Analysis of the diagnostic performance of PAX1/SOX1 gene methylation in cervical precancerous lesions and its role in triage diagnosis.

Methylation panels, tools for investigating epigenetic changes associated with diseases like cancer, can identify DNA methylation patterns indicative of disease, providing diagnostic or prognostic insights. However, the application of methylation panels focusing on the sex-determining region Y-box 1 (SOX1) and paired box gene 1 (PAX1) genes for diagnosing cervical lesions is under-researched. This study aims to examine the diagnostic performance of PAX1/SOX1 gene methylation as a marker for cervical precancerous lesions and its potential application in triage diagnosis. From September 2022 to April 2023, 181 patients with abnormal HPV-DNA tests or cytological exam results requiring colposcopy were studied at Hubei Maternal and Child Health Hospital, China. Data were collected from colposcopy, cytology, HPV-DNA tests, and PAX1/SOX1 methylation detection. Patients were categorized as control, cervical intraepithelial neoplasia Grade 1 (CIN1), Grade 2 (CIN2), Grade 3 (CIN3), and cervical cancer (CC) groups based on histopathology. We performed HPV testing, liquid-based cytology, and PAX1/SOX1 gene methylation testing. We evaluated the diagnostic value of methylation detection in cervical cancer using DNA methylation positivity rate, sensitivity, specificity, and area under the curve (AUC), and explored its potential for triage diagnosis. PAX1/SOX1 methylation positivity rates were: control 17.1%, CIN1 22.5%, CIN2 100.0%, CIN3 90.0%, and CC 100.0%. The AUC values for PAX1 gene methylation detection in diagnosing CIN1+, CIN2+, and CIN3+ were 0.52 (95% confidence interval [CI]: 0.43-0.62), 0.88 (95% CI: 0.80-0.97), and 0.88 (95% CI: 0.75-1.00), respectively. Corresponding AUC values for SOX1 gene methylation detection were 0.47 (95% CI: 0.40-0.58), 0.80 (95% CI: 0.68-0.93), and 0.92 (95% CI: 0.811-1.00), respectively. In HPV16/18-negative patients, methylation detection showed sensitivity of 32.4% and specificity of 83.7% for CIN1+. For CIN2+ and CIN3+, sensitivity was all 100%, with specificities of 83.0% and 81.1%. Among the patients who underwent colposcopy examination, 166 cases had cytological examination results ≤ASCUS, of which 37 cases were positive for methylation, and the colposcopy referral rate was 22.29%. PAX1/SOX1 gene methylation detection exhibits strong diagnostic efficacy for cervical precancerous lesions and holds significant value in triage diagnosis.

Detection of DNA Methylation in Gene Loci ASTN1, DLX1, ITGA4, RXFP3, SOX17, and ZNF671 for Diagnosis of Cervical Cancer.

Objective: To evaluate the diagnostic value of DNA methylation detection of multiple gene loci in cervical cancer. Methods: A total of 61 cases requiring cervical biopsy were selected from the outpatient clinic of Maternal and Child Health Hospital of Hubei Province between January 2018 and December 2019. The patients were divided into four groups based on histopathologic diagnosis: cervical cancer (CC) group, high-grade squamous intraepithelial lesion (HSIL) group, low-grade squamous intraepithelial lesion (LSIL) group, and control group. HPV examination, liquid-based cytology examination, and DNA methylation detection at multiple gene sites were performed. The positive rate of DNA methylation, sensitivity, specificity, area under the curve (AUC), and other efficacy indexes were calculated to evaluate the diagnostic value of DNA methylation detection at multiple gene loci in cervical cancer. Results: The positive rates of DNA methylation in CC, HSIL, LSIL, and control groups were 100%, 88%, 83% and 17%, respectively. The ZNF671 gene had the highest positive rate among the cervical lesion group, with rates of 57%, 76%, and 100% in LSIL, HSIL, and CC groups respectively. The combination of DNA methylation detection at multiple gene loci showed the highest diagnostic efficacy for HSIL and cervical cancer, with AUC value of 0.850 (95% CI:0.746-0.954), a Youden index of 0.654, and a sensitivity and specificity of 85% and 85.4%, respectively. The diagnostic efficacy of the combined detection was significantly higher than that of HPV examination and liquid-based cytology examination (P < 0.05). Conclusion: DNA methylation detection at multiple gene loci is highly effective and diagnostic tool for cervical cancer, and has potential application value in clinical practice.

