The effects of Malassezia in the activation of Interleukin (IL)-23/IL-17 axis in Psoriasis.

We explore the association of Malassezia and IL-23/IL-17 axis in the skin lesions of patients with Psoriasis. From October 2018 to October 2020, 202 psoriasis patients were hospitalized in the dermatology department of Yantaishan hospital. The patients' skin lesions were collected, and Malassezia-specific mRNA in the skin lesions was determined. The patients were subdivided into Malassezia high and low distribution groups as per the Malassezia-specific mRNA results. Psoriasis Area and Severity Index (PASI) scores between the two groups were performed. LL-37, IL-23, IL-17A, and tumor necrosis factor α (TNF-α) expression in the skin lesions of the two groups were determined. Malassezia mRNA and the correlation of LL-37 with inflammatory factors TNF-α, IL-23, and IL-17A were determined. The relevance of inflammatory factors, Malassezia infection, and LL-37 content with PASI score were studied. The Malassezia high distribution group was treated with etoconazole, and the effects of treatment on the PASI score, IL-23, TNF-α, and IL-17A were determined. The PASI score, neutrophil, eosinophil, and peripheral blood white blood cell counts, and lgG in the Malassezia high distribution group were significantly higher than in the low distribution group (P<0.05). IL-23, LL-37, TNF-α, and IL-17A levels in the Malassezia high distribution group were significantly higher than in the low distribution group (P<0.05). Malassezia and LL-37 levels had a moderate positive correlation (R=0.5009, P<0.0001). Malassezia and LL-37, IL-17A, TNF-a, and IL- 23 correlated positively. Malassezia, IL-17A, LL37, TNF-a, and IL-23 correlated positively with the PASI score of Psoriasis. Ketoconazole therapy inhibited the PASI score, IL-23, TNF-a, and IL-17A expressions in patients. Malassezia enhances the progression of Psoriasis through the aberrant activation of the IL-23/IL-17 axis.

The Clinical Value of High-Quality Nursing in Concurrent Radiotherapy and Chemotherapy after Glioma Surgery and Its Influence on the Stress Indicators Cor, ACTH, and CRP.

OBJECTIVE: The purpose of this study is to explore the clinical value of high-quality nursing in concurrent radiotherapy and chemotherapy after glioma surgery and its influence on the stress indicators such as cortisol (Cor), adrenocorticotrophic hormone (ACTH), and C-reactive protein (CRP). METHODS: A total of 94 glioma patients diagnosed and treated in our hospital were randomly divided into a research group and a control group, with 47 cases in each group. Both groups of patients were given concurrent radiotherapy and chemotherapy. On this basis, patients in the control group were given basic care, while patients in the research group were given a combination of basic care and high-quality care. The nursing satisfaction and adverse reactions of the two groups were compared. The pain degree and the levels of stress indicators Cor, ACTH, and CRP at different time points were compared between the two groups. The sleep quality, bad mood, and quality of life before and after nursing were compared between the two groups. RESULTS: After nursing, the nursing satisfaction of the research group (95.74%) was higher than that of the control group (80.85%), and the difference between the two groups was statistically significant (X 2 = 11.678, P < 0.05). There was no significant difference between patients in the Visual Analogue Scale (VAS) score and the levels of stress indicators Cor, ACTH, and CRP at the T1 time point between the two groups (P > 0.05). With the passage of time, the levels of Cor and ACTH of the two groups showed an upward trend. At T4, the increased levels of Cor and ACTH in the research group were less than those in the control group, and the difference was statistically significant (P < 0.05). The VAS scores and CRP levels of the two groups showed an upward trend at T1 and T2 and a downward trend at T3 and T4. And, at T4, the decrease in CRP level of the research group was greater than that in the control group, and the difference was statistically significant (P < 0.05). Before nursing, there was no statistically significant difference between two groups of patients in the time to fall asleep, sleep time, number of awakenings, SAS score, self-rating depression scale (SDS) score, quality of life index scores, and total scores (P > 0.05). After nursing, the time to fall asleep and the number of awakenings in the two groups of patients showed an upward trend, and the increase in the control group was higher (P < 0.05). The sleep time of the two groups showed a downward trend, and the degree of decline in the control group was higher (P < 0.05). After nursing, the SAS score and SDS score of the two groups of patients decreased ( ∗ P < 0.05), and the decrease in the research group was more obvious (# P < 0.05). After nursing, the scores of all indicators of the quality of life and the total score of the two groups increased and the score of the research group increased more significantly (P < 0.05). After nursing, the control group had 5 cases of gastrointestinal reactions, 7 cases of bone marrow suppression, 6 cases of leukopenia, 6 cases of thrombocytopenia, and 10 cases of dizziness and nausea. In the research group, there were 1 case of gastrointestinal reaction, 2 cases of bone marrow suppression, 1 case of leukopenia, 1 case of thrombocytopenia, and 2 cases of dizziness and nausea. The difference between the two groups was statistically significant (P < 0.05). CONCLUSION: Glioma patients are given high-quality care during the course of concurrent radiotherapy and chemotherapy, which can reduce the pain and bad mood of the patient, reduce the stress response of the patient, and improve the quality of sleep and the quality of life of the patient, thereby improving nursing satisfaction and patients compliance, reducing adverse reactions, and having a good prognosis.

