Poor accuracy of single serological IgM tests in children with suspected acute Mycoplasma pneumoniae infection in Guangzhou, China.

Introduction. Early and accurate diagnosis of Mycoplasma pneumoniae (MP) infection of children with pneumonia is at the core of treatment in clinical practice.Gap Statement. Serological immunoglobulin M (IgM) tests for MP infection of children in south China have been rarely described.Aim. To assess the diagnostic performance and clinical application of serodiagnosis of MP infection in paediatric pneumonia patients.Methodology. Serum samples from 144 children diagnosed with MP pneumonia were subjected to a particle agglutination (PA)-based IgM assay. Meanwhile, we used an established suspension array as the reference standard method for the detection of MP DNA in bronchoalveolar lavage fluid (BALF) from all patients to assess the reliability of serological assays.Results. When running immunological testing in single serum samples, 80.6 %(79/98) of cases were diagnosed with MP infection, whereas only 55 (56.1 %) cases were positive in MP DNA analysis. Furthermore, single serum tests for IgM during acute MP infection resulted in 85.5 % (47/55) sensitivity and 25.6 % (11/43) specificity. Nevertheless, immunological testing and MP DNA analysis yielded the same results when paired sera were available for MP IgM antibody testing.Conclusion. Paired serological IgM assays are necessary for the determination of an acute MP infection, whereas single serological IgM testing is unreliable. Moreover, even a short interval of two MP serological tests works well.

CXCR4-targeted nitric oxide nanoparticles deliver PD-L1 siRNA for immunotherapy against glioblastoma.

While immunotherapy has emerged as a promising strategy to treat glioblastoma multiforme (GBM), the limited availability of immunotherapeutic agents in tumors due to the presence of the blood-brain barrier (BBB) and immunosuppressive tumor microenvironment dampens efficacy. Nitric oxide (NO) plays a role in modulating both the BBB and tumor vessels and could thus be delivered to disrupt the BBB and improve the delivery of immunotherapeutics into GBM tumors. Herein, we report an immunotherapeutic approach that utilizes CXCR4-targeted lipid­calcium-phosphate nanoparticles with NO donors (LCP-NO NPs). The delivery of NO resulted in enhanced BBB permeability and thus improved gene delivery across the BBB. CXCR4-targeted LCP-NO NPs delivered siRNA against the immune checkpoint ligand PD-L1 to GBM tumors, silenced PD-L1 expression, increased cytotoxic T cell infiltration and activation in GBM tumors, and suppressed GBM progression. Thus, the codelivery of NO and PD-L1 siRNA by these CXCR4-targeted NPs may serve as a potential immunotherapy for GBM.

A Lateral Flow Immunoassay Coupled with a Spectrum-Based Reader for SARS-CoV-2 Neutralizing Antibody Detection.

As of August 2021, there have been over 200 million confirmed case of coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus and more than 4 million COVID-19-related deaths globally. Although real-time polymerase chain reaction is considered to be the primary method of detection for SARS-CoV-2 infection, the use of serological assays for detecting COVID-19 antibodies has been shown to be effective in aiding with diagnosis, particularly in patients who have recovered from the disease and those in later stages of infection. Since it has a high detection rate and few limitations compared to conventional enzyme-linked immunosorbent assay protocols, we used a lateral flow immunoassay as our diagnostic tool of choice. Since lateral flow immunoassay results interpreted by the naked eye may lead to erroneous diagnoses, we developed an innovative, portable device with the capacity to capture a high-resolution reflectance spectrum as a means of promoting diagnostic accuracy. We combined this spectrum-based device with commercial lateral flow immunoassays to detect the neutralizing antibody in serum samples collected from 30 COVID-19-infected patients (26 mild cases and four severe cases). The results of our approach, lateral flow immunoassays coupled with a spectrum-based reader, demonstrated a 0.989 area under the ROC curve, 100% sensitivity, 95.7% positive predictive value, 87.5% specificity, and 100% negative predictive value. As a result, our approach exhibited great value for neutralizing antibody detection. In addition to the above tests, we also tested plasma samples from 16 AstraZeneca-vaccinated (ChAdOx1nCoV-19) patients and compared our approach and enzyme-linked immunosorbent assay results to see whether our approach could be applied to vaccinated patients. The results showed a high correlation between these two approaches, indicating that the lateral flow immunoassay coupled with a spectrum-based reader is a feasible approach for diagnosing the presence of a neutralizing antibody in both COVID-19-infected and vaccinated patients.

[Interactions of Fungi Community and Relationship with the Carbon Structure in Arable Mollisols with Gradient Organic Matter Content].

