Enhancing the revelation of key genes and interaction networks in non-small cell lung cancer with major depressive disorder: A bioinformatics analysis.

Background and Aims: Lung cancer is ranked as the second most prevalent form of cancer worldwide. Nonsmall cell lung cancer (NSCLC) represents the predominant histological subtype. Research suggests that one-third of lung cancer patients also experiencing depression. Antidepressants play an indispensable role in the management of NSCLC. Despite significant advancements in treatment, lung cancer patients still face a high mortality rate. Major depressive disorder (MDD) and related antidepressants involved in treatment efficacy and prognosis of NSCLC. However, there has been a lack of screening and analysis regarding genes and networks associated with both NSCLC and MDD. Methods: To investigate the correlation between MDD and NSCLC, our discovery and validation analysis included four datasets from the Gene Expression Omnibus database from NSCLC or MDD. Differential gene expression (DEGs) analysis, GO and KEGG Pathway, and protein-protein interaction network analyzes to identify hub genes, networks, and associated observations link between MDD and NSCLC. Results: The analysis of two datasets yielded a total of 84 downregulated and 52 upregulated DEGs. Pathway enrichment analyzes indicated that co-upregulated genes were enriched in the regulation of positive regulation of cellular development, collagen-containing extracellular matrix (ECM), cytokine binding, and axon guidance. We identified 20 key genes, which were further analyzed using the MCODE plugin to identify two core subnetworks. The integration of functionally similar genes provided valuable insights into the potential involvement of these hub genes in diverse biological processes including angiogenesis humoral immune response regulation inflammatory response organization ECM network. Conclusion: We have identified a total of 136 DEGs that participate in multiple biological signaling pathways. A total of 20 hub genes have demonstrated robust associations, potentially indicating novel diagnostic and therapeutic targets for both diseases.

Per- and polyfluoroalkyl substances in Chinese commercially available red swamp crayfish (Procambarus clarkii): Implications for human exposure and health risk assessment.

The extensive utilization of per- and polyfluoroalkyl substances (PFASs) has led to their pervasive presence in the environment, resulting in contamination of aquatic products. Prolonged exposure to PFASs has been linked to direct hepatic and renal damage, along with the induction of oxidative stress, contributing to a spectrum of chronic ailments. Despite the recent surge in popularity of red swamp crayfish as a culinary delicacy in China, studies addressing PFASs' exposure and associated health risks from their consumption remain scarce. To address this gap, our study investigated the PFASs' content in 85 paired edible tissue samples sourced from the five primary red swamp crayfish breeding provinces in China. The health risks associated with dietary exposure were also assessed. Our findings revealed widespread detection of PFASs in crayfish samples, with short-chain perfluoroalkyl carboxylic acids (PFCAs) exhibiting the highest concentrations. Notably, the total PFAS concentration in the hepatopancreas (median: 160 ng/g) significantly exceeded that in muscle tissue (5.95 ng/g), as did the concentration of every single substance. The hazard quotient of perfluorohexanesulfonic acid (PFHxS) via consuming crayfish during peak season exceeded 1. In this case, a potential total non-cancer health risk of PFASs, which is mainly from the hepatopancreas and associated with PFHxS, is also observed (hazard index>1). Thus, it is recommended to avoid consuming the hepatopancreas of red swamp crayfish. Greater attention should be paid to governance technology innovation and regulatory measure strengthening for short-chain PFASs.

Parabens and triclosan in red swamp crayfish (Procambarus clarkii) from China: Concentrations, tissue distribution and related human dietary intake risk.

