[Electroacupuncture improves motor function of rats with osteoarthritis by alleviating joint inflammation through the Wnt-7B/ß-catenin signaling pathway].

OBJECTIVE: To investigate the effect of electroacupuncture on osteoarthritis in rats and explore the possible mechanism. METHODS: Thirty SD rats were randomly divided into osteoarthritis model group, electro-acupuncture group and control group (n=10), and in the former two groups, early osteoarthritis was induced using a modified DMM surgical modeling method. After successful modeling, the rats in the electro-acupuncture group were treated with electro-acupuncture at bilateral "Housanli" and "Anterior knee point". Behavioral tests of the rats were performed and scored using the LequesneMG scale. Subchondral bone degeneration was observed in each group, and serum levels of IL-1ß, ADAMTS-7, MMP-3 and COMP were measured using ELISA. The mRNA and protein expressions of IL-1ß, Wnt-7B, ß-catenin, ADAMTS-7, and MMP-3 in the cartilage tissue of the knee joints were detected using RT-PCR and Western blotting. RESULTS: In behavioral tests, the rats in the model and electroacupuncture groups had significantly higher LequesneMG scores after modeling than those in the control group (P < 0.05). After 20 days of treatment, LequesneMG scores were significantly lowered in rats in the electroacupuncture as compared with the model rats (P < 0.05). Imaging examination revealed obvious subchondral bone damage in both the electroacupuncture group and the model group, but the damages were significantly milder with former group. Compared with the model rats, the rats receiving electroacupuncture had significantly lower serum levels of IL-1ß, ADAMTS-7, MMP-3 and COMP (P < 0.05) with also lower expressions of IL-1ß, Wnt-7B, ß-catenin, ADAMTS-7 and MMP-3 in the cartilage tissues at both the mRNA and protein levels (P < 0.05). CONCLUSION: Electroacupuncture can alleviate joint pain and improve subchondral bone damage in rats with osteoarthritis by reducing IL-1ß levels in the joint cartilage tissue and serum to alleviate joint inflammation and by reducing such cytokines as ADAMTS-7 and MMP-3 via regulating the Wnt-7B/ß-catenin signaling pathway.

[The prospects of DNA damage repair variants guiding platinum compounds in the treatment of triple negative breast cancer].

Triple negative breast cancer (TNBC) is prone to recurrence and metastasis, which is the subtype of poorest prognosis. Chemotherapy is the main treatment, although there is lack of effective adjuvant chemotherapy regimens. The unsatisfactory efficacy of chemotherapy has been a bottleneck in improving the outcome of TNBC. Platinum compounds act directly on DNA to kill tumor cells, and they have a stronger killing effect on tumor cells carrying DNA damage repair (DDR) defects, which is an important entry point to improve the efficacy of TNBC. Biomarkers for predicting the efficacy of platinum drugs in TNBC treatment have always been a hot topic. The DDR pathway contains a large number of related genes, and recent studies have shown that deficiencies in the DDR pathway may be associated with the efficacy of platinum drugs, which is expected to be a biomarker for predicting the efficacy of platinum drugs in breast cancer treatment.

[Chinese traditional five-tone music therapy].

The ideology of Chinese traditional five-tone music therapy started in the five-tone music practice for witch medicine (Wu Yi) or Shaman medicine (Sa Man Yi Xue) treatment in the primitive period. It came from the five-element thinking, i.e, wood, fire, earth, metal and water which matches each inner organ in the human body to diagnose the lesion of the five main organs in traditional Chinese medicine. The five-tone system diagnoses the lesion of the five inner organs based on the changes of the five tones Jue, Zheng, Gong, Shang and Yu, and the construction of the traditional Chinese medicine theories. It eventually formed in the Yellow Emperor's Internal Classic (Huang Di Nei Jing)during the Warring Period. Chinese traditional five-tone music therapy evolved and developed in each historical dynasty, and laid an ideological and theoretical foundation for contemporary Chinese traditional five-tone music therapy. It helped construct a music therapy as a traditional cultural treasure with unique Chinese cultural characteristics.

Supplementation with Yucca schidigera improves antioxidant capability and immune function and decreases fecal score of dairy calves before weaning.

