An investigation into the mechanism of nobiletin's inhibition of papillary thyroid cancer using network pharmacology analysis and experimental pharmacology.

OBJECTIVE: Surgery and radioactive iodine therapy are the main treatments for papillary thyroid carcinoma (PTC), and effective drugs are lacking. As a promising natural product, nobiletin (NOB) has a wealth of pharmacological activities like anti-tumor, antivirus, and other effects. In this research, bioinformatics methods and cellular assays were combined to explore how NOB inhibited PTC. MATERIALS AND METHODS: Our NOB targets were derived from three databases, including the SwissTargetPrediction database, Traditional Chinese Medicine System Pharmacology Database, and the TargetNet server. Four databases were used to identify disease-related targets: GeneCards, PharmGkb, Online Mendelian Inheritance in Man, and DisGeNET. Finally, cross-targets of disease and drug were deemed as pharmacological targets, and they were used for GO and KEGG enrichment analysis. STRING and Cytoscape were applied for PPI Network and core Targets Ranking. Molecular docking analysis validated binding affinity values for NOB and core targets. By using cell proliferation and migration assays, NOB was assessed for its effects on PTC proliferation and migration phenotype. Western blot validated the downregulation of the PI3K/Akt pathway. RESULTS: (1) Preliminarily, 85 NOB targets were predicted for NOB intervention in PTC. (2) Our core target screening identified TNF, TP53, and EGFR, and our molecular docking results confirmed good binding between NOB and protein receptors. (3) NOB inhibited proliferation and migration of PTC cells. PI3K/AKT pathway target proteins were downregulated. CONCLUSIONS: (1) Bioinformatics analyses revealed that NOB may inhibit PTC by regulating TNF, TP53, EGFR and PI3K/AKT signalling pathway. (2) As evidenced by cell experiments, there was an inhibition of proliferating and migrating PTCs by NOB via the PI3K/AKT signalling pathway.

[Effects of the composite of buckwheat-oat-pea on blood glucose in diabetic rats].

OBJECTIVE: To study the effects of buckwheat-oat-pea (BOP) composite flour [buckwheat ∶ oats ∶ peas=6 ∶ 1 ∶ 1 (quality ratio)] on blood glucose in diabetic rats. METHODS: In this study, 64 male Sprague-Dawley rats were divided into 8 groups by fasting blood glucose (FBG) and body weight: normal control group, model control group, metformin group, buckwheat group, oats group, BOP low-dose group (BOP-L), medium-dose group (BOP-M), and high-dose group (BOP-H). The rats in the normal control group were fed with normal diet, the rats in the model control group and metformin group were fed with a high-fat diet (HFD), and the rats in the buckwheat group, oats group, and BOP-L, BOP-M, BOP-H groups were fed with HFD containing 10% buckwheat flour, 10% oat flour, 3.3% BOP, 10% BOP, 30% BOP, respectively. The HFD in all the groups had the same percentage of energy from fat (45%). After 30 days, the rats fed with HFD received intraperitoneal injection of streptozotocin (30 mg/kg, once a week for two weeks) to establish diabetes mellitus. After the model was successful established, the rats were fed for another 28 days. During the study, the body weight, food intake/body weight (FI/BW) and water intake/body weight (WI/BW), food utilization rate, 24 h urine volume, FBG, glucose area under curve (GAUC) of oral glucose tolerance test were measured regularly. At the end of the study, the fasting serum glucose and insulin were measured, and homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. RESULTS: With the inducing of HFD and streptozotocin, compared with the normal control group, the rats in the model control group had higher FI/BW, WI/BW, 24 h urine volume, FBG, GAUC, HOMA-IR (P < 0.05), and lower body weight, food utilization rate (P < 0.05). Compared with the model control group, the rats in the three BOP groups all had higher body weight, food utilization rate (P < 0.05), and lower WI/BW, HOMA-IR (P < 0.05); the rats in the BOP-L and BOP-M groups had lower FI/BW, 24 h urine volume, FBG (P < 0.05), and the rats in the BOP-M group also had lower GAUC (P < 0.05). After the establishment of diabetes, there was no significant difference in blood glucose and the other indicators between the rats in the three BOP groups and the buckwheat group or the oats group (P>0.05). CONCLUSION: The BOP had the effects of reducing blood glucose, insulin resistance and diabetic symptoms on diabetic rats, and had the value for further development and utilization.

The efficacy and safety of docetaxel, cisplatin and fluorouracil (TPF)-based induction chemotherapy followed by concurrent chemoradiotherapy for locoregionally advanced nasopharyngeal carcinoma: a meta-analysis.

