saussurea involucrate].

The aim of this paper was to investigate the flavonoids of callus of transgenic and non-transgenic Saussurea involucrate and its antitumor activity on the esophageal cancer CaEs-17 cells. The species and content of mono-phenols were detected by high performance liquid chromatography. The growth of human esophageal cancer CaEs-17 cells was detected using CCK-8 assay, apoptosis morphology observation and flow cytometry. Expression of related apoptotic genes Bax and Bcl-2 were determined by qPCR. The results showed that the content of total flavonoids in the transgenic callus was 2.72 times that of the non-transgenic callus. The cyanidin-galactoside was detected in the transgenic callus, but not in the non-transgenic callus. The inhibitory effect of the extracts from the transgenic callus on CaEs-17 cells was more significant than that of the non-transgenic callus, and the IC50 value had a decreased of 26.4%. Flow cytometry analysis results showed that the apoptosis induction effect of the extracts from the transgenic callus on CaEs-17 cells was significantly better than that of the non-transgenic callus. Fluorescence quantitative PCR analysis results showed that the extracts from the transgenic callus could up-regulate the expression of proapoptotic gene Bax and down-regulate the expression of apoptotic gene Bcl-2, and the regulation effect of the transgenic callus was more significant. Therefore, compared with the non-transgenic callus, the antitumor activity of the extracts from the callus of transgenic S. involucrate on the esophageal cancer CaEs-17 cells was significantly increased, which was closely related to the accumulation of cyanidin-galactoside and its metabolism-related flavonoid compounds in the transgenic callus.

Effect of quantum dots on the biological behavior of the EJ human bladder urothelial carcinoma cell line.

Quantum dots (QDs) are a type of fluorescent label with applications in biological molecules, cells and in vivo imaging. The current study investigated the effect of QDs on the toxicity, proliferation, migration and invasion of the EJ human bladder cancer cell line in vitro. The cell counting kit­8 test was used to measure the survival rate of EJ cells following incubation with varying concentrations of QDs. Additionally, the effect of QDs on tumor cell migration and invasion was evaluated using the Transwell chamber assay, and cell proliferation rate was assessed using a hemocytometer. Data from the current study demonstrated no significant differences in survival rate between the experimental and control groups with the conventionally used concentrations (5, 10 and 20 nM) of QD605 (P>0.05). However, with high concentrations of QD605 (40 and 80 nM), significant differences were observed (P<0.001). The survival rate of EJ cells, however, remained at 92.6%. In addition, no significant differences were observed between the EJ cells labeled with transactivator of transcription (TAT)­QD605 and the unlabeled EJ cells with regard to proliferation, migration and invasion (P>0.05). Thus, the results of the current study indicate that QDs exhibit a certain degree of influence on the activity of the EJ bladder cancer cell line at high concentrations. However, at the concentrations that QDs are conventionally used, there was little impact on the survival of the EJ cells. In addition, the proliferation, migration and invasion abilities of the EJ cells were not affected by TAT­QDs. Therefore, the peptide­conjugated QDs have potential to be applied in the imaging and tracking of live cells in vitro and of animals in vivo. Notably, QDs may provide the foundation for a novel, non­invasive imaging strategy for the early diagnosis of tumors.

Oxidative Damage and Mitochondrial Injuries Are Induced by Various Irrigation Pressures in Rabbit Models of Mild and Severe Hydronephrosis.

