High-sensitivity, ultrawide linear range, antibacterial textile pressure sensor based on chitosan/MXene hierarchical architecture.

It is still a great challenge for the flexible piezoresistive pressure sensors to simultaneously achieve wide linearity and high sensitivity. Herein, we propose a high-performance textile pressure sensor based on chitosan (CTS)/MXene fiber. The hierarchical "point to line" architecture enables the pressure sensor with high sensitivity of 1.16 kPa-1 over an ultrawide linear range of 1.5 MPa. Furthermore, the CTS/MXene pressure sensor possesses a low fatigue over 1000 loading/unloading cycles under 1.5 MPa pressure load, attributed to the strong chemical bonding between CTS fiber and MXene and excellent mechanical stability. Besides, the proposed sensor shows good antibacterial effect benefiting from the strong interaction between polycationic structure of CTS/MXene and the predominantly anionic components of bacteria surface. The sensor is also applied to detect real-time human action, an overall classification accuracy of 98.61% based on deep neural network-convolutional neural network (CNN) for six human actions is realized.

CRISPR/Cas12a-Assisted isothermal amplification for rapid and specific diagnosis of respiratory virus on an microfluidic platform.

Respiratory viruses have long been a major cause of a global pandemic, emphasizing the urgent need for high-sensitivity diagnostic tools. Typical PCR technology can only determine the type of virus in the sample, which is unable to detect different variants of the same virus without costly and time-consuming gene sequencing. Here, we introduce a simple, fully enclosed, and highly integrated microfluidic system based on CRISPR/Cas12a and isothermal amplification techniques (LOC-CRISPR) that can specifically identify multiple common respiratory viruses and their variants. The LOC-CRISPR chip integrates viral nucleic acid extraction, recombinant polymerase amplification, and CRISPR/Cas12a cleavage reaction-based detection, contamination-free detection. In addition, the LOC-CRISPR chip was designed for multiplexed detection (two-sample input and ten-result outputs), which can not only detect the presence of SARS-CoV-2, H1N1, H3N2, IVB and HRSV but also differentiate the BA.1, BA.2, and BA.5 variants of SARS-COV-2. For clinical validation, the LOC-CRISPR chip was used to analyze 50 nasopharyngeal swab samples (44 positive and 6 negative) and achieved excellent sensitivity (97.8%) and specificity (100%). This innovative LOC-CRISPR system has the ability to quickly, sensitively, and accurately detect multiple target nucleic acid sequences with single-base mutations, which will further improve the rapid identification and traceability of respiratory viruses infectious diseases.

Recent advances in the rapid detection of microRNA with lateral flow assays.

MicroRNAs (miRNA) are a kind of endogenous non-coding small molecule RNA, which are widely found in plants, animals, and viruses. miRNA can regulate the post-transcriptional cleavage of target message RNA or inhibit its translation process, and plays an important regulatory role in gene expression. Further studies have shown that miRNAs also play an important role in tumorigenesis and host-pathogen interactions. Given the important contribution of miRNAs in gene regulation and biological functions, rapid and accurate detection of miRNAs has become increasingly significant. Lateral flow assay is a detection method derived to achieve rapid detection in the field. It overcomes the problems of cumbersome, time-consuming, and high cost in traditional detection methods, and provides a good platform for accurate, sensitive, and immediate detection of target analytes. With the advantages convenient, cost-saving, and repaid result readout, the lateral flow test strips are now a recognized point-of-care testing (POCT) device and has been widely used in medical diagnosis, agriculture, food, and environmental safety. For the first time, we summarized the research progress of miRNA detection based on lateral flow methods, focusing on the strategies used in the lateral flow assays for miRNA detection and introducing some main experimental operation details, as well as providing an outlook on the future development direction of this field, aiming to provide a general guidance for colleagues preparing to conduct related research.

Rapid identification of Amanita citrinoannulata poisoning using colorimetric and real-time fluorescence and loop-mediated isothermal amplification (LAMP) based on the nuclear ITS region.

