Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA.

Loop-mediated isothermal amplification (LAMP) is a promising diagnostic tool for genetic amplification, which is known for its rapid process, simple operation, high amplification efficiency, and excellent sensitivity. However, most of the existing heating methods are external for completion of molecular amplification with possibility of contamination of specimens. The present research provided an internal heating method for LAMP using magnetic nanoparticles (MNPs), which is called nano-LAMP. Near-infrared light with an excitation wavelength of 808 nm was employed as the heating source; hydroxy naphthol blue (HNB) was used as an indicator to conduct methodological research. We demonstrate that the best temperature was controlled at a working power of 2 W and 4.8 µg/µL concentration of nanoparticles. The lowest limit for the detection of HPV by the nano-LAMP method is 102 copies/mL, which was confirmed by a gel electrophoresis assay. In the feasibility investigation of validated clinical samples, all 10 positive HPV-6 specimens amplified by nano-LAMP were consistent with conventional LAMP methods. Therefore, the nano-LAMP detection method using internal heating of MNPs may bring a new vision to the exploration of thermostatic detection in the future.

Conjugation of ferulic acid onto pectin affected the physicochemical, functional and antioxidant properties.

BACKGROUND: Pectin, as a functional ingredient, is best known as a gelling and thickening agent, stabilizer, and fat substitute in processed foods. The synthesis of pectin graft copolymers is considered one of the most fascinating ways to improve its physicochemical and functional properties. In this study, therefore, water-soluble pectin or ultrasound-treated pectins (UP30 and UP60) grafted with ferulic acid (FA) conjugates, pectin-g-FA, UP30-g-FA and UP60-g-FA, were synthesized via a free radical-mediated grafting procedure, and their physicochemical, structural, and functional characteristics were investigated. Moreover, 2,2-diphenyl-1-picryl-hydrazyl (DPPH)-radical scavenging activity, Trolox equivalent antioxidant capacity (TEAC) assay and ferric-reducing ability of plasma (FRAP) assay were used to evaluate their antioxidant activities in vitro. RESULTS: Results showed that FA was covalently grafted onto pectin or ultrasound-treated pectins, and the grafting ratios of pectin-g-FA, UP30-g-FA and UP60-g-FA were 65.43 ± 1.30, 82.55 ± 1.71 and 75.82 ± 0.89 mg FA/g, respectively. Although the molecular weights, apparent viscosities, and thermal stabilities of the three FA-grafted pectin conjugates decreased and their surface morphologies were different from those of native pectin and ultrasound-treated pectins, they possessed prominent DPPH-radical scavenging ability [half maximal inhibitory concentration (IC50 ) of 0.32 to 0.89 mg mL-1 ) and antioxidant capacity (TEAC of 100.02 to 153.42 µmol Trolox/g sample; FRAP: 166.41 to 270.27 µmol FeSO4 /g sample). Their antioxidant potentials were positively correlated with the grafting ratio. CONCLUSION: This study provided a promising strategy for the functionalization of pectin, and the prepared FA-grafted pectin conjugates could be explored as functional ingredients that showed potential for applications in food and agriculture systems. © 2020 Society of Chemical Industry.

Association between the CYP1A2 polymorphisms and risk of cancer: a meta-analysis.

The previously published data on the association between CYP1A2*1C (rs2069514) and CYP1A2*1F (rs762551) polymorphisms and cancer risk have remained controversial. Hence, we performed a meta-analysis to investigate the association between CYP1A2*1F and CYP1A2*1C polymorphisms and cancer risk under different inheritance models. Overall, significant association was observed between CYP1A2*1F and cancer risk when all the eligible studies were pooled into the meta-analysis (dominant model: OR 1.08, 95 % CI 1.02-1.15; heterozygous model: OR 1.06, 95 % CI 1.01-1.12; additive model: OR 1.07, 95 % CI 1.02-1.13). In the further stratified and sensitivity analyses, for CYP1A2*1F polymorphism, significantly increased lung cancer risk and significantly decreased bladder cancer risk were observed in Caucasians. For CYP1A2*1C polymorphism, no significant association was found in overall and all subgroup analyses. In summary, this meta-analysis suggests that CYP1A2*1F polymorphism is associated with lung cancer and bladder cancer risk in Caucasians.

