Objective:To investigate the neuroprotective effect of nicotinamide riboside(NR)on experimental autoimmune en-cephalomyelitis(EAE)mice.Methods:C57BL/6 female mice were induced by myelin oligodendrocyte glycoprotein(MOG35-55)to pro-duce EAE model and were randomly divided into EAE group and NR group.From day 3 to day 27 after immunization,each mouse in EAE group was given normal saline(200 μl/d)and each mouse in NR group was given NR(500 mg/kg,200 μl/d)by intragastric administration.Clinical score and body weight of mice in EAE group and NR group were recorded every day.On the 28th day after immunization,the spinal cord protein of mice in each group was extracted and the frozen sections of spinal cord of mice in each group were prepared.LFB staining was used to detect demyelination,immunofluorescence staining was used to detect the expression of MAP-2 and the number of positive cells of NeuN,BDNF,GDNF,NGF and NT-3,and Western blot was used to detect the expressions of BDNF,GDNF,NGF and NT-3 of spinal cord.Results:Compared with EAE group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05),reduced weight loss,alleviated spinal cord demyelination(P<0.05),increased the expression of MAP-2(P<0.01)and the number of NeuN positive cells(P<0.01),and up-regulated the expressions of BDNF,GDNF,NGF and NT-3(P<0.05).Conclusion:NR shows a good neuroprotective effect on EAE mice.The mechanism may be related to NR significantly increasing the expression of spinal neurotrophic factors,improving the microenvironment of the central nervous sys-tem,nourishing nerves,promoting nerve repair and nerve growth,etc.

Objective:To investigate the neuroprotective effect of nicotinamide riboside(NR)on experimental autoimmune en-cephalomyelitis(EAE)mice.Methods:C57BL/6 female mice were induced by myelin oligodendrocyte glycoprotein(MOG35-55)to pro-duce EAE model and were randomly divided into EAE group and NR group.From day 3 to day 27 after immunization,each mouse in EAE group was given normal saline(200 μl/d)and each mouse in NR group was given NR(500 mg/kg,200 μl/d)by intragastric administration.Clinical score and body weight of mice in EAE group and NR group were recorded every day.On the 28th day after immunization,the spinal cord protein of mice in each group was extracted and the frozen sections of spinal cord of mice in each group were prepared.LFB staining was used to detect demyelination,immunofluorescence staining was used to detect the expression of MAP-2 and the number of positive cells of NeuN,BDNF,GDNF,NGF and NT-3,and Western blot was used to detect the expressions of BDNF,GDNF,NGF and NT-3 of spinal cord.Results:Compared with EAE group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05),reduced weight loss,alleviated spinal cord demyelination(P<0.05),increased the expression of MAP-2(P<0.01)and the number of NeuN positive cells(P<0.01),and up-regulated the expressions of BDNF,GDNF,NGF and NT-3(P<0.05).Conclusion:NR shows a good neuroprotective effect on EAE mice.The mechanism may be related to NR significantly increasing the expression of spinal neurotrophic factors,improving the microenvironment of the central nervous sys-tem,nourishing nerves,promoting nerve repair and nerve growth,etc.

OBJECTIVE T o establish a simple,rapid,highly sensitive and highly specific visualized detection meth-od for Klebsiella pneumoniae based on recombinase polymerase amplification(RPA)technique combined with the lateral flow dipstick(LFD)technique.METHODS The specific primers targeting the khe gene of K.pneumoniae were designed for RPA,the reaction temperature and reaction time for RPA were optimized,the optimal experi-mental conditions for the LFD were determined based on the colorimetric results.The sensitivity,stability,speci-ficity and clinical practicability of RPA-LFD in detection of the K.pneumoniae were evaluated.RESULTS RPA could achieved the optimal detection result for simplification of K.pneumoniae at 34 ℃ for 10 min.The LFD showed the best color development when Milipore HF 135s was used as the nitrocellulose membrane,phosphate buffered saline containing Tween-20(PBST)was used as the sample developing solution,and the concentrations of streptavidin and secondary antibody were 0.8 mg/ml and 0.1 mg/ml,respectively.The detection limit of the RPA-LFD was 5 CFU/ml for detection of K.pneumoniae,the relative standard deviation of the band color inten-sity was less than 5％in six parallel experiments,and there was no cross reactions with other bacterial strains.In addition,the sensitivity of the RPA-LFD in detection of K.pneumoniae was not affected by the serum compo-nents such as proteins and lipids.CONCLUSION The rapid detection method for K.pneumoniae that is established based RPA-LFD is characterized by the simple operation,high sensitivity and high specificity,and it does not need complicated equipment or strict technical requirement for operators and provides a new technique for early diagno-sis and epidemiological survey of the K.pneumoniae.

