Choline induced cardiac dysfunction by inhibiting the production of endogenous hydrogen sulfide in spontaneously hypertensive rats.

To investigate the exact effects of dietary choline on hypertensive heart disease (HHD) and explore the potential mechanisms, male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were randomly divided into five groups as follows: WKY group, WKY + Choline group, SHR group, SHR + Choline group, and SHR + Choline + NaHS group. In choline treatment groups, rats were fed with 1.3% (w/v) choline in the drinking water for 3 months. The rats in the SHR + Choline + NaHS group were intraperitoneally injected with NaHS (100 micromol/kg/day, a hydrogen sulfide (H2S) donor) for 3 months. After 3 months, left ventricular ejection fraction (LVEF) and fractional shortening (LVFS), the indicators of cardiac function measured by echocardiography, were increased significantly in SHR as compared to WKY, although there was no significant difference in collagen volumes and Bax/Bcl-2 ratio between the two groups, indicating the early stage of cardiac hypertrophy. There was a significant decrease in LVEF and LVFS and an increase in collagen volumes and Bax/Bcl-2 ratio in SHR fed with choline, meanwhile, plasma H2S levels were significantly decreased significantly in SHR fed with choline accompanying by the decrease of cystathionine-gamma-lyase (CSE) activity. Three months of NaHS significantly increased plasma H2S levels, ameliorated cardiac dysfunction and inhibited cardiac fibrosis and apoptosis in SHR fed with choline. In conclusion, choline aggravated cardiac dysfunction in HHD through inhibiting the production of endogenous H2S, which was reversed by supplementation of exogenous H2S donor.

[Reflections on Chinese residents' calcium intake and human health].

Calcium plays an important role in various physiological activities of the human body, and long-term insufficient or excessive intake of calcium will have a negative impact on the body's health. Existing data show that insufficient dietary calcium intake is closely related to bone health, but the non-bone effects are not clear. Increasing dietary calcium and supplementing calcium (with or without vitamin D) have a certain beneficial effect on the bone mineral density and its peak of adolescents and can delay the bone loss of the elderly, but it can't improve the height and bone mineral density of adults and fracture in the elderly. This article introduces the physiological functions of calcium, dietary sources, human intake, and methods for formulating recommended amounts, and summarizes the relationship between calcium and health effects. It also recommends that when formulating the reference intake of dietary calcium for Chinese residents, more consideration should be given to the data and information of the Chinese population, combined with the characteristics of Chinese residents' genetics, absorption and metabolism.

[Effects of different calcium intakes on metabolism in healthy adults under strict dietary intervention].

Objective: To explore the effect of different calcium intakes on metabolism in healthy adults with traditional Chinese dietary pattern. Methods: A total of 64 college students (32 males and 32 females) were recruited from the School of Public Health, Harbin Medical University from September to October 2015. A prospective, randomized, double-blind, placebo-controlled trial design (RCT trial) was used to strictly control the diet of the volunteers through meal preparation, so that the background of the experimental and the control group was exactly the same. In our study, 64 healthy adults were randomly divided into two groups (calcium supplementation and control). The two groups were given the same diet every day. One group was given 400 mg/d of calcium supplementation, which was close to the recommended intake in China (800 mg/d); the other group was given pacebo, the average calcium intake was close to the average intake of Chinese (400 mg/d), calcium intervention lasted for 35 days. Serum and urine samples from pre-intervention, mid-intervention, and post-intervention were collected for metabolomics studies and detection of serum hormone levels and biochemical indicators. PCA and PLS-DA were used to compare the differences in serum and urine metabolites. After adjusting for age and BMI, the differences of hormones [parathyroid hormone, 1, 25(OH)2D3] and biochemical indicators were compared by repeated measurement variance analysis. Results: The 64 subjects were aged 23-28 years, including 32 males and 32 females. The baseline characteristics of the calcium supplementation group and the control group were balanced. The results showed that there were no significant differences in blood and urine metabolites, metabolic hormone [parathyroid hormone, 1, 25(OH)2D3] levels and biochemical indicators(serum Ca, serum P, glycolipid metabolism and hepatorenal function indicators)between the calcium supplementation group and control group at each time point throughout the trail (P>0.05). Conclusion: No health hazards associated with calcium deficiency was observed in healthy adults in the short term at current calcium intake level (300-400 mg/d).

