Objective: To investigate the effects of mono-carbonyl analogues of curcumin (L6H21) on paraquat (PQ) -induced injury in HK-2 cell line and explore its underlying mechanisms. Methods: Cultured HK-2 cells were challenged by PQ with or without L6H21 treatment. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry, respectively. Gene expressions and protein levels of apoptotic and inflammatory factors were assessed by RT-PCR, ELISA, and western blot. Intracellular ROS production was detected by DCFH-DA staining. Superoxide dismutase (SOD) and malondialdehyde (MDA) were examined by chemical colorimetry. Results: 1) PQ challenge significantly inhibited HK-2 cells proliferation, which was prevented by L6H21 administration. PQ dramatically induced HK-2 apoptosis evidenced by increasing expressions of caspase-9, caspase-3 and Bax, while decreasing Bcl-2 level. However, PQ induced these apoptotic effects in HK-2 cells were reversed by L6H21. Similarly, PQ exposure obviously enhanced activity of NF-κB and levels of cytokines (TNF-α、IL-6) in HK-2 cells, which was inhibited by L6H21. Furthermore, administration of L6H21 inhibited PQ induced ROS and MDA production, and promoted SOD level in HK-2 cells. Conclusion L6H21 administration inhibits PQ-induced apoptosis in HK-2 cells possibly by reducing inflammation and oxidative damage.

Objective:To investigate the protective effect of somatostatin (SS) on acute kidney injury (AKI) of paraquat (PQ) poisoned mice and its mechanism.Methods:From December 2017 to April 2018, a total of 48 SPF male BALB/C mice were selected and randomly divided into 4 groups, with 12 mice in each group: Control group, SS group (20 mg/kg SS was injected 1 hour before and 3 hours after gavage with normal saline) , PQ group (2% PQ 60 mg/kg by gavage) and PQ+SS group (Intragastric administration was performed with 2% PQ solution of 60 mg/kg, and 20 mg/kg SS was administered 1 h before and 3 h after intragastric administration) , 12 mice in each group were observed for the general situation and behavioral effects. After 24 hours of modeling, mice were sacrificed.Then blood was extracted after eyeball was removed, and both kidneys were removed by laparotomy. Serum IL-6, TNF-α and MPO levels were determined by ELISA. The characteristic pathological changes of toxic renal tubular injury were observed under light microscope and scored accordingly. The changes of NF-κB expression were detected by Western-Blot, SOD, Caspase-3 and malondialdehyde (MDA) were detected by chemical colorimetry.Results:Mice in Control group and SS group showed normal general conditions and behaviors; Mice in PQ group were significantly worse than those in Control group, showing decreased feeding and activity, dry fur, hair shedding and listless spirit; The above symptoms in the mice of PQ+SS group were alleviated compared with the PQ group. Under the light microscope, the renal tissue structure of PQ group was obviously disordered and severely damaged, and the nephropathy score was (6.14±0.72) . The performance of PQ+SS group under light microscope was improved compared with PQ group, and nephropathy score (4.36±0.42) decreased ( P<0.05) . Compared with the Control group, serum TNF-α (39.89±3.32) pg/ml, IL-6 (77.29±4.77) pg/ml, renal NF-κB (2.29±0.097) , MPO (0.31±0.017) μg/ml, MDA (0.91±0.03) mmol/mg prot, and Caspase-3 (376.51±8.24) % levels were significantly increased in the PQ group, while the level of renal SOD (2.36±0.73) U/mg prot was significantly decreased ( P<0.05) . Compared with the PQ group, serum TNF-α (33.82±1.57) pg/ml, IL-6 (58.49±5.89) pg/ml, renal NF-κB (0.84±0.05) , MPO (0.22±0.01) μg/ml, MDA (0.72±0.05) mmol/mg prot, Caspase-3 (327.32±21.93) % decreased significantly, and renal SOD (4.90±0.81) U/mg prot increased significantly in the PQ+SS group ( P<0.05) . Conclusion:PQ poisoning can lead to AKI in mice, while SS can reduce AKI caused by PQ poisoning, improve the general survival state of PQ poisoned mice, and play a certain protective role in kidney injury caused by PQ poisoning, which may be achieved by inhibiting oxidative stress response, inflammatory response and apoptosis caused by poisoning.

