Next-generation sequencing specifies Angiostrongylus eosinophilic meningoencephalitis in infants: Two case reports.

RATIONALE: Angiostrongylus cantonensis-induced eosinophilic meningoencephalitis (AEM) in infants is a very rare but fatal disease. Utilization of genetic assay to detect the cerebral parasite plays an important role for the treatment of the infection. PATIENT CONCERNS: Two infants (<2 years) presented with cough, intermittent fever, mental fatigue, and poor diet. DIAGNOSIS: The patients were under clinical examination and laboratory test including cardiac ultrasound, chest X-ray, blood or cerebrospinal fluid (CSF) cell counting, serum enzyme-linked immunosorbent assay (ELISA), head magnetic resonance imaging (MRI) and next-generation sequencing (NGS) on DNA from CSF. Due to hypereosinophils in patients' peripheral blood and CSF, and abundant DNA sequences from A cantonensis in CSF, the patients were diagnosed with Angiostrongylus eosinophilic meningoencephalitis. INTERVENTIONS: The patients were treated with albendazole to deworm, and methylprednisolone to reduce inflammation. OUTCOME: The patients were completely recovered from AEM without relapse after 10-day treatment. LESSONS: ELISA and MRI are not sufficiently accurate for the diagnosis of AEM in infants. NGS can specify the infection by the cerebral parasite and offers a new effective approach for the early and precise diagnosis of AEM in infants.

Deep eutectic solvent as a green solvent for enhanced extraction of narirutin, naringin, hesperidin and neohesperidin from Aurantii Fructus.

INTRODUCTION: In the present study, a green and efficient extraction method using deep eutectic solvents as extraction solvent was developed for extracting the four major active compounds narirutin, naringin, hesperidin and neohesperidin from Aurantii Fructus. METHODOLOGY: A series of tunable deep eutectic solvents were prepared and investigated by mixing choline chloride or betaine to different hydrogen-bond donors, and betaine/ethanediol was found to be the most suitable extraction solvent. To achieve the best extraction yield, the primary factors affecting the extraction efficiency, such as hydrogen-bond acceptor/hydrogen-bond donor ratio, water content in deep eutectic solvents, extraction temperature, solid/liquid ratio and extraction time, were investigated. RESULTS: The optimal extraction conditions were 40% of water in betaine/ethanediol (1:4) at 60°C for heated extraction of 30 min and solid/liquid ratio 1:100 g/mL. Under the optimum extraction condition, the extraction yields of narirutin, naringin, hesperidin, and neohesperidin were 8.39 ± 0.61, 83.98 ± 1.92, 3.03 ± 0.35 and 35.94 ± 0.63 mg/g, respectively, which were much higher than those of methanol as extraction solvent (5.5 ± 0.48, 64.23 ± 1.51, 2.16 ± 0.15 and 30.14 ± 0.62 mg/g). CONCLUSION: The present results showed that deep eutectic solvents could be promising green and efficient solvents for extraction of the bioactive ingredients from traditional Chinese medicine.

Isolation and identification of antiproliferative compounds from the roots of Tetrastigma hemsleyanum against MDA-MB-435S cell lines.

This present study aimed to elucidate antiproliferative activity of four extracts (CHCl(3), EtOAc, n-BuOH and H(2)O) and chemical constituents isolated from the most potent extract of Tetrastigma hemsleyanum Diels et. Gilg (TDG) against MDA-MB-435S cell lines using the MTT assay at various concentrations in vitro. Ten compounds were isolated and identified as (1) ß-sitosterol, (2) palmitic acid, (3) protocatechuic acid, (4) salicylic acid, (5) p-hydroxybenzoic acid, (6) resveratrol, (7) trans-4-hydroxycinnamic acid, (8) kaempferol, (9) quercetin, and (10) isoquercitrin. Compounds 3, 5-7, 10 were the first report of isolation from this plant. Moreover, antiproliferative activity displayed that the CHCl(3), H(2)O extracts and compounds 6, 8 exhibited obvious inhibitory effects on MDA-MB-435S cell lines with IC(50) values 100.28± 2.64, 127.48±3.45, 92.39±1.68 and 120.30±1.97µ/mL, respectively. Thus the obtained results indicate antiproliferative activity of TDG against MDA-MB-435S cell lines is ascribable to the most potent CHCl(3) extract along with active compounds 6 and 8, which could be considered as a potential chemotherapeutic agent in breast cancer.

Gestational N-hexane inhalation alters the expression of genes related to ovarian hormone production and DNA methylation states in adult female F1 rat offspring.

