BACKGROUND:The development of vaccines is a promising and cost-effective strategy to prevent emerging multidrug-resistant (MDR) Acinetobacter baumannii (A. baumannii) infections. The purpose of this study was to prepare a multiepitope peptide nanovaccine and evaluate its immunogenicity and protective effect in BALB/c mice. METHODS:The B-cell and T-cell epitopes of Omp22 from A. baumannii were predicted using bioinformatics methods and identified by immunological experiments. The optimal epitopes were conjugated in series by 6-aminocaproic acid and chemically synthesized multiepitope polypeptide rOmp22. Then, rOmp22 was encapsulated by chitosan (CS) and poly (lactic-co-glycolic) acid (PLGA) to prepare CS-PLGA-rOmp22 nanoparticles (NPs). The immunogenicity and immunoprotective efficacy of the vaccine were evaluated in BALB/c mice. RESULTS:CS-PLGA-rOmp22 NPs were small (mean size of 272.83 nm) with apparently spherical structures, positively charged (4.39 mV) and nontoxic to A549 cells. A high encapsulation efficiency (54.94%) and a continuous slow release pattern were achieved. Compared with nonencapsulated rOmp22, CS-PLGA-rOmp22 immunized BALB/c mice induced higher levels of rOmp22-specific IgG in serum and IFN-γ in splenocyte supernatant. Additionally, lung injury and bacterial burdens in the lung and blood were suppressed, and potent protection (57.14%-83.3%) against acute lethal intratracheal A. baumannii challenge was observed in BALB/c mice vaccinated with CS-PLGA-rOmp22. CONCLUSION:CS-PLGA-rOmp22 NPs elicited specific IgG antibodies, Th1 cellular immunity and protection against acute lethal intratracheal A. baumannii challenge. Our results indicate that this nanovaccine is a desirable candidate for preventing A. baumannii infection.

Objective: To explore and analyze the risk factors of depression and anxiety in diabetics during perioperative period and study the nursing intervention.Methods: 101 patients with type-2 diabetes in perioperative period in our hospital were assessed by self-rating depression scale (SDS, n=61) and self-rating anxiety scale (SAS, n=40). The psychological factors and metabolic indexes were compared and analyzed. The patients with depression or anxiety were divided into special nursing group (n=30) and control group (n=30). Special nursing mainly included psychological comfort intervention, dietary nursing intervention and exercise guidance intervention. The effects of special nursing intervention on the improvement of depression or anxiety, metabolic indexes and quality of life were studied.Results: The psychological factors of patients with depression or anxiety were compared with those of patients without depression or anxiety during perioperative period of type-2 diabetes, including therapeutic effect, fear of disease and operation, medical atmosphere, worrying about carelessness of curing and nursing, pain degree, sleep quality, disease knowledge, marital relationship, family economic status, worrying about the influence of career, and metabolic indexes, including fasting blood glucose detection, 2-hour postprandial blood glucose and glycosylated hemoglobin, had significant differences (p<0.01). The Logistic multivariate analysis found that the above indicators were independent risk factors (p<0.05). After special nursing, the condition of depression and anxiety, metabolic indicators and qualityConclusion: The important factors of depression and anxiety in the perioperative period of type-2 diabetes included the above psychological factors and metabolic indicators. The use of special nursing intervention had a great assistant therapeutic effect on the condition of depression and anxiety of patients with type-2 diabetes mellitus in perioperative period.

Objective To analysed the clinical characteristics of the Carbapenems-resistant Pseudomonas aeruginosa (CRPA) infection.Methods 55 patients with CRPA infection and 85 patients with CSPA (the Carbapenems-sensitive Pseudomonas aeruginosa) infection were recruited in the First people's hospital of Kunshan city,from January to December 2017,and a retrospective cohort analysis was carried out.Results 32.72％ patients with CRPA infection were found in department of ICU,30.91％ in department of neurosurgery and 12.735 in department of neurology,significantly higher than the corresponding distribution of patients with CSPA infection,respectively 14.12％,8.24％ and 8.24％.In accord to this finding,most patients infected with CRPA suffered from existed brain trauma (41.82％) or stroke (30.91％).More fever (74.55％),shock (27.27％) and hypoproteinemia (70.91％) were found in CRPA group than in CSPA group (respectively 56.47％,15.29％ and 47.06％).It's indicated thatmultiple bacterial infection,mechanical ventilation and proton pump inhibitor were independent risk factors for CRPA infection.Conclusions Most of patients with CRPA infection in our hospital were seriously ill and likely to cause respiratory failure even multiple organ failure,leading to poor clinical prognosis.It's in urgent need to promote rational use of antibiotics in the early stage of the disease,strengthen the prevention of respiratory complications for patients withbrain trauma and cerebrovascular disease and improve the management of nosocomial infection for control of CRPA in hospital.

