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X-linked retinitis pigmentosa (XLRP) is the most severe form of Retinitis Pigmentosa (RP) and one of the leading causes of blindness in the world. Currently, there is no effective treatment for RP. In the present study, we recruited a XLRP family and identified a 4 bp deletion mutation (c. 2234_2237del) in RPGR ORF15 with Sanger sequencing, which was located in the exact same region as the missing XES (X chromosome exome sequencing) coverage. Then, we generated cell lines harboring the identified mutation and corrected it via enhanced prime editing system (ePE). Collectively, Sanger sequencing identified a pathogenic mutation in RPGR ORF15 for XLRP which was corrected with ePE. This study provides a valuable insight for genetic counseling of the afflicted family members and prenatal diagnosis, also paves a way for applying prime editing based gene therapy in those patients.

Hydronephrosis is one of the most common diseases in urology. However, due to the difficulties in clinical trials and the lack of reliable

Insulin resistance (IR) is a typical sign of metabolic dysregulation caused by fine particulate matter (PM2.5), but the underlying signaling has not been clearly determined. Herein, a microfluidic liver-kidney microphysiological system (LK-MPS) is presented to assess the signaling pathways of IR generated by PM2.5 at 200 μg/mL for 24 h. The LK-MPS device consisted of a biomimetic liver-kidney architecture and reconstructed two circulation paths: the liver metabolism-kidney excretion (LM-KE) and kidney excretion-liver metabolism (KE-LM), by which PM2.5 is feasibly distributed in the two organs. Transmission electron microscopy (TEM) analysis revealed that PM2.5 can embed in the cytoplasm and nuclei, undergo transport by vesicles, and lead to the destruction of mitochondria. Further comprehensive immunofluorescence, enzyme-linked immunosorbent assays (ELISAs) and untargeted metabolomic analyses confirmed that PM2.5 disturbed the classic IRS-1/AKT signaling pathway (INSR, IRS-1, PI3K, AKT, GLUT2, GLUT4, and FOXO1 downregulated) and IR-related metabolic pathways: UDP-hexosamine (UDP-GlcNAc), gluconeogenesis (β-d-glucose 6-phosphate), and lipid biosynthesis (ceramide (Cer) and triacylglycerol (TG)) pathways, leading to the disorder of glucose levels. Collectively, these disorders aggravate hepatic and renal IR. Pearson's correlation coefficient test showed that elemental carbon (EC), polycyclic aromatic hydrocarbons (PAHs), and metals (Ca, Co, and V) were negatively correlated to the dysregulated proteins (INSR, IRS-1, AKT, FOXO1, GLUT2, and GLUT4). These findings may partially explain IR-related signaling pathways triggered by PM2.5.

Fine particle (PM

Background: Peroral endoscopic myotomy (POEM) is a particularly attractive intervention for achalasia. Presently, POEM has been reported to be effective and safe for achalasia in geriatric patients. Herein, this systematic review was conducted to explore the role of POEM in geriatric patients with achalasia. Method: PubMed, Embase, and Cochrane Library were searched to identify studies evaluating the clinical outcome of POEM in geriatric patients with achalasia during January 2009 to October 2020. The primary outcomes were technical and clinical success. Secondary outcomes included postoperative Eckardt score, lower esophageal sphincter (LES) pressure, adverse events, and clinical reflux. Results: There were 7 studies with a total of 469 geriatric patients, and the pooled technical success of POEM treatment was 98.1% (95% confidence interval [CI], 95.1–99.3%), and the pooled clinical success was 92.5% (95% CI, 89.3–94.8%). After POEM, the Eckardt score significantly decreased by 6.09 points (95% CI, 5.44–6.74, p < 0.00001), and the LES pressure significantly reduced by 13.53 mm Hg (95% CI, 5.14–21.91, p = 0.002). The pooled adverse events rate was 9.0% (95% CI, 4.3–17.9%), and the post-POEM clinical reflux rate was 17.4% (95% CI, 12.9–23.2%). Conclusion: Our current study demonstrated that POEM was an effective and safe technique for achalasia in geriatric patients.

Taking advantage of the diversity-oriented synthesis strategy with α,β-unsaturated carbonyl compounds, we have successfully established the DNA-compatible transformations for various heterocyclic scaffolds. The ring-closure reactions for pyrrole, pyrrolidine, pyrazole, pyrazoline, isoxazoline, pyridine, piperidine, cyclohexenone, and 5,8-dihydroimidazo[1,2-a]pyrimidine were elegantly demonstrated in a DNA-compatible format. These efforts paved the way for preparing DNA-encoded libraries with more extensive chemical space.