Parabens (PBs) and triclosan (TCS) are commonly found in pharmaceuticals and personal care products (PPCPs). As a result, they have been extensively found in the environment, particularly in aquaculture operations. Red swamp crayfish (Procambarus clarkii) consumption has significantly risen in China. Nevertheless, the levels of PBs and TCS in this species and the associated risk to human dietary intake remain undisclosed. This study assessed the amounts of five PBs, i.e., methyl-paraben (MeP), ethyl-paraben (EtP), propyl-paraben (PrP), butyl-paraben (BuP) and benzyl-paraben (BzP), as well as TCS in crayfish taken from five provinces of the middle-lower Yangtze River. MeP, PrP and TCS showed the highest detection rates (hepatopancreas: 46-86 %; muscle: 63-77 %) since they are commonly used in PPCPs. Significantly higher levels of ∑5PBs (median: 3.69 ng/g) and TCS (median: 7.27 ng/g) were significantly found in the hepatopancreas compared to the muscle (median: 0.39 ng/g for ∑5PBs and 0.16 ng/g for TCS) (p < 0.05), indicating bioaccumulation of these chemicals in the hepatopancreas. The estimated daily intake values of ∑5PBs and TCS calculated from the median concentrations of crayfish were 6.44-7.94 ng/kg bw/day and 11.4-14.0 ng/kg bw/day, respectively. Although no health risk was predicted from consuming crayfish (HQ <1), consumption of the hepatopancreas is not recommended.

CENPW knockdown inhibits progression of bladder cancer through inducing cell cycle arrest and apoptosis.

Purpose: The objective of this study was to examine the expression and role of Centromere protein W (CENPW) in bladder cancer (BLCA), as well as its potential mechanistic impact on the progression of BLCA. Methods: In this study, we conducted a comparative analysis of the mRNA expression level of CENPW in BLCA tissues and adjacent normal tissues using data from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Additionally, we investigated the association between CENPW expression and patient prognosis. Furthermore, we performed in vitro and in vivo experiments to assess the impact of CENPW knockdown on various tumor biological phenotypes in BLCA. Finally, we conducted an analysis to elucidate the underlying mechanisms responsible for the observed phenotypic alterations in BLCA. Results: The expression of CENPW was found to be upregulated in BLCA, and its higher expression was associated with a poorer disease-specific survival (DSS). CENPW was found to have close associations with the cell cycle, mitosis, and DNA replication. In vitro and in vivo experiments demonstrated that the inhibition of CENPW led to a suppression of BLCA progression. Specifically, the knockdown of CENPW resulted in cell cycle arrest phase and induced apoptosis in BLCA by potentially inactivating the signal transducer and activator of transcription3 (STAT3) signaling pathway. Conclusion: CENPW has the potential to function as a molecular marker indicating an unfavorable prognosis in BLCA. Additionally, CENPW exhibits promise as a novel therapeutic target for BLCA.

Exploring the correlation of glycolysis-related chondroitin polymerizing factor (CHPF) with clinical characteristics, immune infiltration, and cuproptosis in bladder cancer.

Bladder cancer (BLCA) is a common malignant neoplasm of the urinary system. Glycolysis is an essential metabolic pathway regulated by various genes with implications for tumor progression and immune escape. Scoring the glycolysis for each sample in the TCGA-BLCA dataset was done using the ssGSEA algorithm for quantification. The results showed that the score in BLCA tissues was markedly greater than those in adjacent tissues. Additionally, the score was found to be correlated with metastasis and high pathological stage. Functional enrichment analyses of the glycolysis-related genes showed they were related to roles associated with tumor metastasis, glucose metabolism, cuproptosis, and tumor immunotherapy in BLCA. Using 3 different machine learning algorithms, we identified chondroitin polymerizing factor (CHPF) as a central glycolytic gene with high expression in BLCA. In addition, we showed CHPF is a valuable diagnostic marker of BLCA with an area under the ROC (AUC) of 0.81. Sequencing BLCA 5637 cells after siRNA-mediated CHPF silencing and bioinformatics revealed that CHPF positively correlated with the markers of epithelial-to-mesenchymal transformation (EMT), glycometabolism-related enzymes, and immune cell infiltration. In addition, CHPF silencing inhibited the infiltration of multiple immune cells in BLCA. Genes that promote cuproptosis negatively correlated with CHPF expression and were up-regulated after CHPF silencing. High CHPF expression was a risk factor for overall and progression-free survival of patients who received immunotherapy for BLCA. Finally, using immunohistochemistry, we demonstrated that the CHPF protein had high expression in BLCA, increasing in high-grade tumors and those with muscle invasion. The CHPF expression levels were also positively associated with 18F-fluorodeoxyglucose uptake in PET/CT images. We conclude that the glycolysis-related gene CHPF is an effective diagnostic and treatment target for BLCA.

