Diabetic kidney disease(DKD)is the leading cause of end-stage renal disease.Chinese materia medica shows unique advantages in intervening DKD.Silent information regulator 1(SIRT1)protein plays a vital role in the occurrence and development of DKD.This article was about the SIRT1 signaling pathway and the main molecular regulatory mechanisms to sort out the role of Chinese materia medica in regulating SIRT1 in podocytes,oxidative stress and inflammatory response.It was found that Chinese materia medica monomers and compounds were capable of regulating the PGC-1 α signaling pathway,Nrf2/ARE signaling pathway,AMPK signaling pathway,NF-κB p65 signaling pathway,and PI3K/AKT/FoxO1 signaling pathway by targeting SIRT1,exerting pharmacological effects such as inhibiting cell apoptosis,improving mitophagy,inhibiting inflammatory response,etc.,thus intervening in DKD,which can provide reference for relevant research.

Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K⁺ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β₂-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

Objective To validate whether the expression of human cluster of differentiation 55 (hCD55) protein in porcine islet cells could inhibit the activation of complement components in human serum. Methods Four adult pigs with WT (wild type), GTKO [α-1, 3-galactosyltransferase (GGTA1) knockout], GTKO/hCD55 and hCD55 genotypes were selected. Islet cells were isolated from WT, GTKO and GTKO/hCD55 pigs, and the purity and insulin secretion function were detected. The expression of hCD55 at the DNA, RNA and protein levels was analyzed by agarose gel electrophoresis, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry, respectively. Complement-dependent cytotoxicity assay and complement deposition assay were performed under the incubation conditions with fresh human serum. Results The purity of isolated porcine islet cells from three genotype pigs was > 75%, and the glycemic index was > 1. The expression of hCD55 messenger RNA(mRNA) and protein in GTKO/hCD55 porcine islet cells decreased the deposition of human complement component C3c and membrane-attacking complex C5b-9, and reduced the cytotoxicity. Conclusions The expression of hCD55 protein in porcine islet cells could inhibit the activation of human complement and reduce complement-mediated killing effect, indicating that hCD55 protein could exert complement protection effect on porcine islet cells. These findings provide theoretical basis for the application of hCD55 in islet xenotransplantation.

