Bacterial, short-chain fatty acid and gas profiles of partially hydrolyzed guar gum in vitro fermentation by human fecal microbiota.

Carbohydrates with different structures have metabolic differences in the human body, as well as individual differences. The present study aimed to investigate the effects of bacterial, short-chain fatty acids (SCFAs) and gas profiles of partially hydrolyzed guar gum (PHGG) on the fecal microbiota of 41 Chinese individuals by simulated fermentation in vitro. Results showed that PHGG stimulated the growth of Bifidobacterium and Faecalibacterium, inhibited the growth of Escherichia-Shigella, Klebsiella, and Dorea, and induced the production of fermentation gases (CO2, and H2) and SCFAs (acetic acid, butyric acid). Furthermore, Bifidobacterium was significantly increased in the young female and the old male-originated samples, while Klebsiella was significantly decreased in the old female ones after PHGG intervention, and there were also certain differences in gases and SCFAs among different population samples. These findings indicate that PHGG can modulate gut microbiota and metabolism well, whereas its use varies in different populations.

Dietary supplementation with synthetic astaxanthin and DHA interactively regulates physiological metabolism to improve the color and odor quality of ovaries in adult female Eriocheir sinensis.

The present study aimed to investigate the interactive effects of dietary supplementation to the feed with astaxanthin and/or DHA on the color and odor of Eriocheir sinensis ovaries. The results revealed that astaxanthin supplementation significantly increased redness of E. sinensis ovaries (P < 0.05). Moreover, the addition of either astaxanthin or DHA alone to the feed affected the deposition of carotenoids and fatty acids in E. sinensis ovaries. More importantly, the simultaneous supplementation of astaxanthin and DHA significantly improved color, carotenoid content, and polyunsaturated fatty acid content (P < 0.05) in E. sinensis ovaries, as well as increased the content of aroma compounds during thermal processing. Based on the present findings, the optimal combination of dietary astaxanthin and DHA is 100 mg/kg of synthetic astaxanthin and 0.15% of DHA, respectively, which could improve color and odor quality of ovaries for E. sinensis.

Label-Free quantitation of milk oligosaccharides from different mammal species and heat treatment influence.

Milk oligosaccharides (MOs) exhibit significant variations in concentrations and patterns among different species. However, there is limited knowledge about milk oligosaccharides in domestic animals and the impact of heat treatment on them. Here, we developed an LC-ESI-MS/MS method to analyze 11 milk oligosaccharides in 7 distinct species simultaneously. The results showed that human milk presented a completely different composition pattern of milk oligosaccharides from animals. In detail, animal milk predominantly contained sialylated oligosaccharides, and human milk had high levels of fucosylated neutral oligosaccharides. Notably, sheep milk exhibited similarities to human milk in terms of oligosaccharides composition. Then, the milk samples from dairy cows were treated with two common industrial heat treatments. We found that 65 °C treatment had no significant effect on the concentration of milk oligosaccharides, whereas 135 °C heating was associated with their decline, suggesting that high temperatures should be avoided in the processing of oligosaccharides supplemented/enriched products.

Active help-seeking and metacognition interact in supporting children's retention of science facts.

Determining when to ask for help is a critical self-regulated strategy that can benefit children's learning. Despite its importance, we have a limited understanding about the developmental mechanisms that support adaptive help-seeking. In the current preregistered study, predominately White children aged 8 to 13 years (N = 69, ngirls = 37) had the option to seek help during an online science learning task. Results revealed that children's ability to adaptively seek help improved throughout childhood and early adolescence. Critically, developing metacognitive skills contributed to greater help-related memory benefits (compared with conditions where help was not previously available). Overall, these findings highlight the role of metacognition in children's ability to adaptively seek and benefit from help during online science learning.

1D-, 2D-Gel Electrophoresis, Immunoblotting, and Enzyme-Linked Immunosorbent Assay (ELISA) for the Study of Food Allergens.

