Salmonellosis remains one of the most prevalent bacterial enteric diseases in swine production worldwide. Tilmicosin (TMS) is widely used in swine bacterial infections, but its oral application is limited by gastric degradation and intense bitterness, reducing bioavailability and therapeutic efficacy. This study aimed to develop and evaluate pilot-scale TMS-loaded nanostructured lipid carriers (TMS@NLCs) to overcome these obstacles. Herein, TMS@NLCs were fabricated at 100-L scale via high-pressure homogenization. Physicochemical properties of TMS@NLCs were characterized, along with stability in simulated gastrointestinal fluids and in vitro antibacterial activity. Palatability was assessed by piglet drinking behavior and electronic tongue. Pharmacokinetics and therapeutic efficacy against Salmonella infection were evaluated in piglets. TMS@NLCs exhibited uniform spherical morphology with mean diameter 231.9 ± 33.3 nm, high encapsulation efficiency (95.50 ± 3.31%) and drug loading (4.703 ± 0.179%). Notably, they also showed excellent gastric stability, and superior taste-masking evidenced by reduced bitterness and 34% higher water intake versus commercial TMS solution. TMS@NLCs enhanced intracellular Salmonella clearance and increased oral bioavailability. In Salmonella-infected piglets, TMS@NLCs significantly reduced bacterial loads, attenuated inflammation and restored intestinal barrier, with no detectable adverse effects. Together, pilot-scale TMS@NLCs successfully overcome the inherent limitations of pronounced bitterness and gastric instability associated with the raw drug. This formulation demonstrates exceptional performance in enhancing oral bioavailability and antibacterial efficacy, highlighting its substantial potential for clinical veterinary translation.

Postoperative distant metastasis and high recurrence rate causes a dilemma in treating triple-negative breast cancer (TNBC) owing to its unforeseeable invasion into various organs or tissues. The wealth of nutrition provided by vascular may facilitate the proliferation and angiogenesis of cancer cells, which further enhance the rates of postoperative metastasis and recurrence. Chemotherapy, as a systemic postoperative adjuvant therapy, is generally applied to diminish recurrence and metastasis of TNBC. Herein, an halofuginone-silver nano thermosensitive hydrogel (HTPM&AgNPs-gel) was prepared via a physical swelling method. The in vitro anticancer efficacy of HTPM&AgNPs-gel was analyzed by investigating cell proliferation, migration, invasion, and angiogenesis capacity. Furthermore, the in vivo anti-cancer activity of HTPM&AgNPs-gel was further appraised through the tumor suppression, anti-metastatic, anti-angiogenic, and anti-inflammatory ability. The optimized HTPM&AgNPs-gel, a thermosensitive hydrogel, showed excellent properties, including syringeability, swelling behavior, and a sustained release effect without hemolysis. In addition, HTPM&AgNPs-gel was confirmed to effectively inhibit the proliferation, migration, invasion, and angiogenesis of MDA-MB-231 cells. An evaluation of the in vivo anti-tumor efficacy demonstrated that HTPM&AgNPs-gel showed a stronger tumor inhibition rate (68.17%) than did HTPM-gel or AgNPs-gel used alone and exhibited outstanding biocompatibility. Notably, HTPM&AgNPs-gel also inhibited lung metastasis induced by residual tumor tissue after surgery and further blocked angiogenesis-related inflammatory responses. Taken together, the suppression of inflammation by interdicting the blood vessels adjoining the tumor and inhibiting angiogenesis is a potential strategy to attenuate the recurrence and metastasis of TNBC. HTPM&AgNPs-gel is a promising anticancer agent for TNBC as a local postoperative treatment.