Research has revealed that n-hexane can disrupt adult female endocrine functions; however, few reports have focused on endocrine changes in adult F1 females after maternal exposure during gestation. In this study, female Wistar rats inhaled 100, 500, 2500, or 12,500 ppm n-hexane for 4 h daily during their initial 20 gestational days. The F1 female offspring exhibited abnormal oestrus cycles. Compared with the controls, the in vitro-cultured ovarian granulosa cells of the 12,500 ppm group showed significantly reduced in vitro progesterone and oestradiol secretion. Elevated progesterone secretion was observed in the 500 ppm group, and decreased and significantly upregulated mRNA expression of the Star, Cyp11a1, Cyp17a1, and Hsd3b genes was observed in the 12,500 ppm and 500 ppm groups, respectively. The protein expression levels were consistent with the mRNA expression levels. Methylation screening of the promoter regions of these genes was performed using MeDIP-chip and confirmed by methylation-sensitive high-resolution melting (MS-HRM), and the observed methylation state changes of the promoter regions were correlated with the gene expression levels. The results suggest that the hormone levels in the female offspring after gestational n-hexane inhalation correspond to the expression levels and DNA methylation states of the hormone production genes.

Chemical constituents from Sedum aizoon and their hemostatic activity.

CONTEXT: Sedum aizoon L. (Crassulaceae) (SA) is widely used to treat various hemorrhages in folk medicine. However, its hemostatic constituents are not yet clear. OBJECTIVE: The chemical constituents of EtOAc fraction from SA and their hemostatic activity were investigated to provide a basis for the application in folk use. MATERIALS AND METHODS: The chemical constituents were isolated from the aerial parts of SA by column chromatography and identified by IR, MS, and NMR, then tested for hemostatic activity using the capillary method and coagulation assays including blood clotting time in vivo, and prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) in vitro at concentrations of 300.0, 100.0, and 30.0 µg/mL. RESULTS: Eleven compounds were identified as p-hydroxybenzoic acid (1), gallic acid (2), protocatechuic acid (3), vallinic acid (4), thymine (5), caffeic acid (6), 5,7-dihydroxy chromone (7), pyrogallol (8), quercetin (9), kaempferol (10), and luteolin (11). This is the first report of compounds 3-8 being isolated from this plant. Compounds 2 (300.0 and 100.0 µg/mL), 4 (100.0 µg/mL), and 11 (100.0 and 30.0 µg/mL) significantly reduced the clotting time (p < 0.01) with inhibition rates of 34.7, 24.5, 30.3, 25.9, and 36.6%, respectively. For further mechanism study, they also reduced PT (3.5, 2.5, 3.5, 3.5, and 3.8%, respectively), APTT (4.5, 3.3, 11.4, 8.5, and 11.1%, respectively), and TT (20.3, 3.8, 7.6, 6.1, and 10.3%, respectively). DISCUSSION AND CONCLUSION: SA produced hemostatic activity possibly related to the presence of gallic acid, vallinic acid, and luteolin, which may be potent candidates of hemostatic drug.

Hylaranins: prototypes of a new class of amphibian antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii.

Amphibian skin secretions contain a broad spectrum of biologically active compounds, particularly antimicrobial peptides, which are considered to constitute a first line of defence against bacterial infection. Here we describe the identification of two prototype peptides representing a novel structural class of antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii. Named hylaranin-L1 (GVLSAFKNALPGIMKIIVamide) and hylaranin-L2 (GVLSVIKNALPGIMRFIAamide), both peptides consist of 18 amino acid residues, are C-terminally amidated and are of unique primary structures. Their primary structures were initially deduced by MS/MS fragmentation sequencing from reverse-phase HPLC fractions of skin secretion that demonstrated antimicrobial activity. Subsequently, their precursor-encoding cDNAs were cloned from a skin secretion-derived cDNA library and their primary structures were confirmed unequivocally. Synthetic replicates of both peptides exhibited broad-spectrum antimicrobial activity with mean inhibitory concentrations (MICs) of 34 µM against Gram-negative Escherichia coli, 4.3 µM against Gram-positive Staphylococcus aureus and 4-9 µM against the yeast, Candida albicans. Both peptides exhibited little haemolytic activity (<6%) at the MICs for S. aureus and C. albicans. Amphibian skin secretions thus continue to provide novel antimicrobial peptide structures that may prove to be lead compounds in the design of new classes of anti-infection therapeutics.

[Chemical constituents from ethyl acetate extract of Sedum aizoon].