Yucca schidigera (YS) is a species of plant rich in antimicrobials, antioxidants, and immunomodulators. It has been used as feed additive to improve animal performance and decrease methane emissions in cattle. However, few studies have evaluated YS in dairy calves. In this study, we evaluated the effects of YS on the growth performance, antioxidant capacity, and immune function in dairy calves before weaning. We randomly assigned 40 newborn female Holstein calves (4 d old; 40 ± 5 kg of body weight) to 1 of 4 treatments (n = 10 per treatment), which were fed 0, 3, 6, or 9 g/d of YS powder. The YS allowance was mixed into milk or milk replacer and fed twice daily. Dry matter intake (both liquid and starter feed) and fecal score were recorded daily, and body weight, withers height, body length, and heart girth were measured at 4, 14, 28, 42, and 60 d of age. Blood was sampled from the jugular vein at 14, 42, and 60 d of age after the afternoon feeding for analysis of serum antioxidant capacity and immune function. Feeding YS did not affect dry matter intake, but decreased the feed-to-gain ratio with a quadratic dose effect. Over the whole study period, the average daily gain tended to linearly increase with the increasing YS doses, and it was 6.8% higher in diets supplemented with 9 g/d of YS than in the basal control diet without YS. The YS supplementation linearly decreased fecal score in a dose-dependent manner, and the frequency of diarrhea was significantly decreased as the YS supplementation increased throughout the whole study period. The YS supplementation also linearly decreased maleic dialdehyde concentration in the serum compared with the control group. The activity of catalase tended to linearly and quadratically increase, and that of glutathione peroxidase increased linearly with the increased YS supplementation. Serum concentrations of IgA and IgG increased linearly with the increased YS supplementation, and that of IgG tended to increase quadratically. To the best of our knowledge, this is the first study that demonstrated that feeding YS to young calves could improve growth, feed efficiency, and immunity, and decrease fecal score and diarrhea. The results of this study indicated that feeding YS at 9 g/d may be recommended to benefit dairy calves before weaning.

Should postoperative haemoglobin and potassium levels be checked routinely following blood-conserving primary total joint arthroplasty?

AIMS: Despite declining frequency of blood transfusion and electrolyte supplementation following total joint arthroplasty, postoperative blood analyses are still routinely ordered for these patients. This study aimed to determine the rate of blood transfusion and electrolyte restoration in arthroplasty patients treated with a perioperative blood conservation protocol and to identify risk factors that would predict the need for transfusion and electrolyte supplementation. PATIENTS AND METHODS: Patients undergoing primary total joint arthroplasty of the hip or knee between July 2016 and February 2017 at a single institution were included in the study. Standard preoperative and postoperative laboratory data were collected and reviewed retrospectively. A uniform blood conservation programme was implemented for all patients. Need for blood transfusion or potassium supplementation was determined through a coordinated decision by the care team. Rates of transfusion and supplementation were observed, and patient risk factors were noted. RESULTS: The overall rate of blood transfusion was 1.06% in the study population of 1132 total joint arthroplasties performed in 1023 patients. Of the 12 patients requiring transfusion, 11 were female, ten occurred in patients undergoing total hip arthroplasty, and all 12 patients had a preoperative haemoglobin level less than 130 g/l. Operative duration and surgical blood loss were significantly greater in those patients requiring blood transfusion. Nearly all patients requiring transfusion had a history of, or risk factors for, cardiovascular disease. Potassium supplementation was required in 15.5% of the study cohort; 72% of these patients receiving potassium presented with a potassium level less than 4 mmol/l during preoperative testing, while the remaining 28% had a past medical history of either significant anaemia, cardiopulmonary, cardiovascular, or renal diseases that had required substantial medical management. CONCLUSION: A consistent blood-conserving perioperative strategy effectively minimized need for blood transfusion in total joint arthroplasty patients below previously reported rates in the literature. We suggest that postoperative full blood counts and basic metabolic panels should not routinely be ordered in these patients unless their preoperative haemoglobin and potassium is below 130 g/dl or 4 mmol/l respectively, and they have medical comorbidities.

