Network pharmacology and experimental verification of the potential mechanism of Er-Xian decoction in aplastic anemia.

To investigate the potential mechanism of Er-Xian decoction (EXD) in treating aplastic anemia (AA), the active components of EXD were screened by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the targets of the components were predicted by the Swiss Target Prediction database. AA targets were collected from the GeneCards, OMIM, DisGeNET, PharmGKB, DrugBank, and TTD databases, the intersection of AA targets and EXD targets was calculated, and an herb-component-target network was constructed by Cytoscape 3.7.2 software. The STRING database was used for protein‒protein interaction (PPI) analysis, and Cytoscape 3.7.2 software was used to construct a PPI network and perform topology analysis. The core targets were imported into the DAVID database for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. The molecular docking software AutoDock was used to measure the affinity between active components and key targets. Finally, we established a mouse model of AA and verified the key targets and signaling pathways of EXD by RT‒PCR, ELISA and Western blot analysis. A total of 53 active components were screened from EXD, 2516 AA-related targets were collected, and 195 common targets were obtained. An herb-component-target network and a PPI network were successfully constructed, and 36 core targets were selected from the PPI network. The main active components of EXD include luteolin, kaempferol, berberine, etc., and key targets include PIK3CA, AKT1, STAT3, etc. GO functional enrichment analysis showed that cell components, molecular functions and biological processes with significant correlations were macromolecular complexes, protein serine/threonine/tyrosine kinase activity and protein phosphorylation, respectively. KEGG pathway analysis showed that the pathways with significant correlations included the PI3K-Akt signaling pathway and JAK-STAT signaling pathway. Molecular docking results showed that the tested key targets had good affinity for the corresponding active components. In AA mice, we found that EXD significantly increased white blood cell count, red blood cell count, platelet count and hemoglobin levels, increased mRNA levels of PIK3CA, PIK3CD, AKT1, JAK2, STAT3 and MAPK1, and promoted phosphorylation of PI3K, AKT, ERK1/2 and STAT3. In summary, EXD acts on PI3K, AKT, STAT3 and other targets through berberine, luteolin, quercetin and other components to regulate the PI3K-Akt pathway, JAK-STAT pathway and other pathways, thus exerting its therapeutic effect on AA. This study explained the Chinese medicine theory of treating AA with EXD by tonifying kidney-yang and provides a scientific basis for the use of EXD in treating AA.

Metabolite Profiling of Talatisamine in Heart Tissue After Oral Administration and Analysis of Cardiac Bioactivities.

The lateral roots of the Aconitum carmichaelii ("Fuzi") have been used for centuries as a cardiotonic in China. The diterpenoid alkaloid talatisamine (TA) is a major bioactive component of Fuzi, but the identity and bioactivities of the TA metabolites have not been examined in detail. In this study, metabolite profiling of TA was performed in rat heart by UPLC-MS following oral administration. Metabolites were identified by comparing protonated molecules, fragmentation patterns, and chromatographic behaviors with those of standard compounds. Metabolites of TA were then prepared and tested for cardiotonic activity on isolated frog hearts. The metabolite cammaconine, a C19 diterpenoid alkaloid with a hydroxyl group at C-18, exhibited substantial cardiotonic activity during frog heart perfusion. To further investigate the structure-cardiac effect relationships, a series of C19-diterpenoid alkaloids with 18-OH were prepared. Eight tested compounds (5: -12: ) demonstrated measurable cardioactivity, of which compound 5: with an N-methyl group and compound 7: with a methoxy at C-16 showed stronger effects on ventricular contraction than the other compounds. Thus, 18-OH is a critical structural feature determining cardiotonic activity, and efficacy is improved by the presence of N-methyl or methoxy at C-16. Preliminary mechanistic studies suggested that the cardiotonic effect of compound 5: is mediated by enhanced cellular calcium influx. Metabolites of TA with these structural features may be useful therapeutics to prevent heart failure.

Association between dietary fatty acid patterns based on principal component analysis and fatty acid compositions of serum and breast milk in lactating mothers in Nanjing, China.

