Role and pathogenic mechanism of TcpC in uropathogenic Escherichia coli-induced mouse model of cystitis / 中华微生物学和免疫学杂志

Objective:To investigate the role of TcpC in uropathogenic Escherichia coli (UPEC)-induced cystitis in mice and to preliminarily analyze the pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 CFU wild-type UPEC CFT073 (CFT073 wt) or tcpc gene-deleted mutant (CFT073 Δ tcpc) from urethra into bladder to construct the mouse model of cystitis. The mice were sacrificed 3 d after infection and the bladders were taken to observe the gross pathological changes. Histopathological changes in bladder tissues were observed after HE staining. Immunohistochemistry was used to detect TcpC in bladder tissues. Bacterial loads in urine samples of UPEC-infected mice were counted by tenfold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from the bladder and urine samples of CFT073 wt-infected mice was measured by PCR. Real-time quantitative RT-PCR (qRT-PCR) and Western blot were performed to detect the expression of TcpC at mRNA and protein levels in macrophages after CFT073 wt infection. The influence of UPEC strains on the activation of NF-κB signaling pathway in macrophages were determined by Western blot. The levels of proinflammatory factors and the bacterial and cell activity after infecting macrophages with UPEC strains were detected by ELISA, laser confocal microscope and fluorescence microscope, respectively. Results:Compared with the mice with CFT073 Δ tcpc infection, CFT073 wt-infected mice had significantly enlarged bladder and severe neutrophil infiltration and abundant TcpC in bladder tissues. The number of bacteria in the urine of CFT073 wt-infected mice was significantly greater than that of the CFT073 Δ tcpc group. PCR results showed that the bacteria in bladder or urine were CFT073 wt. The expression of TcpC at both mRNA and protein levels in macrophages increased significantly after CFT073 wt infection. Moreover, in CFT073 wt-infected macrophages, the expression of IκBα was promoted and the phosphorylation of p65 and the production of proinflammatory factors were suppressed. TcpC was instrumental in the survival and invasion of CFT073 wt in macrophages. Conclusions:TcpC expression increased significantly in mice with CFT073 wt-induced cystitis. TcpC inhibited the activation of NF-κB signaling pathway and the production of proinflammatory factors in macrophages to improve the survival rate of CFT073 wt, which was closely related to the pathogenesis and immune evasion of UPEC.

Hypervirulent Klebsiella pneumoniae serotype K1 induces liver abscess through activating NLRP3 inflammasome / 中华微生物学和免疫学杂志

Objective:To investigate whether the hypervirulent Klebsiella pneumoniae (hvKP) induces liver abscess through activating NLRP3 inflammasome. Methods:K1-hvKP and K35-non-hvKP bacterial suspensions were intraperitoneally injected into C57BL/6 mice to establish the models of liver abscess. Human peripheral blood neutrophils were sorted by immunomagnetic beads with CD45 + and Gr-1 + , and the purity was detected by flow cytometry. The concentrations of capsular polysaccharide of K1-hvKP and K35-non-hvKP were detected by total carbohydrate assay kit. The expression of IL-18 and IL-33 by neutrophils at mRNA and protein levels was detected by real-time fluorescence quantitative PCR and ELISA, respectively. The activation of NLRP3 inflammasome in neutrophils was detected by Western blot. Neutrophil extracellular trap formation (NETosis) was observed under confocal laser scanning microscope. Results:The C57BL/6 mice with K1-hvKP infection had significantly serious liver abscess as compared with the K35-non-hvKP-infected mice. The purity of human neutrophils was more than 95%. The concentration of capsular polysaccharide in K1-hvKP was significantly higher than that in K35-non-hvKP. Compared with K35-non-hvKP, K1-hvKP significantly promoted the neutrophils to express IL-18 and IL-33 at both mRNA and protein levels, enhanced the activation of NLRP3 and induced NETosis.Conclusions:This study suggested that hvKP could promote NETosis by activating NLRP3 inflammasome to cause liver abscess.

