High altitude air pollution and respiratory disease: Evaluating compounded exposure events and interactions.

Today, air pollution remains a significant issue, particularly in high-altitude areas where its impact on respiratory disease remains incompletely explored. This study aims to investigate the association between various air pollutants and outpatient visits for respiratory disease in such regions, specifically focussing on Xining from 2016 to 2021. By analysing over 570,000 outpatient visits using a time-stratified case-crossover design and conditional logistic regression, we assessed the independent effects of pollutants like PM2.5, PM10, SO2, NO2, and CO, as well as their interactions. The evaluation of interactions employed measures such as relative excess odds due to interaction (REOI), attributable proportion due to interaction (AP), and synergy index (S). We also conducted a stratified analysis to identify potentially vulnerable populations. Our findings indicated that exposure to PM2.5, PM10, SO2, NO2, and CO significantly increased outpatient visits for respiratory disease, with odds ratios (ORs) of 2.40 % (95 % CI: 2.05 %, 2.74 %), 1.07 % (0.98 %, 1.16 %), 3.86 % (3.23 %, 4.49 %), 4.45 % (4.14 %, 4.77 %), and 6.37 % (5.70 %, 7.04 %), respectively. However, exposure to O3 did not show a significant association. We found significant interactions among PM2.5, SO2, NO2, and CO, where combined exposure further exacerbated the risk of respiratory diseases. For example, in the combination of PM2.5 and SO2, the REOI, AP, and S were 0.07 (95 % CI: 0.06, 0.09), 0.07 (0.06, 0.07), and 1.07 (1.05, 1.09), respectively. Additionally, elderly individuals and females were more sensitive to these pollutants, but no statistically significant interaction effects were observed between different age and gender groups. In conclusion, our study highlights the strong link between air pollution and respiratory disease in high-altitude areas, with combined pollutant exposure posing an even greater risk. It underscores the need for enhanced air quality monitoring and public awareness campaigns, particularly to protect vulnerable populations like the elderly and females.

Additives improve the fermentation quality, anthocyanin content, and biological activity of purple Napier grass silage.

BACKGROUND: Purple Napier grass (PNG), a widely used grass rich in anthocyanin, is commonly employed in the production of silage. However, there is currently limited research on the retention of anthocyanin with or without additives during ensiling. Therefore, this study aimed to investigate the effect of different additives (Lactiplantibacillus plantarum CCZZ1 (LP), glucose, acetic acid, and dried soybean curd residue) on fermentation quality, anthocyanin content, and microbial community structure of PNG silage. RESULTS: Ensiling PNG without additives led to poor fermentation quality and rapid degradation of anthocyanin, resulting in a decline in antioxidant activity and the persistence of harmful microorganisms with high relative abundance. The use of additives, especially LP, effectively increased the relative abundance of L. plantarum, enhancing fermentation quality, the retention of anthocyanin (up to 166% increase rate) and antioxidant activity, while reducing the relative abundance of harmful microorganisms during ensiling for 30 days. Additionally, prolonged ensiling negatively affected the preservation of anthocyanin. Based on both fermentation quality and bioactivity, PNG should be ensiled for 30 days with LP inoculation. CONCLUSION: The employment of additives, especially LP, improved the fermentation quality, anthocyanin retention, and microbial community structure in PNG silage. To optimize both fermentation quality and bioactivity, it is recommended that PNG be ensiled for 30 days with LP inoculation. © 2024 Society of Chemical Industry.

Effects of combined exposure to fine particulate matter and cold waves and on IHD hospitalizations at low and high altitudes.