Objective:To investigate the efficacy of different doses of vitamin C (VC) in prevention and treatment of non-alcoholic fatty liver disease (NAFLD) in mice.Methods:C57BL/6 mice were fed with high-fat diet to establish NAFLD models. The experimental animals were divided into early prevention and later treatment groups. Both of these two experimental processes had five subgroups, including control, high-fat diet (HFD), low-dose vitamin C (LD-VC, 15 mg/kg per day), medium-dose vitamin C (MD-VC, 30 mg/kg per day) and high-dose vitamin C (HD-VC, 90 mg/kg per day) subgroup, with six mice in each subgroup. In the early prevention group, the mice were prophylactically received VC for 12 weeks. In the later treatment group, the mice were treated with different dose of VC for 12 weeks after fed with HFD for six weeks and confirmed NAFLD by liver pathology. The differences in body weight, perirenal adipose tissue mass and serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triacylglycerol (TG) were observed among different groups. The scores of hepatocyte steatosis, lobular inflammation and ballooning in liver histopathology of mice in each group were evaluated by non-alcoholic fatty liver disease activity score (NAS) scoring system. Tukey′s multiple comparison test and Kruskal-Wallis H test were performed for statistical analysis. Results:In the early prevention group, the body weight, perirenal adipose tissue mass, TG level and the score of liver steatosis of LD-VC subgroup were all lower than those of HFD subgroup ((30.27±0.94) g vs. (32.18±1.35) g, (0.25±0.05) g vs. (0.32±0.02) g, (0.25±0.02) mmol/L vs. (0.30±0.03) mmol/L, 0 vs. 1.0(1.0)). The body weight, perirenal adipose tissue mass, blood glucose level, TG level and score of liver steatosis of MD-VC subgroup were all lower than those of HFD subgroup ( (29.72±0.58) g vs. (32.18±1.35) g, (0.24±0.05) g vs. (0.32±0.02) g, (6.93±0.59) mmol/L vs. (8.33±1.02) mmol/L, (0.24±0.04) mmol/L vs. (0.30±0.03) mmol/L, 0 vs. 1.0(1.0)); meanwhile, the blood glucose level and TG level of HD-VC subgroup were both lower than those of HFD subgroup ((6.72±0.59) mmol/L vs. (8.33±1.02) mmol/L, (0.23±0.04) mmol/L vs. (0.30±0.03) mmol/L), and the differences were statistically significant (all P<0.05). In the later treatment group, TG level of LD-VC subgroup was lower than that of HFD subgroup ((0.25±0.07) mmol/L vs. (0.37±0.06) mmol/L); the body weight, perirenal adipose tissue mass, blood glucose level, TG level and liver steatosis score of MD-VC subgroup were lower than those of HFD subgroup ((29.93±1.28) g vs. (33.24±2.45) g, (0.29±0.08) g vs. (0.53±0.14) g, (7.63±0.57) mmol/L vs. (9.13±1.52) mmol/L, (0.23±0.03) mmol/L vs. (0.37±0.06) mmol/L, 0.5(1.0) vs. 2.0(1.0)); the blood glucose level and TG level of HD-VC subgroup were both lower than those of HFD subgroup ((7.20±0.72) mmol/L vs. (9.13±1.52) mmol/L, (0.19±0.03) mmol/L vs. (0.37±0.06) mmol/L); however the body weight, liver weight, perirenal adipose tissue mass and lobular inflammation score of HD-VC subgroup were all high than those of HFD subgroup( (36.34±2.44) g vs. (33.24±2.45) g, (1.18±0.07) g vs. (1.06±0.09) g, (0.78±0.17) g vs. (0.53±0.14) g, 1.0(1.0) vs.0(1.0)), and the differences were statistically significant (all P<0.05). The body weight, perirenal adipose tissue mass and the score of liver steatosis, lobular inflammation and ballooning of LD-VC subgroup of the early prevention group were all lower than those of LD-VC subgroup of the later treatment group ((30.27±0.94) g vs. (34.75±1.64) g, (0.25±0.05) g vs. (0.61±0.14) g, 0 vs.1.5(1.0), 0 vs. 0.5(1.0), 0 vs. 1.0(0)); and the body weight, liver weight, perirenal adipose tissue mass, ALT level, AST level and scores of liver steatosis and lobulor inflammation of HD-VC subgroup of the early prevention group were all lower than those of HD-VC subgroup of the late treatment group ((31.78±0.71) g vs. (36.34±2.44) g, (1.01±0.02) g vs. (1.18±0.07) g, (0.30±0.05) g vs. (0.78±0.17) g, (8.83±0.98) U/L vs. (12.75±2.05) U/L, (29.00±4.19) U/L vs. (41.88±14.36) U/L, 1.0(0) vs. 2.5(1.0), 0 vs. 1.0(1.0)), and the differences were statistically significant (all P<0.05). Conclusions:MD-VC can prevent the occurrence of NAFLD in mice at an early stage, and it is also benefit to the later treatment of NAFLD in mice. However, HD-VC has potential risks in early prevention and later treatment of NAFLD in mice.

Objective To explore the effect of platelet-rich plasma (PRP) on the generation of reactive oxygen species (ROS) and the phenotypes of photo-aging fibroblasts.Methods A photoaging cell model by repeating UVB irradiation was treated using appropriate concentration of PRP;Cell morphology and the rate of aging dying were observed under inverted microscope 24 hours later after establishment of the cell model;The expression of ROS between experimental and control group was detected using fluorescence microscope after single UVB irradiation.The relative intensity of fluorescence was analyzed using flow cytometry.Results PRP could ameliorate the large and sprawl appearance of photoaging fibroblasts obviously,reduce the generation of ROS as well as decrease the relative intensity of ROS.Conclusions PRP can decrease the level of intracellular oxidative stress caused by UVB irradiation,reduce the generation of ROS and ameliorate the senescence-like phenotypes of pho toaging fibroblasts.