Hair Follicles as a Critical Model for Monitoring the Circadian Clock.

Clock (circadian) genes are heterogeneously expressed in hair follicles (HFs). The genes can be modulated by both the central circadian system and some extrinsic factors, such as light and thyroid hormones. These circadian genes participate in the regulation of several physiological processes of HFs, including hair growth and pigmentation. On the other hand, because peripheral circadian genes are synchronized with the central clock, HFs could provide a noninvasive and practical method for monitoring and evaluating multiple circadian-rhythm-related conditions and disorders among humans, including day and night shifts, sleep-wake disorders, physical activities, energy metabolism, and aging. However, due to the complexity of circadian biology, understanding how intrinsic oscillation operates using peripheral tissues only may be insufficient. Combining HF sampling with multidimensional assays such as detection of body temperature, blood samples, or certain validated questionnaires may be helpful in improving HF applications. Thus, HFs can serve as a critical model for monitoring the circadian clock and can help provide an understanding of the potential mechanisms of circadian-rhythm-related conditions; furthermore, chronotherapy could support personalized treatment scheduling based on the gene expression profile expressed in HFs.

Exploring and validating the metastasis mechanism of pathenolide interfering with cutaneous melanoma through ER stress-dependent apoptosis based on the network pharmacology.

BACKGROUND: This study aimed to explore the mechanism of parthenolide in inhibiting melanoma metastasis through network pharmacology and cell experiment. MATERIALS AND METHODS: This research obtained the targets of the drug from the HERB database and PubChem database, the differential expression gene of metastatic cutaneous melanoma was obtained by differentially expression gene analysis of four Gene Expression Omnibus (GEO) datasets. The intersection of drug targets and differentially expression genes were considered to be related to drugs that inhibit metastasis of cutaneous melanoma. The STRING database was used to construct the protein-protein interaction (PPI) network, and cytohubba package in Cytoscape software was used to rank the PPI network targets. The enrichment analysis was used to screen out the relevance Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and gene ontology to explain the underlying mechanism of drug inhibiting the cutaneous melanoma metastatic; cell viability, apoptosis, cell migration and protein levels were assessed using cell counting kit-8 (CCK-8) assay, Annexin V-FITC/PI assay, wound healing assay, respectively. Finally, combining pathway maps and literature, we detected ATF4 and proteins upstream and downstream of ATF4 through Western blot. RESULTS: A total of 87 targets were screened out from the drug databases, and a total of 1635 differentially expression genes was obtained from the differentially expression genes analysis of GEO datasets, a total of nine targets (VEGFA, ANXA5, ICAM1, SELE, NFKBIA, ATF4, CTNNB1, SELP and HPGDS) were considered to be related to drugs that inhibit metastasis of cutaneous melanoma. The result of enrichment analysis showed that the drug inhibits the metastatic of cutaneous melanoma through multiple pathways such as TNF signalling pathway, lipid and atherosclerosis and fluid shear stress and atherosclerosis, relevance multiple biological processes, cellular components and molecular function; cell experiments showed that parthenolide could inhibit tumour cell migration and induce a decrease of cell viability. Flow cytometry results showed that parthenolide induced tumour cell apoptosis. Western blot results suggested that parthenolide exerted therapeutic effects by regulating ATF4 protein and its upstream and downstream proteins, namely endoplasmic reticulum (ER) stress signalling pathway. CONCLUSION: Parthenolide induces ER stress-dependent apoptosis in melanoma cells.

Immunogenic exosome-encapsulated black phosphorus nanoparticles as an effective anticancer photo-nanovaccine.