Climate change and air pollution are major challenges facing the world today. Cold waves and air pollution significantly impact ischemic heart disease (IHD), but the extent of these effects at different altitudes remains unclear, especially their interactions. We collected daily meteorological, pollutant, and IHD hospitalization data from Xining and Xinxiang from 2016 to 2021. Using a time-stratified case-crossover approach, we fitted conditional Poisson regression models to assess the association between cold waves, PM2.5, and IHD hospitalizations and quantified their interactions. Additionally, we calculated the attributable fraction (AF) and attributable number (AN) of hospitalizations due to exposure to cold waves and medium to high-level PM2.5. We also performed stratified analyses by altitude, gender, and age. Both cold waves and PM2.5 were positively associated with IHD hospitalization rates in Xining and Xinxiang, but the differences between the two regions were not significant. The relative risk of cold waves was 1.15 (1.07, 1.24) in Xining and 1.16 (1.11, 1.21) in Xinxiang. In Xining, there was an interaction between cold waves and different levels of PM2.5. We estimated the attributable fraction due to the joint exposure of cold waves and PM2.5 to be 0.14-0.49 in Xining and 0.26-0.36 in Xinxiang. Older adults and males faced higher risks. This study highlights the importance of reducing PM2.5 exposure and optimizing extreme weather warning systems and suggests further exploration of the impacts of individual behaviors and regional characteristics on IHD.

A study on the mediating role of serum hormones in the effects of heavy metals on preeclampsia.

Exposure to environmental heavy metals may pose a risk factor for developing preeclampsia (PE) modified through intervention. This case-control study aimed to investigate the association between serum heavy metal concentrations and PE in pregnant women and whether hormones served as mediating factors in the impact of heavy metals on PE. From October 2020 to 2022, 160 patients with PE and 160 pregnant women with normal deliveries were recruited at Dongguan Songshan Lake Central Hospital. Serum concentrations of manganese (Mn), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), lead (Pb), ß-human chorionic gonadotropin (ß-hCG), progesterone (P), estradiol (E2), testosterone (T), cortisol (Cort), and cortisone (Cor) were measured. Logistic, restricted cubic splines, weighted quantile sum and multivariate linear regression models were employed to account for different aspects and explore the relationships among heavy metals, hormones, and PE. Mediation model analysis was performed to assess the role of hormones in mediation. The median concentrations of Mn, E2, and Cort were lower in the PE group than in the control group. The median concentrations of Cu, Zn, ß-hCG, and T were higher in the PE than in the control. Mn, E2, and Cort showed negative associations with PE, while Cu, Zn, ß-hCG, and T demonstrated positive associations, as determined through logistic regression. Mn, Cu, and Zn displayed linear dose-response relationships with PE. Zn and Cu had high weights in the positive association model of mixed heavy metal exposure with PE. The mediation analysis revealed that serum E2, P, T, Cort, and Cort/Cor might be potential mediators of the association between heavy metals (Mn, Cu, and Zn) and PE.

The mediating role of steroid hormones in the relationship between bisphenol A and its alternatives bisphenol S and F exposure and preeclampsia.

Steroid hormone imbalance is believed to increase the odds of developing PE. Bisphenol A (BPA) and its substitutes (e.g., bisphenol S (BPS) and bisphenol F (BPF)) have estrogen-like effects, and its exposure may be related to the development of preeclampsia (PE). To explore the effects of bisphenol exposure on maternal serum steroid hormones and the potential mediating role of steroid hormones in the association between bisphenol exposure and developing PE, concentrations of bisphenols and steroid hormones in serum samples of 383 pregnant women were examined before delivery (including 160 PE cases and 223 control cases). Multivariable logistic and linear models were used to explore the associations of maternal serum bisphenols concentrations with both maternal steroid hormones and PE risk. Mediation modeling was employed to evaluate the mediating role of steroid hormones in the association between bisphenols and PE. Results showed that maternal serum BPS concentrations were positively associated with testosterone (T) concentrations. The mediation analyses suggested that approximately 10.17 % of the associations between BPS concentrations and the development of PE might be mediated by maternal T. In conclusion, maternal exposure to BPS during pregnancy is linked to higher maternal T concentrations, which might increase the odds of developing PE. T might mediate the association between BPS exposure and the development of PE.