Canine mammary tumors (CMT) can severely compromise the life quality of the affected dogs through local recurrence, distant metastases and ultimately succumb to death. Recently, more attention has been given to the potential antimetastatic effect of maduramicin (MAD) on breast cancer. However, its poor aqueous solubility and toxicity to normal tissues limit its clinical application. Therefore, to address the drawbacks of MAD and enhance its anticancer and antimetastatic effects, MAD-loaded TPGS polymeric micelles (MAD-TPGS) were prepared by a thin-film hydration technique. The optimized MAD-TPGS exhibited excellent size distribution, stability and improved water solubility. Cellular uptake assays showed that TPGS polymer micelles could enhance drug internalization. Moreover, TPGS synergistically improved the cytotoxicity of MAD by targeting mitochondrial organelles, improving reactive oxygen species levels and reducing the mitochondrial transmembrane potential. More importantly, MAD-TPGS significantly impeded the metastasis of tumor cells. In vivo results further confirmed that, in addition to exhibiting excellent biocompatibility, MAD-TPGS exhibited greater antitumor efficacy than free MAD. Interestingly, MAD-TPGS displayed superior suppression of CMT metastasis via tail vein injection compared to oral administration, indicating its suitability for intravenous delivery. Overall, MAD-TPGS could be applied as a potential antimetastatic cancer agent for CMT.

Contamination by pathogens, such as bacteria, can irritate a wound and prevent its healing, which may affect the physical fitness of the infected person. As such, the development of more novel nano-biomaterials able to cope with the inflammatory reaction to bacterial infection during the wound healing process to accelerate wound healing is required. Herein, a halofuginone‑silver nano thermosensitive hydrogel (HTPM&AgNPs-gel) was prepared via a physical swelling method. HTPM&AgNPs-gel was characterized based on thermogravimetric analysis, differential scanning calorimetry, morphology, injectability, and rheological mechanics that reflected its exemplary nature. Moreover, HTPM&AgNPs-gel was further tested for its ability to facilitate healing of skin fibroblasts and exert antibacterial activity. Finally, HTPM&AgNPs-gel was tested for its capacity to accelerate general wound healing and treat bacterially induced wound damage. HTPM&AgNPs-gel appeared spherical under a transmission electron microscope and showed a grid structure under a scanning electron microscope. Additionally, HTPM&AgNPs-gel demonstrated excellent properties, including injectability, temperature-dependent swelling behavior, low loss at high temperatures, and appropriate rheological properties. Further, HTPM&AgNPs-gel was found to effectively promote healing of skin fibroblasts and inhibit the proliferation of Escherichia coli and Staphylococcus aureus. An evaluation of the wound healing efficacy demonstrated that HTPM&AgNPs-gel had a more pronounced ability to facilitate wound repair and antibacterial effects than HTPM-gel or AgNPs-gel alone, and exhibited ideal biocompatibility. Notably, HTPM&AgNPs-gel also inhibited inflammatory responses in the healing process. HTPM&AgNPs-gel exhibited antibacterial, anti-inflammatory, and scar repair features, which remarkably promoted wound healing. These findings indicated that HTPM&AgNPs-gel holds great clinical potential as a promising and valuable wound healing treatment.

The small-molecule alkaloid halofuginone (HF) is obtained from febrifugine. Recent studies on HF have aroused widespread attention owing to its universal range of noteworthy biological activities and therapeutic functions, which range from parasite infections and fibrosis to autoimmune diseases. In particular, HF is believed to play an excellent anticancer role by suppressing the proliferation, adhesion, metastasis, and invasion of cancers. This review supports the goal of demonstrating various anticancer effects and molecular mechanisms of HF. In the studies covered in this review, the anticancer molecular mechanisms of HF mainly included transforming growth factor-β (TGF-β)/Smad-3/nuclear factor erythroid 2-related factor 2 (Nrf2), serine/threonine kinase proteins (Akt)/mechanistic target of rapamycin complex 1(mTORC1)/wingless/integrated (Wnt)/β-catenin, the exosomal microRNA-31 (miR-31)/histone deacetylase 2 (HDAC2) signaling pathway, and the interaction of the extracellular matrix (ECM) and immune cells. Notably, HF, as a novel type of adenosine triphosphate (ATP)-dependent inhibitor that is often combined with prolyl transfer RNA synthetase (ProRS) and amino acid starvation therapy (AAS) to suppress the formation of ribosome, further exerts a significant effect on the tumor microenvironment (TME). Additionally, the combination of HF with other drugs or therapies obtained universal attention. Our results showed that HF has significant potential for clinical cancer treatment.