Psychological distress influences lung cancer: Advances and perspectives on the immune system and immunotherapy.

Lung cancer has the highest incidence rate and mortality worldwide. Moreover, multiple factors may cause heterogeneity in the efficacy of immunotherapy for lung cancer, and preclinical studies have gradually uncovered the promotive effects of psychological distress (PD) on tumor hallmarks. Therefore, treatment targeted at PD may be a vital factor in adjusting and improving immunotherapy for lung cancer. Here, by focusing on the central nervous system, as well as stress-related crucial neurotransmitters and hormones, we highlight the effects of PD on the lung immune system, the lung tumor microenvironment (TME) and immunotherapy, which brings a practicable means and psychosocial perspective to lung cancer treatment.

Four new species of the subgenus Homoneura from Yintiaoling Nature Reserve, China (Diptera: Lauxaniidae: Homoneura).

The subgenus Homoneura Malloch, 1927 is the most diverse in China, with more than 220 known species. Four new species of Homoneura (Homoneura) henanensis group, that is, Homoneura (Homoneura) apiconcava sp. nov., H. (H.) chongqingensis sp. nov., H. (H.) wuxica sp. nov., and H. (H.) yintiaolingica sp. nov., were described from Yintiaoling Nature Reserve, Chongqing, China. A key to the known species of this species group is presented.

Effect and molecular mechanism research of Astragalus membranaceus on inhibiting intestinal absorption of six alkaloids of Aconitum carmichaelii in spleen deficiency rats.

Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

[Co-amorphous technology to improve dissolution and physical stability of silybin].

The present study explored the effect of co-amorphous technology in improving the dissolution rate and stability of silybin based on the puerarin-silybin co-amorphous system prepared by the spray-drying method. Solid-state characterization was carried out by powder X-ray diffraction(PXRD), polarizing microscopy(PLM), Fourier transform infrared spectroscopy(FT-IR), differential scanning calorimetry(DSC), etc. Saturated powder dissolution, intrinsic dissolution rate, moisture absorption, and stability were further investigated. The results showed that puerarin and silybin formed a co-amorphous system at a single glass transition temperature which was higher than that of any crude drug. The intrinsic dissolution rate and supersaturated powder dissolution of silybin in the co-amorphous system were higher than those of the crude drug and amorphous system. The co-amorphous system kept stable for as long as three months under the condition of 40 ℃, 75% relative humidity, which was longer than that of the single amorphous silybin. Therefore, the co-amorphous technology could significantly improve the dissolution and stability of silybin.

Four new species of the genus Luzonomyza Malloch (Diptera, Lauxaniidae) from China.

Four species of the genus Luzonomyza Malloch, 1929 from southwest China are described as new to science: Luzonomyzavittifacies Li & Yang, sp. nov., L.serrata Li & Yang, sp. nov., L.honghensis Li & Yang, sp. nov., and L.brevis Li & Yang, sp. nov. A key to Luzonomyza species is also presented.

Downregulation of lncRNA IGF2-AS-encoded peptide induces trophoblast - cycle arrest.