DNA-encoded library technology (DELT) employs DNA as a barcode to track the sequence of chemical reactions and enables the design and synthesis of libraries with billions of small molecules through combinatorial expansion. This powerful technology platform has been successfully demonstrated for hit identification and target validation for many types of diseases. As a highly integrated technology platform, DEL is capable of accelerating the translation of synthetic chemistry by using on-DNA compatible reactions or off-DNA scaffold synthesis. Herein, we report the development of a series of novel on-DNA transformations based on oxindole scaffolds for the design and synthesis of diversity-oriented DNA-encoded libraries for screening. Specifically, we have developed 1,3-dipolar cyclizations, cyclopropanations, ring-opening of reactions of aziridines and Claisen–Schmidt condensations to construct diverse oxindole derivatives. The majority of these transformations enable a diversity-oriented synthesis of DNA-encoded oxindole libraries which have been used in the successful hit identification for three protein targets. We have demonstrated that a diversified strategy for DEL synthesis could accelerate the application of synthetic chemistry for drug discovery., Constructing DNA-encoded oxindole libraries by a diversified strategy.

A mild reaction for DNA-compatible, palladium promoted Suzuki-Miyaura cross-coupling reaction of potassium Boc-protected aminomethyltrifluoroborate with DNA-conjugated aryl bromides has been developed efficiently. This novel DNA encoded chemistry reaction proceeded well with a wide range of functional group tolerance, including aryl bromides and heteroaryl bromides. Further, the utility our DNA conjugated aminomethylated arene products is demonstrated by reaction with various types of reagents (including amide formation with carboxylic acids, alkylation with aldehydes, and carbamoylation with amines) as would be desired for the production of a DNA encoded library.

Fine particulate matter (PM2.5)-induced metabolic diseases have attracted a great deal of attention recently. However, the relevant metabolic mechanisms of PM2.5 in vivo have not yet been fully described due to the lack of reliable platforms. Herein, a membrane-free liver-gut-on-chip (L-GOC) platform was developed to investigate metabolism dysregulation induced by PM2.5. A multiple organ system with a liver-gut structure and two circulation paths (L-G and G-L circulation paths) was created, and then cells were exposed to PM2.5 on this platform. Secreted high-density lipoprotein (HDL) levels were detected, which demonstrates that this multiple organ system functioned with normal physiological metabolism at the organ level. Untargeted metabolomic analysis showed that there were 364 metabolites of LO2 cells dysregulated after exposure to PM2.5 at a concentration of 200 μg/mL. Moreover, cholesterol and bile acid metabolism were significantly dysregulated. Further immunofluorescence and ELISA assays confirmed that signal transduction pathways related to cholesterol metabolism (LCAT-CE, PON1-HDL, and SRB1-HDL metabolic pathways) and bile acid metabolism (CYP7A1-CA/CDCA/DCA metabolic pathways) were disturbed. These results indicate that PM2.5 primarily disturbed cholesterol metabolism of the liver and then disrupted bile acid metabolism of the liver (primary bile acid biosynthesis) and gut (secondary bile acid biosynthesis) via related metabolic pathways. These findings may partially explain the metabolic mechanisms of cells triggered by PM2.5 exposure.

Intestinal dysbiosis contributes to the progression of renal failure and cardiovascular diseases in patients with chronic kidney disease. Probiotics is a promising intervention to improving intestinal dysbiosis. A double-blind clinical trial to investigate the ability of probiotics to modulate gut microbiota compositions in patients receiving hemodialysis (HD) was undertaken. Fifty HD patients were enrolled and randomized, receiving either probiotics or placebo for 6 months. The responses to the interventions on gut microbiome, serum and fecal metabolome, serum albumin and endotoxin, endothelial activation markers and inflammatory markers were assessed. Totally, 22 in the probiotics group (11 males; 14 non-diabetic) and 23 in the placebo group (13 males; 17 non-diabetic) completed the study. Compared to that in the placebo group, probiotics did not significantly alter species diversity of the fecal microbiome. Probiotics did, however, restore the community composition, with particular significance in non-diabetic HD patients (P = 0.007 by Adonis analysis). Specifically, according to the results of linear discriminate analysis effect size, probiotics raised the proportions of family Bacteroidaceae and Enterococcaceae, and reduced Ruminococcaceae, Halomonadaceae, Peptostreptococcaceae, Clostridiales Family XIII. Incertae Sedis and Erysipelotrichaceae in non-diabetic HD patients. Additionally, probiotics reduced the abundances of several uremic retention solutes in serum or feces, including indole-3-acetic acid-O-glucuronide, 3-guanidinopropionic acid, and 1-methylinosine (P

