Molecular findings in patients for whole exome sequencing and mitochondrial genome assessment.

OBJECTIVE: Whole exome sequencing (WES) is becoming more widely used as a diagnostic tool in the field of medicine. In this article, we reported the diagnostic yield of WES and mitochondrial genome assessment in 2226 consecutive cases in a single clinical laboratory. MATERIALS AND METHODS: We retrospectively analyzed consecutive WES reports from 2226 patients with various genetic disorders. WES-process was focused exclusively on the probands and aimed at a higher diagnostic capacity. We determined the diagnostic rate of WES overall and by phenotypic category, mode of inheritance, mitochondrial genome variant, and copy number variants (CNVs). RESULTS: Among the 2226 patients who had diagnostic WES proband-only, the overall diagnostic yield of WES was 34.59% (770/2226). The highest diagnostic yield was observed in autosomal dominant disorders, at 45.58% (351/770), followed by autosomal recessive at 31.95%(246/770), X-linked disorder at 9.61%(74/770), and mitochondrial diseases at a notably lower 0.65%(5/770). The 12.21% (94/770) diagnoses were based on a total of 94 copy number variants reported from WES data. CNVs in children accounted for 67.02% of the total CNVs. While majority of the molecular diagnoses were related to nuclear genes, the inclusion of mitochondrial genome sequencing in the WES test contributed to five diagnoses. all mitochondrial diseases were identified in adults. CONCLUSIONS: The proband-only WES provided a definitive molecular diagnosis for 34.59% of a large cohort of patients while analysis of WES simultaneously analyzed the SNVs, exons, mitochondrial genome, and CNVs, thereby improving the diagnostic yield significantly compared to the single-detection WES method; and facilitating the identification of novel candidate genes.

[Genetic analysis of a case with Al Kaissi syndrome and a literature review].

OBJECTIVE: To explore the genetic etiology of a child with delayed growth and development and carry out a literature review. METHODS: A child suspected for Al Kaissi syndrome at the First Affiliated Hospital of Zhengzhou University on March 6, 2021 was selected as the study subject. Following extraction of genomic DNA, the child was subjected to copy number variation sequencing (CNV-seq) and whole exome sequencing (WES), and candidate variants were verified by PCR-agarose gel electrophoresis and quantitative real-time PCR (qPCR). Prenatal diagnosis was conducted on chorionic villi sample upon subsequent pregnancy. RESULTS: The child, a 6-year-and-4-month-old boy, has dysmorphic features including low-set protruding ears and triangular face, delayed language and intellectual development, and ventricular septal defect. CNV-seq result has found no obvious abnormality, whilst WES revealed homozygous deletion of exons 1 and 2 of the CDK10 gene, which was confirmed by PCR-agarose gel electrophoresis and qPCR. Both of his parents were heterozygous carriers. Prenatal diagnosis using chorionic villi samples suggested that the fetus also carried the heterozygous deletion. CONCLUSION: The clinical features of Al Kaissi syndrome in this child can probably be attributed to the homozygous deletion of exons 1 and 2 of the CDK10 gene.

Advances in emulsion-based delivery systems for nutraceuticals: Utilization of interfacial engineering approaches to control bioavailability.

Designing functional foods fortified with nutraceuticals is an important focus of modern food science with the aim of improving human health and wellbeing. However, many nutraceuticals have a low water solubility and poor physiochemical stability, which makes it challenging to incorporate into food matrices. Moreover, nutraceuticals may also have a low bioavailability after oral administration because they can either precipitate or chemically degrade, and/or might not be absorbed in the gastrointestinal tract. Numerous strategies have been developed and applied to encapsulate and deliver nutraceuticals. Emulsions are a kind of colloid delivery system where one phase is dispersed into another immiscible phase in the form of small droplets. These droplets have been widely used as carriers to improve the dispersibility, stability, and absorption of nutraceuticals. Many factors affect the formation and stability of emulsions, with the interfacial coating formed around the droplets by emulsifiers and other stabilizers being one of the most important. Hence, interfacial engineering principles are needed for the design and development of emulsions. Different approaches to interfacial engineering have been developed, which can help to modulate the dispersibility, stability, and bioavailability of nutraceuticals. This chapter summarizes recent research in developing interfacial engineering approaches and their impacts on the bioavailability of nutraceuticals.