Effects of the interaction between cold spells and fine particulate matter on mortality risk in Xining: a case-crossover study at high altitude.

Background: With global climate change, the health impacts of cold spells and air pollution caused by PM2.5 are increasingly aggravated, especially in high-altitude areas, which are particularly sensitive. Exploring their interactions is crucial for public health. Methods: We collected time-series data on meteorology, air pollution, and various causes of death in Xining. This study employed a time-stratified case-crossover design and conditional logistic regression models to explore the association between cold spells, PM2.5 exposure, and various causes of death, and to assess their interaction. We quantitatively analyzed the interaction using the relative excess odds due to interaction (REOI), attributable proportion due to interaction (AP), and synergy index (S). Moreover, we conducted stratified analyses by average altitude, sex, age, and educational level to identify potential vulnerable groups. Results: We found significant associations between cold spells, PM2.5, and various causes of death, with noticeable effects on respiratory disease mortality and COPD mortality. We identified significant synergistic effects (REOI>0, AP > 0, S > 1) between cold spells and PM2.5 on various causes of death, which generally weakened with a stricter definition of cold spells and longer duration. It was estimated that up to 9.56% of non-accidental deaths could be attributed to concurrent exposure to cold spells and high-level PM2.5. High-altitude areas, males, the older adults, and individuals with lower educational levels were more sensitive. The interaction mainly varied among age groups, indicating significant impacts and a synergistic action that increased mortality risk. Conclusion: Our study found that in high-altitude areas, exposure to cold spells and PM2.5 significantly increased the mortality risk from specific diseases among the older adults, males, and those with lower educational levels, and there was an interaction between cold spells and PM2.5. The results underscore the importance of reducing these exposures to protect public health.

Cold waves and fine particulate matter in high-altitude Chinese cities: assessing their interactive impact on outpatient visits for respiratory disease.

BACKGROUND: Extreme weather events like heatwaves and fine particulate matter (PM2.5) have a synergistic effect on mortality, but research on the synergistic effect of cold waves and PM2.5 on outpatient visits for respiratory disease, especially at high altitudes in climate change-sensitive areas, is lacking. METHODS: we collected time-series data on meteorological, air pollution, and outpatient visits for respiratory disease in Xining. We examined the associations between cold waves, PM2.5, and outpatient visits for respiratory disease using a time-stratified case-crossover approach and distributional lag nonlinear modeling. Our analysis also calculated the relative excess odds due to interaction (REOI), proportion attributable to interaction (AP), and synergy index (S). We additionally analyzed cold waves over time to verify climate change. RESULTS: Under different definitions of cold waves, the odds ratio for the correlation between cold waves and outpatient visits for respiratory disease ranged from 0.95 (95% CI: 0.86, 1.05) to 1.58 (1.47, 1.70). Exposure to PM2.5 was significantly associated with an increase in outpatient visits for respiratory disease. We found that cold waves can synergize with PM2.5 to increase outpatient visits for respiratory disease (REOI > 0, AP > 0, S > 1), decreasing with stricter definitions of cold waves and longer durations. Cold waves' independent effect decreased over time, but their interaction effect persisted. From 8.1 to 21.8% of outpatient visits were due to cold waves and high-level PM2.5. People aged 0-14 and ≥ 65 were more susceptible to cold waves and PM2.5, with a significant interaction for those aged 15-64 and ≥ 65. CONCLUSION: Our study fills the gap on how extreme weather and PM2.5 synergistically affect respiratory disease outpatient visits in high-altitude regions. The synergy of cold waves and PM2.5 increases outpatient visits for respiratory disease, especially in the elderly. Cold wave warnings and PM2.5 reduction have major public health benefits.

Health impacts of a cold wave and its economic loss assessment in China's high-altitude city, Xining.