As a new type of environmental pollutants, micro/nano plastics (MPs/NPs) derived from plastic products are commonly contact in daily life and lead to some serious health issues. The toxicity effects of MPs/NPs on the human body have aroused wide concerns. Although MPs/NPs have been reported to be transmitted into the kidney and reproductive organs, the molecular mechanisms of MPs/NPs toxicity remain unclear due to the lack of a physiologically relevant organ-organ linking platform in vitro. Here, we present a kidney-testis microfluidic platform (KTP) with NPs exposure that enables the communication of kidney and testis chambers and reproduces endothelium-linked chambers to simulate the state in vivo. The function of KTP was assessed by cell counting kit (CCK-8), tight junction protein claudin-2 and glucose consumption. Results revealed that MPs/NPs entered the kidney and testis via endocytosis. Immunofluorescence and ELISA analysis were performed on KTP at 200 μg/mL PS-NP to identify the dysregulated proteins on cancer-related signaling pathways, including the MAPK signaling pathway (RTK, RAS, ERK, JNK, P38, NRF2, TNF-α, and TNF-α-R) and the PI3K-AKT signaling pathway (PI3K, AKT, MDM2, P53, and ΒΑD). This multi-organ platform (KTP) contributes to clarifying cancer pathways triggered by MPs/NPs exposure and provides a promising method for assessing diseases induced by environmental pollutants.

DNA-encoded library (DEL) technology provided a powerful screening platform for identifying potential bioactive small molecules with high affinity to biologically interesting targets. Essential to a successful DEL campaign are the drug-like small molecular moieties of DNA-encoded libraries with expanded chemical space. Our laboratory has been working on developing and producing novel DNA-encoded libraries that complement current reported DELs. Herein, we demonstrated a general set of DNA-compatible reactions that enable the preparation of pyrrole-based DNA-encoded libraries in which the DNA tags are linked to the N position of the pyrrole central core. Further diversification could be rapidly incorporated into the pyrrole scaffold by robust iodination and Suzuki coupling reactions.

Insulin resistance (IR) is a typical sign of metabolic dysregulation caused by fine particulate matter (PM

Mesothelin (MSLN) is considered as a potential serological tumor biomarker for early diagnosis of pancreatic cancer. Nevertheless, low sensibility, high reagent consumption, and time costs of traditional detection methods limit their utility in clinical disease diagnoses. Here, we combined the immunoassay technique with microfluidic chips to develop a microfluidic immunoassay system (MIAS) that can be used for rapid semi-quantitative detection of serum MSLN levels. The MIAS was composed of 12 uniform structures, including 12 inlets, 12 reaction chambers, and one outlet allowing measurement of four samples with three repeats, simultaneously. A unique microarray cylinder was located at the end of each reaction chamber where immunoassay was performed to trap microspheres. A feasible interception efficiency (∼80%) was attained, with microspheres filling the reaction column. It has been demonstrated that fluorescence intensity is proportional to the MSLN concentration on the MIAS (R2 = 0.95). Subsequently, 16 clinical serum samples collected from Changhai Hospital, Shanghai were selected from eight patients with pancreatic cancers, four with pancreatitis, and four with other digestive system diseases (2 gastric cancer, 1 bile duct stricture, and 1 bile duct stones). MSLN levels for these samples were detected via MIAS. The results showed a significant correlation between MIAS and traditional enzyme-linked immunosorbent assay (ELISA), with the correlation coefficient, 0.93. The detection limit of MSLN fluorescence intensity and concentration was ∼6 a.u. and ∼20 pg/mL, respectively. The entire duration of analysis by MIAS decreased to ∼40 min compared to 2 h by ELISA. Statistical analysis of MIAS data revealed that MSLN was overexpressed in pancreatic cancer than in the others (P value = 0.0014). Moreover, the diagnostic accuracies of MSLN detected by MIAS and CA19-9 detected by ELISA in hospitals were 87.5 and 81.3%, respectively. MSLN is helpful for the early diagnosis of pancreatic cancer and other diseases, and it had a significant ability to discriminate between pancreatic and nonpancreatic cancers (P value = 0.0159). The results from this study show that MIAS has the potential to become a new serological tumor marker detection platform for rapid detection and semi-quantitative determination of MSLN and would have broad applications in early clinical diagnosis.

Ambient fine particle is a crucial indicator of air pollution brought into the air by sundry natural and public events. However, a comprehensive understanding of the PM2.5-induced cytotoxicity especially the contribution of bioaerosol part is still undiscovered. Herein, an ALI microfluidics system integrated multi-omics (iTRAQ & RNA-seq) was successfully utilized to recognize the molecular mechanisms induced by microorganisms carried bioaerosol in human lung epithelial cells. The cells viability was above 98% within 21 days on this system. Moreover, the results showed that eight microorganisms-related pathways (e.g., Salmonella, amoebiasis, HTLV-1) were activated after exposure to PM2.5 for 24 h, which played a certain proportion in contributing to inflammation reaction. In addition, multi-omics demonstrated that three inflammation-related signal transduction cascades including MAPK signaling pathway, TNF signaling pathway, and TGF signaling pathway were triggered by fine particles, ultimately leading to apoptosis-related process disorder by associated cytokines like TNF, IL6, and TGF-β. Furthermore, flow cytometry analysis showed that the cell apoptosis rate increased from 3.8% to 66.7% between the cells exposed to PM2.5 (10 μg/cm2) for 24 h and untreated control cells, which indicated that the fine particles had the ability to activate apoptosis-related signal cascades and result in apoptosis. ELISA assay and western blot indicated that HO-1, JNK, IL6, TNF, NF-κB, and FGF14 were significantly increased after exposure to PM2.5 while Casp3 and FGFR were decreased, which were consistent with the multi-omics. Moreover, PM2.5 components (OC, EC, 16PAHs, As, Cu, Mn, Cl−, and NO3−) were significantly correlated to the inflammation related proteins and cytokines, which played a vital role in the inflammation and apoptosis related signaling pathways. These findings pointed to strong links among microorganisms infection, inflammation, and apoptosis in cell response to PM2.5 carried microorganisms. It also provided a new approach for understanding PM2.5-induced cytotoxicity and health risks.

