The pathogenesis of hepatocellular carcinoma (HCC) emphasizes metabolic disorders. HCC patients showed abnormally low expression of Acyl-CoA dehydrogenase short chain (ACADS). This study aimed to elucidate the clinical significance and mechanistic role of ACADS in HCC. We investigated the expression patterns and significance of ACADS in HCC by analyzing multiple public databases and clinical samples (Chip data). Immunohistochemistry was employed to observe the expression levels of ACADS in HCC tissues. In vitro experiments involved silencing or overexpressing ACADS in HCC cell lines, with protein expression levels determined by Western blotting. Functional validation included CCK-8, Transwell, and scratch wound healing assays. TOPFlash and FOPFlash reporter gene assays, co-immunoprecipitation, and immunofluorescence were used to explore the interaction between ACADS and β-catenin. ACADS was low expressed in HCC and was clinically associated with vascular invasion, TNM stage, and AFP levels. The low ACADS expression in HCC patients was negatively correlated with their survival. Overexpression of ACADS significantly suppressed the viability, migration, and invasive capacity of HCC cells, whereas silencing ACADS had the opposite effect. Mechanistically, co-immunoprecipitation experiments indicated that there was an interaction between ACADS and β-catenin. Overexpression of ACADS inhibited β-catenin activity and resulted in decreased nuclear β-catenin translocation and increased its cytoplasmic level. Immunofluorescence results also showed a decrease in β-catenin nuclear import following ACADS overexpression, whereas silencing ACADS led to an enhancement of its nuclear translocation. ACADS emerges as a potentially valuable biomarker for HCC prognosis, exhibiting tumor-suppressive functions in HCC by participating in the regulation of β-catenin activity.

To investigate the factors influencing the healing of open fractures in circumpolar latitude region seawater immersion conditions. A femoral fracture model was established in ninety 6-to-8-week-old male Sprague-Dawley rats, randomly assigned to five groups (n = 18 per group): (1) fracture only, (2) fracture with circumpolar seawater immersion, (3) fracture with low-temperature isotonic solution immersion, (4) fracture with aseptic circumpolar seawater immersion, and (5) fracture with low-temperature aseptic circumpolar seawater immersion. Fractures were confirmed postoperatively by radiographs on days 7, 21, and 42. Micro-CT and H E staining were performed on day 42 to assess bone healing. Bacterial cultures from internal fixation devices were analyzed on day 3. Blood samples were collected on days 3, 7, and 14 to assess leukocyte and neutrophil counts, and serum ALP and VEGF levels were measured on days 7, 14, and 21. Pathogenic microorganisms in the seawater were identified by metagenomic analysis. Fracture healing and callus formation rates were compared using the Log-rank test. X-ray, micro-CT, and histological analyses revealed significantly impaired fracture healing in the group exposed to circumpolar seawater immersion compared to the fracture-only group (P < 0.05). Bacterial colony counts on internal fixation devices were highest in the circumpolar seawater group (P < 0.05). Leukocyte and neutrophil levels were significantly elevated in this group on days 3 and 7 (P < 0.05), with no significant differences observed on day 14 (P > 0.05). Serum ALP and VEGF levels were significantly reduced on days 7, 14, and 21 (P < 0.05), although ALP levels on day 21 showed no significant difference (P > 0.05). Log-rank analysis indicated that the bone union and callus maturation rates were significantly lower in the circumpolar seawater group compared to the other four groups. Metagenomic analysis identified Flavobacterium, Rhodobacter, and Bacteroides as the dominant pathogens in circumpolar seawater. This study demonstrates that hyperosmolarity, low temperature, and exposure to opportunistic pathogens under circumpolar seawater conditions collectively delay open fracture healing. Among these factors, opportunistic pathogens exert the most significant impact, highlighting microbial contamination as the primary barrier to bone regeneration in such environments and providing direction for future therapeutic strategies. not applicable.

