Baicalin and probenecid protect against Glaesserella parasuis challenge in a piglet model.

Glaesserella parasuis (G. parasuis) induces vascular damage and systemic inflammation. However, the mechanism by which it causes vascular damage is currently unclear. Baicalin has important anti-inflammatory, antibacterial and immunomodulatory functions. In this study, we explored the ability of baicalin and probenecid to protect against G. parasuis challenge in a piglet model. Sixty piglets were randomly divided into a control group; an infection group; a probenecid group; and 25 mg/kg, 50 mg/kg and 100 mg/kg baicalin groups. The probenecid group and the 25 mg/kg, 50 mg/kg and 100 mg/kg baicalin groups were injected intramuscularly with 20 mg/kg body weight (BW) probenecid and 25 mg/kg BW, 50 mg/kg BW and 100 mg/kg BW baicalin, respectively. All piglets except those from the control group were injected intraperitoneally with 1 × 108 CFU of G. parasuis. The control group was injected intraperitoneally with TSB. The results showed baicalin and probenecid protected piglets against G. parasuis challenge, improved body weight and decreased temperature changes in piglets. Baicalin and probenecid attenuated IL-1ß, IL-10, IL-18, TNF-α and IFN-γ mRNA levels in the blood for 48 h, inhibited the production of the nucleosides ATP, ADP, AMP and UMP from 24 to 72 h, reduced Panx-1/P2Y6/P2X7 expression, weakened NF-kB, AP-1, NLRP3/Caspase-1 and ROCK/MLCK/MLC signalling activation, and upregulated VE-cadherin expression in the blood vessels of piglets challenged with G. parasuis. Baicalin and probenecid alleviated pathological tissue damage in piglets induced by G. parasuis. Our results might provide a promising strategy to control and treat G. parasuis infection in the clinical setting.

Baicalin attenuates PD-1/PD-L1 axis-induced immunosuppression in piglets challenged with Glaesserella parasuis by inhibiting the PI3K/Akt/mTOR and RAS/MEK/ERK signalling pathways.

Infection of piglets with Glaesserella parasuis (G. parasuis) induces host immunosuppression. However, the mechanism underlying the immunosuppression of piglets remains unclear. Activation of the PD-1/PD-L1 axis has been shown to trigger host immunosuppression. Baicalin possesses anti-inflammatory and immunomodulatory functions. However, whether baicalin inhibits PD-1/PD-L1 activation and thus alleviates host immunosuppression has not been investigated. In this study, the effect of baicalin on the attenuation of piglet immunosuppression induced by G. parasuis was evaluated. Seventy piglets were randomly divided into the control group, infection group, levamisole group, BMS-1 group, 25 mg/kg baicalin group, 50 mg/kg baicalin group and 100 mg/kg baicalin group. Following pretreatment with levamisole, BMS-1 or baicalin, the piglets were challenged with 1 × 108 CFU of G. parasuis. Our results showed that baicalin, levamisole and BMS-1 modified routine blood indicators and biochemical parameters; downregulated IL-1ß, IL-10, IL-18, TNF-α and IFN-γ mRNA expression; and upregulated IL-2 and IL-8 mRNA expression in blood. Baicalin, levamisole and BMS-1 increased the proportions of CD3+ T cells, CD3+CD4+ T cells, CD3+CD8+ T cells and CD3-CD21+ B cells in the splenocyte population, increased the proportions of CD3+ T cells, CD3+CD4+ T cells and CD3+CD8+ T cells in the blood, and inhibited PD-1/PD-L1 and TIM-3 activation. Baicalin, levamisole and BMS-1 reduced p-PI3K, p-Akt, and p-mTOR expression, the p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 ratios and increased RAS expression. Baicalin, levamisole and BMS-1 provided substantial protection against G. parasuis challenge and relieved tissue histopathological damage. Our findings might provide new strategies for controlling G. parasuis infection and other immunosuppressive diseases.

