Single-lens multi-mirror laser stereo vision-based system for measuring internal thread geometrical parameters.

Oilfield pipes with out-of-tolerance internal thread can lead to failures, so the internal thread geometric parameters need to be measured. To tackle the problem of the low efficiency, poor accuracy, easy wear, and poor accessibility of existing methods, a single-lens multi-mirror laser stereo vision-based system for measuring geometric parameters of the internal thread is proposed, which allows the measurement of three parameters in one setup by completely reproducing the three-dimensional (3D) tooth profiles of the internal thread. In the system design, to overcome the incomplete representation of imaging parameters caused by insufficient consideration of dimensions and structural parameters of the existing models, an explicit 3D optical path model without a reflecting prism is first proposed. Then, considering the intervention of the reflecting prism, a calculation model for the suitable prism size and the final imaging parameters of the vision system is proposed, which ensures the measurement accessibility and accuracy by solving the problem that the existing system design only depends on experience without theoretical basis. Finally, based on the American Petroleum Institute standard, internal thread geometric parameters are obtained from the vision-reconstructed 3D tooth profiles. According to the optimized structural parameters, a vision system is built for measuring the internal thread geometric parameters of two types of oilfield pipes. Accuracy verification and typical internal thread measurement results show that the average measurement errors of the vision system proposed for the pitch, taper, and tooth height are 0.0051 mm, 0.6055 mm/m, and 0.0071 mm, respectively. Combined with the vision measurement time of 0.5 s for the three parameters, the above results comprehensively verify the high accuracy and high efficiency of the vision-based system.

Vision measurement system for geometric parameters of tubing internal thread based on double-mirrored structured light.

Geometric parameter measurement of tubing internal thread is critical for oil pipeline safety. In response to the shortcomings of existing methods for measuring internal thread geometric parameters, such as low efficiency, poor accuracy, and poor accessibility, this paper proposes a vision system for measuring internal thread geometric parameters based on double-mirrored structured light. Compared to previous methods, our system can completely reproduce the internal thread tooth profiles and allows multi-parameter measurement in one setup. To establish the correlation between the structural and imaging parameters of the vision system, three-dimensional (3D) optical path models (OPMs) for the vision system considering the mirror effect of the prism is proposed, which extends the scope of the optical path analysis and provides a theoretical foundation for designing the structural parameters of the vision system. Moreover, modeling and three-step calibration methods for the vision system are proposed to realize high-accuracy restoration from the two-dimensional (2D) virtual image to the actual 3D tooth profiles. Finally, a vision measurement system is developed, and experiments are carried out to verify the accuracy and measure the three geometric parameters (i.e., taper, pitch, and tooth height) of typical internal threads. Based on the validation results using the reference system, the vision measurement accuracy and efficiency are 6.7 and 120 times that of the traditional system, which verifies the measurement effectiveness and accuracy of the vision system proposed in this paper.

Surface-Enhanced Raman Spectroscopy for Bioanalysis: Reliability and Challenges.

Surface-enhanced Raman spectroscopy (SERS) inherits the rich chemical fingerprint information on Raman spectroscopy and gains sensitivity by plasmon-enhanced excitation and scattering. In particular, most Raman peaks have a narrow width suitable for multiplex analysis, and the measurements can be conveniently made under ambient and aqueous conditions. These merits make SERS a very promising technique for studying complex biological systems, and SERS has attracted increasing interest in biorelated analysis. However, there are still great challenges that need to be addressed until it can be widely accepted by the biorelated communities, answer interesting biological questions, and solve fatal clinical problems. SERS applications in bioanalysis involve the complex interactions of plasmonic nanomaterials with biological systems and their environments. The reliability becomes the key issue of bioanalytical SERS in order to extract meaningful information from SERS data. This review provides a comprehensive overview of bioanalytical SERS with the main focus on the reliability issue. We first introduce the mechanism of SERS to guide the design of reliable SERS experiments with high detection sensitivity. We then introduce the current understanding of the interaction of nanomaterials with biological systems, mainly living cells, to guide the design of functionalized SERS nanoparticles for target detection. We further introduce the current status of label-free (direct) and labeled (indirect) SERS detections, for systems from biomolecules, to pathogens, to living cells, and we discuss the potential interferences from experimental design, measurement conditions, and data analysis. In the end, we give an outlook of the key challenges in bioanalytical SERS, including reproducibility, sensitivity, and spatial and time resolution.