Computable structured aptamer for targeted treatment of ovarian cancer.

Nucleolin protein expression is higher on the ovarian cancer cell surface. AS1411, a DNA aptamer, can bind with nucleolin protein specifically. In this study, we developed HA and ST DNA tiles to assemble six AS1411 aptamers to deliver doxorubicin. In addition, to superior serum stability and drug loading, HA-6AS and ST-6AS outperformed TDN-AS in cellular uptake. HA-6AS and ST-6AS exhibited satisfactory targeted cytotoxicity and achieved resounding lysosomal escape. Moreover, when injected into nude mice subcutaneous xenograft models, HA-6AS reached the peak in tumor more quickly than ST-6AS, and better expressed the active targeting ability of AS1411. Our study suggests that designing appropriate DNA tiles to assemble different aptamers to deliver different chemotherapeutic drugs is a promising treatment for ovarian cancer.

Rapid microRNA detection method based on DNA strand displacement for ovarian cancer cells.

The current cancer detection methods are heavily dependent on the component analysis of corresponding cancer antigens. There is a lack of effective and simple clinical methods of ovarian cancer screening, which hinders the early identification for ovarian cancer and its treatment. To develop a simple and rapid method for quantitative analysis of ovarian cancer, we developed a DNA strand displacement-based method and finished the rapid detection of miR-21 in ovarian cancer cells within 5 min by a one-step isothermal reaction. The fluorescence intensity trajectory had a good linear relationship with miR-21 concentrations in the range of 100 fM-100 nM, with a lower limit of 6.05 pM. This detection method is simple, faster, and accurate. Besides, it can be applied to detect the miRNA biomarkers of other cancers by changing the preset sequences of toehold.

In Vivo Reversal of P-Glycoprotein-Mediated Drug Resistance in a Breast Cancer Xenograft and in Leukemia Models Using a Novel, Potent, and Nontoxic Epicatechin EC31.

The modulation of P-glycoprotein (P-gp, ABCB1) can reverse multidrug resistance (MDR) and potentiate the efficacy of anticancer drugs. Tea polyphenols, such as epigallocatechin gallate (EGCG), have low P-gp-modulating activity, with an EC50 over 10 µM. In this study, we optimized a series of tea polyphenol derivatives and demonstrated that epicatechin EC31 was a potent and nontoxic P-gp inhibitor. Its EC50 for reversing paclitaxel, doxorubicin, and vincristine resistance in three P-gp-overexpressing cell lines ranged from 37 to 249 nM. Mechanistic studies revealed that EC31 restored intracellular drug accumulation by inhibiting P-gp-mediated drug efflux. It did not downregulate the plasma membrane P-gp level nor inhibit P-gp ATPase. It was not a transport substrate of P-gp. A pharmacokinetic study revealed that the intraperitoneal administration of 30 mg/kg of EC31 could achieve a plasma concentration above its in vitro EC50 (94 nM) for more than 18 h. It did not affect the pharmacokinetic profile of coadministered paclitaxel. In the xenograft model of the P-gp-overexpressing LCC6MDR cell line, EC31 reversed P-gp-mediated paclitaxel resistance and inhibited tumor growth by 27.4 to 36.1% (p < 0.001). Moreover, it also increased the intratumor paclitaxel level in the LCC6MDR xenograft by 6 fold (p < 0.001). In both murine leukemia P388ADR and human leukemia K562/P-gp mice models, the cotreatment of EC31 and doxorubicin significantly prolonged the survival of the mice (p < 0.001 and p < 0.01) as compared to the doxorubicin alone group, respectively. Our results suggested that EC31 was a promising candidate for further investigation on combination therapy for treating P-gp-overexpressing cancers.

[Analysis of genome copy number variations in fetuses with isolated ventricular septal defect and a literature review].