Effect of tannic acid-OSA starch complexation on the binding capacity and release of aldehydes off-flavor in aqueous matrix.

In order to further clarify the regulation of tannic acid on the off-flavor in starch-based algal oil emulsions, the effect of different starch matrix (OSA starch and OSA starch-tannic acid complex) on the release capacities of aldehydes (pentanal, hexanal, heptanal, nonanal) were investigated. The adsorption and retention ability, thermodynamic parameters, and hydrophobicity of aldehydes in the starch matrix were analyzed. Nonanal exhibited the strongest adsorption ability (65.01%-85.69%) with the starch matrix, followed by heptanal, hexanal, and pentanal, which accounted for the structures of aldehydes. Furthermore, aldehydes had a higher affinity with complex (16.33%-83.67%) than OSA starch (9.70%-66.71%) because the tannic acid altered the structure of OSA starch. Isothermal titration calorimetry suggested that the interaction between the starch matrix and aldehydes was an entropy-driven spontaneous endothermic reaction, and hydrophobic interactions were the predominant driving forces. Altogether, these results lay a theoretical foundation for facilitating the regulation of flavor in starch foods.

First Report of Leaf Spot Caused by Lasiodiplodia theobromae on Kadsura coccinea in China.

Kadsura coccinea (Lem.) A. C. Smith is an evergreen liana widely cultivated in China for its economic importance in traditional medicine. Many phytochemical studies on the stems and roots of K. coccinea have shown a variety of biological activities, such as anti-hepatitis, anti-HIV, and anti-tumor (Yang et al. 2020). In July 2021, symptoms of leaf spot were observed in a plantation of K. coccinea in Longan (23°03´N, 107°54´E), Guangxi province, China. The incidence of this disease was 36%, and severity varies from approximately 20 to 40% of leaf surface coverage. Symptoms began as small brown spots that expanded into irregular to nearly flower-shaped lesions. To isolate the pathogen, leaves with spots were collected, sterilized with 75% ethanol for 15 s followed by 2% sodium hypochlorite for 120 s, rinsed three times in sterilized distilled water, cut into 5 × 5 mm pieces, and placed onto potato dextrose agar (PDA) plates. The plates were kept in an incubator at 26°C in the dark for at least 2 days. A total of 27 fungal colonies of similar morphology out of 30 pieces of infected tissues were isolated. Four representative isolates (HBB1 to HBB4) were selected to study for further characterization. Fungal colonies were initially grayish-white and then turned greenish-gray on PDA. The black pycnidium and immature conidia appeared over PDA plates after 18 days. The immature conidia were colorless and transparent, elliptical, and had a single-cell structure. After 5 days, the immature conidia gradually become black and develop into mature conidia. The mature conidia were dark brown and two-celled with longitudinal striations, 20.41-29.93 × 12.42-17.19 µm (average 26.07×14.51 µm; n = 100). For DNA-based identification, the internal transcribed spacer (ITS) region, translation elongation factor 1 alpha (EF1-α), and ß-tubulin (TUB) genes of the isolates were amplified and sequenced using the primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. Sequences were submitted to GenBank (Accession nos. MW045412 to MW045415 for ITS, MW065559 to MW065562 for EF1-α, and MW065555 to MW065558 for TUB). A phylogenetic analysis was conducted using the Maximum Likelihood method on concatenated sequences of the three genes, which showed that the four Chinese isolates from K. coccinea were clustered with reference isolates of Lasiodiplodia theobromae including the ex-neotype CBS 164.96. Pathogenicity tests were performed on young, fully expanded leaves of 2-year seedlings. A 10 µL conidial suspension (1×106 conidia/mL) was inoculated on each wound on the left-half leaf and a 10 µL sterile water was inoculated on each wound on the right-half leaf (control). Each treatment was repeated three times. Inoculated leaves were wrapped in plastic bags for 5 days and plants were maintained in a growth chamber at 27°C, 85% relative humidity. Brown leaf spots appeared 5 to 6 days after inoculation, whereas the control leaves treated with sterile water showed no symptoms. All re-isolations from spots produced colonies with the same morphological characters as L. theobromae, completing Koch's postulates. To our knowledge, this is the first report of L. theobromae causing leaf spot on K. coccinea in China and worldwide. Severe leaf disease caused by L. theobromae threatens K. coccinea production. The disease threatens K. coccinea growth, and effective control measures should be identified to reduce losses.

Liposomes for encapsulation of liposoluble vitamins (A, D, E and K): Comparation of loading ability, storage stability and bilayer dynamics.