Effects of taurine on growth performance, antioxidant capacity, and lipid metabolism in broiler chickens.

To investigate the effects of dietary taurine supplementation on growth performance, antioxidant status, and lipid metabolism in broilers, 384 male broilers (Arbor Acres, 1 D of age) were randomly allocated into 4 groups with 8 replicates of 8 birds. Dietary treatments were supplemented with taurine at the level of 0.00, 2.50, 5.00, and 7.50 g/kg of the diet (denoted as CON, TAU1, TAU2, TAU3, respectively). The BW gain from 1 to 21 D and from 22 to 42 D were all increased linearly (linear, P < 0.001) by taurine supplementation. Throughout the trial period, the highest BW gain and favorable gain-to-feed ratio were observed in the TAU2 group. Taurine supplementation increased the antioxidant enzyme activities and decreased (linear, P < 0.001) the content of malondialdehyde in both serum and the liver of broilers and alleviated oxidative damage through enhancing (P < 0.05) the hepatic genes expression of nuclear factor erythroid-2-related factor 2 (NRF2), glutathione peroxidase (GPX), and heme oxygenase-1 (HO-1). Correspondingly, in serum, the activities of hepatic lipase and total lipase were decreased linearly and quadratically (linear and quadratic, P < 0.001) with the increasing inclusion of taurine in the diet. Meanwhile, in serum, the content of triglycerides was significantly decreased (P < 0.05), and except for TAU3, the total cholesterol content was also significantly decreased (P < 0.05) by taurine supplementation. In addition, the hepatic content of triglycerides was significantly decreased (P < 0.05) in the TAU1 and TAU2 groups. Compared with the CON group, the hepatic genes expression of adenosine monophosphate-activated protein kinase alpha (AMPKα), silent 1, (SIRT1) and carnitine palmitoyltransferase 1 (CPT-1) were all increased (P < 0.05), and sterol regulatory element-binding protein-1 (SREBP-1) expression was decreased (P < 0.05) in the TAU2 group. These results indicated that taurine supplementation improved the growth performance, antioxidant capacity, and lipid metabolism of broilers.

Transfemoral Transcatheter Aortic Valve Replacement Using Fascia Iliaca Block as an Alternative Approach to Conscious Sedation as Compared to General Anesthesia.

BACKGROUND/PURPOSE: General Anesthesia (GA) and conscious sedation (CS) are anesthetics for transfemoral transcatheter aortic valve replacement (TF-TAVR). We compared TF-TAVR outcomes using a novel anesthetic approach with fascia iliaca block (FIB) plus minimal CS (MCS) versus GA. METHODS: This retrospective propensity-matched study included consecutive TF-TAVR patients from January 2013 to December 2017 and dichotomized into FIB-MCS vs. GA. Data were collected from electronic records, Society of Thoracic Surgery (STS) database, and the Transcatheter Valve Therapies (TVT) Registry. Primary endpoints were operating room (OR) time, intensive care unit (ICU) and hospital length of stay (LOS). Secondary endpoints were 30-day, 1-year mortality, quality of life, 30-day re-hospitalization rate, failure of FIB-MCS, and hospital safety outcomes. RESULTS: A total of 304 TF-TAVR patients; FIB-MCS (n = 219) vs. GA (n = 85). Propensity matched 162 patients; FIB-MCS (n = 108) vs. GA (n = 54). FIB-MCS had shorter OR time (197.6 ±â€¯56.3 vs. 248.2 ±â€¯46.3 min, p < 0.001), ICU (67.8 ±â€¯71.7 vs. 84.9 ±â€¯72.1 h, p = 0.004) and hospital LOS (3.2 ±â€¯3.7 vs. 5.9 ±â€¯3.5 d, p < 0.001). FIB-MCS had lower rate of blood transfusion. FIB-MCA vs. GA 30-day and 1-year mortality were similar in the entire (2.3 vs. 2.4%, p = 1.0; and 8.2 vs. 5.9%, p = 0.49) and matched cohorts (0 vs. 3.7%, p = 0.11 and 7.4 vs. 5.6%, p = 0.75). FIB-MCS were less likely to be re-hospitalized [Odd Ratio: 0.32, CI:0.13-0.76] and 2% to 3% higher KCCQ-12 score. CONCLUSION: TF-TAVR using FIB-MCS is feasible and safe with shorter OR time, ICU and hospital LOS, lower risk of 30-day re-hospitalization, similar 30-day and 1-year mortality with better quality of life at 1-year follow-up.

