Trend and heterogeneity in forced vital capacity among Chinese students during 1985-2019: results from Chinese National Survey on Students' Constitution and Health.

BACKGROUND: Forced vital capacity (FVC) reflects respiratory health, but the long-term trend and heterogeneity in FVC of Chinese students were understudied. METHODS: Data were from Chinese National Survey on Students' Constitution and Health 1985-2019. Super Imposition by Translation and Rotation model was used to draw FVC growth curves. Sex-, region-, and nationality-heterogeneity in FVC was evaluated. Spearman correlation and generalized additive model was used to reveal influencing factors for FVC. RESULTS: Compared to 1985, age at peak FVC velocity was 1.09, 3.17, 0.74, and 1.87 years earlier for urban male, urban female, rural male, and rural female in 2019, respectively. Peak FVC velocity first decreased and then increased during 1985-2019, only male rebounded to larger than 1985 level. FVC declined from 1985 to 2005 and then raised. Males consistently had higher FVC than females, with disparities increasing in the 13-15 age group. Urban students also had higher FVC than rural students. In 2019, FVC difference between 30 Chinese provinces and the national average showed four scenarios: consistently above national average; less than national average until age 18, then above; greater than national average until age 18, then this advantage reversed; less than national average in almost all the age. Most Chinese ethnic minority students had lower FVC levels compared to Han students. Spearman correlation and generalized additive model showed that age, sex, and height were the leading influencing factors of FVC, followed by socioeconomic and environmental factors. CONCLUSIONS: Chinese students experienced advanced FVC spurt, and there was sex-, region- and nationality-heterogeneity in FVC. Routine measurement of FVC is necessary in less developed areas of China.

Comparative study of trimming and resection of diseased finger superficial flexor tendons in gouty carpal tunnel syndrome.

BACKGROUND: Hyperuricemia can lead to synovial hyperplasia in the wrist. In severe cases, it can lead to the deposition of gouty stone in the carpal tunnel, resulting in increased pressure in the carpal tunnel and compression of the median nerve to cause carpal tunnel syndrome (CTS), which is called gouty carpal tunnel syndrome (GCTS). As for the surgical treatment of gouty carpal tunnel syndrome, scholars have different opinions on whether it is necessary to remove the superficial flexor tendon. The purpose of this study was to compare the clinical efficacy of trimming and resection of the diseased superficial flexor tendon in the treatment of gouty carpal tunnel syndrome. METHODS: Clinical data were collected from May 2016 to July 2021 from 10 patients (13 affected wrists) diagnosed with gouty carpal tunnel syndrome and classified into two groups according to the surgical modality: the diseased portion of the gout-eroded superficial finger tendon was trimmed in 9 wrists, and the diseased superficial finger flexor tendon was excised in 4 wrists. Values related to flexion and extension functions, 2-PD, DASH, BCTQ, VAS and recurrence in the affected fingers were compared between the two groups as well as before and after surgery in each group. RESULTS: All affected limbs used were cleared of gouty stones, finger numbness improved, no skin necrosis occurred, and all incisions healed at stage I. At follow-up (13.58 ± 5.53 months), there was no significant difference between groups in flexion and extension function, 2-PD, DASH, BCTQ, and VAS with respect to the affected fingers, and patients in both groups improved significantly before and after surgery. Treatment of only one wrist involved trimming to remove lesion-affected portions of tendon, which reappeared 1 year after surgery, and there was one case of poor recovery from greater piriformis muscle atrophy in both procedures. CONCLUSION: Regarding surgical treatment of patients with gouty carpal tunnel syndrome in which the gouty stone has invaded the superficial flexor tendons of the fingers, the diseased superficial flexor tendons can be selectively excised, and the postoperative mobility of the affected fingers may not be impaired.

DNMT1 and DNMT3B regulate tumorigenicity of human prostate cancer cells by controlling RAD9 expression through targeted methylation.

