Transcriptome analysis reveals corresponding genes and key pathways involved in heat stress in Hu sheep.

Heat stress (HS) seriously affects animal performance. In view of global warming, it is essential to understand the regulatory mechanisms by which animals adapt to heat stress. In this study, our aim was to explore the genes and pathways involved in heat stress in sheep. To this end, we used transcriptome analysis to understand the molecular responses to heat stress and thereby identify means to protect sheep from heat shock. To obtain an overview of the effects of heat stress on sheep, we used the hypothalamus for transcriptome sequencing and identified differentially expressed genes (DEGs; false discovery rate (FDR) < 0.01; fold change > 2) during heat stress. A total of 1423 DEGs (1122 upregulated and 301 downregulated) were identified and classified into Gene Ontology (GO) categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Heat stress triggered dramatic and complex alterations in gene expression in the hypothalamus. We hypothesized that heat stress induced apoptosis and dysfunction in cells and vital organs and affected growth, development, reproduction, and circadian entrainment via the calcium signaling pathway, which influences ribosome assembly and function. Real-time PCR was used to evaluate the expression of the genes regulating important biological functions or whose expression profiles were significantly changed after acute heat stress (FDR < 0.01; fold change > 4), and the results showed that the expression patterns of these genes were consistent with the results of transcriptome sequencing, indicating that the credibility of the sequencing results. Our data indicated that heat stress induced calcium dyshomeostasis, blocked biogenesis, caused ROS accumulation, impaired the antioxidant system and innate defense, and induced apoptosis through the P53 signaling pathway activated by PEG3, decreased growth and development, and enhanced organ damage. These data is very important and helpful to elucidate the molecular mechanism of heat stress and finally to find ways to deal with heat stress damage in sheep.

[Analysis of mental state of allergic rhinitis patients in Chengdu city by symptom check list 90 (SCL-90) scale].

Objective: To analyse the mental state of patients with allergic rhinitis (AR) in Chengdu. Methods: One thousand five hundred and thirty-six AR patients from Sichuan Provincial Integrated Traditional Chinese and Western Medicine Hospital, West China Hospital of Sichuan University, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Sichuan People's Hospital, Sichuan Second Hospital of Traditional Chinese Medicine were selected from July 2013 to January 2018. Eight hundred and twenty-seven patients were screened into study group by inclusion and exclusion standards. The symptom check list 90 (SCL-90) was used to group and score the mental state of these patients according to nine classification criteria: gender, BMI, age, marital status, monthly salary, disease duration, living environment, education level and working environment. Then, the scores were compared within groups. Inter-group comparison was made between the study group and the Chinese norm, and the positive factors for psychological disorders were extracted. Four symptoms in the study group, i.e. nasal itching, sneezing, clear discharge and nasal congestion, were scored on the visual analogue scale (VAS). SPSS 19.0 software was used to carry out statistical analysis. Partial correlation analysis was performed between the positive factors and the symptom scores by multiple regression statistical method. Results: The total score of SCL-90 in the study group was 2.64±0.25, which was accorded with mild to moderate mental health impairment. There were 124 (15.0%) without mental health damage, 176 (21.3%) with mild damage, 474 (57.3%) with mild to moderate damage, 41 (5.0%) with moderate to severe damage and 12 (1.4%) with severe damage. The in-group comparison showed that the top three categories of different items were the living environment, gender and working environment. The scores of somatization, obsessive-compulsive symptoms, interpersonal sensitivity, depression, anxiety, psychosis, other (sleep, diet) and total average score of urban residents were higher than that of country residents (3.29±0.61 vs 2.65±0.50, 2.81±0.77 vs 2.05±0.38, 3.10±0.19 vs 2.49±0.67, 3.40±0.84 vs 2.49±0.70, 3.04±0.64 vs 2.33±0.51, 3.02±0.55 vs 2.40±0.77, 3.40±0.41 vs 2.52±0.77, 2.91±0.11 vs 2.29±0.40, Z value was 4.88, 5.25, 4.57, 5.91, 5.09, 4.63, 5.55, -4.55, respectively, all P<0.05). Women scored higher than man for somatization, interpersonal sensitivity, depression and others (2.66±0.51 vs 2.00±0.45, 3.37±0.47 vs 2.63±0.51, 3.44±0.57 vs 2.85±0.52, 3.47±0.36 vs 2.76±0.45, Z value was -5.10, -5.51, -4.86, -5.28, respectively, all P<0.05). The scores of somatization, interpersonal sensitivity, psychosis and other (sleep, diet) were higher in the indoor group than those in the outdoor group (3.49±0.64 vs 2.78±0.46, 3.33±0.30 vs 2.56±0.68, 3.28±0.60 vs 2.67±0.31, 3.50±0.85 vs 2.85±0.37, Z value was 5.31, 5.79, 4.89, 5.00, respectively, all P<0.05). The outdoor group scored higher on obsessive-compulsive symptoms, anxiety and hostility (3.44±0.40 vs 2.83±0.35, 3.40±0.50 vs 2.57±0.93, 3.34±0.88 vs 2.69±0.56, Z value was 4.96, 6.22, 5.08, respectively, all P<0.05). The inter-group comparison found that depression, anxiety, psychosis and other (sleep, diet) could be partially correlated with VAS scores as 4 positive factors. The results of partial correlation analysis showed that depression was positively correlated with sneezing and nasal runny discharge, anxiety was positively correlated with nasal itching and nasal obstruction, psychosis was positively correlated with nasal itching and sneezing, and other (sleep, diet) was positively correlated with nasal runny discharge and nasal obstruction. Conclusion: AR patients have mild to moderate mental health impairments, which are correlated with AR symptoms.