OBJECTIVE: Amidst climate change, extensive research has centered on the health impacts of heatwaves, yet the consequences of cold spells, particularly in cooler, higher-altitude regions, remain under-explored. METHODS: Analyzing climatic data and non-accidental mortality in Xining, China's second-highest provincial capital, from 2016 to 2020, this study defines cold spells as daily mean temperatures below the 10th, 7.5th, or 5th percentiles for 2-4 consecutive days. A time-stratified case-crossover approach and distributional lag nonlinear modeling were used to assess the link between cold spells and mortality, calculating attributable fractions (AFs) and numbers (ANs) of deaths. The study also examined the impact of cold spells over different periods and analyzed the value of a statistical life (VSL) loss in 2018, a year with frequent cold spells. Stratified analyses by sex, age, and education level were conducted. RESULTS: A significant association was found between cold spells and non-accidental mortality, with a relative risk of 1.548 (95% CI: 1.300, 1.845). The AF was 33.48%, with an AN of 9,196 deaths during the study's cold period. A declining trend in mortality risk was observed from 2019-2020. The 2018 VSL was approximately 2.875 billion CNY, about 1.75% of Xining's GDP. Higher risks were noted among males, individuals aged ≥ 65, and those with lower education levels. CONCLUSION: The findings underscore the vulnerability and economic losses of high-altitude cities to cold spells. Implementing interventions such as improved heating, educational programs, and community support is vital for mitigating these adverse health effects.

An exploratory study on the association of multiple metals in serum with preeclampsia.

Background: Individual metal levels are potential risk factors for the development of preeclampsia (PE). However, understanding of relationship between multiple metals and PE remains elusive. Purpose: The purpose of this study was to explore whether eight metals [zinc (Zn), manganese (Mn), copper (Cu), nickel (Ni), lead (Pb), arsenic (As), cadmium (Cd), and mercury (Hg)] in serum had a certain relationship with PE. Methods: A study was conducted in Dongguan, China. The concentrations of metals in maternal serum were assessed using inductively coupled plasma mass spectrometry (ICP-MS). Data on various factors were collected through a face-to-face interview and hospital electronic medical records. The unconditional logistic regression model, principal component analysis (PCA) and Bayesian Kernel Machine Regression (BKMR) were applied in our study. Results: The logistic regression model revealed that the elevated levels of Cu, Pb, and Hg were associated with an increased risk of PE. According to PCA, principal component 1 (PC1) was predominated by Hg, Pb, Mn, Ni, Cu, and As, and PC1 was associated with an increased risk of PE, while PC2 was predominated by Cd and Zn. The results of BKMR indicated a significant positive cumulative effect of serum metals on PE risk, with Ni and Cu exhibiting a significant positive effect. Moreover, BKMR results also revealed the nonlinear effects of Ni and Cd. Conclusion: The investigation suggests a potential positive cumulative impact of serum metals on the occurrence of PE, with a particular emphasis on Cu as a potential risk factor for the onset and exacerbation of PE. These findings offer valuable insights for guiding future studies on this concern.

Single-use flexible bronchoscopes vs traditional reusable flexible bronchoscopes: a prospective controlled study.