This study aimed to comprehensively analyze dietary fatty acids (FAs) to evaluate their association with FA compositions of maternal serum and breast milk and assess their effects on mothers and infants. Overall, 121 healthy lactating Chinese mothers at 30-50 days of postpartum were enrolled and instructed to complete a Food Frequency Questionnaire, together with venous blood and breast milk sample collections. Dietary FA patterns were derived by principal component analysis with varimax rotation. Serum and breast milk FA compositions were detected using capillary gas chromatography and presented as relative concentrations (weight percentage of total FAs, %). Daily energy intake, absolute intake of most nutrients, and percentage of energy intake provided by these nutrients significantly varied among the different dietary FA patterns. There were significant differences in serum polyunsaturated fatty acid (PUFA) levels (P = 0.011); in monounsaturated fatty acid and PUFA proportions in breast milk with respect to four patterns (P = 0.002 and P = 0.026, respectively); and in n-6 PUFA, n-3 PUFA, linoleic acid, γ-linolenic acid, α-linolenic acid, and docosahexaenoic acid levels in breast milk (P = 0.027, P = 0.007, P = 0.048, P = 0.034, P = 0.020, and P = 0.002, respectively). Furthermore, maternal weight retention and length-for-age z scores, weight-for-age z scores and head circumference-for-age z scores of infants with respect to the four patterns exhibited significant differences (P = 0.038, P = 0.030, P = 0.034, and P<0.001, respectively). The results demonstrated the effect of dietary FA patterns on FA compositions of serum and breast milk, and patterns mainly characterized by LC-PUFA may have potentially beneficial effects on maternal postpartum recovery and infant growth.

Qigong for women with breast cancer: An updated systematic review and meta-analysis.

OBJECTIVE: The purpose of this review was to evaluate the effectiveness of Qigong in improving the quality of life and relieving fatigue, sleep disturbance, and cancer-related emotional disturbances (distress, depression, and anxiety) in women with breast cancer. METHODS: The PubMed, Cochrane Central Register of Controlled Trials, Web of Science, Sinomed, Wanfang, VIP, and China National Knowledge Infrastructure databases were searched from their inceptions to March 2020 for controlled clinical trials. Two reviewers selected relevant trials that assessed the benefit of Qigong for breast cancer patients independently. A methodological quality assessment was conducted according to the criteria of the 12 Cochrane Back Review Group for risk of bias independently. A meta-analysis was performed by Review Manager 5.3. RESULTS: This review consisted of 17 trials, in which 1236 cases were enrolled. The quality of the included trials was generally low, as only five of them were rated high quality. The results showed significant effectiveness of Qigong on quality of life (n = 950, standardized mean difference (SMD), 0.65, 95 % confidence interval (CI) 0.23-1.08, P =  0.002). Depression (n = 540, SMD = -0.32, 95 % CI -0.59 to -0.04, P =  0.02) and anxiety (n = 439, SMD = -0.71, 95 % CI -1.32 to -0.10, P =  0.02) were also significantly relieved in the Qigong group. There was no significant benefit on fatigue (n = 401, SMD = -0.32, 95 % CI  0.71 to 0.07, P = 0.11) or sleep disturbance relief compared to that observed in the control group (n = 298, SMD = -0.11, 95 % CI  0.74 to 0.52, P = 0.73). CONCLUSION: This review shows that Qigong is beneficial for improving quality of lifeand relieving depression and anxiety; thus, Qigong should be encouraged in women with breast cancer.

Long-term supplementation of dehydroepiandrosterone improved depressive-like behaviors by increasing BDNF expression in the hippocampus in ovariectomized rats.