Prostate cancer is the second most common type of cancer and the second leading cause of cancer death in American men. RAD9 stabilizes the genome, but prostate cancer cells and tumors often have high quantities of the protein. Reduction of RAD9 level within prostate cancer cells decreases tumorigenicity of nude mouse xenographs and metastasis phenotypes in culture, indicating that RAD9 overproduction is essential for the disease. In prostate cancer DU145 cells, CpG hypermethylation in a transcription suppressor site of RAD9 intron 2 causes high-level gene expression. Herein, we demonstrate that DNA methyltransferases DNMT1 and DNMT3B are highly abundant in prostate cancer cells DU145, CWR22, LNCaP and PC-3; yet, these DNMTs bind primarily to the transcription suppressor in DU145, the only cells where methylation is critical for RAD9 regulation. For DU145 cells, DNMT1 or DNMT3B shRNA reduced RAD9 level and tumorigenicity, and RAD9 ectopic expression restored this latter activity in the DNMT knockdown cells. High levels of RAD9, DNMT1, DNMT3B and RAD9 transcription suppressor hypermethylation were significantly correlated in prostate tumors, and not in normal prostate tissues. Based on these results, we propose a novel model where RAD9 is regulated epigenetically by DNMT1 and DNMT3B, via targeted hypermethylation, and that consequent RAD9 overproduction promotes prostate tumorigenesis.

Characteristics, Sources, and Risks of Polycyclic Aromatic Hydrocarbons in Topsoil and Surface Water from the Liuxi River Basin, South China.

The concentrations, composition, sources, and risks of polycyclic aromatic hydrocarbons (PAHs) in topsoil and surface water of the Liuxi River basin, south China were analyzed in this study. The total concentrations of 16 PAHs ranged from 296.26 to 888.14 ng/g in topsoil and from 156.73 to 422.03 ng/L in surface water, indicating mild pollution. The PAHs in topsoil exhibited an even spatial distribution, suggesting that they originated primarily from dry and wet deposition of transported pollutants. The concentration of PAHs in surface water did not differ significantly geographically, but the concentrations of total, three-, and four-ring PAHs were significantly lower in the Liuxi River than in its tributaries. Three- and two-ring PAHs predominated in topsoil and surface water, respectively. A correlation analysis suggested that the total organic carbon content and pH exerted a negligible effect on the spatial distribution of PAHs in topsoil, and they may have common sources. Fossil fuel combustion (particularly vehicle emissions) and coking production were the dominant sources of PAHs in topsoil, whereas those in surface water were derived from a variety of sources. The total toxic equivalent concentrations of 16 PAHs in topsoil ranged from 3.73 to 105.66 ng/g (mean, 30.93 ng/g), suggesting that exposure to the basin's topsoil does not pose a risk to the environment or public health according to the Canadian soil quality guidelines. A risk assessment revealed that the total PAH concentrations in surface water posed a low ecological risk.

Seasonal characteristics of chemical compositions and sources identification of PM2.5 in Zhuhai, China.

Fine particulate matter is associated with adverse health effects, but exactly which characteristics of PM2.5 are responsible for this is still widely debated. We evaluated seasonal dynamics of the composition and chemical characteristics of PM2.5 in Zhuhai, China. PM2.5 characteristics at five selected sites within Zhuhai city were analyzed. Sampling began on January 10, 2015, and was conducted for 1 year. The ambient mass concentration, carbon content (organic and elemental carbon, OC and EC), level of inorganic ions, and major chemical composition of PM2.5 were also determined. Average concentrations of PM2.5 were lower than the National Ambient Air Quality Standard (NAAQS) 24-h average of 65 µg/m3. The daily PM2.5 concentration in Zhuhai city exhibited clear seasonal dynamics, with higher daily PM2.5 concentrations in autumn and winter than in spring and summer. Carbon species (OC and EC) and water-soluble ions were the primary components of the PM2.5 fraction of particles. Apart from OC and EC, chemical species in PM2.5 were mainly composed of NH4+ and SO42-. There was a marked difference between the summer and winter periods: the concentrations of OC and EC in winter were roughly 3.4 and 4.0 times than those in summer, while NH4+, SO42-, NO3-, and Na+ were 3.2, 4.5, 28.0, and 5.7 times higher in winter than those in summer, respectively. The results of chemical analysis were consistent with three sources dominating PM2.5: coal combustion, biomass burning, and vehicle exhaust; road dust and construction; and from reaction of HCl and HNO3 with NH3 to form NH4Cl and NH4NO3. However, additional work is needed to improve the mass balance and to obtain the source profiles necessary to use these data for source apportionment.

Uptake of Cd(II) Onto Raw Crab Shells: Isotherm, Kinetic, Adsorption Properties And Mechanisms.

