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Aim To investigate the molecular mecha-nism of mTORC1/2 inhibitor PP242, which inhibiting cholangiocyte cell preliferation and cystic diliatation via inducing apoptosis and autophagy in the polycystic kid-ney ( PCK ) rats. Methods The expression of p-mTOR and p-Akt in the bile duct epithelial cells was examined by immunohistochemistry. The inhibiting effect of rapamycin and PP242 on cell proliferation ac-tivity on bile duct epithelial cells, the effect of gene si-lence on LC3, Beclin-1 and the effect of the authoph-agy-specific inhibitor 3-methyladenine (3-MA) on cell proliferation were respectively analyzed by WST-1 as-say. The expression of PI3K/Akt signaling pathway re-lated proteins, autophagy-related proteins LC3, Bec-lin-1 and clevead caspase-3, which were treated by PP242 were determined by Western blot. The effect of PP242 on apoptosis was detected by Annexin V/PI double staining and ELISA. The expression of LC3 in cytoplasm was detected by immunofluorescence. The a-bility of rat bile duct epithelial cells spheroid formation was detected by 3D cell culture method, and the cells were treated by single applied with rapamycin and ap- plied rapamycin combined with Rictor gene silencing respectively. Results The protein levels of p-Akt and p-mTOR markedly increased in the bile duct epitheli-um of PCK rats. PP242 inhibited the proliferation of bile duct epithelial cells more effectively than rapamy-cin and showed a dose-and time-dependent manner ( P<0.05 ) . PP242 significantly reduced the levels of PI3K/Akt signaling pathway-related proteins in PCK rat cholangiocytes. PP242 induced apoptosis and auto-phagy, up-regulated the levels of cleaved caspase-3, Beclin-1 and increased the ratio of LC3-II/LC3-I. The combination of Rictor gene silencing and rapamycin was more effective than rapamycin alone in inhibiting cholangiocytes in PCK rats. The inhibitory effect of PP242 on the cell viability was significantly weakened by treatment with 3-MA and knockdown of LC3 and Beclin-1 ( P <0.05 ) . Conclusions PP242 inhibits the proliferation of PCK rat cholangiocytes through PI3K/Akt/mTOR signaling pathway, and the mecha-nism is closely related with autophagy and apoptosis.
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Objective To explore the effects of micro channel (mPCNL)and standard channel percutaneous lithotripsy (sPCNL)on hemodynamics and blood gas analysis in patients with renal calculi.Methods From March 2007 to June 2015,180 patients with kidney stones were selected as the research subjects,through digital said method,the patients were randomly divided into MPCNL group and sPCNL group,the former F16 plastics through established channels of percutaneous nephrolithotomy.The latter F24 plastics through established channels of percuta-neous nephrolithotomy.The changes of hemodynamics and blood gas analysis were observed and compared in the two groups during perioperative period .Results In MPCNL group ,theoperation time was significantly shorter in sPCNL group(t =-6.018),the perfusion fluid was higher than that of sPCNL group(t =20.506,P 0.05).Two groups of postoperative K +,pH,BE,Hb were lower than before anes-thesia,the differences were statistically significant (P <0.05).The two groups of patients with no postoperative urina-ry sepsis and other serious complications.Conclusion With the increase of operation time filling fluid absorption on hemodynamics and arterial blood gas analysis of influence gradually increased;in heart,lung and normal renal function patients,due to the compensatory organ function,caused by MPCNL and sPCNL irrigation fluid absorption differences in the amount is not enough to cause different hemodynamics and blood gas analysis.
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AIM: To investigate the roles of angiotensin Ⅱ and NADPH oxidase in the development of renal oxidative stress (OS) in a rat model of hyperoxaluria. METHODS:Animal model of hyperoxaluria was established in a-dult male Sprague - Dawley rats by administration of 0.8% ethylene glycol (EC) in drinking water for 4 weeks. Simultaneous treatment with apocynin (0.2g·kg~(-1)·d~(-1))or losartan (30 mg·kg~(-1)·d~(-1) ) by intragastric administration were performed in rats, respectively. At the end of the study, markers for the state of oxidative stress (OS) , urinary 8 - IP and the enzymatic activity of superoxide dismutase ( SOD) in kidney homogenates were assessed. The concentration of angiotensin H in kidney homogenates was determined using radioimmunoassay method. Expression of NADPH oxidase subunit p47phox in kidney was localized and evaluated by immunohistochemistry and real time - PCR, respectively. RESULTS: p47phox expressed widely in the kidneys of this rat model, including renal cortex, inner medulla and outer medulla. Compared with the control, OS developed significantly in rats received EG, with increased expression of p47phox mRNA in kidneys. Renal angiotensin Ⅱ also increased significantly. Treatment with apocynin or losartan significantly reduced the excretion of urinary 8 - IP, restored the SOD activity, with decrease in the expression of p47phox mRNA in kidney, but the levels of those OS markers in apocynin or losartan treated rats were still higher than those in normal controls. CONCLUSION: Results suggest that renal Ang Ⅱ and its stimulation of NADPH oxidase may partially account for the development of OS in kidney in this rat model of hyperoxaluria.
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Objective To explore the key points of the diagnosis and treatment of severe pneumonia following renal transplantation in the elderly. Methods The clinical data of 28 patients with severe pneumonia following renal transplantation were retrospectively analyzed, including 20 cases aged <60 years (<60 years old group) and 8 cases aged ≥60 years (≥60 years old group). Results In <60 years old group, the severe pneumonia occurred during 1-13 months after the renal transplantation. All the patients had fever. 10 cases coughed and 8 cases had expectoration. 6 cases had type I respiratory failure (RF) and 3 cases experienced type 11 RF. 6 cases had lobar pneumonia and 13 cases occurred interstitial pneumonia. One case experienced lung consolidation. The pathogens of 16 cases in <60 years old group were identified, including 4 cases with bacterial pneumonia, 4 cases with cytomegalovirus (CMV) pneumonia, 2 cases with pneumocystis carinii pneumonia, Ⅰ case with mycoplasma infection, Ⅰ case with tuberculosis infection, and 4 cases with mixed infection (2 cases infected by bacteria plus CMV, 1 case by bacteria plus fungi and 1 case by bacteria plus tuberculosis). Combined drugs (broad-spectrum antibiotic, antivirus and antifungal agent) were administered on the initial stage and sensitive drugs were used later according to the pathogens. Hormone or immunoglobulin was used when other drugs were useless. 17 cases were cured and 3 cases died. In ≥60 years old group, the severe pneumonia occurred during 1-9 months after renal transplantation. All 8 patients had fever, too. 5 cases coughed and 3 cases had expectoration. 3 cases experienced type ⅠRF and 1 case experienced type Ⅱ RF. 3 cases had lobar pneumonia and 5 casesoccurred interstitial pneumonia. The pathogens of 5 eases were identified. Among them, 2 cases were affected by bacterial pneumonia, 1 case by CMV pneumonia and 2 cases by mixed pneumonia (one by bacteria plus CMV, one by bacteria plus fungi). Similar modality was applied, and 5 cases were cured and 3 cases died. Conclusions Most of severe pneumonia occur during 1-9 months after renal transplantation in the elderly. The main pathogens are bacteria and CMV. Medications for all of the most common pathogens and assisted ventilation should be used early. Specific narrow-spectrum antibiotic or antiviral drugs could be used quickly after pathogens were identified, and hormone or immunoglobulin could be administered to patients when the infection is severe or the pathogens are uncertain.