Effects of source on bioavailability of selenium, antioxidant status, and performance in lactating dairy cows during oxidative stress-inducing conditions.

In the current study, we used heat stress (HS) as an oxidative stress model to examine the effects of hydroxy-selenomethionine (HMSeBA), an organic selenium source, on selenium's bioavailability, antioxidant status, and performance when fed to dairy cows. Eight mid-lactation Holstein dairy cows (141 ± 27 d in milk, 35.3 ± 2.8 kg of milk/d, parity 2 or 3) were individually housed in environmental chambers and randomly assigned to 1 of 2 treatments: inorganic Se supplementation (sodium selenite; SS; 0.3 mg of Se/kg of dry matter; n = 4) or HMSeBA supplementation (0.3 mg of Se/kg of dry matter; n = 4). The trial was divided into 3 continuous periods: a covariate period (9 d), a thermal neutral (TN) period (28 d), and a HS period (9 d). During the covariate and TN periods, all cows were housed in TN conditions (20°C, 55% humidity). During HS, all cows were exposed to cyclical HS conditions (32-36°C, 40% humidity). All cows were fed SS during the covariate period, and dietary treatments were implemented during the TN and HS periods. During HS, cows fed HMSeBA had increased Se concentrations in serum and milk, and total Se milk-to-serum concentration ratio compared with SS controls. Superoxide dismutase activity did not differ between Se sources, but we noted a treatment by day interaction in glutathione peroxidase activity as HS progressively reduced it in SS controls, whereas it was maintained in HMSeBA cows. Supplementation with HMSeBA increased total antioxidant capacity and decreased malondialdehyde, hydrogen peroxide, and nitric oxide serum concentrations compared with SS-fed controls. We found no treatment effects on rectal temperature, respiratory rate, or dry matter intake. Supplementing HMSeBA tended to increase milk yield and decrease milk fat percentage. No other milk composition parameters differed between treatments. We observed no treatment effects detected on blood biochemistry, except for a lower alanine aminotransferase activity in HMSeBA-fed cows. These results demonstrate that HMSeBA supplementation decreases some parameters of HS-induced oxidative stress.

Rehmanniae Radix Preparata suppresses bone loss and increases bone strength through interfering with canonical Wnt/ß-catenin signaling pathway in OVX rats.

Rehmanniae Radix Preparata (RRP) improves bone quality in OVX rats through the regulation of bone homeostasis via increasing osteoblastogenesis and decreasing osteoclastogenesis, suggesting it has a potential for the development of new anti-osteoporotic drugs. INTRODUCTION: Determine the anti-osteoporotic effect of RRP in ovariectomized (OVX) rats and identify the signaling pathway involved in this process. METHODS: OVX rats were treated with RRP aqueous extract for 14 weeks. The serum levels of tartrate-resistant acid phosphatase (TRAP), receptor activator of nuclear factor kappa-Β ligand (RANKL), alkaline phosphatase (ALP), and osteoprotegerin (OPG) were determined by ELISA. Bone histopathological alterations were evaluated by H&E, Alizarin red S, and Safranin O staining. Bone mineral density (BMD) and bone microstructure in rat femurs and lumbar bones were determined by dual-energy X-ray absorptiometry and micro-computed tomography. Femoral bone strength was detected by a three-point bending assay. The expression of Phospho-glycogen synthase kinase 3 beta (p-GSK-3ß), GSK-3ß, Dickkopf-related protein 1 (DKK1), cathepsin K, OPG, RANKL, IGF-1, Runx2, ß-catenin, and p-ß-catenin was determined by western blot and/or immunohistochemical staining. RESULTS: Treatment of OVX rats with RRP aqueous extract rebuilt bone homeostasis demonstrated by increasing the levels of OPG as well as decreasing the levels of TRAP, RANKL, and ALP in serum. Furthermore, RRP treatment preserved BMD and mechanical strength by increasing cortical bone thickness and epiphyseal thickness as well as improving trabecular distribution in the femurs of OVX rats. In addition, RRP downregulated the expression of DKK1, sclerostin, RANKL, cathepsin K, and the ratio of p-ß-catenin to ß-catenin, along with upregulating the expression of IGF-1, ß-catenin, and Runx2 and the ratio of p-GSK-3ß to GSK-3ß in the tibias and femurs of OVX rats. Echinacoside, jionoside A1/A2, acetoside, isoacetoside, jionoside B1, and jionoside B2 were identified in the RRP aqueous extract. CONCLUSION: RRP attenuates bone loss and improves bone quality in OVX rats partly through its regulation of the canonical Wnt/ß-catenin signaling pathway, suggesting that RRP has the potential to provide a new source of anti-osteoporotic drugs.