BACKGROUND: Single-use flexible bronchoscopes(SFB) eliminate the risk of bronchoscopy-related infection compared with traditional reusable flexible bronchoscopes(RFB). At present, there is no comparative study between SFB and RFB in the aspects of biopsy and interventional therapy. This study aims to explore whether SFB can perform complex bronchoscopic procedures such as transbronchial biopsies just like RFB. METHODS: We conducted a prospective controlled study. A total of 45 patients who required bronchoscopic biopsy in our hospital from June 2022 to December 2022 were enrolled. The patients were divided into the SFB group and the RFB group, and routine bronchoscopy, bronchoalveolar lavage, and biopsy were performed respectively. Data on the time of routine bronchoscopy, the recovery rate of bronchoalveolar lavage fluid(BALF), biopsy time, and bleeding volume were collected. Then we used the two-sample t-test and the χ2 test to assess the performance differences between SFB and RFB. We also designed a questionnaire to compare the performance between SFB and RFB by different bronchoscope operators. RESULTS: The routine examination time of SFB and RFB was 3.40 ± 0.50 min and 3.55 ± 0.42 min, respectively. There was no significant difference between the two groups (P = 0.308). The recovery rate of BALF was (46.56 ± 8.22) % in the SFB group and (47.00 ± 8.07) in the RFB group, without a significant difference between the two groups(P = 0.863). The biopsy time was similar(4.67 ± 0.51 min VS 4.57 ± 0.45 min) in both groups, with no significant difference(P = 0.512). The positive biopsy rate was 100% in both groups, with no significant difference. Overall, the bronchoscope operators were generally satisfied with SFB. CONCLUSION: SFBs are non-inferior to RFBs in routine bronchoscopy, bronchoalveolar lavage, and biopsy. It is suggested that SFBs have a wider clinical application.

Effects of Hevea brasiliensis Intercropping on the Volatiles of Pandanus amaryllifolius Leaves.

Pandanus amaryllifolius Roxb. is a special tropical spice crop resource with broad development prospects. It is widely cultivated under a Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. canopy to improve the comprehensive benefits to Hevea brasiliensis plantations in Hainan Provence, China. However, the effects of intercropping with Hevea brasiliensis on the component number and relative contents of volatile substances in different categories in the Pandanus amaryllifolius leaves are still unknown. Therefore, a Hevea brasiliensis and Pandanus amaryllifolius intercropping experiment was set up to clarify the differences between several cultivated patterns on volatile substances in the Pandanus amaryllifolius leaves, and the key regulatory factors of volatile substances. The results showed that the soil pH was significantly decreased, while soil bulk density, alkali-hydrolyzable nitrogen and available phosphorus contents were significantly increased under the intercropping pattern. The component numbers of esters in volatile substances were increased by 6.20%, while the component numbers of ketones were decreased by 4.26% under the intercropping pattern. Compared with the Pandanus amaryllifolius monoculture, the relative contents of pyrroles, esters and furanones were significantly increased by 8.83%, 2.30% and 8.27%, respectively, while the relative contents of ketones, furans and hydrocarbons were decreased by 1.01%, 10.55% and 9.16% under the intercropping pattern, respectively. The relative contents of pyrroles, esters, furanones, ketones, furans and hydrocarbons were associated with changes in soil pH, soil available phosphorus content and air temperature. The results indicated that the reduction in soil pH and enhancement in soil-available phosphorus may be the main reasons for promoting the relative content of pyrroles and reducing the relative content of hydrocarbons under an intercropping pattern. Overall, Hevea brasiliensis intercropping with Pandanus amaryllifolius could not only improve soil properties, but also significantly increase the relative contents of the main volatile substances in Pandanus amaryllifolius leaves, which could provide a theoretical basis for the application and promotion of high-quality production patterns of Pandanus amaryllifolius.

Development of alkaline phosphatase-scFv and its use for one-step enzyme-linked immunosorbent assay for His-tagged protein detection.