The prevalence, characteristics, expenditure and predictors of complementary medicine use in Australians living with gastrointestinal disorders: A cross-sectional study.

AIMS: To determine the prevalence, characteristics, expenditure and predictors of complementary medicine (CM) use in Australian adults living with gastrointestinal disorders (GID). METHODS: A cross-sectional study involving 2,025 Australian adults was conducted. Participants were recruited through purposive convenience sampling. Descriptive statistics were conducted to report the prevalence of people living with GIDs and their CM use, including CM products, mind-body practices and CM practitioner services. Chi-square test and independent-samples t-test were used to determine the associations between sociodemographic or health-related variables with CM use. Binary logistic regression was conducted to determine the significant predictors of CM use in GID participants. Economic data was calculated based on the mean out-of-pocket expenditure on CM. RESULTS: Of the 293 participants reporting a GID, 186 (63.5%) used CM products, 55 (18.8%) used a mind-body practice and 141 (48.1%) visited at least one CM practitioner in the last 12 months. Collectively, the majority of GID participants using any type of CM were female, aged 40-49 years, married and employed. The mean score for health-related quality of life was 49.6 out of 100 in GID participants and 68.2 in participants without a GID (p < 0.001). Average annual out-of-pocket expenditure on CM products was AUD127.29 by CM products users with a GID. The predictors of CM products, mind-body practices and CM practitioner services use differed. Of the 111 CM product users with a GID, 103 (92.8%) disclosed all or some of their CM use to general practitioner, 89 (80.2%) to specialist doctor, 79 (71.2%) to pharmacist and 69 (62.1%) to hospital doctor. CONCLUSIONS: A substantial proportion of Australian adults living with GID use CM products, mind-body practices and CM practitioner services. This study provides important insights to inform and guide the development of a more coordinated health care services for individuals living with GID.

The prevalence and characteristics of complementary medicine use by Australian and American adults living with gastrointestinal disorders: A systematic review.

AIM: To report the prevalence and sociodemographic characteristics of complementary medicine (CM) use by Australian and American adults living with gastrointestinal (GI) disorders. METHOD: Cumulative Index to Nursing and Allied Health Literature (CINAHL), Embase, Medline, Pubmed, Scopus and Web of Science were searched for studies published in English language between 1st January 2008 and 8th April 2018. Additionally, a Google Scholar search and risk of bias assessment were conducted. RESULTS: Twelve (five American and seven Australian) studies, involving 53,991 adults were included. Data were extracted from studies reporting CM use by both specific populations with medically diagnosed GI disorders and the general population with self-reported GI disorders. CM use ranged from 23.6% to 44% (American) and 27.2% to 58% (Australian) by adults with medically diagnosed GI disorders. Whilst CM use was higher amongst females, with higher incomes and education levels. The prevalence of CM use by those with self-reported GI disorders varied from 0.31% to 80%. Herbal, probiotic supplements and mind-body practices were the most prevalent CMs used. The risk of bias assessment identified weaknesses in the external validity, principally related to poor study design. CONCLUSIONS: There is a substantial use of CMs by American and Australian adults with GI disorders. Females with higher education and income are predominant users. There is a need to conduct up-to-date and well-designed prevalence and sociodemographic studies. Additionally, future studies should consider including health literacy and medicines disclosure scales in order to better understand CM use by this population.

Effects of dietary supplementing tannic acid in the ration of beef cattle on rumen fermentation, methane emission, microbial flora and nutrient digestibility.