OBJECTIVE: To assess the value of copy number variation sequencing (CNV-seq) for revealing the genetic etiology of fetuses with isolated ventricular septal defect (VSD). METHODS: From December 2017 to December 2020, 69 fetuses with isolated VSD were identified at the First Affiliated Hospital of Zhengzhou University. Meanwhile, 839 similar prenatal cases were selected from public databases including Wanfang data, Wanfang Medicine, and China National Knowledge Infrastructure (CNKI) by using keywords such as "Ventricular septal defect", "Copy number variation", and "Prenatal". A total of 908 fetuses with isolated VSD were analyzed. CNV-seq was carried out for 69 fetuses. RESULTS: Among the 908 fetuses, 33 (3.63%) were found to harbor pathogenic CNVs, which included 11 chromosomal aneuploidies (1.21%) and 22 pathogenic CNVs (2.42%). The pathogenic CNVs have involved 12 genetic syndromes, with those known to involve the heart development including 5 cases of 22q11.21 deletion syndrome, 2 cases of 4q terminal deletion syndrome, and 1 case of 9q subtelomere deletion syndrome. The outcome of pregnancies for 15 fetuses with pathogenic CNVs was known, of which 12 were terminated, and 3 had spontaneous closure of the ventricular septum after birth, but 1 of them had other abnormalities. CONCLUSION: Fetuses with isolated VSD have a relatively high risk for chromosomal abnormalities, for which CNV-seq should be recommended.

Whole-exome sequencing in deceased fetuses with ultrasound anomalies: a retrospective analysis.

BACKGROUND: Whole-exome sequencing (WES) is an effective method in the prenatal setting for identification of the underlying genetic etiology of fetal ultrasound abnormalities. To investigate the diagnostic value of WES in fetuses with ultrasound abnormalities that resulted in fetal demise or pregnancy termination. METHODS: 61 deceased fetuses with ultrasound abnormalities and normal copy number variation Sequencing were retrospectively collected. Proband-only or trio-WES were performed on the products of conception. RESULT: Collectively, 28 cases were positive with 39 variants (10 pathogenic, 22 likely pathogenic and 7 variants of uncertain significance) of 18 genes, and the overall diagnostic rate was 45.9% (28/61), of which 39.2% (11/28) were de novo variants. In addition, 21 variants in 11 genes among the positive cases had not been previously reported. The diagnostic yield for definitive findings for trio analysis was 55.9% (19/34) compared to 33.3% (9/27) for singletons. The most common ultrasound abnormalities were skeletal system abnormalities 39.2% (11/28), followed by multiple system abnormalities (17.9%, 5/28) and genitourinary abnormalities (17.9%, 5/28). CONCLUSION: Our results support the use of WES to identify genetic etiologies of ultrasound abnormalities and improve understanding of pathogenic variants. The identification of disease-related variants provided information for subsequent genetic counseling of recurrence risk and management of subsequent pregnancies.

[Analysis of a child with autosomal dominant mental retardation type 40 due to variant of CHAMP1 gene].

OBJECTIVE: To explore the clinical and genetic features of a child with autosomal dominant mental retardation type 40 (MRD40) due to variant of the CHAMP1 gene. METHODS: Clinical characteristics of the child were analyzed. Genetic testing was carried out by low-depth high-throughput and whole genome copy number variant sequencing (CNV-seq) and whole exome sequencing (WES). A literature review was also carried out for the clinical phenotype and genetic characteristics of patients with MRD40 due to CHAMP1 gene variants. RESULTS: The child, a 11-month-old girl, has presented with intellectual and motor developmental delay. CNV-seq revealed no definite pathogenic variants. WES has detected the presence of a heterozygous c.1908C>G (p.Y636*) variant in the CHAMP1 gene, which was carried by neither parent and predicted to be pathogenic. Literature review has identified 33 additional children from 12 previous reports. All children had presented with developmental delay and mental retardation, and most had dystonia (94.1%), delayed speech and/or walking (85.2%, 82.4%) and ocular abnormalities (79.4%). In total 26 variants of the CHAMP1 gene were detected, with all nonsense variants being of loss-of-function type, located in exon 3, and de novo in origin. CONCLUSION: The heterozygous c.1908C>G (p.Y636*) variant of the CHAMP1 gene probably underlay the WRD40 in this child. Genetic testing should be considered for children featuring global developmental delay, mental retardation, hypertonia and facial dysmorphism.

