Recent advances in biological detection with rolling circle amplification: design strategy, biosensing mechanism, and practical applications.

Rolling circle amplification (RCA) is a simple and isothermal DNA amplification technique that is used to generate thousands of repeating DNA sequences using circular templates under the catalysis of DNA polymerase. Compared to alternating temperature nucleic acid amplification such as polymerase chain reaction (PCR) amplification, RCA is more suitable for on-spot detection without the need for an expensive thermal cycler. In this study, the principle and classification of RCA are introduced, and the applications of RCA in the detection of pathogenic bacteria, nucleic acid tumor markers, viruses, and proteins are reviewed. Finally, the perspectives of RCA in biological detection are anticipated. The RCA method has a great potential for biological detection. This review aims to provide references for the further development and application of the RCA technique in biosensors.

Anlotinib enhances the antitumor activity of radiofrequency ablation on lung squamous cell carcinoma.

Anlotinib is a novel molecular targeted drug that has been approved for the treatment of lung adenocarcinoma. Currently these agents are rarely used in the treatment of lung squamous cell carcinoma (LSCC). Bronchoscope-guided radiofrequency ablation (RFA) is a new strategy proposed for the treatment of LSCC that is able to alleviate the obstruction of the respiratory tract caused by LSCC by direct destruction of the tumor tissues. The presence work aims to reveal whether Anlotinib could enhance the antitumor activity of RFA on LSCC cells. The results from real-time PCR (qPCR) confirmed overexpression of targets of anlotinib activity, including receptor tyrosine kinase or the MPAK/PI3K-AKT pathway kinases, in LSCC tissues. Treatment with anlotinib inhibited the survival, in vitro invasion, and migration of LSCC cells. Moreover, the antitumor effects of RFA were investigated using a rodent model of LSCC. The combination of RFA and anlotinib treatment enhanced the antitumor effect of RFA treatment. We propose a combinative strategy of RFA and anlotinib as a novel approach for successful management of LSCC.

ZafA Gene Is Important for Trichophyton mentagrophytes Growth and Pathogenicity.

Trichophyton mentagrophytes is a common fungal pathogen that causes human and animal dermatophytosis. Previous studies have shown that zinc deficiency inhibits T. mentagrophytes growth, and the ZafA gene of T. mentagrophytes can code the functionally similar zinc finger transcriptional factor that can promote zinc ion absorption; however, the impact of ZafA on virulence and pathogenicity remains undetermined. To assess its gene function, the ZafA mutant, ZafA-hph, and the ZafA complemented strain, ZafA+bar, were constructed via Agrobacterium tumefaciens-mediated transformation. Polymerase chain reaction and Southern blot analyses were used to confirm the disruption. In vitro growth capacity and virulence analyses comparing ZafA-hph with wild-type T. mentagrophytes and ZafA+bar showed that ZafA-hph's growth performance, reproduction ability, and zinc ion absorption capacity were significantly lower than the wild-type T. mentagrophytes and ZafA+bar. ZafA-hph also showed weak hair biodegradation ability and animal pathogenicity. Thus, the significant decrease in T. mentagrophytes' growth ability and virulence was due to a lack of the zinc-responsive activity factor rather than the transformation process. This study confirmed that the T. mentagrophytes' zinc-responsive activity factor plays important roles in the pathogen's growth, reproduction, zinc ion absorption, and virulence. This factor is important and significant for effectively preventing and controlling T. mentagrophytes infections.

Transcriptome sequencing and analysis of zinc-uptake-related genes in Trichophyton mentagrophytes.