Health concerns and financial losses caused by mushroom poisoning have been reported worldwide. Amanita citrinoannulata, a poisonous mushroom commonly found in China, results in a toxic reaction in humans after mistaken ingestion. To reduce the mistaken ingestion of poisonous mushrooms and to improve clinical diagnosis of mushroom poisoning, a rapid mushroom species identification method is required. Such identification methods could be advantageous in the identification of other poisonous mushroom species. This study developed two rapid and sensitive methods for the detection of A. citrinoannulata utilizing colorimetric and real-time loop-mediated isothermal amplification (LAMP) technology and specifically designed primers for the internal transcribed spacer (ITS) genes of A. citrinoannulata. The methods demonstrated high sensitivity as 0.2 ng of A. citrinoannulata DNA could be detected, with no cross-reaction with 41 non-target mushroom species. The entire detection process could be completed within 40 min without requiring complex instruments and can be observed by the naked eye. Therefore, these novel methods can be used for the identification of fresh and cooked mushroom samples and vomit samples, which contain only 1% A. citrinoannulata. Furthermore, these methods facilitate the detection of mushroom poisoning, and thus, have potential to reduce the number of mushroom poisoning-related deaths worldwide.

Rapid identification of Takifugu genus using visual loop-mediated isothermal amplification.

Some Takifugu species are commonly found in the coastal areas of China, Japan, Thailand, and Korea and cause pufferfish poisoning, which is toxic and even lethal to humans. From 2010 to 2015, there were 430 cases of pufferfish poisoning worldwide, resulting in 52 deaths. Identification of Takifugu species is imperative to reduce financial losses and ensure food safety. Here, visual loop-mediated isothermal amplification (LAMP) was applied to identify Takifugu species. Conserved regions within the mitochondrial DNA among different Takifugu species were selected to design LAMP primers. In 55 min of amplification, sufficient DNA was obtained to observe the results with the naked eye, without the need for complicated instruments. The method was highly specific, with no cross-detection of 17 other fish species, namely, 7 Tetraodontiformes species and 10 commercially important fish. The method showed a detection limit of 0.1 ng Takifugu DNA and was successfully validated to detect Takifugu in cooked fish and the vomitus of poisoned patients. This rapid and visual LAMP method is a useful tool to prevent false labeling, protect consumer rights, and reduce the risk of pufferfish poisoning. PRACTICAL APPLICATION: The loop-mediated isothermal amplification method established in this study can identify cooked or digested fish products containing 1% or more of Takifugu. Therefore, it can be used for the visual detection of Takifugu products and the medical diagnosis of Takifugu poisoning.

Rapid Russula senecis identification assays using loop-mediated isothermal amplification based on real-time fluorescence and visualization.

Russula senecis, a common poisonous mushroom, is widely distributed in China. Mushroom poisoning is becoming a major threat to human health and its rate is increasing worldwide. For the first time, we developed a set of loop-mediated isothermal amplification (LAMP) assays based on a real-time fluorescence and a visualization method to detect R. senecis, and the visual LAMP reaction system was optimized to further shorten the reaction time. Both real-time LAMP and visual LAMP could detect as low as 3.2 pg of genomic DNA. In addition, fried and digested mushrooms were used to validate the proposed LAMP method, and mushroom mixtures with as low as 1% of the target species could be successfully detected, indicating that the LAMP assays established in this study had good applicability and could be used for clinical sample detection and forensic identification. Furthermore, the LAMP assays were proven to be comparable to the real-time PCR method. KEY POINTS: • A set of loop-mediated isothermal amplification (LAMP) assays based on real-time fluorescence and visualization to detect Russula senecis was developed. • Both real-time LAMP and visual LAMP can be used to detect genomic DNA at concentrations as low as 3.2 pg. • By simulating mushroom processing and digestion in gastric juice, LAMP assays were proved to have good applicability and could be used for clinical diagnosis and forensic analysis.

A strip of lateral flow gene assay using gold nanoparticles for point-of-care diagnosis of African swine fever virus in limited environment.

Recombinase polymerase amplification (RPA) was combined with lateral flow to develop a gold nanoparticles test strip for point-of-care diagnosis of African swine fever virus (ASFV), which is called lateral flow gene assay (LFGA). Common diagnostic techniques, including polymerase chain reaction (PCR) and immunochromatography, are time-consuming and labor-intensive, and generally require costly instruments. For improvement, this assay used tailed primers to produce DNA duplexes with a single-stranded tail at one end which can hybridize with a gold nanoparticle (AuNP)-labeled oligonucleotide detection probe. And then, biotin attached to the other end of the product bound to streptavidin, which previously fixed to the test line. Therefore, there would form a sandwich structure, and gold nanoparticles labeled on the detection probe would show a red band on the test line of strip. With the low reaction temperature (37~42 °C) and short reaction time (30 min), LFGA can specifically identify ASFV in blood samples infected with ASFV and classical swine fever virus (CSFV), and the LOD was 102 copies/µL, which was comparable to that of agarose gel electrophoresis. In addition, blood samples infected with ASFV and CSFV were tested, and it was found that the LFGA can specifically identify ASFV DNA. In conclusion, LFGA achieves visual observation of the product after rapid RPA amplification and does not require any expensive instruments during the entire process, which is very helpful for early diagnosis of ASFV. Combined recombinase polymerase amplification (RPA) with lateral flow, we developed a gold nanoparticles test strip for point-of-care diagnosis of African swine fever virus. The upstream primers of RPA were modified with biotin, and the downstream primers were modified with a C3 spacer and an oligonucleotide tail that can be hybridized to a gold nanoparticle-labeled oligonucleotide detection probe. On the strip, the test line and control line were sprayed with streptavidin and an oligonucleotide control probe. In the presence of positive products, RPA products can form a sandwich structure on the test line. Therefore, two red lines will be displayed both on the test line and control line. When there is no positive product, only the control line is shown in red. Its low reaction temperature (37~42 °C) and short time of amplification and detection (30 min) make ASFV realizing point-of-care diagnosis in limited environment.