OBJECTIVES: In recent years, docetaxel, cisplatin and fluorouracil (TPF)-based induction chemotherapy (IC) has been widely applied in the treatment of locoregionally advanced nasopharyngeal carcinoma (LA-NPC). However, it remains unclear whether TPF is the ideal IC regimen. Thus, we carried out a meta-analysis to compare the efficacy and safety of TPF-based IC plus concurrent chemoradiotherapy (CCRT) versus CCRT alone or double-drug-based IC plus CCRT for LA-NPC. METHODS: We systematically searched PubMed, Embase and the Cochrane Library from inception until December 2018. After rigorous screening of all relevant studies that reported the use of TPF-based IC followed by CCRT for patients with LA-NPC, eight studies met the inclusion criteria and were assessed for design and quality. Among them, three articles were classified as having a high risk of bias and were excluded from the meta-analysis. The outcomes, including overall survival (OS), progression-free survival (PFS), distant metastasis-free survival (DMFS), locoregional failure-free survival (LRFFS) and incidence of adverse events, were pooled with the use of hazard ratio (HR) or odds ratio (OR). Heterogeneity and sensitivity analyses were also carried out. RESULTS: Five trials involving 4223 patients were included in the meta-analysis. Compared to CCRT alone, TPF-based IC plus CCRT significantly improved OS (HR 0.54, 95% confidence interval [CI] 0.35-0.84, P = 0.006), PFS (HR 0.64, 95% CI 0.46-0.88, P = 0.006), LRFFS (HR 0.57, 95% CI 0.34-0.94, P = 0.03), and DMFS (HR 0.58, 95% CI 0.38-0.88, P = 0.01). Moreover, compared to double-drug-based IC plus CCRT, OS (HR 0.74, 95% CI 0.62-0.87, P = 0.0004), PFS (HR 0.76, 95% CI 0.66-0.88, P = 0.0001) and LRFFS (HR 0.75, 95% CI 0.61-0.92, P = 0.006) were also significantly improved by TPF-based IC plus CCRT. Notably, TPF-based IC plus CCRT mainly led to an increased risk of hematologic toxicities, such as leucopenia (OR = 3.20, 95% CI 2.13-4.81, P < 0.0001) and neutropenia (OR = 3.84, 95% CI 0.66-22.36, P = 0.13). However, these were uncomplicated and manageable with growth factor support. CONCLUSIONS: Compared to CCRT alone or double-drug-based IC plus CCRT, TPF-based IC plus CCRT results in better survival outcomes with manageable toxicities. Thus, it is reasonable to recommend the addition of TPF-based IC to CCRT as an excellent choice for patients with LA-NPC.

Epigallocatechin-3-gallate local pre-exposure application prevents SHIV rectal infection of macaques.

Epigallocatechin-3-gallate (EGCG), a natural and major ingredient of green tea, has been shown to have anti-inflammation and anti-HIV-1 properties. We demonstrated that the intrarectal administration of EGCG could protect rhesus macaques from repetitive, intrarectal challenges with low-dose SHIVSF162P3N. This protection has a per-exposure risk reduction of 91.5% (P = 0.0009; log-rank test) and a complete protection of 87.5% (P < 0.001; Fisher's exact test). All protected animals showed no evidence of systemic and mucosal SHIV infection as demonstrated by the absence of viral RNA, DNA and antibodies. In contrast, all controls became infected after repeated SHIV challenges (a median of 2.5 times, range of 1-8 times). Mechanistically, EGCG could block the binding of HIV-1 gp120 to CD4 receptor and suppress the macrophage infiltration/activation in the rectal mucosa of macaques. These data support further clinical evaluation and development of EGCG as a novel, safe and cost-effective microbicide for preventing sexual transmission of HIV-1.

First Report of Anthracnose of Malva sylvestris Caused by Colletotrichum trifolii in China.