OBJECTIVE: Dehydroepiandrosterone (DHEA), a precursor of estrogen, partially exhibits its biological effect after conversion to estrogen. Its biological significance in perimenopausal depressive disorder or postpartum depression remains unknown. Here, we observed the effects of long-term supplementation of DHEA on depression-like behaviors in ovariectomized rats. METHODS: We established the model as one of sex hormone deficiency in female rats by bilateral ovariectomy. We observed the effects of 13.3 mg/kg DHEA or 0.27 mg/kg estradiol were given daily by gavage for 12 weeks on lipid metabolism, glucose tolerance, and depression-like behaviors in ovariectomized rats. Furthermore, the expression of brain-derived neurotrophic factor (BDNF) and its signaling molecule in the hippocampus was analyzed. RESULTS: The 12-week supplementation of DHEA or estradiol significantly alleviated weight gain and improved the glucose tolerance in the ovariectomized rats. Moreover, Long-term supplement of DHEA or estradiol significantly increased sucrose preference and locomotion activities, and reduced immobility duration of the ovariectomized rats in the water. Both DHEA and estradiol treatments increased the expression of BDNF, phosphorylation of ERK and CREB, and ERß, but not that of ERα in the hippocampus of the ovariectomized rats. CONCLUSIONS: Overall, chronic treatment with DHEA improved depression-like behaviors in ovariectomized rats, suggesting that it may be useful for the treatment of sex hormone deficiency such as perimenopausal depressive disorder or postpartum depression.

Sijunzi decoction may decrease apoptosis via stabilization of the extracellular matrix following cerebral ischaemia-reperfusion in rats.

Neurons undergo degeneration, apoptosis and death due to ischaemic stroke. The present study investigated the effect of Sijunzi decoction (SJZD), a type of traditional Chinese medicine known as invigorating spleen therapy, on anoikis (a type of apoptosis) in rat brains following cerebral ischaemia-reperfusion. Rats were randomly divided into sham, model, nimodipine and SJZD low/medium/high dose groups. A middle cerebral artery occlusion model was established. Neurobehavioural scores were evaluated after administration for 14 days using a five-grade scale. Blood-brain barrier permeability and apoptotic rate were detected using Evans blue (EB) extravasation and TUNEL staining, respectively. Tissue inhibitor of metalloproteinase 1 (TIMP-1), matrix metalloproteinase 9 (MMP-9) and collagen IV (COL IV) were determined using immunohistochemistry. Neurobehavioural scores decreased remarkably in all SJZD and nimodipine groups compared to the model group (P<0.05). Compared with the sham group, EB extravasation was higher in the model group (P<0.01). The amount of EB extravasation decreased in the SJZD high dose and nimodipine groups compared to the model group (P<0.01), and extravasation in the SJZD high dose group was lower than the SJZD low and medium dose groups (P<0.01). TIMP-1 and MMP-9 expression and apoptotic rate increased, but COL IV decreased significantly in the hippocampus of the model group compared to the sham group (P<0.01). TIMP-1 and COL IV expression increased significantly and MMP-9 and apoptotic rate decreased remarkably in all SJZD and nimodipine groups compared to the model group (P<0.01). TIMP-1 and COL IV expression decreased, but MMP-9 expression and apoptotic rate increased in the SJZD low and medium dose groups compared to the SJZD high dose group (P<0.01). SJZD rescued neurons and improved neurobehavioural function in rats following cerebral ischaemia-reperfusion, especially when used at a high dose. The mechanism may be related to protection of the extracellular matrix followed by anti-apoptotic effects.

Absorptive characteristic of Bletilla striata extract in Caco-2 cell monolayer / 中国中药杂志

To investigate the active fraction from Bletilla striata in Caco-2 cell monolayer,so as to explore its absorption mechanism of oral administration preliminarily.Active fraction from B.striata in Caco-2 cell monolayer was analyzed by UPLC-Q-TOF and detected by UPLC-MS/MS,and the effects of different concentrations,pH and P-glycoprotein inhibitors on Caco-2 cells Monolayer were investigated.Six compounds were isolated from the active fraction of B.striata in Caco-2 cell monolayer by UPLC-Q-TOF,and identified as B6,B12,B14,B17,B19 and B23,with concentration dependence.Within the 0-180 min,the uptake of B12 and B14 had a time dependence,while B6,B17,B19 and B23 tended to saturate after 60 min.All of the components had a good absorption in an acidic environment.B6 had a good absorption at pH 6.0,while the other components B12,B14,B17,B19 and B23 had a good absorption at pH4.0.The absorption of the 6 main components of B.striata were not be affected by P-glycoprotein inhibitors(verapamil/cyclosporin A).Compared with the control group,there was no difference in the absorption of B6 and B12,and the absorption of B14,B17,B19 and B23 increased,but with no significant difference.The absorption characteristic of B.striata extract across the Caco-2 cell monolayer is probably passive diffusion,and the absorption process was not affected by P-glycoprotein.