Objective: The aim of this study was to evaluate the efficacy, safety, and surgical outcomes of 2-stage management, namely preoperative endoscopic retrograde cholangiopancreatography (ERCP) + laparoscopic cholecystectomy (ERCP+LC) or LC + postoperative ERCP (LC+ERCP), as well as 1-stage management, LC + laparoscopic common bile duct exploration (LCBDE) for treating patients with gallstones and common bile duct stones (CBDS). Methods: This retrospective study analyzed the data of 180 patients with common bile duct stones (CBDS) who were admitted to the Department of General Surgery at Tongji Hospital, Tongji University, between January 2019 and June 2021. The study included 3 groups: ERCP+LC (group 1), LC+ERCP (group 2), and LC+LCBDE (group 3), each consisting of 60 patients. Clinical metrics of the patients were collected and compared among the groups. Results: Group 3 had the shortest operation duration and hospital stay compared with group 1 and group 2. In addition, group 3 had the lowest long-term postoperative complications, particularly the recurrence rate of CBDS. The total cost was also the lowest in group 3. Furthermore, patients in group 3 had the lowest postoperative amylase levels. All patients in the study achieved successful stone clearance. There were no significant differences in the conversion to other procedures rate, postoperative alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, and mortality among the three groups (P > 0.05). Conclusions: Both 1-stage management and 2-stage management are effective treatments for CBDS. The LC+LCBDE management is a safe treatment option, offering shorter hospital stays and operation duration, lower costs, and fewer complications.

目的 探讨肝胆外科肿瘤患者发生利奈唑胺相关性血小板减少的危险因素,为患者临床安全用药提供依据.方法 选取本院 2017年 1月至 2021年 12月间应用利奈唑胺进行抗感染治疗的肿瘤患者,根据给予利奈唑胺后是否出现血小板减少,将患者分为血小板减少组和未发生组.比较两组患者一般资料与实验室指标,采用多因素logistic回归分析筛选发生利奈唑胺相关性血小板减少的危险因素.结果 研究共纳入 104例患者,其中接受外科手术患者 84例,未接受外科手术患者 20例.利奈唑胺相关性血小板减少发生率为 24.0%.两组患者性别、年龄、利奈唑胺使用时长、血小板计数、白细胞计数、谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素、肌酐、估算肾小球滤过率有显著性差异(P<0.05);logistic回归分析结果提示,年龄≥60岁(OR=7.093;P=0.017)、利奈唑胺使用时长≥12 d(OR=4.399;P=0.035)、基础血小板计数≤200×109/L(OR=8.470;P=0.004)、基础AST≥50 U/L(OR=15.465;P<0.001)、基础白细胞计数≥11×109/L(OR=11.436;P=0.001)是肿瘤患者发生利奈唑胺相关性血小板减少的危险因素.结论 给肝胆外科肿瘤患者应用利奈唑胺时,医师需关注患者是否发生血小板减少的不良反应,尤其是年老、长疗程、基础血小板低、基础肝功能差及基础白细胞计数高的患者.