[Correlation of DEFB1 polymorphisms with chronic periodontitis in Henan Han population].

OBJECTIVE: To investigate the association between the polymorphisms of 5'-UTR -52G/A (rs1799946), -44C/G (rs1800972), -20G/A (rs11362) in DEFB1 gene with chronic periodontitis in Henan Han population. METHODS: Peripheral blood genomic DNA of 436 patients with chronic periodontitis and 440 healthy controls were extracted and subjected to PCR-Sanger sequencing to determine the genotypes of DEFB1 5'-UTR -52G/A (rs1799946), -44C/G (rs1800972) and -20G/A (rs11362). The distribution of genotypes, allele frequencies and risk factors were analyzed by chi-square test and Logistic regression. RESULTS: There was no significant difference between healthy controls and chronic periodontitis in the genotype of -52G/A PCR- (rs1799946) and -20G/A (rs11362) (P> 0.05). While a significant difference was found between healthy controls and chronic periodontitis in -44C/G (rs1800972), the CC and CG genotype rate in the two groups were 64.5%, 82.1% and 28.2%, 14.4% respectively. One-way logistic analysis showed that the CG, GG genotype and allele G might be a protective factor. CONCLUSION: The DEFB1 -44C/G (rs1800972) is associated with chronic periodontitis in Henan Han population, and the -44CG, GG genotype and G allele may be the protective factors of chronic periodontitis in Henan Han population.

Recent Advances in Cyclodextrin-Based Light-Responsive Supramolecular Systems.

Cyclodextrins (CDs), one of the host molecules in supramolecular chemistry, can host guest molecules to form inclusion complexes via non-covalent and reversible host-guest interactions. CD-based light-responsive supramolecular systems are typically constructed using CDs and guest molecules with light-responsive moieties, including azobenzene, arylazopyrazole, o-nitrobenzyl ester, pyrenylmethyl ester, coumarin, and anthracene. To date, numerous efforts have been reported on the topic of CD-based light-responsive supramolecular systems, but these have not yet been highlighted in a separated review. This review summarizes the efforts reported over the past ten years. The main text of this review is divided into five sections (vesicles, micelles, gels, capturers, and nanovalves) according to the formation of self-assemblies. This feature article aims to afford a comprehensive understanding of the light-responsive moieties used in the construction of CD-based light-responsive supramolecular systems and to provide a helpful guide for the further design of CD-based light-responsive supramolecular systems.

High flavonoid accompanied with high starch accumulation triggered by nutrient starvation in bioenergy crop duckweed (Landoltia punctata).

BACKGROUND: As the fastest growing plant, duckweed can thrive on anthropogenic wastewater. The purple-backed duckweed, Landoltia punctata, is rich in starch and flavonoids. However, the molecular biological basis of high flavonoid and low lignin content remains largely unknown, as does the best method to combine nutrients removed from sewage and the utilization value improvement of duckweed biomass. RESULTS: A combined omics study was performed to investigate the biosynthesis of flavonoid and the metabolic flux changes in L. punctata grown in different culture medium. Phenylalanine metabolism related transcripts were identified and carefully analyzed. Expression quantification results showed that most of the flavonoid biosynthetic transcripts were relatively highly expressed, while most lignin-related transcripts were poorly expressed or failed to be detected by iTRAQ based proteomic analyses. This explains why duckweed has a much lower lignin percentage and higher flavonoid content than most other plants. Growing in distilled water, expression of most flavonoid-related transcripts were increased, while most were decreased in uniconazole treated L. punctata (1/6 × Hoagland + 800 mg•L-1 uniconazole). When L. punctata was cultivated in full nutrient medium (1/6 × Hoagland), more than half of these transcripts were increased, however others were suppressed. Metabolome results showed that a total of 20 flavonoid compounds were separated by HPLC in L. punctata grown in uniconazole and full nutrient medium. The quantities of all 20 compounds were decreased by uniconazole, while 11 were increased and 6 decreased when grown in full nutrient medium. Nutrient starvation resulted in an obvious purple accumulation on the underside of each frond. CONCLUSIONS: The high flavonoid and low lignin content of L. punctata appears to be predominantly caused by the flavonoid-directed metabolic flux. Nutrient starvation is the best option to obtain high starch and flavonoid accumulation simultaneously in a short time for biofuels fermentation and natural products isolation.