Objective:Acute kidney injury(AKI) is a common complication after cardiac surgery, and associated with increased risk of development of chronic kidney disease and mortality in the long term. Neutrophil percentage-to-albumin ratio(NPAR) is a new inflammatory marker that has been used to predict the poor prognosis of patients with cardiovascular disease or shock. However, the relationship between NPAR and AKI in patients with cardiac surgery has not been established. The aim was to evaluate the relationship between NPAR and AKI after cardiac surgery.Methods:Data of all adult patients underwent cardiac surgery with cardiopulmonary bypass from January 1, 2006 to December 31, 2018 were extracted from electronic medical record system of the Guangdong Provincial People's Hospital and retrospectively analyzed. The outcome of interest was AKI diagnosed by using the criteria of Kidney Disease Improving Global Outcomes. Logistic regression was used to assess the relationship between NPAR and postoperative AKI while adjust for potential confounders. In addition, restricted cubic spline(RCS) was utilized to provide a flexible description of the association of the preoperative NPAR and AKI. Results:Totally, 24 178 patients were analyzed. The incidence of AKI was 30.1%. Compared with patients without AKI, those with AKI were older and had higher rates of males, left ventricular ejection fraction(LVEF) less than or equal to 0.60, estimated glomerular filtration rate less than 90 ml·min -1·1.73 m -2, hypertension, diabetes, emergency surgery, preoperative critical illness, and reoperation. The baseline serum creatinine, serum uric acid, cardiopulmonary bypass time and postoperative mechanical ventilation time were higher or longer in AKI patients than those in none AKI patients. Then, patients were divided into four groups based on NPAR quartiles. After adjusting for confounding factors using logisitc regression, compared with patients with NPAR in group 3(1.551.97), respectively. The RCS showed a U-shaped nonlinear relationship( P for nonlinear less than 0.001) between NPAR and AKI after adjusting for potential confounding factors. RCS indicated the NPAR of 1.77 was the lowest risk for AKI. When NPAR less than 1.77, per 1-standard deviation(SD) increase in NPAR, the risk of AKI decreased by 23.2%( OR=0.768, 95% CI: 0.687, 0.859). Conversely, when NPAR above 1.77, per one SD increase in NPAR, the risk of AKI increased by 25.9%( OR=1.259, 95% CI: 1.166, 1.36). Conclusion:A U-shaped relationship between NPAR and AKI after cardiac surgery was shown. The appropriate value for NPAR might be 1.77. The risk of AKI might be increased, when less or above this value. NPAR might be used as an effective and economical tool to identify AKI risk among individuals with cardiac surgery, as well as to formulate individualized prevention and intervention strategy.