Objective:To investigate the protective effect of somatostatin (SS) on acute kidney injury (AKI) of paraquat (PQ) poisoned mice and its mechanism.Methods:From December 2017 to April 2018, a total of 48 SPF male BALB/C mice were selected and randomly divided into 4 groups, with 12 mice in each group: Control group, SS group (20 mg/kg SS was injected 1 hour before and 3 hours after gavage with normal saline) , PQ group (2% PQ 60 mg/kg by gavage) and PQ+SS group (Intragastric administration was performed with 2% PQ solution of 60 mg/kg, and 20 mg/kg SS was administered 1 h before and 3 h after intragastric administration) , 12 mice in each group were observed for the general situation and behavioral effects. After 24 hours of modeling, mice were sacrificed.Then blood was extracted after eyeball was removed, and both kidneys were removed by laparotomy. Serum IL-6, TNF-α and MPO levels were determined by ELISA. The characteristic pathological changes of toxic renal tubular injury were observed under light microscope and scored accordingly. The changes of NF-κB expression were detected by Western-Blot, SOD, Caspase-3 and malondialdehyde (MDA) were detected by chemical colorimetry.Results:Mice in Control group and SS group showed normal general conditions and behaviors; Mice in PQ group were significantly worse than those in Control group, showing decreased feeding and activity, dry fur, hair shedding and listless spirit; The above symptoms in the mice of PQ+SS group were alleviated compared with the PQ group. Under the light microscope, the renal tissue structure of PQ group was obviously disordered and severely damaged, and the nephropathy score was (6.14±0.72) . The performance of PQ+SS group under light microscope was improved compared with PQ group, and nephropathy score (4.36±0.42) decreased ( P<0.05) . Compared with the Control group, serum TNF-α (39.89±3.32) pg/ml, IL-6 (77.29±4.77) pg/ml, renal NF-κB (2.29±0.097) , MPO (0.31±0.017) μg/ml, MDA (0.91±0.03) mmol/mg prot, and Caspase-3 (376.51±8.24) % levels were significantly increased in the PQ group, while the level of renal SOD (2.36±0.73) U/mg prot was significantly decreased ( P<0.05) . Compared with the PQ group, serum TNF-α (33.82±1.57) pg/ml, IL-6 (58.49±5.89) pg/ml, renal NF-κB (0.84±0.05) , MPO (0.22±0.01) μg/ml, MDA (0.72±0.05) mmol/mg prot, Caspase-3 (327.32±21.93) % decreased significantly, and renal SOD (4.90±0.81) U/mg prot increased significantly in the PQ+SS group ( P<0.05) . Conclusion:PQ poisoning can lead to AKI in mice, while SS can reduce AKI caused by PQ poisoning, improve the general survival state of PQ poisoned mice, and play a certain protective role in kidney injury caused by PQ poisoning, which may be achieved by inhibiting oxidative stress response, inflammatory response and apoptosis caused by poisoning.

Objective To characterize the biomarkers of urine samples for early diagnosis of colorectal cancer(CRC)using proton nuclear magnetic resonance (1H-NMR)combined with pattern recognition.Methods 400 MHz 1H-NMR was used to test the urine samples obtained from 23 patients with Ⅰ/Ⅱ stage CRC,40 healthy controls (HC)and 18 patients with esophageal cancer (EC). Pattern recognition through orthogonal partial least squares-discriminant analysis (OPLS-DA)was applied on 1H-NMR data to find urine metabolic differences between CRC and HC.Results OPLS-DA could effectively determine HC,patients withⅠ/Ⅱstage CRC and patients with esophageal cancer.Compared with HC,early stage CRC had significant decreases of choline,isocitric acid,lactamine,phenylalan, cysteine,creatinine,aspartic acid,hippurate acid,methylamine,dimethyl sulfone,and increases of acetoacetate,glutamine,glycocyamine,cis-aconitate, trans-aconitate,homocycteine in the urine samples.Conclusion Urine metabonomics based on NMRIndicates that glucose metabolism,amino acid metabolism,choline metabolism,energy metabolism and intestinal microflora are disturbance in colorectal cancer patients,which provide valuable metabolic information on the molecular level for early diagnosis of colorectal cancer.