Background The transaxillary approach of breast augmentation is the most popular method in Asia, but longer period of recovery was observed in spite of the assistance of endoscope. Objectives Introducing the ultrasonic dissection devices might be a solution to minimizing tisue damage thus relieving pain and shortening the period of recovery. Method Between March 2020 and September 2022, we retrospectively reviewed the cases of 122 patients underwent endoscopic augmentation mammoplasty via the transaxillary approach using either the monopolar electrotome (ME) alone or assisted with Harmonic Scalpel (HS) in defining the retropectoral pocket and severing the pectoralis major muscle. Result The total drainage volume was significantly lower in the HS group than ME group (74.33 ± 48.81 vs. 180.30 ± 125.10 mL; p < 0.0001). VAS score of the first 24 hour after surgery of the ME group was significantly higher than that of the HS group (6.10 ± 1.27 vs. 2.88 ± 1.29, p < 0.0001). Operation time in HS group was reduced compared to ME group (113.1 ± 14.46 mins vs. 131.3 ± 35.51 mins, p < 0.001). The duration of drainage placement (1.08 ± 0.27 vs. 2.72 ± 1.18 days) and hospital stay after surgery (3.08 ± 0.42 vs. 5.64 ± 2.78 days; p < 0.0001) were largely reduced in HS group. Conclusion The assistance of Harmonic Scalpel significantly reduced total postoperative drainage, relieved pain and shortened operation time, length of drainage placement and hospital stay compared to using monopolar electrotome alone in endoscopic-assisted transaxillary dual-plane augmentation mammaplasty. Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Introduction: Lacking of normal innervation increases the chance of chronic wounds and recurrence of ulceration. Various rodent models are designed to reveal nerve-wound relationship but present many limitations to mimic human wound which heals primarily by re-epithelialization rather than contraction in rodents. This article tested a modified rat model of denervated wound healing to better mimic clinical common denervated wounds. Material and Methods: The wounds formed on right hind paws of 18 SD rats served as the experimental (denervated) group and the left side as contra-lateral control (non-denervated). The denervation was achieved through sciatic and femoral nerve co-transection and the control side underwent sham-surgery 3 days prior to a skin punch wound formation on both sides. Wound closure rate was calculated under digital photographing. Loss of innervation and affected healing process was confirmed by histological analyses. Results: Truncation of the sciatic and femur nerve successfully denervated the skin of the hind paw and resulted in a significantly declined healing rate, prolonged inflammation, weakened dermal contraction, hindered macrophage recruitment, retarded re-epithelialization and collagen deposition, decreased angiogenesis and epidermal proliferation, and persisted epidermal apoptosis compared to the innervated contra-lateral control. Conclusion: Wound on denervated dorsal pedis in rats can be used to study denervated skin healing in multiple histological process. We believe that this model will assist in understanding the underlying mechanism of nerve-wound relationship and identifying new treatment strategies that can be more rapidly translated into clinical practice.

ObjectiveWe aim to clarify the vascular and nerve anatomy of the breast and combine it with an ultrasound knife to use in transaxillary endoscopic biplane breast augmentation.MethodsThis study is a retrospective review of patients undergoing transaxillary endoscopic biplane breast augmentation between October and October 2021. Related variables were collected using a standardized data collection template. The detailed process of the transaxillary endoscopic biplane breast augmentation under anatomy instruction is carefully described in this study, and the postoperative effect was closely observed.ResultsSixty-three female patients underwent transaxillary endoscopic biplane breast augmentation. The average implants volume counted 242.46 ± 31.34 cc, and the average operation time was 155.92 ± 22.34 min. Patients were followed up for a mean of 13.67 months (range, 3–27 months), and most of the patients achieved good postoperative results and no severe complications and were satisfied with both appearance and function.ConclusionsThe application of anatomy combined with an ultrasound knife in transaxillary endoscopic biplane breast augmentation is a promising way to achieve good breast shapes with high patient satisfaction and is worthy of clinical promotion and application.

Age‐related changes to the face pertinent anatomy and important aspects of autologous fat grafting have been widely applied in facial rejuvenation. Various types of autologous fat products (Macrofat, Microfat, SEFF, nanofat, and SVF‐gel) with different properties and applicabilities have been introduced and available for surgeons.

Diabetes mellitus (DM) is a metabolic disease, now prevalent worldwide, which is characterized by a relative or absolute lack of insulin secretion leading to chronically increased blood glucose levels. Diabetic patients are often accompanied by multiple macrovascular complications, such as coronary heart disease, hypertension, macrovascular arteriosclerosis, and microvascular complications. Microvascular complications include diabetic kidney injury, diabetic encephalopathy, and diabetic foot, which reduce the quality of life and survival status of patients. Mesenchymal stem cell exosomes (MSC-Exos) possess repair functions similar to MSCs, low immunogenicity, and ease of storage and transport. MSC-Exos have been proven to possess excellent repair effects in repairing various organ damages. This study reviews the application of MSC-Exos in the treatment of DM and its common complications. MSC-Exos may be used as an effective treatment for DM and its complications.