The Efficacy of Liposomal Bupivacaine Using Periarticular Injection in Total Knee Arthroplasty: A Systematic Review and Meta-Analysis.

BACKGROUND: Total knee arthroplasty (TKA) is gradually emerging as the treatment of choice for end-stage osteoarthritis. In the past, the method of liposomal bupivacaine by periarticular injection (PAI) showed better effects on pain reduction and opioid consumption after surgery. However, some recent studies have reported that liposomal bupivacaine by PAI did not improve pain control and functional recovery in patients undergoing TKA. Therefore, this meta-analysis was conducted to determine whether liposomal bupivacaine provides better pain relief and functional recovery after TKA. METHODS: Web of Science, PubMed, Embase, and the Cochrane Library were comprehensively searched. Randomized controlled trials, controlled clinical trials, and cohort studies were included in our meta-analysis. Eleven studies that compared liposomal bupivacaine using the PAI technique with the conventional PAI method were included in our meta-analysis. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines and Cochrane Handbook were applied to assess the quality of the results published in all included studies to ensure that the results of our meta-analysis were reliable and veritable. RESULTS: Our pooled data analysis demonstrated that liposomal bupivacaine was as effective as the control group in terms of visual analog scale score at 24 hours (P = .46), 48 hours (P = .43), 72 hours (P = .21), total amount of opioid consumption (P = .25), range of motion (P = .28), length of hospital stay (P = .53), postoperative nausea (P = .34), and ambulation distance (P = .07). CONCLUSION: Compared with the conventional PAI method, liposomal bupivacaine shows similar pain control and functional recovery after TKA. Considering the cost for pain control, liposomal bupivacaine is not worthy of being recommended as a long-acting alternative analgesic agent using the PAI method.

Ordered carbon nanotubes-n-dodecyl tetraethylene monoether liquid crystal composites through phase separation induced by poly(ethylene glycol).

Carbon nanotubes (CNTs) were incorporated into a lyotropic liquid crystal (LLC) matrix at room temperature through spontaneous phase separation. The phase separation process occurred in n-dodecyl tetraethylene monoether (C12E4) solutions induced by the hydrophilic polymer, poly(ethylene glycol) (PEG). It was found that the molecular weight of PEG has a significant effect on the CNTs-C12E4 system, which not only influences the phase behavior of the system but also changes the properties of the CNTs-LLC composites. Polarized optical microscopy (POM) images, combined with small-angle X-ray scattering (SAXS) results, indicate that CNTs incorporate within the layers of the lamellar LLCs without destroying the structure of LLCs. Moreover, UV-vis absorption, Raman spectra and rheological measurements were performed to investigate the characteristic properties of the CNTs-LLC composites. This study not only gives a more comprehensive understanding of polymer-induced phase separation, but also expands the potential uses of CNTs-LLC composites in nanotechnology.

Systemic lupus erythematosus combined with Castleman disease and secondary paraneoplastic pemphigus: a case report.