Staphylococcus epidermidis infection can be a great threat after ocular surgery and can lead to post-surgery intraocular inflammation and loss of eyesight in severe cases. Existing methods of diagnosing S. epidermidis are time consuming and imprecise. In addition, using a small volume of aqueous humor samples hampers rapid detection. This study aimed to design a fast and exact method by which S. epidermidis can be detected based on a loop-mediated isothermal amplification (LAMP) assay and microfluidics. A polypod chip based on microfluidics was designed and fabricated. The chip was composed of multi layers comprising two adhesive tape layers, a pretreatment layer, a pipeline layer, and a polypod layer. It integrated the functional modules of pretreatment of the aqueous sample, self-introduction of reagents, and nucleic acid amplification in one single chip. For the LAMP assay, 11 primer sets were designed and tested to select one that resulted in high specificity and sensitivity and that could detect small amounts of nucleic acid in samples with

The demand for organic agricultural products in China is expected to have a huge increase due to the new food safety policies and the expansion of middle classes. At the same time, consumers have been increasingly concern about health and environmental issues. Organic food, an emerging market that represents healthier lifestyle, better taste and safer environment, is being recognized by people from different classes. Exploring this market from the side of consumers’ perception is rather meaningful and important. The Theory of Planned Behavior (TPB) model has been studied and applied to psychology and behaviors. Together with statistical measures, TPB and other factors can be applied to explore people’s purchasing intentions.

Fine particulate matter (PM

Background: Peroral endoscopic myotomy (POEM) has been reported to be effective in achalasia patients with prior failed endoscopic intervention (PFI). We performed this meta-analysis to compare and summarize the clinical outcome of POEM in patients with or without prior endoscopic intervention. Method: We searched relevant studies published up to March 2020. Meta-analysis for technical success, clinical success, Eckardt score, lower esophageal sphincter (LES) pressure, clinical reflux, and adverse event were conducted based on a random-effects model. Results: Eight studies enrolling 1,797 patients who underwent POEM were enrolled, including 1,128 naïve achalasia patients and 669 patients with PFI. In the PFI group, the pooled estimated rate of technical success was 97.7% (95% confidence interval [CI], 95.8–98.8%), the pooled clinical success rate was 91.0% (95% CI, 88.0–93.4%), and the pooled adverse events rate was 23.5% (95% CI, 10.6–44.1%). The Eckardt score significantly decreased by 5.95 points (95% CI, 5.50–6.40, p < 0.00001) and the LES pressure significantly reduced by 19.74 mm Hg (95% CI, 14.10–25.39, p < 0.00001) in the PFI group. There were no difference in the technical success, clinical success, and adverse events rate between the treatment-naïve group and PFI group, with a risk ratio of 1.0 (95% CI, 0.99–1.01, p = 0.89), 1.02 (95% CI, 0.98–1.06, p = 0.36), and 0.88 (95% CI, 0.67–1.16, p = 0.38), respectively. Conclusions: POEM is an effective and safe treatment for achalasia patients with prior endoscopic intervention. Randomized clinical trials are needed to further verify the efficiency and safety of the POEM in those patients.

SARS-Cov-2 has erupted across the globe, and confirmed cases of COVID-19 pose a high infection risk. Infected patients typically receive their treatment in specific isolation wards, where they are confined for at least 14 days. The virus may contaminate any surface of the room, especially frequently touched surfaces. Therefore, surface contamination in wards should be monitored for disease control and hygiene purposes. Herein, surface contamination in the ward was detected on-site using an RNA extraction-free rapid method. The whole detection process, from surface sample collection to readout of the detection results, was finished within 45 min. The nucleic acid extraction-free method requires minimal labor. More importantly, the tests were performed on-site and the results were obtained almost in real-time. The test confirmed that 31 patients contaminated seven individual sites. Among the sampled surfaces, the electrocardiogram fingertip presented a 72.7% positive rate, indicating that this surface is an important hygiene site. Meanwhile, the bedrails showed the highest correlation with other surfaces, so should be detected daily. Another surface with high contamination risk was the door handle in the bathroom. To our knowledge, we present the first on-site analysis of COVID-19 surface contamination in wards. The results and applied technique provide a potential further reference for disease control and hygiene suggestions., Graphical abstract Unlabelled Image, Highlights • First report about on-site detection of SARS-CoV-2 in wards • The whole detection process could accomplish within 45 min without nucleic acid extraction. • The presence of SARS-CoV-2 in wards was confirmed by nucleic acid isothermal amplification.