Longitudinal Association between Selenium Levels and Hypertension in a Rural Elderly Chinese Cohort.

OBJECTIVES: Results from previous studies have been inconsistent on the association between selenium and hypertension, and very few studies on this subject have focused on the elderly population. The purpose of this study is to examine the relationship between selenium level and hypertension in a rural elderly Chinese cohort. DESIGN: A longitudinal study was implemented and data were analyzed using logistic regression models and Cox proportional hazards regression model adjusting for potential confounders. The associations between selenium level and prevalent hypertension at baseline and between selenium and incident hypertension were examined. SETTING: Community-based setting in four rural areas in China. SUBJECTS: A total of 2000 elderly aged 65 years and over (mean 71.9±5.6 years) participated in this study. MEASUREMENTS: Nail selenium levels were measured in all subjects at baseline. Blood pressure measures and self-reported hypertension history were collected at baseline, 2.5 years and 7 years later. Hypertension was defined as systolic blood pressure 140 mmHg or higher, diastolic blood pressure 90 mmHg or higher, or reported use of anti-hypertensive medication. RESULTS: The rate of baseline hypertension was 63.50% in this cohort and the mean nail selenium level is 0.413±0.183µg/g. Multi-covariate adjusted cross-sectional analyses indicated that higher selenium level was associated with higher blood pressure measures at baseline and higher rates of hypertension. For the 635 participants with normal blood pressure at baseline, 360 had developed hypertension during follow-up. The incidence rate for hypertension was 45.83%, 52.27%, 62.50%, 70.48%, and 62.79% from the first selenium quintile to the fifth quintile respectively. Comparing to the lowest quintile group, the hazard ratios were 1.41 (95%CI: 1.03 to1.94), 1.93 (95%CI: 1.40 to 2.67), 2.35 (95%CI: 1.69 to 3.26) and 1.94 (95%CI: 1.36 to 22.77) for the second selenium quintile to the fifth quintile respectively. CONCLUSIONS: Our findings suggest that high selenium may play a harmful role in the development of hypertension. Future studies are needed to confirm our findings and to elucidate a plausible biological mechanism.

Effectiveness of rubber seed oil and flaxseed oil to enhance the α-linolenic acid content in milk from dairy cows.

This experiment was conducted to investigate effect of rubber seed oil compared with flaxseed oil when fed alone or in combination on milk yield, milk composition, and α-linolenic acid (ALA) concentration in milk of dairy cows. Forty-eight mid-lactation Holstein dairy cows were randomly assigned to 1 of 4 treatments according to a completely randomized design. Cows were fed a basal diet (control; CON) or a basal diet supplemented with 4% rubber seed oil (RO), 4% flaxseed oil (FO), or 2% rubber seed oil plus 2% flaxseed oil (RFO) on a dry matter basis for 9 wk. Feed intake, milk protein percentage, and milk fat levels did not differ between the treatments. Cows fed the RO, FO, or RFO treatments had a higher milk yield than the CON group (up to 10.5% more), whereas milk fat percentages decreased. Compared with the CON, milk concentration of ALA was substantially higher in cows receiving RO or RFO, and was doubled in cows receiving FO. The ALA yield (g/d) increased by 31.0, 70.3, and 33.4% in milk from cows fed RO, FO, or RFO, respectively, compared with the CON. Both C18:1 trans-11 (vaccenic acid) and C18:2 cis-9,trans-11 (conjugated linoleic acid; CLA) levels were higher in cows fed added flaxseed or rubber seed oil. The CLA yield (g/d) increased by 336, 492, and 484% in cows fed RO, FO, or RFO, respectively, compared with the CON. The increase in vaccenic acid, ALA, and CLA was greater in cows fed RFO than in cows fed RO alone. Compared with the CON, the milk fat from cows fed any of the dietary supplements had a higher concentration of unsaturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids; conversely, the saturated fatty acids levels in milk fat were 30.5% lower. Insulin and growth hormones were not affected by dietary treatments; however, we noted an increase in both cholesterol and nonesterified fatty acids levels in the RO, FO, or RFO treatments. These results indicate that rubber seed oil and flaxseed oil will increase milk production and the concentration of functional fatty acids (ALA, vaccenic acid, and CLA) in milk fat while decreasing the content of saturated fatty acids.