[Effect of Shenqi Dihuang decoction on inflammatory factor, renal function and microcirculation in patients with early diabetic nephropathy].

To study the effect of Shenqi Dihuang decoction on inflammatory factor, renal function and microcirculation in patients with early diabetic nephropathy. A total of 205 cases of patient with early diabetic nephropathy treated in our hospital from March 2014 to April 2016 were selected and divided into two groups according to the admitted order, with 103 cases in clinical group and 102 in control group. Patients in control group were treated with melbine and captopril, which may be adjusted according to the clinical symptom. The clinical group was given Shenqi Dihuang decoction. Then the clinical efficady, inflammatory factors, renal function, endothelial function and hemorheology index were compared. Compared with 77.45% in the control group, the total effective rate of the clinical group was 92.23%. There was a significant increase (P<0.05). The comparison of the values of IL-6, IL-8, TNF-α and CRP between the two groups before and after treatment showed significant differences. The values of inflammatory factors in treatment group were lower than in control group (P<0.05). The comparison of the values of ß2-MG, Cys-C and urine m-ALB between the two groups before and after treatment showed significant differences. The values of renal function in treatment group were lower than those in control group (P<0.05). Compared with before treatment, ET-1 of the two groups after treatment decreased, while NO increased (P<0.05). Compared with the control group, the value of ET-1 in patients of the experimental group was lower after treatment, while NO was higher (P<0.05). The comparison of the values of whole blood viscosity, plasma viscosity, whole blood reduction viscosity, platelet aggregation rate and fibrinogen between the two groups before and after treatment showed significant differences. The value of hemorheology index in treatment group was lower than that in control group (P<0.05). Shenqi Dihuang decoction has a better effect on patients with early diabetic nephropathy. It can significantly intervene with inflammatory response, reduce proteinuria, protect the renal function of patients, and improve the patient's vascular endothelial function and blood rheology, so as to make microcirculation to recover to the normal level.

Environmental Benefit Assessment for the Carbonation Process of Petroleum Coke Fly Ash in a Rotating Packed Bed.

A high-gravity carbonation process was deployed at a petrochemical plant using petroleum coke fly ash and blowdown wastewater to simultaneously mineralized CO2 and remove nitrogen oxides and particulate matters from the flue gas. With a high-gravity carbonation process, the CO2 removal efficiency was found to be 95.6%, corresponding to a capture capacity of 600 kg CO2 per day, at a gas flow rate of 1.47 m3/min under ambient temperature and pressure. Moreover, the removal efficiency of nitrogen oxides and particulate matters was 99.1% and 83.2%, respectively. After carbonation, the reacted fly ash was further utilized as supplementary cementitious materials in the blended cement mortar. The results indicated that cement with carbonated fly ash exhibited superior compressive strength (38.1 ± 2.5 MPa at 28 days in 5% substitution ratio) compared to the cement with fresh fly ash. Furthermore, the environmental benefits for the high-gravity carbonation process using fly ash were critically assessed. The energy consumption of the entire high-gravity carbonation ranged from 80 to 169 kWh/t-CO2 (0.29-0.61 GJ/t-CO2). Compared with the scenarios of business-as-usual and conventional carbon capture and storage plant, the economic benefit from the high-gravity carbonation process was approximately 90 and 74 USD per ton of CO2 fixation, respectively.

[Changes of perimysial junctional plates induced by excessive eccentric training and the effects of acupuncture intervention].