Tumor vaccines are a promising form of cancer immunotherapy, but difficulties such as neo-antigen identification, activation of immune cells, and tumor infiltration prevent their clinical breakthrough. Interestingly, nanotechnology-based photothermal therapy (PTT) has great potential to overcome these barriers. Previous studies have shown that serum exosomes (hEX) from hyperthermia-treated tumor-bearing mice displayed an array of patient-specific tumor-associated antigens (TAAs), and strong immunoregulatory abilities in promoting dendritic cell (DC) differentiation and maturation. Here, we developed a tumor vaccine (hEX@BP) by encapsulating black phosphorus quantum dots (BPQDs) with exosomes (hEX) against a murine subcutaneous lung cancer model. In comparison with BPQDs alone (BP), hEX@BP demonstrated better long-term PTT performance, greater elevation of tumor temperature and tumor targeting efficacy in vivo. Vaccination with hEX@BP in combination with PTT further demonstrated an outstanding therapeutic efficacy against established lung cancer, and promoted the infiltration of T lymphocytes into the tumor tissue. Our findings demonstrated that hEX@BP might be an innovative cancer photo-nanovaccine that offers effective immuno-PTT against cancers.

Antioxidative and Immunomodulatory Activities of the Exopolysaccharides from Submerged Culture of Hen of the Woods or Maitake Culinary-Medicinal Mushroom, Grifola frondosa (Agaricomycetes) by Addition of Rhizoma gastrodiae Extract and Its Main Components.

Grifola frondosa (hen of the woods or maitake) is a famous culinary-medicinal mushroom, and its exopolysaccharides (EPSs) have biological activities with or without supplementation with exogenous additives. In this study, a Rhizoma gastrodiae extract was added to a G. frondosa fermentation system. P-hydroxylbenzaldehyde (HBA), the main product of R. gastrodiae, had the highest utilization rate in the fermentation process (42%). In addition, the EPSs of G. frondosa after addition of R. gastrodiae extract (REPS), of HBA (HEPS), or of a standard solution according to the main component ratio of R. gastrodiae extract (CEPS) were obtained. We then determined the antioxidant and immunomodulatory activities of EPS, REPS, HEPS, and CEPS. Overall, REPS showed the highest antioxidant activities compared with EPS and HEPS (P < 0.05) but similar to that of CEPS (P > 0.05). The half-inhibitory concentration (ED50) values of REPS (< 4 mg/mL) were lower than those of EPS, HEPS, and CEPS. Moreover, REPS was better able to stimulate phagocytosis and nitric oxide production of RAW 264.7 macrophages than were the others, without a significant difference from CEPS (P > 0.05). An interesting and important finding is that a R. gastrodiae extract can increase antioxidant and immunomodulatory activities of EPS preparations from G. frondosa, and the standard solution of the main components of the R. gastrodiae extract may be better for simulating fermentation performed by G. frondosa and biological activities of its major products.

Comparative analysis of robotic vs laparoscopic radical hysterectomy for cervical cancer.

BACKGROUND: Cervical cancer is the most common gynecological malignancy, ranking first in female reproductive malignancies with more than 500000 new cases and 275000 deaths each year. Traditionally, open radical hysterectomy is considered the standard surgical procedure for the treatment of resectable cervical cancer. The latest guidelines from the National Comprehensive Cancer Network and the European Society of Gynecological Oncology suggest that open surgery and laparoscopic surgery (using traditional laparoscopic or robotic techniques) are the main surgical approaches for radical hysterectomy for patients with stage IA2-IIA cervical cancer. Robotic surgery has been increasingly used in abdominal surgery and has shown more beneficial effects. AIM: To analyse the perioperative conditions, complications, and short-term and long-term effects in patients undergoing robotic radical hysterectomy (RRH) and laparoscopic radical hysterectomy (LRH) to compare their clinical efficacy, safety, and feasibility. METHODS: The perioperative data of patients undergoing RRH and LRH were extracted and collected from the database of surgical treatments for cervical cancer for statistical analysis. RESULTS: Of the patients, 342 underwent LRH for cervical cancer, and 216 underwent RRH. The total complication rate was 9.65% (20 patients) in the RRH group and 17.59% (60 patients) in the LRH group. The complication rate was significantly lower in the RRH group than in the LRH group. There was no significant difference in the follow-up period (P = 0.658). The total recurrence rates were 15.7% and 12% in the RRH and LRH groups, respectively. The progression-free survival time was 28.91 ± 15.68 mo and 28.34 ± 15.13 mo in the RRH and LRH groups, respectively (P = 0.669). The overall survival (OS) rates were 92.13% and 94.45% in the RRH and LRH groups, respectively (P = 0.292). The OS time was 29.87 ± 15.92 mo and 29.41 ± 15.14 mo in the RRH and LRH groups, respectively (P = 0.732). The survival curves and the progression-free survival curves were not statistically significantly different between the two groups (P = 0.407 and 0.28, respectively). CONCLUSION: RRH is associated with significantly less operative time and blood loss than LRH. The two procedures have similar complication rates, OS, and progression-free survival time.