Common mallow (Malva sylvestris L.) is a perennial medicinal plant in the Malvaceae family, which is native to Asia, Europe, and North Africa. In July 2012, typical symptoms of anthracnose disease, with a disease incidence of ~70%, were observed on common mallow in the Medicinal Herb Garden of Shenyang Pharmaceutical University, Liaoning, China. The fungus mainly infected the stalks and leaves of M. sylvestris. Pinpoint, brownish lesions initially appeared at the flowering stage and the disease spread within the field. The lesions on stems gradually enlarged and became dark brown, elliptical, and slightly concave. Concurrently, acervuli and mucilaginous conidial masses of the pathogen appeared on lesions under moist conditions. Conidia were hyaline, one-celled, cylindrical with both ends rounded, and measured 10.0 to 12.5 × 2.5 to 4.0 µm (mean 11.3 × 3.3 µm). The fungus was isolated from symptomatic tissues. Small pieces from leaves and stems were surface disinfested with 70% ethanol and 1.5% sodium hypochlorite for 1 min, then rinsed three times with sterile distilled water, and cultured on potato dextrose agar (PDA) at 25°C. The colonies on PDA had initially white aerial mycelia, and later became greenish black with regularly whorled rings. To confirm Koch's postulates, five 3-month-old plants of M. sylvestris were inoculated with a conidial suspension (105 conidia/ml) prepared from PDA cultures incubated for 14 days. Five non-inoculated plants served as controls. The plants were maintained in the greenhouse at 22 to 25°C and about 75% relative humidity under natural daylight. Typical symptoms on inoculated plants were reproduced after ~10 to 14 days, whereas control plants remained asymptomatic. The pathogen was successfully recovered from symptomatic tissues and re-identified, completing Koch's postulates. The internal transcribed spacer (ITS) and large subunit -28S (LSU) region of rDNA was amplified with primers ITS1/ITS4 and NL1/NL4, respectively, and sequenced. Phylogenetic trees (ITS and LSU) that were obtained using MEGE3.1 with the neighbor-joining method showed that both of the isolates fall in the Colletotrichum trifolii clade. The representative sequences (ITS and LSU) were deposited in GenBank (Accession Nos. KJ155692 and KJ920935). The fungus isolated from symptomatic tissues was identified as C. trifolii on the basis of morphological, cultural characteristics, and sequence analysis (2). According to previous references, C. orbiculare and C. malvarum on Malvaceae were respectively described in America and Europe (1,3,4). However, the isolate from M. sylvestris significantly differed from those of C. orbiculare and C. malvarum in cultural characteristics and sequence analysis. In this paper, the results showed that M. sylvestris is a new host of C. trifolii. To our knowledge, this is the first report of mallow anthracnose caused by C. trifolii in China. References: (1) J. A. Bailey et al. Phytopathology 86:1076, 1996. (2) U. Damm et al. Fungal Divers. 61:29, 2013. (3) K. Hyde et al. Fungal Divers. 39:147, 2009. (4) L. Tosi et al. Plant Dis. 88:425, 2004.

Treatment of hydrogen molecule abates oxidative stress and alleviates bone loss induced by modeled microgravity in rats.

UNLABELLED: Treatment with molecular hydrogen alleviates microgravity-induced bone loss through abating oxidative stress, restoring osteoblastic differentiation, and suppressing osteoclast differentiation and osteoclastogenesis. INTRODUCTION: Recently, it has been suggested that hydrogen gas exerts a therapeutic antioxidant activity by selectively reducing cytotoxic reactive oxygen species (ROS). The aim of the present study was to elucidate whether treatment with molecular hydrogen alleviated bone loss induced by modeled microgravity in rats. METHODS: Hindlimb suspension (HLS) and rotary wall vessel bioreactor were used to model microgravity in vivo and in vitro, respectively. Sprague-Dawley rats were exposed to HLS for 6 weeks to induced bone loss and simultaneously administrated with hydrogen water (HW). Then, we investigated the effects of incubation with hydrogen-rich medium (HRM) on MC3T3-E1 and RAW264.7 cells exposed to modeled microgravity. RESULTS: Treatment with HW alleviated HLS-induced reduction of bone mineral density, ultimate load, stiffness, and energy in femur and lumbar vertebra. Treatment with HW alleviated HLS-induced augmentation of malondialdehyde content and peroxynitrite content and reduction of total sulfhydryl content in femur and lumbar vertebra. In cultured MC3T3-E1 cells, incubation with HRM inhibited modeled microgravity-induced ROS formation, reduction of osteoblastic differentiation, increase of ratio of receptor activator of nuclear factor kappa B ligand to osteoprotegerin, inducible nitric oxide synthetase upregulation, and Erk1/2 phosphorylation. In cultured RAW264.7, incubation with HRM aggravated modeled microgravity-induced ROS formation, osteoclastic differentiation, and osteoclastogenesis. CONCLUSION: Treatment with molecular hydrogen alleviates microgravity-induced bone loss in rats. Molecular hydrogen could thus be envisaged as a nutritional countermeasure for spaceflight but remains to be tested in humans.

First Report of Sclerotinia Rot of Chinese Atractylodes Caused by Sclerotinia nivalis in China.

