Diabetic nephropathy (DN), as one of the most common complications of diabetes, is a primary cause of end-stage renal disease. The pathogenesis of DN encompasses processes such as chronic inflammation, recruitment and activation of immune cells, tubular and glomerular injury, and renal fibrosis. These processes are highly correlated with the activation of the nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 (NLRP3) inflammasome and the resulting pyroptosis it mediates. Previous studies have shown that the release of pro-inflammatory cytokines, leakage of damage-associated molecular patterns (DAMPs), recruitment and activation of immune cells can be reduced by regulating the NLRP3 inflammasome and its mediated pyroptosis, thereby slowing the diffusion of inflammatory responses in adjacentcells, fibrosis, and tissue remodeling processes. Ultimately, these process can improve renal injury and dysfunction caused by diabetic nephropathy. This article summarizes the molecular regulatory mechanisms of the NLRP3 inflammasome and its mediated pyroptosis at different pathological stages of DN, proposes potential targets for regulating their activation, aiming to provide a new direction for personalized treatment of DN.

Objective:To evaluate the role of O-linked-β-N-acetylglucosamine (O-GlcNAc) transferase (OGT) in methane-induced alleviation of oxygen-glucose deprivation and restoration (OGD/R) injury to rat spinal cord neurons and the relationship with Nrf2 expression.Methods:The primarily cultured spinal cord neurons of rats were seeded in 6-well plates at a density of 1×10 5 cells/ml and divided into 6 groups ( n=60 each) using a random number table method: control group (group C), group OGD/R, methane group (group M), and methane plus OGT inhibitor alloxan group (group MA). The medium was replaced with glucose- and serum-free Earle′s salt solution, and the neurons were exposed to 37 ℃ and 5%CO 2-95%N 2 in an incubator for 2 h followed by routine culture to establish the model of OGD/R.In group M, 200 μl methane-saturated saline (final concentration of methane 1.8 mmol/L) was added at oxygen-glucose restoration.Alloxan 8 mmol/L was added at 10 min after oxygen-glucose restoration to inhibit OGT in MA group.At 12 h of oxygen-glucose restoration, the neuronal survival rate, leakage rate of lactic dehydrogenase (LDH) and apoptotic rate of neurons were measured.The expression of OGT and H3K4me3 in Nrf2 promoter was measured by chromatin immunoprecipitation and fluorescent quantitative real-time polymerase chain reaction.Nucleoprotein was extracted for determination of O-GlcNAc glycosylation of RBBP5 (a H3K4 methyltransferase subunit) by immunoprecipitation and Western blot.The spatial proximity of MLL1 subunit of MLL complex to histone H3 was detected by proximity ligation assay.The expression of Nrf2 protein and mRNA was detected by Western blot and quantitative real-time polymerase chain reaction, respectively.The activities of superoxide dismutase (SOD) and catalase (CAT) and malonaldehyde (MDA) concentration were measured by enzyme-linked immunosorbent assay. Results:Compared with group C, the survival rate of neurons was significantly decreased, and the leakage rate of LDH, apoptotic rate and MDA content in supernatant were increased in OGD/R and M groups ( P<0.01). Compared with OGD/R, the survival rate of neurons was significantly increased, the leakage rate of LDH, apoptotic rate and MDA content in supernatant were decreased, the expression of OGT and H3K4me3 in Nrf2 promoter was up-regulated, O-GlcNAc glycosylation and spatial proximity level of MLL1 to histone H3 were increased, the expression of Nrf2 protein and mRNA was up-regulated, and the activities of SOD and CAT were increased in group M ( P<0.01). Compared with group M, the survival rate of neurons was significantly decreased, the leakage rate of LDH, apoptotic rate and MDA concentration were increased, the expression of OGT and H3K4me3 in Nrf2 promoter was down-regulated, O-GlcNAc glycosylation and spatial proximity level of MLL1 to histone H3 were decreased, the expression of Nrf2 protein and mRNA was down-regulated, and activities of SOD and CAT were decreased in group MA ( P<0.05 or 0.01). Conclusion:OGT is involved in methane-induced alleviation of OGD/R injury to rat spinal cord neurons, which is related to up-regulating Nrf2 expression.

