Assessment of genetic diversity and variation of Robinia pseudoacacia seeds induced by short-term spaceflight based on two molecular marker systems and morphological traits.

The black locust (Robinia pseudoacacia) is a forest legume that is highly valued as a honey plant and for its wood. We explored the effect of short-term spaceflight on development of R. pseudoacacia seedlings derived from seeds that endured a 15-day flight; the genetic diversity and variation of plants sampled from space-mutagenized seeds were compared to plants from parallel ground-based control seeds using molecular markers and morphological traits. In the morphology analysis, the space-mutagenized group had apparent variation compared with the control group in morphological traits, including plant height, basal diameter, number of branches, branch stipular thorn length, branch stipular thorn middle width, leaflet vertex angle, and tippy leaf vertex angle. Simple sequence repeat (SSR) and sequence-related amplified polymorphism (SRAP) molecular marker analyses showed a slightly higher levels of genetic diversity in the space-mutagenized group compared to the control group. In the SRAP analysis, the space-mutagenized group had 115 polymorphic bands vs 98 in the controls; 91.27% polymorphic loci vs 77.78% in the controls; 1.9127 ± 0.2834 alleles vs 1.7778 ± 0.4174 in the controls; Nei's genetic diversity (h) was 0.2930 ± 0.1631 vs 0.2688 ± 0.1862 in the controls, and the Shannon's information index (I) was 0.4452 ± 0.2177 vs 0.4031 ± 0.2596 in the controls. The number of alleles was significantly higher in the space-mutagenized group. In the SSR analysis, the space-mutagenized group also had more polymorphic bands (51 vs 46), a greater percentage of polymorphic loci (89.47% vs 80.70%); h was also higher (0.2534 ± 0.1533 vs 0.2240 ± 0.1743), as was I (0.3980 ± 0.2069 vs 0.3501 ± 0.2412). These results demonstrated that the range of genetic variation in the populations of R. pseudoacacia increased after spaceflight. It also suggested that the SSR and SRAP markers are effective markers for studying mutations and genetic diversity in R. pseudoacacia. The data provide valuable molecular evidence for the effects of the space environment on R. pseudoacacia and may contribute to future space-breeding programs involving forest trees.

Effects of total flavonoids and flavonol glycosides from Epimedium koreanum Nakai on the proliferation and differentiation of primary osteoblasts.

In a bioassay-guided drug screening for anti-osteoporosis activity, eight flavonol glycosides were isolated from Epimedium koreanum Nakai, which is traditionally widely used in China for the treatment of impotence and osteoporosis. The effects of total flavonoids and flavonol glycosides on the proliferation and differentiation of rat calvarial osteoblast-like cells were evaluated by the MTT method and measuring the activity of alkaline phosphatase (ALP activity). Total flavonoids (1.2 x10(-2) to 6.0 x10(-7) mg/ml) and flavonol glycosides (2.0 x10(-5) to 1.0 x10(-9) mol/l) exhibited a strong inhibition on the proliferation of primary osteoblasts at most concentrations. However, the total flavonoids and icariin significantly promoted the differentiation of primary osteoblasts. The results suggested that flavonoids from E. koreanum Nakai may improve the development of osteoblasts by promoting the ALP activity; and icariin might be one of the active constituents facilitating the differentiation of osteoblasts.

Reversal of multidrug resistance in cancer cells by Rhizoma Alismatis extract.

Prolonged chemotherapy may lead to the selective proliferation of multidrug resistant (MDR) cancer cells. In MDR HepG2-DR and K562-DR cells that over-expressed P-glycoprotein (Pgp), the extract of the rhizomes of Alisma orientalis (Sam) Juzep. showed a synergistic growth inhibitory effect with cancer drugs that are Pgp substrates including actinomycin D, puromycin, paclitaxel, vinblastine and doxorubicin. At the same toxicity levels the herbal extract was more effective than verapamil, a standard Pgp inhibitor, in enhancing cellular doxorubicin accumulation and preventing the efflux of rhodamin-123 from the MDR cells. The extract restored the effect of vinblastine on the induction of G(2)/M arrest in MDR cells. Our data suggest that A. orientalis may contain components that are effective inhibitors of Pgp.

Involvement of NF-kappaB and c-myc signaling pathways in the apoptosis of HL-60 cells induced by alkaloids of Tripterygium hypoglaucum (levl.) Hutch.