Background Xanthelasma palpebrarum is a type of human xanthoma that occurs on the skin of human eyelids and is a benign skin lesion. Pingyangmycin (also known as bleomycin A5) is one of the 13 components of bleomycin. The aim of this study was to explore the efficacy of intralesional bleomycin and pingyangmycin in the treatment of xanthoma based on histopathological observations in animal experimental research. Methods An animal model of xanthoma was established by feeding rabbits with a high-cholesterol diet. Pingyangmycin and bleomycin interfered with the skin xanthoma of the animal model. Skin tissue specimens were stained with hematoxylin-eosin and oil red O to evaluate the effect of the intervention. Results A xanthoma animal model was established. Pingyangmycin and bleomycin could reduce the abnormal lipid deposition in the lesion area of the skin xanthoma of the animal, via a local injection. In addition, pingyangmycin was more effective than bleomycin in eliminating lipid deposition in rabbit skin xanthoma.

The hypoxia-inducible factor 1-alpha (HIF-1a) pathway plays a key role in regulating angiogenesis during wound healing. However, the diabetic condition hampers the stabilization of HIF-1a and thus inhibits the subsequent angiogenesis, and meanwhile, the function and phenotype transition of macrophage are impaired in the diabetic condition, which leads to prolonged and chronic inflammation. Both angiogenesis inhibition and inflammatory dysfunction make diabetic wound healing a major clinical challenge. Here, borosilicate (BS), a new group of bioceramics with a coupled network of interconnected [BO3] and [SiO4] which can incorporate therapeutic ions such as Cu2+, is synthesized and combined with silk fibroin (SF), a biocompatible natural amino acid polymer whose composition and structure are similar to a natural extracellular matrix (ECM), to obtain a compound system which can transform into a SF-MA-BS hydrogel under UV radiation via methacryloyloxy (MA) groups modified on both BS and SF. When in use, the compound system can thoroughly spread to the whole wound surface and be in situ photo-cross-linked to form an integral SF-MA-BS hydrogel that firmly adheres to the wound, protects the wound from external contamination, and further spontaneously promotes wound regeneration by releasing therapeutic ions. The wound repair of Streptozotocin-induced diabetic rats shows that diabetic wound healing is obviously accelerated by SF-MA-BS, interestingly the HIF-1a pathway is restored via interaction between HIF-1a and Cu2+, and angiogenesis is therefore enhanced. Meanwhile, inflammation is well regulated by SF-MA-BS, and long-term detrimental inflammation is avoided. These findings indicate that the SF-MA-BS hydrogel regenerates diabetic wounds, and further clinical trials are anticipated.

Excessive scar formation has adverse physiological and psychological effects on patients; therefore, a therapeutic strategy for rapid wound healing and reduced scar formation is urgently needed. Herein, bilayered thiolated alginate/PEG diacrylate (BSSPD) hydrogels were fabricated for sequential release of small extracellular vesicles (sEVs), which acted in different wound healing phases, to achieve rapid and scarless wound healing. The sEVs secreted by bone marrow derived mesenchymal stem cells (B-sEVs) were released from the lower layer of the hydrogels to promote angiogenesis and collagen deposition by accelerating fibroblast and endothelial cell proliferation and migration during the early inflammation and proliferation phases, while sEVs secreted by miR-29b-3p-enriched bone marrow derived mesenchymal stem cells were released from the upper layer of the hydrogels and suppressed excessive capillary proliferation and collagen deposition during the late proliferation and maturation phases. In a full-thickness skin defect model of rats and rabbit ears, the wound repair rate, angiogenesis, and collagen deposition were evaluated at different time points after treatment with BSSPD loaded with B-sEVs. Interestingly, during the end of the maturation phase in the in vivo model, tissues in the groups treated with BSSPD loaded with sEVs for sequential release (SR-sEVs@BSSPD) exhibited a more uniform vascular structure distribution, more regular collagen arrangement, and lower volume of hyperplastic scar tissue than tissues in the other groups. Hence, SR-sEVs@BSSPD based on skin repair phases was successfully designed and has considerable potential as a cell-free therapy for scarless wound healing.