To understand the encapsulation difference and stability mechanism of nanoliposomes (NLPs) loaded with different kinds and loads of liposoluble vitamins (LSV, including VA, VD, VE, and VK), the physicochemical stability during three-months storage and bilayer membrane properties of LSV-NLPs were evaluated. The results suggested that VD and VE were not suitable for high-load (≥30 wt%) encapsulation, but the stability of other LSV-NLPs was excellent during storage. Their particle size was less than 100 nm, the polydispersity index was less than 0.3, and the retention rate of VE and VK remained above 85 %. LSV encapsulation inhibited malondialdehyde production, decreased liposome surface roughness, and improved nanoliposome rigidity. The order of occupying capacity of LSV to the hydrophobic zone of the bilayer was VK＞VD＞VE＞VA, and the stability of LSV located in the hydrophobic region was better. Except for high-load VD and VE, the other LSV encapsulation increased the microviscosity of the lipid-water interface and hydrophobic zone by 0.5 âˆ¼ 7.1 times and 0.5 âˆ¼ 20 times, respectively. The accumulation of acyl chain was enhanced by 0.2 âˆ¼ 4 times, and the interchain longitudinal and intra-chain transverse order degree was increased by 10.89 %∼144.35 % and 3.26 %∼115.52 %, respectively. High microviscosity and tight chain stacking limited bilayer fluidity and thus improve LSV-NLPs stability. This work will contribute to the application of nanoliposomes as liposoluble vitamin carriers in the food industry.

The selective encapsulation and stabilization of cinnamaldehyde and eugenol in high internal phase Pickering emulsions: Regulating the interfacial properties.

This work aimed to investigate the encapsulation and stabilization mechanism of cinnamaldehyde and eugenol in high internal phase Pickering emulsions (HIPPEs) through regulating their interfacial rheological properties and interfacial microstructure. With the incorporation of cinnamaldehyde, the Schiff base reaction between the cinnamaldehyde and proteins favored the formation of the predominantly elastic and solid-like interfacial layers. In contrast, the hydrogen bonds between eugenol and proteins resulted in the transformation of interfacial layers to viscous dominant with weak viscoelastic responses. Thus, cinnamaldehyde-loaded HIPPEs had a better storage stability than eugenol-loaded HIPPEs, and the retention rate was increased by about 15 %∼20 %. The addition of tea camellia seed oil inhibited the mobility of immobilized water and improved the retention rates of cinnamaldehyde and eugenol by approximately 6 % and 12 % (30 days at 25 °C), respectively. These findings will be beneficial for the development and design of effective essential oil encapsulation systems in the food industry.

First Report of Leaf Spot Caused by Phyllosticta capitalensis on Illicium difengpi in China.

Illicium difengpi B. N. Chang et al., a shrub with aromatic odor in the Illicium genus, is extensively used as a medicinal plant in China. In June of 2020, a leaf spot on I. difengpi with incidence of about sixty percent was observed in a field located in Guilin (25°4'40"N; 110°18'21"E), Guangxi Province, China. Initial leaf symptoms were round spots with gray centers, surrounded by yellow halos. The spots gradually spread and merged. Six samples of symptomatic leaves were collected from six diseased plants, and they were surface disinfested before isolation. Potato dextrose agar (PDA) was used to culture pathogens. Successively, pure cultures were obtained by transferring hyphal tips to new PDA plates. A total of 10 isolates were obtained from the affected leaves. Two single-spore isolates (GX-1 and GX-2) were obtained and confirmed to be identical based on morphological characteristics. The representative isolate GX-2 was selected for further study on morphological and molecular characteristics. The colony of isolate GX-2 was about 4 cm in diameter on a PDA plate in 5 days, dark green with a granular surface, and irregular white edge. Conidia were hyaline, unicellular, oval, narrow at the end with a single apical appendage, and 8.2 to 13.8 × 3.7 to 7.2 µm (n = 50). Spermatia were hyaline, bacilliform with swollen ends, 3.8 to 8.9 × 1.3 to 1.9 µm (n = 50). Morphological characteristics of isolate GX-2 were consistent with the description of Phyllosticta capitalensis (Wikee et al. 2013). The internal transcribed spacer (ITS) region, translation elongation factor 1-α (tef1-α), glyceraldehyde-3-phosphate dehydrogenase (GPDH) and actin (ACT) were amplified using primers ITS1/ITS4, EF-728F/EF-986R, Gpd1-LM/Gpd2-LM and ACT-512F/ACT-783R, respectively (Wikee et al. 2013). Sequences were deposited in GenBank with accession numbers OL505439 for ITS, OL539429 for ACT, OL539430 for tef1-α and OL539431 for GPDH. BLAST analysis in GenBank showed that these sequences were 99 to 100% similar to the corresponding ITS (MT649668), ACT (MN958710), tef1-α (MN958711) and GPDH (KU716077) sequences of P. capitalensis. Also, the phylogenetic tree based on genes of ITS, tef1-α, GPDH and ACT by the maximum likelihood method showed that isolate GX-2 clustered together with P. capitalensis. The pathogenicity tests were carried out on a healthy 3 year-old plant in the greenhouse with 80% relative humidity at 25 °C. Four sterilized leaves were wounded with a needle and inoculated with 20 µL spore suspension (1 × 106 spores/ml). Another four sterilized leaves were inoculated with 20 µL sterile water as a control. All plants were incubated in a chamber with 98% relative humidity at 25 ± 1°C. After 12 days, disease symptoms similar to the field were observed on leaves, whereas control plants remained healthy. P. capitalensis was successfully reisolated only from the inoculated leaves and identified based on morphological characters. P. capitalensis caused leaf spots on various host plants around the world (Wikee et al. 2013), including on tea plants in China (Cheng et al. 2019) and oil palm in Malaysia (Nasehi et al. 2020), but it has not been reported on I. difengpi. Thus, this is the first report of P. capitalensis causing leaf spot on I. difengpi. This study will provide an important reference for the control of the disease. The epidemiology of this disease should be investigated in further research.

