Objective To evaluate the efficacy and safety of itraconazole injection in treatment of invasive fungal infection in the patients with multiple organ dysfunction syndrome.Methods Invasive fungal infection in 15 patients with multiple organ dysfunction syndrome was treated with 200 mg itraconazole injection,twice a day on the first two days,then once a day,for 8 to 14 days.During the treatment,the symptoms and signs,the results of chemical detection were recorded.Everyday fungus,sputum,stool smear or culture were performed.Results The recovery rate,effective rate,and fungal clearance in this trial with itraconazole injection were 80%(12/15),86.7%(13/15),and 86.7%(13/15) respectively.The side effect was not found.Conclusion Itraconazole injection is an effective and safe drug against invasive fungal infection in patients with multiple organ dysfunction syndrome.

Objective:To investigate the role of NOD-like receptor protein 3 (NLRP3) in the regulation of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected macrophages. Methods:Expression profiles of phagocytosis-related genes in PBS- and V. vulnificus-infected J774A.1 cells were analyzed by RNA-Seq. NLRP3-knockout (NLRP3 KO) J774A.1 cells were constructed using CRISPR-Cas9 gene-editing system. The phagocytosis of V. vulnificus and pHrodo RED-labelled Escherichia coli ( E. coli) bioparticles in parental and NLRP3 KO J774A.1 cells was detected by flow cytometry. Real-time PCR was performed to measure the expression of Fgr2 b gene at mRNA level in PBS- and V. vulnificus-treated parental and NLRP3 KO J774A.1 cells. Results:The expression of 18 phagocytosis-related genes was upregulated in V. vulnificus-infected J774A.1 cells than in PBS-treated J774A.1 cells ( P<0.05). There was a 5 bp deletion in the exon 2 of NLRP3 gene in NLRP3 KO J774A.1 cells, resulting in frameshift mutation and complete loss of NLRP3 expression. NLRP3 KO J774A.1 cells exhibited enhanced phagocytosis of V. vulnificus and pHrodo RED-labelled E. coli bioparticles than parental J774A.1 cells ( P<0.05). Besides, the expression of Fgr2 b gene at mRNA level was significantly increased in V. vulnificus-infected NLRP3 KO J774A.1 cells than in parental J774A.1 cells ( P<0.05). Conclusions:The phagocytosis of V. vulnificus in macrophages could be negatively regulated by NLRP3, which was possibly mediated through the regulation of Fgr2 b gene expression.

AIM To investigate the effect of recipient kidney function by CsA coadministration Ber used to induce immune tolerance in rats of allogenic cardiac transplantation. METHODS The authors established the SD to Wistar rats heterotopic cardiac transplantation model by Onos methods.Observe the cardiac allograft survival and levels of BUN and Cr in the recipients plasma. The recipients were classified into 5 groups randomly after heterotopic cardiac transplantation were performed. Group A (Wistar to Wistar)): Received placebo intraperitoneal injected for 21 days; Group B (SD to Wistar): Saline intraperitoneal injected for 21 days; Group C (SD to Wistar):CsA 2 mg?kg -1 ?d -1 intraperitoneal injected for 21 days; Group D(SD to Wistar):Ber 16 mg?kg -1 ?d -1 gastrointubation for 21 days; Group E(SD to Wistar): Ber 16 mg?kg -1 ?d -1 gastrointubation coadministration CsA 2 mg?kg -1 ?d -1 ip for 21 days. RESULTS The levels of BUN and Cr in recipint plasma is lower evidently compare with the group with CsA ip simply. CONCLUSION Ber can reduce the renal toxicity in recipients by CsA which was intraperitoneal injected (ip) over a long period time.