Protein-based methods have been fundamental for the study of food allergens, not only from a mechanistic and diagnostic point of view, but especially with respect to allergen management and food safety. In this chapter, four individual protocols are suggested, relying on one-dimensional, two-dimensional gel electrophoresis, immunoblotting, and enzyme-linked immunosorbent assay (ELISA). The particularities of the proposed protocols are focused on previous research targeting specific allergenic foods, with cow's milk proteins as case studies. Data on the importance of protein extraction and the use of different animal-raised antibodies and/or sera of food-allergic patients are also critically discussed within method development and optimization. The protocols herein described are successful examples applied to the study of cow's milk allergens in complex matrices, although they can be easily developed and optimized for any food allergen or allergenic food.

Translation, cross-cultural adaptation, and validation of health and self-management in diabetes questionnaire (HASMID-10) into Brazilian Portuguese.

BACKGROUND: Considering the ability of the health and self-management in diabetes questionnaire (HASMID-10) to verify the impact of self-management on diabetes, we highlight its relevance to scientific research and clinical applicability. However, to date, no study has been conducted to scientifically support its use in other languages. OBJECTIVE: To translate, cross-culturally adapt, and validate the HASMID-10 into the Brazilian Portuguese. DESIGN AND SETTING: A translation, cross-cultural adaptation, and validation study conducted at Ceuma University. METHODS: Study was conducted in accordance with the Guidelines for the Process of Cross-Cultural Adaptation of Self-Report Measures and Consensus-based Standards for the Selection of Health Measurement Instruments. We included participants of both sexes diagnosed with diabetes, aged between 18 and 64 years, and without cognitive deficits or any other limitations that would prevent them from answering the questionnaire. We assessed participants using the problem areas in diabetes (PAID) scale and HASMID-10. We assessed reliability using a test-retest model with a 7-day interval between assessments. We used intraclass correlation coefficient (ICC), 95% confidence interval (CI), standard error of measurement (SEM), minimum detectable difference (MDD), Spearman correlation coefficient, and floor and ceiling effects. RESULTS: Sample comprised 116 participants, most of whom were women, overweight, non-practitioners of physical activity, and nonsmokers. We observed significant correlations (P = 0.006; rho = -0.256) between the HASMID-10 and PAID, adequate reliability (ICC = 0.780) and internal consistency (Cronbach's alpha = 0.796). No ceiling or floor effects were observed. CONCLUSION: HASMID-10 has adequate measurement properties and may be used for Brazilians.

Sonocatalytic degrading antibiotics over activated carbon in cow milk.

Herein, an efficient, simple and economical approach to remove antibiotics (ABX), i.e. ceftiofur hydrochloride, sulfamonomethoxine sodium (SMM), marbofloxacin and oxytetracycline by sonication with activated carbon (AC) from cow milk has been successfully implemented. The pseudo-first-order kinetics constants for the sonolytic and sonocatalytic degrading SMM are 0.036 and 0.093 min-1, respectively. The synergistic efficiency of removing ABX by using sonocatalysis reached 1.8-4.0. Hydrophobic ABX underwent faster degradation than hydrophilic ABX in sonocatalytic systems. Adding 0.5 mmol L-1 H2O2, the optimal concentration, improved the sonocatalytic degradation rates of ABX by 9.1%-28.5%. Surface area and dose of AC play crucial roles in the sonocatalysis of ABX. By sonicating 50 mL of 5.52 µmol L-1 ABX in milk at 500 kHz and 259 W with 20 mg AC for 20-60 min resulted in residual ABX concentrations ranging from 42.6 to 95.1 µg L-1, which meet the relative maximum residue limits set by European Commission.

Trophoblast organoids with physiological polarity model placental structure and function.

Human trophoblast organoids (TOs) are a three-dimensional ex vivo culture model that can be used to study various aspects of placental development, physiology and pathology. However, standard culturing of TOs does not recapitulate the cellular orientation of chorionic villi in vivo given that the multi-nucleated syncytiotrophoblast (STB) develops largely within the inner facing surfaces of these organoids (STBin). Here, we developed a method to culture TOs under conditions that recapitulate the cellular orientation of chorionic villi in vivo. We show that culturing STBin TOs in suspension with gentle agitation leads to the development of TOs containing the STB on the outer surface (STBout). Using membrane capacitance measurements, we determined that the outermost surface of STBout organoids contain large syncytia comprising >50 nuclei, whereas STBin organoids contain small syncytia (<10 nuclei) and mononuclear cells. The growth of TOs under conditions that mimic the cellular orientation of chorionic villi in vivo thus allows for the study of a variety of aspects of placental biology under physiological conditions.