Objective To characterize the metabolic "fingerprint" of fecal extracts for diagnosis of early-stage colorectal cancer (CRC) using proton nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomics coupled with pattern recognition.Methods From January 2014 to December 2014,we collected fecal samples at the Second Affiliated Hospital of Shantou University Medical College,from 25 patients with colorectal adenomas (CR-Ad),20 with stage Ⅰ /Ⅱ CRC,and 32 healthy controls (HCs).The patients were diagnosed by histopathology.No subjects had any complicating diseases.HCs showed no abnormalities from blood tests,endoscopic examination,diagnostic imaging,and/or medical interviews.We excluded participants who used antibiotics,NSAIDS,statins,or probiotics within two months of study participation,and any patients who underwent chemotherapy or radiation treatments prior to surgery.We used orthogonal partial least-squares-discriminant analysis (OPLS-DA) for pattern recognition (dimension reduction) on 1H-NMR processed data (1H frequency of 400.13 MHz),to find metabolic differences among CR-Ad,carcinoma and HC fecal samples;and receiver operating characteristic (ROC) analysis to determine the diagnostic value of the fecal metabolic biomarkers.Results Fecal samples were collected from 20 patients with Stage Ⅰ/Ⅱ CRC (11 M,9 F,median age (52±13) years),25 with CR-Ad (14 M,11 F,median age (53±11) years) and 32 HCs (15 M,17 F,median age (53± 14) years).OPLS-DA clearly distinguished CR-Ad and stage Ⅰ/Ⅱ CRC from HC samples,based on their metabolomic profiles.Relative signal intensities in HCs were significantly lower than in the cancer patients for butyrate (HC:23.0±6.0;CR-Ad:18.0±5.0;CRC:14.0±6.0;Z=-2.07,P=0.008),acetate (HC:45.0± 11.0;CR-Ad:31.0±11.0;CRC:24.0±8.0;Z=-2.32,P=0.011),propionate (HC:26.0 ± 7.0;CR-Ad:22.0 ± 6.0;CRC:19.0 ± 5.0;Z=-2.43,P=0.032),glucose (HC:37.0±7.0;CR-Ad:31.0±7.0;CRC:26.0±8.0;Z=-2.07,P=0.044) and glutamine (HC:4.5±2.0;CR-Ad:4.9 ± 1.0;CRC:5.4 ± 1.0;Z=2.21,P=0.044).However,relative signal intensities in HCs were significantly higher than in patients for lactate (HC:4.8± 1.0;CR-Ad:6.9±2.0;CRC:4.8± 1.0;Z=2.02,P=0.038),glutamate (HC:3.2±2.0;CR-Ad:4.9 ± 1.0;CRC:3.2± 2.0;Z=2.21,P=0.044) and succinate (HC:12.0±2.0;CR-Ad:15.0±3.0;CRC:12.0± 2.0;Z=2.25,P=0.011).Among the potential biomarkers,acetate at 1.92 ppm,and succinate at 2.41 ppm displayed relatively high area under ROC,with sensitivity and specificity both ＞90％,to distinguish early-stage CRC patients from HCs.Conclusion Fecal metabolic profiles distinguish of HCs from patients with CRC patients,even in the early stages (stage Ⅰ/Ⅱ),highlighting the potential of NMR-based fecal metabolomic fingerprinting as tools for early CRC diagnosis.

Objective To characterize the metabolic "fingerprint" of fecal extracts for diagnosis of early-stage colorectal cancer (CRC) using proton nuclear magnetic resonance spectroscopy (1H-NMR)-based metabolomics coupled with pattern recognition.Methods From January 2014 to December 2014,we collected fecal samples at the Second Affiliated Hospital of Shantou University Medical College,from 25 patients with colorectal adenomas (CR-Ad),20 with stage Ⅰ /Ⅱ CRC,and 32 healthy controls (HCs).The patients were diagnosed by histopathology.No subjects had any complicating diseases.HCs showed no abnormalities from blood tests,endoscopic examination,diagnostic imaging,and/or medical interviews.We excluded participants who used antibiotics,NSAIDS,statins,or probiotics within two months of study participation,and any patients who underwent chemotherapy or radiation treatments prior to surgery.We used orthogonal partial least-squares-discriminant analysis (OPLS-DA) for pattern recognition (dimension reduction) on 1H-NMR processed data (1H frequency of 400.13 MHz),to find metabolic differences among CR-Ad,carcinoma and HC fecal samples;and receiver operating characteristic (ROC) analysis to determine the diagnostic value of the fecal metabolic biomarkers.Results Fecal samples were collected from 20 patients with Stage Ⅰ/Ⅱ CRC (11 M,9 F,median age (52±13) years),25 with CR-Ad (14 M,11 F,median age (53±11) years) and 32 HCs (15 M,17 F,median age (53± 14) years).OPLS-DA clearly distinguished CR-Ad and stage Ⅰ/Ⅱ CRC from HC