This study aimed to investigate the effects of acupuncture intervention on excessive eccentric training-induced changes of perimysial junctional plates (PJPs) domain. Thirty healthy male Wistar rats were randomly assigned to 5 groups: control group, four-week training group, four-week training + 1-week recovery group and four-week training + 1-week acupuncture group. Rats were subjected to continuous excessive eccentric training for 4 weeks (incline -16°, speed 16-20 m/min, 60-90 min/d, 5 day per week), and then were subjected to one-week spontaneous recovery or one-week recovery with acupuncture intervention (a piece of filiform needle for 4 min every day). The PJPs domain changes were observed under transmission electron microscopy, and the perimysial collagen network structural changes were examined by scanning electron microscopy with or without a digestion technique (NaOH). The following results were obtained: (1) Compared with control group, PJPs domain of four-week training group showed excessive shortening of sarcomere (P < 0.001), serious damage of sarcomere structure, and altered mitochondria morphology in intermyofibria and subsarcolemma; 54% degradation of sarcolemma, and increased number of caveolae (P < 0.01); reduced number of PJPs (P < 0.001). (2) In comparison with four-week training group, PJPs domain was slightly changed in four-week training + 1-week recovery group, i.e., partial recovery of sarcomere length and structure (accounting for 85.23% of control group), and recovery of intermyofibrial and subsarcolemmal mitochondria morphology; decreased sarcolemmal degradation (P < 0.001), and increased number of caveolae (P < 0.05); increased PJPs number (P < 0.001). (3) PJPs domain changed in four-week training + 1-week acupuncture group compared with four-week training + 1-week recovery group, which were substantial recovery of sarcomere length (accounting for 94.51% of control group), increased subsarcolemmal mitochondrial fusion (P < 0.001), decreased caveolae number (P < 0.001), and decreased PJPs number (P < 0.001). The results indicated that excessive eccentric training resulted in excessively reduced number of PJPs with altered PJPs domain homeostasis, thus impeding the adaptability to eccentric training. After 1 week of natural recovery, the number of PJPs was excessively increased, hindering muscle damage repair. Acupuncture intervention helped to recover PJPs number and PJPs domain homeostasis, thus significantly relieving overuse injuries.

[External therapy of plasma cell mastitis by jiuyi powder using partial least-squares discriminant analysis: a safety analysis].

OBJECTIVE: To evaluate the safety and the clinical value of external use of jiuyi Powder (JP) in treating plasma cell mastitis using partial least-squares discriminant analysis (PLSDA). METHODS: Totally 50 patients with plasma cell mastitis treated by external use of JP were observed and biochemical examinations of blood and urine detected before application, at day 4 after application, at day 1 and 14 after discontinuation. Blood mercury and urinary mercury were detected before application, at day 1, 4, and 7 after application, at day 1 and 14 after discontinuation. Urinary mercury was also detected at 28 after discontinuation and 3 months after discontinuation. The information of wound, days of external application and the total dosage of external application were recorded before application, at day 1, 4, and 7 after application, as well as at day 1 after discontinuation. Then a discriminant model covering potential safety factors was set up by PLSDA after screening safety indices with important effects. The applicability of the model was assessed using area under ROC curve. Potential safety factors were assessed using variable importance in the projection (VIP). RESULTS: Urinary ß2-microglobulin (ß2-MG), urinary N-acetyl-ß-D-glucosaminidase (NAG), 24 h urinary protein, and urinary α1-microglobulin (α1-MG) were greatly affected by external use of JP in treating plasma cell mastitis. The accuracy rate of PLSDA discriminate model was 74. 00%. The sensitivity, specificity, and the area under ROC curve was 0. 7826, 0. 7037, and 0. 8084, respectively. Three factors with greater effect on the potential safety were screened as follows: pre-application volume of the sore cavity, days of external application, and the total dosage of external application. CONCLUSIONS: PLSDA method could be used in analyzing bioinformation of clinical Chinese medicine. Urinary ß2-MG and urinary NAG were two main safety monitoring indices. Days of external application and the total dosage of external application were main factors influencing blood mercury and urine mercury. A safety classification simulation model of treating plasma cell mastitis by external therapy of JP was established by the two factors, which could be used to assess the safety of external application of JP to some extent.

Yiqi formula enhances the antitumor effects of erlotinib for treatment of triple-negative breast cancer xenografts.