This study focuses on the uptake of cadmium (Cd(II)) ion from water onto raw crab shells acting as a biosorbent. Experimental data were perfectly fitted to the Langmuir and Dubinin-Radushkevich models, while the uptake kinetics are well-described using the pseudo-second-order model. The effects of experimental conditions on the uptake isotherms and kinetics are discussed in detail. A study of uptake thermodynamics has indicated that the Cd(II) uptake onto the crab shell is an endothermic and spontaneous process. For further understanding of this process, possible mechanisms of Cd(II) uptake onto crab shell have been deduced from experimental results. Desorption experiment shows that 93.7% of adsorbed Cd(II) could be recovered with a 0.1 M HNO3 solution. The inhibitory effects of other metal co-ions on the Cd(II) uptake on the crab shell are also discussed.

Fas/FasL, Bcl2 and Caspase-8 gene polymorphisms in Chinese patients with rheumatoid arthritis.

Apoptosis signals are necessary for maintaining homeostasis and an adequate immune response. Dysregulation of apoptosis-related genes in the immune system has an important impact on autoimmune diseases such as rheumatoid arthritis (RA). Thus, we investigated the association between Fas rs2234767 G/A, FasL rs763110 C/T, Bcl2 rs12454712 T/C, Bcl2 rs17757541 C/G, and Caspase-8 rs1035142 G/T polymorphisms and RA susceptibility in a Chinese population. These five single-nucleotide polymorphisms (SNPs) were studied in a Chinese population consisting of 615 patients with RA and 839 controls. Genotyping was performed using a custom-by-design 48-Plex SNP scan TM kit. Furthermore, we undertook a meta-analysis between FasL rs763110 C/T and RA. This study indicated that Fas rs2234767 and Bcl2 rs17757541 polymorphisms were risk factors for RA. No association was observed between FasL rs763110 C/T, Bcl2 rs12454712 T/C, and Caspase-8 rs1035142 G/T polymorphisms and RA in this study. The results of this meta-analysis suggested no significant association between FasL rs763110 C/T and RA. However, stratification analysis of this meta-analysis indicated that FasL rs763110 C/T increased the risk of Caucasian RA patients. In conclusion, this study demonstrated that Fas rs2234767 G/A and Bcl2 rs17757541 T/C polymorphisms might be associated with an increased risk of RA. This meta-analysis revealed that FasL rs763110 C/T was associated with an increased risk of Caucasian RA patients.

Factors Explaining the Use of Web-Based Consultations With Physicians by Young and Middle-Aged Individuals in China: Qualitative Comparative Analysis.

BACKGROUND: It was only upon the occurrence of the COVID-19 pandemic that the demand for web-based consultations with physicians grew at unprecedented rates. To meet the demand, the service environment developed rapidly during the pandemic. OBJECTIVE: This study aimed to identify the current status of the use of web-based consultations with physicians among young and middle-aged Chinese individuals and explore users' perspectives on key factors that influence its use in terms of optimizing benefits and compensating for disadvantages. METHODS: We conducted semistructured interviews with 65 individuals (aged 18 to 60 years) across China between September and October 2022. The interviewees were selected through snowball sampling. They described their experiences of using web-based physician consultations and the reasons for using or not using the service. Based on the Andersen Behavioral Model, a qualitative comparative analysis was used to analyze the factors associated with the use of web-based physician consultations and explore the combinations of these factors. RESULTS: In all, 31 (48%) of the 65 interviewees used web-based consultation services. The singular necessary condition analysis revealed that the complementary role of the service and perceived convenience are necessary conditions for the use of web-based consultation services, and user's confidence in the service was a sufficient condition. Based on the Andersen Behavioral Model, the configuration analysis uncovered 2 interpretation models: an enabling-oriented model and a need-oriented model. The basic combination of the enabling-oriented model included income and perceived convenience. The basic combination of the need-oriented model included complementary role and user's confidence. CONCLUSIONS: Among the factors associated with the use of web-based consultations, perceived convenience, complementary role, and user's confidence were essential factors. Clear instructions on the conduct of the service, cost regulations, provider qualifications guarantee, privacy and safety supervision, the consultations' application in chronic disease management settings, and subsequent visits can promote the positive development of web-based consultations.