Acute toxicity assay presents vital significance in modern environmental monitoring, including online detection and in-situ assay for emergency events. Although photobacteria related detection methods were established and verified in the past decades with combination of photomultiplier tube (PMT), the price and size of PMT sensor hampered application of rapid acute toxicity assay and detection system miniaturization, especially in the resource-limited occasions. Wide application of smartphones with great low-light performance cameras could be used in photobacteria-based toxicity assay instead of the PMT methods. Herein a box-type portable detection system had been successfully established, including a disc-chip for detection, detection device, and smartphones with a high-performance camera. The system performed well showing stable temperature and rotation control. Results captured by CMOS-based camera presented a linear relationship with PMT-based detection method. An image progress algorithm was also established and tested by series diluted zinc sulfate solution as a reference substance. The system also performed well for toxicity analysis for real Atmospheric particle matter sample. The system could be used in some environmental monitoring scenarios as an alternative solution.

Air pollution has side effects on human health. Epidemiology studies indicate a positive association between ambient fine particle (PM

With the wide use of titanium dioxide nanoparticles (TiO2-NPs), the genotoxicity of TiO2-NPs, which is a factor for safety assessment, has attracted people's attention. However, their genotoxic effects in vitro remain controversial due to inconsistent reports. Therefore, a systematic review was conducted followed by a meta-analysis to reveal whether TiO2-NPs cause genotoxicity in vitro. A total of 59 studies were identified in this review through exhaustive database retrieval and exclusion. Meta-analysis results were presented based on different evaluation methods. The results showed that TiO2-NP exposure considerably increased the percentage of DNA in tail and olive tail moment in comet assay. Gene mutation assay revealed that TiO2-NPs could also induce gene mutation. However, TiO2-NP exposure had no effect on micronucleus (MN) formation in the MN assay. Subgroup analysis showed that normal cells were more vulnerable to toxicity induced by TiO2-NPs. Moreover, mixed form and small particles of TiO2-NPs increased the percentage of DNA in tail. In addition, short-term exposure could detect more DNA damage. The size, coating, duration, and concentration of TiO2-NPs influenced MN formation. This study presented that TiO2-NP exposure could cause genotoxicity in vitro. The physicochemical properties of TiO2-NPs and experimental protocols influence the genotoxic effects in vitro. Comet and gene mutation assays may be more sensitive to the detection of TiO2-NP genotoxic effects.

Influenza is a zoonotic disease, infecting a wide variety of warm-blooded animals. It is caused by an influenza virus, which has been found with hundreds of subtypes. These subtypes are often associated with different sources of infection and possess complex courses of infection. In the early stage of influenza infection, rapid subtype detection is very practicable to prevent the disease from getting worse. Herein, we presented a high-throughput microfluidic centrifugal disc for rapid detection of influenza virus subtypes. The disc realized detection reagent preloads, automated reagent control, and RT-LAMP detections. Six kinds of highly pathogenic influenza viruses could be simultaneously identified, including influenza A subtypes H1, H3, H5, H7, and H9 and influenza B virus. Two different fluorescent dyes could be used on the disc for real-time detection or read by the naked eye. The performance of the disc was demonstrated by testing the clinical samples. The integrated centrifugal disc was expected for rapid detection of influenza virus subtypes to facilitate accurate drug usage in resource-constrained settings and contribute to reduce the risk of the influenza pandemic.

The application of machine learning towards DNA encoded library (DEL) technology is lacking despite obvious synergy between these two advancing technologies. Herein, a machine learning algorithm has been developed that predicts the conversion rate for the DNA compatible reaction of a building block with a model DNA-conjugate. We exemplify the value of this technique with a challenging reaction, the Pictet-Spengler, where acidic conditions are normally required to achieve the desired cyclization between tryptophan and aldehydes to provide tryptolines. To avoid damaging the DNA our reaction conditions must be exceptionally mild, and therefore most building blocks fail to provide acceptable yields of desired product ( 78% pass rate. This is the first demonstration of using a machine learning algorithm to cull potential building blocks prior to their purchase and testing for DNA encoded library synthesis. Importantly, this allows for a challenging reaction, with an otherwise very low building block pass rate in the test reaction, to still be used in DEL synthesis. Furthermore, we discuss herein the rational design of DNA conjugated tryptophan substrates for our Pictet-Spengler reaction, and optimization of the reaction protocols. Lastly, because our protocol is solution-phase it is directly applicable to standard plate-based DEL synthesis.