Rapid and Visual Identification of Chlorophyllum molybdites With Loop-Mediated Isothermal Amplification Method.

Chlorophyllum molybdites is a kind of common poisonous mushroom in China that is widely distributed in different areas. Food poisoning caused by accidentally eating C. molybdites has become more frequent in recent years. In 2019, there were 55 food poisoning incidents caused by eating this mushroom in China. Mushroom poisoning continues to be a common health issue of global concern. When mushroom poisoning occurs, an effective, simple, and rapid detection method is required for accurate clinical treatment or forensic analysis. For the first time, we established a loop-mediated isothermal amplification (LAMP) assay for the visual detection of C. molybdites. A set of specific LAMP primers was designed, and the specificity was confirmed against 43 different mushroom species. The LAMP method could detect as low as 1 pg of genomic DNA. Boiled mushrooms and artificial gastric-digested mushroom samples were prepared to test the applicability of the method, and the results showed that as low as 1% C. molybdites in boiled and digested samples could be successfully detected. The LAMP method can also be completed within 45 min, and the reaction results could be directly observed based on a color change under daylight by the naked eye. Therefore, the LAMP assay established in this study provides an accurate, sensitive, rapid, and low-cost method for the detection of C. molybdites.

Multiple authentications of high-value milk by centrifugal microfluidic chip-based real-time fluorescent LAMP.

Adulteration of food ingredients, particularly replacement of high-value milk with low-cost milk, affects food safety. For rapid and accurate identification of the possible adulterating milk species in an unknown sample, a centrifugal microfluidic chip-based real-time fluorescent multiplex loop-mediated isothermal amplification (LAMP) assay was developed to simultaneously detect milk from cow, camel, horse, goat, and yak. Using precoated primers in different reaction wells, the centrifugal microfluidic chip markedly simplified the detection process and reduced false-positive results. The entire amplification was completed within 90 min with a genomic detection limit of 0.05 ng/µL in cow, camel, horse, and goat milk and 0.005 ng/µL in yak milk. Using simulated adulterated samples for validation, the detection limit for adulterated milk samples was 2.5%, satisfying authentication requirements, as the proportion of adulterated milk higher than 10% affects economic interests. Therefore, this simple, centrifugal, microfluidic chip-based multiplex real-time fluorescent LAMP assay can simultaneously detect common milk species in commercial products to enable accurate labeling.

Application of cystourethroscopy during tracheobronchial foreign body removal in children.

Objective This study aimed to describe preliminary experiences associated with removal of tracheobronchial foreign bodies (TFBs) by cystourethroscopy (CU). Methods We performed a retrospective analysis of 127 paediatric cases of TFB removal by CU at our centre from January 2009 to August 2016. Data that were extracted from the medical records included age, sex, location and nature of the TFBs, operation time, and complications. Results All TFBs were successfully removed by CU. The mean time of the procedure was 3.38 ± 2.86 minutes. A total of 102 (80.31%) patients had successful removal of TFBs by CU during the initial trial, 19 (14.96%) were successfully treated in the second trial, and six (4.72%) required a third trial. Otolaryngologists with 2, 5, and 7 years of professional CU training showed a mean TFB removal time of 3.38 ± 2.13, 3.40 ± 3.60, and 3.37 ± 2.86 minutes, respectively. In the operations, oxygen saturation fell below 90% at an average occurrence of 0.39 times, but no patients showed a decrease below 85%. Only one patient experienced laryngeal oedema after the procedure. Conclusion CU is a useful technique and minimizes complications and operational risks during removal of paediatric TFBs.

miR-371-5p suppresses the proliferative and migratory capacity of human nasopharyngeal carcinoma by targeting BCL2.