BACKGROUND: The literature describes a case of systemic lupus erythematosus (SLE) complicated with Castleman's disease (CD) and secondary paraneoplastic pemphigus (PNP). CASE PRESENTATION: A 12-year-old female presented with a neck mass, rash, arthralgia, and skin and mouth ulceration for 5 years were admitted. All blood cells were low. Multiple autoantibodies associated with SLE were positive. The pathology of the neck mass revealed the classical manifestations of CD. She was treated with prednisone, hydroxychloroquine, leflunomide, thalidomide, and dressings. Pathological examination of the skin revealed PNP. The neck mass was removed and continued to take antirheumatic drugs. At subsequent follow-up, the patient's disease status was stable and the skin mucosal lesion did not recur. CONCLUSION: The case of simultaneous SLE, CD, and PNP in children was rarely reported, and the correct diagnosis of the disease will help to take timely treatment.

Eco-friendly bio-encapsulation from sodium alginate-trehalose-kaolin and its performance evaluation in improving plant growth under salt or/and drought conditions.

Plant growth-promoting bacterium plays a significant role in improving plant tolerance to abiotic stresses. However, there are low survival and poor effect in field application, especially in unfavorable environments. Our previous study suggested that encapsulation of Bacillus pumilus G5 from polyvinyl alcohol­sodium alginate could improve plant growth and soil fertility under drought and salt soil conditions. However, in the G5 microbeads, the polyvinyl alcohol could not be degraded after entering the soil, and the loss of viable bacteria was severe during the drying process. Achieving a more eco-friendly and efficient formulation based on biodegradable polymers can have significant effects on increasing the quantity and quality of agricultural products. Herein, G5 has immobilized in the composite wall of sodium alginate-trehalose-kaolin microbeads and then evaluated the performance, and applied on the Pharbitis nil under salt or/and drought stress by pot experiment. A 2 % sodium alginate, 1 % trehalose, and 1 % kaolin formulation for the coating films resulted in optimal G5 microbeads embedding efficiency, viable bacteria, degradation rate, and sustained release. Also, the G5 microbeads exhibited longer storage life than that of the G5 suspension. Scanning electron microscopy revealed that the G5 microcapsules had a near-spherical structure with a particle size of around 1000 µm forming a continuous dense composite wall membrane with obvious protrusions and folds on the surface, which facilitated the release of the G5 strain. The interior of the G5 capsule was rough and suitable for bacterial attachment. Infrared spectroscopy showed that the G5 microcapsules are a simple physical mixture with no chemical reaction between the excipients, making the G5 microcapsules chemically stable. The inclusion of the G5 microcapsules considerably induced Pharbitis nil seedlings growth and biomass under drought and/or salt stress. In the rhizosphere soil of Pharbitis nil, the G5 microcapsules increased the total cultivable bacteria population, the activities of invertase, urease, phosphatase, and catalase, and the contents of available nitrogen and available phosphorus. We concluded that a suitable formulation by bio-encapsulation with eco-friendly excipients for alleviating drought and/or salt stress in plants will be advantageous in sustainable agriculture.

Study on the antioxidative mechanism of tocopherol loaded ethyl cellulose particles in thermal-oxidized soybean oil.

Our previous study proposed preparation method of tocopherol (Toc) loaded ethyl cellulose (EC) particles as antioxidant due to instability of Toc under high temperature. The present study aimed to explore the antioxidant mechanism of loaded particles. Results showed that loaded particles prepared by EC of different viscosities (EC9, EC70, EC200) had antioxidative effect, and the antioxidant activity increased with EC viscosity. Fourier transform infrared analysis demonstrated that the interaction between EC and tocopherol was mainly hydrogen bond. Loaded particles retained effectively the thermal degradation of Toc and thus enhanced the antioxidant activity. Further investigation into thermal oxidation of EC inferred the possible antioxidative mechanism included two aspects. One was that Toc was fixed in the network structure of loaded particles formed by EC to provide a barrier for avoiding degradation. Another was that EC and Toc acted on different stages of lipid oxidation, playing the antioxidative effect together.

Encapsulation of Bacillus pumilus G5 from polyvinyl alcohol­sodium alginate (PVA-SA) and its implications in improving plant growth and soil fertility under drought and salt soil conditions.