Airborne fungi, including Aspergillus species, are the major causes of human asthma. Direct capture and analysis of pathogenic fungi in indoor air is important for disease prevention and control. In this paper, we demonstrated an integrated microfluidic system capable of enrichment and high-throughput detection for airborne fungal spores of Aspergillus niger, a well-known allergenic harmful species. The microfluidic system allowed semiquantitative detection of Aspergillus niger spores based on immunofluorescence analysis. To assess its contaminated level, the whole analysis time could be completed in 2-3 h including ∼1 h of enrichment and ∼1 h of target detection. The detection limit was ∼20 spores, equivalent to ∼300 spores·m-3 of the concerned targets in air. In addition, the microfluidic system has integrated sampling and sample analysis to avoid additional sample concentration step, showing the potential for point-of-care detection for other pathogenic fungal spores.

Background/Aims: Recently, the colon has been recognized as an important source of various uremic toxins in patients with end stage renal disease. Medicinal charcoal tablets are an oral adsorbent that are widely used in patients with chronic kidney disease in China to remove creatinine and urea from the colon. A parallel fecal and serum metabolomics study was performed to determine comprehensive metabolic profiles of patients receiving hemodialysis (HD). The effects of medicinal charcoal tablets on the fecal and serum metabolomes of HD patients were also investigated. Methods: Ultra-performance liquid chromatography/mass spectrometry was used to investigate the fecal and serum metabolic profiles of 20 healthy controls and 31 HD patients before and after taking medicinal charcoal tablets for 3 months. Results: There were distinct metabolic variations between the HD patients and healthy controls both in the feces and serum according to multivariate data analysis. Metabolic disturbances of alanine, aspartate and glutamate metabolism, arginine and proline metabolism figured prominently in the serum. However, in the feces, alterations of tryptophan metabolism, lysine degradation and beta-alanine metabolism were pronounced, and the levels of several amino acids (leucine, phenylalanine, lysine, histidine, methionine, tyrosine, and tryptophan) were increased dramatically. Nineteen fecal metabolites and 21 serum metabolites were also identified as biomarkers that contributed to the metabolic differences. Additionally, medicinal charcoal treatment generally enabled the serum and fecal metabolomes of the HD patients to draw close to those of the control subjects, especially the serum metabolic profile. Conclusion: Parallel fecal and serum metabolomics uncovered the systematic metabolic variations of HD patients, especially disturbances in amino acid metabolism in the colon. Medicinal charcoal tablets had an impact on the serum and fecal metabolomes of HD patients, but their exact effects still need to be studied further.

Different microfluidic design or function modules are the key for the microfluidic chip to fullfill various functions. We herein report a simple microfilter module that conveniently integrates semiporous membranes into polydimethylsiloxane (PDMS) substrates with leakage-free bonding, which achieves the efficient enrichment of airborne and waterborne pathogens on microfluidic chips. The microfilter is able to rapidly enrich and concentrate ∼10−1 μm-scale particles with high efficiency due to the complementary advantages of microfluidics (microliter-scale fluid control) and the nature of polyester (or polycarbonate) membranes (high precision and density of pores, and high hydrophilicity). Fabrication can be conveniently performed by effective loading control of the sealing agent. For further studies on different pathogen concentrations and detections, the module was integrated with loop-mediated isothermal amplification (LAMP) on a PDMS chip for enrichment and visual analysis of pathogens from body fluid samples with limited volume and low concentration. It was also capable of collection and in-situ immunofluorescence detection of parasite oocysts (cysts) in drinking water with recovery rates of ∼90%. Besides, by integration of the microfilter and a LAMP chip on a portable device, we successfully established a point-of-care system for enrichment and analysis of airborne pathogens.

An increasing number of investigations have revealed that podocytes play a crucial role in the development and progression of diabetic nephropathy (DN). Quercetin (2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one) is the most common flavonol in the diet and is one of the most prominent dietary antioxidants, which might have a protective effect on DN. The present study was designed to investigate the protective effect of quercetin on podocyte impairment in a rat model of DN, as well as underlying molecular mechanisms. All diabetic rats were induced by a single intraperitoneal injection of streptozotocin, and quercetin was administered daily at a dose of 50 mg kg−1 or 100 mg kg−1 for 12 weeks. In the present study, quercetin markedly decreased blood glucose levels, kidney-to-body weight ratio, albuminuria, creatinine clearance rate, blood urea nitrogen, and triglycerides and significantly attenuated oxidative stress. Moreover, quercetin was observed to inhibit podocyte effacement and decrease the thickness of glomerular basement membranes. Mechanistically, quercetin significantly increased the expression of podocyte-specific markers nephrin and podocin and decreased expression of the podocyte injury marker desmin in DN rats. Quercetin also inhibited activation of the TGF-β1/Smad signaling pathway in DN rats by decreasing expression of TGF-β1, p-Smad2, and p-Smad3, and increasing Smad7 expression. These findings suggest that quercetin administration ameliorated podocyte injury in DN rats, possibly by inhibiting oxidative stress and the TGF-β1/Smad signaling pathway. Thus, quercetin may be manipulated to act as a potential drug for prevention of early diabetic nephropathy.