Tripterygium hypoglaucum (levl.) Hutch (Celastraceae) (THH) root is a Chinese medicinal herb commonly used for treating autoimmune diseases. In the present study, alkaloids of THH were prepared and their cytotoxicity against the HL-60 cell was investigated. THH-induced apoptosis was observed using flow cytometry, confocal fluorescence microscope, and DNA laddering and caspase assays. The molecular mechanism involved in the induction of HL-60 cell apoptosis by THH alkaloids was examined using cDNA microarrays containing 3000 human genes derived from a leukocyte cDNA library. Sixteen genes were identified to be differentially expressed in HL-60 cells upon THH treatment. Several genes related to the NF-kappaB signaling pathway and cell apoptosis (such as NFKBIB, PRG1 and B2M) were up-regulated. In addition, c-myc binding protein and apoptosis-related cysteine proteases caspase-3 and caspase-8 were also regulated. The changes in c-Myc RNA expression and c-myc protein level were further confirmed by RT-PCR and Western blot analysis. The results demonstrated that THH alkaloids induced apoptosis of HL-60 cells though c-myc and NF-kappaB signaling pathways.

A new guaianolide as apoptosis inhibitor from Chrysanthemum boreale.

The 8-O-acetyl-3,10-dihydroxy-4(15),11(13)-guaiadien-12,6-olide, named borenolide, was isolated from Chrysanthemum boreale M. Borenolide inhibits the etoposide-induced apoptosis in U937 cell with an IC(50) value of 6.2 microg/ml. Structural assignment was based on NMR-spectroscopic methods and X-ray crystallographic analysis.

Changes in Ca, Cu, Fe, Mg, and Zn contents in mouse brain tissues after prolonged oral ingestion of brick tea liquor containing a high level of Al.

A study was conducted to analyze the regional distribution of Ca, Cu, Fe, Mg, and Zn contents in brain tissues after animals were given liquor of brick tea that contained a high Al content. In 25 normal adult male mice given either water or 0.9% NaCl for 1 mo or 2 mo, the metal concentrations in the serum, liver, frontal cortex, hippocampus, cerebellum, and brain stem were comparable (p > 0.05). When the drinking water was replaced by a 1% brick tea liquor, which contained a high Al content, serum Al concentration was increased significantly 1 mo after the onset of the experiment and remained high at the end of the second month. The level of Al was also elevated in both the cortex and hippocampus at 1 mo after replacing tea for drinking water. In addition to Al, there were a significant increase in hippocampal Zn and a decrease in Cu contents. There was no change in tissue Mg or Fe contents, but there was a significant increase in Ca content in every brain region studied. It was suggested that the increase in Ca might be the result of the effect of other components in tea. Unlike the brain, there was no change in the concentration of any of the metals, including Al, in the liver, which further demonstrated that the changes observed in the brain was specific. The results of the present study confirmed that Al, when given orally in the form of tea, could be absorbed into the bloodstream. The absorbed Al could accumulate in selected brain regions. The presence of Al might also change the tissue content of endogenous trace metals.

Enhanced iron uptake of Saccharomyces cerevisiae by heterologous expression of a tadpole ferritin gene.

We genetically engineered Saccharomyces cerevisiae to express ferritin, a ubiquitous iron storage protein, with the major heavy-chain subunit of tadpole ferritin. A 450-kDa ferritin complex can store up to 4,500 iron atoms in its central cavity. We cloned the tadpole ferritin heavy-chain gene (TFH) into the yeast shuttle vector YEp352 under the control of a hybrid alcohol dehydrogenase II and glyceraldehyde-3-phosphate dehydrogenase promoter. We confirmed transformation and expression by Northern blot analysis of the recombinant yeast, by Western blot analysis using an antibody against Escherichia coli-expressed TFH, and with Prussian blue staining that indicated that the yeast-expressed tadpole ferritin was assembled into a complex that could bind iron. The recombinant yeast was more iron tolerant in that 95% of transformed cells, but none of the recipient strain cells, could form colonies on plates containing 30 mM ferric citrate. The cell-associated concentration of iron was 500 microg per gram (dry cell weight) of the recombinant yeast but was 210 microg per gram (dry cell weight) in the wild type. These findings indicate that the iron-carrying capacity of yeast is improved by heterologous expression of tadpole ferritin and suggests that this approach may help relieve dietary iron deficiencies in domesticated animals by the use of the engineered yeast as a feed and food supplement.

Increased expression of cyclooxygenase-2 protein in human gastric carcinoma.