Drought and salt stresses adversely affect the growth and yield of plants in agricultural production. Bacillus pumilus, an important plant growth-promoting bacterium, play a significant role in improving plant tolerance to abiotic stresses. In this study, B. pumilus G5 were immobilized in polyvinyl alcohol­sodium alginate (PVA-SA) microbeads and then applied on the Pharbitis nil under drought and salt stresses by pot experiment. Orthogonal array experiments showed that the optimal immobilization conditions of PVA-SA immobilized G5 microbeads were adsorbent 6.0%, PVA: SA 1:1 (3.0%), CaCl2 4.0%, and bacterium: embedding agent (PVA-SA) 3:4; And the G5 microbeads produced at the optimal condition exhibited better cultivable bacteria count, encapsulation rate, expansion rate and mechanical strength. Pot experiment showed that G5 microbeads significantly increased the length and diameter of root and stem, and dry weight of P. nil during experimental stage under drought and salt stress. G5 microbeads also increased the total cultivable bacteria population, the activities of invertase (INV), urease (URE), phosphatase (PHO) and catalase (CAT), and the contents of available nitrogen (AN) and available phosphorus (AP) in the rhizosphere soil of P. nil. Therefore, our study obtained the optimal process of G5 microbeads, and confirmed its effect on improved plant growth and soil chemical and biological properties of P. nil. Thus it can be used as sustainable tool for eco-friendly bio-inoculants at salinity soil within arid and semi-arid areas.

Characterization of cold atmospheric plasma-modified dentin collagen.

The aim of this study was to evaluate the crosslinking effect of the radio-frequency atmospheric-pressure glow discharge (RF-APGD) plasma jet treatment on dentin collagen. The dentin collagen was treated by an RF-APGD plasma jet with the gas temperature of 4°C under different treatment times, while the control was a non-treatment group. The dentin collagen was characterized in terms of atomic force microscopy-based nanoindentation, differential scanning calorimeter, Raman analysis and X-ray photoelectron spectroscopy (XPS) measurement. The crosslinking effect of the plasma-treated dentin collagen was found compared to that of the control group. The elastic modulus and denaturation temperature of the dentin collagen after plasma treatment for 30 s were significantly higher than those in the control group (p<0.05). The RF-APGD plasma jet treatment can promote the crosslinking of the dentin collagen, which is of great significance to improve its mechanical and thermal stabilities.

Non-targeted analysis and risk assessment of non-volatile compounds in polyamide food contact materials.

Small molecules in food contact materials may migrate into food during their contact. To extensively analyze the migrants, non-targeted screening is needed to detect the migrants. The migrants' detection is difficult because of the complexity and the trace amount of the migrants. In this work, the dissolution precipitation method was used to extract small molecules in Polyamide (PA) kitchenware. The extract solutions were screened by ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-QTOF-MS) for non-targeted analysis and 64 different small molecules in materials were identified through the screening of a self-built database. Then, migration tests were performed to analyze migrants in food simulants. It suggests that the abundance of PA oligomers was the highest in migrants. The risk assessment of migrants revealed that the exposure of most migrants was at a safer level unlike the exposure of PA oligomers that exceeded their threshold of toxicological concern (TTC).

Naringin-inlaid silk fibroin/hydroxyapatite scaffold enhances human umbilical cord-derived mesenchymal stem cell-based bone regeneration.

OBJECTIVES: Large bone defects are a common, debilitating clinical condition that have substantial global health and economic burden. Bone tissue engineering technology has become one of the most promising approaches for regenerating defective bones. In this study, we fabricated a naringin-inlaid composite silk fibroin/hydroxyapatite (NG/SF/HAp) scaffold to repair bone defects. MATERIALS AND METHODS: The salt-leaching technology was used to fabricate the NG/SF/HAp scaffold. The cytocompatibility of the NG/SF/HAp scaffold was assessed using scanning electron microscopy, live/dead cell staining and phalloidin staining. The osteogenic and angiogenic properties were assessed in vitro and in vivo. RESULTS: The porous NG/SF/HAp scaffold had a well-designed biomimetic porous structure with osteoinductive and angiogenic activities. A gene microarray identified 854 differentially expressed genes between human umbilical cord-derived mesenchymal stem cells (hUCMSCs) cultured on SF-nHAp scaffolds and cells cultured on NG/SF/HAp scaffolds. The underlying osteoblastic mechanism was investigated using hUCMSCs in vitro. Naringin facilitated hUCMSC ingrowth into the SF/HAp scaffold and promoted osteogenic differentiation. The osteogenic and angiogenic capabilities of cells cultured in the NG/SF/HAp scaffold were superior to those of cells cultured in the SF/HAp scaffold. CONCLUSIONS: The data indicate the potential of the SF/HAp composite scaffold incorporating naringin for bone regeneration.