Here we presented a novel point-of-care detection method for semi-quantitative assessments of specific pathogenic bioaerosols based on the portable microfluidics-based device. Sample enrichment, DNA amplification, and detection were performed sequentially on the device with simple procedures, perfect for on-site application. The methods for the assessments of four pathogenic bioaerosol biomarkers, including Staphylococcus aureus , Pseudomonas aeruginosa , Klebsiella pneumonia , and Acinetobacter baumannii , were respectively established that inclusive of the selections of specific primers, optimizations of on-chip DNA extractions, and the establishments of semi-quantitative curves. Then the methods were verified by the assessments of generated mimic bioaerosols. The results demonstrated that pathogen quantities could be estimated by using the method to reflect airborne dissemination. The presented method could provide valuable data for the forecasting pathogen spreading risks in high populated urban area and transportation hub, and potentially for the requirements of epidemiologic studies on dose-response estimates.

Human exposure to PM2.5 causes several adverse health effects. Skin is the first barrier against harmful environmental substances and can directly contact with PM2.5, but there is no study about PM2.5-induced cytotoxicity in human skin cells on the molecular level partially due to the shortcomings of traditional research methods. In present study, we established a microfluidic system including a cell culture chip integrated with a high-throughput protein microarray chip to investigate the mechanism of PM2.5-mediated cytotoxicity in human HaCaT cells. We found that PM2.5 was lodged inside the cytoplasm, mitochondria and nucleus of HaCaT cells by TEM. Flow cytometry analysis indicated that the cell apoptosis rate increased from 0.49% to 53.4%. The results of protein microarray showed that NF-κB and NALP3 signal transductions were activated in HaCaT cells after PM2.5 stimulations, up-regulating the expression of IL-1β and IL-6, which resulted in inflammatory response in HaCaT cells. Our findings provide a molecular insight into PM2.5-induced skin injury.

PM2.5 (particulate matter less than 2.5 μm in diameter) is considered as a harmful carcinogen. Determining the precise relationship between the chemical constituents of PM2.5 in the air and cancer progression could aid the treatment of environment related disease and establishing risk reduction strategies. Herein, we used transcriptomics (RNA-seq) and an integrated microfluidic system to identify the global gene expression and differential target proteins expression induced by ambient fine particles collected from the heavy haze in China. The results clearly indicated that cancer related pathways exhibited the strongest dysregulation. The ambient fine particles could be uptaken into the cells by pinocytosis, mainly promoting the PI3K-Akt pathway, FGF/FGFR/MAPK/VEGF signaling, and the JAK-STAT pathway, leading to evading apoptosis, sustained angiogenesis, and cell proliferation, which are the most important hallmarks of cancer. And fine particles also have been demonstrated to create intracellular reactive...

Homocysteine (Hcy) level characterizes a progressive increase in chronic kidney disease (CKD). In fact, Hcy accumulation is considered to be a crucial biochemical culprit in CKD progression, but the mechanism underlying this remains poorly understood. This study investigated the role of Hcy in glomerular mesangial cell (MC) apoptosis and the potential involvement of autophagy and endoplasmic reticulum (ER) stress in this process, shedding light on Hcy toxicity in kidney disease. Human mesangial cells (HMCs) were incubated with different concentrations of Hcy for different times. Flow cytometry was used to determine the proportion of apoptotic cells and western blotting was used to analyze protein levels after the administration of Hcy, endoplasmic reticulum inhibitor 4-phenylbutyric acid (4-PBA), and Atg5 siRNA. The results demonstrated that the cell viability gradually decreased and the proportion of HMCs undergoing apoptosis increased with increasing Hcy concentration and prolonged incubation time. Meanwhile, levels of the apoptosis-related proteins Bax and cleaved caspase-3 were significantly increased, while ER stress-related proteins such as ATF4, CHOP, GRP78, and phospho-eIF2α significantly increased. Levels of cleaved LC3, and beclin1 and Atg5 proteins also increased, accompanied by p62 degradation, indicating autophagy activation. 4-PBA effectively inhibited ER stress and reversed Hcy-induced apoptosis and autophagy. Moreover, Atg5 siRNA alleviated Hcy-induced apoptosis. Taken together, these results suggest that Hcy induces HMC apoptosis in a dose- and time-dependent manner