Yiqi formula (YF), a traditional herbal prescription, has long been used to treat triple-negative breast cancer (TNBC) patients. The present study aims to investigate the effects and the related mechanism of YF for treatment of TNBC xenografts. MDA-MB-231 (human TNBC) cells were subcutaneously injected into the second mammary fat pad of 40 female nude mice, which were divided into four groups: control, erlotinib (an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor), YF, and combination (YF plus erlotinib). All treatments were administered orally for 30 days. Inhibition rate of tumor weight by erlotinib, YF, and the combination was 26.47%, 17.24%, and 39.15%, respectively. Western blotting showed that YF, erlotinib, and the combination downregulated p-EGFR (P < 0.01) and p-Akt1 (pT308) (P < 0.05) and upregulated PTEN compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). Similar results were detected by immunohistochemistry. Real-time quantitative PCR showed that YF, erlotinib, and the combination increased PTEN mRNA (P < 0.05, P < 0.01) compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). In conclusion, YF can regulate the main components of the PI3K/Akt pathway in TNBC xenografts. When YF was used in combination with erlotinib, it enhanced the antitumor effects of erlotinib on TNBC xenografts. These findings suggest that YF is suitable to use for the treatment of TNBC patients.

An old herbal medicine with a potentially new therapeutic application in inflammatory bowel disease.

Inflammatory Bowel Disease (IBD) is a chronic and frequently disabling inflammatory disorder of the intestine. New developments in IBD therapy are primarily focused on biologic treatments; however, they are both expensive and associated with significant side effects. Here, we provide the first preclinical evidence that YunNan BaiYao (YNBY), a well-known traditional Chinese herbal remedy frequently used for treating hemorrhages and wounds, can effectively alleviate experimental colitis. Oral administration of YNBY in drinking water significantly reduced the disease activities of both DSS- and TNBS-induced experimental colitis. Mechanistic studies revealed that the effectiveness of YNBY was not due to an anti-bacterial function since YNBY had no effect on E. coli growth. Rather, it exhibited an anti-inflammatory or immunosuppressive function: In the DSS colitis model, YNBY treatment decreased the levels of several pro-inflammatory cytokines in colonic mucosa, including TNFα, IL-12p40, and IL-17. Similar cytokine changes were also observed in mouse serum, suggesting that systemic changes in general reflect the changes in the affected colon. Significant down-regulation of IL-12p40 and IL-17, in addition to IFNγ, was also seen in TNBS-colitis model. Another potential mechanism for the anti-inflammatory effects of YNBY involves the selective suppression of pro-inflammatory immune cells: YNBY effectively suppressed the growth of multiple T- and B-lymphocytes, including Molt-4, Jurkat, and EBV-transformed human B-lymphocytes, more potently than 6-mecaptopurine (6-MP) and 5-aminosalicylic acid (5-ASA), two of the most commonly used first-line drugs in IBD therapy. In sharp contrast, YNBY exhibited no cytotoxicity to colonic epithelial cells (Caco-2 cells), even at the concentration 10-fold higher than that used in the lymphocyte model; and instead promoted cell spreading and wound healing. These results strongly suggest that YNBY not only has effective anti-inflammatory properties through suppressing lymphocyte growth and pro-inflammatory cytokine expression, but also can promote intestinal epithelial wound-healing and repair. Therefore, YNBY demonstrates strong potential as an alternative herbal therapy for IBD.

[Effects of Astragalus polysaccharide on proliferation and Akt phosphorylation of the basal-like breast cancer cell line].