A novel extraction-free dual HiFi-LAMP assay for detection of methicillin-sensitive and methicillin-resistant Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is a leading cause of bacteremia and blood stream infections. Methicillin-resistant S. aureus (MRSA) that first appeared in 1961 often caused hospital-acquired infections (HAIs) and community-acquired infections (CAIs) and was associated with high mortality rate. Accurate and rapid point-of-care testing (POCT) of MRSA is crucial for clinical management and treatment of MRSA infections, as well as the prevention and control of HAIs and CAIs. Here, we reported a novel extraction-free dual HiFi-LAMP assay for discriminative detection of methicillin-susceptible S. aureus and MRSA. The dual HiFi-LAMP assay can detect 30 copies/reaction of nuc and mecA genes with detection limits of 147 and 158 copies per 25 µL reaction, respectively. A retrospective clinical evaluation with 107 clinical S. aureus isolates showed both sensitivity and specificity of 100%. A prospective clinical evaluation with 35 clinical samples revealed a specificity of 100% and a sensitivity of 92.3%. The dual HiFi-LAMP assay can detect almost all S. aureus samples (141/142; 99.3%) within 20 min, implying that the entire HiFi-LAMP assay (including sample process) can be completed within 40 min, extremely significantly shorter than 3-5 days by the traditional clinical microbial culture and antibiotic susceptibility testing. The novel extraction-free dual HiFi-LAMP assay can be used as a robust POCT tool to promote precise diagnosis and treatment of MRSA infections in hospitals and to facilitate surveillance of MRSA at hospital and community settings.IMPORTANCEMethicillin-resistant Staphylococcus aureus (MRSA) was associated with high mortality rate and listed as a "priority pathogen" by the World Health Organization. Accurate and rapid point-of-care testing (POCT) of MRSA is critically required for clinical management and treatment of MRSA infections. Some previous LAMP-based POCT assays for MRSA might be questionable due to their low specificity and the lack of appropriate evaluation directly using clinical samples. Furthermore, they are relatively tedious and time-consuming because they require DNA extraction and lack multiplex detection capacity. Here, we reported a novel extraction-free dual HiFi-LAMP assay for discriminative detection of MRSA and methicillin-susceptible S. aureus. The assay has high specificity and sensitivity and can be completed within 40 min. Clinical evaluation with real clinical samples and clinical isolates showed excellent performance with 100% specificity and 92.3%-100% sensitivity. The novel extraction-free assay may be a robust POCT tool to promote precise diagnosis of MRSA infections and facilitate surveillance of MRSA at hospital and community settings.

Rad9 protein contributes to prostate tumor progression by promoting cell migration and anoikis resistance.

Rad9 as part of the Rad9-Hus1-Rad1 complex is known to participate in cell cycle checkpoint activation and DNA repair. However, Rad9 can act as a sequence-specific transcription factor, modulating expression of a number of genes. Importantly, Rad9 is up-regulated in prostate cancer cell lines and clinical specimens. Its expression correlates positively with advanced stage tumors and its down-regulation reduces tumor burden in mice. We show here that transient down-regulation of Rad9 by RNA interference reduces DU145 and PC3 prostate cancer cell proliferation and survival in vitro. In addition, transient or stable down-regulation of Rad9 impairs migration and invasion of the cells. Moreover, stable reduction of Rad9 renders DU145 cell growth anchorage-dependent. It also decreases expression of integrin ß1 protein and sensitizes DU145 and LNCaP cells to anoikis and impairs Akt activation. On the other hand, stable expression of Mrad9, the mouse homolog, in DU145/shRNA Rad9 cells restores migration, invasion, anchorage-independent growth, integrin ß1 expression, and anoikis resistance with a concomitant elevation of Akt activation. We thus demonstrate for the first time that Rad9 contributes to prostate tumorigenesis by increasing not only tumor proliferation and survival but also tumor migration and invasion, anoikis resistance, and anchorage-independent growth.

Effect of soothing liver therapy on oocyte quality and growth differentiation factor-9 in patients undergoing in vitro fertilization and embryo transfer.

OBJECTIVE: To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer (IVF-ET) and to explore its mechanism. METHODS: Fifty-eight women with tubal infertility were randomized into two groups: 30 in an experimental group treated with Xiaoyao powder (Shugan) plus gonadotropin-releasing hormone analog (GnRHa)/follicle-stimulating hormone (FSH)/ human chorionic gonadotropin (hCG) and 28 in the control group who were treated with GnRHa/FSH/ hCG only. The total gonadotropin (Gn) doses required, endometrial thickness, oocyte numbers, high quality embryo production rate and pregnancy rate of the two groups were compared. The concentration of growth differentiation factor-9 (GDF-9) in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse transcription-polymerase chain reaction amplification. RESULTS: In the experimental group, the Gn dose was significantly lower than that in the control group; the endometrial thickness, high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group (all P < 0.05). CONCLUSION: Shugan treatment can improve the pregnancy rate of women with tubal infertility; its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.