All-in-one coupling of 3D hybridized nanocarbon microelectrode for portable monitoring of doxycycline hyclate.

The simultaneous balance of electrode materials and electrode structures can energize the development of innovative electrochemical sensors. In this work, a 3D nanocarbon layer of hybrid heteroatoms and metal atoms (CN/Fe) with excellent electrical properties and abundant active sites was self-constructed on the surface of a quartz-based nanofiber by high-temperature pyrolysis. Further, the nanofiber tip was selected as the sensing region to develop an electrochemical sensing platform with high sensitivity, miniaturization, and portability. A common broad-spectrum antibiotic (Doxycycline hyclate, DOX) was used as a model to evaluate the designed miniaturized sensing platform, and the stability, reproducibility, and applicability of the microsensor were verified in a variety of real samples, including algal solution, milk, human serum, and cell culture media. The results show that the proposed sensing platform has a detection limit as low as 82 nM in aqueous environments. Furthermore, it is further shown that coupling the design of electrode materials and electrode structures facilitates the development of electrochemical sensors with more practical applications. This concept will open up new avenues for the development of electrochemical sensors that meet many application scenarios.

An eco-friendly sample preparation procedure based on air-assisted liquid-liquid microextraction for the rapid determination of phthalate metabolites in urine samples by liquid chromatography-tandem mass spectrometry.

Urinary phthalate metabolite (mPAEs) analysis is a reliable tool for assessing human exposure to phthalates. With growing interest in urinary biomonitoring of these metabolites, there is a need for fast and sensitive analytical methods. Therefore, a simple, rapid procedure for simultaneous determination of fifteen phthalate metabolites in human urine samples by liquid chromatography-tandem mass spectrometry was developed. The novelty of the present procedure is based on the use of diethyl carbonate as a green biobased extraction solvent and air-assisted liquid-liquid microextraction (AALLME) as a sample preparation step. A Plackett-Burman design was used for screening the factors that influence the AALLME extraction efficiency of mPAEs. The effective factors were then optimized by response surface methodology using a central composite rotatable design. Under the optimized conditions, good linearity can be achieved in a concentration range of 1.0-20.0 ng mL-1 with correlation coefficients higher than 0.99. The repeatability and reproducibility precision were in the range of 2-12% and 1-10% respectively. Recoveries ranging from 90% to 110%. This, and the low limits of detection, ranging from 0.01 to 0.05 ng mL-1, make the proposed procedure sensitive and suitable for human biomonitoring of phthalate exposures. For proof-of-principle, the new method was used to measure the urinary concentrations of mPAEs in 20 urine samples from Brazilian women. The high frequency of detections and in part high concentrations of mPAEs indicate to widespread exposure to several phthalates among Brazilian women.

High throughput LC-MS/MS method for steroid hormone analysis in rat liver and plasma - unraveling methodological challenges.

Comprehensive reference data for steroid hormones are lacking in rat models, particularly for early developmental stages and unconventional matrices as the liver. Therefore, we developed and validated an enzymatic, solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify a panel of 23 steroid hormones in liver and plasma from adult and neonatal rats. Our approach tackles methodological challenges, focusing on undesired byproducts associated with specific enzymatic treatment, and enables a thorough assessment of potential interferences in complex matrices by utilizing unstripped plasma and liver. We propose an optimized enzymatic hydrolysis protocol using a recombinant ß-glucuronidase/sulfatase mix (BGS mix) to efficiently deconjugate steroid phase II conjugates. The streamlined sample preparation and high-throughput solid phase extraction in a 96-well plate significantly accelerate sample processing for complex matrices and alarge number of samples. We were able to achieve the necessary sensitivity for accurately measuring the target analytes, particularly estrogens, in small sample sizes of 5-20 mg of liver tissue and 100 µL of plasma. Through the analysis of liver and plasma samples from adult and neonatal rats, including both sexes, our study showed a novel set of steroid hormone reference intervals. This study provides a reliable diagnostic tool for the quantification of steroids in rat models and gives insight in liver and plasma-related steroid hormone dynamics at early developmental stages. In addition, the method covers several pathway intermediates and extend the list of steroid hormones to be investigated.