Chinese atractylodes (Atractylodes japonica Koidz.ez Kitam.) is a perennial herb in the Compositae family, and is widely distributed in China. The dried rhizomes of the plant are used in traditional Chinese medicine. During the summer of 2011, typical signs and symptoms of Sclerotinia rot were observed on Chinese atractylodes in a production field of Liaoning Province of China. Symptoms were observed in plants at the flowering stage, distributed in patches throughout the rows, and with a disease incidence of approximately 10 to 15%. The lower mature leaves of infected plants first became yellow and wilted, basal stem areas showed a black-brown rot at the same time under conditions of high humidity, and white cottony mycelium formed along the basal stem and soil surfaces. Ultimately, the basal stem and roots rotted and the plants wilted and died quickly. Black, irregular sclerotia (average 0.8 to 6.9 mm in diameter) were also observed within the pith cavity of split stems and rotted roots. The pathogen was isolated from symptomatic tissues and sclerotia, surface disinfested with 2% sodium hypochlorite, and cultured on potato dextrose agar (PDA) (1). The fungus was mesophilic, with an optimum temperature for mycelial growth in culture of about 20°C. Colonies on PDA produced masses of white aerial mycelium, with small white flocci distributed among sclerotia. After 2 weeks, sclerotia 0.5 to 4.5 mm in diameter were produced near the margin in a uniform distribution. Sclerotia were spherical, elongated, or fused to form irregular shapes and tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. DNA sequences of five replicates were obtained using the TianGen DNA secure plant kit. The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and sequenced. BLAST analysis of the 513-bp segment showed high similarity (99%) with a sequence of Sclerotinia nivalis (GenBank Accession No. AB516670). A representative sequence was deposited in GenBank (Accession No. JX294862). The fungus isolated from symptomatic tissues was identified as S. nivalis Saito on the basis of morphological and cultural characteristics (2,3) and ITS sequence analysis. Symptoms were reproduced in the greenhouse by inoculating the basal stem and roots of 15 atractylodes plants at the 7- to 10-leaf stage. Inoculum was prepared by macerating 14-day-old PDA cultures of the fungus in a blender and placing the mixture (approximately 20 g) into the potting medium of each plant. Sterile PDA was used to inoculate the five control plants. Plants were maintained in a greenhouse at 22 to 25°C and about 75% relative humidity. After 7 to 10 days, symptoms were similar to those in the fields. Lower leaves of inoculated plants became yellow and wilted, and infected plants died 2 weeks after inoculation, whereas control plants remained healthy. The pathogen was successfully recovered from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of Sclerotinia rot of Chinese atractylodes. Given its wide host range, S. nivalis has great potential to become an economically important plant pathogen. References: (1) W. G. Kim and W. D. Cho. Mycobiology 30:41, 2002. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience 38:227, 1997.

Sclerotinia Rot of Windflower (Pulsatilla koreana Nakai) Caused by Sclerotinia nivalis in China.

Windflowers (Pulsatilla spp.) are perennial medicinal plants in the family Ranunculaceae with high economic as well as medicinal value in China. It is a commonly used traditional Chinese medicine (1). In 2012, a stem rot disease was observed on windflower (P. koreana Nakai) at flowering stages in fields of Liaoning Province, China. Disease incidence ranged from 10 to 65% (average of 45%) and resulted in approximately 25 to 60% yield loss. The infected area of plants initially takes on a dark green or brown water-soaked appearance and then becomes paler. Soon after, plants turn brown or black and die. If moisture conditions remain conducive, a cottony mycelium cover was observed on the affected area. Later, the mycelium usually produces numerous black sclerotia that were up to 1 cm long in affected plant parts. Diseased tissue were surface sterilized for 1 min in 1% NaOCl (v/v), rinsed with sterilized distilled water, and plated on potato dextrose agar (PDA). Isolates were mesophilic, with an optimum growth temperature of around 20°C. Colonies on PDA were white, with abundant aerial mycelium. Some mycelium in the colony center, especially that submerged in the medium, was light brown. Sclerotia were spherical to subspherical, elongated or fused to form irregular shapes, 2.5 to 9.0 × 2.0 to 6.8 mm. They were tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. These characteristics were consistent with Sclerotinia nivalis (2,3). Two isolates were selected for molecular identification. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (4) and sequenced. The obtained sequences (GenBank Accession Nos. JX206828 and JX424615) showed 99 and 100% homology with the sequences of S. nivalis in GenBank (Accession No. AB516670). To demonstrate pathogenicity, mycelial blocks of the isolates grown on PDA were placed on the base of the stems of ten 1-month-old Pulsatilla koreana plants. The same numbers of control plants were treated with PDA plugs as a control. The inoculated plants were incubated at 25°C with a 12-h photoperiod. After 4 days, the initiation of stem necrosis was observed, and 9 days after inoculation, the plants collapsed and died. Control plants had no symptoms. The same fungus was reisolated from all inoculated plants, satisfying Koch's postulates. To our knowledge, this is the first confirmed report of windflower as a natural host for S. nivalis in China. References: (1) S. C. Bang et al. J. Nat. Prod. 68:268, 2005. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience. 38:227, 1997. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Severe Outbreak of Rust Caused by Coleosporium pulsatillae Detected on Pulsatilla spp. in China.