A histidine (His)-tag is composed of six His residues and typically exerts little influence on the structure and solubility of expressed recombinant fusion proteins. Purification methods for recombinant proteins containing His-tags are relatively well-established, thus His-tags are widely used in protein recombination technology. We established a one-step enzyme-linked immunosorbent assay (ELISA) for His-tagged recombinant proteins. We analyzed variable heavy and light chains of the anti-His-tag monoclonal antibody 4C9 and used BLAST analyses to determine variable zones in light (VL) and heavy chains (VH). VH, VL, and alkaline phosphatase (ALP) regions were connected via a linker sequence and ligated into the pGEX-4T-1 expression vector. Different recombinant proteins with His tags were used to evaluate and detect ALP-scFv activity. Antigen and anti-His-scFv-ALP concentrations for direct ELISA were optimized using the checkerboard method. ZIKV-NS1, CHIKV-E2, SCRV-N, and other His-tag fusion proteins demonstrated specific reactions with anti-His-scFv-ALP, which were accurate and reproducible when the antigen concentration was 50 µg mL-1 and the antibody concentration was 6.25 µg mL-1. For competitive ELISA, we observed a good linear relationship when coating concentrations of recombinant human anti-Müllerian hormone (hAMH) were between 0.78 and 12.5 µg mL-1. Our direct ELISA method is simple, rapid, and accurate. The scFv antibody can be purified using a prokaryotic expression system, which provides uniform product quality and reduces variations between batches.

Associations of maternal blood mercury with preeclampsia and birth outcomes.

BACKGROUND: Mercury (Hg) is a highly toxic substance, and its harmful effects on maternal and infant health have been reported. Yet, the associations of Hg exposure with preeclampsia (PE) and adverse birth outcomes are not well understood. OBJECTIVE: The aim of this study was to investigate the potential effects of maternal Hg exposure on PE and birth outcomes. METHODS: We conducted a case-control study with 84 participants in China. Logistic models were used to estimate odds ratios for PE risk and birth outcomes according to maternal blood Hg levels, adjusting for potential confounding factors. RESULTS: Elevated blood Hg levels were associated with increased risks of mild PE (aOR, 7.03; 95% CI, 1.61, 30.62; P < 0.01) and severe PE (aOR, 47.55; 95% CI, 5.27, 429.05; P < 0.05). We also found that increased blood Hg levels were associated with low birth weight (aOR, 1.12; 95% CI, 1.00, 1.25; P < 0.05) and preterm birth (PTB) (aOR, 1.22; 95% CI, 1.08, 1.38; P < 0.05). CONCLUSIONS: Our study provided evidence that elevated blood Hg levels were significantly associated with an increased risk of PE. In addition, our findings support that increased blood Hg levels might be associated with low birth weight and PTB.

The Relationship Between Preeclampsia and Arsenic Concentration in the Peripheral Blood.

Preeclampsia (PE) is a pregnancy-specific disorder, which is one of the leading causes of maternal, fetal, and neonatal death, particularly in developing countries. Arsenic (As), which is commonly found in soil and groundwater, has been associated with various complications of pregnancy, such as spontaneous abortion, hypertension, and stillbirth. Hence, the study was used to explore the relationship between PE and blood concentration of As in this study. Blood concentration of As during pregnancy was measured by inductively coupled plasma mass spectrometry (ICP-MS). The results shown that the mean blood concentration of As was gradually increased from the control group to the severe PE group (P < 0.0001). Elevated blood concentration of As was associated with the prevalence of PE (OR = 12.81, 95% CI: 2.43-67.39 and 27.55, 1.75-433.43 for middle and high vs. low). Furthermore, elevated blood concentration of As was associated with the severity of PE. Additionally, we observed that blood concentration of As was associated with the hypoproteinemia (P = 0.001, rs = 0.37). Blood concentration of As was negatively corelated with the mean corpuscular volume (MCV) (P = 0.040, rs = - 0.23) and positively corelated with the mean corpuscular hemoglobin concentration (MCHC) (P = 0.044, rs = 0.23). Overall, our results indicated that the blood concentration of As can significantly predict the occurrence of PE. Additionally, we provided evidence that blood concentration of As may affect the occurrence of hypoproteinemia. These findings may provide some ideas for the prevention of PE and pregnancy complications.

Nanobead-Based Screening Method for Antibody Pairing of Dengue Virus Nonstructural Protein-1.