Four adult Simmental male cattle (376 ± 9.0 kg initial BW), fitted with permanent rumen cannulas, were used in a 4 × 4 Latin square design to investigate the effects of dietary supplementing tannic acid (TA) on rumen fermentation, methane (CH4 ) production, rumen microbes, nutrient digestibility and plasma biochemical parameters. Four levels of TA, that is 0, 6.5, 13.0 or 26.0 g/kg dry matter (DM), were added to the basal ration (composed of corn silage and concentrate mixture) as experimental treatments respectively. Each experimental period consisted of a 12-day adaptation phase followed by a 3-day sampling phase. The results showed that supplementing TA at 26.0 g/kg DM decreased the relative abundance of protozoa, methanogens and Ruminococcus albus to the total ruminal bacterial 16S rDNA in beef cattle (p < 0.05). The results also showed that supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) the CH4 production (l/kg DM intake) by 11.1%, 14.7% and 33.6% respectively. Supplementing TA at 13.0 or 26.0 g/kg DM decreased the ratio of acetate to propionate and ammonia nitrogen (NH3 -N) (p < 0.05) and tended to decrease the total volatile fatty acid (VFA) concentration of rumen fluid (p = 0.07). Supplementing TA at 26.0 g/kg DM decreased DM and organic matter (OM) digestibility (p < 0.05), supplementing TA at 6.5, 13.0 or 26.0 g/kg DM decreased (p < 0.01) crude protein (CP) digestibility by 5.0%, 8.6% and 15.7%, respectively, and supplementing TA at 6.5, 13.0 or 26.0 g/kg DM increased (p < 0.05) the plasma total antioxidant capability. It was concluded that supplementing TA in the ration of beef cattle decreased the CH4 production and digestibility of CP of beef cattle. Supplementing TA could be an effective option to mitigate CH4 emission form cattle, further research is necessary to study the effects of TA on the performance of cattle.

[Effectiveness of clinical pathway for treatment of advanced schistosomiasis hepatic fibrosis].

Objective To assess the effectiveness of the clinical pathway for the treatment of advanced schistosomiasis hepatic fibrosis. Methods The duration of hospital stay, gross hospitalization expense, individual-paid expense, interior diameter of portal vein, levels of four serum hepatic fibrosis-related parameters (PIIIP, CIV, HA, and LN), and activities of ALT, AST and γ-GT were assessed and compared between the advanced schistosomiasis patients receiving the clinical pathway and ones receiving non-clinical pathway. Results There were 142 advanced schistosomiasis patients with hepatic fibrosis receiving the clinical pathway of anti-hepatic fibrosis. Compared with the patients receiving non-clinical pathway, the gross hospitalization expenses reduced by 11.2% (t = 6.310, P < 0.05), and the individual-paid expenses reduced by 16.1% (t = 4.326, P < 0.05). The mean HA level was twice higher than the normal range, with a positive rising from 70.4% to 83.1%, and the abnormal rates of CIV and γ-GT were 64.1% and 28.9% respectively. Conclusions The clinical pathway can drastically reduce the treatment expenses in advanced schistosomiasis patients with hepatic fibrosis. However, the patients have a trend towards the persistent disease progression. Therefore, the researches of more effective therapeutic methods for advanced schistosomiasis hepatic fibrosis are urgently needed.

Development and significance of SCAR marker QG12-5 for Canarium album (Lour.) Raeusch by molecular cloning from improved RAPD amplification.

Sequence-characterized amplified region (SCAR) is a valuable molecular marker for the genetic identification of any species. This marker is mainly derived from molecular cloning of random amplified polymorphic DNA (RAPD). We have previously reported the use of an improved RAPD technique for the genetic characterization of different samples of Canarium album (Lour.) Raeusch (C. album). In this study, DNA fragments were amplified using improved RAPD amplified from different samples of C. album. The amplified DNA fragment was excised, purified from an agarose gel and cloned into a pGM-T vector; subsequently, a positive clone, called QG12-5 was identified by PCR amplification and enzymatic digestion and sequenced by Sanger di-deoxy sequencing method. This clone was revealed consisting of 510 nucleotides of C. album. The SCAR marker QG12-5 was developed using specifically designed PCR primers and optimized PCR conditions. This SCAR marker expressed seven continuous "TATG" [(TATG)n] tandem repeats, which was found to characterize C. album. Subsequently, this novel SCAR marker was deposited in GenBank with accession No. KT359568. Therefore, we successfully developed a C. album-specific SCAR marker for the identification and authentication of different C. album species in this study.

An improved DNA marker technique for genetic characterization using RAMP-PCR with high-GC primers.