Windflower (Pulsatilla spp.) is a perennial medicinal plant in the Ranunculaceae with high economic value as well as medicinal value in China. It is a commonly used Chinese herbal medicine. In 2007, two species, Pulsatilla koreana Nakai and P. chinensis Regel, were observed with severe rust symptoms at three locations (Shenyang City, Benxi City, and Fushun County) in Liaoning, China. The diseased area was estimated to be more than 500 ha and the yield was reduced by 30% on average with up to 65% yield losses in some fields. Since its first record in 2007, the disease has been recorded every year in parts of China. A survey of all cultivated fields in August 2011 revealed that 90% of the Pulsatilla plants were heavily infected with rust. Early symptoms on the adaxial leaf surfaces were small, circular, yellow spots that later enlarged, coalesced, and developed necrotic centers. On the abaxial side, numerous yellow rust uredinia were visible. Urediniospores are globose or ellipsoidal, sometimes angular in shape, with a yellowish content, and measured 22.6 to 39.4 × 15.2 to 23.9 µm. Spore walls were coarsely verrucose when examined by light and scanning electron microscopy. Telia and teliospores were observed in mid-August. Hypophyllous telia often formed around uredinial clusters. Telia were 0.3 to 1.1 mm wide and erumpent with numerous teliospores in a compact layer. Teliospores were clavate, oblong to ellipsoidal, 60.2 to 120.8 × 12.1 to 24.4 µm, gelatinous, and one celled. Teliospores were four celled with the division of the protoplast into an internal four-celled basidium. Pathogenicity tests included inoculation of young P. koreana plants by spraying a urediniospore suspension (30,000 spores/ml of deionized water). Inoculated plants were incubated at 25°C for 48 h, and typical rust symptoms and uredinia developed in 10 to 15 days. On the basis of symptomatology, the host, and morphology of uredinial and telia, the fungus was identified as Coleosporium pulsatillae (Str.) Lév (2). The internal transcribed spacer (ITS) region was amplified using ITS1-F and ITS4-B primers (3), and an amplified product of 817 bp, specific for the species C. pulsatillae, was obtained. The sequence was deposited in GenBank (Accession No. JQ029765). Although this pathogen has been mentioned as part of a fungal species survey from China, it was not fully described (4). This pathogen has been reported on Anemone chinensis in Austria, Sibiria (2), and Korea (1). It has not been reported from anywhere else in China. To our knowledge, this is the first fully described record and most severe outbreak of C. pulsatillae on windflower. References: (1) W. D. Cho and H. D. Shin. List of Plant Diseases in Korea. Korean Society of Plant Pathology, Seoul, Korea, 2004. (2) J. B. De-Toni. Sylloge Fungorum 7:754, 1888. (3) M. Gardes et al. Mol. Ecol. 2:113, 1993. (4) F. L. Tai. Sylloge Fungorum Sinicorum. Science Press, Beijing, 1979.

The impact of obesity on receipt of adjuvant chemotherapy for breast cancer in the National Comprehensive Cancer Network (NCCN) centers.

Disparities in the receipt of adjuvant chemotherapy for early stage breast cancer is an important factor influencing mortality. We investigated whether greater body mass index (BMI) decreases receipt of adjuvant chemotherapy among women with operable breast cancer. In the NCCN breast cancer outcomes database, we identified women aged ≤ 70 with newly diagnosed stage I, II, or III breast cancer between 1997 and 2007, for whom use of adjuvant chemotherapy was classified as either standard-of-care or discretionary based on their clinical characteristics. Body mass index was assessed in categories (<18.5 kg/m(2) [underweight], 18.5 to <25 kg/m(2) [normal], 25 to <30 kg/m(2) [overweight], 30-39 kg/m(2) [obese], ≥ 40 kg/m(2) [extreme obese]). Multivariable logistic regression analysis was used to examine the association between BMI and receipt of chemotherapy in each classification group. 9,527 women were eligible for the study; 40% normal weight or less; 31% overweight; 24% obese; and 5% extremely obese. In multivariable analysis, there was no significant association between BMI and receipt of chemotherapy in either classification group. Among women for whom chemotherapy would be considered standard-of-care, older age (P < 0.001), comorbidity (P < 0.001), and non-Hispanic black ethnicity (P = 0.002) were associated with a lower likelihood of receipt of chemotherapy; however, the effect of ethnicity was not modified by obesity. Among women treated for operable breast cancer in the NCCN centers, BMI had no impact on receipt of adjuvant chemotherapy and did not modify the lower likelihood of chemotherapy among non-Hispanic black patients. Further investigation is needed into other factors that contribute to patient disparities in the receipt of chemotherapy in major academic centers.