Parasitic diseases are the most destructive and pervasive infectious diseases in the world, killing tens of thousands of people annually and causing huge economic losses. Halofuginone is a halogenated derivative of febrifugine isolated and extracted from the plant Changshan, which has strong antiprotozoal activity. Compared with febrifugine, halofuginone has less toxic and side effects, which makes it more advantageous in disease treatment. In recent years, the biological activities of halofuginone in cancer, fibrosis and autoimmune diseases have attracted extensive attention. In human clinical practice, the research on the effects of halofuginone on Duchenne muscular dystrophy and solid tumors has entered the stage of clinical trials. In veterinary clinical practice, halofuginone hydrobromide and halofuginone lactate have been authorized by the FDA and the EU for the prevention and treatment of poultry coccidiosis and ruminant cryptosporidiosis, respectively. Moreover, halofuginone also has efficient inhibition on protozoa parasites such as Plasmodium, Toxoplasma, Theileria, and Leishmania. Aminoacyl-tRNA synthetases are emerging targets for the treatment of parasitic diseases, this review summarizes the effect of halofuginone on various protozoa parasites and the related mechanisms of inhibiting prolyl-tRNA synthetase, and hope to provide a reference for the subsequent theoretical research and clinical application of halofuginone antiprotozoal.

Coccidiosis is a worldwide epidemic intestinal disease with high incidence, which causes huge economic losses. Halofuginone hydrobromide (HF) is widely applied as an effective anticoccidial drug in the poultry industry. However, its therapeutic efficacy is severely restrained due to toxic effects, poor aqueous solubility and low permeability. Nanotechnology can improve the biological effect of drugs, and thus, reduce administered doses and toxic effects. The objective of this study was to investigate the therapeutic and preventive potential of novel HF-loaded D-α-tocopherol polyethylene glycol 1000 succinate (TPGS) polymer micelles (HTPM) for preventing coccidiosis in chickens. The HTPM were approximately spherical with a hydrodynamic diameter of 12.65 ± 0.089 nm, a zeta potential of 8.03 ± 0.242 mV, a drug loading of 14.04 ± 0.12%, and an encapsulation efficiency of 71.1 ± 4.15%. HF was encapsulated in the polymer micelles through interactions with TPGS, as characterized by X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Cellular take up assays showed that TPGS polymer micelles could enhance drug internalization to alleviate intestinal apoptosis induced by coccidiosis and promote the necrosis of second-generation merozoites of E. tenella. Notably, clinical trials proved that 1.5 mg L-1 HTPM had a stronger anticoccidial effect on E. tenella than that of 3 mg kg-1 HF premix. Amplicon sequencing identified that HTPM could alleviate coccidiosis by restoring the structure of the gut microbiome. These findings indicated that the anticoccidial efficacy of HF was significantly enhanced after being encapsulated in polymer micelles, and further demonstrated the potential protective application of nano-encapsulating anticoccidial drugs as a promising approach to control coccidiosis in poultry. In summary, HTPM hold huge potential as an effective therapeutic agent for coccidiosis.