Dengue fever is a classic mosquito viral disease. Dengue virus non-structural protein-1 as a membrane-associated homologous dimer anchored to the surface of infected cells and also secreted into the blood. The nonstructural protein-1 levels are related to disease severity, and the presence of nonstructural protein-1 secreted from cells to the serum of people infected with the dengue virus is an early marker of infection. Paired antibodies are key in the establishment of rapid detection technology. In this study, the prepared recombinant nonstructural protein-1 protein of dengue virus serotype 3 was purified by the prokaryotic expression, and prepared monoclonal antibodies by cell fusion. A method for paired antibody screening was established based on the N-hydroxy succinimide-nanobeads and the prepared monoclonal antibodies. A simple and rapid point-of-care system integrating the paired antibodies and lateral flow assay was established to verify the screened antibody pairs. The results confirmed that the antibody pair screening method based on N-hydroxy succinimide-nanobeads is feasible.

Genome wide methylation analysis to uncover genes related to recurrent pregnancy loss.

BACKGROUND: Recurrent pregnancy loss (RPL) refers to two or more consecutive spontaneous abortion before 24 weeks of gestation, representing 1% of couples of childbearing age. Epigenetic factors including dysregulation of DNA methylation of some genes may play a role in RPL. OBJECTIVE: To identify RPL related genes modulated by DNA methylation expressed in decidua and blood. METHODS: Three decidua samples each from RPL patients and normal controls were recruited to perform genome-wide bisulfite sequencing (GWBS) and transcriptome sequencing. Based on the above results, 22.52 kb of differential methylation regions (DMRs) from 17 genes were verified by bisulfite sequencing PCR at specific region (Hi-MethylSeq) in another 15 decidua (7RPL vs. 8 Controls) and 13 blood (5RPL vs. 8 Controls) samples. RESULTS: 23 genes showed significantly differential cytosine methylation status and distinct expression level between PRL patients and healthy controls synergistically. Three signaling pathways were found to be shared between genes with both hypomethylated differential methylation regions (DMR) and upregulated differential gene expression (DGE). The results from Hi-MethylSeq showed that the hypermethylation of SGK1 in both blood and decidua samples in RPL patients, which was consistent to its lower expression in endometrium reported earlier. SGK3 and CREB5 also showed modulated methylation level in RPL decidua. CONCLUSION: Our finding supported that aberrant methylation of SGK1 and CREB5 could be a cause of the dysregulation of these gens in the endometrium, which is one of cause of reproductive failure. The function of SGK3 in reproduction system deserves further investigation.

[Preparation and application of anti-Mullerian hormone immunomagnetic beads].

Objective To prepare human anti-Mullerian hormone (AMH) immunomagnetic beads and HRP-labeled antibodies and establish a rapid double-antibody sandwich ELISA based on nanometer magnetic beads. Methods The expression vector of human AMH protein was constructed, and the recombinant AMH protein was expressed and purified. BALB/c mice were immunized with the recombinant protein to prepare the polyclonal antibody. Spleen cells were fused with myeloma Sp2/0 cells by PEG. Hybridoma cell lines which could stably secret monoclonal antibodies against AMH were screened out by ELISA. Monoclonal antibodies were produced from the ascites fluid of mice injected intraperitoneally with hybridoma cells and evaluated by Western blotting. Polyclonal antibodies purified from protein A were coupled to nano-magnetic' beads and used as capture antibodies, while HRP-labeled monoclonal antibody was prepared by sodium periodate method and used as probe antibody. A double antibody sandwich ELISA based on the nano-magnetic beads was established and optimized. Results A monoclonal antibody with good specificity for AMH was obtained,' and its subtype was IgG2b. The titers of purified polyclonal antibodies and monoclonal antibodies were up to 1:51 200. The capture antibody coupled with magnetic beads and the probe antibody labeled with HRP kept their good activity. The established method could detect AMH antigen within 1 hour and the detection limit was 50 ng/mL. Conclusion The prepared AMH immunomagnetic beads can be used for the fast and visualized detection of recombinant AMH.