RESEARCH QUESTION: lncRNA IGF2-AS may be related to early pregnancy loss. Does lncRNA IGF2-AS affect trophoblast cell growth? The aim of the present study was to verify that lncRNA IGF2-AS encodes a polypeptide, IGF2-AS-168aa, and to study its role in the pathogenesis of trophoblasts. DESIGN: A small interfering RNA targeted to the IGF2-AS gene (si-IGF2-AS) was designed and transfected into JEG-3 and JAR cells for in-vitro gene silencing. Quantitative polymerase chain reaction and western blotting were used to determine lncRNA IGF2-AS levels in experimental cells. After IGF2-AS suppression, MTT assay was used to assess cell proliferation and apoptosis was determined by flow cytometry. Target gRNA IGF2-AS-gRNA was designed for knockout conducted the corresponding mRNA. HEK293T cells were transfected with the identified positive clone vectors. Finally, IGF2-AS-168aa was analysed by western blotting after the protein-coding region of the IGF2-AS gene was knocked out by CRISPR/Cas9 gene-editing technology. RESULTS: lncRNA IGF2-AS and IGF2-AS-168aa were significantly downregulated in JEG-3 and JAR cells transfected with si-IGF2-AS (lncRNA IGF2-AS: JAR: NC versus small interfering RNA (siRNA)-1: P = 0.019 NC versus siRNA-2: P = 0.013; JEG-3: NC versus siRNA-1: P = 0.001 NC versus siRNA-2: P = 0.004) (IGF2-AS-168aa: JAR: NC versus siRNA-1: P = 0.030 NC versus siRNA-2: P = 0.018; JEG-3: NC versus siRNA-1: P = 0.004 NC versus siRNA-2: P = 0.001). IGF2-AS gene was incapable of encoding IGF2-AS-168aa after the coding region was successfully knocked out in HEK293T cells. Flow cytometry and the MTT assay revealed that IGF2-AS gene silencing led to cell cycle block in the G1 phase, markedly decreasing cell proliferation and increasing apoptosis. CONCLUSION: The IGF2-AS gene encoded a peptide with a potential function in trophoblast cell cycle arrest.

Improvement of the bioavailability of curcumin by a supersaturatable self nanoemulsifying drug delivery system with incorporation of a hydrophilic polymer: in vitro and in vivo characterisation.

OBJECTIVES: The current study was focused on preparing curcumin (CUR) supersaturated self-nano-emulsion (PI-CUR-SNEDDS) using hydrophilic polymer and to study the influence of polymer precipitation inhibitor on the physicochemical and biopharmaceutical properties of the PI-CUR-SNEDDS. METHODS: PI-CUR-SNEDDS were prepared using hydrophilic polymer in order to maintain the supersaturation of CUR in nano-emulsion solution, artificial gastrointestinal fluid (AGF), and the precipitates formed, and characterised by in vitro dispersion tests, in vitro intestinal absorption and in vivo pharmacokinetic and compared with CUR-SNEDDS. KEY FINDINGS: PI-CUR-SNEDDS prepared with 2% hydroxypropyl methylcellulose 55-60 (HPMC55-60) as precipitation inhibitor (PI) significantly improved the viscosity, physical stability and CUR's equilibrium solubility of nanoemulsion. HPMC55-60 and CUR interact in AGF through intermolecular interactions, form hydrogen bonds, and produce amorphous precipitates. Compared with CUR-SNEDDS, the proportion of CUR in the hydrophilic phase increased by about 3-fold, and apparent permeability coefficient (Papp) in duodenum, jejunum, ileum, and colon increased by 2.30, 3.65, 1.54 and 2.08-fold, respectively, and the area under the plasma concentration-time curve0-12h of PI-CUR-SNEDDS also increased by 3.50-fold. CONCLUSIONS: Our results suggested that HPMC55-60 maintained the CUR supersaturation state by forming hydrogen bonds with CUR, increasing the solution's viscosity and drug solubilisation, thus improving the absorption and bioavailability of CUR.

Release Characteristics of an Essential Oil Component Encapsulated with Cyclodextrin Shell Matrices.