Contact fatigue of human enamel: Experiments, mechanisms and modeling.

Cyclic contact between natural tooth structure and engineered ceramics is increasingly common. Fatigue of the enamel due to cyclic contact is rarely considered. The objectives of this investigation were to evaluate the fatigue behavior of human enamel by cyclic contact, and to assess the extent of damage over clinically relevant conditions. Cyclic contact experiments were conducted using the crowns of caries-free molars obtained from young donors. The cuspal locations were polished flat and subjected to cyclic contact with a spherical indenter of alumina at 2Hz. The progression of damage was monitored through the evolution in contact displacement, changes in the contact hysteresis and characteristics of the fracture pattern. The contact fatigue life diagram exhibited a decrease in cycles to failure with increasing cyclic load magnitude. Two distinct trends were identified, which corresponded to the development and propagation of a combination of cylindrical and radial cracks. Under contact loads of less than 400N, enamel rod decussation resisted the growth of subsurface cracks. However, at greater loads the damage progressed rapidly and accelerated fatigue failure. Overall, cyclic contact between ceramic appliances and natural tooth structure causes fatigue of the enamel. The extent of damage is dependent on the magnitude of cyclic stress and the ability of the decussation to arrest the fatigue damage.

Genipin is active via modulating monoaminergic transmission and levels of brain-derived neurotrophic factor (BDNF) in rat model of depression.

Genipin, an important bioactive component from Gardenia jasminoides Eills, was demonstrated to possess antidepressant-like effects in a previous study. However, the molecular mechanism of antidepressant-like effects on genipin was not clear. The present study aimed to investigate the possible mechanism of antidepressant-like effects on genipin with a chronic unpredictable mild stress (CUMS)-induced depression model in rats. In CUMS-induced depressive rats, bodyweight and 1% sucrose consumption decreased significantly compared with the normal control group. Furthermore, these changes could be significantly reversed by genipin application. The levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE) in the hippocampus decreased and the level of 5-hydroxyindole acetic acid (5-HIAA) increased in the CUMS-induced depressive rats. However, pre-treatments with genipin significantly increased the levels of 5-HT, NE and decreased the level of 5-HIAA in the hippocampus. The concentration of cAMP in the hippocampus was increased by genipin compared to the CUMS-exposed model group. The mRNA expressions of 5-hydroxytryptamine 1A receptor (5-HT1AR), cAMP response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in rats were decreased exposed to CUMS, which were reversed by genipin-treated rats exposed to CUMS. Compared to the CUMS-exposed model group, the mRNA expression of 5-hydroxytryptamine 2A receptor (5-HT(2A)R) was decreased significantly by genipin-treated rats. The mRNA and protein expression of CREB, BDNF were increased in genipin-treated rats compared to the CUMS-exposed model group. Moreover, the levels of corticosterone in serum were decreased by genipin-treated compared to the CUMS-exposed model group. These results suggest that the possible mechanism of antidepressant-like effects on genipin, at least in one part, resulted from monoaminergic neurotransmitter system and the potential dysfunctional regulation of the post-receptor signaling pathway, which particularly affected the 5-HT(1A)R, 5-HT(2A)R and BDNF levels in the hippocampus.

Effects of linseed oil and palm oil on growth performance, tibia fatty acid and biomarkers of bone metabolism in broilers.