PSORI-CM02 alleviates IMQ-induced mouse dermatitis via differentially regulating pro- and anti-inflammatory cytokines targeting of Th2 specific transcript factor GATA3.

The IL-17-producing CD4+ T cell and γδT cells play critical roles in the pathogenesis of psoriasis (PS). PSORI-CM02 is a representative herbal formula for the treatment for PS in South China. It was confirmed to improve PS without obvious side effects in the clinic. Here we sought to clarify whether and how PSORI-CM02 regulates T cell differentiation and functions in IMQ-induced psoriasis-like BALB/c mouse model. Mice pre-treated 3 days with PSORI-CM02 significantly alleviated skin inflammation, as reduced in PASI score and classic psoriatic characteristics in pathological sections. CD3 and CD4 positive T cells were also fewer in the skin lesions of PSORI-CM02 groups, comparing to control group. PSORI-CM02 also decreased pro-inflammatory IFNγ mRNA and IL-17 A mRNA, while increased IL-4 mRNA in mouse skin lesions. In skin draining lymph nodes (DLN), PSORI-CM02 reduced the ratio of γδT cells and inhibited their function of producing IL-17 A. Nevertheless PSORI-CM02 had no effects on the ratio of total TCRß+T cells and CD4 + T cells. But it regulated CD4 + T helper cells differentiation, and resulted in the decreasing percentage of IFNγ producing Th1 cells and IL-17 A producing Th17 cells, while increasing the ratio of IL-4 producing Th2 cells in DLN. Further data showed that PSORI-CM02 promote expression of Th2 specific transcript factor GATA3, but had no effects on T-bet and RORγ. Thus, we tentatively interpret that PSORI-CM02 impairs IMQ-induced psoriasis by promoting Th2 cell response targeting of GATA3.

Enhanced Neurogenesis and Collaterogenesis by Sodium Danshensu Treatment After Focal Cerebral Ischemia in Mice.

Ischemic stroke remains a serious threat to human life. There are limited effective therapies for the treatment of stroke. We have previously demonstrated that angiogenesis and neurogenesis in the brain play an important role in functional recovery following ischemic stroke. Recent studies indicate that increased arteriogenesis and collateral circulation are determining factors for restoring reperfusion and outcomes of stroke patients. Danshensu, the Salvia miltiorrhiza root extract, is used in treatments of various human ischemic events in traditional Chinese medicine. Its therapeutic mechanism, however, is not well clarified. Due to its proposed effect on angiogenesis and arteriogenesis, we hypothesized that danshensu could benefit stroke recovery through stimulating neurogenesis and collaterogenesis in the post-ischemia brain. Focal ischemic stroke targeting the right sensorimotor cortex was induced in wild-type C57BL6 mice and transgenic mice expressing green fluorescent protein (GFP) to label smooth muscle cells of brain arteries. Sodium danshensu (SDS, 700 mg/kg) was administered intraperitoneally (i.p.) 10 min after stroke and once daily until animals were sacrificed. To label proliferating cells, 5-bromo-2'-deoxyuridine (BrdU; 50 mg/kg, i.p.) was administered, starting on day 3 after ischemia and continued once daily until sacrifice. At 14 days after stroke, SDS significantly increased the expression of vascular endothelial growth factor (VEGF), stromal-derived factor-1 (SDF-1), brain-derived neurotrophic factor (BDNF), and endothelial nitric oxide synthase (eNOS) in the peri-infarct region. SDS-treated animals showed increased number of doublecortin (DCX)-positive cells. Greater numbers of proliferating endothelial cells and smooth muscle cells were detected in SDS-treated mice 21 days after stroke in comparison with vehicle controls. The number of newly formed neurons labeled by NeuN and BrdU antibodies increased in SDS-treated mice 28 days after stroke. SDS significantly increased the newly formed arteries and the diameter of collateral arteries, leading to enhanced local cerebral blood flow recovery after stroke. These results suggest that systemic sodium danshensu treatment shows significant regenerative effects in the post-ischemic brain, which may benefit long-term functional recovery from ischemic stroke.