Complete genome sequence of biocontrol strain Paenibacillus peoriae HJ-2 and further analysis of its biocontrol mechanism.

BACKGROUND: Paris polyphylla is a herb widely used in traditional Chinese medicine to treat various diseases. Stem rot diseases seriously affected the yield of P. polyphylla in subtropical areas of China. Therefore, cost-effective, chemical-free, eco-friendly strategies to control stem rot on P. polyphylla are valuable and urgently needed. RESULTS: In this paper, we reported the biocontrol efficiency of Paenibacillus peoriae HJ-2 and its complete genome sequence. Strain HJ-2 could serve as a potential biocontrol agent against stem rot on P. polyphylla in the greenhouse and field. The genome of HJ-2 consists of a single 6,001,192 bp chromosome with an average GC content of 45% and 5,237 predicted protein coding genes, 39 rRNAs and 108 tRNAs. The phylogenetic tree indicated that HJ-2 is most closely related to P. peoriae IBSD35. Functional analysis of genome revealed numerous genes/gene clusters involved in plant colonization, biofilm formation, plant growth promotion, antibiotic and resistance inducers synthesis. Moreover, metabolic pathways that potentially contribute to biocontrol mechanisms were identified. CONCLUSIONS: This study revealed that P. peoriae HJ-2 could serve as a potential BCA against stem rot on P. polyphylla. Based on genome analysis, the genome of HJ-2 contains more than 70 genes and 12 putative gene clusters related to secondary metabolites, which have previously been described as being involved in chemotaxis motility, biofilm formation, growth promotion, antifungal activity and resistance inducers biosynthesis. Compared with other strains, variation in the genes/gene clusters may lead to different antimicrobial spectra and biocontrol efficacies.

First Report of Anthracnose on Kadsura coccinea Caused by Colletotrichum siamense in China.

