Preparing a novel baicalin-loaded microemulsion-based gel for transdermal delivery and testing its anti-gout effect.

We previously demonstrated that baicalin had efficacy against gouty arthritis (GA) by oral administration. In this paper, a novel baicalin-loaded microemulsion-based gel (B-MEG) was prepared and assessed for the transdermal delivery of baicalin against GA. The preparation method and transdermal capability of B-MEG was screened and optimized using the central composite design, Franz diffusion cell experiments, and the split-split plot design. Skin irritation tests were performed in guinea pigs. The anti-gout effects were evaluated using mice. The optimized B-MEG comprised of 50 % pH 7.4 phosphate buffered saline, 4.48 % ethyl oleate, 31.64 % tween 80, 13.88 % glycerin, 2 % borneol, 0.5 % clove oil and 0.5 % xanthan gum, with a baicalin content of (10.42 ± 0.08) mg/g and particle size of (15.71 ± 0.41) nm. After 12 h, the cumulative amount of baicalin permeated from B-MEG was (672.14 ± 44.11) µg·cm-2. No significant skin irritation was observed following B-MEG application. Compared to the model group, B-MEG groups significantly decreased the rate of auricular swelling (P < 0.01) and number of twists observed in mice (P < 0.01); and also reduced the rate of paw swelling (P < 0.01) and inflammatory cell infiltration in a mouse model of GA. In conclusion, B-MEG represents a promising transdermal carrier for baicalin delivery and can be used as a potential therapy for GA.

Baicalin inhibits monosodium urate crystal-induced pyroptosis in renal tubular epithelial cell line through Panx-1/P2X7 pathways: Molecular docking, molecular dynamics, and in vitro experiments.

Pyroptosis is a programmed cell death process that frequently occurs in many diseases, including hyperuricemic nephropathy (HN). In HN, a range of stimuli mediates inflammation, leading to the activation of inflammasomes and the production of gasdermin D (GSDMD). Baicalin (BA), a natural flavonoid renowned for its antioxidant and anti-inflammatory properties, was investigated for its role in HN in this study. Initially, HN-like inflammation and pyroptosis were induced in HK-2 cells with treatment of monosodium urate (MSU), followed by the BA treatment. The expression of pyroptosis-associated genes, Panx-1 and P2X7, at both mRNA and protein levels was assessed through real-time polymerase chain reaction (RT-qPCR) and Western blotting (WB) without or with BA treatment. The results showed that expression of Panx-1 and P2X7 at mRNA and protein levels was increased in MSU-treated HK-2 cells, which subsequently decreased upon the BA treatment. Further experiments showed that BA could combine NLRP3 inflammasome and GSDMD, destabilizing GSDMD protein. Moreover, BA protected the cell membrane from MSU-induced damage, as evidenced by Hoechst 33342 and PI double staining, lactate dehydrogenase (LDH) assays, and electron microscopy observations. These results suggest that BA is involved in the regulating Panx-1/P2X7 pathways and thus inhibits pyroptosis, highlighting its potential therapeutic effect for HN.

Bear Bile Powder Ameliorates LPS-Induced Acute Lung Injury by Inhibiting CD14 Pathway and Improving Intestinal Flora: Exploration of "Fei (Lung)-Dachang (Large Intestine) Interaction".

OBJECTIVE: To explore the effect of bear bile powder (BBP) on acute lung injury (ALI) and the underlying mechanism. METHODS: The chemical constituents of BBP were analyzed by ultra-high-pressure liquid chromatography-mass spectrometry (UPLC-MS). After 7 days of adaptive feeding, 50 mice were randomly divided into 5 groups by a random number table (n=10): normal control (NC), lipopolysaccharide (LPS), dexamethasone (Dex), low-, and high-dose BBP groups. The dosing cycle was 9 days. On the 12th and 14th days, 20 µL of Staphylococcus aureus solution (bacterial concentration of 1 × 10-7 CFU/mL) was given by nasal drip after 1 h of intragastric administration, and the mice in the NC group was given the same dose of phosphated buffered saline (PBS) solution. On the 16th day, after 1 h intragastric administration, 100 µL of LPS solution (1 mg/mL) was given by tracheal intubation, and the same dose of PBS solution was given to the NC group. Lung tissue was obtained to measure the myeloperoxidase (MPO) activity, the lung wet/dry weight ratio and expressions of CD14 and other related proteins. The lower lobe of the right lung was obtained for pathological examination. The concentrations of inflammatory cytokines including interleukin (IL)-6, tumour necrosis factor α (TNF-α ) and IL-1ß in the bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay, and the number of neutrophils was counted. The colonic contents of the mice were analyzed by 16 sRNA technique and the contents of short-chain fatty acids (SCFAs) were measured by gas chromatograph-mass spectrometer (GC-MS). RESULTS: UPLC-MS revealed that the chemical components of BBP samples were mainly tauroursodeoxycholic acid and taurochenodeoxycholic acid sodium salt. BBP reduced the activity of MPO, concentrations of inflammatory cytokines, and inhibited the expression of CD14 protein, thus suppressing the activation of NF-κB pathway (P<0.05). The lung histopathological results indicated that BBP significantly reduced the degree of neutrophil infiltration, cell shedding, necrosis, and alveolar cavity depression. Moreover, BBP effectively regulated the composition of the intestinal microflora and increased the production of SCFAs, which contributed to its treatment effect (P<0.05). CONCLUSIONS: BBP alleviates lung injury in ALI mouse through inhibiting activation of NF-κB pathway and decreasing expression of CD14 protein. BBP may promote recovery of ALI by improving the structure of intestinal flora and enhancing metabolic function of intestinal flora.