[Factors related to thyroid carcinoma in Zhejiang province: a matched case-control study].

Objective: To explore the influencing factors related to thyroid carcinoma. Methods: Matched by sex, age and original residential areas, 659 pairs of cases and controls were recruited and studied. Methods including both single factor analysis and multivariate conditional logistic regression analysis were carried out to identify the influencing factors. Results: Multivariate conditional logistic regression analysis showed that higher education, being diabetic, alcohol intake, tea drinking, occupational physical activity and the frequency of eating fishes/eggs etc., were potentially protective to thyroid carcinoma. Depression, personal history of CT examination and less salt intake seemed to be risk factors on thyroid carcinoma. For males, factors as alcohol intake, tea drinking, occupational physical activity and frequent egg-eating appeared protective. For females, higher education, diabetes, tea drinking, occupational physical activity, frequent consumption of fishes/eggs, short duration of menstruation appeared as possibly protective. Conclusion: Higher education, diabetes, alcohol intake, tea drinking, occupational physical activity, frequent consumption of fishes/eggs, depression, personal history of CT examination and less salt intake served as potential influencing factors to thyroid carcinoma.

[Analysis of infection and drug-resistance in 6 107 cases of extrapulmonary tuberculosis in Chengdu area].

Objective: To analyze the infection and drug resistance of extrapulmonary tuberculosis, in order to provide useful references for prevention and treatment of tuberculosis (TB). Methods: A total of 6 107 suspected cases of extrapulmonary tuberculosis, including lymph node, pleural, peritoneal, meningeal, urinary tract, and bone tuberculosis, were collected retrospectively from January 2013 to December 2015 in Public Health Clinical Center of Chengdu. There were 4 033 male and 2 074 female patients, and the average age was 37 years.The epidemiological data, infection and drug resistance were analyzed statistically. Results: There were 21 036 cases of mycobacterial culture from January 2013 to December 2015, of which 7 119 were positive(33.8%). Extrapulmonary tuberculosis was found in 6 107 cases, of which 896 were positive, accounting for 12.6% of the positive cases(896/7 119). Culture-positive cases of extra-pulmonary tuberculosis showed a rising trend from 2013 to 2015 (n=174, 310, 421, respectively), and the annual average increasing rate was over 50%.Cases were distributed at younger ages (21-40 years), with an average age of 34 years.There were more male patients, with a male to female ratio of 2.1∶1. The most common site of extrapulmonary tuberculosis was the bone(62/243, 25.5%), followed by lymph node(283/1 297, 21.8%), pleural(289/1 840, 15.7%), urinary tract(63/452, 13.9%), peritoneum(41/303, 13.5%), meningeal(156/1 915, 8.1%)and other types of tuberculosis(2/57, 3.5%). Drug sensitivity test of 896 extrapulmonary tuberculosis strains showed that the resistance rate (from high to low) was in the order of isoniazid, rifampicin, streptomycin, ofloxacin, ciprofloxacin, amikacin, capreomycin and ethambutol. Multidrug-resistant(MDR)and extremely-drug resistant (XDR) rate was 9.7 %(87/896)and 2.0%(17/896)respectively. Conclusions: Cases of extrapulmonary tuberculosis sent for bacterial culture and the positive rate showed a rising trend by years in Chengdu area.The drug resistance level was higher than that reported both in China and abroad.The prevention and treatment of extrapulmonary tuberculosis should be strengthened in this area.