Fungi play an important role in the accumulation and transformation of soil organic matter (SOM) and nutrient cycling. To investigate the relationship between the fungal community and soil organic carbon functional groups under gradient SOM contents in arable mollisols, arable mollisols with 2%-9% SOM content were collected in Northeast China. Solid-state 13C-NMR technology was used to explore the differences in the functional group structure of SOM, and ITS high-throughput sequencing was used to investigate the fungal community structure. The potential interactions between different taxonomic groups of soil fungal community and their associations with organic carbon molecular structures were compared by constructing molecular ecological networks under low SOM (2%-5%) and high SOM (7%-9%) conditions. The 13C-NMR results indicated an increase in the relative abundance of Alkyl C (25.8% to 35.9%). The decrease in Alkyl C/O-Alkyl C indicated a smaller degree of decomposition in high SOM soils. Sordariomycetes and Mortierellomycotina dominated the fungal community and their relative abundance increased with the SOM gradient (P<0.05) from 14.33% to 28.17% and from 7.32% to 23.14%, respectively. The network analysis showed simpler ecological topological properties of the fungal community in low SOM soils, with lower numbers of nodes, edges, and average clustering coefficients than those in high SOM soils. A closer relationship between fungi and organic carbon functional groups, especially LOC, was observed in low SOM soils. The random forest model showed that LOC had the largest amount for fungal interactions in low SOM soils (10%), followed by recalcitrant organic carbon (ROC). In comparison, LOC contributed less to the variations in fungal interactions in high SOM soils (7.4%). With globally increasing soil carbon loss, the limition of the carbon resources, especially the reduction of LOC, may reduce the stability and ecological functions of soil fungal communities.

Highly efficient and tumor-selective nanoparticles for dual-targeted immunogene therapy against cancer.

While immunotherapy holds great promise for combating cancer, the limited efficacy due to an immunosuppressive tumor microenvironment and systemic toxicity hinder the broader application of cancer immunotherapy. Here, we report a combinatorial immunotherapy approach that uses a highly efficient and tumor-selective gene carrier to improve anticancer efficacy and circumvent the systemic toxicity. In this study, we engineered tumor-targeted lipid-dendrimer-calcium-phosphate (TT-LDCP) nanoparticles (NPs) with thymine-functionalized dendrimers that exhibit not only enhanced gene delivery capacity but also immune adjuvant properties by activating the stimulator of interferon genes (STING)-cGAS pathway. TT-LDCP NPs delivered siRNA against immune checkpoint ligand PD-L1 and immunostimulatory IL-2-encoding plasmid DNA to hepatocellular carcinoma (HCC), increased tumoral infiltration and activation of CD8+ T cells, augmented the efficacy of cancer vaccine immunotherapy, and suppressed HCC progression. Our work presents nanotechnology-enabled dual delivery of siRNA and plasmid DNA that selectively targets and reprograms the immunosuppressive tumor microenvironment to improve cancer immunotherapy.

Encapsulation and sustained release of curcumin by a composite hydrogel of lotus root amylopectin and chitosan.

In the present work, lotus root amylopectin (LRA)-chitosan (CS) composite hydrogel was developed as a delivery system for curcumin (CUR). Results exhibited that a stable LRA-CS-CUR hydrogel was formed using LRA and CS in the ratio of 3:2 (w/w) at pH 4.0. Under this condition, the particle size, polydispersity index and zeta potential of the LRA-CS-CUR was 410.3 nm, 0.211 and +26.47 mV, respectively. The analysis of transmission electron microscopy, fourier transform infrared spectroscopy and X-ray diffractometer revealed that curcumin was successfully encapsulated in the LRA-CS hydrogel, giving a high encapsulation efficiency of 90.3 %. Moreover, this composite hydrogel could significantly improve the solubility and stability of curcumin. The release characteristics of encapsulated curcumin in simulated gastric fluid and simulated intestinal fluid were further investigated. Results exhibited that the LRA-CS hydrogel enabled curcumin to be stable in stomach and release in small intestine.

Galactose Derivative-Modified Nanoparticles for Efficient siRNA Delivery to Hepatocellular Carcinoma.

Successful siRNA therapy requires suitable delivery systems with targeting moieties such as small molecules, peptides, antibodies, or aptamers. Galactose (Gal) residues recognized by the asialoglycoprotein receptor (ASGPR) can serve as potent targeting moieties for hepatocellular carcinoma (HCC) cells. However, efficient targeting to HCC via galactose moieties rather than normal liver tissues in HCC patients remains a challenge. To achieve more efficient siRNA delivery in HCC, we synthesized various galactoside derivatives and investigated the siRNA delivery capability of nanoparticles modified with those galactoside derivatives. In this study, we assembled lipid/calcium/phosphate nanoparticles (LCP NPs) conjugated with eight types of galactoside derivatives and demonstrated that phenyl ß-d-galactoside-decorated LCP NPs (L4-LCP NPs) exhibited a superior siRNA delivery into HCC cells compared to normal hepatocytes. VEGF siRNAs delivered by L4-LCP NPs downregulated VEGF expression in HCC in vitro and in vivo and led to a potent antiangiogenic effect in the tumor microenvironment of a murine orthotopic HCC model. The efficient delivery of VEGF siRNA by L4-LCP NPs that resulted in significant tumor regression indicates that phenyl galactoside could be a promising HCC-targeting ligand for therapeutic siRNA delivery to treat liver cancer.

