Objective:Clamping bilateral renal arteries with refined surgical methods to establish the rat renal ischemia-reperfusion injury (RIRI) model, and study the protective mechanism of ischemic preconditioning renal (IPC) tubular cell-derived exosomes in RIRI.Methods:25 female Sprague Dawley (SD) rats were divided into sham group, model group, inactivated group, normoxic group, IPC group. In the sham operation group, after bilateral renal arteries were dissociated, the back incision was disinfected and closed. The model group established RIRI model; RIRI models were established in inactivated group, normoxia group and IPC group, and then 200 μg of inactivated exosomes, normal exosomes and IPC exosomes were injected into the caudal vein 24 hours after operation. Serum creatinine (Scr) and urea nitrogen (BUN) levels were detected. The pathological changes of renal tissue were observed under light microscope. Transmission electron microscopy (TEM) was used to observe the shape and size of renal tubular exosomes. Nanoparticle tracking analysis (NTA)was used to detect the concentration and size of renal tubular exosomes.Results:Compared with the sham group, the Scr and BUN levels in the model group were significantly elevated ( P<0.01). Renal pathological changes in the model group showed damaged of the tubular structure, necrosis and shedding of tubular epithelial cells, and a large number of inflammatory cells accumulated in the renal interstitial tissue with varying degrees of edema. Compared with the inactivated group, the Scr and BUN levels significantly decreased in the normoxic group and IPC group ( P<0.01). Renal pathological changes in the normoxic group and IPC group showed that the renal tubular cell necrosis alleviated, inflammatory was reduced, the improved edema. Compared with the normoxic group, the Scr and BUN levels in the IPC group were further reduced ( P<0.01). Renal pathological changes in the IPC group showed that the inflammatory cells were significantly reduced, the cell edema was significantly improved, and the cell apoptosis was significantly reduced. Conclusions:Clamping bilateral renal arteries with refined surgical methods is the main and optimal way to build a rat model of RIRI. IPC tubular cell-derived exosomes have protective and repair effects on RIRI.

Objective:To establish a mouse model of intra-jugular arteriovenous fistula (AVF) to screen differentially expressed genes in the process of intimal stenosis of AVF for investigating the abnormal expression signaling pathways and the mechanisms.Methods:Forty-six male C57BL/6 mice were randomly divided into AVF group ( n=23) and sham-operated group ( n=23). The AVF group underwent internal jugular arteriovenous fistuloplasty, and the sham-operated group separated the right external jugular vein and common carotid artery and then sutured the incision. The whole-genome sequences of mice with AVF stenosis were determined by transcriptomic reversible chain terminator and synthetic sequencing. The microarray data set was established, and the Benjamini & Hochberg method of gene microarray data analysis was applied to screen the differentially expressed genes. The differentially expressed genes were screened by R-language enrichment analysis. Then, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were performed. The subcellular localization of the differentially expressed genes was performed by BUSCA software. The protein network interaction of differentially expressed genes was analyzed by using STRING database and Cytoscape software. Results:In the AVF group, 21 mice were successfully modeled and 2 mice failed. Therefore, there were 21 mice in the AVF group and only 21 mice in the sham-operated group. This mouse internal jugular AVF model was innovated using the continuous-interrupted suture method, which improved the success rate of modeling this model. The differential gene sequencing analysis showed that there were 2 514 differentially expressed genes in the AVF process, including 1 323 up-regulated genes and 1 191 down-regulated genes. GO functional enrichment analysis showed that the differential genes were mainly enriched in metabolic process, activation, redox, mitochondria and so on. KEGG pathway enrichment analysis showed that the differential genes were enriched in metabolism, energy substance synthesis, diabetes, oxidative stress and so on. Statistical analysis of subcellular localization showed that the differences were mainly in mitochondrial proteins (24.24%), cytoplasmic proteins (17.51%), nuclear proteins (13.13%), cell membrane proteins (11.45%), and extracellular proteins (10.77%).Conclusions:Mitochondrial oxidative stress injury may be involved in the pathological damage process of endothelial proliferation stenosis in the AVF.

【Objective】 To investigate and analyze the clinical blood use and full-time professional technical personnel of Blood Transfusion Departments in tertiary hospitals in the Yangtze River Delta, in order to provide basis for health administration and medical institutions to make decisions on clinical blood transfusion management and standardize the construction of Blood Transfusion Departments. 【Methods】 According to the Clinical Blood Quality Control Index (2019 Edition), 435 tertiary hospitals in the Yangtze River Delta were selected for statistical analysis. 【Results】 Among the 435 tertiary hospitals, 316 (72.64%, 316/435) set up independent Blood Transfusion Departments, but 119 (27.36%, 119/435) did not. 295 (67.82%, 295/435) met the national standards for Blood Transfusion Department, and 263 (60.46%, 263/435) met the local standards. Only 5 of the top 10 tertiary hospitals with the most consumption of blood in the Yangtze River Delta met the national standards for the allocation of blood transfusion professional and technical personnel, and 17 hospitals met the local standards. Nine of the top 10 tertiary hospitals with the most beds met the national standards for blood transfusion professional and technical personnel, and 19 met the local standards. Among the full-time professional technicians in Blood Transfusion Department, medical technicians, physicians and nurses accounted for 91.03%, 6.46% and 2.52%; doctor, master, bachelor and below accounted for 1.74%, 12.4% and 85.86%; senior, intermediate, junior and below titles accounted for 18.82%, 38.61% and 42.58%, respectively. 【Conclusion】 Standardized construction of Blood Transfusion Department of tertiary hospitals in the Yangtze River Delta, including organization and talent team construction, needs to be strengthened to improve scientific and rational use of clinical blood.