Objective:To evaluate the role of reactive oxygen species (ROS) /nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in desflurane preconditioning-induced reduction of lipopolysaccharide (LPS)-induced acute cerebral inflammation in rats.Methods:One hundred and twenty clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-250 g, were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), LPS-induced acute cerebral inflammation group (group L), desflurane preconditioning group (group D), and N-acetylcysteine plus desflurane preconditioning group (group ND). LPS 1 mg/kg was injected via the tail vein to establish the model of acute cerebral inflammation in L, D and ND groups.In group D, rats inhaled 8.2% desflurane for 1 h once a day for 5 consecutive days, and LPS was intravenously injected at 24 h after the end of the 5th inhalation.In group ND, ROS scavenger N-acetylcysteine 150 mg/kg was intraperitonealy injected at 30 min before each inhalation of desflurane, and the other treatments were similar to those previously described in group D. The expression of Nrf2 and kelch-like ECH-associated protein 1 (Keap1) in microglia and astrocytes in the hippocampal CA1 region was determined by immunofluorescence double staining before LPS injection (at 24 h after the last desflurane preconditioning). Morris water maze test was used to measure the escape latency, space exploration time spent in the original platform quadrant, and frequency of crossing the original platform at 6, 12 and 24 h after injecting LPS.The expression of NOD-like receptor family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, interleukin-18 (IL-18) and interleukin-1beta (IL-1β) in microglia and astrocytes in the hippocampal CA1 region was determined by immunofluorescence double staining at 6, 12 and 24 h after injecting LPS.Nissl staining and immunofluorescence were used to count normal neurons and activated microglia and astrocytes in the hippocampus at 24 h after LPS injection. Results:Compared with group C, no significant change was found in the expression of Nrf2 and Keap1 in microglia and astrocytes before LPS injection ( P>0.05), and the escape latency was significantly prolonged, the space exploration time spent in the original platform quadrant was shortened, the frequency of crossing the original platform was decreased, the number of normal neurons was decreased, the number of activated microglia and astrocytes was increased, and the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was up-regulated after injecting LPS in group L ( P<0.05). Compared with group L, the expression of Nrf2 was significantly up-regulated, and the expression of Keap1 was down-regulated before injecting LPS ( P<0.05), and the escape latency was shortened, the space exploration time spent in the original platform quadrant was prolonged, the frequency of crossing the original platform was increased, the number of normal neurons was increased, the number of activated microglia and astrocytes was reduced, and the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was down-regulated after injecting LPS in group D( P<0.05). Compared with group D, the expression of Nrf2 was significantly down-regulated, the expression of Keap1 was up-regulated before injecting LPS ( P<0.05), and the escape latency was significantly prolonged, the space exploration time spent in the original platform quadrant was shortened, the frequency of crossing the original platform was decreased, the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was up-regulated, the number of normal neurons was reduced, and the number of activated microglia and astrocytes was increased after LPS injection in group ND ( P<0.05). Conclusion:ROS/Nrf2 signaling pathway is involved in desflurane preconditioning-induced reduction of LPS-induced acute cerebral inflammation in rats.

Objective:To analyze the epidemiological and molecular characteristics of human brucellosis in Qinghai province from 2005 to 2019 and provide basic data for brucellosis prevention and control.Method:The data about human brucellosis in Qinghai from 2005 to 2019 were collected from the information system of China CDC to describe the spatial, population and time distributions of human brucellosis cases in Qinghai. The isolated strains were identified and typed with traditional methods, BCSP31-PCR, AMOS-PCR and multi-locus variablenumber tandem repeat (MLVA-16).Results:A total of 577 human brucellosis cases were reported in Qinghai from 2005 to 2019, the average prevalence rate was 0.07 per 100 000 person, there were statistic differences among different years. The disease occurred all the year around, but mainly during March-October. The 577 cases were distributed in 31 counties (cities/districts) from 6 autonomous prefectures (cities). The prevalence rats of five counties were high, i.e. Menyuan Hui autonomous county (22.88 %, 132/577), Tianjun county (10.57 %, 61/577)、Xining city (10.57 %, 61/577), Henan Mongol Autonomous County (10.51 %, 58/577) and Haiyan county (9.53 %, 55/577). Age of the cases ranged from 8 years to 82 years, and the male to female ratio of the cases was 1.8∶1 (374/203). The prevalence rate in herdsman (47.83 %, 276/577) was highest among different occupational populations. Ten isolates were all Brucella melitensis strains, belonging to biovar 3, and clustering analysis indicated that the 10 strains had 5 genotypes, in which 2 were distinct, the remaining 3 were same. MLVA-16 analysis indicated that the 10 strains had close relationship with 26 B. melitensis strains isolated in Qinghai previously. Conclusions:The prevalence of brucellosis increased in Qinghai in recent years, we should strengthen the population based brucellosis surveillance and reporting. MLVA-16 indicated the gene diversity of the Brucella strains, suggesting that MLVA-16 can be used for genetic diversity analysis and molecular epidemiology survey to improve brucellosis surveillance.