[CNV-seq analysis of copy number variations in 217 fetuses with nasal bone dysplasia].

OBJECTIVE: To assess the diagnostic value of copy number variation sequencing (CNV-seq) in the genetic etiology of fetuses with nasal bone dysplasia (NBD). METHODS: A total of 217 fetuses discovered with NBD from December 2017 to December 2020 were divided into the isolated NBD group and NBD combined with other anomalies group, for which copy number variations (CNVs) were analyzed. RESULTS: A total of 40 fetal abnormalities were detected in 217 cases, with an overall abnormal rate of 18.4%. These included 31 cases with aneuploidies (14.3%, 31/217) and 9 cases with genomic CNVs (4.1%, 9/217). Five cases of trisomy 21 (3.5%, 5/144) and two CNVs cases with unknown clinical significance (1.4%, 2/144) were detected in the isolated group. As for the combined NBD group, 26 aneuploidies (35.6%, 26/73), including 19 cases with trisomy 21, 6 cases with trisomy 18, 1 case with trisomy 13, 5 cases with pathogenic CNVs (6.8%, 5/73), and 2 cases with CNVs of unknown clinical significance (2.7%, 2/73) were detected. A significant difference was detected between the two groups (P < 0.01). CONCLUSION: The detection rate of CNV-seq is high for chromosomal aneuploidies and pathogenic CNVs in fetuses with NBD, particularly in those combined with other ultrasonic abnormalities.

Anticancer activity of synthesized 5-benzyl juglone on selected human cancer cell lines.

BACKGROUND: Cancer is a malignant disease that causes millions of deaths each year worldwide. As one of the cancer therapeutic strategies, chemotherapy is a means to destroy rapidly dividing cells. The main problem with cancer chemotherapy is the lack of selectivity of conventional chemotherapeutic drugs, leading to toxicity towards normal cells. Therefore, the discovery of anticancer agents with selectivity for fast-growing cancer cells was desirable. OBJECTIVE: In this study, we report the synthesis and identification of the novel 5-benzyl juglone as a potential anticancer agent with selectivity toward certain cancer cell lines. METHODS: An efficient synthetic method for 5-benzyl juglone has been established. The proliferation of cancer cell lines and a normal cell line treated by the target compound were studied using an MTT assay. In addition, the cell cycle arrest and apoptosis were determined by flow cytometry. RESULTS: Based on the Diels-Alder (D-A) reaction between 3,6-dimethoxy benzyne intermediate with furan, further acid-catalyzed intramolecular rearrangement and CAN-mediated oxidation, a convenient synthesis of 5-benzyl juglone has been achieved with high overall yield. The results from in vitro biological evaluation indicated that the juglone derivative exhibited potent antiproliferative activity against HCT-15 human colorectal cancer cells with an IC50 value of 12.27 µM. It exerted high inhibitory activity toward MCF-7 human breast cancer cells and, to a much lesser extent, to corresponding MCF-10A human breast epithelial normal cells with the IC50 ratio (IC50 in MCF-7 divided by IC50 in MCF-10A) of 0.62. CONCLUSION: The mechanistic investigations indicated that 5-benzyl juglone could induce cell cycle arrest at the G0/G1 phase and promote apoptosis of HCT-15 cells. The apoptotic effects possibly also contributed to its higher selectivity toward cancer cells than normal cell lines.

[Genetic analysis and prenatal diagnosis for a Chinese pedigree affected with Canavan disease].