BACKGROUND: Trichophyton mentagrophytes is an important zoonotic dermatophytic (ringworm) pathogen; causing severe skin infection in humans and other animals worldwide. Fortunately, commonly used fungal skin disease prevention and treatment measures are relatively simple. However, T. mentagrophytes is primarily studied at the epidemiology and drug efficacy research levels, yet current study has been unable to meet the needs of clinical medicine. Zinc is a crucial trace element for the growth and reproduction of fungi and other microorganisms. The metal ions coordinate within a variety of proteins to form zinc finger proteins, which perform many vital biological functions. Zinc transport regulatory networks have not been resolved in T. mentagrophytes. The T. mentagrophytes transcriptome will allow us to discover new genes, particularly those genes involved in zinc uptake. RESULT: We found T. mentagrophytes growth to be restricted by zinc deficiency; natural T. mentagrophytes growth requires zinc ions. T. Mentagrophytes must acquire zinc ions for growth and development. The transcriptome of T. mentagrophytes was sequenced by using Illumina HiSeq™ 2000 technology and the de novo assembly of the transcriptome was performed by using the Trinity method, and functional annotation was analyzed. We got 10,751 unigenes. The growth of T. mentagrophytes is severely inhibited and there were many genes showing significant up regulation and down regulation respectively in T. mentagrophytes when zinc deficiency. Zinc deficiency can affect the expression of multiple genes of T. mentagrophytes. The effect of the zinc deficiency could be recovered in the normal medium. And we finally found the zinc-responsive activating factor (ZafA) and speculated that 4 unigenes are zinc transporters. We knocked ZafA gene by ATMT transformation in T. mentagrophytes, the result showed that ZafA gene is very important for the growth and the generation of conidia in T. mentagrophytes. The expression of 4 zinc transporter genes is potentially regulated by the zinc-responsive activating factor. The data of this study is also sufficient to be used as a support to study T. mentagrophytes. CONCLUSION: We reported the first large transcriptome study carried out in T. mentagrophytes where we have compared physiological and transcriptional responses to zinc deficiency, and analyzed the expression of genes involved in zinc uptake. The study also produced high-resolution digital profiles of global genes expression relating to T. mentagrophytes growth.

Synthesis of gold Nanoshells through Improved Seed-Mediated Growth Approach: Brust-like, in Situ Seed Formation.

Gold nanoshells have shown great potentials in various fields. However, the widely used seed-mediated growth method based on a silica template for gold nanoshells is a complex and time-consuming procedure. In this work, mercaptosilica was first used as a template to synthesize gold nanoshells through improved seed-mediated growth method. It is verified that gold seeds were formed and attached onto the mercaptosilica nanospheres through Brust-like, in situ process, which makes this method extremely time-saving and easy to manipulate. Importantly, the key factors affecting the in situ process were demonstrated, allowing fine control on the synthesis in a highly reproducible manner. The as-synthesized nanoshells are monodisperse with well-defined morphology and tunable near-IR plasmon resonance. Furthermore, other metal nanoparticles such as Pt and Pd could be grafted onto the surface of mercaptosilica nanospheres through the same Brust-like, in situ process. These provide new insights into seed attachment, and the improved seed-mediated growth approach based on Brust-like, in situ seed formation will take an important step forward toward the widespread application of gold nanoshells.

Controlled synthesis of multilayered gold nanoshells for enhanced photothermal therapy and SERS detection.

It can be streamlined: A facile and controllable approach for the fabrication of core/shell-structured multilayer gold nanoshells with uniform nanosize, monodispersity, and tunable plasmonic properties has been successfully developed by utilizing an organosilica layer as the dielectric spacer layer.

Dual-responsive polypseudorotaxanes based on block-selected inclusion between polyethylene-block-poly(ethylene glycol) diblock copolymers and 1,4-diethoxypillar[5]arene.

Based on the selective recognition of the polyethylene (PE) block of polyethylene-block-poly(ethylene glycol) (PE-b-PEG) by 1,4-diethoxypillar[5]arene (DEP5A), two novel thermo and competitive guest (1,4-dibromobutane or hexanedinitrile) responsive polypseudorotaxanes (PPRs) have been successfully constructed. The formation of PPRs both in solution and in the solid state was demonstrated by (1)H NMR, 2D NOESY, and WAXD analyses. TGA data illustrate that PPRs exhibit higher thermal stability than their precursor diblock copolymers. Moreover, intriguing porous disk-like aggregates are produced by electrospraying of PPRs in CHCl3 and the self-assembled structures of PPRs are totally changed by the addition of 1,4-dibromobutane or hexanedinitrile, demonstrating their competitive guest stimuli-responsiveness.

Anti-aggregation colorimetric sensing of cysteine using silver nanoparticles in the presence of Pb2.