BACKGROUND: Essential oils are poor aqueous solubility and high volatility compounds. The encapsulation of essential oils with Cyclodextrins (CDs) can protect them from adverse environmental conditions and improve their stability. Therefore, increasing the functional capabilities of essential oils when they were used as additives in pharmaceutical and food systems. Additionally, the release of active compounds is an important issue. However, there were few studies about the effect of different CDs on the release of drugs after encapsulation. Therefore, the information on the study of release models is considerably limited. OBJECTIVE: This study aimed to (i) characterize the physico-chemical properties and release behavior of myrcene encapsulated in the four different shell matrices of α-CD, ß-CD, γ-CD and 2-hydroxypropyl-ß- cyclodextrin (HP-ß-CD), which were selected from the perspective of stability, and (ii) determine the release mechanism of myrcene in Inclusion Complexes (ICs). METHODS: ICs of myrcene and four CDs were prepared by freeze-drying. The physico-chemical properties of ICs were fully characterized by laser diffraction particle size analyzer, Scanning Electron Microscope (SEM), Fourier-Transform Infrared spectroscopy (FT-IR) and Differential Scanning Calorimeter (DSC). The release behaviors of ICs at 50, 60, 70 and 80 °C were determined and described by zeroorder or first-order kinetics with the Henderson-Pabis, Peppas, Avrami and Page mathematical models. Moreover, the possible binding modes of ICs were identified with molecular modelling technique. RESULTS: Firstly, the structure of Particle Size Distribution (PSD), FT-IR, DSC and SEM showed that (i) CDs could effectively encapsulate the myrcene molecules, and (ii) the release kinetics were well simulated by Avrami and Page models. Secondly, the release rates of the ICs experienced an unsteady state in the early stage, and gradually became almost constants period after 20 hours. Except that the release of myrcene in γ-CD/myrcene belonged to the first-order kinetic, the release models of the remaining three ICs belonged to diffusion mode. Thirdly, the calculated binding energies of the optimized structures for α-CD/myrcene, ß-CD/myrcene, γ-CD/myrcene, and HP-ß-CD/myrcene ICs were -4.28, -3.82, -4.04, and -3.72 kcal/mol, respectively. Finally, the encapsulation of myrcene with α-CD and ß-CD was preferable according to the stability and release characteristics. CONCLUSION: The encapsulation of myrcene was profoundly affected by the type of CDs, and the stability could be improved by complexation with suitable CDs. The binding behavior between guest and CD molecules, and the release profile of the guest molecules could be effectively explained by the kinetics parameters and molecular modelling. This study can provide an effective basis and guide for screening suitable shell matrices.

Five new species of the genus Noeetomima Enderlein (Diptera: Lauxaniidae) from China, with a key to world species.

Five species of Noeetomima Enderlein, 1937 from Southwest China are described as new to science: Noeetomima hongshanensis sp. nov., N. lijiangensis sp. nov., N. liui sp. nov., N. trisurstyla sp. nov. and N. zhangae sp. nov.. A key to world species of the genus Noeetomima is presented. The specimens studied are deposited in the Entomological Museum of China Agricultural University, Beijing, China (CAUC).

[Preparation and in vitro quality evaluation of curcumin supersaturated self-nanoemulsion].

In order to improve the supersaturation and maintenance time of drug dispersion in curcumin self-nanoemulsion(CUR-SNEDDS), precipitation inhibitors(PPIs) were introduced to prepare curcumin supersaturated self-emulsion(CUR-SSNEDDS). The composition of CUR-SNEDDS prescriptions was selected through the solubility test, the compatibility of oil phase and surfactant, the investigation of the emulsifying ability of the surfactant and the drawing of the pseudo-ternary phase diagram. Analytic hierarchy process was used in combination with central composite design-response surface method to optimize the prescription. The type and dosage of precipitation inhibitors(PPIs) were selected to maintain the supersaturated concentration and duration of CUR in artificial gastrointestinal fluids. At the same time, polarizing microscope was used to evaluate the crystallization inhibition effect and the quality and in vitro release behavior of CUR-SSNEDDS. The prepared CUR-SSNEDDS prescription was capryol 90-kolliphor RH40-transcutol HP-Soluplus(7.93∶66.71∶25.36∶5), with the drug loading of(65.12±1.25) mg·g~(-1). CUR-SSNEDDS was transparent yellow, and the nanoemulsion droplets were spherical with uniform distribution. The emulsification time was(21.02±0.13) s, the average particle size was(57.03±0.35) nm, the polydispersity index(PDI) was(0.23 ± 0.01), and the Zeta potential was(-18.10±1.30) mV. CUR-SSNEDDS significantly inhibited the generation and growth of crystals after in vitro dilution. The supersaturation could be maintained above 10 within 2 h, and the dissolution rate and degree of CUR in artificial gastrointestinal fluid were significantly increased. Soluplus could effectively maintain the supersaturated state of CUR and enhance CUR dissolution in vitro.