Objective:To investigate the effects and underlying mechanisms of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/NF-κB signaling pathway in human kidney-2(HK-2) cells of hyperuricemic nephropathy.Methods:HK-2 cells were cultured in vitro and randomly divided into control group and experimental group. The experimental group was induced by high uric acid (720 μmol/L) immersion for 48 h to establish a cell model of hyperuricemic nephropathy in vitro and subsequently divided into hyperuricemic group, overexpressed protease activated receptor 2 (PAR2) and knockdown PAR2 group. The expressions of PAR2, PI3K, AKT, NF-κB mRNA were measured by real-time PCR. The expressions of PAR2, PI3K, AKT and NF-κB protein were measured by Western blotting. The expressions of tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), pro-interleukin-1β (pro-IL-1β), interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) were detected by enzyme linked immunosorbent assay (ELISA). Results:(1) Compared with the control group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in hyperuricemic group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant in hyperuricemic group were significantly increased (all P<0.01). (2) Compared with the hyperuricemic group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in overexpressed PAR2 group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, IL-1β and TGF-β1 in the supernatant were significantly increased (all P<0.05). (3) Compared with the hyperuricemic group, the expression of PAR2, PI3K, AKT and NF-κB mRNA and protein in knockdown PAR2 group were significantly decreased (all P<0.05), the expressions of IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant were significantly decreased (all P<0.05). Conclusions:In the process of uric acid-induced HK-2 cell damage, uric acid significantly up-regulates the expression of PI3K/AKT/NF-κB signaling pathway by activating PAR2, leading to a marked increase in inflammatory damage. Knocking down PAR2 inhibits the expression of PI3K/AKT/NF-κB signaling pathway, which can effectively reduce the inflammatory damage of HK-2 cells.

Objective To investigate the effect of β-lactam antibiotics on the false positive rate of the serum Aspergillus galactomannan (GM) assay in patients with lung diseases.Methods We selectively recruited 77 lung disease patients who did not meet the diagnostic criteria of invasive pulmonary Aspergillosis (IPA) and received different β-lactam antibiotics,while 41 patients without IPA who did not receive any antibiotic treatment were recruited as the control group.Serum samples for GM detection were collected from all participants.The rate of false-positive Aspergillus galactomannan was compared between the two groups.Results False-positive serum results were found in patients who received piperacillin-tazobactam (30.8％ or 8/26) and cefoperazone sulbactamand (27.8％ or 5/18).The rate of false-positive Aspergillus galactomannan in patients who receive β-lactam antibiotics were significantly higher than that in the control group (24.7％ or 19/77vs.7.3％ or 3/41,x2 =5.315,P=0.025).Taking false-positive serum Aspergillus galactomannan as the dependent variable and β-lactam antibiotic treatment as the independent variable,univariate logistic regression analysis showed that the rate of false-positive Aspergillus galactomannan in patients who received β-lactam antibiotics were 4.149 times more than that in the control group (OR=4.149,P=0.030).Conclusions The administration of β-lactam antibiotics may increase the occurrence of false-positive serum Aspergillus galactomannan,and physicians should be aware of this possible interference.

OBJECTIVETo explore the practicability of microskin grafting by spraying in burn management.

METHODSRazor thin autologous skin from pigs or burn patients was harvested and cut to pieces of 0.2 - 0.5 mm in size and suspended in normal saline. The suspension was put into a bottle with outlet and pumping device. The microskin suspended in the saline was sprayed to the burn wound and/or onto the alloskin sheets. The microskin distribution was detected by digital image analysis technique. In animal experiments, the burn wound development and pathomorphological changes after operation were observed. In burn patients who would receive microskin grafting, spraying method was used with the traditional flotation method as control. The treatment results and the operational procedures were compared between these two kinds of operation styles.

RESULTSThe microskin dispersion degree with spraying was much smaller than that with flotating method. In animal experiment with spraying method, the wound healing time was 23.2, 24.5 and 38.3 days in 3 groups, respectively, with the area ratio of donor to wound of 110, 120 and 150. In clinical study, The average operating time was 133.3 min with spraying method and 165.6 min with flotation method respectively (P < 0.05). The area ratio of donor to wound was 118.8 with spraying and 17.6 with flotation methods, respectively (P < 0.01). The one time wound coverage rate was 92.6% with spraying and 79.7% with flotation methods (P < 0.01). The wound healing time was 29.7 days with spraying and 37.3 days with flotating methods, respectively (P < 0.05).

CONCLUSIONSpraying method of microskin grafting might be a good method in major burn treatment. The advantages with this method included well-distributed microskin, simpler handling, saving of donor skin, shortening of operating time and less time needed for the wound healing. It might be recommended for other wound covering materials.

Animals ; Burns ; surgery ; therapy ; Culture Techniques ; Dermatologic Surgical Procedures ; Female ; Humans ; Male ; Rabbits ; Skin ; injuries ; Skin Transplantation ; methods ; Transplantation, Heterologous ; Wound Healing