Distribution of tumor stage and initial treatment modality in patients with primary hepatocellular carcinoma.

OBJECTIVE: This study reviewed the distribution of each tumor stage and each type of initial treatment modality among patients with primary hepatocellular carcinoma (HCC) treated at a tertiary tumor hospital between January 2003 and October 2013. METHODS: Baseline data of patients with primary hepatocellular carcinoma treated between January 2003 and October 2013 were retrospectively collected. Tumor stage was determined according to the Barcelona Clinic Liver Cancer (BCLC) staging system and Hong Kong Clinic Liver Cancer (HKLC) staging system. RESULTS: A total of 6241 patients with primary hepatocellular carcinoma were included in the analysis. In accordance with the BCLC, 28.9% of patients were in stage 0/A, 16.2% in stage B, 53.6% in stage C, and 1.3% in stage D. According to the HKLC stage system, 8.4% patients were in stage I, 1.5% in stage IIa, 29.0% in stage IIb, 10.0% in stage IIIa, 33.6% in stage IIIb, 3.4% in stage IVa, 2.5% in stage IVb, 0.2% in stage Va, and 11.4% in stage Vb. Treatment modalities applied to this patient group were as follows: 33.3% of patients underwent hepatic resection, 36.7% underwent transarterial chemoembolization (TACE), 2.2% underwent radiotherapy, 0.9% underwent local ablated therapy, 8.8% underwent systemic chemotherapy, 4.2% underwent traditional herbal medicine therapy, 0.1% underwent targeted drug therapy, and 13.8% received no treatment. Hepatic resection was the most frequent therapy for patients with BCLC 0/A/B disease, and TACE was the initial therapy for patients with BCLC C disease. In the Hong Kong Clinic Liver Cancer staging system, the main treatments for HKLC I to IIIb disease is hepatic resection and TACE. Systemic chemotherapy was the initial therapy for patients with HKLC IVa/IVb disease. Most HKLC Va/Vb patients received traditional Chinese medicine treatment. CONCLUSION: Prevalence of stage BCLC B and C disease was high among our hepatocellular carcinoma patients. In Hong Kong Clinic Liver Cancer staging system, HKLC I to IIIb disease was high among our HCC patients. Hepatic resection and TACE are initial therapies.

In vivo activity of cefquinome against Riemerella anatipestifer using the pericarditis model in the duck.

Cefquinome is a fourth-generation cephalosporin with broad-spectrum antibacterial activity, including activity against enteric gram-negative bacilli such as Riemerella anatipestifer. The pericarditis model was used to examine the pharmacodynamic characteristics of cefquinome against R. anatipestifer. Serum levels of cefquinome following the administration of different doses were determined by LC-MS/MS. Ducks with ca. 10(6) CFU/mL at the initiation of therapy were treated with cefquinome at doses that ranged from 0.0156 to 2 mg/kg of body weight/day (in 3, 6, 12, or 24 divided doses) for 24 h. The percentage of a 24-h dosing interval that the unbound serum cefquinome concentrations exceeded the MIC (fT > MIC) were the pharmacokinetic (PK)-pharmacodynamic (PD) parameter that best correlated with efficacy (R(2) 86.3% for R. anatipestifer, compared with 58.9% for the area under the concentration-time curve/MIC and 10.6% for peak/MIC). A sigmoid Emax model was used to estimate the magnitudes of the %fT > MIC associated with net bacterial stasis, a 1-log10 CFU reduction from baseline, and a 2-log10 CFU reduction from baseline; the corresponding values were (22.5 ± 1.3) %, (35.2 ± 4.5) %, and (42.4 ± 2.7) %. These data showed that treatment with cefquinome results in marked antibacterial effects in vivo against R. anatipestifer and that the host's immunity may also play a key role in the anti-infective therapy process.

Quantitative meta-analysis of grey matter anomalies in neuropathic pain.