OBJECTIVE: To explore the genetic basis for a Chinese patient suspected for Canavan disease. METHODS: Whole exome sequencing (WES) was carried out for the proband, and candidate variants were verified by Sanger sequencing of the proband, her parents and brother. Prenatal diagnosis was provided to her mother by chorionic villi sampling (CVS) upon her subsequent pregnancy. RESULTS: The proband, a 4-month-old female infant, had manifested drowsiness, hypotonia and apathy. Urine metabolism screening showed elevated N-acetylaspartic acid. Cranial magnetic resonance imaging revealed abnormal myelination and multiple abnormal signals in large brain areas. WES revealed that the proband has harbored compound heterozygous variants of the ASPA gene, namely c.187A>G (p.Arg63Gly) in exon 1 and c.634+1G>A (P.?) in exon 4. Sanger sequencing confirmed that the c.187A>G (p.Arg63Gly) and c.634+1G>A (p.?) variants were respectively inherited from her mother and father. Her phenotypically normal brother has carried a heterozygous c.634+1G>A (p.?) variant. Prenatal diagnosis by CVS indicated that the fetus was a heterozygous carrier of the c.187A>G variant. CONCLUSION: WES can facilitate the diagnosis of Canavan disease, particularly for those lacking specific phenotypes of the disease. The compound heterozygous variants of the ASPA gene probably underlay the Canavan disease in this patient, and the result has enabled prenatal diagnosis for this family.

H2A.B is a cancer/testis factor involved in the activation of ribosome biogenesis in Hodgkin lymphoma.

Testis-specific regulators of chromatin function are commonly ectopically expressed in human cancers, but their roles are poorly understood. Examination of 81 primary Hodgkin lymphoma (HL) samples showed that the ectopic expression of the eutherian testis-specific histone variant H2A.B is an inherent feature of HL. In experiments using two HL cell lines derived from different subtypes of HL, H2A.B knockdown inhibited cell proliferation. H2A.B was enriched in both nucleoli of these HL cell lines and primary HL samples. We found that H2A.B enhanced ribosomal DNA (rDNA) transcription, was enriched at the rDNA promoter and transcribed regions, and interacted with RNA Pol I. Depletion of H2A.B caused the loss of RNA Pol I from rDNA chromatin. Remarkably, H2A.B was also required for high levels of ribosomal protein gene expression being located at the transcriptional start site and within the gene body. H2A.B knockdown reduced gene body chromatin accessibility of active RNA Pol II genes concurrent with a decrease in transcription. Taken together, our data show that in HL H2A.B has acquired a new function, the ability to increase ribosome biogenesis.

[Analysis of a case with heterozygous 14q12 deletion and FOXG1 gene-related disease].

OBJECTIVE: To describe the clinical and genetic characteristics of a child with 14q12q13.1 deletion involving the FOXG1 gene. METHODS: Clinical manifestation of the child was analyzed. Peripheral blood sample of the patient was subjected to chromosomal karyotyping and single nucleotide polymorphism array (SNP-array) analysis. RESULTS: The male infant has developed feeding difficulty, poor sucking, lower limb tremor, and frontal bruising 8 days after birth. Magnetic resonance imaging revealed significant enlargement of bilateral ventricles and corpus callosum dysplasia. Chromosomal analysis revealed a karyotype of 46,XY,del(14)(q12q13.1), and SNP-array confirmed that there was a 9.6 Mb deletion in 14q11.2q13.1, which encompassed the FOXG1 gene. CONCLUSION: For patients with brain development abnormalities, dyskinesia, cognitive impairment, speech disorder and other manifestations, copy number variation of the FOXG1 gene should be excluded. SNP-array should be carried out as early as possible to attain the diagnosis.

Chitin nanofibers improve the stability and functional performance of Pickering emulsions formed from colloidal zein.

There is growing interest in formulating Pickering emulsions from biopolymer particles due to consumer demand for more natural products. Protein-based colloidal particles can be used for this purpose, but they are prone to aggregate at pH values around their isoelectric point (pI), which limits their application. In this study, the possibility of using chitin nanofibers (ChNFs) to improve the pH stability of Pickering emulsions prepared from zein colloidal particles (ZCPs) was investigated. Initially, the morphology and interfacial properties of the complexes formed between ChNFs and ZCPs were studied as a function of pH (3-9). The tendency of the ZCPs to aggregate and sediment at pH ≥ pI was reduced in the presence of ChNFs, which was attributed to the formation of electrostatic complexes. The contact angle of the composite particles could be optimized by altering their composition. For instance, the contact angle increased from 74° for ZCPs to 85° for ZCP/ChNF (5:1 ratio) at pH 6, which improved their tendency to stabilize the oil droplets. Brewster angle microscopy indicated that ZCP/ChNF complexes had rod-like and/or particulate structures at an air-water interface, which were different from those observed in the bulk aqueous phase. Pickering emulsions formed from ZCP/ChNF complexes had better stability than those formed from ZCPs or ChNFs, especially when the pH was close to or greater than the pI. An in vitro digestion study showed that the presence of the interfacial complexes reduced the lipolysis of the oil droplets by about 11% in a simulated gastrointestinal tract. High internal phase Pickering emulsions (HIPPEs) could be formed from ZCP/ChNF complexes at pH ≥ pI, which were able to protect unsaturated lipids from oxidation. Overall, our results show that chitin nanofibers can be used to improve the pH stability of Pickering emulsions formed from colloidal zein, as well as to modulate their functional performance.