Objective To investigate the diagnostic value of serum human epididymis protein 4 (HE4) ,carbohydrate antigen 125 (CA125) ,carbohydrate antigen antigen (CA199) ,carbohydrate antigen 153 (CA153) and alpha fetoprotein (AFP) in the early di-agnosis of ovarian cancer .Methods From February 2014 to October 2016 ,117 patients with ovarian cancers who were treated in this hospital were selected ,including 69 cases of ovarian cancer and 48 cases of benign ovarian lesions ,and 70 healthy volunteers were selected as control group .The serum levels of HE4 ,CA125 ,CA199 ,CA153 and AFP were measured in all subjects . Results The positive rates of HE4 ,CA125 ,CA199 ,CA153 and AFP in the ovarian cancer group were 59 .42% ,68 .12% ,33 .33% , 46 .38% and 39 .13% ,respectively ,which were significantly higher than those in the benign ovarian lesion group and the control group (P<0 .05) .The sensitivity of CA125 in the diagnosis of ovarian cancer was 68 .11% ,the specificity was 88 .98% ,the nega-tive predictive value was 78 .33% ,the positive predictive value was 82 .68% ,Youden index was 0 .571 .The diagnostic efficiency was better than that of other tumor markers .Pathological examination revealed 34 cases of serous adenocarcinoma ,18 cases of mucinous adenocarcinoma and 17 cases of endometrioid carcinoma in 69 cases of ovarian cancer .The positive rate of serous adenocarcinoma CA125 was 85 .29% ,significantly higher than mucinous carcinoma and endometrioid carcinoma (χ2 =9 .398 ,P<0 .05) .Conclusion CA125 has a good application value in the early diagnosis of ovarian cancer ,the positive rate is higher in serous adenocarcinoma .

Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.

Objective To obtain the prevalence data on human T Lymphotropic virus (HTLV) infection in 2 339 blood donors of an endemic coastal region of Fujian, China. Methods Serum antibodies to HTLV in the donors were detected by a home made double antigen sandwitch ELISA kit. All the ELISA positive samples were further confirmed by Western blot (WB) and/or polymerase chain reaction (PCR) combined with sequencing. Results Nine samples were confirmed as HTLV 1 infection among the 2 339 donors. A man whose wife is one of the nine was also detected to infect by WB and PCR. Conclusion The prevalence of HTLV infection was 0.38% in the endemic coastal region of Fujian, China.

Objective It was reported that pretreatment with small dose of lipopolysaccharide (LPS) could protect the animal from lethal dose endotoxin. The purpose of this study was to investigate the effects of pretreatment with small dose of LPS on cardiac function in endotoxemic rats and the possible mechanism. Methods Sixty male Wistar rats weighing 200-280 g were randomly divided into 3 groups: group I normal saline(NS) ( n = 12); group Ⅱ LPS (n = 24) and group Ⅲ LPS-pretreatment ( n = 24). Group Ⅱ and group Ⅲ were further divided into 3 groups: 2 h,4 h and 6 h subgroups based the time when blood sample was taken and the animal was sacrificed. The table shows the LPS given in the 3 groups:groupⅠⅡ Ⅲ0hNSNSLPS 0.25 mg?kg-1ip24 hNSNSLPS0.5mg?kg-1ip96 hNSLPS 10 mg?kg-1 Ⅳ LPS 10 mg?kg-1 TVThe animals were anesthetized with 3 % pentobarbital 30 mg?kg ip and intubated. Right femoral artery and vein were cannulated for MAP monitoring and fluid infusion. Cardiac catheter was placed in the left ventricle for measurement of left ventricular systolic pressure (LVSP) and?dp/dt max. Blood samples were taken 2 h,4 h and 6 h after intravenous LPS (10 mg?kg-1) for determination of plasma levels of L-lactate-dehydrogenase (LDH) and creatine kinase (CK) . The animals were sacrificed right after blood sampling and the heart was removed for determination of myocardial HSP 70 expression using immuno-histochemical staining. Results LVSP and?dp/dt max gradually decreased 1h after intravenous administration of LPS in group Ⅱ (LPS group); while in group DI (pretreatment group) the cardiac contractility was maintained and LVSP,?dp/dt max did not decrease as compared with the baseline value. Plasma LDH concentration and CK activity increased significantly 4 h and 6 h after intravenous IPS in group Ⅱ . The plasma LDH and CK levels were significantly lower in groupⅢthan those in group Ⅱ ( P

Objective: To observe the effect of Ginkgo Biloba extract (EGb) on Form deprivation Myopia (FDM) in the chick. Methods: 30 two day chicks were divided into three groups (Control Group, Drug Group Ⅰ, Drug Group Ⅱ). All of them were monocularly deprived by suturation of eyelids. Axial length was measured by A Scan ultrasonograph, the posterior cartilaginous sclera, the sclera fibrous and retina were observed under the light microscope. Results: Both refraction and axial eye length were changed by form deprivation in control group. refraction change ( P