Gastric adenocarcinoma is one of the most common malignancies in the world, and yet little is known about its molecular process of development and progression. Recent studies have suggested that ingestion of nonsteroid anti-inflammatory drugs reduces the risk of colon cancer, presumably by inhibiting the cyclooxygenase (COX) enzyme. COX-2, one isoform of the COX enzyme, is the rate-limiting enzyme in prostaglandin synthesis, and the function of this enzyme is thought to relate to inflammatory processes and carcinogenesis. To understand the role of COX enzyme in gastric cancer, we measured COX-2 expression in 104 human gastric carcinoma tissues by immunohistochemical analysis. We obtained tissue specimens from 104 surgically resected gastric adenocarcinoma patients. We performed immunohistochemical stain for human COX-2 with polyclonal antibody in gastric carcinoma. After curative resection and extensive lymph node dissection, all patients received adjuvant chemotherapy containing 5-fluorouracil. Expression of COX-2 showed cytoplasmic staining, not only in cancer cells but also in precancerous lesions such as metaplastic and adenomatous cells. We confirmed up-regulation of COX-2 in gastric cancer tissues compared with normal paired mucosa using Western blot analysis. There was no correlation between clinicopathological characteristics of gastric cancer patients and intensity of COX-2 protein expression. This study indicates that COX-2 protein over-expression may contribute to an early event of gastric cancer development, and it further suggests that selective inhibition of COX-2 may provide a chemopreventive effect against gastric carcinogenesis.

Betel quid chewing and risk of adverse pregnancy outcomes among aborigines in southern Taiwan.

It is known that substance use is associated with increased risk of adverse pregnancy, outcomes. The aims of this study were to estimate the prevalence of alcohol, cigarette, betel quid and drug use during pregnancy and to assess the risk of adverse effects of betel quid chewing on pregnancy outcomes in aboriginal women in southern Taiwan. The study population included 62 women with adverse pregnancy outcomes and 124 age-matched women. Subjects were interviewed at their homes by trained interviewers using a structure questionnaire. Prevalences of various substance use in aborigines with adverse pregnancy outcomes were estimated as follows: alcohol, 43.6%; smoking, 14.5%; betel quid chewing, 43.6% and over-the-counter drug use, 8.1%; whereas in the comparison group it was alcohol, 38.7%; smoking, 8.1%; betel quid chewing, 28.2% and none used drugs. Univariate analysis revealed that adverse pregnancy outcomes were associated with maternal betel quid chewing, maternal illness during pregnancy, and the number of pregnancies (gravidity) experienced. After adjusting for maternal illness and number of previous pregnancies covariates, the prevalence of adverse pregnancy outcome was computed to be 2.8-fold higher among betel quid chewing women as compared to non-chewers (AOR=2.8, 95% CI=1.2-6.8). Among the aboriginal women, prenatal care is essential not only for routine care, but also to focus health education on the harmful effects of substance use, especially betel quid use during pregnancy.

PIP: This study aims to estimate the prevalence of alcohol, cigarette, betel quid and drug use during pregnancy. Additionally, this paper also seeks to assess the risks and adverse effects of betel quid chewing on pregnancy outcomes among aboriginal women in southern Taiwan. The study included 62 women with adverse pregnancy outcomes and 124 age-matched women. Subjects were interviewed using a structure questionnaire. Univariate analyses between case group and comparison group by maternal age, education level, occupation, religious affiliation, family structure, gravidity, maternal illness, intervals of pregnancy, blood relations, age of getting married, husband's occupation and husband's drinking and smoking habits were examined. The analysis revealed that adverse pregnancy outcomes were associated with maternal betel liquid chewing, maternal illness during pregnancy, and the number of pregnancies experienced. In aboriginal women, prenatal care is essential not only for routine care, but also to health education concerning the harmful effects of substance use, especially using betel liquid during pregnancy.

Hemistepsins with cytotoxic activity from Hemisteptia lyrata.

Two sesquiterpene lactones were isolated from Hemisteptia lyrata Bunge (Compositae). Their structures were elucidated by spectroscopic methods as recently known hemistepsin and the new 6-O-(2-methylpropenoyl)-3-hydroxy-4(15), 10(14), 11(13)-guaiatrien-12,8-olide. The two hemistepsin derivatives were examined for their in vitro cytotoxic activity against human cell lines such as UACC 62, HCT 15, UO-31, PC-3 and A549. Growth inhibitory property was determined by in vitro treatment of the respective cell lines using the sulforhodamine B assay (SRB). Both compounds exhibited significant cytotoxic activity (IC50 < 4.3 micrograms/ml) on all five human cell lines tested.