Epidemiological Investigation and Prevention Control Analysis of the Longitudinal Distribution of COVID-19 in Henan Province, China.

The objective was to analyze the longitudinal distribution, epidemiological characteristics, and local prevention and control measures of coronavirus disease 2019 (COVID-19) in six cities in Henan Province, China, from 21 January 2020 to 17 June 2020: Xinyang City (including Gushi County), Nanyang City (including Dengzhou City), Zhumadian City (including Xincai County), Zhengzhou City (including Gongyi City), Puyang City, and Anyang City (including Hua County). Data were collected and analyzed through the COVID-19 information published on the official websites of the health commissions in the six selected cities of Henan Province. As of 17 June 2020, the cumulative incidence rate of COVID-19 in Henan Province was 1.33/100,000, the cumulative cure rate was 98.27%, the cumulative mortality rate was 1.73%, the age range of diagnosed cases was 5 days to 85 years old, and the male-to-female ratio was 1.09:1. The confirmed cases of COVID-19 in Henan Province were mainly imported cases from Hubei, accounting for 87.74% of all cases, of which the highest proportion was 70.50% in Zhumadian. The contact cases and local cases increased in a fluctuating manner over time. In this paper, epidemiological characteristics of COVID-19 in Henan Province were analyzed from the onset of the outbreak to the effective control within 60 days, and effective and distinctive prevention and control measures in various cities were summarized to provide a favorable useful reference for the further formulation and implementation of epidemic prevention and control and a valuable theoretical basis for effectively avoiding a second outbreak.IMPORTANCE Epidemic prevention and control in China have entered a new stage of normalization. This article analyzes the epidemiological characteristics of COVID-19 in Henan Province and summarizes the effective disease prevention and control means and measures at the prefecture level; the normalized private data provide a theoretical reference for the formulation and conduct of future prevention and control work. At the same time, these epidemic prevention and control findings can also be used for reference in other countries and regions.

[Progress of research on induced pluripotent stem cell models for Down syndrome].

With an incidence of 1/800 - 1/600, Down syndrome (DS) is the most common chromosomal disorder in humans. Whilst most DS patients has trisomy 21, a small proportion may carry translocations or mosaicisms involving chromosome 21. The main characteristics of DS include mental retardation, peculiar facies, growth retardation, congenital heart disease, duodenal stenosis, Alzheimer's disease, leukemia, and immunodeficiency, which may be due to increased dosage of critical genes. Recent studies also showed that epigenetic changes may also occur in DS. For research on patients with DS or other trisomies have been restricted by ethical considerations, and commonly used mouse models cannot fully replicate the characteristic features of DS, pluripotent stem cells induced from fetal samples or biopsy tissues from DS patients may generate models with the same genetic content, which may provide idea materials for studying the pathogenesis of DS and customized cell and/or gene therapies.

DMAKO-20 as a New Multitarget Anticancer Prodrug Activated by the Tumor Specific CYP1B1 Enzyme.