Kadsura coccinea (Lem.) A. C. Smith, an evergreen liana, is widely cultivated in China for its economic importance in traditional medicine. Many phytochemical studies on the stems and roots of K. coccinea have shown numerous biological activities, such as anti-tumor, anti-HIV, and anti-oxidant (Yang et al. 2020). In June 2019, an anthracnose on K. coccinea was observed in a plantation in Longan (23°03´N, 107°54´E), Guangxi province. Disease incidence was up to 30% in a plantation. Its symptoms began as small brown spots that expanded into nearly circular spots (Fig. 1A). To isolate pathogen, diseased leaves were collected. The leaves were sterilized with 75% ethanol for 15 s followed by 2% sodium hypochlorite for 90 s, then rinsed three times in sterilized distilled water, cut into 5 × 5 mm pieces, and placed into potato dextrose agar (PDA) plates. The plates were incubated in an incubator at 25°C in dark for 2-3 days. Fungal colonies with similar morphology of 27 isolates were isolated from the 30 infected tissues. Six representative isolates (YB1 to YB6) were selected to further study their characterization. Fungal colonies were grayish-white, orange-yellow conidial masses could be observed in colonies (Fig. 1C). The mature conidia were colorless and transparent, elliptical, and single-celled, 13.0-21.0 × 4.0-8.0 µm (average 16.92 × 5.92 µm; n =100) (Fig. 1B). The DNA sequences of ribosomal internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate (GAPDH), calmodulin (CAL), actin (ACT), chitin synthase (CHS-1) and ß-tubulin (TUB2) were amplified by PCR using the primer pairs ITS1/ITS4, GDF/GDR, CL1C/CL2C, ACT-512F/ACT-783R, CHS-79F/CHS-354R, and T1/Bt2b (Wang et al. 2020), respectively. Sequences were submitted to GenBank (Accession nos. MZ040489 to MZ040494 for ITS, MZ069043 to MZ069048 for GAPDH, MZ069049 to MZ069054 for CAL, MZ069055 to MZ069060 for ACT, MZ069061 to MZ069066 for CHS-1, and MZ069067 to MZ069072 for TUB2). These sequences were 98%-100% identical to that of reference isolates JX010278, JX010019, JX009709, GQ856775, GQ856730, and JX010410 of Colletotrichum siamense CBS 125378 ex-type recorded in GenBank. Phylogenetic analysis of combined ITS, GAPDH, CAL, ACT, CHS-1, TUB2 genes with 16 sequences obtained from GenBank using maximum likelihood method showed that the six isolates clustered with two reference isolates of Colletotrichum siamense as a distinct clade (Fig. 2). Based on morphological characteristics and phylogenetic analysis, six isolates were identified as C. siamense. Pathogenicity tests were performed on young, fully expanded leaves of 1-year seedlings. Every leaf was punctured at 6 points on the right half and 6 points on the left half using a sterile needle. A 10 µl conidial suspension (1×106 conidia/ml) was inoculated on each wound on the left-half leaf and a 10 µl sterile water was inoculated on each wound on the right-half leaf (control). Each treatment was repeated three times. Inoculated leaves were wrapped in plastic bags for 2 days and after removing the bags, plants were maintained in a growth chamber at 28°C, 80% relative humidity, and a 12-h photoperiod. Anthracnose spots formed 2 to 3 days after inoculation, whereas the control leaves remained symptomless. Morphological characters matched the descriptions of C. siamense. The pathogen was previously reported to cause anthracnose on Aloe vera (Azad et al. 2020), postharvest anthracnose in mango (Liu et al. 2017), pod rot in cacao (Serrato-Diaz et al. 2020). To our knowledge, this is the first report of anthracnose on K. coccinea caused by C. siamense in China.

Enhanced nitrogen and phosphorus removal by natural pyrite-based constructed wetland with intermittent aeration.

Four subsurface flow constructed wetlands (SFCWs) filled with different substrates including ceramsite, ceramsite+pyrite, ceramsite+ferrous sulfide, and ceramsite+pyrite+ferrous sulfide (labeled as SFCW-S1, SFCW-S2, SFCW-S3, and SFCW-S4) were constructed, and the removal of nitrogen and phosphorus by these SFCWs coupled with intermittent aeration in the front section was discussed. The key findings from different substrate analyses, including nitrification and denitrification rate, enzyme activity, microbial community structure, and the X-ray diffraction, revealed the nitrogen and phosphorus removal mechanism. The results showed that the nitrogen and phosphorus removal efficiency for SFCW-S1 always remained the lowest, and the phosphorus removal efficiency for SFCW-S4 was recorded as the highest one. However, after controlling the dissolved oxygen by intermittent aeration in the front section of SFCWs, the nitrogen and phosphorus removal efficiencies of SFCWs-S2 and S4 became higher than those of SFCW-S1, and SFCW-S3. It was noticed that the pollutants were removed mainly in the front section of the SFCWs. Both precipitation and adsorption on the substrate were the main mechanisms for phosphorus removal. A minute difference of nitrification rate and ammonia monooxygenase activity was observed in the SFCWs' aeration zone. The denitrification rates, nitrate reductase, nitrite reductase, and electron transport system activity for SFCW-S2 and SFCW-S4 were higher than those detected for SFCW-S1 and SFCW-S3 in the non-aerated zone. Proteobacteria was the largest phyla found in the SFCWs. Moreover, Thiobacillus occupied a large proportion found in SFCW-S2, and SFCW-S4, and it played a crucial role in pyrite-driven autotrophic denitrification.