Beneficial biological effects of Flavokawain A, a chalcone constituent from kava, on surgically induced endometriosis rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: Shrub kava has long been grown and utilized, primarily in the South Pacific region, for ceremonial, religious, and social occasions. It has been used as a pain reliever and muscle relaxant in medicinal practices from the eighteenth century. Interestingly, relatively low incidence of lung cancer may attribute to the high consumption of kava products in this region. AIM OF THE STUDY: Kava extracts were used to produce the kava chalcones Flavokawain A, B and C, which have a variety of bioactivities. In the present study, we show that Flavokawain A has positive effects on endometriosis. MATERIALS AND METHODS: The endometriosis rat model was surgically induced by the autologous transplantation of endometrial tissue. Rats were evaluated for clinical ratings and lesion volume following a 6-week Flavokawain A therapy. Peritoneal fluid and blood samples were taken and ELISA assay was used to measure the cytokines and chemokines levels. Transcriptional and expression levels of Akt, PI3K, NF-kB, iNOS, Bcl-2, Bax and caspase-3 were evaluated by Western blotting and RT-qPCR. Implanted tissue sections of the rats were also analyzed by immunofluorescent and histopathological staining. RESULTS: Lesion volumes and adhesion scores were successfully decreased. Blood and peritoneal fluid levels of associated cytokines and chemokines were markedly down-regulated. Besides, Flavokawain A also mediated cell apoptosis of endometrial implants. Additionally, VEGF expression was reduced, which inhibited the angiogenesis process. As for the expression of Akt, p-Akt, PI3K, p-PI3K, and NF-kB in endometriosis lesions, Flavokawain A significantly reduced them. CONCLUSION: Flavokawain A has beneficial effects on the surgically induced endometriosis rat model, by reducing inflammation, promoting apoptosis, and decreasing angiogenesis. Our findings suggest that these effects may be mediated through the regulation of PI3K/Akt and NF-κB signaling pathways.

Curcuma latifolia Roscoe extract reverses inflammatory pain in mice and offers a favorable CNS safety profile.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma latifolia Roscoe, a plant in the Curcuma genus, has been used as a food additive and folk medicine in Thailand to treat pelvic pain and improve premenstrual syndrome. Although it has been used for centuries, no scientific studies have proved its potential effects on inflammatory pain and central nervous system (CNS) safety profiles. AIM OF THE STUDY: This study aimed to evaluate the potential effects of the ethanolic extract of C. latifolia rhizome on inflammatory pain in mice, together with its CNS safety profiles. MATERIALS AND METHODS: First, network pharmacology was employed to identify the role of bioactive constituents in C. latifolia on inflammatory pain. In addition, in vitro pharmacology was also evaluated to confirm the anti-inflammatory activity of C. latifolia extract at cellular levels in activated macrophages and microglia. Furthermore, the efficacy of the plant extract in attenuating formalin-induced pain-like behaviors in mice was evaluated. Mice were orally administered the extract (125, 250, 500 mg/kg) followed by the measurement of formalin-induced pain-like behaviors. The LABORAS automated behavioral analysis and rotarod test were used to assess potential CNS side effects of C. latifolia extract (500 mg/kg) in mice. RESULTS: The results demonstrated that major bioactive constituents present in C. latifolia have the ability to regulate multiple targets, biological processes and pathways associated with inflammatory pain as assessed by network pharmacology. C. latifolia modulated peripheral and central immune cells via reducing proinflammatory mediators (NO, TNF-α, and IL-6). C. latifolia extract improved formalin-induced pain-like behaviors in a dose-dependent manner during phase II of the formalin test. The efficacy of the plant extract at doses of 250 and 500 mg/kg was comparable to that of the positive control (indomethacin 10 mg/kg). Furthermore, the highest therapeutic dose of the extract did not affect motor coordination, exploratory behaviors, general behaviors, and overall well-being of mice, indicating no development of potential CNS adverse effects after administration of the extract. CONCLUSION: These findings provide novel perspectives on using C. latifolia extract for pain management, considering its therapeutic efficacy and CNS safety.