Objective:To investigate the cytotoxicity of a recombinant Vibrio vulnificus cytolysin membrane pore-forming peptide (rMpf) to J774A.1 cells as well as its influences on reactive oxygen species (ROS) and lysosomes, and to analyze the cellular localization of the rMpf. Methods:The rMpf was expressed using an Escherichia coli expression system and then used to treat murine J774A.1 cells in vitro. The viability of rMpf- and PBS-treated J774A.1 cells and the levels of ROS and lysosomes in these cells were analyzed by flow cytometry. Confocal microscopy was used to observe the cellular localization of rMpf in the J774A.1 cells. Results:No significant differences in cell viability, ROS production or lysosome formation in J774A.1 cells were found between low-dose (4 μg/ml) rMpf-treated and untreated groups. However, the cell viability, ROS production and lysosome formation were significantly decreased after treating J774A.1 cells with higher doses of rMpf (20 and 60 μg/ml). Moreover, the rMpf was found to localize in both the cytoplasm and nuclei of J774A.1 cells in a dose-dependent manner.Conclusions:The rMpf could localize in both the cytoplasm and nuclei of J774A.1 cells. It would inhibit the cellular ROS production and lysosome formation to damage macrophage function. This study provided a novel sight for understanding the cytotoxic domain and pathogenesis of Vibrio vulnificus cytolysin and other membrane pore-forming cytolysins.

Objective The relationship between toll like receptor 4 (TLR4) the single nucleotide polymor-phisms of gene (SNP) and ischemic stroke reperfusion injury was evaluated by meta-analysis ,which was de-signed to provide evidence-based medicine for the prevention of ischemic stroke reperfusion injury . Methods In Medline ,PubMed ,EMBASE ,Cochrane ,CBM ,Chinese Journal Net ,academic conference materials and dissertations ,we searched for comprehensive information on the relationship between TLR4 gene SNP (rs10759932 ,rs11536891 ,rs11536879) in ischemic stroke Cohort study and case-control literature to determine whether the gene SNP (rs10759932 ,rs11536891 ,rs11536879) was associated with ischemic stroke reperfusion injury by genotype comparison .The heterogeneity test was performed by Stata11 .0 .The heterogeneity test was used to calculate the OR value .The heterogeneity between different studies was analyzed quantitatively . The fixed effect model was used and the percentage I2 was calculated .Results Meta analysis showed that 1943 cases of ischemia-reperfusion injury and 5043 cases of control group were analyzed ,TLR4 gene SNP (rs10759932 ,rs11536891 ,rs11536879) was associated with the risk of ischemic stroke reperfusion injury ,the dominant fixed effect models were (OR=1 .653 ,95％ CI:1 .416 -1 .930 ;OR=1 .653 ,95％ CI:1 .416 -1 .930 ;OR=1 .653 ,95％ CI:1 .416-1 .930 ;);a co-dominant fixed effect model (OR=1 .525 ,95％ CI:1 .350 -1 .723 ;OR= 1 .653 ,95％ CI:1 .416 -1 .930 ;OR= 1 .653 ,95％ CI:1 .416 -1 .930) .Conclusion TLR4 gene SNP (rs10759932 ,rs11536891 ,rs11536879) was associated with the occurrence of ischemic stroke reperfusion inju-ry by Meta-analysis .

Objective:To investigate the effects of TNF-α knockout on liver and spleen neutrophil responses to Vibrio vulnificus bloodstream infection in a mouse model. Methods:(1) TNF-α-knockout (TNF-α -/-) and wild-type (WT) C57BL/6J mice aged 6-8 weeks were randomly divided into four groups with six in each group: uninfected WT group, infected WT group, uninfected TNF-α -/- group and infected TNF-α -/- group. The mouse model of bloodstream infection was constructed by intraperitoneal injection of Vibrio vulnificus CGMCC1.1758 (2×10 8 CFU/200 μl), while the mice in the uninfected groups were injected intraperitoneally with equal amount of PBS. (2) Liver immune cells and splenocytes were isolated 4 h after infection and subjected to analyze the percentages and numbers of neutrophils, and the changes in cell viability, cellular reactive oxygen species (ROS) level and phagocytosis by flow cytometry. In addition, effects of Vibrio vulnificus bloodstream infection on mTOR signaling pathway in murine neutrophils were evaluated in vivo. Results:(1)Compared with the uninfected WT group, the percentages and numbers of neutrophils in liver and spleen tissues of the infected WT group increased significantly. The percentage and number of liver neutrophils were significantly higher in the infected TNF-α -/- group than in the infected WT group, but no significant difference in spleen neutrophils was detected between the two groups. (2) Compared with the infected WT group, the phagocytosis of liver neutrophils rather than that of spleen neutrophils was enhanced in the infected TNF-α -/- group. (3) The survival rates of neutrophils in both liver and spleen were decreased, while the cellular ROS level was significantly increased in the infected WT group compared with those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had increased survival rates of both liver and spleen neutrophils, but decreased level of ROS. (4) The levels of p-AKT (S473) in liver and spleen neutrophils of the infected WT group were lower than those of the uninfected WT group. Compared with the infected WT group, the infected TNF-α -/- group had lower level of p-AKT (S473) in liver neutrophils, but higher p-AKT (S473) level in spleen neutrophils. There were no significant differences in p-4E-BP1(T37/46) levels between the uninfected WT group and the infected WT group. The p-4E-BP1 (T37/46) level in liver neutrophils was lower in the infected TNF-α -/- group than in the infected WT group, but no significant difference in p-4E-BP1 (T37/46) levels in spleen neutrophils was observed between the two groups. Conclusions:TNF-α had different effects on the neutrophils in spleen and liver tissues of mice with Vibrio vulnificus bloodstream infection. It played a critical role in regulating the recruitment, phagocytic function and mTOR signaling of liver neutrophils after Vibrio vulnificus infection in vivo.