Background Halofuginone (HF)-loaded TPGS polymeric micelles (HTPM) were successfully fabricated using the thin-film hydration technique. HTPM via intravenous injection have been demonstrated to exert an excellent anticancer effect against triple-negative breast cancer (TNBC) cells and subcutaneous xenografts. In the present study, we further explored the potential treatment effect and mechanism of orally administered HTPM alone and in combination with surgical therapy on TNBC in subcutaneous and orthotopic mouse models. Methods Herein, the stability and in vitro release behavior of HTPM were first evaluated in the simulated gastrointestinal fluids. Caco-2 cell monolayers were then used to investigate the absorption and transport patterns of HF with/without encapsulation in TPGS polymeric micelles. Subsequently, the therapeutic effect of orally administered HTPM was checked on subcutaneous xenografts of TNBC in nude mice. Ultimately, orally administered HTPM, combined with surgical therapy, were utilized to treat orthotopic TNBC in nude mice. Results Our data confirmed that HTPM exhibited good stability and sustained release in the simulated gastrointestinal fluids. HF was authenticated to be a substrate of P-glycoprotein (P-gp), and its permeability across Caco-2 cell monolayers was markedly enhanced via heightening intracellular absorption and inhibiting P-gp efflux due to encapsulation in TPGS polymeric micelles. Compared with HF alone, HTPM showed stronger tumor-suppressing effects in subcutaneous xenografts of MDA-MB-231 cells when orally administered. Moreover, compared with HTPM or surgical therapy alone, peroral HTPM combined with partial surgical excision synergistically retarded the growth of orthotopic TNBC. Fundamentally, HTPM orally administered at the therapeutic dose did not cause any pathological injury, while HF alone led to weight loss and jejunal bleeding in the investigated mice. Conclusion Taken together, HTPM could be applied as a potential anticancer agent for TNBC by oral administration.

Background Halofuginone hydrobromide (HF) is a synthetic analogue of the naturally occurring quinazolinone alkaloid febrifugine, which has potential therapeutic effects against breast cancer, however, its poor water solubility greatly limits its pharmaceutical application. D-α-tocopherol polyethylene glycol 1000 succinate (TPGS) is a water-soluble derivative of vitamin E, which can self-assemble to form polymeric micelles (PMs) for encapsulating insoluble anti-tumor drugs, thereby effectively enhancing their anti-cancer effects. Methods HF-loaded TPGS PMs (HTPMs) were manufactured using a thin-film hydration technique, followed by a series of characterizations, including the hydrodynamic diameter (HD), zeta potential (ZP), stability, drug loading (DL), encapsulation efficiency (EE), and in vitro drug release. The anti-cancer effects and potential mechanism of HTPMs were investigated in the breast cell lines MDA-MB-231 and MCF-7, and normal breast epithelial cell line Eph-ev. The breast cancer-bearing BALB/c nude mouse model was successfully established by subcutaneous injection of MDA-MB-231 cells and used to evaluate the in vivo therapeutic effect and safety of the HTPMs. Results The optimized HTPMs had an HD of 17.8±0.5 nm and ZP of 14.40±0.1 mV. These PMs exhibited DL of 12.94 ± 0.46% and EE of 90.6 ± 0.85%, along with excellent storage stability, dilution tolerance and sustained drug release in pH-dependent manner within 24 h compared to free HF. Additionally, the HTPMs had stronger inhibitory effects than free HF and paclitaxel against MDA-MB-231 triple-negative breast cancer cells, and little toxicity in normal breast epithelial Eph-ev cells. The HTPMs induced cell cycle arrest and apoptosis of MDA-MB-231 by disrupting the mitochondrial membrane potential and enhancing reactive oxygen species formation. Evaluation of in vivo anti-tumor efficacy demonstrated that HTPMs exerted a stronger tumor inhibition rate (68.17%) than free HF, and exhibited excellent biocompatibility. Conclusion The findings from this study indicate that HTPMs holds great clinical potential for treating triple-negative breast cancer.

One novel 2-(2-phenylethyl)chromone glycoside was isolated from the EtOH extract of the stems of Aquilaria sinensis. Its structure was elucidated by spectroscopic methods as 6,4′-dimethoxy-3′-hydroxy-2-(2-phenylethyl)chromone 7-O-β-D-glucopyranoside. Compound 1 was assigned the trivial name aquilarinoside C.