Migration kinetic of silver from polylactic acid nanocomposite film into acidic food simulant after different high-pressure food processing.

The migration study of nano-Ag migration from polylactic acid (PLA) films was studied. Samples treated by high-pressure food processing (0, 100, 200, 300, and 400 MPa pressure) were soaked in acetic acid solution and incubated at 20 °C for 90 days. At the end of storage, nano-Ag particles (AgNPs) migration from the PLA/AgNPs composite film treated under 200 MPa high pressure was the lowest. However, AgNPs migration was accelerated under 400 MPa high pressure. High-pressure processing (200 MPa) could cause denser structure and higher crystallinity degree in films than other treatments. Lower amount of AgNPs induced a decline in the intensity of specific characteristic peaks. The diffraction peak intensity of α-crystal for the film sample treated with 400 MPa was the lowest on day 60. The crystallization index of the PLA matrix changed with different high-pressure processing. The result indicated that appropriate high-pressure food processing could effectively suppress AgNPs migration from PLA-based film while contacting with acidic acid food simulant. PRACTICAL APPLICATION: The release of nanoparticles from food packaging material is a very important matter when the migration is concerned with regulatory and toxicity issues. The study described the migration kinetic of AgNPs from PLA nanocomposite film into acidic food simulant after different high-pressure food processing. The results indicated that the PLA/AgNPs nanocomposite film was safe for acidic food after high-pressure treatment.

Effect of ultrahigh-pressure treatment on the functional properties of poly(lactic acid)/ZnO nanocomposite food packaging film.

BACKGROUND: Our living environment is being increasingly polluted by petroleum-based plastics and there is an increasing demand for biodegradable food packaging. In this study, the effect of various ultrahigh-pressure (UHP) treatments (0, 200 and 400 MPa) on the microstructure and thermal, barrier and mechanical properties of poly(lactic acid) (PLA)/ZnO nanocomposite films was studied. RESULTS: The film-forming solution was processed using UHP technology. The crystallinity, strength and stiffness of the composite film after UHP treatment increased. In addition, barrier property analysis showed that the UHP treatment significantly (P < 0.05) reduced the oxygen permeability and water vapor permeability (WVP) coefficient of the PLA/ZnO nanocomposite film. Furthermore, the WVP value of the film treated at 400 MPa (50 g kg-1 nano-ZnO content) was the lowest and reduced by 47.3% compared with that of pure PLA film. The improvement in these properties might be due to the interaction between nano-ZnO and PLA matrix promoted by UHP treatment. CONCLUSIONS: Therefore, the application of UHP technology on the film-forming solution could improve the crystallinity and functional properties of the nanocomposite film, and has great potential in the production of food packaging films with ideal functions. © 2021 Society of Chemical Industry.

Pregnancy complications among nulliparous and multiparous women with advanced maternal age: a community-based prospective cohort study in China.

BACKGROUND: This study aimed to evaluate the incidence rates and risks of pregnancy complications among nulliparous and multiparous women with advanced maternal age (AMA, ≥35 years) in China. METHODS: We performed a community-based prospective cohort study of 10,171 pregnant women in selected two sub-districts and 11 towns of Liuyang from 2013 to 2015. All subjects were followed up from the first prenatal care (at ≤12 weeks) to delivery, and risks of pregnancy complications were compared by parity and maternal age groups. RESULTS: Among nulliparas, women with AMA showed significantly increased risks for gestational hypertension (OR 8.44, 95%CI 1.68-2.88), preeclampsia/eclampsia (OR 9.92, 95%CI 4.87-18.78), premature rupture of membrane (OR 6.84, 95%CI 2.00-17.69), as compared to women in the 20-29-year age group. Among multiparas with AMA, increased risks were found for gestational diabetes mellitus (OR 3.29, 95%CI 1.76-5.94), anemia (OR 1.85, 95%CI 1.25-2.69), polyhydramnios (OR 3.29, 95%CI 1.56-6.64), premature rupture of membrane (OR 5.14, 95%CI 2.12-12.29), and preterm labor (OR 1.89, 95CI 1.42-2.50). CONCLUSIONS: Women with AMA were associated with increased risks of pregnancy complications, and complications with increased risks differed in nulliparas and multiparas. Women with AMA should be identified as a high-risk group in clinical practice.

Barrier Properties and Characterizations of Poly(lactic Acid)/ZnO Nanocomposites.