Random amplified polymorphic DNA (RAPD) is a widely used molecular marker technique. As traditional RAPD has poor reproducibility and productivity, we previously developed an improved RAPD method (termed RAMP-PCR), which increased the reproducibility, number of bands, and efficiency of studies on polymorphism. To further develop the efficiency of this method, we used high-GC content primers for improved RAMP-PCR with DNA samples from Lonicera japonica. Comparison of amplification profiles obtained by standard RAPD primers with those obtained by regular PCR and RAMP-PCR, and high-GC primers with regular PCR and RAMP-PCR showed that the average number of bands and polymorphisms per primer gradually and significantly increased (from 6.4 to 15.0 and from 4.6 to 10.2, respectively). Cluster dendrograms showed similar results, indicating that this new method is consistent and reproducible. A total of 22 samples from different species, including plants, animals, and humans, were used for RAMP-PCR with high-GC primers. Multiple bands were successfully amplified from all samples, demonstrating that this method is a reliable technique with consistent results and may be of general interest in studies on different genera and species. We developed highly effective DNA markers, which can provide a more effective and potentially valuable approach than traditional RAPD for the genetic identification of various organisms, particularly of medicinal plants.

Development of novel SCAR markers for genetic characterization of Lonicera japonica from high GC-RAMP-PCR and DNA cloning.

Sequence-characterized amplified region (SCAR) markers were further developed from high-GC primer RAMP-PCR-amplified fragments from Lonicera japonica DNA by molecular cloning. The four DNA fragments from three high-GC primers (FY-27, FY-28, and FY-29) were successfully cloned into a pGM-T vector. The positive clones were sequenced; their names, sizes, and GenBank numbers were JYHGC1-1, 345 bp, KJ620024; YJHGC2-1, 388 bp, KJ620025; JYHGC7-2, 1036 bp, KJ620026; and JYHGC6-2, 715 bp, KJ620027, respectively. Four novel SCAR markers were developed by designing specific primers, optimizing conditions, and PCR validation. The developed SCAR markers were used for the genetic authentication of L. japonica from its substitutes. This technique provides another means of developing DNA markers for the characterization and authentication of various organisms including medicinal plants and their substitutes.

Targeting glutamine metabolism in multiple myeloma enhances BIM binding to BCL-2 eliciting synthetic lethality to venetoclax.

Multiple myeloma (MM) is a plasma cell malignancy that is largely incurable due to development of resistance to therapy-elicited cell death. Nutrients are intricately connected to maintenance of cellular viability in part by inhibition of apoptosis. We were interested to determine if examination of metabolic regulation of BCL-2 proteins may provide insight on alternative routes to engage apoptosis. MM cells are reliant on glucose and glutamine and withdrawal of either nutrient is associated with varying levels of apoptosis. We and others have demonstrated that glucose maintains levels of key resistance-promoting BCL-2 family member, myeloid cell leukemic factor 1 (MCL-1). Cells continuing to survive in the absence of glucose or glutamine were found to maintain expression of MCL-1 but importantly induce pro-apoptotic BIM expression. One potential mechanism for continued survival despite induction of BIM could be due to binding and sequestration of BIM to alternate pro-survival BCL-2 members. Our investigation revealed that cells surviving glutamine withdrawal in particular, enhance expression and binding of BIM to BCL-2, consequently sensitizing these cells to the BH3 mimetic venetoclax. Glutamine deprivation-driven sensitization to venetoclax can be reversed by metabolic supplementation with TCA cycle intermediate α-ketoglutarate. Inhibition of glucose metabolism with the GLUT4 inhibitor ritonavir elicits variable cytotoxicity in MM that is marginally enhanced with venetoclax treatment, however, targeting glutamine metabolism with 6-diazo-5-oxo-l-norleucine uniformly sensitized MM cell lines and relapse/refractory patient samples to venetoclax. Our studies reveal a potent therapeutic strategy of metabolically driven synthetic lethality involving targeting glutamine metabolism for sensitization to venetoclax in MM.

A functional single-nucleotide polymorphism in the ERCC1 gene alters the efficacy of narrowband ultraviolet B therapy in patients with active vitiligo in a Chinese population.