samples,based on their metabolomic profiles.Relative signal intensities in HCs were significantly lower than in the cancer patients for butyrate (HC:23.0±6.0;CR-Ad:18.0±5.0;CRC:14.0±6.0;Z=-2.07,P=0.008),acetate (HC:45.0± 11.0;CR-Ad:31.0±11.0;CRC:24.0±8.0;Z=-2.32,P=0.011),propionate (HC:26.0 ± 7.0;CR-Ad:22.0 ± 6.0;CRC:19.0 ± 5.0;Z=-2.43,P=0.032),glucose (HC:37.0±7.0;CR-Ad:31.0±7.0;CRC:26.0±8.0;Z=-2.07,P=0.044) and glutamine (HC:4.5±2.0;CR-Ad:4.9 ± 1.0;CRC:5.4 ± 1.0;Z=2.21,P=0.044).However,relative signal intensities in HCs were significantly higher than in patients for lactate (HC:4.8± 1.0;CR-Ad:6.9±2.0;CRC:4.8± 1.0;Z=2.02,P=0.038),glutamate (HC:3.2±2.0;CR-Ad:4.9 ± 1.0;CRC:3.2± 2.0;Z=2.21,P=0.044) and succinate (HC:12.0±2.0;CR-Ad:15.0±3.0;CRC:12.0± 2.0;Z=2.25,P=0.011).Among the potential biomarkers,acetate at 1.92 ppm,and succinate at 2.41 ppm displayed relatively high area under ROC,with sensitivity and specificity both ＞90％,to distinguish early-stage CRC patients from HCs.Conclusion Fecal metabolic profiles distinguish of HCs from patients with CRC patients,even in the early stages (stage Ⅰ/Ⅱ),highlighting the potential of NMR-based fecal metabolomic fingerprinting as tools for early CRC diagnosis.

Objective: To explore the protective effect and mechanism of Somatostatin (SS) on the mice with Paraquat (PQ) poisoning, and to provide theoretical basis for clinical treatment of PQ poisoning. Methods: 48 SPF male BALB/c mice were randomly divided into control group, SS group, PQ group and PQ+SS group, with 12 mice in each group. 20 ml/kg SS solution was intraperitoneally injected into the SS group and PQ+SS group, and the same amount of normal saline was intraperitoneally injected into the PQ group and control group. After 1 hour of the above treatment, PQ group and PQ+SS group were given 60 mg/kg PQ solution by one-time gavage, while the control group and SS group were given the same amount of normal saline by gavage. After the above treatment for 3 hours, the SS group and PQ+SS group were intraperitoneally injected with SS solution (20 ml/kg) again, and the PQ group and the control group were intraperitoneally injected with the same amount of normal saline. 6 eyeballs were randomly selected from each group for blood collection, and the levels of TNF-α, MPO and il-6 in the blood of mice were detected by ELISA and other methods. The left lung was taken after blood collection to calculate the D/W ratio. The levels of SOD, caspase-3 and MDA were detected in some lung tissues by chemical colorimetry, and the amount of NF-κB was detected by Western blot. The lung histopathological changes were observed under light microscope. Results: The mice in the control group and SS group showed normal activity and good general condition; Mice in the PQ group ate less and moved less, responded slowly to stimulation, breathed shallow and fast with thickened breath sound, had messy and dull fur, and had varying degrees of cyanosis on their lips and limbs; The above performance of PQ+SS group was less than that of PQ group. Under the light microscope, the alveolar structure of PQ group was disordered and seriously damaged. The pathological changes of lung tissue in PQ+SS group were significantly improved compared with that in PQ group, and the pathological scores were decreased (all P<0.05) . Compared with the control group, there was no significant change in the levels of various indicators in the SS group (all P>0.05) . While the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 in PQ mice were significantly increased, and the levels of oxidative cytokines SOD and lung tissue D/W were significantly decreased (all P<0.05) . Compared with the PQ group, the PQ+SS group, the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 decreased, and the levels of oxidative cytokines SOD and lung tissue D/W increased (all P<0.05) . Conclusion PQ poisoning can cause lung injury in mice, while SS can alleviate lung injury in PQ poisoned mice, improve the general survival state of mice, and play a certain protective role in lung injury caused by PQ poisoning, which may be achieved by SS throμgh reducing the inflammatory response, oxidative stress response and lung tissue apoptosis in lung injury caused by PQ poisoning.