OBJECTIVE: To study the chemical constituents from ethyl acetate extract of Sedum aizoon. METHODS: The chemical constituents were isolated by column chromatography and identified by MS and NMR. RESULTS: 11 compounds were isolated and identified as: protocatechuic acid (1), caffeic acid (2), 5,7-dihydroxy chromone (3), methyl gallate (4), ethyl gallate (5), myricetin (6), luteolin (7), cynaroside (8), 3',4',5,7-tetrahydroxy flavanone (9), iriflophene (10) and iriflophene-2-O-ß-D-glucoside (11). CONCLUSION: Compounds 1-3 and 8-11 are the first report of isolation from this plant.

[Protection of apoptosis of osteoblast cultured in vitro by Morinda Root Polysaccharide].

OBJECTIVE: To explore the protection on apoptosis and the mechanism of promoting the cytoactive of osteoblast by Morinda Root Polysaccharide through the observations of the cultured osteoblast in vitro. METHODS: Prepared blood serum with Morinda Root Polysaccharide and Morinda Root aqueous extract and cultured Osteoblast in vitro with it. The second generation osteoblasts in vitro were separated from the cranium of 24-hours newborn SD rat, which were divided into control group (adding only rat serum during cultivation), induction apoptosis group (adding trans-retinoic acid in control group), Morinda Root aqueous extract group (adding serum prepared by Morinda Root aqueous extract in induction apoptosis group) and Morinda Root Polysaccharide group (adding serum prepared by Morinda Root Polysaccharide in induction apoptosis group). Adopting fluorescence microscope, apoptosis detected by flow cytometry and gene expression of Bcl-2 and Bax detected by RT-PCR, to evaluate the effect of Morinda Root Polysaccharide on the course of osteoblast apoptosis. RESULTS: The apoptotic rate of Morinda Root aqueous extract group and Morinda Root Polysaccharide group were significantly lower than that of induction apoptosis group (P < 0.01). The apoptosis ratio of Morinda Root Polysaccharide group was lower than that of Morinda Root aqueous extract group (P < 0.05). Expression level of Bcl-2 mRNA of apoptosis cell: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01). Expression level of Bax mRNA: induction apoptosis group > Morinda Root aqueous extract group > control group > Morinda Root Polysaccharide group (P < 0.01). Bcl-2/Bax: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01). CONCLUSION: Morinda Root can inhibit the apoptosis of osteoblast induced by trans-retinoic acid in some extent. The above role of Morinda Root Polysaccharide is significant better than that of Morinda Root aqueous extract. It is indicated that Morinda Root Polysaccharide is one of the essential component of inhibiting osteoblast apotosis.

[Study on mechanism of yangxincao capsule in regulating lipid metabolism].

OBJECTIVE: To investigate the effect and mechanism of Yangxincao Capsule (YXCC) in regulating lipids. METHODS: Sixty rats were randomly divided into 6 groups, the normal control group (A), the hyperlipidemia model group (B), the high, middle and low dose YXCC treated groups (C, D and E), and the Shanzhajing (SZJ) treated group (F) for positive medicine control. Except for the rats in the normal control group, the other 50 were daily fed with fatty emulsion for 10 days to establish hyperlipidemic model. From the I th day on, in the same time of continually feeding with fatty emulsion they were administered with water, high (1.08 g/kg), middle (0.54 g/kg), low dose (0.27 g/kg) of YXCC and SZJ (5.4 mg/kg) respectively for 10 days, while to rats in Group A equal volume of water was given. At the 21th day, after rats were fasted for 16 h, their blood was extracted from post-orbital vein to detect the level of serum lipids, lipoprotein, apolipoprotein (apo) and lipid metabolic enzyme. RESULTS: Compared with Group A, the levels of serum total cholesterol (TC), triglyceride (TG) and low density lipoprotein-cholesterol (LDL-C) increased remarkably, and the level of high density lipoprotein-cholesterol (HDL-C) dropped obviously in Group B. While in the four treated groups the levels of TC, TG and LDL-C were significantly reduced, HDL-C and its sub-components 2 and 3 (HDL2-C and HDL3-C), as well as the ratio of HDL-C/TC were raised. Besides, the content of apo-Al was increased and apo-B was decreased significantly in Group C and D, activity of lecithin cholesterol acyltransferase (LCAT) and lipoprotein lipase (LPL) increased in the three YXCC treated groups, all showed statistical significance (P < 0.05 or P < 0.01) as compared with those in Group B. CONCLUSION: YXCC could remarkably modulate the lipid metabolic disorder in hyperlipidemic rats, and has a certain bi-directional regulating function on lipoprotein, inferring that it could reduce the risk of occurring coronary artery diseases. The mechanism of regulating lipid metabolism might be related with the increasing activity of LCAT, LPL and eliminating of cholesterol by the elevated level of HDL2-C.