Toxicological safety evaluation of zengye granule through acute and 30-day toxicity studies in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Zengye granule (ZYG), a traditional Chinese medicine formula composed of Radix Scrophulariae, Radix Ophiopogonis, and Radix Rehmanniae in the ratio of 1.0:0.8:0.8, is listed in the Chinese Pharmacopoeia for treating diseases associated with yin deficiency, such as inner heat, dry mouth and pharynx, and dry bound stool. However, little information is available on its toxicological safety. AIM OF THE STUDY: To evaluate the acute and subacute toxicity of ZYG after oral administration in rats. MATERIALS AND METHODS: In the acute toxicity study, ZYG was orally administered to rats at a single dose of 10 g/kg/day. In the subacute toxicity study, ZYG was administered orally to rats at repeated daily doses of 2.5, 5.0, or 10 g/kg/day for 30 days. The toxicological effects were evaluated by assessing the rats' general behavior, body weight, food intake, water consumption, blood biochemical and hematological parameters, organ coefficients, and organ histopathology. RESULTS: No obvious adverse reactions were found in the rats in the acute toxicity study, indicating that ZYG was non-toxic. In the subacute toxicity study, ZYG had no toxic effect on the rats at a dose of 2.5 g/kg/day but showed slight toxicity in the kidneys, and spleens of the rats at doses of 5 and 10 g/kg/day. Significant drug toxicity was observed in male and female rats at 5 and 10/kg/day; however, elevated WBCs counts, ALT, and LYMs levels were found in female rats. CONCLUSIONS: The oral administration of ZYG at a dose of less than 10 g/kg/day for 1 day or 2.5 g/kg/day for 30 consecutive days can be considered safe, as these doses showed no distinct toxicity or side effects in the rats in this study. Therefore, the dosage should be set according to the clinically recommended dosage to ensure its safety.

Jiawei Shoutai Pill promotes decidualization by regulating the SGK1/ENaC pathway in recurrent spontaneous abortion.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiawei Shoutai Pill (JWSTW) is a traditional herbal formula for recurrent spontaneous abortion (RSA). Although JWSTW significantly improves the clinical symptoms of RSA patients, its molecular mechanism remains unclear. AIM OF STUDY: This study evaluated the expression and function of the serum/glucocorticoid regulated kinase 1/epithelial sodium channel (SGK1/ENaC) pathway and decidualization level in RSA patients and mice. It also investigated the therapeutic effects and potential mechanisms of JWSTW. MATERIALS AND METHODS: 30 early RSA patients and 30 normal pregnant women undergoing induced abortion during the same period were included in the study. Decidual tissues were collected, and HE staining, immunohistochemistry, Western blot, and RT-PCR were used to detect protein and mRNA expression levels of SGK1, ENaC-a, estrogen Rreceptor ß (ERß), and progesterone receptor (PR) in patients' decidual tissues. Protein expression levels of prolactin receptor (PRLR) and insulin-like growth factor binding protein 1 (IGFBP-1) were also detected. A classical RSA mouse model was constructed, and the mice were randomly divided into four groups: normal, model, dydrogesterone (DQYT) (0.33 g/kg/d), and JWSTW (1.66 g/kg/d). The normal and model groups received the same volume of distilled water by gavage for 8 and 14 days after pregnancy. On the 14th day of pregnancy, the embryonic loss rate of each group, the number of offspring born to naturally delivered mice, and the protein or mRNA expression levels of key factors of the SGK1/ENaC pathway (SGK1, ENaC-a, ERß, and PR), decidual proliferation marker (Ki67), mesenchymal-epithelial transition (E-cadherin and Vimentin), and decidualization markers (PRLR and IGFBP-1) in mouse decidual tissue on the eighth day of pregnancy were observed. RESULTS: The decidual tissue structure of RSA patients was abnormal. Immunohistochemical analysis revealed significantly reduced positive expression of SGK1, ENaC-a, ERß, and PR proteins in the decidual tissue of RSA patients (P < 0.001). Western blot and RT-PCR analyses demonstrated significantly decreased protein and mRNA expression of SGK1, ENaC-a, ERß, and PR in the decidual tissue of RSA patients (all P < 0.05). Additionally, protein expression of PRLR and IGFBP-1 was significantly reduced (both P < 0.001). The RSA mouse model exhibited a significant increase in embryo loss rate and decreased litter size (both P < 0.001). Treatment with DQYT and JWSTW rescued the embryo loss rate and litter size to varying extents (all P < 0.05). The protein or mRNA expression levels of SGK1, ENaC-a, ERß, PR, Ki67, E-cadherin, vimentin, PRLR, and IGFBP-1 in RSA mice were improved to different degrees after treatment with DQYT and JWSTW (all P < 0.05). CONCLUSIONS: Abnormal SGK1/ENaC signaling pathway regulation is closely associated with early endometrial damage in RSA patients. JWSTW promotes endometrial proliferation and mesenchymal-epithelial transition through the SGK1/ENaC signaling pathway, improving endometrial shedding. Consequently, JWSTW is a potential treatment for RSA.