OBJECTIVE: We aimed to study whether tolerance to irrigation pressure could be modified by evaluating the oxidative damage of obstructed kidneys based on rabbit models experiencing different degrees of hydronephrosis. METHODS: A total of 66 rabbits were randomly divided into two experimental groups and a control group. In the experimental groups, the rabbits underwent a surgical procedure inducing mild (group M, n=24) or severe (group S, n=24) hydronephrosis. In each experimental group, the rabbits were then randomly divided into 4 subgroups (M0-M3 and S0-S3) consisting of 6 rabbits each. Group 0 received no perfusion. Groups 1 through 3 were perfused with 20, 60 and 100 mmHg fluid, respectively. For the control group, after a sham operation was performed, the rabbits were divided into 4 subgroups and were perfused with fluid at 0, 20, 60 or 100 mmHg of pressure. Kidney injuries was evaluated by neutrophil gelatinase associated lipocalin (NGAL). Oxidative damage was assessed by analyzing superoxide dismutase (Mn-SOD) activity, malondialdehyde (MDA) levels, glutathione reductase (GR), catalase (CAT) and peroxide (H2O2) levels, mitochondrial injuries was assessed by mitochondrial membrane potential (MMP), the mitochondrial ultrastructure and tubular cell apoptosis. RESULTS: In the experimental groups, all results were similar for groups 0 and 1. In group 2, abnormalities were observed in the S group only, and the kidneys of rabbits in group 3 suffered oxidative damage and mitochondrial injuries with increased NGAL, decreased Mn-SOD, GR and CAT,increased MDA and H2O2, lower levels of MMP, mitochondrial vacuolization and an increased apoptotic index. CONCLUSION: In rabbits, severely obstructed kidneys were more susceptible to oxidative damage and mitochondrial injury than mildly obstructed kidneys when subjected to higher degrees of kidney perfusion pressure.

Acute kidney injury induced by various pneumoperitoneum pressures in a rabbit model of mild and severe hydronephrosis.

OBJECTIVE: Increased pneumoperitoneum pressure during laparoscopic surgery can result in acute kidney injury. We aimed to clarify whether intraabdominal pressure tolerance is modified in various degrees of unilateral kidney hydronephrosis in rabbits. METHODS: A total 90 rabbits were randomly allocated to three groups (group PN, PM and PS, i.e. rabbits with no, mild and severe hydronephrosis, respectively, subjected to intraabdominal pressures). Rabbits in group PM (n=30) and group PS (n=30) underwent a surgical procedure inducing a mild or severe left hydronephrosis. Rabbits in all groups were then allocated to 5 subgroups. Then, they were subjected to intraabdominal pressures of 0, 6, 9, 12, and 15 mm Hg, respectively. Acute kidney injury was assessed by measuring serum creatinine (Scr), blood urea nitrogen (BUN), tubular cell apoptosis, kidney injury molecule-1 (KIM-1) and cysteine-rich 61 (Cyr-61/CCN1) expression. RESULTS: Acute kidney injury with increased tubular apoptosis and KIM-1 and Cyr-61 expression occurred when intraabdominal pressure reached 15, 15 and 9 mm Hg in PN, PM and PS groups, respectively. The Scr and BUN levels were similar in all groups. CONCLUSIONS: In rabbits, kidneys with severe hydronephrosis were more likely to suffer acute injury when they were exposed to pneumoperitoneal pressure.

Acute kidney injuries induced by various irrigation pressures in rat models of mild and severe hydronephrosis.

OBJECTIVE: To clarify whether tolerance to irrigation pressure could be modified over varying degrees of kidney obstruction during the endoscopic treatment of kidney stones in a rat model. METHODS: A total of 126 rats were randomly allocated into 2 experimental groups and a control group. The experimental groups underwent a surgical procedure to induce mild (group M, n = 60) or severe (group S, n = 60) hydronephrosis. In each group, the rats were then randomly allocated into 4 subgroups (M0 to M3 and S0 to S3) of respectively 6, 18, 18, and 18 rats. Groups 0 to 3 were respectively perfused with 0 (no irrigation), 20, 60, and 100 mm Hg pressure fluid. The control group underwent no surgical procedures and was only perfused with 100 mm Hg pressure fluid. Acute kidney injuries were assessed by analyzing the kidney microstructure, tubular cell apoptosis, kidney injury molecule-1, and cysteine-rich 61 (Cyr61/CCN1) expression using immunohistochemistry. RESULTS: No abnormalities were observed for the control group, groups 0, or 1. In group 2, abnormalities were observed only in the S group, whereas all kidneys in group 3 suffered acute kidneys injuries, along with occurrence of tubular cells necrosis, increased apoptosis, and increased expression of kidney injury molecule-1 and Cyr61. CONCLUSION: Rats with severely obstructed kidneys were more likely to suffer acute kidney injuries than those with less obstructed kidneys when exposed to higher kidney irrigation pressures. This suggests that the pressure should be controlled and reduced when performing endourologic procedures in the context of kidney obstruction.