To reduce the pervasive toxicity of natural shikonin, alkannin, and their synthetic analogues and to enhance the selectivity of these chemotherapeutics toward cancer cells, a novel 5,8-dimethyl alkannin oxime derivative (DMAKO-20) was designed, synthesized, and evaluated for its strong antitumor activity both in vitro and in vivo. It showed potent growth inhibitory effects against HCT-15, HCT-116, and K562 cells (IC50 < 1 µM), moderate antiproliferative activity toward MDA-MB-231, HepG2, PANC, Bel7402, and MGC803 cancer cells (IC50 < 10 µM), and was nontoxic to the human normal VEC and HSF cells. In vivo efficacy studies demonstrated that DMAKO-20 (10 mg/kg, i.v. on every the other day, 8 times in 14 days) resulted in 59.3% reduction in HCT-15 xenograft volume. It was as effective as the toxic antimetabolite 5-FU but revealed neither toxicity nor death in mice. The mechanistic investigations indicated that DMAKO-20 underwent the tumor-specific CYP1B1-catalyzed bioactivation to afford nitric oxide and active naphthoquinone mono-oximes, which exhibited combined anticancer effects. It was defined as a representative of the "Multi-target Anticancer Prodrugs Activated by Specific Enzymes in cancer cells". The produced active metabolites exerted anticancer effects by the direct nucleophilic alkylation and the induction of the apoptosis of cancer cells through activation of the mitochondrial pathway. The discovery of DMAKO-20 and the illustration of its molecular mechanisms may provide a new strategy to overcome the nonselective toxicity of the current chemotherapeutics.

Tailoring of structured hydroxypropyl methylcellulose-stabilized emulsions for encapsulation of nobiletin: modification of the oil and aqueous phases.

Hydroxypropyl methylcellulose (HPMC)-stabilized emulsions were produced and their structure was further modified by altering the oil (glycerol monolaurate, GML) and aqueous (whey protein concentrate, WPC) phases. Then, the encapsulation and release of nobiletin were evaluated. HPMC (3%) could efficiently stabilize the oil droplets with a particle size around 370 nm. When HPMC was the emulsifier, the stabilization time of nobiletin in the emulsion was prolonged, but its encapsulation amount was still maintained around 4.5 mg g-1. Nobiletin crystals were found to adsorb on the interface of oil droplets. The addition of GML and WPC changed the microstructure of HPMC-stabilized emulsions. Both of them enhanced the encapsulation efficiency and storage stability of nobiletin. Nobiletin crystals became softer and shorter. After modification, nobiletin was stable for one week at a level of 7.5 mg g-1. The bioaccessibility of nobiletin was modulated in the presence of GML and WPC. The results demonstrate that the structure of emulsion-based delivery systems affects the encapsulation and delivery of hydrophobic components.

Early serodiagnosis of trichinellosis by ELISA using excretory-secretory antigens of Trichinella spiralis adult worms.

BACKGROUND: The excretory-secretory (ES) antigens of Trichinella spiralis muscle larvae (ML) are the most commonly used diagnostic antigens for trichinellosis. Their main disadvantage for the detection of anti-Trichinella IgG is false-negative results during the early stage of infection. Additionally, there is an obvious window between clinical symptoms and positive serology. METHODS: ELISA with adult worm (AW) ES antigens was used to detect anti-Trichinella IgG in the sera of experimentally infected mice and patients with trichinellosis. The sensitivity and specificity were compared with ELISAs with AW crude antigens and ML ES antigens. RESULTS: In mice infected with 100 ML, anti-Trichinella IgG were first detected by ELISA with the AW ES antigens, crude antigens and ML ES antigens 8, 12 and 12 days post-infection (dpi), respectively. In mice infected with 500 ML, specific antibodies were first detected by ELISA with the three antigen preparations at 10, 8 and 10 dpi, respectively. The sensitivity of the ELISA with the three antigen preparations for the detection of sera from patients with trichinellosis at 35 dpi was 100%. However, when the patients' sera were collected at 19 dpi, the sensitivities of the ELISAs with the three antigen preparations were 100% (20/20), 100% (20/20) and 75% (15/20), respectively (P < 0.05). The specificities of the ELISAs with the three antigen preparations were 98.11, 95.60 and 89.31%, respectively (P < 0.05). CONCLUSIONS: The sensitivity and specificity of the T. spiralis AW ES antigens were superior to those of the AW crude antigens and ML ES antigens. Thus, the AW ES antigens might serve as potential antigens for the early and specific serodiagnosis of trichinellosis.