PD-1/PD-L1 axis induced host immunosuppression via PI3K/Akt/mTOR signalling pathway in piglets infected by Glaesserella Parasuis.

Glaesserella parasuis, an important respiratory bacterial pathogen, causes Glässer's disease in piglets, with potential immunosuppression. We established a piglet infection model and explored the immunosuppression mechanism to improve our understanding of the host immune response to G. parasuis. Twenty piglets were randomly divided into two groups (n = 10). The infection group was intraperitoneally challenged with 2 × 108 CFU of G. parasuis in 2 mL TSB. The control group was intraperitoneally injected with equivalent TSB. After 72 h, the piglets were sacrificed, and spleen tissue was collected. PD-1/PD-L1 expression was determined. The splenocytes were isolated to detect CD3+ T, CD3+CD4+ T, CD3+CD8+ T and CD3-CD21+cell differentiation. Via data-independent acquisition (DIA), we compared the proteomics of healthy and infected spleen tissues. Glaesserella parasuis modified CD3+ T, CD3+CD4+ T, CD3+CD8+ T and CD3-CD21+ cell differentiation and PD-1/PD-L1 expression in the spleen. The infection group had 596 proteins with significant differences in expression, of which 301 were significantly upregulated and 295 downregulated. Differentially expressed proteins (DEPs) were mainly related to immune responses. This is the first study on PD-1/PD-L1 expression in the spleen associated with immunosuppression in a piglet model to explore the protein changes related to immune responses via DIA.

Widespread of non-typeable Haemophilus influenzae with high genetic diversity after two decades use of Hib vaccine in China.

BACKGROUND: The aim of this study was to analyze the microbiological characteristics of nasopharyngeal carriage Haemophilus influenzae isolates collected from children with respiratory infections in Beijing hospital and Youyang Hospital of China. METHODS: The serotypes of all isolates were determined using latex agglutinated antisera (a-f). The minimum inhibitory concentrations (MICs) of 11 antibiotics were determined using E-test strips. For the beta-lactamase-negative ampicillin-resistant (BLNAR) isolates, ftsI gene was sequenced based on fragments amplified by PCR. STs of H influenzae isolates were determined by multi-locus sequence typing. RESULTS: The overall carriage rate of H influenzae in the study population was 9.1% (362/3984). One hundred and ninety H influenzae isolates which were selected in our study were non-typeable (NTHi) and 44 (23.2%) of them were positive for ß-lactamase. All isolates were susceptible to ceftriaxone and levofloxacin. Susceptibility rates to erythromycin and sulfamethoxazole-trimethoprim in Beijing were significantly higher than Youyang (P < .05). Thirty-six BLNAR isolates were identified. The MLST analysis showed 108 STs in 190 isolates, the most common of which were ST408 (11, 5.8%), ST914 (10, 5.3%), ST57 (9, 4.7%), and ST834 (6, 3.2%). Twelve STs were detected in both of the study sites, which covered 63 isolates. CONCLUSIONS: All isolates in the present study were NTHi, which suggested widespread of this type in China. The BLNAR isolates were detected more frequently than before. Because high genetic diversity of NTHi isolates of H influenzae exists worldwide, it is important to continuously monitor these bacteria in the future.

Modified projection algorithms for solving the split equality problems.

The split equality problem (SEP) has extraordinary utility and broad applicability in many areas of applied mathematics. Recently, Byrne and Moudafi (2013) proposed a CQ algorithm for solving it. In this paper, we propose a modification for the CQ algorithm, which computes the stepsize adaptively and performs an additional projection step onto two half-spaces in each iteration. We further propose a relaxation scheme for the self-adaptive projection algorithm by using projections onto half-spaces instead of those onto the original convex sets, which is much more practical. Weak convergence results for both algorithms are analyzed.

Anemoside B4 attenuates necrotic enteritis of laying hens induced by Clostridium perfringens via inhibiting NF-κB and PI3K/Akt/mTOR signalling pathways.