Signaling pathways involved in inhibition of macrophage phagocytosis by TcpC of uropathogenic Escherichia coli / 中华微生物学和免疫学杂志

Objective:To investigate the signaling pathway of inhibiting macrophage phagocytosis of TIR domain-containing protein encoded by Escherichia coli (TcpC) N-terminal ubiquitin ligase active fragments of uropathogenic Escherichia coli. Methods:Bioinformatics software was used to analyze the amino acid sequences and the function of TcpC N-terminal ubiquitin ligase active fragments as well as the functional sites. PCR was performed to amplify tcpc-330, tcpc-450 and tcpc-510 genes and a prokaryotic expression system was constructed to express the target proteins. The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were purified by Ni-NTA affinity chromatography. LPS in the recombinant proteins was removed by Detoxi-gel chromatography. The expression of MyD88 at protein and mRNA levels in macrophages incubated with rTcpC-N110, rTcpC-N150, rTcpC-N170 or rTcpC-TIR was detected by Western blot and qRT-PCR. The activation of NF-κB signal pathway and the levels of proinflammatory factors in macrophages incubated with the above TcpC protein fragments were measured by Western blot and ELISA, respectively. Results:Cys12, Trp104 and Trp106 in the N-terminal fragment of TcpC were crucial amino acids in maintaining its ubiquitin ligase activity. The target recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were successfully expressed and purified. After Detoxi-gel chromatography, rTcpC-N110, rTcpC-N150 and rTcpC-N170 extracts were undetectable for LPS. TcpC ubiquitin ligase fragments inhibited the expression of MyD88 at protein level, but not affect its expression at mRNA level in macrophages. LPS-induced phosphorylation of NF-κB signaling pathway-related proteins p50 and p65 was significantly inhibited in macrophages treated with TcpC ubiquitin ligase fragments. Moreover, LPS-induced production of pro-inflammatory factors was also significantly inhibited.Conclusions:The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 could inhibit the expression of MyD88 at protein level and suppress the activation of NF-κB signaling pathway, suggesting that they were closely related to the inhibition of innate immune activity of macrophages.

Transumbilical endoscopic surgery in the diagnosis of ascites of unknown origin / 中南大学学报(医学版)

To explore whether transumbilical endoscopic surgery (TUES) can effectively and safely elucidate the causes of ascites of unknown origin.
 Methods: A total of 23 consecutive patients with ascites of unknown origin who undertook TUES procedures in the Department of Gastroenterology of The Third Xiangya Hospital of Central South University between January 2014 and January 2016 were retrospectively investigated. Clinical manifestations, laboratory examinations and findings from radiological examinations and endoscopic investigations were noted before the procedure. Conditions of the abdominal cavity under endoscope, final diagnosis and outcome of patients were carefully recorded.
 Results: TUES procedure was successfully performed in all 23 patients with an operation time of (58.2±13.9) min. Twenty-two patients were undertaken biopsy on the nodules or masses that found in the abdominal cavity. Definite diagnoses were established in the overwhelming majority of patients (22/23). The most common causes of ascites for the 23 cases was tuberculosis (8 cases), followed by peritoneal carcinomatosis (6 cases), and pseudomyxoma peritonei (5 cases). Operation-related complications, such as postoperative bleeding, perforation, peritonitis, intraabdominal chronic abscesses, were not observed, except one case showed a transient moderate fever in 24 hours after operation. No mortality related to TUES occurred. We concluded that TUES was a feasible, economic and minimally invasive approach to access the peritoneal cavity.
 Conclusion: TUES combinated with biopsy can effectively elucidate the causes of ascites of unknown origin.

Role of the Wnt signaling pathway in the development and progression of liver fibrosis / 临床肝胆病杂志

Liver fibrosis has a complex pathogenesis and involves various intracellular and extracellular signal transduction pathways, but its specific pathogenesis remains unclear. Recent studies have confirmed that the activation of hepatic stellate cells due to abnormal activation of the Wnt signaling pathway is an important cause of liver fibrosis, and targeted inhibition of the Wnt signaling pathway can prevent the activation and proliferation of hepatic stellate cells and thus achieve an antifibrogenic effect. This article reviews the role of the Wnt signaling pathway in the development and progression of liver fibrosis and the latest research advances in the antifibrogenic effect of the interference of the Wnt signaling pathway, in order to provide new ideas for anti-hepatic fibrosis and the delay of early-stage cirrhosis.