Omnipotent phosphorene: a next-generation, two-dimensional nanoplatform for multidisciplinary biomedical applications.

Phosphorene, also known as single- or few-layer black phosphorus (FLBP), is a new member of the two-dimensional (2D) material family and has attracted significant attention in recent years for applications in optoelectronics, energy storage and biomedicine due to its unique physicochemical properties and excellent biocompatibility. FLBP is regarded as a potential biological imaging agent for cancer diagnosis due to its intrinsic fluorescence (FL) and photoacoustic (PA) properties and negligible cytotoxicity. FLBP-based photothermal and photodynamic therapies have emerged with excellent anti-tumour therapeutic efficacies due to their unique physical properties, such as near-infrared (NIR) optical absorbance, large extinction coefficients, biodegradability and reactive oxygen species (ROS) or heat generation upon light irradiation. Furthermore, FLBP is a promising drug delivery platform because of its high drug-loading capacity due to its puckered layer structure with an ultralarge surface area, and FLBP is size-controllable with facile surface chemical modification. Because of the marked advantages of FLBP nanomaterials in biomedical applications, an overview of the latest progress and paradigms of FLBP-based nanoplatforms for multidisciplinary biomedical applications is presented in this tutorial review.

[Determination of Total Sulfur Dioxide in Chinese Herbal Medicines via Triple Quadrupole Inductively Coupled Plasma Mass Spectrometry].

As an important treatment method, sulfur fumigation plays an essential role in the production and preservation of traditional Chinese herbal medicines. Although there is strict regulation on the use of sulfur dioxide, the abuse of sulfur dioxide still occurred from time to time. And the public faces a high risk of exposure. Because of the poor precision and tedious preparation procedures of traditional recommended titration, the accurate and convenient determination of sulfur dioxide in Chinese herbal medicines is still a critical analytical task for medicines safety and the public health. In this study, an accurate, high-throughput, and convenient method for the absolute determination of SO2 in Chinese herbal medicines based on triple quadrupole inductively coupled plasma mass spectrometry (ICP-MS/MS) technique is developed. The study compared the quantitative ability for sulfur when the ICP-MS operated under traditional single quadrupole (QMS) mode and novel triple quadrupole (MS/MS) mode with three Reaction/Collision cell condition (no gas, helium, and oxygen). The result indicated that when the concentration of sulfate ranging from 0.5 to 100 mg · L⁻¹, isotopic ³4S can be selected as quantitative ion either the ICP-MS operated under the QMS mode or MS/MS mode. The use of helium in the Reaction/Collision cell decreased the single intensity of background ions. Better than QMS mode, the MS/MS mode can effectively reduced background interference. But there are no significant differences about the linear range and limit of detection. However, when the ICP-MS operated under MS/MS mode and oxygen was used as reaction gas in the Reaction/Collision cell, the ICP-MS/MS provided an interference-free performance, the linear range and limit of detection improved significantly. Either ³²S or ³4S exhibits an excellent linearity (r > 0.999) over the concentration range of 0.02-100 mg · L⁻¹, with a limit of detection of 5.48 and 9.76 µg · L⁻¹ for ³²S¹6O4²â» and ³4S¹6O4²â», respectively. The Chinese herbal medicines was treated using microwave digestion added 6 mL nitric acid and 2 mL hydrogen peroxide before analysis. The amount of nitric acid and hydrogen peroxide were optimized. The method was validated using Chinese herbal standard reference material GBW10020. The sample was treated and detected in six parallel, and the average concentrations obtained using the developed method (0.42% ± 0.01%) is in excellent agreement with the standard concentration (0.41%± 0.03%). The study demonstrates an accurate and convenient approach for the quantification of SO2 in Chinese herbal medicines.