Prenatal ethanol exposure attenuates GABAergic inhibition in basolateral amygdala leading to neuronal hyperexcitability and anxiety-like behavior of adult rat offspring.

Prenatal exposure to a relatively high-dose ethanol (EtOH) caused anxiety-like behavior of adult male rat offspring. Previous studies have demonstrated that GABA system in the basolateral amygdala complex (BLA) is involved in the pathogensis of anxiety-related disorders. The role of GABAergic system in the BLA was investigated in anxiety-like behavior evoked by prenatal EtOH exposure. The infusion of midazolam (MDZ), a positive modulator of GABA(A) receptor, into the BLA prevented anxiety-like behavior in EtOH-offspring without affecting the corresponding behavior of control offspring. The data suggest that anxiety-like behavior could be causally related to increased neuronal excitability attributable to depressed GABAergic inhibition in the BLA. To test this hypothesis, evoked potential was studied using brain slices from EtOH-offspring. Potential evoked in the BLA by single stimuli applied to external capsule showed multispike responses, indicative of GABAergic disinhibition. These multiple responses were no longer evident after the perfusion with MDZ. In the slices from EtOH-offspring, paired-pulse inhibition (GABA(A)-dependent) was suppressed. Also, in EtOH-offspring, long-term potentiation (LTP) was induced by a single train of high frequency stimulation, which did not induce LTP in control rats. Moreover, MDZ pretreatment prevented the facilitating effect of EtOH on LTP induction. The data provide the functional evidence that prenatal EtOH exposure attenuates GABAergic inhibition in the BLA resulting in neuronal hyperexcitability and anxiety-like behavior of adult rat offspring.

Hybrid pulse anodization for the fabrication of porous anodic alumina films from commercial purity (99%) aluminum at room temperature.

Most porous anodic alumina (PAA) or anodic aluminum oxide (AAO) films are fabricated using the potentiostatic method from high-purity (99.999%) aluminum films at a low temperature of approximately 0-10 degrees C to avoid dissolution effects at room temperature (RT). In this study, we have demonstrated the fabrication of PAA film from commercial purity (99%) aluminum at RT using a hybrid pulse technique which combines pulse reverse and pulse voltages for the two-step anodization. The reaction mechanism is investigated by the real-time monitoring of current. A possible mechanism of hybrid pulse anodization is proposed for the formation of pronounced nanoporous film at RT. The structure and morphology of the anodic films were greatly influenced by the duration of anodization and the type of voltage. The best result was obtained by first applying pulse reverse voltage and then pulse voltage. The first pulse reverse anodization step was used to form new small cells and pre-texture concave aluminum as a self-assembled mask while the second pulse anodization step was for the resulting PAA film. The diameter of the nanopores in the arrays could reach 30-60 nm.