Background: Halofuginone hydrobromide (1) is recognized as an effective drug against several species of Eimeria (E.) in poultry. In this paper, we describe a convenient and low cost preparation method for the compound, as well as primary validation of its activity. Methods: First, 7-bromo-6-chloroquinazolin-4(3H)-one (2) was prepared from m-chlorotoluene by a conventional process, and then chloroacetone was creatively introduced in two steps. Finally, halofuginone hydrobromide (1) was obtained from 7-bromo-6-chloro-3-(3-cholroacetonyl) quinazolin-4(3H)-one (4) by a four-step reaction sequence including condensation, cyclization, deprotection and isomerization. The structures of the relative intermediates and target compound were characterized by melting point, IR, MS and 1H-NMR. Besides, the protective effect of compound 1-supplemented chicken diet at doses of 6, 3 and 1.5 mg per 1 kg were evaluated on chickens infected with E. tenella, by reduction in mortality, weight loss, fecal oocyst excretion and gut pathology, respectively. Results: Halofuginone hydrobromide (1) was prepared successfully by and improved and innovative method based on traditional research. Moreover, the synthesized halofuginone hydrobromide significantly exhibited an anti-coccidial property. Conclusions: The fruitful work described in this Communication has resulted in halofuginone hydrobromide, which has a good pharmaceutical development prospects, becoming more available for large-scale production.

[Objectives]The aim of this paper was to make preparation and quality investigation of tilorone dihydrochloride. On the other hand,drug effect on induction of interferon was also primarily proved. [Methods]Tilorone was synthesized by the substitution reaction of 2,7-dihydroxy-9H-fluoren-9-one with 2-diethylaminoethyl chloride hydrochloride,which was then acidified to give tilorone dihydrochloride. The structure of target compound was characterized by Infrared Spectroscopy（ IR）, 1 H Nuclear Magnetic Resonance（ 1 H MNR） and mass spectrum（ MS）. By the method of pharmacopeia,physiochemical properties and content determination were identified in sequence for primary specification establishment. Tilorone dihydrochloride was dissolved at suitable concentration permitting an inoculum of 250 mg·kg -1 （ mice） orally. After tilorone challenged,mice were bled for IFN-α1assay via ELISA method.[Results]Tilorone dihydrochloride was synthesized in good yield of 93% and stable for quality control. In addition,tilorone could characteristically induce an unusually delayed and prolonged interferon response with peak level of 2 000 pg·m L -1 observed from12 h to 24 h. [Conclusions]The compound was efficient to prepare,and was stable and effective to induce obvious interferon responses,which will have broader prospects of development.

Tilorone dihydrochloride (1) has great potential for inducing interferon against pathogenic infection. In this paper, we describe a convenient preparation method for 2,7-dihydroxyfluoren-9-one (2), which is a usual pharmaceutical intermediate for preparing tilorone dihydrochloride (1). In the novel method, methyl esterification of 4,4′-dihydroxy-[1,1′-biphenyl]-2-carboxylic acid (4) was carried out under milder conditions with higher yield and played an important role in the preparation of compound 2. The structures of the relative intermediates and target compound were characterized by melting point, IR, MS, and 1H-NMR. Furthermore, the synthesized tilorone dihydrochloride exhibited an obvious effect on induction of interferon-α (IFN-α) in mice within 12 h, and the peak level was observed until 24 h. This fruitful work has resulted in tilorone dihydrochloride becoming available in large-scale and wide application in clinics, which has a good pharmaceutical development prospects.

Recently, increased reactive oxygen species (ROS) levels and altered redox status in cancer cells have become a novel therapeutic strategy to improve cancer selectivity over normal cells. It has been known that silver nanoparticles (AgNPs) display anti-leukemic activity via ROS overproduction. Hence, we hypothesized that AgNPs could improve therapeutic efficacy of ROS-generating agents against leukemia cells. In the current study, N-(4-hydroxyphenyl)retinamide (4-HPR), a synthetic retinoid, was used as a drug model of ROS induction to investigate its synergistic effect with AgNPs. The data exhibited that AgNPs with uniform size prepared by an electrochemical method could localize in the lysosomes, mitochondria and cytoplasm of SHI-1 cells. More importantly, AgNPs together with 4-HPR could exhibit more cytotoxicity and apoptosis via overproduction of ROS in comparison with that alone. Taken together, these results reveal that AgNPs combined with ROS-generating drugs could potentially enhance therapeutic efficacy against leukemia cells, thereby providing a novel strategy for AgNPs in leukemia therapy.