Using silver nitrate as the silver source and sodium borohydride as the reducing agent, we synthesized negatively charged silver nanoparticles (AgNPs). Subsequently, the AgNPs solution was mixed with positively charged lead ions, resulting in AgNPs aggregation via electrostatic interactions. This led to a color change in the solution from yellow to purple and eventually to blue-green. Our study focused on a colorimetric method that exhibited high selectivity and sensitivity in detecting cysteine using AgNPs-Pb2+ as a sensing probe. Upon the introduction of cysteine to the AgNPs-Pb2+ system, the absorbance of AgNPs increased at 396 nm and decreased at 520 nm. The formation of a complex between cysteine and lead ions prevented the aggregation of silver nanoparticles, enabling the colorimetric detection of cysteine. The relationship between the concentration of ΔA396/A520 and cysteine showed linearity within the range of 0.01 to 0.1 µM; the regression equation of the calibration curve is ΔA396/A520 = 9.0005c - 0.0557 (c: µM), with an R2 value of 0.9997. The detection limit was found to be 3.8 nM (S/N = 3). This method demonstrated exceptional selectivity and sensitivity for cysteine and was effectively used for the determination of cysteine in urine. Our findings offer a new perspective for the future advancement of anti-aggregation silver nanocolorimetry.

Nitrogen-doped carbon material NCM-T heterogeneously catalyzed liquid-phase hydrogenation of nitrobenzene to aniline.

As an important chemical intermediate, aniline is primarily produced industrially through catalytic hydrogenation of nitrobenzene. Herein, a series of nitrogen-doped carbon materials (referred to as NCM-T, with T denoting the roasting temperature (°C)) were prepared through high-temperature roasting of sucrose and melamine for the heterogeneous catalytic liquid-phase hydrogenation of nitrobenzene to aniline. A preliminary study of the involved reaction mechanism was performed by combining the results of material characterisation and catalyst evaluation. Experimental results showed that the graphitic N content and the defective sites simultaneously affected the performance of NCM-T in catalysing the hydrazine hydrate reduction in the nitrobenzene hydrogenation reaction. The catalyst NCM-800 was reacted in an ethanol solution with hydrazine hydrate as the reducing agent at 80 °C for 5 h. Notably, the nitrobenzene conversion rate was up to 94%, and the aniline selectivity was 100%. The turnover frequency (TOF) could reach up to 7.9 mol g-1 h-1, and after five recycling cycles, only a small loss of catalytic activity was observed. This shows that the prepared catalyst is a recyclable catalyst that can be used for reducing the nitrobenzene from hydrazine hydrate to aniline.

Analyzing the characteristics of respiratory microbiota after the placement of an airway stent for malignant central airway obstruction.

Malignant central airway stenosis is treated with airway stent placement, but post-placement microbial characteristics remain unclear. We studied microbial features in 60 patients post-stent placement, focusing on changes during granulation tissue proliferation. Samples were collected before stent (N = 29), after stent on day 3 (N = 20), and after granulation tissue formation (AS-GTF, N = 43). Metagenomic sequencing showed significant respiratory tract microbiota changes with granulation tissue. The microbiota composition, dominated by Actinobacteria, Firmicutes, and Proteobacteria, was similar among the groups. At the species level, the AS-GTF group exhibited significant differences, with Peptostreptococcus stomatis and Achromobacter xylosoxidans enriched. Analysis based on tracheoesophageal fistula presence identified Tannerella forsythia and Stenotrophomonas maltophilia as the main differential species, enriched in the fistula subgroup. Viral and fungal detection showed Human gammaherpesvirus 4 and Candida albicans as the main species, respectively. These findings highlight microbiota changes after stent placement, potentially associated with granulation tissue proliferation, informing stent placement therapy and anti-infective treatment optimization. IMPORTANCE: Malignant central airway stenosis is a life-threatening condition that can be effectively treated with airway stent placement. However, despite its clinical importance, the microbial characteristics of the respiratory tract following stent insertion remain poorly understood. This study addresses this gap by investigating the microbial features in patients with malignant central airway stenosis after stent placement, with a specific focus on microbial changes during granulation tissue proliferation. The findings reveal significant alterations in the diversity and structure of the respiratory tract microbiota following the placement of malignant central airway stents. Notably, certain bacterial species, including Peptostreptococcus stomatis and Achromobacter xylosoxidans, exhibit distinct patterns in the after-stent granulation tissue formation group. Additionally, the presence of tracheoesophageal fistula further influences the microbial composition. These insights provide valuable references for optimizing stent placement therapy and enhancing clinical anti-infective strategies.

Source profile study of VOCs unorganized emissions from typical aromatic devices in petrochemical industry.