Huperzine A for Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) has become a major public health problem around the world due to its increasing prevalence, long duration, caregiver burden, and high financial cost of care. The degeneration of acetylcholine-containing neurons in the basal forebrain has been implicated in the symptoms of AD. Cholinesterase inhibitors may block the degradation of acetylcholine, thus increasing the efficacy of the remaining cholinergic neurons. Huperzine A is a linearly competitive, reversible inhibitor of acetyl cholinesterase that is said to have both central and peripheral activity with the ability to protect cells against hydrogen peroxide, beta-amyloid protein (or peptide), glutamate, ischemia and staurosporine-induced cytotoxicity and apoptosis. These properties might qualify Huperzine A as a promising agent for treating dementia (including AD). OBJECTIVES: To assess the efficacy and safety of Huperzine A for the treatment of patients with AD. SEARCH STRATEGY: The Specialized Register of the Cochrane Dementia and Cognitive Improvement Group was searched on 1 February 2006 using the search term: huperzin*. The CDCIG Specialized register contains records from all major health care databases (MEDLINE, EMBASE, PsycINFO, CINAHL, SIGLE, ISTP, INSIDE, LILACS) as well as from many trials databases and grey literature sources. In addition, the CBM and AMED databases and relevant websites were searched and some journals were hand-searched. Specialists in the field were approached for unpublished material and any publications found were searched for additional references. SELECTION CRITERIA: All relevant randomized controlled trials (RCTs) studying the efficacy and safety of Huperzine A for AD. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers using a self-developed data extraction form and entered into RevMan 4.2.10 software. Meta-analyses were performed when more than one trial provided data on a comparable outcome on sufficiently similar patients. Random effects analyses were performed whenever heterogeneity between results appeared to be present. Standardized differences in mean outcome measures were used due to the use of different scales and periods of treatment. MAIN RESULTS: Six trials including a total of 454 patients met our inclusion criteria. The methodological quality of most included trials was not high. It was shown that compared to placebo, Huperzine A had beneficial effects on the improvement of general cognitive function measured by MMSE (WMD 2.81; 95% CI 1.87 to 3.76; P < 0.00001) and ADAS-Cog at six weeks (WMD 1.91; 95% CI 1.27 to 2.55) and at 12 weeks (WMD 2.51; 95% CI 1.74 to 3.28), global clinical assessment measured by CDR (WMD -0.80; 95% CI -0.95 to -0.65) and CIBIC-plus (OR 4.32, 95% CI 2.37 to 7.90), behavioral disturbance measured by ADAS-non-Cog at six weeks (WMD -1.33, 95%CI -2.12 to -0.54) and at 12 weeks (WMD -1.52, 95% CI-2.39 to -0.65), and functional performance measured by ADL (WMD = -7.17; 95% CI -9.13 to -5.22; P < 0.00001). However, Huperzine A was not superior to placebo in the improvement of general cognitive function measured by Hasegawa Dementia Scale (HDS) (WMD: 2.78; 95% CI -0.17 to 5.73, P = 0.06) and specific cognitive function measured by Weshler Memory Scale (WMS) (WMD = 6.64; 95% CI -3.22 to 16.50; P = 0.19). No data were available on quality of life and caregiver burden. The adverse events of Huperzine A were mild and there were no significant differences of adverse events between Huperzine A groups and control groups. AUTHORS' CONCLUSIONS: From the available evidence, Huperzine A seems to have some beneficial effects on improvement of general cognitive function, global clinical status, behavioral disturbance and functional performance, with no obvious serious adverse events for patients with AD. However, only one study was of adequate quality and size. There is therefore inadequate evidence to make any recommendation about its use. Rigorous design, randomized, multi-centre, large-sample trials of Huperzine A for AD are needed to further assess the effects.

A novel assay for discovery and characterization of pro-apoptotic drugs and for monitoring apoptosis in patient sera.

We have developed an apoptosis assay based on measurement of a neoepitope of cytokeratin-18 (CK18-Asp396) exposed after caspase-cleavage and detected by the monoclonal antibody M30. The total amount of caspase-cleaved CK18 which has accumulated in cells and tissue culture media during apoptosis is measured by ELISA. The sensitivity is sufficient for use in the 96-well format to allow high-through-put screening of drug libraries. We here describe strategies allowing classification of pro-apoptotic compounds according to their profiles of induction of apoptosis in the presence of pharmacological inhibitors. The time course of induction of CK18 cleavage can furthermore be used to distinguish structurally similar compounds. We propose that compounds that induce rapid CK18 cleavage have mechanisms of actions distinct from conventional genotoxic and microtubuli-targeting agents, and we present one example of an agent that induces almost immediate mitochondrial depolarization and cytochrome c release. Finally, CK18-Asp396 cleavage products are released from cells in tissue culture, and presumably from tumor cells in vivo. These products can be measured in sera from cancer patients. We present evidence suggesting that it will be possible to use the M30-ELISA assay for measuring chemotherapy-induced apoptosis in patient sera, opening possibilities for monitoring therapy.

[Pharmacognostical identification of Polygonum perfoliatum].

In the paper, phamacognostical identification and UV-identification of Polygonum perfoliatum L. were studied.

[Identification of radix rubiae and the root of Humulus scardens (Lour.) Merr].

The paper reported the differences of the Radix Rubiae and the root of Humulus scardens (Lour.) Merr. on morphological and histological characteristics and UV-identification.

Binding of the maize cytosolic Hsp70 to calmodulin, and identification of calmodulin-binding site in Hsp70.