Solid H2O2 was prepared by anhydrous sodium sulfate,30% H2O2 solution and sodium chloride.Using the single-factor experimental design method,the impact of the material ratio,reaction temperature,reaction time,drying temperature on product yield and H2O2 content was studied to determine the optimum preparation technology.In addition,suspension quantitative germicidal test was carried out to know the germicidal efficacy.The results showed that the product was of a higher H2O2 content and good quality in the following condition,25 mL 30% H2O2,adding 32 g NaSO4,4 g NaCl,with the reaction time of 30 minutes,reaction temperature of 40 ℃,and the drying temperature of 60 ℃.10.8 g·L-1 H2O2 thimerosal killed 99.9% in average of Escherichia coli,Staphylococcus aureus with a 10 min contact and 100% in average of Bacillus cereus with a 1 h contact.Conclusion:solid H2O2 prepared in this test has a good bactericidal effect,and the preparation process is simple.

建立用高效液相色谱法(HPLC)测定地昔尼尔含量的方法。采用Hypersil NH2色谱柱(5μm,250 mm×4.6 mm),流动相:乙腈∶水(0.1%甲酸)为40∶60,紫外检测波长为270 nm,进样量20μL,流速为0.8 mL/m in,柱温30℃。结果显示,地昔尼尔在0.78～100μg/mL范围内,其峰面积与浓度的线性关系良好,回归方程为:A=329 232 C-44 761,R2=0.999 9。日内、日间精密度RSD均小于2%,稳定性试验RSD为0.61%,重复性试验RSD为1.45%,回收率测定结果为100.72%。本法操作简单,结果准确,分析快速,适用于地昔尼尔的含量测定。

The inhibitory effects of fluoroquinolones on the enzyme activity, protein levels and mRNA expression of liver cytochrome P450 (CYP) 1A and 3A were investigated in male broiler chicks. Enrofloxacin (20 mg/kg), sarafloxacin (8 mg/kg) and marbofloxacin (5.5 mg/kg) were administrated in drinking water for 7 consecutive days. A cocktail of the probe drugs caffeine and dapsone was used to determine CYP1A and 3A activity. Western blot analysis and real-time PCR were used to determine the effects on protein levels of CYP1A and 3A, and on CYP1A4, 1A5, 3A37 mRNA levels. Enrofloxacin increased the half-life of elimination for both caffeine and dapsone, and decreased expression of CYP1A and 3A protein. Marbofloxacin decreased the metabolism of caffeine and expression of CYP1A protein. However, no change in mRNA expression was observed for any treatment group. This suggested that high doses of enrofloxacin and marbofloxacin, but not sarafloxacin, inhibit CYP in chick liver raising the possibility of drug–drug interaction when using these compounds.

This experiment was conducted to study the pharmacokinetics of 1florfenicol sulfonate in broiler chickens plasma.Twelve broiler chickens were used to give single intramuscular injection of florfenicol sulfonate at a dosage of 20 mg/kg.The concentrations of florfenicol sulfonate in plasma were determined by HPLC.The pharmacokinetics parameters were calculated by 3P97 computer program and fitted a one-compartment open model with weight by 1/C.The main parameters of florfenicol sulfonate in broiler chickens were as follows:T1/2 kα:(0.28±0.04)h,T1/2 Ke:(2.06±0.06)h,Cmax:(4.17±0.12)μg/mL,Tmax:(0.92±0.09)h,AUC:(16.89±0.35)μg/mL,V/F(c):(3.52±0.13)L/kg,CL/F(s):(1.19±0.03)L/(kg·h),Ke:(0.34±0.01)/h,kα:(2.57±0.37)/h,A:(6.56±0.38)μg/mL.The results showed that the distribution of florfenicol sulfonate was rapid and wide in the broiler chickens,the concentration of plasma was high and elimination was quick.