Biodegradable materials for bone defect repair.

Compared with non-degradable materials, biodegradable biomaterials play an increasingly important role in the repairing of severe bone defects, and have attracted extensive attention from researchers. In the treatment of bone defects, scaffolds made of biodegradable materials can provide a crawling bridge for new bone tissue in the gap and a platform for cells and growth factors to play a physiological role, which will eventually be degraded and absorbed in the body and be replaced by the new bone tissue. Traditional biodegradable materials include polymers, ceramics and metals, which have been used in bone defect repairing for many years. Although these materials have more or fewer shortcomings, they are still the cornerstone of our development of a new generation of degradable materials. With the rapid development of modern science and technology, in the twenty-first century, more and more kinds of new biodegradable materials emerge in endlessly, such as new intelligent micro-nano materials and cell-based products. At the same time, there are many new fabrication technologies of improving biodegradable materials, such as modular fabrication, 3D and 4D printing, interface reinforcement and nanotechnology. This review will introduce various kinds of biodegradable materials commonly used in bone defect repairing, especially the newly emerging materials and their fabrication technology in recent years, and look forward to the future research direction, hoping to provide researchers in the field with some inspiration and reference.

Efficacy and safety of repetitive transcranial magnetic stimulation for peripartum depression: A meta-analysis of randomized controlled trials.

This study aims to summarize and systematically review the efficacy and safety of repetitive transcranial magnetic stimulation (rTMS) for women with peripartum depression (PPD). Several databases (Wanfang, CNKI, VIP, CBM, PubMed, Embase, Cochrane Library, PsyINFO, Web of Science, and Clinical trial) were searched from inception until April 12, 2020. In total, ten randomized controlled trials (RCTs) met the eligibility criteria and were included in this systematic review. We calculated the combined effect size (standardized mean difference [SMD] and odds ratio [OR]) for the corresponding effects models. The aggregated result of 10 trials indicated a significant benefit of rTMS on PPD, and the aggregated result of remission showed significantly positive effects of Test group VS. Control group. In terms of treatment adverse effects, the aggregated result showed no statistical significance of headache and dry mouth between the two groups. The results of the meta-analysis suggest that rTMS is an effective and safe intervention for PPD. Owing to poor methodological quality among the included studies, high-quality multicenter RCTs are needed to further verify the effects of this treatment.

Construction of EVA/chitosan based PEG-PCL micelles nanocomposite films with controlled release of iprodione and its application in pre-harvest treatment of grapes.

A novel nanocomposite poly(ethylene-co-vinyl acetate) (EVA) film with controlled in vitro release of iprodione (ID) was prepared. Chitosan (CS) was used as the reinforcement which enhances the water and oxygen permeability of films. ID loaded poly(ethylene glycol)-poly(ε-caprolactone) (PEG-PCL) (IPP) micelles were used as the drug carrier which endows the films with antifungal and controlled release ability. IPP micelles with spherical shape and uniform size were obtained, and the maximum encapsulation efficacy (EE) was 91.17 ± 5.03% by well controlling the feeding amount of ID. Incorporation CS could improve the oxygen and moisture permeability of films, and the maximum oxygen permeability (OP) and water vapor transmission rate (WVTR) were 477.84 ± 13.03 cc/(m2·d·0.1 MPa) and 8.60 ± 0.25 g m-2 d-1, respectively. After loading IPP micelles, the films showed an improved antifungal ability and temperature-sensitive drug release behavior, and were found to enhance the quality of grapes by pre-harvest spraying.