LC3-associated phagocytosis (LAP) is a special phagocytosis occurring at the intersection of the two pathways of phagocytosis and autophagy. A hallmark event of the LAP process is the recruitment of microtubule-associated proteinⅠlight chain type 3-Ⅱ(LC3Ⅱ) to the phagosome surface of the monolayer membrane structure. The LAP pathway relies on the functions of the RUN domain and cysteine-rich domain containing, Beclin 1-interacting protein (Rubicon) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. The LC3-associated phagosome (LAPosome) binds to the lysosome to digest and degrade the contents. In recent years, increasing studies have found that LAP plays an important role in the infections caused by pathogenic microorganisms including fungi and bacteria. LAP is a crucial way in the host to resist and degrade the infection of pathogenic microorganisms. However, some pathogenic microorganisms can effectively escape from LAP in the host and even use LAPosome as a place for colonization and replication. This article summarized the recent progress in the role of LAP in host defense against pathogenic microorganism infection and the significance of it in the occurrence and development of diseases.

ObjectiveTo explore the molecular mechanism of Qidi Tangshen prescription （QDTS） in regulating podocyte pyroptosis in diabetes nephropathy （DN）. MethodThrough in vivo experiment， db/db mice were divided into the model group， QDTS group （3.34 g·kg^-1）， valsartan capsule group （10.29 mg·kg^-1）， with db/m mice serving as the normal control. Each group consisted of 8 mice， and they underwent continuous intervention for 8 weeks. After the last administration， mice were euthanized， and kidney pathological changes were observed. Additionally， the expression levels of pyroptosis-related indicators， including NOD-like receptor protein 3 （NLRP3）， Gasdermin D protein （GSDMD）， and interleukin-1β （IL-1β） protein， were examined. Through in vitro experiment， mouse podocytes were divided into the normal glucose group （5.5 mmol·L^-1 glucose）， high glucose group （35 mmol·L^-1 glucose）， DMSO group （35 mmol·L^-1 glucose+200 mg·L^-1 DMSO）， and QDTS group （35 mmol·L^-1 glucose+200 mg·L^-1 QDTS freeze-dried powder）. After 48 hours of intervention， the expression levels of NLRP3， GSDMD， and IL-1β proteins were measured in podocytes. A drug-ingredient-target-disease interaction network for QDTS in the treatment of DN was constructed by network pharmacology methods. The key signaling pathways regulating podocyte pyroptosis were analyzed， and validation was conducted through in vivo and in vitro experiments. ResultCompared with normal group， glomerular hyperplasia and glomerular basement membrane thickening were observed in model group， and some segments were accompanied by obvious podocellular process fusion. The protein expressions of NLRP3， GSDMD and IL-1β in mouse kidney were increased， the protein expressions of mitogen-activated protein kinase 14 （MAPK14）， V-Rel reticuloendotheliosis virus oncogene homology A （RELA） and Caspase-8 in mouse kidney were increased （P<0.05）. Compared with model group， kidney pathological injury of mice in QDTS group was significantly reduced， and the expressions of NLRP3， GSDMD and IL-1β in kidney of mice in QDTS group and valsartan group were decreased （P<0.05）. The protein expressions of MAPK14， RELA and Caspase-8 in kidney of mice in QDTS group and valsartan group were decreased （P<0.05）. Network pharmacology results showed that there were 16 targets for QDTS to regulate DN cell pyrodeath， among which MAPK14， RELA and Caspase-8 were the key targets. Compared with normal glucose group， the protein expressions of NLRP3， GSDMD and IL-1β in high glucose group were increased （P<0.05）， and the protein expressions of MAPK14， RELA and Caspase-8 in mouse podocytes were increased （P<0.05）. Compared with high glucose group， the expressions of NLRP3， GSDMD and IL-1β in podocytes of mice in QDTS group were decreased （P<0.05）， and the expressions of MAPK14， RELA and Caspase-8 in podocytes of mice in QDTS group were decreased （P<0.05）. ConclusionQDTS reduces damage to DN podocytes， which is associated with its regulation of the MAPK14/RELA/Caspase-8 signaling pathway and inhibition of podocyte pyroptosis.