1. This study was conducted to determine the effects of different dietary fat sources on growth performance, tibia fatty acids and biomarkers of bone metabolism in broilers. 2. One-d-old commercial Arbor Acres broilers were fed with a maize-soya bean basal diet for 42 d, supplemented with oils according to the following 5 treatments: lard (lard group); linseed oil (linseed oil group); palm oil (palm oil group); linseed oil + palm oil (60:40 or 40:60 w/w, LP-1 group and LP-2 group, respectively). 3. No significant differences in weight gain, feed intake and gain/feed ratio were observed between the lard and linseed oil groups. Birds fed on palm oil had significantly greater weight gain and feed intake than those fed on lard or linseed oil. Growth performance in LP-1 and LP-2 was significantly greater than that of single-oil groups. 4. Tibia growth and bone characteristics were not influenced by supplementation with lard, linseed oil, or palm oil alone, but broilers fed on a mixture of fats had significantly greater tibia weight and length compared to broilers fed on linseed oil. Bone mineral density in tibia was significantly increased in LP-1 and LP-2 groups. 5. Supplementation of linseed oil alone or in combination with palm oil enhanced apparent digestibility of calcium, reduced serum calcium and increased tibia calcium concentrations. Moreover, supplementation with linseed oil alone or in combination with palm oil had a positive effect on biomarkers of bone growth. 6. The combination of linseed and palm oils was beneficial for growth performance, tibia growth and biomarkers of bone metabolism.

Vegetation changes and partitioning of selenium in 4-year-old constructed wetlands treating agricultural drainage.

The knowledge of selenium (Se) partitioning in treatment wetlands and wetland vegetation management are essential for long-term effective operation of constructed wetlands treating Se-laden agricultural tile-drainage in central California. In this field study, samples from different compartments of treatment wetlands were collected and the vegetation change in each wetland cell was examined four years after the wetland's inception. The results showed that saltgrass (Distichlis spicata) and rabbitfoot grass (Polypogon monspeliensis) were less competitive than cattail (Typha latifolia) and saltmarsh bulrush (Scirpus robustus). Over 90% of the wetland cell originally vegetated with saltgrass or rabbitfoot grass was occupied by invasive plants--i.e., when invasive species were not controlled in the wetlands. More Se was likely found in sediments from vegetated regions, compared to the unvegetated areas of the wetland cell. Particularly, rhizosphere sediments accumulated about 4-fold more Se than non-rhizosphere sediments. Among the total Se retained in the wetland 90% of the total Se was partitioned in the top 10-cm layer of sediment. The Se accumulation in plant materials accounted for about 2% of the total Se mass retained in each wetland cell. This field study demonstrated that wetland plants play significant roles in the treatment of Se-laden agricultural drainage.

[Construction of root library by SSH and preliminary analysis of genes responsible for phosphorus deficiency in maize].

An elite maize inbred line with high tolerance to low phosphorus, 178, was studied for constructing root library and analyzing some genes closely related to phosphorus (P) deficiency using SSH and Semi-quantitative RT-PCR. As a result, 3648 preliminary clones were obtained for root library under stress of P deficiency. By DNA sequencing of 34 random clones, we obtained 23 unique EST sequences which are involved in functions of root cell structure, tolerance and defense, protein modification and composition, transcription regulation, metabolism, and other unknown aspects. Five representative genes were further analyzed for their expression models. The results suggested that the molecular mechanism to adapt P deficiency in maize, performed by multi-genes with different contributions, is similar to rice, Arabidopsis and soybean. The expression order of 5 low P tolerant genes in maize root was PAP, GCS, TOM, PDI and AIP. And it was considered preliminarily that physiological and biochemical changes were prior to morphologic changes in maize root and the essential tolerance to low P may be determined by extending absorption of P to wide soil range through adaption of root architecture and root secretions, which is the greatest difference between tolerant and sensitive maize varieties under low P stress.

Induction of apoptosis in HepG2 cells by solanine and Bcl-2 protein.