Hyperbranched poly(ß-amino ester) based polyplex nanopaticles for delivery of CRISPR/Cas9 system and treatment of HPV infection associated cervical cancer.

Persistent high-risk HPV infection is the main factor for cervical cancer. HPV E7 oncogene plays an important role in HPV carcinogenesis. Down-regulation of E7 oncogene expression could induce growth inhibition in HPV-positive cells and thus treats HPV related cervical cancer. Here we developed a non-virus gene vector based on poly(amide-amine)-poly(ß-amino ester) hyperbranched copolymer (hPPC) for the delivery of CRISPR/Cas9 system to specifically cleave HPV E7 oncogene in HPV-positive cervical cancer cells. The diameter of polyplex nanoparticles (NPs) formed by hPPCs/linear poly(ß-amino ester) (PBAE) and plasmids were approximately 300 nm. These hPPCs/PBAE-green fluorescence protein plasmids polyplex NPs showed high transfection efficiency and low toxicity in cells and mouse organs. By cleaving HPV16 E7 oncogene, reducing the expression of HPV16 E7 protein and increasing intracellular retinoblastoma 1 (RB1) amount, hPPCs/PBAE-CRISPR/Cas9 therapeutic plasmids polyplex NPs, especially highly branched hPPC1-plasmids polyplex NPs, exhibited strong growth inhibition of cervical cancer cells in vitro and xenograft tumors in nude mice. Together, the hPPCs/PBAE polyplex NPs to deliver HPV16 E7 targeted CRISPR/Cas9 system in this study could potentially be applied to treat HPV-related cervical cancer.

An effective vaginal gel to deliver CRISPR/Cas9 system encapsulated in poly (ß-amino ester) nanoparticles for vaginal gene therapy.

BACKGROUND: Gene therapy has held promises for treating specific genetic diseases. However, the key to clinical application depends on effective gene delivery. METHODS: Using a large animal model, we developed two pharmaceutical formulations for gene delivery in the pigs' vagina, which were made up of poly (ß-amino ester) (PBAE)-plasmid polyplex nanoparticles (NPs) based two gel materials, modified montmorillonite (mMMT) and hectorite (HTT). FINDINGS: By conducting flow cytometry of the cervical cells, we found that PBAE-GFP-NPs-mMMT gel was more efficient than PBAE-GFP-NPs-HTT gel in delivering exogenous DNA intravaginally. Next, we designed specific CRISPR/SpCas9 sgRNAs targeting porcine endogenous retroviruses (PERVs) and evaluated the genome editing efficacy in vivo. We discovered that PERV copy number in vaginal epithelium could be significantly reduced by the local delivery of the PBAE-SpCas9/sgRNA NPs-mMMT gel. Comparable genome editing results were also obtained by high-fidelity version of SpCas9, SpCas9-HF1 and eSpCas9, in the mMMT gel. Further, we confirmed that the expression of topically delivered SpCas9 was limited to the vagina/cervix and did not diffuse to nearby organs, which was relatively safe with low toxicity. INTERPRETATION: Our data suggested that the PBAE-NPs mMMT vaginal gel is an effective preparation for local gene therapy, yielding insights into novel therapeutic approaches to sexually transmitted disease in the genital tract. FUNDING: This work was supported by the National Science and Technology Major Project of the Ministry of science and technology of China (No. 2018ZX10301402); the National Natural Science Foundation of China (81761148025, 81871473 and 81402158); Guangzhou Science and Technology Programme (No. 201704020093); National Ten Thousand Plan-Young Top Talents of China, Fundamental Research Funds for the Central Universities (17ykzd15 and 19ykyjs07); Three Big Constructions-Supercomputing Application Cultivation Projects sponsored by National Supercomputer Center In Guangzhou; the National Research FFoundation (NRF) South Africa under BRICS Multilateral Joint Call for Proposals; grant 17-54-80078 from the Russian Foundation for Basic Research.