[Optimization of Extraction Technology of Yiyi Fuzi Baijiang Powder by Orthogonal Design].

Objective: To optimize the optimum extraction technology of Yiyi Fuzi Baijiang powder. Methods: Orthogonal design was used,the extraction rate of Coicis Semen oil was used to determine the ethanol extraction technology of Coicis Semen,the extraction rate of protocatechuic acid,chlorogenic acid and benzoylmesaconine were used to determine the water extraction technology of Aconiti Lateralis Radix Praeparata and Herba Patriniae. Results: The optimum extraction technology of Coicis Semen was obtained,alcohol was consumed as four times of the herb amount,and ultrasonic extracted three times with 20 minutes each time; the optimum extraction technology of Aconiti Lateralis Radix Praeparata and Herba Patriniae was obtained,water was consumed as ten times of the herb amount,and extracted for 1. 0 h for Aconiti Lateralis Radix Praeparata and then Herba Patriniae was added and extracted for 45 minutes for the first time; in the second time,water was consumed as 8 times of the herb amount,and extracted for 30 minutes for dregs of herbs. Conclusion: The optimum extraction technology is stable and feasible,in which can give a reference to the preparation research of Yiyi Fuzi Baijiang powder.

A new biflavone glucoside from the roots of Stellera chamaejasme.

The present study investigated the chemical constituents of the roots of Stellera chamaejasme (Thymelaeaceae). One new biflavone glucoside (1), along with other thirteen known compounds (2-14), was isolated by repeated column chromatographic methods and their structures were elucidated on the basis of spectral analyses. The cytotoxic activities of selected compounds were evaluated against four human cancer cell lines (A549, BEL-7402, HCT-116, and MDA-MB-231) by the SRB assay method. Compound 9 showed remarkable cytotoxicity against BEL-7402 with IC50 value being 0.65 µg·mL(-1); compounds 7, 8, and 12 exhibited significant cytotoxic activity against A549 with IC50 values being 2.38, 1.57, and 2.35 µg·mL(-1), respectively.

Effect of alternate-day fasting therapy combined with Linggui Zhugan Decoction () on hepatic oxidative stress in hyperlipidemic rat.

OBJECTIVE: To observe the effect of alternate-day fasting (ADF) therapy combined with Linggui Zhugan Decoction (, LZD) on hepatic oxidative stress and blood lipids in hyperlipidemic rats. METHODS: Fifty-two Wistar rats were randomly assigned to two groups: the high-fat-diet (HF) group and the normal-diet (ND) group. Hyperlipidemia was induced by feeding rats with high-cholesterol-diet for 5 weeks. Then the HF group was randomized to the model-control (MC) group, the alternate-day-fasting (ADF) group, and the ADF combined with LZD (AL) group. The ND group was regarded as the negative control (NC) group. The AL and ADF groups were put on fast for 24 h on alternate days for 4 weeks. The AL group was administrated with LZD on the fast days. Body weight and food intake were measured once a week. After 4-week ADF, blood sample was collected for determination of plasma total cholesterol (TC), triglyceride (TG), low-density-lipoprotein cholesterol (LDL-C) and high-density-lipoprotein cholesterol (HDL-C). Liver oxidative stress parameters including total superoxide dismutase (T-SOD) activity, malondialdehyde (MDA) level and glutathione (GSH) content were also tested. RESULTS: Body weight in the HF group decreased significantly (P<0.01). TC, TG, HDL-C, and LDL-C concentrations in the HF group were higher than those in the NC group (P<0.01), respectively. T-SOD in the HF group was clearly lower than that in the NC group (P<0.05). After 4-week intervention, body weight, TC and TG concentrations in the ADF and AL groups declined significantly, respectively, compared to MC group (P<0.05). GSH in the ADF and AL groups were much higher than those in the MC group (P<0.01). MDA level was also greatly higher in the ADF group as compared with the NC group (P<0.05). CONCLUSION: ADF therapy combined with LZD may be used as an effective combination approach for treatment of hyperlipidemia and oxidative stress damage.