The intracellular survival of Mycobacterium tuberculosis (Mtb) has a central role in the pathogenesis of tuberculosis. Mtb Rv2346c is a member of 6-kDa early secreted antigenic target family of proteins, which are known to inhibit the host immune responses to promote bacillary persistence in macrophages. However, the mechanism through which Rv2346c participates in Mtb pathogenesis is unclear. In the present study, recombinant Rv2346c protein was synthesized and used to treat Bacillus Calmette–Guérin (BCG)-infected macrophages. The results showed that Rv2346c inhibited the proliferation of BCG-infected macrophages and enhanced the survival of BCG in macrophages. Tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were upregulated during BCG infection but downregulated by Rv2346c. Additional experiments showed that nuclear transcription factor-κB (NF-κB) in BCG-infected macrophages induced the production of TNF-α and IL-6. In addition, miR-155 and miR-99b had a suppressive effect on NF-κB, and the expression of these miRNAs was promoted by p38. Furthermore, Rv2346c was shown to decrease the activation of NF-κB, whereas it enhanced the phosphorylation of p38 and the expression of miR-155 and miR-99b. The function of Rv2346c was also verified in Mtb-infected mice. The results showed that Rv2346c increased the observed bacterial load and lung injury and downregulated TNF-α and IL-6 in vivo. Overall, our results reveal that Rv2346c enhances mycobacterial survival in macrophages via inhibiting the production of TNF-α and IL-6 in a p38/miRNA/NF-κB pathway-dependent manner, suggesting that Rv2346c acts as a crucial virulence factor in Mtb infection and has potential use as a target for anti-tuberculosis therapy.

Asthma is a chronic airway inflammatory disorder and progresses mainly due to airway remodeling. Chrysin, a natural flavonoid, has been reported to possess multiple biologic activities, including anti-inflammation, anti-oxidation and anti-proliferation. The present study aimed to investigate whether chrysin could relieve allergic airway inflammation and remodeling in a murine model of chronic asthma and the mechanism involved. The female BALB/c mice sensitized and challenged with ovalbumin (OVA) successfully developed airway hyperresponsiveness (AHR), inflammation and remodeling. The experimental data showed that chrysin could alleviate OVA-induced AHR. Chrysin could also reduce OVA-induced increases in the number of inflammatory cells, especially eosinophils, interleukin (IL) -4, and IL-13 in bronchoalveolar lavage fluid (BALF) and total IgE in serum. The decreased interferon-γ (IFN-γ) level in BALF was also upregulated by chrysin. In addition, inflammatory cell infiltration, goblet cell hyperplasia and the expression of α-smooth muscle actin (α-SMA) around bronchioles were suppressed by chrysin. Furthermore, the phosphorylation levels of Akt and extracellular signal-regulated kinase (ERK) could be decreased by chrysin, which are associated with airway smooth muscle cell (ASMC) proliferation. These results indicate the promising therapeutic effect of chrysin on chronic asthma, especially the progression of airway remodeling.

Although known for their broad spectrum and curative efficacy on drug-resistant pathogens and as nephrotoxicity-free, impairments were observed on renal function during clinical treatment of the two most commonly used fourth-generation cephalosporins: Cefpirome and cefepime. The present study aimed to further explore the exact influences of them on renal function. In vitro, the cell viability of renal cells cultured in drug-combined medium was tested for six dilutions. In vivo, a clinical cohort study was carried out to detect the influence of cefpirome and cefepime on the serum creatinine (SCr) level of patients. Cefpirome had an inhibition rate with half maximal inhibitory concentration (IC50) of 143.5 µmol/l on renal mesangial cells, which was greater compared to the IC50 of 7.702 µmol/l for cefepime. The clinical cohort study data revealed that cefpirome treatment could lead to a greater increase of the average SCr level compared to cefepime on days 3 and 7 during therapy, and in addition, a greater incidence of SCr >445 µmol/l, an indicator of clinical renal failure. Furthermore, patients with an average age >65 years were observed as more susceptible to an SCr rise caused by either cefpirome or cefepime, with a larger augment in the average SCr, as well as a higher incidence of SCr >445 µmol/l compared to patients aged <65 years. In conclusion, cefpirome may have more potential to cause renal impairment compared to cefepime, therefore, more caution and comprehensive analysis of patient conditions is required during the clinical choice of fourth-generation cephalosporins.