OBJECTIVE: To investigate the effects of Astragalus polysaccharide (APS) on proliferation of basal-like breast cancer cell line MDA-MB-468 cells and Akt phosphorylation in MDA-MB-468 cells. METHODS: APS at different concentrations was used to culture MDA-MB-468 cells for different time periods, and then proliferation of MDA-MB-468 cells was assayed using methyl thiazolyl tetrazolium (MTT) assay to determine the time- and dose-dependent effects of APS. For observing the effects of APS on phosphor-Akt (p-Akt), in-cell Western blot method was used after 1, 2, 4 and 7 d of culture in APS to detect protein expressions of p-Akt (Thr308) and p-Akt (Ser473). Protein levels of the key targets in p53/murine double minute 2 (MDM2) signaling pathway, such as p53, MDM2 and phosphatase and tensin homolog deleted on chromosome ten (PTEN) were also detected. After PTEN gene was silenced by small interfering RNA (siRNA) in MDA-MB-468 cells, expressions of p-Akt (Thr308 and Ser473) were assayed by the in-cell Western blot method after 2 d of APS treatment. RESULTS: APS at 1 and 0.5 mg/mL concentrations effectively inhibited the proliferation of MDA-MB-468 cells and was used in subsequent tests. Compared with the control group, APS decreased the protein expression of p-Akt (Thr308) in MDA-MB-468 cells after 1-, 2-, 4- and 7-day culture, and also decreased the protein expression of p-Akt (Ser473) and up-regulated the protein expression of MDM2 in MDA-MB-468 cells after 1- and 2-day culture. Expressions of p53 and PTEN were up-regulated after 7 d of APS culture. After silencing PTEN gene by siRNA, APS could not mediate Akt phosphorylation. CONCLUSION: APS can inhibit proliferation of basal-like breast cancer cell line MDA-MB-468, and down-regulate the expression of Akt phosphorylation. The antiproliferation mechanisms may be related to its effects of up-regulating the expressions of p53 and PTEN by regulating p53/MDM2 positive and negative feedback loops.

[Study protocol for a self-controlled case study to evaluate the safety and standardization for external application of Chinese medicine Jiuyi Powder].

BACKGROUND: As the main medicinal powder for drawing out pus and removing necrotic tissue in external therapies of traditional Chinese surgery, Sheng Powder has made great contributions to the treatment of inflammatory wounds and has the unique bactericidal and decay-discharging function that can not be replaced by antibiotics. However, Sheng Powder has toxicity because it contains mercury. So far, there is no clinical research on the standards of dose and usage of Sheng Powder and there is a lack of objective and quantitative criteria for operating standards and monitoring of toxicity and side effects. Therefore, the authors choose Jiuyi Powder, one of the most commonly used Sheng Powder, to evaluate the safety of its external use, and form a standardization program for clinical implementation. METHODS AND DESIGN: This study is a nonrandomized, nonblinded, self-controlled case study. There will be two key stages in the study. In stage one, 10 patients with plasma cell mastitis will be enrolled. The patients will receive continuous external application of Jiuyi Powder with a fixed dose (160 mg/d). Blood mercury and urine mercury levels will be dynamically tested at different time intervals, and the observation of pharmacokinetic parameters will be conducted after Jiuyi Powder has been absorbed by the surface of the wound. In stage two, 30 patients with each of the four conditions including postoperative wounds of head or facial surgeries, plasma cell mastitis, anal fistula and chronic absorption ulcer of the shank will be enrolled, respectively. According to the dose of 1.5 mg/cm(2), Jiuyi Powder will be externally applied to the wound surface for 14 d. On the basis of the first-phase study, the test time will be chosen and the toxicity outcome will be detected to evaluate the safety of external application of Jiuyi Powder and to establish recommendations for standardized clinical use. DISCUSSION: The purpose of this study is to evaluate the safety of the external application of Jiuyi Powder through a two-stage study. The pharmacokinetic parameters of external application of Jiuyi Powder at the clinical dose range, the changes in blood and urinary mercury levels and related safety indexes will be observed on different wounds located in different positions externally. The evaluation of whether Jiuyi Powder could be a routine medication for drawing out pus and removing necrotic tissue in inflammatory and refractory wounds will be stated. TRIAL REGISTRATION NUMBER: ChiCTR-TNC-11001366.

Quantification of total iodine in intact granular starches of different botanical origin exposed to iodine vapor at various water activities.