Volatile organic compounds (VOCs) are significant pollutants generated during the processes of petroleum refining and chemical production. Aromatic hydrocarbons, in particular, pose a great risk to human health. Nevertheless, unorganized emissions of VOCs from typical aromatics units remain poorly studied and reported. Therefore, it is vital to achieve precise control over aromatic hydrocarbons while managing VOCs. In this study, two typical aromatics production devices in petrochemical enterprises, namely aromatics extraction devices and ethylbenzene devices, were selected. The fugitive emissions of VOCs from the process pipelines in the units were investigated. Samples were collected and transferred using the EPA bag sampling method and HJ 644 and analyzed using gas chromatography-mass spectrometry. The results indicated that a total of 112 VOCs were emitted during the six rounds of sampling in the two types of devices, with alkanes (61 %), aromatic hydrocarbons (24 %), and olefins (8 %) being the primary types of VOCs emitted. The results also revealed the unorganized emissions characteristic substances of VOCs in the two types of devices, with slight differences in the types of VOCs emitted. The study found significant differences in the detection concentrations of aromatic hydrocarbons and olefins, as well as the types of detected chlorinated organic compounds (CVOCs), between the two sets of aromatics extraction units in distinct regions. These differences were closely related to the processes and leakages in the devices and can be effectively controlled by enhancing leak detection and repair (LDAR) and other measures. This article offers guidance for compiling VOCs emission inventories and improving the management of VOCs emissions in petrochemical enterprises by refining the source spectrum at the device scale. The findings are significant for analyzing VOCs unorganized emission factors and promoting safe production in enterprises.

Constructed a self-powered sensing platform based on nitrogen-doped hollow carbon nanospheres for ultra-sensitive detection and real-time tracking of double markers.

Acute myocardial infarction (AMI) is acute necrosis of a portion of the myocardium caused by myocardial ischemia, which seriously threatens people's health and life safety. Its early diagnosis is a difficult problem in clinical medicine. Research has found that the abnormal expression of microRNA-199a (miR-199a) and microRNA-499 (miR-499) was closely related to AMI disease. In this work, we took advantage of the structural advantages of nitrogen-doped hollow carbon nanospheres (N-HCNSs) to design an ultra-sensitive, portable real-time monitoring visual self-powered biosensor system, which based on dual-target miRNAs triggered catalytic hairpin assembly (CHA) for sensitive detection of miR-199a and miR-499. In addition, the capacitor and the smartphone are introduced into the system to realize the secondary improvement of system sensitivity and portable real-time visual monitoring. Under optimized conditions, in the linear range of 0.1-100000 aM, the detection limits of miR-199a and miR-499 are 0.031 and 0.027 aM, respectively. At the same time, the ultra-sensitive detection of miRNAs is realized in the serum sample, and the recovery rate of miR-199a and miR-499 are 98.0-106.0% (RSD: 0.6-8.1%) and 94.0-109.7% (RSD: 1.8-7.7%), respectively. The method is simple, sensitive and can be used for real-time tracking and portable monitoring of related diseases.

A three-dimensional interconnected molybdenum disulfide/multi-walled carbon nanotubes cathode with enlarged interlayer spacing for aqueous zinc-ion storage.

Layered molybdenum disulfide (MoS2) shows tremendous prospect as cathode material for aqueous zinc-ion batteries (AZIBs) due to the two-dimensional zinc ions (Zn2+) diffusion channels and tunable interlayer spacing. However, it is subjected to sluggish insertion/extraction kinetics, inferior electronic conductivity and inadequate active capacities. Herein, a three-dimensional (3D) interconnected MoS2/multi-walled carbon nanotubes (MWCNTs) framework is proposed to address these issues. Importantly, the MWCNTs cores offer interconnection routes for fast electrons and zinc ions transport, the expanded spacing of MoS2 interlayer with 1.05 nm can facilitate rapid Zn2+ intercalation/extraction, and the confined MoS2 layers in inner MWCNTs can mitigate the agglomeration and restacking of MoS2 nanosheets. Benefitting from the confined structural configuration, sufficient active surface and 3D structural stability, the MoS2/MWCNTs as AZIBs cathode delivers a large initial reversible capacity of 218.3 mAh/g and high coulombic efficiency of 78.2 % at 0.1 A/g. Additionally, the 3D interconnected cathode maintains nearly intact structure after a fierce galvanostatic charge/discharge process, resulting in large retained capacities of 126.3 mAh/g at 1 A/g after 650 cycles and 101.1 mAh/g at 3 A/g after 1000 cycles. This work offers a novel strategy for the structure design of two-dimensional materials to develop high-performance cathodes for AZIBs.