Casticin induces breast cancer cell apoptosis by inhibiting the expression of forkhead box protein M1.

Casticin is an active ingredient derived from Fructus Viticis, a traditional Chinese medicine. This study aimed to investigate the role of forkhead box O3 (FOXO3a) in breast cancer cells and examine the regulatory mechanisms of FOXO3a in response to casticin treatment of the cells by ELISA, flow cytometry, small interfering RNA (siRNA) transfection and western blot analysis. Casticin treatment induced apoptosis and reduced the expression of the transcription factor forkhead box protein M1 (FOXM1). In addition, FOXM1 repression induced by casticin treatment was associated with the activation of FOXO3a via increased dephosphorylation. Notably, silencing FOXO3a expression by siRNA-mediated gene knockdown attenuated casticin-mediated apoptosis. Collectively, these findings suggest that FOXO3a is a critical mediator of the inhibitory effects of casticin on apoptosis in breast cancer cells.

Baicalein decreases side population proportion via inhibition of ABCG2 in multiple myeloma cell line RPMI 8226 in vitro.

OBJECTIVE: To investigate the effect of baicalein on side population in human multiple myeloma cell line RPMI 8226 and the underlying molecular mechanisms in vitro and in silico. METHODS: MTT assay was applied to detect the anti-proliferation effect of baicalein. The detection of side population cells is based on the Hoechst 33342 exclusion assay technique and flow cytometric analysis. Western blotting assay was used to explore the expression of ABCG2 protein. Homology modeling and molecular docking were performed with Discovery Studio 2.1. RESULTS: Baicalein decreased both cell viability with IC50=168.5 µM and the proportion of SP cells in a dose-dependent manner. Correspondingly, it significantly decreased the expression level of ABCG2 protein. Baicalein also shared similar binding sites and modes with fumitremorgin C to the protein. CONCLUSIONS: Baicalein possessed novel anticancer properties, such as anti-proliferation and drug efflux inhibition in side population cells, which suggested its potential feature of targeting cancer stem cells of multiple myeloma.

[Application of grading evaluation on facial nerve function of Bell's palsy treated with electroacupuncture].

OBJECTIVE: To explore the correlation of facial nerve injury degree with facial contraction degree induced by electric stimulation in the treatment of Bell's palsy with electroacupuncture, and the significance in elec tric reaction grading evaluation. METHODS: Sixty-eight cases of Bell's palsy were enrolled. The positive and negative electrodes of the acupuncture treatment apparatus were attached to the needle handles at the 3 groups of points, named Taiyang (EX-HN 5)-Yangbai(GB 14), Xiaguan (ST 7)-Quanliao (SI 18) and Heliao (LI 19)-Jiachengjiang (Extra). The disperse-dense wave was applied. According to the severity of local muscle contraction after needling, the electric reaction was divided into 4 grades, named superior, moderate, poor and no reaction. After acupuncture and electroacupuncture, the efficacy was evaluated in accordance with the different electric reaction grades. METHODS: The curative rate was 100.0% (44/44) in patients with superior electric reaction, was 100.0% (7/7) in patients with moderate electric reaction, was 18.2% (2/11) in patients with poor electric reaction and was 0 (0/6) in patients with noelectric reaction. The difference was significant statistically in comparison of 4 groups (P<0.01). The superiority correlation presented between the efficacy and electric reaction grade (P< 0.001). The higher the superiority of electric reaction grade was, the better the efficacy was. The difference in the efficacy among different electric reaction grades was significant statistically (P<0.001). And the course of treatment was the shortest for those with the high superiority of electric reaction. CONCLUSION: The reaction grade of electric stimulation is conform to the facial nerve injury grading in Bell's palsy. The contraction degree of facial mimetic muscle induced by electroacupuncture stimulation is closely correlated with severity of disease. Based on the electric reaction, the facial nerve injury severity can be understood generally and the prognosis be judged.

Four new diterpenoids from the roots of Euphorbia fischeriana.