As a infectious disease which seriously harms human health,it is necessary to develop novel types of vaccine against tuberculosis.Breakthroughs in immunology,molecular genetics,genetic engineering,proteomics and other technologies have promoted the development of new tuberculosis vaccines.This paper elaborates the current status and prospects the development of these novel tuberculosis vaccines.

Infections with Pseudomonas aeruginosa have been a long-standing challenge for clinical therapy because of complex pathogenesis and resistance to antibiotics, thus attaching importance to explore effective vaccines for prevention and treatment. In the present study, we constructed a novel DNA vaccine by inserting mutated gene toxAm encoding Pseudomonas Exotoxin A and gene pcrV encoding tip protein of the type III secretion system into respective sites of a eukaryotic plasmid pIRES, named pIRES-toxAm-pcrV, and next evaluated the efficacy of the vaccine in murine acute Pseudomonas pneumonia models. Compared to DNA vaccines encoding single antigen, mice vaccinated with pIRES-toxAm-pcrV elicited higher levels of antigen-specific serum immunoglobulin G (IgG), enhanced splenic cell proliferation and cytokine secretion in response to Pseudomonas aeruginosa antigens, additionally PAO1 challenge in mice airway resulted in reduced bacteria burden and milder pathologic changes in lungs. Besides, it was observed that immunogenicity and protection could be promoted by the CpG ODN 1826 adjuvant. Taken together, it's revealed that recombinant DNA vaccine pIRES-toxAm-pcrV was a potential candidate for immunotherapy of Pseudomonas aeruginosa infection and the CpG ODN 1826 a potent stimulatory adjuvant for DNA vaccination.

Objective Exotoxin A ( encoded by gene toxA ) , one of the most toxic protein secreted by pseudomonas aerugi-nosa(P.a.), and PcrV (encoded by gene pcrV), key component to type Ⅲsecretion system of P.a., both matter significantly to the virulence of P.a.The article was to construct a novel DNA vaccine encoding a mutated toxA gene and the pcrV gene of P .a.and i-dentify gene expressions in eukaryotic cells . Methods The genes of toxA and pcrV were amplified by PCR , and the toxA gene was mutated to reduce the toxicity of Exotoxin A .Then gene fragments toxA m and pcrV were inserted into eukaryotic expression plasmid pIRES simultaneously to construct a recombinant DNA vaccine pIRES-toxAm-pcrV.The novel plasmid was transfected into HEK-293 cells by lipofectamine 2000 .The expressions of toxA m and pcrV were detected by Western blot . Results Gel electrophoresis demon-strated the target gene fragments encoding Exotoxin A and PcrV .Western blot exhibited proteins encoded toxA and pcrV expressed by HEK 293 cells. Conclusion The recombinant plasmid pIRES-toxAm-pcrV was successfully constructed .Western blot analysis indi-cated the expressions of toxA m and pcrV in HEK-293 cells.It may be used as a potential candidate of preventive vaccine of Pseudo-monas aeruginosa .

铜绿假单胞菌(Pseudomonas aeruginosa)为无荚膜、无芽孢、可运动的革兰阴性菌,专性需氧,广泛分布于水、土壤、空气、人和动物的皮肤和肠道中,在潮湿环境中能更好地生长.另外,医院的呼吸机、污水池、体温计、注射器和镊子等均可检出该菌.铜绿假单胞菌为条件致病菌,是医院感染的重要病原菌之一.目前铜绿假单胞菌的耐药情况日趋严重,多重耐药和广泛耐药使其临床治疗面临严峻挑战.急性铜绿假单胞菌感染若不能治愈而转为慢性后,其细菌表型会发生演变,形成生物膜,进一步降低抗菌药物的疗效.近年来,铜绿假单胞菌疫苗的研究为临床提供了新的治疗方向[1].传统疫苗制备工艺直接利用病原体作为免疫原,但病原体型别差异、减毒疫苗的潜在恢复感染危险和制备程序繁琐等不足限制了其大量应用.随着对铜绿假单胞菌致病机制的深入探讨和基因工程技术的发展,直接利用细菌的抗原表位基因序列构建核酸或核酸-载体疫苗已成为研究的主流方向[2-3].现就近年来铜绿假单胞菌的重组核酸疫苗的研究进行综述,在抗原表位选择、制备技术、免疫方案等方面进行初步探讨[3].