VP1 of Enterovirus 71 Protects Mice Against Enterovirus 71 and Coxsackievirus B3 in Lethal Challenge Experiment.

Enterovirus and Coxsackievirus are the major viruses that cause hand, foot, and mouth disease (HFMD) outbreaks worldwide. Several studies have shown the potential of viral envelope protein 1 (VP1) on providing protective effects from viral strains of different genotypes. However, whether VP1 has the cross-protection in Enteroviruses or Coxsackievirus has not been studied in-depth. In this study, the vp1 gene of Enterovirus 71 (EV71) and Coxsackievirus B3 (CB3) was inserted into the vector pET22b (+) to form the respective expression plasmids pEVP1 or pCVP1, and then transformed into Escherichia coli strain BL21 (DE3). The recombinant EVP1 or CVP1 protein was overexpressed successfully and effectively purified to homogeneity. Then, we identified that EVP1 and CVP1 protein could generate effectively specific humoral immunity and cellular immunity in mice, what's more, we determined the cross-protection of VP1 between EV71 and CB3 in a murine model. The results showed that immunization with EVP1 could effectively induce specific IgG and secretory IgA against CVP1 and the sera from EVP1-immunized mice could neutralize CB3 with mean titers 1:440. In contrast, no measurable neutralizing antibodies to EV71 were detected in CVP1-immunized mice. Then, newborn BALB/C mice, whose mother was immunized with EVP1 or CVP1, were administered with different lethal doses of EV71 or CB3. The EVP1 immunized group showed a 90% protective efficacy for a CB3 dosage of 120 LD50, but the CVP1 immunized group showed no significantly different protective efficacy against 15 LD50 of EV71 compared with the BSA immunized group. Hence, EVP1 is a promising subunit vaccine candidate against Enterovirus 71 and Coxsackievirus B3 caused HFMD.

Expression and Clinical Significance of miR-26a and Pleomorphic Adenoma Gene 1 (PLAG1) in Invasive Pituitary Adenoma.

BACKGROUND Although pituitary adenoma is a malignant tumor, it can present as invasive growth in some cases. MicroRNA (miR)-26a has been found to be abnormally highly expressed in pituitary adenoma, indicating possible involvement in pathogenesis. As a known target gene of miR-26a, PLAG1 has abnormally low expression in pituitary adenoma. The correlation between miR-26a or PLAG1 expressional abnormality and occurrence of pituitary adenoma is still unknown, as is its association with invasiveness of pituitary adenoma. MATERIAL AND METHODS Pituitary adenoma tissues, including both invasive and non-invasive subtypes, were collected from our Neurosurgery Department, in parallel with normal pituitary tissues from postmortem autopsy. qRT-PCR was used to detect mRNA expression of miR-26a and PLAG1, while Western blotting was used to test PLAG1 protein expression. The correlation between miR-26a and PLAG1, and with pathological features, were analyzed. ROC analysis revealed the utility of miR-26a and PLAG1 in differential diagnosis of invasive/non-invasive pituitary tumors and in analyzing their effects on patient prognosis. RESULTS MiR-26a was remarkably upregulated in pituitary tumors, while PLAG1 was downregulated, especially in invasive pituitary tumors. miR-26a and PLAG1 had higher diagnostic values for differentiating between invasive and non-invasive pituitary tumors (AUC=0.889 and 0.818, respectively). Those patients with miR-26 overexpression and PLAG1 downregulation had unfavorable prognosis. miR-26 and PLAG1 are independent factors affecting patient diagnosis. CONCLUSIONS MiR-26a can facilitate occurrence of pituitary tumor and invasiveness, probably via inhibiting PLAG1 expression.

N-terminus three residues deletion mutant of human beta-defensin 3 with remarkably enhanced salt-resistance.

In this study, we designed and synthesized three N-terminal deletion analogs of human beta-defensin 3 (hBD-3), namely, hBD-3Δ4, hBD-3Δ7, and hBD-3Δ10, to determine the effect of N-terminal residues on the antibacterial activity and salt resistance of these peptides. The antibacterial activities and salt resistance of hBD-3 and its analogs were tested against a broad range of standard and clinically isolated strains. The deletion of nine N-terminal residues significantly reduced the antibacterial activity of hBD-3 against most of tested strains, particularly Klebsiella pneumonia. Compared with hBD-3 and other analogs, the analog with a deletion of three residues, hBD-3Δ4, exhibited significantly higher antimicrobial activity against almost all the tested strains, especially Escherichia coli and Enterococcus faecium, at high NaCl concentrations. Given its broad spectrum of antimicrobial activity and high salt resistance, hBD-3Δ4 could serve as a promising template for new therapeutic antimicrobial agents.