Four new diterpenoids (1,4,5,9), together with 7 known diterpenoids (2,3,6-8,10,11), were isolated from the roots to Euphorbia fischeriana. On the basis of 1D and 2D NMR, HR-ESI-MS spectroscopic analysis, structures of the new compounds were elucidated as 11ß-hydroxy-8,14-epoxy-ent-abieta-13(15)-en-16,12-olide (1), 3,20-dihydroxy-ent-1(10), 15-rosadiene (4), 3,7-dihydroxy-ent-1(10), 15-rosadiene (5), ingenol 6,7-epoxy-3- tetradecanoate (9). The compounds isolated were evaluated for their cytotoxicity against four cancer cell lines (A549, BEL7402, HCT116, and MDA-MB-231). Three ingenol diterpenoids (9-11) showed significant cytotoxicity against A549 with IC50 value of 3.35, 2.85, 2.88 µg/mL, respectively.

Scutellaria extract decreases the proportion of side population cells in a myeloma cell line by down-regulating the expression of ABCG2 protein.

BACKGROUND AND AIMS: Scutellaria is one of the most popular traditional Chinese herbal remedies against various human diseases, including cancer. In this study, we examined the active effects of Scutellaria extract and its main flavonoid constituents on the proportion of side population cells within human multiple myeloma cell line RPMI8226 in vitro and explored the potential molecular mechanisms involved. MATERIALS AND METHODS: The contents of flavonoids in ethanolic extract of Scutellaria baicalensis Georgi were determined using high performance liquid chromatography. The antiproliferative effect of the ethanolic extract on RPMI-8226 was determined by CCK assay. Apoptosis was measured by annexin combining with propidium iodide in a flow cytometer. Cell cycle analysis was performed by propidium iodide staining in combination with flow cytometry analysis. Hoechst 33342 exclusion assay was used for the identification of side population within RPMI8226 cells. The expression of ABCG2 protein was assessed by Western blotting assay. RESULTS: The content of major flavonoids constitutents of Scutellaria extract was baicalin (10.2%), wogonoside (2.50%), baicalein (2.29%), and wogonin (0.99%), respectively. The crude Scutellaria extract did not show significant anti-proliferative effect, apoptosis induction and cell cycle arrest in RPMI-8226 within the concentrations of 1-75µg/mL. However, the ethanolic extract, baicalein, wogonin and baicalin reduced the side population cells in RPMI-8226, and data showed that baicalein and wogonin had stronger inhibitory effects. Correspondingly, they also exhibited significant effects on decreasing the expression level of ABCG2 protein in RPMI-8226 in vitro. CONCLUSIONS: Our results for the first time demonstrated a novel mechanism of action for Scutellaria extract and its main active flavonoids, namely targeting SP cells by modulating the expression of ABCG2 protein. This study provides an insight for new therapeutic strategies targeting cancer stem cells of multiple myeloma.

Age differences in clinical characteristics, health care, and outcomes after ischemic stroke in China.

BACKGROUND AND PURPOSE: Little information is available on the effects of age on health care and outcomes of ischemic stroke (IS) in China. Our aim was to evaluated risk factors, health care, and outcomes among age groups including ≤ 45, 46-65, 66-79, and ≥ 80 years and to find whether the outcome was affected by age and health care. METHODS: CNSR is a nationwide prospective registry for patients admitted with acute stroke and prospectively followed up 12-month outcomes. Demographics, socioeconomics, risk factors, health care, and outcomes were analyzed among age groups, and multivariate regression analysis was used to determine the association of outcome and age and health care. RESULTS: We identified 12,415 acute IS patients for analysis. Of 1179 (9.50%) were aged ≥ 80 years. In terms of risk factors, cardiac diseases were significantly more frequent in patients ≥ 80 years, behavioral risk factors were more common in younger patients, and hypertension, hyperlipidemia, and diabetes were more seen in 46-79 patients. The use of health care varied among groups and was significantly lower in ≥ 80 years especially in secondary prevention. The very old patients had the worst outcomes even after adjusting by prognostic factors; however, adjusting forward by health care, the extent of differences decreased. CONCLUSIONS: In CNSR, differences in stroke clinic characteristics and health care were observed among various age groups, and the old patients, receiving lower levels of stroke care, had the worst outcomes. Knowledge of the age differences in ischemic stroke may be helpful to appropriately allocate the limited health resources and to improve stroke outcomes.