Poultry necrotic enteritis is an important enteric disease which might be controlled by antibiotics. However, with the excessive use of antibiotics, the phenomenon of drug resistance of Clostridium perfringens is becoming increasingly prominent. Anemoside B4 exhibits important anti-inflammatory, antioxidant and immunomodulatory effects. This study was performed to estimate the effect of Anemoside B4 on chicken necrotic enteritis induced by C. perfringens in vivo and in vitro. In the in vivo experiment we investigated the efficacy of Anemoside B4 on the growth curve, biofilm formation, haemolytic activity, virulence-related gene expression and NF-κB and PI3K/AKT/mTOR activation in Caco-2 cells induced by C. perfringens. The results showed that 12.5-50 µg/mL Anemoside B4 had no antibacterial activity but could inhibit biofilm formation, attenuate haemolytic activity and virulence-related gene expression of C. perfringens and weaken NF-κB and PI3K/Akt/mTOR activation triggered by C. perfringens in Caco-2 cells. In the in vivo experiment, 60 17-day-old healthy White Leghorns were randomly divided into six groups. The growing laying hens of the control group were fed a basic diet, and those of the five challenged groups were fed a basic diet (infection group), added 0.43 g/kg Anemoside B4 (0.43 g/kg Ane group), 0.86 g/kg Anemoside B4 (0.86 g/kg Ane group), 1.72 g/kg Anemoside B4 (1.72 g/kg Ane group) and 40 mg/kg lincomycin (lincomycin group), respectively. All challenged laying hens were infected with 1 × 109 CFU C. perfringens from day 17-20. Blood and intestinal samples were obtained, and the data demonstrated that Anemoside B4 improved the blood biochemical parameters, attenuated jejunum tissue injury, increased the spleen, thymus, bursa of fabricius index, and decreased lesion scores of the jejunum and the ileum. In the jejunum, Anemoside B4 and lincomycin downregulated the expression of IL-1ß, IL-6, IL-10, TNF-α and IFN-γ at mRNA levels. Moreover, Anemoside B4 significantly enhanced both mRNA and protein levels of tight junctions ZO-1, Claudin-1 and MUC-2 in the jejunum. Anemoside B4 weakened p-P65, p-PI3K, p-Akt and p-mTOR protein expression in the jejunum infected by C. perfringens. Diets supplemented with Anemoside B4 alleviated C. perfringens-induced necrotic enteritis in laying hens by inhibiting NF-κB and PI3K/Akt/mTOR signalling pathways and improving intestinal barrier functions.

Serotype distribution and antimicrobial resistance patterns of invasive pneumococcal disease isolates from children in mainland China-a systematic review.

OBJECTIVES: This study aimed to review and report the serotype distribution and antimicrobial resistance patterns of invasive pneumococcal disease (IPD) isolates, as this information is important for policy making since China has not adopted any pneumococcal vaccines in the national immunization schedule. METHODS: A systematic review of the published literature from January 2000 to December 2018 was performed to identify articles that describe the serotype and/or antimicrobial resistance patterns of IPD cases in children in mainland China. Analysis of the extracted data was performed with the Microsoft Excel spreadsheet program. The percentage of the serotypes was calculated by dividing the number of isolates for each serotype with the total number of isolates included in all the studies. The theoretical impact of the vaccine was estimated by calculating the percentage of isolates that exhibited the serotypes included in the vaccines. The prevalence of antimicrobial resistance was defined as the number of isolates that were resistant divided by the total number of isolates tested for resistance to the specific antimicrobial agent. RESULTS: Forty-two articles were screened in the preliminary search, of which sixteen fulfilled inclusion criteria and were included in the final analysis. The predominant serotypes were 19A, 19F, 14, 23F, and 6B, and the estimated impact of PCV13 was 90.4%. The isolates exhibited a high frequency of resistance to cefuroxime, cefaclor, and erythromycin. CONCLUSIONS: It is necessary for Chinese children to receive PCV13. Clinical workers should pay attention to the high frequency of resistance to antimicrobial agents.