Preoperative high-intensity strength training combined with balance training can improve early outcomes after total knee arthroplasty.

BACKGROUND: To investigate the effect of preoperative high-intensity strength training combined with balance training on the knee function of end-stage knee osteoarthritis (KOA) patients after total knee arthroplasty (TKA). METHODS: A prospective study was conducted on end-stage KOA patients awaiting TKA. The patients were divided into an experimental group and a control group according to whether they received a preoperative training intervention. The differences in knee flexor-extensor strength, knee range of motion (ROM), timed up and go (TUG) test result, stair ascend/descend test result, Knee Society score (KSS) and Berg balance scale (BBS) score were assessed in both groups at baseline (T1), before operation (T2), 3 months after operation (T3), and 1 year after operation (T4). RESULTS: After high-intensity strength training and balance training, the knee flexor-extensor strength, TUG test result, stair ascend/descend test result, and KSS were all significantly improved at T2 in the experimental group over the control group. At T3, the knee ROM, knee flexor-extensor strength, TUG test result, BBS score, and KSS clinical and functional scores were all significantly superior in the experimental group. The experimental group enjoyed a superiority in KSS clinical and functional scores until T4. Group × time and between-group interactions were found in all assessment indicators in both groups (p < 0.01). CONCLUSION: Preoperative high-intensity strength training combined with balance training can enhance the knee flexor-extensor strength and balance of patients with end-stage KOA in the short term and help improve early outcomes after KOA. Trial registration ChiCTR2000032857, 2020-05-13.

Complete mitochondrial genome of Takydromus kuehnei (Squamata: Takydromus) and its phylogenetic analysis.

We sequenced and annotated the complete mitochondrial genome (mitogenome) of Takydromus kuehnei Van Denburgh, 1909 (Squamata: Takydromus). This mitogenome was 17,224 bp long and encoded 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, one non-coding regions of an L-strand replication origin and a displacement loop region. The overall nucleotide composition was 32.8% of A, 13.8% of G, 24.8% of T, and 30.5% of C. Phylogenetic analysis using maximum likelihood method validated the taxonomic status of T. kuehnei, exhibiting the close relationship with the species from the genus Takydromus.

Physicochemical characteristics and in vitro biological activities of polysaccharides derived from raw garlic (Allium sativum L.) bulbs via three-phase partitioning combined with gradient ethanol precipitation method.

In the present study, three-phase partitioning (TPP) coupled with gradient ethanol precipitation (GEP) was developed for the first time to extract and isolate polysaccharides (GPSs) from raw garlic (Allium sativum L.) bulbs. Four kinds of fructose polymers, namely, GPS35, GPS50, GPS65, and GPS80, were obtained at the final ethanol precipitation concentrations of 35%, 50%, 65%, and 80% (v/v), respectively, and their physicochemical characteristics and in vitro biological activities were investigated. Results indicated that GPS80 had higher carbohydrate (86.68% ± 0.90%) and uronic acid (12.89% ± 0.09%) contents, lower weight-average molecular weight (8.93 × 103 Da), and looser surface morphology than the other three GPSs. Furthermore, among the four GPSs, GPS80 exhibited the strongest antioxidant capacities, inhibitory effects on α-amylase and α-glycosidase, and nitric oxide stimulatory activity on RAW264.7 macrophage cells in vitro. Therefore, this study provides a simple and feasible technological strategy for producing bioactive polysaccharides from raw Allium vegetables.

Production, physicochemical characteristics, and in vitro biological activities of polysaccharides obtained from fresh bitter gourd (Momordica charantia L.) via room temperature extraction techniques.