ObjectiveTo explore the mechanism of Qidi Tangshen prescription （QDTS） in alleviating podocyte injury and reducing urinary protein in diabetic nephropathy （DN）. MethodUsing network pharmacology methods， we collected the chemical components and targets of QDTS， as well as the targets related to DN. Subsequently， we constructed a "drug-ingredient-target-disease" network for QDTS in the treatment of DN to systematically elucidate the mechanism. The db/db mice were assigned into the model， QDTS （3.34 g·kg^-1）， and losartan capsules （10.29 mg·kg^-1） groups， and db/m mice served as the normal group. Each group consisted of 8 mice， and they underwent continuous intervention for 8 weeks. After the last administration， mice were euthanized， and the urinary albumin excretion rate （UAER） and renal pathological changes were measured and observed. The expression levels of protein kinase B1 （Akt1）， hypoxia-inducible factor-1 alpha （HIF-1α）， phosphorylated B-cell lymphoma-extra-large （p-Bcl-xl）， as well as autophagy-related indicators microtubule-associated protein 1 light chain 3 （LC3）， ubiquitin-binding protein p62 （p62）， and autophagy-related gene 6 homolog （Beclin1）， were determined. Furthermore， mouse podocytes were divided into the normal glucose （5.5 mmol·L^-1）， high glucose （35 mmol·L^-1）， DMSO （35 mmol·L^-1 glucose+200 mg·L^-1 DMSO）， and QDTS （35 mmol·L^-1 glucose+200 mg·L^-1 QDTS freeze-dried powder） groups. After 48 h of intervention， the protein levels of Akt1， HIF-1α， p-Bcl-xl， LC3， p62， and Beclin1 in podocytes were measured. ResultQDTS had 34 active components acting on 143 targets in the treatment of DN， and 55 targets were related to autophagy， in which Akt1， HIF-1α， and Bcl-xl were the key targets. Compared with the normal group， mice in the model group exhibited significantly increased UAER， glomerular hypertrophy， deposition of blue collagen fibers， thickening of the glomerular basement membrane， and noticeable fusion of podocyte foot processes in some segments. Furthermore， the modeling up-regulated the protein levels of p-Akt1， HIF-1α， and p62 and down-regulating the protein levels of p-Bcl-xl， LC3， and Beclin1 in the renal tissue （P<0.05）. Compared with the model group， QDTS and losartan decreased UAER （P<0.05） and alleviated the pathological damage in the renal tissue. Moreover， QDTS and losartan down-regulated the protein levels of p-Akt1， HIF-1α， and p62 and up-regulated the protein levels of p-Bcl-xl， LC3， and Beclin1 in the renal tissue （P<0.05）. In comparison to the normal glucose group， the high glucose group displayed up-regulated protein levels of p-Akt1， HIF-1α， and p62 and down-regulated protein levels of p-Bcl-xl， LC3， and Beclin1 in podocytes （P<0.05）. Compared with the high glucose group， QDTS down-regulated the protein levels of p-Akt1， HIF-1α， and p62 and up-regulated the protein levels of p-Bcl-xl， LC3， and Beclin1 in podocytes （P<0.05）. ConclusionQDTS alleviates podocyte damage and reduced urinary protein in DN by regulating the Akt1/HIF-1α/Bcl-xl signaling pathway， thereby enhancing podocyte autophagy.