Calibration of a serum reference standard for Group B streptococcal polysaccharide conjugate vaccine development using surface plasmon resonance.

Group B streptococcus (GBS) is a leading cause of neonatal morbidity and mortality worldwide. Development of a maternal vaccine to protect newborns through placentally transferred antibody is considered feasible based on the well-established relationship between anti-GBS capsular polysaccharide (CPS) IgG levels at birth and reduced risk of neonatal invasive GBS. An accurately calibrated serum reference standard that can be used to measure anti-CPS concentrations is critical for estimation of protective antibody levels across serotypes and potential vaccine performance. For this, precise weight-based measurement of anti-CPS IgG in sera is required. Here, we report an improved approach for determining serum anti-CPS IgG levels using surface plasmon resonance with monoclonal antibody standards, coupled with a direct Luminex-based immunoassay. This technique was used to quantify serotype-specific anti-CPS IgG levels in a human serum reference pool derived from subjects immunized with an investigational six-valent GBS glycoconjugate vaccine.

Development of poly(vinyl acetate-methylacrylic acid)/chitosan/Fe3O4 nanoparticles for the diagnosis of non-alcoholic steatohepatitis with magnetic resonance imaging.

Non-alcoholic steatohepatitis is a burgeoning health problem. To diagnose NASH with magnetic resonance imaging (MRI), an effective contrast agent, a stable suspension of superparamagnetic Fe(3)O(4) nanoparticles, were newly developed. The negatively charged Fe(3)O(4) nanoparticles were coated with positive chitosan (CS) firstly, and then assembled with poly(vinyl acetate-methylacrylic acid) (P(VAc-MAA)). Transmission electron microscope and dynamic light scattering confirmed that the obtained P(VAc-MAA)/CS/Fe(3)O(4) nanoparticles had a spherical or ellipsoidal morphology with an average diameter in the range of 14-20 nm. The superparamagnetic property and spinel structure of the Fe(3)O(4) nanoparticles were well preserved due to the protection of the P(VAc-MAA)/CS layers on the surface of the Fe(3)O(4) nanoparticles. The in vivo rat experiments confirmed that the P(VAc-MAA)/CS/Fe(3)O(4) nanoparticles were an effective contrast agent for MRI to diagnose NASH.

Titanium doping reduces superoxide dismutase activity, but not oxidase activity, of catalytic CeO(2) nanoparticles.

In this paper we report the enzymatic properties of Ti-doped CeO(2) nanoparticles. The superoxide dismutase activity of Ti-doped nanoparticles is reduced in comparison to undoped nanoceria. In contrast, the oxidase activity of these nanoparticles was unchanged. The change in enzymatic activity was accompanied by a dramatic change in shape to a spherical nanostructure. In addition to reporting a new type of enzymatically-active nanoparticle, Ti-doped cerium oxide nanoparticles are well suited for biological applications.

An enzymatic colorimetric assay for glucose-6-phosphate.

A specific colorimetric assay for the determination of glucose-6-phosphate (G6P) was developed. This assay is based on the oxidation of G6P in the presence of glucose-6-phosphate dehydrogenase (G6PD) and nicotinamide adenine dinucleotide phosphate (NADP(+)); the NADPH thereby generated reduces the tetrazolium salt WST-1 [2-(4-indophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium, monosodium salt] to water-soluble yellow-colored formazan with 1-methoxy-5-methylphenazium methylsulfate (1-mPMS) as an electron carrier. The assay is optimized for reaction buffer pH, enzyme/dye concentration, and reaction time course. The limit of detection of the assay is 0.15 µM (15 pmol/well). The usefulness of the assay is demonstrated by the accurate measurement of the G6P concentration in fetal bovine serum (FBS).

Mouse Rad9b is essential for embryonic development and promotes resistance to DNA damage.

RAD9 participates in promoting resistance to DNA damage, cell cycle checkpoint control, DNA repair, apoptosis, embryogenesis, and regulation of transcription. A paralogue of RAD9 (named RAD9B) has been identified. To define the function of mouse Rad9b (Mrad9b), embryonic stem (ES) cells with a targeted gene deletion were constructed and used to generate Mrad9b mutant mice. Mrad9b(-/-) embryos are resorbed after E7.5 while some of the heterozygotes die between E12.5 and a few days after birth. Mrad9b is expressed in embryonic brain and Mrad9b(+/-) embryos exhibit abnormal neural tube closure. Mrad9b(-/-) mouse embryonic fibroblasts are not viable. Mrad9b(-/-) ES cells are more sensitive to gamma rays and mitomycin C than Mrad9b(+/+) controls, but show normal gamma-ray-induced G2/M checkpoint control. There is no evidence of spontaneous genomic instability in Mrad9b(-/-) cells. Our findings thus indicate that Mrad9b is essential for embryonic development and mediates resistance to certain DNA damaging agents.