In this study, polysaccharides (BPSs) were obtained from fresh bitter gourd (Momordica charantia L.) by room temperature extraction techniques, including three-phase partitioning (TPP) and ultrasonic-assisted extraction (UAE) performed in different solvents. The results showed that the extraction methods had significant influence on the extraction yield, chemical composition, weight-average molecular weight (Mw), monosaccharide composition, preliminary structural characterization and microstructure of the BPSs. The BPS-W sample obtained from the bitter gourd residue via UAE in distilled water had a higher uronic acid content (24.22%) and possessed stronger antioxidant capacities and α-amylase and α-glycosidase inhibitory activities than BPS-C extracted with UAE in citric acid, BPS-A extracted with UAE in 1.25 mol/L NaOH/0.05% NaBH4, and BPS-J extracted from bitter gourd juice by TPP. Moreover, BPS-A, which had the lowest Mws, showed the best bile acid-binding capacity among the four BPSs. This study had great potentials for the preparation of bioactive polysaccharides from fresh vegetables.

Efficacy of clomifene citrate for the treatment of patients with polycystic ovary syndrome: A protocol of systematic review.

BACKGROUND: This study aims to assess the efficacy and safety of clomifene citrate (CC) for the treatment of patients with polycystic ovary syndrome (PCOS). METHODS: In this study, we will comprehensively search MEDLINE, EMBASE, The Cochrane Library, Web of Science, CINAHL, ACMD, PsycINFO, and China National Knowledge Infrastructure for original articles published from their inceptions to the January 1, 2020 without language restrictions. All studies will undergo relevance and a design selecting process. Data from qualified studies will be collected by 2 independent authors. Additionally, we will conduct a risk of bias evaluation using a Cochrane risk of bias tool. We will undertake statistical analysis utilizing RevMan 5.3 software. RESULTS: This study will summarize the up-to-date evidence to investigate the efficacy and safety of CC for the treatment of patients with PCOS. CONCLUSION: The findings of this study will provide helpful evidence of CC for the treatment of patients with PCOS, as well as may help develop treatment guidelines. PROSPERO REGISTRATION NUMBER: PROSPERO CRD42020162818.

A combined solid phase extraction/capillary gel electrophoresis method for the determination of phosphorothioate oligodeoxynucleotides in biological fluids, tissues and feces.

A novel biological sample clean-up procedure has been developed for the determination of phosphorothioate oligodeoxynucleotides (PS-ODNs) and derived metabolites in biological fluids (plasma, urine and bile) and in tissues and feces from mice and rats. This method uses a one-step C18 solid-phase extraction (SPE) for biological matrix removal, and it uses capillary gel electrophoresis (CGE) for analyte detection. The assay is specific, and its linearity is superb (r>0.99) for IV-AS (a 13-mer PS-ODN) and PS19 (a 19-mer PS-ODN) in a variety of biological matrices. For both IV-AS and PS19, the precision, accuracy and absolute recovery values were found to be <20%, +/-20% and 80-120%, respectively. The LODs of IV-AS and PS19 were 0.6 mg/l for plasma, 0.8 mg/l for rat urine and bile, 6 microg/g for rat tissues, and 10 microg/g for rat feces, with a signal-to-noise ratio of 3 (S/N=3). This method has been successfully applied to the analysis and quantitation of PS-ODNs in various biological samples arising from preclinical pharmacokinetic studies.

Effect of different drying methods on the product quality and bioactive polysaccharides of bitter gourd (Momordica charantia L.) slices.

In this study, three drying methods, namely, hot-air drying, freeze drying (FD), and infrared radiation drying (ID), were applied to dry bitter gourd (Momordica charantia L.) slices. Results showed that the drying methods had significant influences on appearance, color, rehydration ratio, and microstructure of dried bitter gourd. FD provided high-quality dried bitter gourd products due to the uniform honeycomb network and less collapsed structure. Three water-soluble bitter gourd polysaccharides (BPS-H, BPS-F, and BPS-I) were obtained from the bitter gourd dried using the three drying methods. The three polysaccharides exhibited similar preliminary structural characteristics with different monosaccharide compositions and molecular weights. BPS-I obtained from ID-dried bitter gourd had higher sugar and uronic acid contents than BPS-H and BPS-F. BPS-I exhibited stronger antioxidant activities and bile acid-binding capacity in vitro than BPS-H and BPS-F. Moreover, BPS-F and BPS-I showed significant α-amylase inhibitory activities in vitro.