BACKGROUND: Increasing neuroimaging studies have revealed grey matter (GM) anomalies of several brain regions by voxel-based morphometry (VBM) studies in patients with neuropathic pain. The changes have been suggested to be related to central sensitization. Our aim was to investigate concurrence across VBM studies to identify whether different subtypes of neuropathic pain share a common pathophysiological basis revealed by structural abnormalities. METHODS: A systematic search of VBM studies of patients with neuropathic pain and healthy controls published in PubMed and Embase databases from January 2000 to March 2014 was conducted. A quantitative meta-analysis of whole-brain VBM studies in patients with neuropathic pain compared with healthy controls was performed by means of effect-size signed differential mapping. RESULTS: Ten studies comprising 240 patients with neuropathic pain and 263 healthy subjects were systematically included in the present study. Compared to healthy controls, the patients showed consistent decreased GM in bilateral anterior insula and thalamus, right superior frontal gyrus and left postcentral gyrus, and increased GM in right medial frontal gyrus and right posterior insula. The results remained largely unchanged in the following jackknife sensitivity analysis. CONCLUSIONS: This meta-analysis shows strong evidence of brain GM anomalies within the pain matrix in patients with neuropathic pain compared with healthy subjects. Further studies are needed to determine whether the reported changes are specific to neuropathic pain or whether they may be common to other chronic pain.

Effect of glutamine on intestinal barrier function following liver transplantation in rats.

OBJECTIVES: Glutamine is an important fuel for intestinal mucosal epithelial cells, and it promotes intestinal mucosal cell differentiation and proliferation. Most liver transplantation (LT) patients suffer from intestinal barrier dysfunction. Whether enteral glutamine supplementation has beneficial effects on intestinal barrier function following LT is not known. We investigated the effect of glutamine (Gln) supplementation on NF-κB and on the intestinal barrier in rats after an allogenic LT with concomitant immunosuppressive therapy. MATERIALS AND METHODS: Inbred Sprague-Dawley rats (n=40) receiving allogenic LT were randomly divided into Gln and control groups (n=20, each). Gln group rats were administered Gln (0.4 g/kg·day) by gastric infusion for 6 days, while control rats received saline. Ten rats from each group were sampled for basal parameters on the 3rd day, prior to LT. The remaining 10 from each group were sampled after receiving LT. Twenty inbred Sprague-Dawley rats were selected as donors. The 20 recipients underwent orthotopic LT after 3 days of treatment and were given immunosuppressive therapy for 6 days post-operation. They were euthanized for sample collection on the 7th day. NF-κB protein in the intestinal mucosa, portal plasma Gln, endotoxin and TNF-α levels, ileocecal sIgA content, bacterial translocation and mucosal ultrastructure were assessed. RESULTS: On the postoperative day 6, the Gln group had increased plasma Gln and ileocecal sIgA (secretory IgA). Gln group also showed improvement in mucosal microvilli structure and had reduced levels of intestinal mucosal NF-κB, portal endotoxin and TNF-α and decreased bacterial translocation as compared to the control group. CONCLUSIONS: Parenteral supplementation of glutamine ameliorated mucosal injury during allogenic LT, and improved intestinal barrier function. These findings suggest that glutamine supplementation may be an effective therapy to ensure successful recovery from liver transplantation.

Predictors of CNS injury as measured by proton magnetic resonance spectroscopy in the setting of chronic HIV infection and CART.

The reasons for persistent brain dysfunction in chronically HIV-infected persons on stable combined antiretroviral therapies (CART) remain unclear. Host and viral factors along with their interactions were examined in 260 HIV-infected subjects who underwent magnetic resonance spectroscopy (MRS). Metabolite concentrations (NAA/Cr, Cho/Cr, MI/Cr, and Glx/Cr) were measured in the basal ganglia, the frontal white matter, and gray matter, and the best predictive models were selected using a bootstrap-enhanced Akaike information criterion (AIC). Depending on the metabolite and brain region, age, race, HIV RNA concentration, ADC stage, duration of HIV infection, nadir CD4, and/or their interactions were predictive of metabolite concentrations, particularly the basal ganglia NAA/Cr and the mid-frontal NAA/Cr and Glx/Cr, whereas current CD4 and the CPE index rarely or did not predict these changes. These results show for the first time that host and viral factors related to both current and past HIV status contribute to persisting cerebral metabolite abnormalities and provide a framework for further understanding neurological injury in the setting of chronic and stable disease.

Dissolution and pharmacokinetic properties of alkaloids and flavonoids in a Xiexin multiple-unit drug delivery system.