Comparison of Herbal Medicines Used for Women's Menstruation Diseases in Different Areas of the World.

Aims: This review aims to compare the use of herbal medicine used to treat women's menstruation and the prevalence of menstrual diseases in different regions, which reveal the use of herbal medicine globally and provide scientific guidance for improving women's health. Materials and Methods: The information available on herbal medicines for women between the years 2000 and 2021 was systematically collected via the library and electronic search systems such as Google Scholar, PubMed, ScienceDirect, and Web of Science as well as secondary resources including books and conference proceedings. Results: Totally, 571 ethnic medicines commonly used for women's menstruation health in Asia, Europe, Oceania, Africa, and America were accounted. Zingiber officinale Roscoe (Ginger), Ruta graveolens L. (Common rue), Angelica sinensis (Oliv.) Diels (Angelica sinensis), Foeniculum vulgare Mill (Fennel), Catharanthus roseus (L.) G. Don (Catharanthus roseus) and other medicines which have obvious advantages and long-term usage are utilized in the treatment of menstrual diseases. Family Asteraceae, Lamiaceae, Apiaceae, Fabaceae, and Zingiberaceae are the most common medicinal plant families used for such treatments. In many instances, the application of fresh parts of plants was observed because of the healers' belief regarding the higher efficiency of the medicine made from fresh plants. Edible plants are used in a wide range of countries. Conclusion: Women's menstruation health is directly related to their health condition. Traditional medicines of most ethnic groups have contributed to women's health care and treatment of gynecological diseases. Practitioners in this field have gained elaborate experience in treatments and medication, and assembled a large number of effective drugs and prescriptions. These experiences have also been inherited and developed by modern clinical application and scientific research. However, the basic research on these drugs is not sufficient, the knowledge of drug use has not been fully popularized, the advantages of drugs have not been fully utilized, and the guiding potential to modern drug research continues to be insufficient. As such, it is necessary to further promote and make a significant contribution to women's health.

Down-regulation of LGR6 promotes bone fracture recovery using bone marrow stromal cells.

OBJECTIVE: The Leucine-rich repeat-containing G-protein coupled receptor 6 (LGR6) is a well-known marker of stem cells. In present study, we aimed to further explore the effects of LGR6 on promoting osteogenic differentiation of bone marrow stromal cells (BMSCs) and bone healing. METHODS: Flow cytometry assay was used to determine the expression of BMSCs surface markers, and western blot was performed to detect the LGR6 protein expression. The osteogenic differentiation of BMSCs was qualified using ALP and ARS staining. Protein expression of osteogenic genes (ALP, Collagen I, Runx2 and OCN) were evaluated using western blot. In vivo, BMSCs transfected with sh-LGR6 or LGR6 cDNA were injected into the fracture site to establish rat fracture healing model. X-ray system and hematoxylin-eosin (HE) staining were conducted to observe the fracture recovery. Biomechanical test was performed to detect the changes of maximum load, elastic modules and bone mineral density. RESULTS: In BMSCS, CD90 and CD44 were positively expressed, while CD11b was negatively expressed. Expression level of LGR6 gradually decreased with the osteogenic differentiation of BMSCs. The osteogenic genes expression level during the osteogenic differentiation significantly increased with the down-regulation of LGR6. In vivo, 8 weeks after injection, rats treated with LGR6 knocked-down BMSCs showed increased number of fibroblasts. Maximum load, elastic modulus and the bone mineral density were enhanced with the knocking-down of LGR6. CONCLUSION: Inhibition of LGR6 promoted the osteogenic differentiation of BMSCs in vitro. Moreover, transplantation of LGR6-knockout BMSCs in rat models contributes to a better recovery after the fracture.