Preparation, characterization, rheological and antioxidant properties of ferulic acid-grafted curdlan conjugates.

In this study, water-soluble curdlan products (Cur and Cur-D) were prepared by an alkali-neutralization treatment process, after which ferulic acid (FA)-grafted Cur conjugates (Cur-g-FA and Cur-D-g-FA) were fabricated in the presence and absence of salt by adopting an approach involving free-radicals generated by the ascorbic acid/hydrogen peroxide redox pair under an inert atmosphere. Results showed that FA was successfully grafted onto the C-6 and C-4 positions of the Cur chains through covalent linkages and that the presence of salt exerted minor influences on the grafting ratios and structural characterizations of the products. Cur-g-FA and Cur-D-g-FA showed decreased crystallinity, thermal stability, and rheological properties, as well as a distinct surface morphology, when compared with those of native Cur. However, Cur-g-FA and Cur-D-g-FA also exhibited remarkably enhanced free-radical scavenging ability and antioxidant capacity in vitro. These results indicate that FA-grafted Cur conjugates have great potential application in the field of functional foods.

Quaternized curdlan/pectin polyelectrolyte complexes as biocompatible nanovehicles for curcumin.

In this study, a positively charged quaternized curdlan (Qcurd) was used to fabricate polyelectrolyte complex nanoparticles (PEC NPs) with a negatively charged pectin via electrostatic complexation. Results showed that the Qcurd/pectin PEC NPs prepared with 0.5 mg/mL Qcurd and pectin solutions, 1:2 pectin/Qcurd mass ratio, and pH 4.0 in the absence of NaCl were characterized by a spherical morphology in nanoscale, an average particle size of 68 nm, and good dispersibility in aqueous solutions. Curcumin was encapsulated in the Qcurd/pectin PEC NPs through hydrogen bonding with an encapsulation efficiency of ∼82%, a loading content of 13%, and a pH-dependent controlled release. Curcumin-loaded PEC NPs exhibited a significantly enhanced water solubility, excellent free radical scavenging ability and antioxidant capacity in vitro as compared with those of free curcumin.

[Clinical Application of R-ISS Staging System in 412 Newly Diagnosed Patients with Multiple Myeloma].

OBJECTIVE: To evaluate the prognostic value of R-ISS staging system in patients with newly diagnosed multiple myeloma (NDMM). METHODS: The Chinical data of 412 patients with NDMM in our hospital from May 2010 to May 2016 were retrospectively analyzed. All the patients received conventional chemotherapy or thalidomide or bortezomib-based chemotherapy. All the patients with NDMM were divided into R-ISS-Ⅰ, R-ISS-Ⅱ and R-ISS-Ⅲ groups according to R-ISS staging system on the basis of ISS staging system, cytogenetics and LDH level. The progression-free survival (PFS) time and overall survival(OS) of different groups were compared. RESULTS: Among all 412 patients, 76 were rated as R-ISS-Ⅰ, 259 as R-ISS-Ⅱ and 77 as R-ISS-Ⅲ. The median PFS time in 3 groups were 44, 25 and 14 months respectively (P<0.01). The median OS time of the 3 groups were not reached 54 and 25 months respectively (P<0.01). Further analysis also found that statistically different survival associated with different R-ISS groups in the conventional chemotherapy group (P<0.05), bortezomib-based chemotherapy group (P<0.01), thalidomide-based chemotherapy group (P<0.01), transplantation group (P<0.05), different-age stratified group (≤65y P<0.01, 66-75y P<0.01,≥76y P<0.01), damaged renal function group (P<0.01) and extramedullary infiltration group (P<0.01). CONCLUSION: PFS and OS in the patients with multiple myeloma were different among three distrinct R-ISS stages. The R-ISS staging system has important clinical significance for the prognosis evaluation of multiple myeloma.

Conformational and rheological properties of a quaternary ammonium salt of curdlan.