The nightshade (Solanum nigrum Linn.) has been widely used in Chinese traditional medicine as a remedy for the treatment of digestive system cancer. The anti-tumor activity of solanine, a steroid alkaloid isolated from the nightshade has been demonstrated. To observe the effect of anti-tumor and mechanism of solanine. The MTT assay was used to evaluate the IC(50) on the three digestive system tumor cell lines. The effect on the morphology was observed with a laser confocal microscopy; the rate of apoptosis and the cell cycle were measured using flow cytometry (FCM); the expression of Bcl-2 protein was measured by Western blot. The results show that the IC(50) for HepG(2), SGC-7901, and LS-174 were 14.47, >50, and >50 microg/ml, respectively; the morphology of cells in the negative control was normal; for the treated groups, typical signs for apoptosis were found. The rate of apoptosis in HepG(2) cells induced by solanine was found to be 6.0, 14.4, 17.3, 18.9, and 32.2%, respectively. Observation of the cell cycle showed that cells in the G(2)/M phases disappeared while the number of cells in the S phase increased significantly for treated groups. Western blot showed that solanine decreased the expression of Bcl-2 protein. Therefore, the target of solanine in inducing apoptosis in HepG(2) cells seems to be mediated by the inhibition in the expression of Bcl-2 protein.

Chemical status of selenium in evaporation basins for disposal of agricultural drainage.

Evaporation basins (or ponds) are the most commonly used facilities for disposal of selenium-laden saline agricultural drainage in the closed hydrologic basin portion of the San Joaquin Valley, California. However concerns remain for potential risk from selenium (Se) toxicity to water fowl in these evaporation basins. In this study, we examined the chemical status of Se in both waters and sediments in two currently operating evaporation pond facilities in the Tulare Lake Drainage District. Some of the saline ponds have been colonized by brine-shrimp (Artemia), which have been harvested since 2001. We evaluated Se concentration and speciation, including selenate [Se(VI)], selenite [Se(IV)], and organic Se [org-Se or Se(-II)] in waters and sediment extracts, and fractionation (soluble, adsorbed, organic matter (OM)-associated, and Se(0) and other resistant forms) in sediments and organic-rich surface detrital layers from the decay of algal blooms. Selenium in ponds without vascular plants exhibited similar behavior to wetlands with vascular plant present, indicating that similar Se transformation processes and mechanisms had resulted in Se immobilization and an increase of reduced Se species [Se(IV), org-Se, and Se(0)] from Se(VI)-dominated input waters. Selenium concentrations in most pond waters were significantly lower than the influent drainage water. This decrease of dissolved Se concentration was accompanied by the increase of reduced Se species. Selenium accumulated preferentially in sediments of the initial pond cell receiving drainage water. Brine-shrimp harvesting activities did not affect Se speciation but may have reduced Se accumulation in surface detrital and sediments.

Arsenic speciation and accumulation in evapoconcentrating waters of agricultural evaporation basins.

To sustain agricultural productivity, evaporation basins (or ponds) have been widely used for the disposal of agricultural drainage in areas requiring subsurface drainage in the San Joaquin Valley of California, USA. The drainage water contains elevated concentration of trace elements including selenium (Se) and arsenic (As). Unlike Se, little information is available about As, a potentially high risk element. The objective of this study was to characterize the chemical behavior of As and acquire data for better understanding of biogeochemical processes and conditions affecting As fate in evaporation ponds. The study site was a 726 ha evaporation basin facility (containing 10 cells with water flowing in series) in the hydrologically closed Tulare Basin of California. We examined water chemistry, As concentration and speciation along the water flow path between cells as well as within the cells. Arsenic concentrations in the water increased linearly with Cl(-), a conservative ion from evapoconcentration. Reduced As species as arsenite [As(III)] and organic arsenic (org-As) also increased with increases in Cl(-) and salinity. Water samples with elevated EC (i.e., towards the end of flow path) had high dissolved organic matter, low dissolved oxygen, and elevated sulfide concentrations, indicating the development of reducing conditions. We hypothesize that such changes could facilitate the reduction of arsenate [As(V)] to As(III) and org-As. Elevated As in sediment profiles indicate a solid phase sink mechanism, but not significant enough to remove and reduce As concentrations in the water columns. These findings help us better define the processes that affect As in drainage facilities and contribute to our understanding of how As behaves in other regions of the world that have similar climatic and hydrogeochemical conditions.

Selenium removal and mass balance in a constructed flow-through wetland system.