Diced cartilage is a significant alternative approach to cartilage grafting. However, the viability and biomechanical properties of diced cartilage grafts remain to be improved, and the role of perichondrium is largely neglected. This study aimed to evaluate the histological and biomechanical effects of perichondrium on custom-shaped diced cartilage grafts constructed via a high-density porous polyethylene mold. Seven New Zealand rabbits were used. Unilateral auricular cartilage was harvested and divided into 2 parts, with or without perichondrium, diced into 1 × 1 × 0.5 mm cubical pieces, and filled into high-density porous polyethylene molds. Three grafts with the perichondrium removed and 3 with the perichondrium preserved were implanted subcutaneously at the dorsum. The grafts underwent biomechanical and histological tests 4, 8, and 12 weeks after the implantation. The diced cartilage merged into integrated blocks without observable resorption in both groups at each time point. Additionally, the retention rate of weight was higher in the perichondrium-preserved group (P < 0.05). We observed regenerated cartilage that stained positively for type II collagen and glial fibrillary acidic protein (GFAP). A greater area of regenerated cartilage and higher scores of GFAP staining were observed in the perichondrium-preserved group (P < 0.05). The yield stress and modulus of elasticity were also higher in the perichondrium-preserved grafts from week 8 after implantation (P < 0.05). Diced cartilage grafts with a custom shape can be constructed using a high-density porous polyethylene mold. The preservation of perichondrium can improve graft viability and biomechanical properties. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266.

Background Xanthelasma palpebrarum (XP) is the most common type of cutaneous xanthoma and has been treated with intralesional injection of pingyangmycin effectively. However, bleomycin, which has the same effect in antitumor activity as pingyangmycin, has not been applied in the treatment of XP. Aims To explore and assess the treatment of xanthelasma by intralesional injection of bleomycin, which has been widely used as an antitumor antibiotic, for the replacement of pingyangmycin. Methods Intralesional injection of different concentrations of bleomycin was administered to 44 xanthelasma lesions of 24 patients who have never been treated before, divided into two groups according to age. Photographs were taken and analyzed to assess the therapeutic efficiency. Patients were then followed up for 6-24 months. Results All the lesions resolved after 1 month of treatment with the intralesional injection of different concentrations of bleomycin. There was no significant difference observed between the two groups. No severe complications had occurred. Conclusion The treatment of XP with intralesional injection of bleomycin is minimally invasive, safe, and effective. Consequently, it also has good cosmetic outcome with no adverse effects.

Background XP is a flat xanthoma that usually presents as bilateral, symmetrical, soft, yellowish papules over the eyelids. The etiology of XP is unknown, but it may be related to complex heterozygous mutations. Aims To investigate the lipid profiles, important clinical characteristics, and low-density lipoprotein receptor (LDLR) gene mutation in the patients suffering from xanthelasma palpebrarum (XP) with nonfamilial hypercholesterolemia. Patients/Methods The prospective study included 25 individuals of XP with nonfamilial hypercholesterolemia, and 30 controls neither with XP nor familial hypercholesterolemia (FH). Each one underwent detailed clinical examination, serum lipid profile, and LDLR gene detection at the 400th exon and the 1246th exon. Results In our study, patients with XP were often not present with FH and family history. The mean serum cholesterol (CHOL) (5.20 +/- 1.82) and the prevalence of carotid atherosclerosis (4.34 +/- 0.78) were significantly higher in study group, while there was no statistically significant difference between two groups in terms of triglyceride (TG) (P = .38) and low-density lipoprotein (LDL) (P = .23). Unusually, the mean levels of high-density lipoprotein (HDL) (1.59 +/- 0.31) in the study group were much higher than the controls (1.31 +/- 0.30), and the LDLR gene mutation was not found. Conclusion Clinical and serum lipid profiles indicated that XP was sporadic and underlying lipid abnormalities especially higher HDL. XP did not occur with mutations in the LDLR gene at the two exons. We suggested too high HDL level may be contributed to pathogenesis of XP and XP is often not associated with FH.