This study investigated the chain conformation, microstructure, and rheological properties of a quaternary ammonium salt of curdlan (Qcurd) in aqueous medium. Results showed that Qcurd exhibited typical polyelectrolyte behavior in pure water, and the normal viscosity character was observed in 0.1 M NaCl aqueous solution. The weight-average molecular weight (Mw), z-average radius of gyration (Rg), hydrodynamic radius (Rh), conformational parameter (α), characteristic constant (a) and structure parameter (ρ) for Qcurd were determined as 8.08 × 104 g/mol, 26.7 nm, 15.0 nm, 0.54, 0.62, and 1.78, respectively, indicating that Qcurd existed as a flexible chain conformation in 0.1 M NaCl solution, verified by atomic force microscopy. Qcurd exhibited more pronounced shear-thinning behavior at higher concentrations. The flow behavior of Qcurd at different concentrations and temperature was well evaluated by power-law model and Arrhenius equation. Dynamic tests revealed that Qcurd showed oscillatory behaviors between a dilute solution and a weak elastic gel.

Subcritical water extraction-based methods affect the physicochemical and functional properties of soluble dietary fibers from wheat bran.

In this study, subcritical water extraction (SWE), SWE in aqueous citric acid (pH 5.0) (SWEC), and ultrasound-assisted SWEC (USWEC) were used to extract soluble dietary fiber (SDF) from wheat bran. Results showed that SWE-based methods significantly influenced the physicochemical, functional, and biological properties of the SDF. The fraction SDF-III attained via USWEC had higher SDF yield (46.30%) and carbohydrate content (82.91%), and lower weight-average molecular weight (65.2 kDa) and particle size (1.17 µm), and looser and more porous surface structure, compared with the SDF-I and SDF-II obtained by SWE and SWEC, respectively. USWEC increased the thermal stability and homogeneity of SDF-III but decreased its apparent viscosity and dynamic viscoelasticity. Moreover, the SDF-III exhibited more significant antioxidant and α-amylase inhibitory activities in vitro than SDF-I and SDF-II. Therefore, the USWEC technique had a greater potential for the highly-efficient production of SDF from wheat bran.

A Systematic Inflammation-based Model in Advanced Pancreatic Ductal Adenocarcinoma.

Emerging evidence revealed the critical role of systematic inflammation in pancreatic ductal adenocarcinoma (PDAC). In the present study, we reviewed the records of 279 patients with advanced PDAC. Among them, 147 cases were used as the training cohort and another 132 as the validation cohort. In the training cohort, distant metastasis, carbohydrate antigen 19-9 (CA19-9), Glasgow prognostic score (GPS), neutrophil-to-lymphocyte ratio (NLR), and lymphocyte-to-monocyte ratio (LMR) were independent prognostic factors in Cox regression. A nomogram based on these factors was generated to predict median survival time and survival probabilities at 6, 12, and 18 months. The nomogram showed a better discriminatory ability than the American Joint Committee on Cancer (AJCC) TNM staging (C-index: 0.727 vs. 0.610). In the validation cohort, a nomogram composed of the same variables also showed a high discriminatory ability (C-index: 0.784). In the low-risk group with a nomogram total point (NTP) value of more than 175, patients receiving combination therapy showed better prognosis than those receiving monotherapy (P=0.015). In conclusion, the nomogram based on inflammatory biomarkers can serve as useful prognostic tool for advanced PDAC. In addition, patients with high NTP can greater benefit from combination chemotherapy than monotherapy.

Formation and characterization of polyelectrolyte complex synthesized by chitosan and carboxylic curdlan for 5-fluorouracil delivery.

In this study, negatively charged carboxylic curdlan (Cc) bearing a ß-1,3-polyglucuronic acid structure was employed to fabricate nanosized polyelectrolyte complexes (PECs) with positively charged chitosan (CS) in aqueous solution as potential carriers for 5-fluorouracil (5Fu) delivery. Nanosized CS/Cc PECs were formed by the addition of 0.5mg/mL solutions of CS and Cc with a mixing ratio of 1:1 (w/w) at pH 3.0. Under optimized conditions, the prepared CS/Cc PECs showed spherical morphology with an average size of about 180nm and a zeta potential of around 41mV. The 5Fu drug was incorporated into the nanosized CS/Cc PECs and showed excellent encapsulation efficiency (86.47%) and loading content (10.81%). The drug release data in vitro indicated that the nanosized CS/Cc PECs are promising carriers for the sustained release of 5Fu with an anomalous transport mechanism following the Ritger-Peppas model. Besides, the CS/Cc PECs exhibited low cytotoxic activity against SPCA-1 and HeLa cell lines in vitro. This finding suggested that the development of the nanosized CS/Cc PECs offered great promise as an antitumor drug platform.