Four new species of the subgenus Minettiella from China (Diptera, Lauxaniidae, Minettia).

Four species of Minettia Robineau-Desvoidy, 1830 from Southwest China are described as new to science: Minettia (Minettiella) dashahensissp. nov., M. (Minettiella) longispinasp. nov., M. (Minettiella) membranaceasp. nov. and M. (Minettiella) zhaotongensissp. nov. A key to the species of the subgenus Minettiella of the world is presented.

Aberrant methylation of IGF2-AS promoter in early pregnancy loss.

OBJECTIVE: The present study aimed to evaluate insulin-like growth factor 2 antisense (IGF2-AS) in the villi of human embryos and compared its expression between normal pregnancy and early pregnancy loss (EPL). MATERIALS AND METHODS: The present study conducted a microarray analysis to identify the expression profiles of lncRNAs in villi from EPL and normal controls (controls, n = 10 and EPL patients, n = 10). Embryonic villi were collected from women who underwent artificial abortion. QPCR was used to confirm the results. The DNA methylation patterns were analyzed using pyrosequencing and bisulfite sequencing polymerase chain reaction. The percentage of methylation was compared in chorionic villi from the two groups. RESULTS: A total of 57 deregulated differentially expressed lncRNAs were detected, of which 33 were upregulated, and 24 were downregulated. The expression of lncRNA IGF2-AS was downregulated significantly in EPL villi compared with the normal villi. Negative regulation of IGF2-AS may be involved in the development of EPL. Methprimer predicted that IGF2-AS promoter had CpG islands and dense CG sites. Increased methylation at CpG islands present in IGF2-AS gene promoter was observed in EPL villi. CONCLUSION: An increase in methylation of IGF2-AS likely leads to its downregulation in chorionic villi of EPL. The findings suggest that a deficiency of IGF2-AS in the villi is associated with human EPL.

A New Technique: Laparoscopic Resection of Fibrous Ring and Placing Extravascular Stent in Patients With Nutcracker Syndrome: A Report of 5 Cases.

OBJECTIVE: To evaluate the feasibility and safety of a new laparoscopic technique for resection of a fibrous ring and extravascular stent implantation in patients with nutcracker syndrome (NCS). MATERIAL AND METHODS: We retrospectively reviewed data of 5 patients diagnosed with NCS between March 2010 and February 2016. The mean age of the patients (4 male and 1 female) was 34 years (range, 28-40 years). All 5 patients underwent laparoscopic resection of the narrow fibrous ring around the left renal venous (LRV) and for extravascular stent implantation in the LRV for management of NCS. RESULTS: The average operating time was104 minutes and the average blood loss during surgery was 59 mL. The average length of the postoperative hospital stay was 6 days (range, 4-8 days). In all 5 patients, the symptoms of macroscopic hematuria started decreasing gradually and resolved after surgery. Postoperative computed tomography showed that the blood outflow from the LRV was smooth. The ratio of the dilated segment's inner diameter to the diameter of the strictured segment decreased from 3.4 to 9.5, preoperatively to 1.1-2.0, postoperatively. The mean follow-up period was 17.6 months (range, 8-24 months).One patient's varicocele was cured and symptoms in all 5 patients resolved after surgery. None of the patients showed symptom recurrence. CONCLUSION: Laparoscopic surgery, for the placement of an extravascular stent and resection of the fibrous ring around the end of the LRV outflow to the inferior vena cava appears feasible and safe and offers an alternative minimally invasive for the management of NCS.