Prevalences and related factors of substances use in female aborigines in southern Taiwan.

Alcohol use and betel quid chewing are common in aborigines in Taiwan. The aims of this study were to evaluate the patterns of drinking, smoking, betel quid chewing and to assess the related factors of the above substance use among female aborigines. Three hundred female Bunun aborigines aged between 17 and 50 years were recruited into this study via a stratified-random sampling. Subjects were interviewed at their homes by trained interviewers using a structured questionnaire. Prevalences of various substances use were estimated as follows: alcohol, 67.7%; betel quid chewing, 36.4%; smoking, 11.6%; and over-the-counter drugs, 8%. Related factors of substances use were analyzed by multiple logistic regression as follows. Alcohol use was significantly related to low educational level, where the spouse was a drinker and there was an attitude in favor of female drinking. Significant factors related to smoking were folk religion and where the spouse was a smoker. Significant factors related to betel chewing were low educational level, and where the spouse was a betel chewer, and poor knowledge about the adverse health effects of betel chewing. We also found women who consumed alcohol and chewed betel quid were significantly to show more external health locus control. About 80% of women recognized the adverse effects of alcohol consumption and smoking to pregnancy outcomes. Although they knew the harmful effects of substance use, only half of them stopped using these substances during their pregnancy. Health education on the harmful effects of alcohol, betel quid, and cigarettes, especially during pregnancy is essential in primary care.

Prevalence and related risk factors of betel quid chewing by adolescent students in southern Taiwan.

The prevalence and related risk factors of betel quid chewing among adolescent students were studied in a junior high school (group 1) and in a vocational school (group 2) in southern Taiwan. Group 1 consisted of 3548 participants (89.7% response rate) and group 2 of 1358 (97.6% response rate). The students were asked to complete a questionnaire anonymously. In the junior high school 1.9% of students including all grades (13-15 years old) and both sexes was found to be a current betel quid chewer and 14% was an ex-chewer, whereas 10.2% of vocational school students (16-18 years old) was a current chewer and 31% was an ex-chewer. The prevalence of betel chewing was significantly higher among boys than girls. A high proportion of chewers was also a smoker and alcohol drinker. A statistical analysis of sociodemographic factors showed that male students who smoked tobacco, consumed alcohol and were friends or classmates of students who were betel quid chewers, were the likeliest adolescents to chew betel quid.

Induction of metallothionein in the Reuber H-35 rat hepatoma cell.

The feasibility of using the Reuber H-35 rat hepatoma cell (RH-35 cells) as model for studying metallothionein induction was examined. The RH-35 cells were treated with Cd, a toxic metal which is known to induce metallothionein. The LC50 after a 3-h treatment was 70 microM. The value was significantly higher (P < 0.05) if the cells were pre-treated with a sublethal dose of CdCl2 (5 microM) for 2 days, indicating that pre-treatment with a low dose of Cd can protect against a subsequent higher dose of the same metal. Both the mRNA and the gene product metallothionein can be identified in the cells 2 days after treatment with 5 microM Cd. In addition to Cd, Zn and Cu were also able to induce the expression of metallothionein to various degrees. The results indicate that the MT gene is present in RH-35 cells and is responsive to treatment with various metals. Thus, this cell line can be used as a model to study metallothionein induction.

Aluminum deposition in the central nervous system. Preferential accumulation in the hippocampus in weanling rats.

Simultaneous administration of 1,25-dihydroxyvitamin-D3, citrate, and aluminum-containing phosphate binders is frequently used in patients with chronic renal failure. In order to investigate whether citrate may represent a risk factor of aluminum intoxication, 16 Sprague-Dawley weanling rats were randomly assigned to four groups: 1,25-dihydroxyvitamin-D3 at 16 ng/kg/day was given to all groups except the control; in addition, two groups received either aluminum hydroxide at 160 mg elemental aluminum/kg/day, or aluminum citrate at 160 mg elemental aluminum/kg/day, respectively. The control group received only the vehicle. Extremely high aluminum concentrations were detected in the hippocampus of rats receiving aluminum compounds. This content of aluminum (microgram/g dry weight) was far higher than that found in other brain areas of the same animals (146.40 +/- 51.23 versus 4.49 +/- 0.62, P less than 0.001) as well as that detected in the hippocampus of the control animals (2.73 +/- 0.40). Thus, in non-uremic, weanling rats supplemented with 1,25-dihydroxyvitamin-D3, the administration of aluminum favors selective accumulation in the hippocampus. No differences between aluminum hydroxide and aluminum citrate administration were observed.