pH dependent green synthesis of gold nanoparticles by completely C6-carboxylated curdlan under high temperature and various pH conditions.

A C6-carboxylated curdlan (C6-Cc) obtained from 4-acetamido-TEMPO-mediated oxidation of curdlan was used both as a reducing and stabilizing agent for green synthesis of pH-responsive AuNPs, which was carried out by controlling the pH of the C6-Cc solution at a high temperature (100°C). C6-Cc presented a semi-flexible random coil chain in the aqueous medium at pH 5.5 and became more expanded and rigid in alkaline conditions (pH 7.1-12.0), though the primary chemical structure of C6-Cc was virtually unchanged with the pH variation. The AuNPs prepared with C6-Cc at various pHs were characterized by various instrumental measurements. The shapes and sizes of AuNPs were found to be strongly dependent on the pH of the C6-Cc solution. The C6-Cc-decorated AuNPs exhibited a more well-dispersed spherical morphology with smaller particle sizes under alkaline conditions (pH 7.1-12.0). Through this study, a facile, simple, and green method has been demonstrated for preparation of stimuli-sensitive AuNPs using biocompatible polyanionic polysaccharides.

Purification, structural characterization and bioactivity evaluation of a novel proteoglycan produced by Corbicula fluminea.

A novel proteoglycan, named CFPS-11, was isolated from Corbicula fluminea, which is a food source of freshwater bivalve mollusk. CFPS-11 had an average molecular weight of 807.7kDa and consisted of d-glucose and d-glucosamine in a molar ratio of 12.2:1.0. The protein moiety (∼5%) of CFPS-11 was covalently bonded to the polysaccharide chain in O-linkage type through both serine and thereonine residues. The polysaccharide chain of CFPS-11 was composed of (1→4)-α-d-glucopyranosyl and (1→3,6)-α-d-glucopyranosyl residues, which branched at O-6. The branch chain consisted of (1→)-α-d-glucopyranosyl and (1→)-α-d-N-acetylglucosamine residues. CFPS-11 exhibited significant antioxidant activity in a dose-dependent manner and remarkable inhibition activities against α-amylase and α-glucosidase by in vitro assays. These findings indicated that the CFPS-11 from C. fluminea has the potential for development as a health food ingredient.

Quantification of the polyphenols and triterpene acids in chinese hawthorn fruit by high-performance liquid chromatography.

The levels of seven polyphenols (epicatechin, procyanidin B2, procyanidin B5, procyanidin C1, hyperoside, isoquercitrin, and chlorogenic acid) and two triterpene acids (oleanolic acid and ursolic acid) in the matured fruits of Chinese hawthorn (Crataegus pinnatifida Bge. var. major N.E.Br.) were determined by high-performance liquid chromatography methods. The average contents of those constituents in 37 representative cultivars were 1405, 1505, 339, 684, 56, 41, 234, 952, and 147 microg/g fresh weight (FW), respectively. A significant inverse correlation between the procyanidin contents and the latitude of the geographical origin of the cultivars was observed (r = 0.3851, P < 0.02). Correlation analysis of the levels of the nine compounds in the 37 cultivars yielded a strong correlation (P < 0.001) between the individual levels of the four procyanidins and the sum of the procyanidins level (r = 0.7413-0.9898) and between the flavonoids and the chlorogenic acid (r = 0.5383-0.9212). The changes in level of the nine compounds in the hawthorn fruit were evaluated during maturation using the Hebei Dajinxing cultivar. Sixty-one days after blossom, the polyphenol level reached the highest point and the sum of the contents was 1.36 g/100 g FW.