Antithrombotic management of ischaemic stroke and transient ischaemic attack in China: a consecutive cross-sectional survey.

1. Little is known about the prevention of secondary stroke in China. In the present study, we assessed the status of antithrombotic management of stroke patients in clinics across China. 2. A cross-sectional survey was conducted in 19 urban neurological clinics. All subjects diagnosed with ischaemic stroke (IS) or transient ischaemic attack (TIA) were enrolled consecutively in the study. Face-to-face interviews were conducted by research assistants using questionnaires on the day of enrollment. The data recorded included demographic and clinical characteristics, medication and reasons for not using medication. Independent predictors for the prescription of antiplatelet drugs were determined using multivariate logistic regression models. 3. Of the 2283 patients with IS or TIA enrolled in the study (34.7% women; mean ( +/- SD) age 65.8 +/- 11.6 years), 1719 (75.3%) had a prescription for antiplatelet therapy. Of the 108 patients with atrial fibrillation, only 14 (13.0%) were receiving warfarin therapy. The main independent factors significantly associated with being on antiplatelet therapy were having basic health insurance (odds ratio (OR) 1.47; 95% confidence interval (CI) 1.09-1.99), government insurance and labour insurance (OR 1.63; 95% CI 1.03-2.59) and a monthly income of > 500 yuan (US$66.70; OR 2.14; 95% CI 1.51-3.03). Being older (OR 0.70; 95% CI 0.50-0.99) and having a severe disability (OR 0.68; 95% CI 0.49-0.97) were associated with lower odds of receiving antiplatelet therapy. 4. Based on the survey results, adherence to guidelines for antithrombotic management in neurological clinics in China is poor. The main reasons contributing to the less than optimal management of stroke patients include negative attitudes among neurologists, a lack of medical insurance, a lower income and being elderly and/or severely disabled.

The protective effects of Polygonum multiflorum stilbeneglycoside preconditioning in an ischemia/reperfusion model of HUVECs.

AIM: To investigate the protective effects of preconditioning human umbilical vein endothelial cells (HUVECs) with Polygonum multiflorum stilbeneglycoside (PMS) under anoxia/reoxygenation (A/R), and the mechanism of protection. METHODS: Prior to A/R, HUVECs were incubated with PMS (0.6 x 10(-11), 1.2 x 10(-11), or 2.4 x 10(-11) mol/L) for 3 h. Cell injury was subsequently evaluated by measuring cell viability with an MTT assay and lactate dehydrogenase (LDH) release, whereas lipid peroxidation was assayed by measuring malondialdehyde (MDA) content. Antioxidant capacity was quantified by superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Nitric oxide (NO) production was determined by nitrite accumulation. Endothelial NO synthase (eNOS) and inducible NOS (iNOS) protein expression was detected by Western blotting. Guanylate cyclase activity and cyclic GMP (cGMP) activity were assessed by an enzyme immunoassay kit. RESULTS: PMS incubation attenuated A/R-induced injury in a concentration-dependent manner, as evidenced by a decrease in LDH activity and an increase in cell viability. PMS exerted its protective effect by inhibiting the A/R-mediated elevation of MDA content, as well as by promoting the recovery of SOD and GSH-Px activities. Additionally, PMS incubation enhanced NO and cGMP formation by increasing iNOS expression and guanylate cyclase activity. The protective effects of PMS were markedly attenuated by NOS inhibitor L-NAME, soluble guanylate cyclase inhibitor ODQ or PKG inhibitor KT5823. CONCLUSION: PMS preincubation resulted in the enhancement of antioxidant activity and anti-lipid peroxidation. The NO/cGMP/cGMP-dependent protein kinase (PKG) signaling pathway was involved in the effect of PMS on HUVECs.