This study aimed to reinforce the barrier performance (i.e., oxygen-gas and water-vapor permeability) of poly(lactic acid) (PLA)-based films. Acetyltributylcitrate and zinc oxide nanoparticle (nano-ZnO), serving as plasticizer and nanofiller, respectively, were blended into a PLA matrix through a solvent-volatilizing method. The structural, morphological, thermal, and mechanical performances were then studied. Scanning electron microscopic images showed a significant dispersion of nano-ZnO in PLA ternary systems with low nano-ZnO content. The interaction between PLA matrix and ZnO nanoparticles was further analyzed by Fourier-transform infrared spectroscopy. Wide-angle X-ray scattering spectroscopy demonstrated high compatibility between PLA matrix and ZnO nanoparticles. Mechanical property studies revealed good tensile strength and low flexibility. Differential scanning calorimetry curves proved that an amorphous structure mostly existed in PLA ternary systems. The improvements in barrier property and tensile strength indicated that the PLA/nano-ZnO composite films could be used for food packaging application.

The Performance Changes and Migration Behavior of PLA/Nano-TiO2 Composite Film by High-Pressure Treatment in Ethanol Solution.

To study the relationship between performance changes and nanoparticles migration of the composite film at different migration stages, the poly (lactic acid) (PLA)/nano-TiO2 composite film treated by high pressure was immersed in 50% (v/v) ethanol solution for 45 days at 40 °C, and the film characteristics and migration behavior were analyzed. The results showed that the migration of the composite film with the highest loading of nano-TiO2 (20 wt. %) in alcoholic food simulated solution was far less than 10 mg/kg during the 45-day migration process. Although with the increase of migration time, the micro-morphology of composite film became rougher, the crystallinity decreased and the gas permeability increased, but the internal crystal structure of the composite film remained basically unchanged. The PLA/nano-TiO2 composite films treated by high pressure treatment were relatively stable, and had good performance and migration behavior in alcoholic food simulated solution, the nanocomposite film after high pressure treatment could be used to reduce nano-TiO2 particle migration and subsequently reduce human exposure as the packaging film for the packaging of alcoholic food, which provide a theoretical basis for the applications of high pressure treatment of PLA/nano-TiO2 composite films in food packaging material and broaden its application prospects.

Effect of High Pressure Treatment on Poly(lactic acid)/Nano⁻TiO2 Composite Films.

The microstructure, thermal properties, mechanical properties and oxygen and water vapor barrier properties of a poly(lactic acid) (PLA)/nano-TiO2 composite film before and after high pressure treatment were studied. Structural analysis showed that the functional group structure of the high pressure treated composite film did not change. It was found that the high pressure treatment did not form new chemical bonds between the nanoparticles and the PLA. The micro-section of the composite film after high pressure treatment became very rough, and the structure was depressed. Through the analysis of thermal and mechanical properties, high pressure treatment can not only increase the strength and stiffness of the composite film, but also increase the crystallinity of the composite film. Through the analysis of barrier properties, it is found that the barrier properties of composite films after high pressure treatment were been improved by the applied high pressure treatment.

Distinctive expression signatures of serum microRNAs in ischaemic stroke and transient ischaemic attack patients.

Circulating microRNAs (miRNAs) have recently emerged as promising biomarkers for ischaemic stroke (IS). However, the expression patterns of specific miRNAs in transient ischaemic attack (TIA) patients have not been investigated. Their predictive values for the presence of IS and TIA and their relationships to the neurological deficit severity of IS and the subsequent stroke risk after TIA remain unclear exactly. In this study, 754 miRNAs were initially screened by the TaqMan Low Density Array (TLDA) in two pooled serum samples from 50 IS patients and 50 controls. Markedly altered miRNAs were subsequently validated by individual quantitative reverse-transcription PCR (qRT-PCR) assays first in the same cohort of TLDA and further confirmed in another larger cohort including 177 IS, 81 TIA patients and 42 controls. Consequently, TLDA screening showed that 71 miRNAs were up-regulated and 49 miRNAs were down-regulated in IS patients. QRT-PCR validation confirmed that serum levels of miR-23b-3p, miR-29b-3p, miR-181a-5p and miR-21-5p were significantly increased in IS patients. Strikingly, serum levels of miR-23b-3p, miR-29b-3p and miR-181a-5p were also significantly elevated in TIA patients. Furthermore, up-regulated miR-23b-3p, miR-29b-3p and miR-21-5p could clearly differentiate between IS and TIA patients. Logistic regression and receiver-operating characteristic curve analyses demonstrated that these altered miRNAs may function as predictive and discriminative biomarkers for IS and TIA, and their distinctive expression signatures may contribute to assessing neurological deficit severity of IS and subsequent stroke risk after TIA.