Preparation, Characterization, and Pharmacokinetic Evaluation of Imperatorin Lipid Microspheres and Their Effect on the Proliferation of MDA-MB-231 Cells.

Imperatorin is a chemical compound belonging to the linear furanocoumarins. Imperatorin is attracting considerable attention because of its antitumor, antibacterial, anti-inflammatory, and anticoagulant activities, inhibition of myocardial hypertrophy, and other pharmacological efficacies. However, imperatorin has limited water solubility and has better lipid solubility; thus, we decided to design and synthesize imperatorin lipid microspheres to optimize the preparation conditions. The aim was to develop and formulate imperatorin lipid microspheres through nanoemulsion technology and apply the response surface⁻central composite design to optimize the imperatorin lipid microsphere formulation. The influence of the amounts of egg lecithin, poloxamer 188, and soybean oil for injection on the total percentage of the oil phase was investigated. The integrated effect of dependent variables, including particle size, polydispersity index, zeta potentials, drug loading, and encapsulation efficiency, was investigated. Data of overall desirabilities were fitted to a second-order polynomial equation, through which three-dimensional response surface graphs were described. Optimum experimental conditions were calculated by Design-Expert 8.06. Results indicated that the optimum preparation conditions were as follows: 1.39 g of egg lecithin, 0.21 g of poloxamer 188, and 10.57% soybean oil for injection. Preparation of imperatorin lipid microspheres according to the optimum experimental conditions resulted in an overall desirability of 0.7286, with the particle size of 168 ± 0.54 nm, polydispersity index (PDI) of 0.138 ± 0.02, zeta potentials of -43.5 ± 0.5 mV, drug loading of 0.833 ± 0.27 mg·mL-1, and encapsulation efficiency of 90 ± 1.27%. The difference between the observed and predicted values of the overall desirability of the optimum formulation was in the range from 2.4% to 4.3%. Subsequently, scanning electron microscopy was used to observe the micromorphology of the imperatorin lipid microspheres, showing round globules of relatively uniform shape and sizes within 200 nm. The effect of imperatorin lipid microspheres on MDA-MB-231 proliferation was investigated by the MTT method. Furthermore, pharmacokinetics in Sprague-Dawley rats was evaluated using orbital bleeding. A sensitive and reliable liquid chromatography with the high-performance liquid chromatography (HPLC) method was established and validated for the quantification of imperatorin in rat plasma samples. The data were calculated by DAS (drug and statistics) Pharmacokinetic Software version 3.3.0 (Version 3.3.0, Shanghai, China). Results demonstrated that imperatorin lipid microspheres can significantly enhance the bioavailability of imperatorin and can significantly inhibit MDA-MB-231 cell proliferation. In conclusion, our results suggested that the response surface⁻central composite design is suitable for achieving an optimized lipid microsphere formulation. Imperatorin lipid microspheres can improve the bioavailability of imperatorin and better inhibit the proliferation of MDA-MB-231 cells as compared to imperatorin alone.

Expression of TET and 5-HmC in Trophoblast Villi of Women with Normal Pregnancy and with Early Pregnancy Loss.

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy. The ten-eleven translocation (TET) family and 5-hydroxymethylcytosine (5-hmC) were found to be linked with epigenetic reprogramming. The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss (EPL). Embryonic villi were collected from normal pregnant women (control) experiencing medical abortion and from EPL patients at gestation ages of 6, 7 and 8 weeks. The mRNAs of TET family were analysed using quantitative polymerase chain reaction (qPCR), and TET proteins using Western blotting and immunohistochemical analysis. The MethylFlash™ Kit was used to quantify the absolute amount of 5-methylcytosine (5-mC) and 5-hmC. Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age. Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples, which was consistent with the qPCR and Western blot results. The expression of TET1, TET2, and TET3 was lower in the villi in EPL group than in normal pregnancy group (P<0.05 for all). It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo. The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.