[Clinical Significance of Screening Thalassemia with High- Throughput Sequencing].

OBJECTIVE: To evaluate the diagnostic value of high-throughput sequencing (NGS) in screening the thalassemia genes. METHODS: The peripheral blood of 2 858 cases of pre-pregnancy and pregnancy from October 2014 to October 2016 randomly were collected in department of obstetrics, the third people's hospital in Dongguan city. Peripheral blood was used for the blood routine examination, hemoglobin electrophoresis, traditional thalassaemia gene screening and NGS. RESULTS: The rate of missed diagnosis for α-thalassemia and ß-thalassemia using NGS was 0.87% and 1.59%, respectively. Meanwhile, the missed rate of screening for α-thalassemia and ß-thalassemia by traditional screening models was 26.77% and 2.38%, respectively. The area under the ROC curve of α-thalassemia and ß-thalassemia screened by NGS was 0.994 and 0.991, respectively, however, the area under the ROC curve of screening for α-thalassemia and ß-thalassemia by the traditional screening model was 0.866 and 0.988, respectively. The sensitivity, rate of missed diagnosis, Youden index and negative predictive value of screening for α-thalassemia and ß-thalassemia using NGS all were superior to those using traditional screening. CONCLUSION: Compared with the traditional screening model, the NGS screening for thalassemia genes shows a high accuracy, moreover can avoid missed diagnosis resulted from screening by conventional method, suggesting that the NCS possesses the accurate and diagnostic value for screening of thalassemia and can widely apply to clinical practise so as to provid the guarantee for early diagnosis of thalassemia.

Prenatal Diagnosis and Molecular Analysis of a Large Novel Deletion (- -JS) Causing α0-Thalassemia.

α-Thalassemia (α-thal) is a very common single gene hereditary disease caused by large deletions or point mutations of the α-globin gene cluster in tropical and subtropical regions of the world. Here, we report for the first time, a novel large α-thal deletion in a Chinese family from Jiangsu Province, People's Republic of China (PRC), which removes almost the entire α2 and α1 genes from the α-globin gene cluster. Thus, it was named the Jiangsu deletion (- -JS) on the α-globin gene cluster causing α0-thal. Heterozygotes for this deletion showed an α-thal trait phenotype with reduced mean corpuscular volume (MCV) and mean corpuscular hemoglobin (Hb) (MCH) levels. The sequencing results showed that a 2538 bp deletion (NG_000006.1: g.35801_38338) existed in this novel genotype on the basis of -α4.2 (leftward), indicating a deletion of about 6.8 kb from the α-globin cluster. In addition, a 29 bp sequence was inserted into the deletion during the recombination events that led to this deletion. Through pedigree analysis, we knew that the proband inherited the novel allele from his mother.

Physicochemical properties and in vitro antioxidant activities of polysaccharide from Artocarpus heterophyllus Lam. pulp.

A water-soluble polysaccharide from Artocarpus heterophyllus Lam. (jackfruit) pulp (JFP-Ps) was purified and its physicochemical properties were investigated. The in vitro antioxidant activities of JFP-Ps was evaluated by measuring DPPH and OH radicals scavenging activities, as well as reducing power. The results showed that JFP-Ps contained 79.12% of total sugar, 5.83% of protein, 15.65% of uronic acid, and 15 kinds of amino acids with high levels of Asp, Glu, Val, Leu and Lys. JFP-Ps was mainly composed of Rha, Ara, Gal, Glc, Xyl and GalA, with an average molecular weight of 1668kDa. FT-IR results showed the bands at the range of 1200-850cm-1 suggested the presence of carbohydrates in JFP-Ps. The results of antioxidant activities showed that JFP-Ps exhibited strong DPPH and OH radical scavenging activities, with a relatively lower reducing power, suggesting that JFP-Ps can be exploited as effective natural antioxidant applications in medical and food industries.