Development and blood compatibility assessment of electrospun polyvinyl alcohol blended with metallocene polyethylene and plectranthus amboinicus (PVA/mPE/PA) for bone tissue engineering.

INTRODUCTION: Currently, the design of extracellular matrix (ECM) with nanoscale properties in bone tissue engineering is challenging. For bone tissue engineering, the ECM must have certain properties such as being nontoxic, highly porous, and should not cause foreign body reactions. MATERIALS AND METHODS: In this study, the hybrid scaffold based on polyvinyl alcohol (PVA) blended with metallocene polyethylene (mPE) and plectranthus amboinicus (PA) was fabricated for bone tissue engineering via electrospinning. The fabricated hybrid nanocomposites were characterized by scanning electron microscopy (SEM), Fourier transform and infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), contact angle measurement, and atomic force microscopy (AFM). Furthermore, activated partial thromboplastin time (APTT), prothrombin time (PT), and hemolytic assays were used to investigate the blood compatibility of the prepared hybrid nanocomposites. RESULTS: The prepared hybrid nanocomposites showed reduced fiber diameter (238±45 nm) and also increased porosity (87%) with decreased pore diameter (340±86 nm) compared with pure PVA. The interactions between PVA, mPE, and PA were identified by the formation of the additional peaks as revealed in FTIR. Furthermore, the prepared hybrid nanocomposites showed a decreased contact angle of 51°±1.32° indicating a hydrophilic nature and exhibited lower thermal stability compared to pristine PVA. Moreover, the mechanical results revealed that the electrospun scaffold showed an improved tensile strength of 3.55±0.29 MPa compared with the pristine PVA (1.8±0.52 MPa). The prepared hybrid nanocomposites showed delayed blood clotting as noted in APTT and PT assays indicating better blood compatibility. Moreover, the hemolysis assay revealed that the hybrid nanocomposites exhibited a low hemolytic index of 0.6% compared with pure PVA, which was 1.6% suggesting the safety of the developed nanocomposite to red blood cells (RBCs). CONCLUSION: The prepared nanocomposites exhibited better physico-chemical properties, sufficient porosity, mechanical strength, and blood compatibility, which favors it as a valuable candidate in bone tissue engineering for repairing the bone defects.

The prognostic value of combined NT-pro-BNP levels and NIHSS scores in patients with acute ischemic stroke.

OBJECTIVE: Determining the prognoses of patients with acute ischemic stroke is difficult. Therefore, the aim of this study was to evaluate whether the combined assessment of plasma N-terminal pro-brain natriuretic peptide (NT-pro-BNP) and the National Institutes of Health Stroke Scale (NIHSS) variables is relevant to the prognosis of patients with acute cerebral ischemic infarction in-hospital. METHODS: We enrolled 122 patients who were within three days of onset of acute ischemic stroke. We measured the plasma NT-pro-BNP level of each patient within 72 hours and recorded the NIHSS score on admission. The factors associated with death were investigated using a multivariate logistic regression analysis. RESULTS: Twenty-three patients (18.85%) died during hospitalization. The frequency of atrial fibrillation (AF), the NIHSS score on admission (8.69±4.87 in the survival group vs. 14.48±2.54 in the deceased group, p<0.001) and the plasma NT-pro-BNP level (median: 926.30 pg/mL in the survival group vs. 3,280 pg/mL in the deceased group, p<0.001; Lg NT-pro-BNP 2.82±0.66 in the survival group vs. 3.46±0.52 in the deceased group, p<0.001) were each significantly higher in the deceased group than in the survival group. The optimal cut-off levels for the NT-pro-BNP level and NIHSS score to distinguish the deceased group from the survival group were 1,583.50 pg/mL and 12.5, respectively. Patients with both elevated NT-pro-BNP levels (>1,583.50 pg/mL) and NIHSS scores on admission (NIHSS >12.5) had a substantially higher mortality rate than those without elevated NT-pro-BNP levels and NIHSS scores (89.47% vs. 9.84%, p<0.001). A multivariate logistic regression analysis demonstrated that a NT-pro-BNP level >1,583.50 pg/mL (OR, 5.001; 95% CI, 1.233 to 20.287, p=0.024) and a NIHSS score >12.5 (OR, 1.465; 95% CI, 1.191 to 1.801, p<0.001) were each independent factors associated with in-hospital death. CONCLUSION: The plasma NT-pro-BNP level and the NIHSS score added independent and incremental contributions to the prognostic stratification of patients with acute ischemic stroke.