Iodine has been used as an effective tool for studying both the structure and composition of dispersed starch and starch granules. In addition to being employed to assess relative amylose contents for starch samples, it has been used to look at the molecular mobility of the glucose polymers within intact starch granules based on exposure to iodine vapor equilibrated at different water activities. Starches of different botanical origin including corn, high amylose corn, waxy corn, potato, waxy potato, tapioca, wheat, rice, waxy rice, chick pea and mung bean were equilibrated to 0.33, 0.75, 0.97 water activities, exposed to iodine vapor and then absorbance spectra and LAB color were determined. In addition, a new iodine quantification method sensitive to <0.1% iodine (w/w) was employed to measure bound iodine within intact granular starch. Amylose content, particle size distribution of granules, and the density of the starch were also determined to explore whether high levels of long linear glucose chains and the surface area-to-volume ratio were important factors relating to the granular iodine binding. Results showed, in all cases, starches complexed more iodine as water content increased and waxy starches bound less iodine than their normal starch counterparts. However, much more bound iodine could be measured chemically with waxy starches than was expected based on colorimetric determination. Surface area appeared to be a factor as smaller rice and waxy rice starch granules complexed more iodine, while the larger potato and waxy potato granules complexed less than would be expected based on measured amylose contents. Corn, high amylose corn, and wheat, known to have starch granules with extensive surface pores, bound higher levels of iodine suggesting pores and channels may be an important factor giving iodine vapor greater access to bind within the granules. Exposing iodine vapor to moisture-equilibrated native starches is an effective tool to explore starch granule architecture.

[Effects of Astragalus injection on proliferation of basal-like breast cancer cell line MDA-MB-468].

OBJECTIVE: To investigate the effects of Astragalus injection (AI) on basal-like breast cancer cell line MDA-MB-468 and murine bone marrow stromal stem cells (mMSCs). METHODS: MDA-MB-468 cells and primary cultured mMSCs were treated by different concentrations of AI, and with untreated MDA-MB-468 cells as blank control. The morphology of cells was observed by phase-contrast inverted microscope and transmission electron microscopy. Cytotoxic effects of AI on MDA-MB-468 cells and mMSCs were evaluated by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis of MDA-MB-468 cells induced by AI were measured by flow cytometry. Lactate dehydrogenase (LDH) activity in supernatants was measured by enzymatic colorimetric method. The expressions of epidermal growth factor receptor (EGFR) and p53 protein in MDA-MB-468 cells were evaluated by streptavidin-biotin-peroxidase complex method. RESULTS: A time-dependent cytotoxic effect of 1 g/ml AI was observed in MDA-MB-468 cells. 1 g/ml AI also had cytotoxic effect on mMSCs, but its effect was not better than cisplatin. 0.1 g/ml AI could promote the proliferation of mMSCs. Different concentrations of AI could all induce the apoptosis of MDA-MB-468 cells. There was no significant difference in LDH activity in the supernatants between blank control group and AI-treated and cisplatin-treated groups. AI could down-regulate the expressions of EGFR and p53 protein. CONCLUSION: The effects of AI on MDA-MB-468 cells and mMSCs are related to the concentration of AI, and its mechanism of inhibiting the proliferation of MDA-MB-468 cells may be due to down-regulation of the expressions of EGFR and p53 protein.

[Effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture].

OBJECTIVE: To evaluate the effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture. METHODS: The normal human mammary epithelial cells were dissociated by digestion with collagenase type I. The morphological identification of normal human mammary epithelial cells in primary culture was determined under inverted phase contrast microscope and transmission electron microscope. The estradiol and progesterone were added to culture medium to induce the proliferation of normal human mammary epithelial cells in primary culture, and the different doses of Chinese herbal medicine for warming kidney and tamoxifen were also added into the culture medium. The MTT assay was used to evaluate the proliferation of normal human mammary epithelial cells in primary culture under the different conditions of interventions. RESULTS: The normal human mammary epithelial cells in primary culture presented typically morphological features of normal human mammary epithelial cells in vitro. The proliferation of normal human mammary epithelial cells in primary culture was significantly improved by intervention of estradiol and progesterone both with concentration of 1x10(-5) g/L. The high-dose Chinese herbal medicine for warming kidney could obviously promote the proliferation of normal human mammary epithelial cells in primary culture, while the proliferation was obviously inhibited by low-dose Chinese herbal medicine for warming kidney. The tamoxifen exerted no effects on the proliferation of normal human mammary epithelial cells in primary culture. CONCLUSION: The estradiol and progesterone can enhance the ability of proliferation of normal human mammary epithelial cells in primary culture. The regulating effect of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture is dose-dependent.