OBJECTIVE: To investigate the regulatory effect of electroacupuncture (EA) at "Neiguan" (PC6) on mitochondrial autophagy in rats with myocardial ischemia-reperfusion injury (MIRI) at different phases (ischemia and reperfusion phases), and to explore the bidirectional regulatory effects of EA at "Neiguan" (PC6) and its potential mechanism. METHODS: Forty-five male SD rats were randomly divided into 6 groups according to the random number table method, namely, sham-operation group (n=9), model-A group (n=6), model-B group (n=9), EA-A1 group (n=6), EA-B1 group (n=6), and EA-B2 group (n=9). Except the rats in the sham-operation group, the MIRI model was established in the other groups with the physical ligation and tube pushing method. In the model-A group, the samples were collected directly after ligation, and in the model-B group, the samples were collected after ligation and reperfusion. In the EA-A1 group, EA was delivered while the ligation was performed, and afterwards, the samples were collected. In the EA-B1 group, while the ligation was performed, EA was operated at the same time, and after reperfusion, the samples were collected. In the EA-B2 group, during ligation and the opening of the left anterior descending branch of the coronary artery, EA was delivered, and after reperfusion, the samples were collected. EA was performed at bilateral "Neiguan" (PC6), with a disperse-dense wave, a frequency of 2 Hz/100 Hz, a current of 1 mA, and a duration of 30 min. HE staining was employed to observe the morphology of cardiomyocytes, TUNEL was adopted to detect the apoptosis of cardiomyocytes, transcriptome sequencing was to detect the differentially expressed genes in the left ventricle, JC-1 flow cytometry was to detect the mitochondrial membrane potential (MMP) of cardiomyocytes, Western blot was to detect the protein expression of phosphatase and tensin homolog-induced kinase 1 (Pink1), Parkin and p62 in the left ventricle of rats, and ELISA was to detect the levels of serum creatine kinase isoenzyme (CK-MB) and cardiac troponin I (cTn-I) in the rats. RESULTS: Compared with the sham-operation group, the cardiomyocytes of rats in the model-B group were severely damaged, with disordered arrangement, unclear boundaries, broken muscle fibers, edema and loose distribution; and the cardiomyocytes in the EA-B2 group were slightly damaged, the cell structure was partially unclear, the cells were arranged more regularly, and the intact cardiomyocytes were visible. Compared with the sham-operation group, the apoptosis of cardiomyocytes increased in the model-B group (P<0.001); and when compared with the model-B group, the apoptosis alleviated in the EA-B2 group (P<0.001). The differentially expressed genes among the EA-B2 group, the sham-operation group and the model-B group were closely related to cell autophagy and mitochondrial autophagy. Compared with the sham-operation group, MMP of cardiomyocytes was reduced (P<0.001), the protein expression of Pink1, Parkin, and p62 of the left ventricle and the levels of serum CK-MB and cTn-I were elevated in the model B group (P<0.001). In comparison with model-A group, the MMP of cardiomyocytes and the levels of serum CK-MB and cTn-I were reduced (P<0.001, P<0.05), and the protein expression of Pink1 in the left ventricle rose in the EA-A1 group (P<0.01). Compared with the model-B group, MMP of cardiomyocytes increased (P<0.001), the protein expression of Pink1, Parkin, and p62 of the left ventricle, and the levels of serum CK-MB and cTn-I decreased (P<0.001) in the EA-B1 group and the EA-B2 group. When compared with the EA-A1 group, MMP of cardiomyocytes increased (P<0.001), and the protein expression of Pink1, Parkin, and p62 of the left ventricle, and the levels of serum CK-MB and cTn-I decreased in the EA-B1 group (P<0.01). CONCLUSION EA at "Neiguan" (PC6) can ameliorate MIRI in rats, which may be achieved through the Pink1/Parkin-mediated mitochondrial autophagy pathway. EA can alleviate myocardial injury by enhancing mitochondrial autophagy at the ischemia phase, and it can reduce reperfusion injury by weakening mitochondrial autophagy at the reperfusion phase.
Animals ; Electroacupuncture ; Male ; Myocardial Reperfusion Injury/metabolism* ; Rats, Sprague-Dawley ; Rats ; Acupuncture Points ; Autophagy ; Humans ; Mitochondria/genetics*

Objective:Acute kidney injury(AKI) is a common complication after cardiac surgery, and associated with increased risk of development of chronic kidney disease and mortality in the long term. Neutrophil percentage-to-albumin ratio(NPAR) is a new inflammatory marker that has been used to predict the poor prognosis of patients with cardiovascular disease or shock. However, the relationship between NPAR and AKI in patients with cardiac surgery has not been established. The aim was to evaluate the relationship between NPAR and AKI after cardiac surgery.Methods:Data of all adult patients underwent cardiac surgery with cardiopulmonary bypass from January 1, 2006 to December 31, 2018 were extracted from electronic medical record system of the Guangdong Provincial People's Hospital and retrospectively analyzed. The outcome of interest was AKI diagnosed by using the criteria of Kidney Disease Improving Global Outcomes. Logistic regression was used to assess the relationship between NPAR and postoperative AKI while adjust for potential confounders. In addition, restricted cubic spline(RCS) was utilized to provide a flexible description of the association of the preoperative NPAR and AKI. Results:Totally, 24 178 patients were analyzed. The incidence of AKI was 30.1%. Compared with patients without AKI, those with AKI were older and had higher rates of males, left ventricular ejection fraction(LVEF) less than or equal to 0.60, estimated glomerular filtration rate less than 90 ml·min -1·1.73 m -2, hypertension, diabetes, emergency surgery, preoperative critical illness, and reoperation. The baseline serum creatinine, serum uric acid, cardiopulmonary bypass time and postoperative mechanical ventilation time were higher or longer in AKI patients than those in none AKI patients. Then, patients were divided into four groups based on NPAR quartiles. After adjusting for confounding factors using logisitc regression, compared with patients with NPAR in group 3(1.551.97), respectively. The RCS showed a U-shaped nonlinear relationship( P for nonlinear less than 0.001) between NPAR and AKI after adjusting for potential confounding factors. RCS indicated the NPAR of 1.77 was the lowest risk for AKI. When NPAR less than 1.77, per 1-standard deviation(SD) increase in NPAR, the risk of AKI decreased by 23.2%( OR=0.768, 95% CI: 0.687, 0.859). Conversely, when NPAR above 1.77, per one SD increase in NPAR, the risk of AKI increased by 25.9%( OR=1.259, 95% CI: 1.166, 1.36). Conclusion:A U-shaped relationship between NPAR and AKI after cardiac surgery was shown. The appropriate value for NPAR might be 1.77. The risk of AKI might be increased, when less or above this value. NPAR might be used as an effective and economical tool to identify AKI risk among individuals with cardiac surgery, as well as to formulate individualized prevention and intervention strategy.