In view of ovarian steroidogenesis and luteal construction to explore the effects of Bushen Huoxue recipe in mice of ovarian hyperstimulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Bushen Huoxue recipe (BSHXR) is a widely used prescription medicine for treating gynecological diseases. We have previously found that BSHXR can improve the pregnancy outcome of controlled ovarian hyperstimulation (COH) mice by modulating the abnormal high level of progesterone. While the pharmacological mechanism of such therapeutic effect is not clear. AIM OF THE STUDY: We aimed to investigate the effects of BSHXR on the ovarian steroidogenesis and luteal function in mice undergoing COH. MATERIALS AND METHODS: A COH mouse model was established via an intraperitoneal injection of 0.4 IU/g pregnant mare serum gonadotropin (PMSG) and 1 IU/g human chorionic gonadotropin (HCG). The histological features of ovaries were observed using hematoxylin-eosin staining. The expression levels of FSHR, LHCGR, and key molecules in ovarian steroidogenesis, including CYP11A1, CYP17A1, CYP19A1, HSD3B1, and StAR, were examined via immunohistochemical staining, western blotting, and RT-qPCR. CD31, VEGFA, and FGF2 levels were assessed to evaluate ovarian vascularization. The protein and mRNA levels of ovarian ERK1/2, p-ERK1/2, MEK1/2, and p-MEK1/2 were also detected using western blotting, RT-qPCR, or immunofluorescence staining. RESULTS: COH mice had a significantly increased volume and weight of the ovary and number of corpora lutea. In particular, COH exhibited a long-term influence on ovarian FSHR and LHCGR expression, disrupting the levels of CYP11A1, HSD3B1, and CYP17A1, causing poorer luteal angiogenesis. Compared with normal mice, the expression levels of ovarian VEGFA and FGF2 in COH mice were considerably lower on Day 1 after PMSG. On concomitant HCG treatment, both VEGFA and FGF2 expression surged dramatically on ED1 and then declined on ED4 and ED8. Moreover, the expression pattern of MEK1/2-ERK1/2 was almost consistent with that of VEGFA and FGF2. After treatment, BSHXR increased ovarian LHCGR, FSHR, CYP11A1, HSD3B1, and CYP17A1 levels, boosted luteal vascularization, and restored MEK1/2-ERK1/2 signaling in COH mice. CONCLUSION: BSHXR restored the abnormally high progesterone level by regulating the CYP11A1 and HSD3B1 expression as well as promoted luteal angiogenesis, which was related with LHCGR-MEK1/2-ERK1/2-VEGFA/FGF2 signaling pathway in the ovary. This effect prevented the fluctuation of sex hormones in COH mice and benefited the outcome of pregnancy.