Fine particulate matter less than 2.5 μm in diameter (PM2.5) is regarded as a carcinogenic factor, but the mechanism has been left unexplored. Our goal was to reveal the carcinogenic mechanism at the gene and protein level under the inhalational air–liquid interface (ALI) condition. Herein, we developed an ALI organ-level lung microfluidic platform (ALI-OLMP) carrying lung epithelial cell line BEAS-2B and human pulmonary microvascular endothelial cells (HPMEC); the cell viability was above 98% within 14 days on this system, which was used to mimic the practical alveolar microenvironment for the multiomics analysis, to identify the global gene and protein expression after exposure to PM2.5 in Shanghai, China from 2014 to 2015. The combined RNA-Seq and iTRAQ analysis indicated that the unique set was 2532 genes at 10 μg/cm2 of PM2.5, and there were also at least 25 identical activated signal transduction cascades including bladder cancer, transcriptional dysregulation in cancer, the TP53 (p53) signaling pat...

Air pollution has side effects on human health. Epidemiology studies indicate a positive association between ambient fine particle (PM2.5, or particles less than 2.5 μm in diameter) concentration and lung cancer. However, how fine particles affect lung cancer at the molecular level and related therapeutic methods to address these diseases are unclear. Here, the multi-omics analysis (DNA methylation and transcriptomic) was used to detect human bronchial epithelial cells (HBE), that were exposed to PM2.5 using a quantified, small, portable, and organ-level air-liquid interface microfluidic system that mimics lung functions. The results indicate that 36,838 differentially methylated genes were detected. Of these 33,796 genes were hypomethylated (beta 0). RNA-Seq analysis demonstrated that 19,489 genes were upregulated (log2FC > 0), and 16,659 were downregulated. Furthermore, the calcium and apoptosis pathways were activated according to multi-omics analysis. The change in EGFR gene expression after PM2.5 exposure was the result of alterations of the cellular DNA methylome in the promoter. Inhibition or down-regulation of EGFR could result in the regulation of the downstream intracellular Ca2+ concentration and apoptosis via the EGFR/PLCγ and EGFR/STAT/Bcl-XL pathways after PM2.5 exposure. EGFR inhibitors decrease the Ca2+ concentration of cells, thereby strengthening the effects of fine particles on apoptosis. In short, the Ca2+ concentration and the apoptosis of cells can be regulated via EGFR related pathway after PM2.5 exposure. The EGFR may be a potentially promising therapeutic target for the treatment of air pollution-induced lung cancer through regulation of the intracellular Ca2+ concentration and apoptosis.

In recent years, green tides caused by water eutrophication, has brought serious environmental problems. Enteromorpha prolifera (E. prolifera), an opportunistic macroalgae, is one of the main source contributing to the formation of green tides. It has been estimated that the excessive growth of E. prolifera is closely related to various reproductive ways of germ cells which are at the micrometer scale. Here we report a microfluidic device named Germ Cell Capture Chip (GCChip) to investigate the E. prolifera reproductive mechanism. GCChip integrates the functions of algal growing, and the release, capture and selective culture of germ cells. Automatic separation and capture of germ cells on the chip allows to study germ cells’ response to different stimuli. The novel device greatly facilitates long-term live-cell imaging at cellular resolution and implements the rapid and accurate exchange of growth medium without manual intervention. Results showed that the starting time of germ cell releases were earlier on the chip than that of traditional experiments with more concentrated breakout. Moreover, GCChip can be widely applied on the study of other algae. The study of algae growth process, including the elongation of somatic cell, the generation, and the release of reproductive cells, can all be improved by using this microfluidic platform.

Circulating tumor cells (CTCs) have been proposed to be an active source of metastasis or recurrence of hepatocellular carcinoma (HCC). The enumeration and characterization of CTCs has important clinical significance in recurrence prediction and treatment monitoring in HCC patients. We previously developed a unique method to separate HCC CTCs based on the interaction of the asialoglycoprotein receptor (ASGPR) expressed on their membranes with its ligand. The current study applied the ligand-receptor binding assay to a CTC-chip in a microfluidic device. Efficient capture of HCC CTCs originates from the small dimensions of microfluidic channels and enhanced local topographic interactions between the microfluidic channel and extracellular extensions. With the optimized conditions, a capture yield reached > 85% for artificial CTC blood samples. Clinical utility of the system was further validated. CTCs were detected in all the examined 36 patients with HCC, with an average of 14 ± 10/2 mL. On the contrary, no CTCs were detected in healthy, benign liver disease or non-HCC cancer subjects. The current study also successfully demonstrated that the captured CTCs on our CTC-chip were readily released with ethylene diamine tetraacetic acid (EDTA); released CTCs remained alive and could be expanded to form a spheroid-like structure in a 3-dimensional cell culture assay; furthermore, sensitivity of released CTCs to chemotherapeutic agents (sorafenib or oxaliplatin) could be effectively tested utilizing this culture assay. In conclusion, the methodologies presented here offer great promise for accurate enumeration and easy release of captured CTCs, and released CTCs could be cultured for further functional studies.