Microglia are the primary immune cells in the central nervous system with functional plasticity. They can be activated into M1 and M2 phenotypes when neuroinflammation-related diseases occur. M1 phenotype cells produce pro-inflammatory mediators that cause neuroinflammation and the M2 phenotype can secrete anti-inflammatory cytokines that protect neurons from damage. Therefore, inhibiting the M1 phenotype while stimulating the M2 phenotype has been suggested as a potential therapeutic approach for treating neuroinflammation-related diseases. Puerarin has been demonstrated to exert anti-inflammatory and neuroprotective effects. However, the role of puerarin in regulating microglia polarization and its reaction mechanism has not been fully elucidated. In this paper, a metabolomics approach with ultra-performance liquid chromatography-mass spectrometry was performed to investigate the metabolic changes of BV-2 cells in different phenotypes and test the effects of puerarin on polarization. Thirty-nine metabolites were identified as the biomarkers related to the polarization of BV-2 cells and puerarin intervention reverted the content of most of the biomarkers. Our study demonstrated that puerarin could play a key role in M1/M2 polarization of BV-2 cells from a perspective of metabolomics, and it could regulate the balance between promotion and suppression of inflammation.

Ebracteolatain A is a phloroglucinol derivative from the root of Euphorbia ebracteolata Hayata, a Traditional Chinese Medicine also known as Langdu. It has been shown to have good inhibitory effects in breast cancer cells. In this study, a simple, rapid, sensitive, and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to study the pharmacokinetics (PKs) and tissue distribution of Ebracteolatain A in rats. Ebracteolatain A and Magnolol (internal standard) were extracted by the simple protein precipitation extraction technique using methanol as the precipitating solvent. Chromatographic separation was performed using the Agilent Poroshell 120 EC-C-18 column with a mobile phase of acetonitrile:0.1% formic acid (70:30, v/v). The protonated analyte was quantitated in negative ionization by MS/MS via multiple reaction monitoring mode. The assay exhibited a linear dynamic range of 2-2000 ng/mL for Ebracteolatain A in biological samples. The lower limit of quantitation was 2 ng/mL. Non-compartmental PK parameters indicated that Ebracteolatain A was well absorbed into the systemic circulation. The absolute bioavailability of Ebracteolatain A was greater when administered by intraperitoneal administration than by oral administration. The tissue distribution study showed that Ebracteolatain A was distributed in the heart, liver, spleen, lung, kidney, brain, stomach, intestine, uterus, ovary, and breast after intravenous injection. The results of this study further our understanding of the in vivo anti-cancer activity of Ebracteolatain A, and shed light on pharmacological strategies that may be useful for the development of novel breast cancer therapeutics. (C) 2019 Elsevier B.V. All rights reserved.