Simultaneous and semi-alternating projection algorithms for solving split equality problems.

In this article, we first introduce two simultaneous projection algorithms for solving the split equality problem by using a new choice of the stepsize, and then propose two semi-alternating projection algorithms. The weak convergence of the proposed algorithms is analyzed under standard conditions. As applications, we extend the results to solve the split feasibility problem. Finally, a numerical example is presented to illustrate the efficiency and advantage of the proposed algorithms.

An existence-uniqueness theorem and alternating contraction projection methods for inverse variational inequalities.

Let C be a nonempty closed convex subset of a real Hilbert space H with inner product 〈 ⋅ , ⋅ 〉 , and let f : H → H be a nonlinear operator. Consider the inverse variational inequality (in short, IVI ( C , f ) ) problem of finding a point ξ ∗ ∈ H such that f ( ξ ∗ ) ∈ C , 〈 ξ ∗ , v - f ( ξ ∗ ) 〉 ≥ 0 , ∀ v ∈ C . In this paper, we prove that IVI ( C , f ) has a unique solution if f is Lipschitz continuous and strongly monotone, which essentially improves the relevant result in (Luo and Yang in Optim. Lett. 8:1261-1272, 2014). Based on this result, an iterative algorithm, named the alternating contraction projection method (ACPM), is proposed for solving Lipschitz continuous and strongly monotone inverse variational inequalities. The strong convergence of the ACPM is proved and the convergence rate estimate is obtained. Furthermore, for the case that the structure of C is very complex and the projection operator P C is difficult to calculate, we introduce the alternating contraction relaxation projection method (ACRPM) and prove its strong convergence. Some numerical experiments are provided to show the practicability and effectiveness of our algorithms. Our results in this paper extend and improve the related existing results.

Henoch-Schönlein purpura associated with a neuroblastoma: Report of one case and a review of the literature.

Malignancies such as solid tumors and hematologic malignancies can often induce or be associated with Henoch-Schönlein purpura (HSP) in older males but not in children. Described here is the case of a 5-year-old boy who clinically presented with HSP. An imaging study of the abdomen revealed a right retroperitoneal neoplasm that histopathology postoperatively confirmed to be a neuroblastoma. Malignancies are sometimes associated with HSP mostly in older males, though children are affected, albeit rarely. Thus, all patients with HSP must be carefully examined to identify or exclude an underlying disease.

Molecular epidemic features and variation of rotavirus among children with diarrhea in Lanzhou, China, 2001-2006.

BACKGROUND: Human rotavirus (HRV) is the most common pathogen causing severe diarrhea among infants and young children worldwide. This study aims to understand rotavirus epidemiology and its variation in the period of 2001-2006 in Lanzhou, Gansu Province, China, and to provide an epidemiological basis for the development of rotavirus vaccine. METHODS: A total of 1019 stool specimens were collected from patients with acute diarrhea admitted to the First Hospital of Lanzhou University from 2001 to 2006, who were younger than 5 years old. Dako IDEIATM kits were used for detection of rotavirus, and RT-PCR was performed for determination of G serotype and P genotype of the rotavirus. RESULTS: Rotavirus was present in 51.6% (526) of the 1019 specimens. G serotype identified G3 at 40.9%, G2 14.6%, G1 22.2% and G9 1.9%. Mixed-G infection was observed in 4.4% and non-typeable infection 16.0%. P genotype was observed in 372 samples, of which P[8] accounted for 186 cases (50.0%), P[4] 72 cases (19.4%), mixed-P infection 2 cases (0.5%), and non-typeable cases 112 (30.1%). G3 was the most prevalent G serotype found in this study from 2001 to 2004, G2 was the most prevalent G serotype (34.4%) from 2004 to 2005, and G1 (61.5%) was the most prevalent strain from 2005 to 2006. G9 was detected in 10 cases (1.9%) and G4 was not detected during this 5-year period. P[8] was the most prevalent P genotype found over the 5 consecutive years of this study, although there was a significant transition of P genotype from 2004 to 2005 with P[4] (45%) identified as the predominant P genotype, followed by P[8] (22.1%). The predominant G-P combination was P[8]G1 (33.6%), followed by P[8]G3 (32.1%) and P[4]G2 (17.2%). Rotavirus diarrhea admissions peaked between October and December. Continuous surveillance showed that the incidence rate of rotavirus was the highest in infants aged 6-23 months, averaging 11.0-11.9 months. CONCLUSIONS: Five years of continuous surveillance showed that rotavirus remains the most significant viral agent causing diarrhea hospitalization among children under 5 years old in Lanzhou, China although the predominant strain of rotavirus varies between years. Mixed-G serotype infection also appears to occur at a relatively high rate in Lanzhou.