Release characteristics and mechanisms of sediment phosphorus in contaminated and uncontaminated rivers: A case study in South China.

Phosphorus (P) cycling present in sediments associated with iron (Fe), manganese (Mn) and sulfur (S) geochemical processes may cause secondary pollution in overlying water. Understanding the mechanisms of P release from sediments should help to restore water quality. This study used the diffusive gradients in thin film (DGT) technique to investigate the seasonal variation in the lability, remobilization mechanisms, and release characteristics of sediment P in the uncontaminated Xizhi River and the severely contaminated Danshui River, South China. P accumulation in sediments contributed to higher DGT-labile P concentrations in contaminated reaches, and the highest labile P concentrations were generally observed in summer season at each site. The significant positive relationships (p < 0.05) between labile Fe and P confirmed the Fe-P coupling release mechanism in uncontaminated sediments. Stronger relationships between labile Mn and P at contaminated sites indicated that Mn oxides played an important role in P remobilization. However, sulfate reduction associated with microbial activities (crucial genera: Desulfobulbus, Desulfomicrobium and Desulforhabdus) was considered to decouple the Fe & Mn-P cycling relationship, promoting P release at contaminated sites. The effluxes of sediment P were much higher in the Danshui River (mean 0.132 mg cm-2·d-1) than in the Xizhi River (mean 0.038 mg cm-2·d-1). And hot season led to growth in P effluxes that was much greater in contaminated river.

A sensitive fluorimetric assay for pyruvate.

A sensitive and specific fluorimetric assay for the determination of pyruvate is reported here. This assay is based on the oxidation of pyruvate in the presence of pyruvate oxidase. Hydrogen peroxide generated by pyruvate oxidase reacts with nonfluorescent Amplex Red at a 1:1 stoichiometry to form the fluorescent product, resorufin. The assay is optimized with respect to pH of reaction buffer, enzyme concentration, dye concentration, and the time course. The usefulness of the assay is demonstrated by the accurate measurement of intracellular and extracellular pyruvate concentrations. The limit of detection of the assay is 5 nM.

Amplex UltraRed enhances the sensitivity of fluorimetric pyruvate detection.

Pyruvate, which is produced in the final step of glycolysis, participates in anabolic metabolism, catabolic metabolism, and signal transduction. We have recently reported a new sensitive and selective fluorimetric assay for pyruvate measurement using Amplex Red as a fluorogenic dye. However, the fluorescence of the reaction product, resorufin, is pH dependent, which limits the sensitivity of the assay at pH 6.7. In this Note, we evaluate Invitrogen's new dye, Amplex UltraRed, as a fluorogenic agent for pyruvate determination. Our results show that Amplex UltraRed improves the performance of the assay, providing brighter fluorescence and enhanced sensitivity.

Intranasal immunization with chitosan/pCAGGS-flaA nanoparticles inhibits Campylobacter jejuni in a White Leghorn model.

Campylobacter jejuni is the most common zoonotic bacterium associated with human diarrhea, and chickens are considered to be one of the most important sources for human infection, with no effective prophylactic treatment available. We describe here a prophylactic strategy using chitosan-DNA intranasal immunization to induce specific immune responses. The chitosan used for intranasal administration is a natural mucus absorption enhancer, which results in transgenic DNA expression in chicken nasopharynx. Chickens immunized with chitosan-DNA nanoparticles, which carried a gene for the major structural protein FlaA, produced significantly increased levels of serum anti-Campylobacter jejuni IgG and intestinal mucosal antibody (IgA), compared to those treated with chitosan-DNA (pCAGGS). Chitosan-pCAGGS-flaA intranasal immunization induced reductions of bacterial expellation by 2-3 log(10) and 2 log(10) in large intestine and cecum of chickens, respectively, when administered with the isolated C. jejuni strain. This study demonstrated that intranasal delivery of chitosan-DNA vaccine successfully induced effective immune response and might be a promising vaccine candidate against C. jejuni infection.