BACKGROUND: T lymphocytes have been shown to cause the destruction of melanocytes in vitiligo pathogenesis. Narrowband ultraviolet B (NB-UVB), as an effective therapeutic strategy in vitiligo, can lead to the formation of DNA photoproducts such as cyclobutane pyrimidine dimers (CPDs) in perilesional lymphocytes and thus induce skin immunosuppression. The repair of DNA photoproducts is performed mainly through the nucleotide excision repair (NER) pathway. We hypothesized that single-nucleotide polymorphisms (SNPs) in NER genes might influence the repair capacity of CPDs and thus contribute to variations in phototherapy efficiency. OBJECTIVES: To detect genetic polymorphisms in NER genes and their relationship with the efficacy of NB-UVB therapy in patients with active vitiligo. METHODS: We investigated the association of NER SNPs (XPA A23G, XPC Ci11A, XPC C2919A and ERCC1 C118T) with phototherapy efficacy in 86 patients with vitiligo who received NB-UVB treatment. Furthermore, we examined the impact of ERCC1 C118T on the apoptosis of T lymphocytes and CPD accumulation after NB-UVB irradiation. RESULTS: We found that patients with vitiligo with the ERCC1 codon 118 CC genotype showed better efficacy after NB-UVB irradiation than those with the ERCC1 118 TT and CT genotypes, whereas no such association was documented among the genotypes of XPA A23G, XPC Ci11A or XPC C2919A. Additionally, the apoptosis rates and CPD levels of lymphocytes after NB-UVB irradiation in patients with the ERCC1 118 CC genotype were significantly higher than those in patients with the ERCC1 118 TT and CT genotypes. CONCLUSIONS: The ERCC1 118 CC genotype confers better efficacy of NB-UVB therapy in patients with active vitiligo.

Comparative Study of Three Traditional Mongolian Medicines on Streptozotocin-induced Diabetic Nephropathy in Rats.

3 kinds of prescription of Traditional Mongolian (Chinese) Medicine (TMM) have been used in treating diabetic nephropathy (DN). We aimed to investigate: first, which prescription was more effective; second, whether it was more effective when combined with the 3 prescriptions. The DN model was prepared by a single dose of Streptozotocin (STZ, 65 mg/kg, i.p.) in rats and treated 3 times every day with P-1 (Sugmul-10), P-2 (Narenmandul-11), P-3 (Xieriga-4) respectively, and combined group was treated with P-1 in the morning, P-2 and P-3 in the evening. The results showed combining with 3 prescriptions in one day was much more effective than each single prescription. The mechanism of renal protection maybe related to MMP-2 and TGF-ß1, the conclusion could be useful and beneficial for clinical medicine.

Anticonvulsant activity of 2-(substituted-imino)thiazolidin-4-ones.

A novel series of 2-(substituted-imino)thiazolidin-4-ones were synthesized and evaluated for anticonvulsant activity using the maximal electroshock seizure (MES) and subcutaneous pentylenetetrazole (Sc-PTZ) assays and their neurotoxicity was measured by the rotarod test. The results of these tests demonstrated that 2-(4-(pentyloxy)phenylimino)thiazolidin-4-one (5d) was the most potent anticonvulsant, with ED50 value of 18.5 mg/kg and 15.3 mg/kg in the MES and Sc-PTZ tests, and protective index (PI=TD50/ED50) values of 10.6 and 12.8 respectively. 5d was much safer than a reference drug Carbamazepine.

Pharmacokinetic study of icariside II after a single dose administration of YiGu capsule in healthy Chinese volunteers.

To investigate the pharmacokinetic characteristics of icariside II after a single oral dose administration of YiGu Capsule in healthy -Chinese volunteers. 8 (4 female) healthy Chinese volunteers were involved and administrated for 15 doses of YiGu Capsule after fasting overnight. 4 ml of blood samples were collected at scheduled time before and after administration. Icariside II in human plasma was separated on a Agela Venusil XBP-C18 column (250 mm×4.6 mm, 5 µm), and eluted using acetonitrile-water (containing 0.1% formic acid) (70:30, V/V) as mobile phase. The analytes were detected with a -triple quad HPLC-MS/MS using ESI with positive ionization. Ions monitored in the multiple reaction monitoring mode were m/z 515.1 (precursor ion) to m/z 369.1 (product ion) for icariside II and m/z 321.0 (precursor ion) to m/z 275.0 (product ion) for lorazepam (internal standard).The plasma concentration of icariside II was determined by established HPLC-MS/MS method after disposition and its pharmacokinetic parameters were analyzed and evaluated by PK Solver Software (version 1.0). The Cmax, Tmax, t1/2, AUC0-24, AUC0-∞, MRT0-∞-1, 2.50±1.91 h, 7.61±2.81 h, 12.68±4.86 ng · mL-1 · h, 14.22±5.75 ng · mL-1 · h, 9.55±1.56 h, 12.81±7.94 L · h-1 and 121.70±32.70 L. The established HPLC-MS/MS method was sensitive, selective and rapid for icariside II pharmacokinetic study.