Pingchong Jiangni recipe through nerve growth factor/transient receptor potential vanilloid 1 signaling pathway to relieve pain in endometriosis model rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Pingchong Jiangni recipe (PJR) is often used in the treatment of endometriosis (EM). This formula has been clinically validated by the State Administration of Traditional Chinese Medicine Key Specialties Collaborative Group for its therapeutic efficacy. Recently, our research team also confirmed that PJR has a shrinking effect on ovarian chocolate cysts. Additionally, PJR was also found to have a shrinking effect on EM lesions; however, the mechanism by which this effect occurs remains unclear. AIM OF THE STUDY: To explore the mechanisms by which PJR relieves pain in patients with EM. MATERIALS AND METHODS: A rat model of EM was established by autologous transplantation. PJR (3.78 g/kg, 7.56 g/kg, and 15.12 g/kg) was orally administered for 21 days. The rat grimace scale (RGS) score and paw withdrawal threshold (PWT) were measured at a fixed time during the experiment. Hematoxylin and eosin staining was performed to observe histopathological changes in EM rats after administration, enzyme-linked immunosorbent assay to evaluate the plasma expression of tumor necrosis factor-α (TNF-α) and nerve growth factor (NGF), and immunohistochemistry and western blotting to identify differences in the expression of pain-related factors in EM rats. RESULTS: The medium-dose group of PJR (7.56 g/kg) had the best effect on relieving pain in EM rats by reducing RGS, increasing PWT, reducing the ectopic endometrium, improving the cellular structure of the lesion, and reducing TNF-α and NGF levels. However, PJR significantly decreased the expression of transient receptor potential vanilloid 1 (TRPV1), phosphorylated TRPV1 (p-TRPV1), protein kinase C (PKC), and NGF. CONCLUSION: The mechanism by which PJR relieves EM pain may be through the downregulation of NGF, PKC, and TRPV1 expression.

The effect of an oral product containing Amla fruit (Phyllanthus emblica L.) on female androgenetic alopecia: A randomized controlled trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Amla (Phyllanthus emblica) fruit has been emphasized as a hair tonic in Traditional Persian Medicine (TPM) and recommended for hair loss orally and topically. AIM OF THE STUDY: This study aimed to investigate the effect of an oral product containing Amla fruit on Female Androgenetic Alopecia (FAGA). MATERIALS AND METHODS: This study was a triple-blind, randomized, controlled clinical trial. Sixty women with FAGA were randomly assigned into two groups of thirty. The intervention group received ten cc Amla syrup thrice a day for 12 weeks. The second group received a placebo with the same dose and duration. Hair growth parameters were analyzed using TrichoScan before and after 12 weeks of intervention. Physician and patient satisfaction were assessed using the CGI-I and PGI-I questionnaires, respectively. RESULTS: Twenty-seven participants in the intervention group and 25 in the placebo group completed the trial. Based on our findings, the anagen-to-telogen ratio increased significantly in the intervention group compared with the group who received placebo (F = 10.4, P = 0.002). Physician and patient satisfaction increased in the amla group compared with placebo at 12th weeks of intervention (P<0.001), (P<0.001). The formula had no remarkable side effects. Only one case of mild constipation was reported in one of the participants after one month of consuming Amla syrup. CONCLUSION: The results of this study demonstrated that Amla syrup could help treat androgenic hair loss in women and increase the anagen phase. Further studies are needed to evaluate this potential treatment for FAGA.

Inhibitory effect of Lonicera japonica-derived exosomal miR2911 on human papilloma virus.