The self-powered electrochemical biosensing platform with multi-amplification strategy for ultrasensitive detection of microRNA-155.

A self-powered biosensor (SPB) was constructed for the ultra-sensitive detection of microRNA-155 (miR-155) by combining a capacitor/enzymatic biofuel cell (EBFC), a strategy of rolling circle amplification (RCA) and a digital multimeter (DMM). The experimental results show that the sensitivity of the assembled EBFC-SPB can reach 15.85 µA/pM with the action of matching capacitor, which is 513% of that without capacitor (3.09 µA/pM). This achieves the first signal amplification. Furthermore, when the target miR-155 triggers RCA, electrons are continuous generated and flow to the biocathode through the external circuit to catalyze the reduction of oxygen and release [Ru(NH3)6]3+ electron acceptor. This achieves the second signal amplification. Finally, DMM is used to convert the signal into instantaneous current and amplify it for real-time reading. This achieves the third signal amplification. Therefore, the limit of detection (LOD) of the developed biosensor is as low as 0.17 fM (S/N = 3), and the linear range is between 0.5 fM and 10,000 fM, indicating that the EBFC-SPB has a broad application prospect for cancer marker of miR-155 with ultrasensitive detection.

Novel senescence associated gene, YPEL3, is repressed by estrogen in ER+ mammary tumor cells and required for tamoxifen-induced cellular senescence.

Estrogen signaling plays an important role in breast carcinogenesis. An increased understanding of estrogen gene targets and their effects will allow for more directed and effective therapies for individuals with breast cancer, particularly those with estrogen receptor positive tumors resistant to tamoxifen therapy. Here, we identify YPEL3 as a growth suppressive protein downregulated by estrogen in estrogen receptor positive breast cancer cell lines. Estrogen repression of YPEL3 expression was found to be independent of p53 but dependent on estrogen receptor alpha expression. Importantly, YPEL3 expression, which is induced by the removal of estrogen or treatment with tamoxifen triggers cellular senescence in MCF-7 cells while loss of YPEL3 increases the growth rate of MCF-7 cells. Taken together these findings suggest that YPEL3 may represent a potential target for directed hormonal therapy for estrogen receptor positive breast cancer patients.

Detection of Ag+ ions via an anti-aggregation mechanism using unmodified gold nanoparticles in the presence of thiamazole.

The present study proposed a novel and highly selective and sensitive method for Ag+ ion detection based on gold nanoparticles (AuNPs) anti-aggregation. Thiamazole can induce AuNPs aggregation due to electrostatic interactions, which result in color transitions in the AuNPs solution from red to blue. However, the presence of Ag+ ions results in the preferential combination of the pyridinic nitrogen of thiamazole with the Ag+ ions. In addition, the Ag+ ions oxidize the sulfhydryl groups(-SH), which inhibit AuNPs aggregation and prompt a color change from blue to red. As a result, the present study established a method for Ag+ ion determination by AuNPs-thiamazole colorimetric probe based on the aforementioned anti-aggregation mechanism. The probe dynamic range was easily tuned via adjustments of the thiamazole amount. The relationship between the Ag+ concentration and AuNPs aggregation was monitored by ultraviolet-visible light (UV-Vis) spectroscopy at a dynamic range of 0.1 nM-9 µM and at a detection limit of 0.042 nM. The river water and tap water recovery analysis validated the successful operation of this colorimetric sensor in environmental monitoring.

Isolation and characterization of endothelial progenitor cells from canine bone marrow.

Endothelial progenitor cells (EPC) are located predominantly in the bone marrow. These cells are useful for treating human vascular diseases; they also are a possible target for restricting blood vessel growth for tumors. Little is known about canine EPC. We investigated a bone marrow EPC isolation method that combines the whole bone marrow culture method and the differential adherent speed method using stillborn canines. MTT proliferation, flow cytometry detection, Dil-ac-LDL uptake, FITC-UEA-1 binding and matrigel assays were used to identify and characterize EPC. We isolated two types of EPC: early EPC and late EPC. We found that isolated cells produced typical colony and cobblestone morphology, and were positive for CD31, CD34, CD133 and VEGFR-2. Significant differences were observed in the intensity of expression between early and late EPC, which suggests their different roles during angiogenesis and vasculogenesis. Both early and late EPC were positive for Dil-ac-LDL and FITC-UEA-1, and displayed tube formation when re-suspended in matrigel, both of which are important functional criteria for identifying EPC. Our method is a novel, effective and efficient way to produce enriched EPC.