The effect of St John's Wort on the pharmacodynamic response of clopidogrel in hyporesponsive volunteers and patients: increased platelet inhibition by enhancement of CYP3A4 metabolic activity.

Clopidogrel is metabolically activated by cytochrome P450 (CYP) isoenzymes. We evaluated whether St. John's wort (SJW), a CYP2C19 and CYP3A4 inducer, enhances the pharmacodynamic response of clopidogrel. Volunteers (n = 45) were screened for clopidogrel hyporesponsiveness after a 300-mg load. After a 7-day washout, hyporesponders (n = 10) received 14 days of SJW (300 mg 3 times a day) followed by a second 300-mg clopidogrel. Platelet aggregation was measured at 0, 2, 4, and 6 hours postloading; hepatic CYP3A4 activity was simultaneously determined at 0 and 4 hours by the erythromycin breath test. A prospective, randomized, double-blind pilot study was conducted in postcoronary stent patients (n = 85) on clopidogrel 75 mg/d screened for clopidogrel hyporesponsiveness. Hyporesponders (n = 20) were randomized to SJW (n = 10) or placebo (n = 10); platelet aggregation was measured before and after 14 days of therapy. In volunteers, SJW decreased platelet aggregation (59% ± 14% vs. 40% ± 15% at 2 hours, P = 0.02; 56% ± 10% vs. 44% ± 13% at 4 hours, P < 0.03; and 55% ± 14% vs. 37% ± 14% at 6 hours, P = 0.01) and increased CYP3A4 activity (2.1% ± 0.4% CO2 exhaled per hour before vs. 2.9% ± 0.6% CO2 exhaled per hour after SJW, P = 0.002). In patients, SJW decreased platelet reactivity (226 ± 39 vs. 185 ± 49 P2Y12 reactivity units, P = 0.0002) and increased platelet inhibition (23% ± 11% vs. 41% ± 16%, P = 0.002). SJW may be a future therapeutic option to increase CYP metabolic activity and antiplatelet effect of clopidogrel in hyporesponders.

Directional differentiation of chicken spermatogonial stem cells in vitro.

BACKGROUND: Mammalian spermatogonial stem cells (SSC) are able to differentiate into different cell types in vitro, which are valuable sources for regenerative medicine and gene transfer studies. We investigated the differentiation potential of chicken SSC into osteoblasts, neuron-like cells and adipocytes in vitro. METHODS: Chicken SSC from the testes of 18- and 20-day-old chicken embryos were cultured in different induction media for three passages in vitro. For differentiation into osteoblasts, SSC were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 1 x 10(-4) micromol/mL desamethasone, 10 micromol/mL (beta-sodium glycerophosphate and 0.05 mg/mL vitamin C, and examined by microscopy after Von Kossa's, cytochemical and immunohistochemical staining. For differentiation into neuron-like cells, SSC were cultured in DMEM supplemented with 1 x 10(-3) micromol/mL retinoic acid (RA), 5.0 micromol/mL 3-isobutyl-1-methylxanthine (IBMX) and examined by microscopy after toluidine blue or immunohistochemical staining. For differentiation into adipocytes, SSC were cultured in DMEM supplemented with 1 x 10(-3) micromol/mL dexamethasone, 0.01 mg/mL insulin, 0.5 micromol/mL IBMX and examined by microscopy after Oil red O staining and reverse transcriptase-polymerase chain reaction (RT-PCR) for gene expression of peroxisome proliferation activation receptor-gamma (PPAR-gamma). RESULTS: After 15 and 21 days of culture in the induction medium for osteoblast differentiation, 75% and 80% chicken SSC differentiated into osteoblasts, as confirmed by Von Kossa's, calcium-cobalt and collagen I antibody staining. After 3 and 7 days of culture in the induction medium for neuron-like cell differentiation, 78% and 85% SSC became neuron-like cells, as confirmed by staining with toluidine blue and the monoclonal antibody against neuron-specific enolase, nestin and glial fibrillary acidic protein. After 7 days of culture in the induction for adipocyte differentiation, 85% SSC differentiated into adipocytes, as confirmed by Oil red O staining and RT-PCT for PPAR-gamma gene expression. DISCUSSION: Our results show that chicken SSC can differentiate into osteoblasts, neuron-like cells and adipocytes under similar conditions as for directional differentiation of mammalian SSC in vitro. The findings show the feasibility of using SSC-derived cells for developmental biology and gene transfer studies in chickens.