The aim of the present study was to investigate the expression and function of microRNA (miR)-371-5p in nasopharyngeal carcinoma (NPC). The levels of miR-371-5p were analyzed in nasopharyngeal epithelium tissues, NPC tissues, human NPC cell lines and NP69 cells using reverse transcription-quantitative polymerase chain reaction analysis. The association between the level of miR-371-5p and clinicopathological variables was also investigated. Cell proliferation was determined using an MTT assay, and the activities of cell metastasis were determined using wound healing and Transwell migration assays. To assess whether miR-371-5p can combine with the targeting sequence of B-cell lymphoma 2 (BCL2) mRNA or not, a luciferase activity assay was performed. An animal experiment was used to examine the effect of miR-371-5p on the development of NPC. The results revealed that the expression of miR-371-5p was reduced in NPC samples and NPC cells. The level of miR-371-5p was associated with clinical stage and distant metastasis in patients with NPC, and was inversely associated with the protein level of BCL-2 in NPC tissues. The upregulation of miR-371-5p reduced cell growth, migration and invasion, and inhibited carcinoma growth through targeting BCL2 mRNA. Taken together, the regulation of miR-371-5p was shown to offer potential as a novel treatment approach for NPC.

Prevalence of hypertension and associated risk factors in Chinese Jing compared with Mulao populations.

OBJECTIVE: *These authors contributed equally to this work. At present, they work at the Hezhou People's Hospital, Hezhou, China.To retrospectively compare differences in the prevalence of hypertension and associated risk factors between the Chinese Jing and Mulao populations. METHODS: Subjects of Jing and Mulao ethnicities were surveyed using stratified randomized sampling. Demography, diet and lifestyle data were collected using standardized questionnaires. Several anthropometric parameters, blood pressure (BP) levels and serum lipid concentrations were obtained. RESULTS: Data from 915 Jing and 911 Mulao subjects aged ≥ 35 years were included. Diastolic BP levels and prevalence of hypertension were lower, but prevalence of isolated systolic hypertension was higher, in the Jing compared with the Mulao population. Prevalence of hypertension in the age 60-69 years, body mass index (BMI) > 24 kg/m(2), and smoker subgroups was lower in the Jing compared with the Mulao populations. Prevalence of hypertension correlated with age, cigarette smoking, triglyceride level, waist circumference, sodium intake and total dietary fibre in the Jing population; hypertension prevalence also correlated with age, triglyceride level, BMI, total fat, sodium intake and total dietary fibre in the Mulao population (unconditional logistic regression analyses). CONCLUSIONS: Prevalence of hypertension and associated risk factors were different between the two ethnic minorities, which might result from the combined effects of differences in their geographic, dietary, lifestyle, and genetic backgrounds.

Serum lipid profiles, the prevalence of dyslipidemia and the risk factors in two isolated Chinese minorities.

Both Jing and Mulao nationalities are the isolated minorities in China. Little is known about the prevalence of dyslipidemia between the two ethnic groups. Therefore, the aim of this study was to compare the differences in serum lipid profiles, the prevalence of dyslipidemia and their risk factors between the Jing and Mulao populations. A cross-sectional study of dyslipidemia was conducted in Dongxing city, Guangxi, China, during Dec 2011 and Jan 2012. A total of 1254 subjects of Jing and 1251 participants of Mulao were surveyed by a stratified randomized sampling. Information on demography, diet and lifestyle was collected with standardized questionnaire. Serum lipid levels were detected using the commercially available kits. The levels of low-density lipoprotein cholesterol (LDL-C), apolipoprotein (Apo) A1, and the ratio of ApoA1 to ApoB were lower but the levels of ApoB were higher in Jing than in Mulao (P < 0.001 for all). The prevalence of hypertriglyceridemia (32.38% vs. 24.38%), high ApoB (35.25% vs. 15.35%) and low ApoA1/ApoB ratio (22.65% vs. 16.87%) was higher and low high-density lipoprotein cholesterol (0.48% vs. 2.16%), high LDL-C (17.54% vs. 40.53%) and low ApoA1 (5.98% vs. 11.43%) was lower in Jing than in Mulao (P < 0.001 for all). The risk factors for serum lipid parameters and hyperlipidemia were different between the two ethnic groups. Serum lipid profiles, the prevalence of dyslipidemia and their risk factors are different between the Jing and Mulao populations. These differences may result from the combined effects of different diet, lifestyle, and genetic factors.