[Molecular epidemiology of human caliciviruses diarrhea among infants and young children in Lanzhou from December 2001 to June 2004].

OBJECTIVE: To investigate the characteristics of human caliciviruses (HuCV) diarrhea among infants and young children with acute diarrhea in Lanzhou, Gansu Province, China by using molecular epidemiologic techniques. METHODS: Stool specimens were collected from both outpatients and inpatients with acute diarrhea in Lanzhou. Enzyme-linked immunosorbant assay (ELISA) was used to detect rotavirus antigen (RVA). Reverse transcription-polymerase chain reactions (RT-PCR) were used to detect HuCV in stool specimens of RV ELISA (-). RESULTS: Of the stool specimens collected from 515 cases in Lanzhou from December 2001 to June 2004, 264 were RVA ELISA (+) and 251 were RVA ELISA (-). Among all cases who were RVA ELISA (-), 25 (9.96%) were found positive for HuCV. HuCV was detected in 12 of 133 cases (9.02%) from December 2001 to November 2002, no genotyping was performed for these cases. From July 2003 to June 2004 13 of 118 cases (11.02%) were found positive for HuCV, of whom 11 cases had Norwalk-like virus GII (NLV GII) infection and 2 cases had Sapporo-like virus infection (one case had combined infection with astrovirus) and no NLV GI was found. HuCV infection mainly occurred in children under 2 years of age and no seasonal cluster was found. CONCLUSION: HuCV is one of the major etiological agents of viral diarrhea among infants and young children in Lanzhou. NLV GII maybe the predominant genotype.

[Phylogenetic analysis of enterovirus 71 isolated from patients with hand, foot and mouth disease in Guangdong and Fujian provinces, 2000-2001].

OBJECTIVE: To identify enterovirus 71 (EV71) strains isolated from patients with hand, foot and mouth disease (HFMD) in Guangdong and Fujian provinces from 2000 to 2001 by using phylogenetic analysis. METHODS: All 25 samples were first tested for enteric viruses by RT-PCR using enterovirus specific primers EV-1 and EV-2, and then were identified for EV71 by RT-PCR using EV71 specific primers 159S and 162A. The amplicons of 485bp segment (part of the VP1 gene) were cloned into pGEM-T and sequenced. A phylogenetic tree was constructed by comparison of the sequences with other 12 EV71 strains isolated from China, Japan, Hungary, and the United States including the prototype BrCr. RESULTS: The positive rate of EV71 was about 20%. The sequence analysis showed that the new isolate (GZH2000) shared 94%-96% nucleotide identity with three strains isolated in 1998 and 2000, and 91% with a strain isolated in 1987 from Chinese mainland, but shared only 82%-84% homology with EV71 isolates studied abroad. CONCLUSIONS: EV71 is one of the important pathogens of HFMD in south China. The strains isolated from mainland were closely related with most isolates from Taiwan, but different from most EV71 strains reported abroad. The symptoms of EV71 infection in mainland were not as intensive as those described in Taiwan's outbreak.