Alkaloids from Rhizoma coptidis and flavonoids from Radix scutellariae were prepared in 2 separate units using a Xiexin multiple-unit drug delivery system (XXMU) to avoid the interactions that occur in the Xiexin decoction (XXD), which reduce oral bioavailability. Similarity factors (f 2 ) were calculated for the release of each component, and 4 equations were fitted to describe the release mechanism. In vivo pharmacokinetic studies were undertaken in rats to compare orally administered XXMU to a XXD powder suspension. No significant differences in the in vitro release properties of the 4 main components were observed between the individual pellets and the entire XXMU delivery system. The release mechanisms for coptisine, berberine, palmatine and baicalin from XXMU involved non-Fickian transport, non-Fickian transport, Fickian diffusion and zero-order release, respectively. The AUC(0-6 h), Cmax1(0-0.5 h) and Cmax3(2-6 h) values for berberine were significantly higher for XXMU than for XXD, but significant differences between the 2 dosage forms were not observed for pharmacokinetic parameters of baicalin. Interactions between flavonoids and alkaloids were reduced in XXMU, and the oral bioavailability of berberine was improved. The successful application of a multiple-unit drug delivery system may provide a method to improve the oral bioavailability of traditional Chinese medicines with multiple components.

Adjuvant therapy options following curative treatment of hepatocellular carcinoma: a systematic review of randomized trials.

AIMS: Numerous postoperative therapies for preventing recurrence of hepatocellular carcinoma (HCC) have been reported, but their efficacy remains controversial and knowledge about adverse effects is limited. A systematic review of randomized controlled trials (RCTs) was performed to gain a comprehensive picture of the efficacy and risks of these therapies. METHODS: MEDLINE, EMBASE and the Cochrane Library were systematically searched through July 2011. Risk ratios (RRs) and 95% confidence intervals (CIs) were calculated. RESULTS: A total of 2989 patients from 28 RCTs involving 10 postoperative therapies were included. For interferon therapy, the estimated RR for the 2-year recurrence rate was 0.84 (95% CI 0.73-0.97, P = 0.02) and the overall survival (OS) was 1.15 (95% CI 1.07-1.22, P < 0.001). Postoperative therapy with the vitamin K2 analog did not lead to a significant reduction in the 1-year recurrence rate, with a pooled RR of 0.60 (95% CI 0.28-1.27, P = 0.18). However, it did slightly improve the 1-year OS, with a pooled RR of 1.03 (95% CI 1.00-1.05, P = 0.03). Transarterial chemotherapy with or without embolization, adoptive immunotherapy and heparanase inhibitor PI-88 therapy may delay tumor recurrence. The effects of acyclic retinoid, lipiodol-iodine-131 and tumor vaccine treatment were promising but require further study. All postoperative therapies except interferon administered intramuscularly were well tolerated by the majority of patients. CONCLUSIONS: Use of adjuvant interferon is definitely associated with an increase in OS. Postoperative therapies involving acyclic retinoid, lipidol-iodine-131, or tumor vaccine may improve the OS of patients with HCC after curative treatment.

Inhibition of tumor cell proliferation and induction of apoptosis in human lung carcinoma 95-D cells by a new sesquiterpene from hairy root cultures of Artemisia annua.

Artemisia annua is a rich source of many bioactive substances, and in our recent work, a new sesquiterpene, (Z)-7-acetoxy-methyl-11-methyl-3-methylene-dodeca-1,6,10-triene (AMDT), was isolated and identified from hairy roots culture of A. annua, and its bioactivity was characterized in this work. AMDT showed moderate cytotoxic activities against the human tumor cell lines of HO8910 (ovary), 95-D (lung), QGY (liver) and HeLa (cervix) by MTT assay, whose IC(50) values were ranged within 52.44-73.3 microM. As lung cancer is the No. 1 killer of global cancer patients, our interest is to investigate the ability of AMDT in inducing apoptosis of 95-D tumor cells. The 95-D cell growth was inhibited by AMDT, and the flow cytometry analysis showed its cell cycle was arrested in the G1 phase. The apoptotic rate of the cells increased in a dose-dependent manner. AMDT lowered the mitochondrial membrane potential and increased the expression of caspase-9 and -3. These results revealed that AMDT could efficiently induce 95-D cell apoptosis through mitochondrial dependent pathway, and it may be a potential chemotherapeutic agent.