Objective To present our experience and techniques with the use of autologous costal cartilage grafts in Asian rhinoplasty,and to report the surgical results and complications in 86 consecutive rhinoplasty cases.Methods All operations were performed by the first author (Liu AT) with open approach,costal cartilages and perichondrium were used to reconstruct the nasal tip projection according to the tripod theory in rhinoplasty,after removing the previous injection material,L-shaped implant or hypertrophic scar tissue in the tip.Medical charts and operative records were reviewed retrospectively to summary the complications.Nasal dorsum augmentation was done by costal cartilage or I-shaped allograft,sometimes with anterior sheath of rectus abdominis.Patients' subjective satisfaction of the postoperative nasal appearance was self-evaluated with grading (1 worse,2 no change,3 improved,and 4 much improved).Results From September 2015 to March 2017,86 patients underwent rhinoplasty at our hospital.The postoperative follow-up duration was 6 to 20 months.Overall,functional and aesthetic outcome was satisfactory in most patients,and the mean score by the patients' self-evaluation was 3.3 ± 0.6.Graft exposure,mobility,or significant resorption,pneumothorax or significant donor-site pain were not observed.Conclusions Even with minimal complications and morbidities,autologous costal cartilage grafts in Asian rhinoplasty is a versatile and reliable graft material for nasal tip surgery in severe short or saddle nose,contracted nose due to previous L-shaped augmentation and revision rhinoplasty in which the septal cartilage has already been harvested.

Stellera chamaejasme L. has been used as a traditional Chinese medicine for the treatment of scabies, tinea, stubborn skin ulcers, chronic tracheitis, cancer and tuberculosis. A sensitive and selective ultra-high liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of five flavonoids (stelleranol, chamaechromone, neochamaejasmin A, chamaejasmine and isochamaejasmin) of S. chamaejasme L. in rat plasma. Chromatographic separation was accomplished on an Agilent Poroshell 120 EC-C18 column (2.1 × 100 mm, 2.7 μm) with gradient elution at a flow rate of 0.4 mL/min and the total analysis time was 7 min. The analytes were detected using multiple reaction monitoring in positive ionization mode. The samples were prepared by liquid-liquid extraction with ethyl acetate. The UPLC-MS/MS method was validated for specificity, linearity, sensitivity, accuracy and precision, recovery, matrix effect and stability. The validated method exhibited good linearity (r ≥ 0.9956), and the lower limits of quantification ranged from 0.51 to 0.64 ng/mL for five flavonoids. The intra- and inter-day precision were both <10.2%, and the accuracy ranged from -11.79 to 9.21%. This method was successfully applied to a pharmacokinetic study of five flavonoids in rats after oral administration of ethyl acetate extract of S. chamaejasme L.

Cataracts are the most common cause of blindness worldwide affecting tens of millions of people. Here, we report a simple, rapid, sensitive and specific method by ultra performance liquid chromatography-tandem mass spectrometry with the electrospray ionization mode (UPLC-ESI-MS/MS) for quantitation of lanosterol, a possible effective drug for cataracts, in the vitreous humor of rabbits after ocular administration. The injected lanosterol was prepared by dispersing lanosterol molecules into the poly-(DL-lactic acid-co-glycolic acid)-poly(ethylene glycol) -poly-(DL-lactic acid-co-glycolic acid) (PLGA-PEG-PLGA) thermogel solution. The analyte and internal standard (IS, panaxadiol) were extracted by the simple protein precipitation with methanol. The chromatographic separation used an Agilent RRHD SB-Cis column with a methanol mobile phase containing 50 mM of ammonium acetate aqueous solution (with 0.1% formic acid) (95:5, viv). The protonated analyte was quantitated in positive ionization by multiple reaction monitoring (MRM) with a mass spectrometer. The mass transitions iniz 443.5 -> 235 and rn/z 461 -> 127 were used to measure the analyte and IS, respectively. The assay exhibited a linear dynamic range of 1-1250 ngmL(-1) for lanosterol in vitreous samples. The lower limit of quantitation (LLOQ) was 1 ngmL(-1) with a relative standard deviation (RSD) of less than 15%. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 5 min per sample offered a throughput of more than 200 samples per day. This validated method was used to analyze vitreous samples of New Zealand white rabbits for pharmacokinetic studies. The results provided useful information on pharmacological action mechanism of lanosterol and were meaningful for cataract treatment among the elderly population. (C) 2017 Elsevier B.V. All rights reserved.