ETHNOPHARMACOLOGICAL RELEVANCE: Lonicera japonica Thunb. has been used as a traditional medicinal herb in China for thousands of years for its heat-clearing and detoxification effects. In recent years, experimental and clinical studies have shown that some Lonicera japonica-containing Chinese medicine prescriptions have been used to treat intraepithelia neoplasia caused by human papilloma virus (HPV) infection. However, its bioactive molecules and mechanism of action have not been fully explored. AIM OF THE STUDY: In this study, Lonicera japonica-derived exosomes was extracted and exosomal miR2911 was identified. Bioinformatic analysis indicated that miR2911 potentially binds to the sequence of HPV. The mechanism of miR2911 action on HPV and the effect of exosomal miR2911 on HPV-induced cervical cancer cells were investigated. METHODS: The potential targets of miR2911 on the HPV sequence were predicted and confirmed by using RNAhybrid and dual-luciferase reporter assays. Lonicera japonica exosomes were characterized by transmission electronic microscopy and zeta sizer analysis. RT-qPCR was used to measure miR2911 concentration in various tissues and exosomes. Synthetic miR2911 and GFP-E6/E7 plasmids were transfected into HEK293T cells to examine the effect of miR2911 on E6/E7 gene expression. The effects of miR2911 on endogenous E6/E7 mRNA and protein levels were detected in HPV16/18-positive cervical cancer cells by RT-qPCR and Western blotting. The proliferation and apoptosis of CaSki, SiHa and HeLa cells by the treatment of miR2911 or miR2911-containing exosomes were examined by CCK8, colony formation and flow cytometry assays. RESULTS: MiR2911 is found to be enriched in various Lonicera japonica tissues, and is stably present in Lonicera japonica-derived exosomes. It is observed that MiR2911 directly binds to E6 and E7 oncogenes of HPV16/18, leading to the suppression of their protein expression. In addition, the endogenous E6/E7 mRNA and protein levels were significantly decreased by using miR2911 treatment in HPV16/18-positive cervical cancer cells. Furthermore, both miR2911 and miR2911-containing exosomes inhibited cell proliferation of SiHa, CaSki and HeLa cells, meanwhile inducing the cell apoptosis through E6/E7-p53/Caspase3 axis. CONCLUSION: Our findings indicate that miR2911, an active component present in Lonicera japonica exosomes, inhibits proliferation and induces apoptosis of cervical cancer cells by targeting the E6/E7 genes of HPV16/18. Thus, Lonicera japonica-derived exosomal miR2911 has implications for the development of novel therapeutic strategies for the treatment of HPV-associated cervical cancers.

Effects of BXSMD on ESR1 and ESR2 expression in CSD female mice.

ETHNIC PHARMACOLOGICAL RELEVANCE: Due to the rapid pace of modern society, chronic insomnia has become universal phenomenon. In China, Banxia Shumi Decoction (BXSMD) has been used in treating chronic insomnia for thousands of years, but its chemical composition and action mechanism are still unknown. AIM OF THE STUDY: This study aims to explore the chemical composition of BXSMD and its effects on Estrogen receptor 1 (ESR1) and Estrogen receptor 2 (ESR2) in mice with chronic sleep deprivation (CSD). MATERIALS AND METHODS: UHPLC-Q-Orbitrap-MS/MS was applied in determining the chemical composition of BXSMD. After 21-day sleep deprivation (SD) in platform water environment, CSD mice model was prepared. By conducting open field test, 24-h autonomic diurnal and nocturnal activity of mice in each group was detected. ELISA was employed to measure the contents of 5-HT, DA, NE, GABA, Glu, and MT. With RT-PCR, Western blot (WB), and immunohistochemistry (IHC), mRNA and protein expressions of ESR1 and ESR2 in the hypothalamus and hippocampus were tested. RESULTS: BXSMD included ferulic acid, kaverol, daidzein, apigenin, berberine, adenosine, aesculin, vanillin, naringin, and glycine, which might constitute the material basis forthe improvement of chronic insomnia. With BXSMD, the total moving distance and the rest time in dark period of CSD mice were shortened, while its rest time in light period was increased. Besides, BXSMD enhanced the contents of 5-HT, GABA, and MT in CSD mice, and decreased the contents of Glu, NE, and DA. BXSMD elevated the mRNA of Esr1 and Esr2, and elevated the protein expressions of ESR1 and ESR2 in the hypothalamus and hippocampus of CSD mice. CONCLUSIONS: BXSMD contains various chemical components for sleep-wake regulation, with the mechanism of stimulating estrogen signaling pathway by regulating the expressions of ESR1 and ESR2, ultimately realizing the regulation to sleep-wake disorder caused by CSD.