A field study on the removal of Se from agricultural subsurface drainage was conducted from May 1997 to February 2001 in the Tulare Lake Drainage District (TLDD) of San Joaquin Valley, California. A flow-through wetland system was constructed consisting of ten 15- x 76-m unlined cells that were continuously flooded and planted with either a monotype or combination of plants, including sturdy bulrush [Schoenoplectus robustus (Pursh) M.T. Strong], baltic rush (Juncus balticus Willd.), smooth cordgrass (Spartina alterniflora Loisel.), rabbitsfoot grass [Polypogon monspeliensis (L.) Desf.], salt-grass lDistichlis spicata (L.) Greene], cattail (Typha latifolia L.), tule [Schoenoplectus acutus (Muhl. ex Bigelow) A. Löve & D. Löve], and widgeon grass (Ruppia maritima L.). One cell had no vegetation planted. The objectives of this research were to evaluate Se removal efficiency of each wetland cell and to carry out a mass balance on Se. The inflow drainage water to the cells had average annual Se concentrations of 19 to 22 microg L(-1) dominated by selenate [Se(VI), 95%]. Average weekly water residence time varied from about 3 to 15 d for Cells 1 through 7 (target 7 d), 19 to 33 d for Cells 8 and 9 (target 21 d), and 13 to 18 d for Cell 10 (target 14 d). Average weekly Se concentration ratios of outflow to inflow ranged from 0.45 to 0.79 and mass ratio (concentration x water volume) from 0.24 to 0.52 for year 2000, that is, 21 to 55% reduction in Se concentration and 48 to 76% Se removal in mass by the wetland, respectively. The nonvegetated cell showed the least Se removal both in concentration and in mass. The global mass balance showed that on the average about 59% of the total inflow Se was retained within the cells and Se outputs were outflow (35%), seepage (4%), and volatilization (2%). Independent measurements of the Se retained in the cells totaled 53% of the total Se inflow: 33% in the surface (0-20 cm) sediment, 18% in the organic detrital layer above the sediment, 2% in the fallen litter, < 1% in the standing plants, and < 1% in the surface water. Thus, about 6% of the total Se inflow was unaccounted for in the internal compartments.

Cloning and expression of a single-chain catalytic antibody that acts as a glutathione peroxidase mimic with high catalytic efficiency.

Glutathione peroxidase (GPX) has a powerful role in scavenging reactive oxygen species. In previous papers we have developed a new strategy for generating abzymes: the monoclonal antibody with a substrate-binding site is first prepared, then a catalytic group is incorporated into the monoclonal antibody's binding site by using chemical mutation [Luo, Zhu, Ding, Gao, Sun, Liu, Yang and Shen (1994) Biochem. Biophys. Res. Commun. 198, 1240-1247; Ding, Liu, Zhu, Luo, Zhao and Ni (1998) Biochem. J. 332, 251-255]. Since then we have established a series of catalytic antibodies capable of catalysing the decomposition of hydroperoxides by GSH. The monoclonal antibody 2F3 was raised against GSH-S-2,4-dinitrophenyl t-butyl ester and exhibited high catalytic efficiency, exceeding that of rabbit liver GPX, after chemical mutation. To produce pharmaceutical proteins and to study the reason why it exhibits high catalytic efficiency, we sequenced, cloned and expressed the variable regions of 2F3 antibody as a single-chain Fv fragment (2F3-scFv) in different bacterial strains. The amounts of 2F3-scFv proteins expressed from JM109 (DE3), BL21 (DE3), and BL21 (coden plus) were 5-10%, 15-20% and 25-30% of total bacterial proteins respectively. The 2F3-scFv was expressed as inclusion bodies, purified in the presence of 8 M urea by Co(2+)-immobilized metal-affinity chromatography (IMAC) and renatured to the active form in vitro by gel filtration. The binding constants of the active 2F3-scFv for GSH and GSSG were 2.46 x 10(5) M(-1) and 1.03 x 10(5) M(-1) respectively, which were less by one order of magnitude than that of the intact 2F3 antibody. The active 2F3-scFv was converted into selenium-containing 2F3-scFv (Se-2F3-scFv) by chemical modification of the reactive serine; the GPX activity of the Se-2F3-scFv was 3394 units/micromol, which approaches the activity of rabbit liver GPX.