Research advances in animal models of obstructive jaundice / 临床肝胆病杂志

Obstructive jaundice is a common disease that greatly threatens human health.In order to better investigate the pathogenesis of obstructive jaundice,pathophysiological changes during disease progression,and treatment measures,the establishment of a stable animal model of obstructive jaundice becomes the basis for the research on various types of obstructive jaundice.This article elaborates on the advantages and disadvantages of the animal models of acute,progressive,chronic fluctuant,malignant,and reducible obstructive jaundice,as well as their application in clinical practice.These animal models have certain limitations in reflecting the development and progression of target disease.Most animal models are established by surgery or external physical and chemical damage,and there are still no mouse models of gene knockout or overexpression.Future research will focus on the stable animal models of chronic and progressive obstructive jaundice and special types of obstructive jaundice.

Research advances in animal models of obstructive jaundice / 临床肝胆病杂志

Obstructive jaundice is a common disease that greatly threatens human health.In order to better investigate the pathogenesis of obstructive jaundice,pathophysiological changes during disease progression,and treatment measures,the establishment of a stable animal model of obstructive jaundice becomes the basis for the research on various types of obstructive jaundice.This article elaborates on the advantages and disadvantages of the animal models of acute,progressive,chronic fluctuant,malignant,and reducible obstructive jaundice,as well as their application in clinical practice.These animal models have certain limitations in reflecting the development and progression of target disease.Most animal models are established by surgery or external physical and chemical damage,and there are still no mouse models of gene knockout or overexpression.Future research will focus on the stable animal models of chronic and progressive obstructive jaundice and special types of obstructive jaundice.

Comparation of Simplified and Traditional Chinese Version of Psycho-educational Profile Applied in Special Children / 中国康复理论与实践

@#Objective To study the variability of Traditional Chinese version of the Psycho-educational Profile-3rd edition (PEP-3) and Simplified Chinese version of the Psycho-educational Profile (C-PEP) as evaluating the special children. Methods 194 special children were evaluated with C-PEP and PEP-3 from April, 2011 to December, 2014. The scores of cognitive verbal/preverbal, expressive language, recep-tive language, gross motor, fine motor, visual-motor imitation, and communication and physical ability were compared. Results There were significant diferences in all the dimensions between PEP-3 and C-PEP scales (Z>3.446, P<0.01) except cognitive verbal/preverbal (Z=0.912, P=0.362). Conclusion There was difference between PEP-3 and C-PEP for the evaluation of special children.

Application of iPad and its Software in Rehabilitation of Children with Autism:Four Cases Report / 中国康复理论与实践

Objective To explore the application of iPad-based software Let'S Talk in the rehabilitation of children with autism. Meth-ods From March to September, 2015, iPad-based software Let'S Talk was applied in four children with autism. The language ability, emo-tion and behavior control ability, self-consciousness and eye contact were observed. The Psycho-educational Profile-3rd (PEP-3), Autism Be-havior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were used to access the months of age, behavior characteristics, and the severity of autism before and after application. Results The language ability and behavior control ability improved, as well as the self-consciousness and eye contact, and the bad behaviors reduced. The months of age rose in PEP-3, and the scores of ABC and CARS de-creased. Conclusion IPad-based software Let'S Talk could be applied in the rehabilitation of children with autism.

Isocryptotanshinone Induced Apoptosis and Activated MAPK Signaling in Human Breast Cancer MCF-7 Cells / 한국유방암학회지

PURPOSE: Isocryptotanshinone (ICTS) is a natural bioactive product that is isolated from the roots of the widely used medical herb Salvia miltiorrhiza. However, few reports exist on the mechanisms underlying the therapeutic effects of ICTS. Here, we report that ICTS has anticancer activity and describe the mechanism underlying this effect. METHODS: The antiproliferative effect of ICTS was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and clonogenic assays. The effect of ICTS on the cell cycle was measured using flow cytometry. Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins. Finally, the effect of ICTS on mitogen-activated protein kinases (MAPKs) was determined by Western blotting. RESULTS: ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro. Among the tested cell lines, MCF-7 cells showed the highest sensitivity to ICTS. ICTS significantly inhibited colony formation by MCF-7 cells. Furthermore, exposure of MCF-7 cells to ICTS induced cell cycle arrest at the G1 phase and decreased mitochondrial membrane potential. Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells. In addition, ICTS activated MAPK signaling in MCF-7 cells by inducing time- and concentration-dependent phosphorylation of JNK, ERK, and p38 MAPK. CONCLUSION: Our results suggest that ICTS inhibited MCF-7 cell proliferation by inducing apoptosis and activating MAPK signaling pathways.