OBJECTIVE T o establish a simple,rapid,highly sensitive and highly specific visualized detection meth-od for Klebsiella pneumoniae based on recombinase polymerase amplification(RPA)technique combined with the lateral flow dipstick(LFD)technique.METHODS The specific primers targeting the khe gene of K.pneumoniae were designed for RPA,the reaction temperature and reaction time for RPA were optimized,the optimal experi-mental conditions for the LFD were determined based on the colorimetric results.The sensitivity,stability,speci-ficity and clinical practicability of RPA-LFD in detection of the K.pneumoniae were evaluated.RESULTS RPA could achieved the optimal detection result for simplification of K.pneumoniae at 34 ℃ for 10 min.The LFD showed the best color development when Milipore HF 135s was used as the nitrocellulose membrane,phosphate buffered saline containing Tween-20(PBST)was used as the sample developing solution,and the concentrations of streptavidin and secondary antibody were 0.8 mg/ml and 0.1 mg/ml,respectively.The detection limit of the RPA-LFD was 5 CFU/ml for detection of K.pneumoniae,the relative standard deviation of the band color inten-sity was less than 5％in six parallel experiments,and there was no cross reactions with other bacterial strains.In addition,the sensitivity of the RPA-LFD in detection of K.pneumoniae was not affected by the serum compo-nents such as proteins and lipids.CONCLUSION The rapid detection method for K.pneumoniae that is established based RPA-LFD is characterized by the simple operation,high sensitivity and high specificity,and it does not need complicated equipment or strict technical requirement for operators and provides a new technique for early diagno-sis and epidemiological survey of the K.pneumoniae.

Objective:To explore mechanism of Fasudil reducing Aβ1-42 induced BV2 cell injury based on NLRP3 inflamma-some.Methods:BV2 cells were divided into:normal control group,Aβ stimulation group,Aβ+Fasudil intervention group,Aβ+MCC950(NLRP3 inhibitor)intervention group.Cell morphology was observed under microscope.Cell activity was determined of by CCK8.NO release was measured by Griess.NLRP3,caspase 1 and IL-18 expressions were detected by immunofluorescence staining.NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were detected by Western blot.Results:Compared with normal control group,BV2 cells in Aβ stimulation group were activated and showed amoeba-like shape,cell activity was decreased,NO production was increased,NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were increased.Fasudil intervention and MCC950 intervention inhibited cell injury induced by Aβ1-42 in which BV2 cell morphology tended to be normal,cell activity was increased,while produc-tion of NO was reduced,and NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were down-regulated,there was no significant difference between Fasudil intervention group and MCC950 intervention group.Conclusion:Fasudil may alleviate Aβ1-42 induced BV2 cell injury and inflammatory reaction by inhibiting NLRP3 inflammasome activation.

Objective: To explore the effects of protein powder on the bioavailability of perfluoroalkyl substances (PFASs) in blood and kidneys of rats and renal function change. Methods: Twenty-four rats of the SD strain were randomly divided into the negative control group, PFASs group and protein powder group, with 8 rats (half males and half females) in each group. PFASs included 13 perfluorocarboxylic acids (PFCAs) and 8 perfluorosulfonic acids (PFSAs), and the mixture was used as a test subject for intervention. The rats in the negative control group were given deionized water at doses of 20 mL/kg·bw, in the PFASs group were given 5 mL/kg·bw of PFASs mixtures and 15 mL/kg·bw of deionized water, and in the protein powder group were given 5 mL/kg·bw of PFASs mixtures and 15 mL/kg·bw of protein powder (0.258 g/mL). After intervention for 28 successive days, body weight and kidney mass were weighed, and the kidney volume index was calculated. Serum creatinine and blood urea nitrogen were detected by an automatic biochemical analyzer. The PFCAs, PFSAs and PFASs contents were quantified in blood and kidney using ultra-high performance liquid chromatography-electrospray tandem mass spectrometry, and the bioavailability was estimated. Results: There was no significant differences in kidney mass, kidney volume index, serum creatinine and blood urea nitrogen among the negative control group, PFASs group and protein powder group (all P>0.05). The bioavailability of blood PFCAs, PFSAs and PFASs in the protein powder group was not significantly different from the PFASs group (all P>0.05). Compared with the PFASs group, the bioavailability of PFCAs, PFSAs and PFASs were significantly increased in kidneys of male rats in the protein powder group (all P<0.05), while were not significant different in those of female rats (all P>0.05). Conclusion Protein powder at the dose of this study can significantly improve the bioavailability of PFASs in kidneys of male rats, while there no obvious effects on the bioavailability of blood PFASs and renal function.