Interconnected MoS2 on 2D Graphdiyne for Reversible Sodium Storage.

In this study, graphdiyne (GDY) was first reported as a substrate material for sodium-ion batteries (SIBs). The creative hybridization of GDY and molybdenum disulfide (MoS2) endows the composite with unique heterostructural and morphological advantages that boost the charge transport rate and enhance the battery discharge properties. Electrochemical results indicated that the MoS2@GDY anode displays a considerable discharge capacity of up to 328 mAh g-1 at 1000 mA g-1. A capacity retention of 93% even at testing current back to 200 mA g-1 suggests superior rate characteristics. An outstanding stable cyclic performance of 217 mAh g-1 is obtained at a high testing density. The attractive results not only demonstrate that GDY could be used not only as an effective conductive substrate to prevent the host material from agglomerating in the electrochemical process but also provide a novel design for fabricating efficient electrode materials for future energy-storage systems.

Clinical effects of cisplatin plus recombinant human endostatin (rh-endostatin) intratumoral injection on malignant central airway obstruction: a retrospective analysis of 319 cases.

BACKGROUND: Primary lung cancer with severe central airway obstruction (CAO) is often life-threatening. In this study, we investigated the clinical efficacy and safety of cisplatin plus recombinant human endostatin (rh-endostatin) intratumoral injection in treatment of malignant central airway obstruction (MCAO) caused by primary squamous cell lung cancer. METHODS: We retrospectively analyzed patients with MCAO caused by primary squamous cell lung cancer treated with and without bronchoscopic intratumoral injection of cisplatin plus rh-endostatin between January 2007 and June 2016. RESULTS: A total of 206 patients received cisplatin plus rh-endostatin intratumoral injection, and 113 without injection. Dyspnea grade, degree of stenosis, quality of life and lung function of all patients were significantly improved at 1 week after treatment compared with baseline. Both groups achieved good airway patency (97.1% vs. 93.8%, P=0.156). Followed up at 2 months, all parameters were improved in the injection group compared with baseline, while no statistical differences were observed in the non-injection group (P>0.05). The injection group achieved airway patency in 155 (75.2%) of 206 patients, which was significantly superior to the non-injection group [20 (17.7%) of 113, P<0.001]. In addition, the restenosis rate of the injection group was lower compared with the non-injection group (22.5% vs. 81.1%, P<0.001, respectively). No serious complications were observed in two groups. CONCLUSIONS: Cisplatin plus rh-endostatin intratumoral injection is effective and safe for the therapy of MCAO caused by primary squamous cell lung cancer.

Testosterone propionate can promote effects of acellular nerve allograft-seeded bone marrow mesenchymal stem cells on repairing canine sciatic nerve.

Peripheral human nerves fail to regenerate across long tube implants (>2 cm), and tissue-engineered nerve grafts represent a promising treatment alternative. The present study aims to investigate the testosterone propionate (TP) repair effect of acellular nerve allograft (ANA) seeded with allogeneic bone marrow mesenchymal stem cells (BMSCs) on 3-cm canine sciatic nerve defect. ANA cellularized with allogeneic BMSCs was implanted to the defect, and TP was injected into the lateral crus of the defected leg. The normal group, the autograft group, the ANA + BMSCs group, the ANA group, and the nongrafted group were used as control. Five months postoperatively, dogs in the TP + ANA + BMSCs group were capable of load bearing, normal walking, and skipping, the autograft group and the ANA + BMSCs group demonstrated nearly the same despite a slight limp. The compound muscle action potentials (CMAPs) on the injured side to the uninjured site in the TP + ANA + BMSCs group were significantly higher than that in the ANA + BMSCs group [CMAPs ratio at A: F(3, 20) = 191.40; 0.02, CMAPs ratio at B: F(3, 20) = 43.27; 0.01]. Masson trichrome staining revealed that in the TP + ANA + BMSCs group, both the diameter ratio of the myelinated nerve and the thickness ratio of regenerated myelin sheath were significantly larger than that in the other groups [the diameter of myelinated nerve fibers: F(3, 56) = 13.45; P < .01, the thickness ratio of regenerated myelin sheath: F(3, 56) = 51.25; P < .01]. In conclusion, TP could significantly increase the repairing effects of the ANA + BMSCs group, and their combination was able to repair 3-cm canine sciatic nerve defect. It therefore represents a promising therapeutic approach.