A protein interaction map of Drosophila melanogaster.

Drosophila melanogaster is a proven model system for many aspects of human biology. Here we present a two-hybrid-based protein-interaction map of the fly proteome. A total of 10,623 predicted transcripts were isolated and screened against standard and normalized complementary DNA libraries to produce a draft map of 7048 proteins and 20,405 interactions. A computational method of rating two-hybrid interaction confidence was developed to refine this draft map to a higher confidence map of 4679 proteins and 4780 interactions. Statistical modeling of the network showed two levels of organization: a short-range organization, presumably corresponding to multiprotein complexes, and a more global organization, presumably corresponding to intercomplex connections. The network recapitulated known pathways, extended pathways, and uncovered previously unknown pathway components. This map serves as a starting point for a systems biology modeling of multicellular organisms, including humans.

Effects of a high molecular mass Convolvulus arvensis extract on tumor growth and angiogenesis

BACKGROUND: Plant materials represent promising sources of anti-cancer agents. We developed and tested a novel extract from the ubiquitous plant Convolvulus arvensis. MATERIALS AND METHODS: Convolvulus arvensis components were extracted in boiling water, and small molecules were removed by high-pressure filtration. The extract's biological activity was assessed by measuring its effects on S-180 fibrosarcoma growth in Kun Ming mice and on heparin-induced angiogenesis in chick embryos. We also examined the extract's effects on lymphocytes ex vivo and tumor cell growth in vitro. RESULTS: The extract (primarily proteins and polysaccharides) inhibited tumor growth in a dose-dependent fashion when administered orally. At the highest dose tested, 200 mg/kg/day, tumor growth was inhibited by roughly seventy percent. Subcutaneous or intraperitoneal administration at 50 mg/kg/day also inhibited tumor growth by over seventy percent. The extract's acute LD50 in Kun Ming mice was 500 mg/kg/day when injected, indicating that tumor growth inhibition occurred at non-toxic doses. It inhibited angiogenesis in chick embryos, improved lymphocyte survival ex vivo, and enhanced yeast phagocytosis, but did not kill tumor cells in culture. CONCLUSION: High molecular mass extract deserves further study as an anti-cancer agent.

Biochemical engineering of the production of plant-specific secondary metabolites by cell suspension cultures.

Plant cell culture has recently received much attention as a useful technology for the production of valuable plant-derived secondary metabolites such as paclitaxel and ginseng saponin. The numerous problems that yet bewilder the optimization and scale-up of this process have not been over emphasized. In spite of the great progress recorded in recent years towards the selection, design and optimization of bioreactor hardware, manipulation of environmental factors such as medium components, light irradiation, shear stress and O2 supply needs detailed investigations for each case. Recent advances in plant cell processes, including high-density suspension cultivation, continuous culture, process monitoring, modeling and control and scale-up, are also reviewed in this chapter. Further developments in bioreactor cultivation processes and in metabolic engineering of plant cells for metabolite production are expected in the near future.

Improvement of Panax notoginseng cell culture for production of ginseng saponin and polysaccharide by high density cultivation in pneumatically agitated bioreactors.

A Panax notoginseng cell culture was successfully scaled up from shake flask to 1.0-L bubble column reactor and concentric-tube airlift reactor. High-density bioreactor batch cultivation was carried out using a modified MS medium. The maximum cell density in batch cultures reached 20.1, 21.0 and 24.1 g/L in the shake flask, bubble column and airlift reactors, respectively, and their corresponding biomass productivity was 950, 1140 and 1350 mg/(L x d) for each. The productivity of ginseng saponin was 70, 96 and 99 mg/(L x d) in the flask, bubble column and airlift reactors, respectively; and the polysaccharide productivity reached 104, 119 and 151 mg/(L x d) for each. Furthermore, a fed-batch cultivation strategy was developed on the basis of specific oxygen uptake rate (SOUR), i.e., sucrose feeding before a sharp decrease of SOUR, and the highest cell density of 29.7 g/L was successfully achieved in the airlift bioreactor on day 17 with a very high biomass productivity of 1520 mg/(L x d). The concentrations of ginseng saponin and polysaccharide reached about 2.1 and 3.0 g/L, respectively, and their productivity was 106 (saponin) and 158 mg/(L x d) (polysaccharide). This work successfully demonstrated the high-density bioreactor cultivation of P. notoginseng cells in pneumatically agitated bioreactors and the reproduction of the shake flask culture results in bioreactors. The cell density, biomass productivity, production titer and productivity of both ginseng saponin and polysaccharide obtained here were the highest that have been reported on a reactor scale for all the ginseng species.