Development of a fusion protein SNVP as substrate for assaying multi-serotype botulinum neurotoxins.

The SNARE super family has three core members, namely SNAP-25, VAMP-2, and syntaxin. SNAP-25 is cleaved by botulinum toxins (BoNTs)/A, /C, and /E, whereas VAMP-2 is the substrate for proteolytic BoNTs/B, /D, /F, and /G. In this study, we constructed a hybrid gene encoding the fusion protein SNVP that encompasses SNAP-25 residues Met1 to Gly206 and VAMP-2 residues Met1 to Lys94. The hybrid gene was cloned in a prokaryotic vector carrying an N-terminal pelB signal sequence and overexpressed in Escherichia coli BL21(DE3) Rosetta. To easily purify the protein, 6× His double-affinity tags were designed as the linker and C terminus of the fusion protein. SNVP was purified to homogeneity by affinity chromatography on a HisTrap FF column and determined to be more than 97% pure by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-terminal sequencing of the purified protein showed that signal peptide was successfully removed. The fusion protein SNVP contained the protease cleavage sites of all seven serotypes of BoNTs. SNVP was also proved to be recognized and cleaved by the endopeptidase of BoNTs (BoNT/A-LC, BoNT/B-LC, BoNT/E-LC, and BoNT/G-LC). The novel fusion substrate SNVP exhibited high biological activity under the optimal conditions, suggesting its potential use as a reagent for BoNT assay.

Immunogenicity of recombinant porcine attaching and effacing-associated protein compared with intimin fragment in Escherichia coli O157:H7-infected mice.

Enterohemorrhagic Escherichia coli (EHEC) causes a wide spectrum of food- and waterborne infectious diseases, including diarrhea, hemorrhagic colitis, and even hemolytic-uremic syndrome. Porcine attaching and effacing-associated protein (Paa) was first identified in a porcine enteropathogenic E. coli strain. It has been proven essential in the attaching and effacing mechanism of EHEC. However, the immunologic function of the Paa protein has yet to be established. In the present study, recombinant Paa protein was overexpressed successfully in engineered E. coli and effectively purified to homogeneity. Comparative experiments were carried out in mice with a known adhesion factor (intimin) as reference to investigate the immunogenicity of Paa. Intraperitoneal immunization of Paa protein in mice elicited significantly high levels of serum immunoglobulin G antibodies via Th2-mediated humoral immune response. In mice challenged with E. coli O157:H7, Paa protein exhibited immunological effectiveness against pathogenic bacteria colonization and excretion in vivo. Compared with the intimin, Paa showed better protective effect against E. coli O157:H7 infection in mice, particularly those challenged with high lethal doses of the pathogen. Seventy percent of the mice challenged with 50 minimal lethal dose (MLD) in the Paa group survived, whereas only 50% survived in the intimin group. This finding is the first description of the immunologic function of the Paa protein. These attributes provide support for the development of Paa-based vaccine, which can be beneficial in treating infectious diseases caused by E. coli O157:H7.

The effect of pH on growth of Clostridium botulinum type A and expression of bontA and botR during different growth stages.

In this study, the effects of pH on the growth, relative expressions of bontA and botR genes, and neurotoxin formation of foodborne pathogens Clostridium botulinum type A were systematically studied throughout its growth stage. As in the previous reports, no C. botulinum growth was observed at extremely acidic pH. However, the effect of alkaline pH on the growth and neurotoxin production of C. botulinum was first revealed in this study. The maximum growth rate at pH 9.0 was similar to that at other pH values, although the lag phase at pH 9.0 was 16 h longer than that at pH 8.0. The peak of bontA mRNA expression at pH 9.0 was only 15.5% compared with that at pH 7.0. However, the neurotoxin concentration quantified in the cultures did not differ significantly. BotR is a known regulatory protein of bontA. The quantitative relationship between bontA and botR at different growth stages was first determined in this study. The mRNA levels of bontA were found to be positively correlated with those of botR, and the ratio of the mRNA transcript varied with pH. All these findings provide important physiological information on C. botulinum and thereby contribute to the improvement of food safety.

Biochemical characteristics of New Delhi metallo-ß-lactamase-1 show unexpected difference to other MBLs.

New Delhi metallo-ß-lactamase (NDM-1) is a new metallo-ß-lactamase (MBL) that has recently emerged as a global threat because it confers bacteria with resistance to almost all clinically used ß-lactam antibiotics. To determine the molecular basis of this threat, NDM-1 was purified from Escherichia coli TransB (DE3) carrying cloned blaNDM-1 gene by an anion-exchange chromatography step followed by a gel permeation chromatography step. The purified enzyme was stable even in extremely alkaline buffer (pH 11) and reached its highest activity at a low temperature (15°C), which was different from other MBLs. The 50% inhibition concentration of EDTA against NDM-1 was 412 nM, which showed that NDM-1 was more susceptible to EDTA than other MBLs. The effects of zinc on NDM-1 differed between cephem and carbapenem complexes, but inhibition at high Zn(2+) concentration was observed for all of tested ß-lactam compounds.