Using a gel-overlay technique of biotinylated calmodulin (CaM), we showed that maize cytosolic Hsp70 protein could bind to CaM in the presence of 1 mM CaCl2. The purified maize cytosolic Hsp70 inhibited the activity of CaM-dependent NADK in a concentration-dependent manner. A synthetic peptide, which possesses the 21 amino acid sequence, PRALRRLRTACERAKRTLSST, at positions 261-281 in maize cytosolic Hsp70, could associate with CaM in the presence of 1 mM calcium. The synthetic peptide inhibited CaM-dependent NADK activity and PDE activity. This indicates that the 21-amino acid sequence at positions 261-281 is the CaM-binding site. The binding of CaM to Hsp70 inhibited the ATPase activity of Hsp70. The possible regulator function of Hsp70 in cell signaling events in response to heat stress is discussed.

Intracellular acidification of human melanoma xenografts by the respiratory inhibitor m-iodobenzylguanidine plus hyperglycemia: a 31P magnetic resonance spectroscopy study.

In vivo 31P magnetic resonance spectroscopy demonstrates that human melanoma xenografts can be significantly acidified by induction of hyperglycemia combined with administration of m-iodobenzylguanidine (MIBG), an inhibitor of mitochondrial respiration. In melanoma xenografts (< or =8 mm diameter), intracellular pH (pHi, measured by the chemical shift of the Pi resonance) and extracellular pH (pHe, measured with 3-aminopropylphosphonate) was reduced by less than 0.2 unit during i.v. infusion of glucose for 40 min. Administration of MIBG (30 mg/kg) under hyperglycemic conditions (26 mM) reduced tumor pHi and pHe by approximately 0.4 (P < 0.001) and approximately 0.6 (P < 0.001) unit, respectively; coincidentally, the nucleoside triphosphates:Pi ratio decreased approximately 60% (P < 0.004) relative to the baseline level. Minimal changes in pHi and pHe and a small decrease in nucleoside triphosphates:Pi ratio (26%, P = 0.2) were observed in liver in response to MIBG plus hyperglycemia. These results suggest that under normoglycemic and hyperglycemic conditions, small human melanoma xenografts (< or =8 mm) may exhibit a relatively high level of oxidative phosphorylation that may be blocked by MIBG. The acidification may result from increased lactate production as a direct effect of MIBG inhibition of respiration in mitochondria of tumor cells, or through indirect systemic effects, which remain to be identified. The synergetic effects of MIBG and hyperglycemia result in significant acidification of the tumor and a decrease in tumor bioenergetic status, and the effects are largely selective for tumors in comparison with normal tissues.

Effects of soaking temperature and soaking time during preparation of water extract of tea on anticlastogenicity against environmental tobacco smoke in the sister-chromatid exchange assay.

Water extract of tea (WET) was prepared by soaking green tea at different temperatures for various periods of time and was used to test whether the soaking temperature and soaking time during the preparation of WET influence the content of polyphenols and the anticlastogenicity of WET against environmental tobacco smoke (ETS). Five major polyphenols in WET were measured. Extractable-respirable particulate (ERP) was obtained from ETS-contaminated indoor air (ERP-ETS). The sister-chromatid exchange assay (SCE) was utilized to evaluate the clastogenic effects of ERP-ETS and the anticlastogenic effects of WET. The results indicate that ERP-ETS is clastogenic and WET has significant anticlastogenic effects on ERP-ETS. The content of polyphenols and the anticlastogenic potential of WET depended on the soaking temperature and soaking time during WET preparation. At the soaking temperature of 80 degrees C, an increased soaking time was correlated with a higher percentage of polyphenols and a concomitantly enhanced anticlastogenic efficacy. By contrast, at the soaking temperature of 100 degrees C, a longer soaking time was associated with a higher percentage of polyphenols concomitant with a lower anticlastogenic efficacy. The data suggest that, besides polyphenols, and additional material(s), which may be partially inactivated at 100 degrees C, is contributing to the anticlastogenicity of WET.

Molecular cloning and sequencing of the sodB gene from a heterocystous cyanobacterium Anabaena sp. PCC 7120.

Superoxide dismutase (Sod) plays an important role in all aerobic organisms. The sodB gene of a heterocystous cyanobacterium Anabaena sp. PCC 7120 was cloned and sequenced. The Sod protein is predicted to have 199 amino acids and a molecular mass of 22.5 kDa. Sequence comparison among SodB from cyanobacteria and chloroplasts revealed that the sodB gene indeed encodes an iron-Sod. Northern blot analysis showed that the sodB gene of Anabaena sp. PCC 7120 is transcribed as a single gene and its expression was up-regulated when the cells were subjected to a shift from a nitrogen repletion condition to a nitrogen depletion condition.