Effects of folic acid on the performance of suckling piglets and sows during lactation.

BACKGROUND: The present research was conducted to investigate the influences of supplementation with different levels of folic acids in diet on the performance of lactating sows. Twenty Landrace × Yorkshire sows received the same basal corn-soybean diets (folic acid, 1.3 mg kg(-1)) from gestation to parturition (day 107). After parturition, sows were allotted to four treatments: control group (folic acid supplementation level, 0 mg kg(-1)), group 1 (12.5 mg kg(-1)), group 2 (50 mg kg(-1)) and group 3 (100 mg kg(-1)), with five replicates of one sow. The experiment lasted for 21 days. RESULTS: (1) Folic acid increased milk production (P > 0.05). (2) Compared with the control, supplementation with folic acid (100 mg kg(-1)) increased the concentration of butter fat, total substance and non-lipoid substance significantly (P < 0.01), the concentration of milk protein was also significantly increased in group 2 and group 3 (P < 0.01) in milk. (3) Folic acid supplementation could increase litter weaning weight, average piglet weaning weight and average piglet daily gain (P > 0.05). CONCLUSION: These results suggested that supplementation with folic acid in the diets of lactating sows increases milk production, improved milk quality and the performance of piglets.

Effect of polysaccharides of cassiae seeds on the intestinal microflora of piglets.

The objective of this study was to examine if polysaccharides from Cassiae Seeds (PCS) can be used as prebiotics to improve the intestinal microflora of piglets with an in vitro and an in vivo trial. The in vitro trial was conducted to study the dose-response effect of PCS on the growth of E. coli 09 and Lactobacillus with traditional plate count method. The gradient culture mediums, containing 3.2, 1.6, 0.8, 0.4, 0.2, 0.1, 0.05, 0.025 and 0% PCS, were inoculated with E. coli09, Lactobacillus and cecum content, respectively. PCS had no influence on the growth of E. coli09 from rejuvenation fluid, but inhibited the growth of E. coli09 from cecum content when the concentration of PCS was higher than 0.1%. Lactobacillus counts were significantly increased with 0.1% PCS or higher (p< 0.05); and the largest increase was found with 0.8% PCS. With the inoculum of cecum content in the medium, Lactobacillus counts increased when the concentration of PCS was 0.4% and 0.8%, whilst E. coli 09 counts decreased. The in vivo trial was carried out to investigate the effect of PCS on the growth of E. coli 09 and Lactobacillus in piglets. Thirty six barrows (average initial BW = 6.5 kg) were randomly divided into 3 groups with 6 each, fed diets supplemented without or with 0.4% or 0.8% PCS. After 14 days, 3 piglets were slaughtered from each group; digesta samples were collected from the ileum, cecum and colon for detection of E.coli 09 and Lactobacillus with plate count method. Samples of the tissue and content of the cecum were taken for detection of caecal microflora profiles with Denaturing Gradient Gel Electrophoresis (DGGE) technique. The dietary inclusion of PCS increased Lactobacillus counts, but reduced E. coli 09 counts in digesta of ileum, cecum and colon of piglets. The dietary inclusion of 0.8% PCS significantly increased the number of electrophoresis brands of caecal bacterial microflora in mucosa and content of the cecum (p< 0.05). These results confirmed the dynamic change in the intestinal microflora profile with the dietary inclusion of PCS in piglets. Thus, PCS can be used as prebiotics to improve the intestinal microflora.

Effect of dietary supplementation of chitosan and galacto-mannan-oligosaccharide on serum parameters and the insulin-like growth factor-I mRNA expression in early-weaned piglets.

The study was to determine effects of dietary supplementation of chitosan (COS) and galacto-mannan-oligosaccharides (GMOS) on some serum biochemical indices, serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) levels, and hepatic and long gissimus muscle IGF-I mRNA expression in early-weaned piglets. Twenty six Duroc x Landrace x Yorkshire piglets at the age of 15 days were used. The piglets had access to creep feed during the suckling. Six piglets were sacrificed for sampling at the beginning of the study. The other 20 piglets were individually housed in metabolic cages and randomly allotted to four corn and soybean meal-based diets including the control group, the antibiotic group with 110 mg lincomycin/kg diet, the COS group containing 0.025% COS, and the GMOS group with 0.20% GMOS, respectively, in a 2-week feeding experiment. Blood urea nitrogen (BUN) level was reduced whereas serum total protein concentration was increased (P<0.05) in responses to the COS and GMOS supplementation. Dietary supplementation of COS and GMOS also increased (P<0.05) the serum GH and IGF-I levels along with enhanced hepatic and the muscle IGF-I mRNA abundance. Dietary supplementation of oligosaccharides such as COS and GMOS may improve growth and feed conversion efficiency by increasing plasma GH and IGF-I levels, in the early-weaned piglets.