[Serum non-esterified fatty acids to albumin ratio increased significantly in children with nephrotic syndrome].

OBJECTIVE: To analyze serum levels of non-esterified fatty acids (NEFA) and albumin (ALB) in children with nephrotic syndrome (NS) and investigate the clinical significance of altered serum NEFA to ALB ratio in children with NS in acute and remission phases. METHODS: Serum levels of NEFA and ALB were measured in 55 NS children in acute phase, in 33 NS children in remission and in 122 healthy control children, and the ratio of NEFA to ALB was calculated. The other lipid/lipoprotein and renal function parameters were also analyzed in these children. RESULTS: Compared with the healthy control children, children with NS had a significantly decreased serum ALB level (t=11.152, P<0.001) and a significantly increased NEFA to ALB ratio (t=4.991, P<0.001). Compared with NS children in remission, those in acute phase showed a significantly decreased ALB (Z=7.822, P<0.001) and an increased NEFA to ALB ratio (t=4.991, P<0.001). In all the NS children, NEFA to ALB ratio was positively correlated with the levels of TC (r=0.564, P<0.001), TG (r=0.444, P<0.001), LDL-C (r=0.625, P<0.001), urea (r=0.437, P<0.001), creatinine (r=0.278, P=0.013), and uric acid (r=0.397, P<0.001), while negatively correlated with the level of total protein (r=-0.461, P<0.001). Multiple linear regression analyses showed that NEFA to ALB ratio was independently associated with serum urea levels (ß=0.703, P=0.001; adjusted R2=0.494) after adjustment of other related factors. CONCLUSION: Serum NEFA to ALB ratio is significantly increased in NS children in close association with impaired kidney function, and may function as a novel parameter for assessing the development of NS.

Elevated serum miR-93, miR-191, and miR-499 are noninvasive biomarkers for the presence and progression of traumatic brain injury.

The levels of miR-93, miR-191, and miR-499 have been reported to be up-regulated in the tissues of experimental traumatic brain injury (TBI) rat models. However, the clinical diagnostic and prognostic values of the serum signatures of these 3 miRNAs in TBI remain unclear. The purpose of this study was to determine the expression levels of these 3 microRNAs (miRNAs) in the sera of TBI patients and to evaluate their relationships with the severity and clinical outcome of TBI. The serum levels of these miRNAs were assessed in TBI patients (n = 76) and healthy controls (n = 38) by quantitative reverse-transcription PCR. The severities and clinical outcomes of the TBI patients were evaluated with the Glasgow coma scale and the Glasgow outcome scale. The serum miR-93, miR-191, and miR-499 levels were significantly increased in the TBI patients compared with the controls at all examined time points, and these levels were significantly higher in the patients with severe TBI than in those with moderate or mild TBI (p < 0.05). The serum miR-93, miR-191, and miR-499 levels were significantly higher in the patients with a poor outcome than in those with a good outcome (p < 0.05). The AUCs of miR-93, miR-191, and miR-499 for distinguishing the TBI patients from the healthy controls were 1.000 (p < 0.001), 0.727 (p < 0.001) and 0.801 (p < 0.001), respectively. Interestingly, the AUCs of miR-93, miR-191, and miR-499 for distinguishing the mild TBI patients from the healthy controls were 1.000 (p < 0.001), 0.742 (p < 0.001) and 0.819 (p < 0.001), respectively. Taken together, these results indicate that miR-93, miR-191, and miR-499 are potentially valuable indicators of the diagnosis, severity, and prognosis of TBI. Our study showed that the serum levels of miR-93, miR-191, and miR-499 are all increased in traumatic brain injury (TBI) patients. Their serum levels are associated with TBI severity and outcome, which suggest that these miRNAs play important roles in the pathogenesis and progression of TBI. We think these findings should provide a new strategy for the diagnostic, prognostic, and treatment of TBI.

[Local heating of murine skin by millimeter waves based on HBHE].

The authors deduced Gaussian function of millimeter wave power distribution, and built up a transient thermal multilayer model for the heating of murine skin by high power millimeter waves with finite volume method (FVM) based on HBHE in the present paper. We analyzed the calculated results and compared them with the results calculated by Pennes' equation and the experimental ones; found that the temperature calculated by HBHE was more reasonable. Especially under high power millimeter wave, the calculated results were basically consistent with the experimental ones, and the superiority of the theoretical model was confirmed.