Green tea polyphenol extract attenuates inflammation in interleukin-2-deficient mice, a model of autoimmunity.

Green tea polyphenols (GrTP) have been previously shown to decrease endotoxin-induced tumor necrosis factor-alpha production and lethality in mice. Our present studies demonstrate that GrTP inhibit inflammatory responses and may be useful in treating chronic inflammatory states, such as inflammatory bowel disease. In this preliminary study, we examined whether GrTP decrease disease activity in interleukin-2-deficient (IL-2(-/-) mice. Eight-week old IL-2(-/-) C57BL/6J mice who were fed nonpurified diet were randomly assigned to receive water with GrTP (5 g/L) or water alone (control) for up to 6 wk. After 1 wk, explant colon and lamina propria lymphocyte (LPL) cultures were established from a subgroup of mice and supernatants collected. Culture supernatants from GrTP-treated mice showed decreased spontaneous interferon-gamma and tumor necrosis factor-alpha secretion compared with that of controls. At 6 wk, the GrTP group had less severe colitis as demonstrated by lower histologic scores and wet colon weights. This was associated with lower plasma levels of serum amyloid A, increased weight gain and improved hematocrits. These results show that GrTP attenuated inflammation in IL-2(-/-) mice and suggest a role for GrTP in treating chronic inflammatory diseases such as inflammatory bowel disease.

fMRI of auditory stimulation with intermolecular double-quantum coherences (iDQCs) at 1.5T.

An intermolecular double-quantum coherence (iDQC) imaging technique was used to study auditory activation in the human brain at 1.5T with a dual temporal lobe surface phased array coil and a quadrature head coil. Preliminary results demonstrate that it is feasible to obtain auditory activation maps using iDQC imaging at 1.5T, both in individual subjects using the surface coil array and with multisubject averaging of data using the head coil. The most robust activation map was obtained when a spin-echo (SE) acquisition was combined with an iDQC excitation. Since SE with conventional single quantum coherence (SQC) and similar parameters showed much reduced activation in spite of its higher signal-to-noise ratio (SNR), it was determined that activation resulting from the SE-iDQC acquisition almost entirely originates from iDQCs. In addition, the fact that the robust activation was obtained using signals at an evolution time more sensitive to changes in magnetic susceptibilities also suggests the sensitivity of iDQCs to the BOLD effect upon activation. iDQCs provide a novel MRI method which is potentially more sensitive to the BOLD effect traditionally measured with SQC. Magn Reson Med 45:356-364, 2001.

Effects of amelogenin on the transforming surface microstructures of Bioglass in a calcifying solution.

Topographies of a bioactive glass (45S5 type Bioglass(R)) during 0-4 h of immersion in a supersaturated calcifying solution (SCS) and the SCS containing recombinant porcine amelogenin rP172 (SCS(rP172)) were observed by atomic force microscopy. Other techniques including X-ray diffraction, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, and transmission electron microscopy were used for some complementary microstructural investigations. The smooth Bioglass surface changed to be very rough after 0.5 h of SCS immersion because of glass network dissolution. Spherical silica-gel particles with diameters of 150-300 nm consisting of substructures of 20-60 nm across had formed on the sample surfaces after 1 h of SCS immersion. The chemisorption of amorphous calcium phosphate and crystallization of nanophase apatite were seen to occur epitaxially on the silica-gel structures during 1-4 h of SCS immersion. During the first 0.5 h of SCS(rP172) immersion, more than 95% of rP172 protein in solution was adsorbed onto the sample surfaces and generated spherical assemblies of 10-60 nm diameters. During 0.5-4 h of SCS(rP172) immersion, the protein assemblies of rP172 remarkably induced the formation of orientated silica-gel plates (approximately 100-nm wide and 50-nm thick) and subsequently of long and thin apatite needle crystals. The recombinant amelogenin rP172-modulated apatite crystals resembled those formed in the early stage of tooth enamel biomineralization, suggesting the functional roles of amelogenins during the oriented growth of enamel crystallites and a great potential for amelogenins in applications designed to fabricate enamel-like calcium phosphate biomaterials.