Microbes as a production host to produce natural activecompounds from mushrooms: biosynthetic pathway elucidationand metabolic engineering.

Mushrooms are abundant in bioactive natural compounds. Due to strict growth conditions and long fermentation-time, microbe as a production host is an alternative and sustainable approach for the production of natural compounds. This review focuses on the biosynthetic pathways of mushroom originated natural compounds and microbes as the production host for the production of the above natural compounds.

Benzene exposure-response and risk of lymphoid neoplasms in Chinese workers: A multicenter case-cohort study.

BACKGROUND: While international agreement supports a causal relationship of benzene exposure with acute myeloid leukemia, there is debate about benzene and lymphoid neoplasm risks. METHODS: In a case-cohort study with follow-up of 110 631 Chinese workers during 1972-1999, we evaluated benzene exposure-response for non-Hodgkin lymphoma (NHL), lymphoid leukemias (LL), acute lymphocytic leukemia (ALL), and total lymphoid neoplasms (LN). We estimated benzene exposures using state-of-the-art hierarchical modeling of occupational factors calibrated with historical routine measurements and evaluated cumulative exposure-response using Cox regression. RESULTS: NHL and other specified LN were increased in exposed vs unexposed workers. However, there was no evidence of exposure-response for NHL or other specified LN. Based on a linear exposure-response, relative risks at 100 parts per million-years (RR at 100 ppm-years) for cumulative benzene exposure using a 2-year lag (exposure at least 2 years before the time at risk) were 1.05 for NHL (95 percent confidence interval (CI) = 0.97, 1.27; 32 cases), 1.11 for LL (95% CI < 0, 1.66; 12 cases), 1.21 for ALL (95% CI < 0, 3.53; 10 cases), and 1.02 for total LN (95% CI < 0, 1.16; 49 cases). No statistically significant exposure-response trends were apparent for these LN for 2 to <10-year or ≥10-year lags. NHL risks were not significantly modified by sex, age, or year at first exposure, attained age, or time since exposure. CONCLUSION: Given the study strengths and limitations, we found little evidence of exposure-response for benzene and NHL, LL, ALL, or total LN, although NHL and other specified LN were increased in exposed vs unexposed individuals.

Benzene Exposure Response and Risk of Myeloid Neoplasms in Chinese Workers: A Multicenter Case-Cohort Study.

BACKGROUND: There is international consensus that benzene exposure is causally related to acute myeloid leukemia (AML), and more recent evidence of association with myelodysplastic syndromes (MDS). However, there are uncertainties about the exposure response, particularly risks by time since exposure and age at exposure. METHODS: In a case-cohort study in 110 631 Chinese workers followed up during 1972-1999 we evaluated combined MDS/AML (n = 44) and chronic myeloid leukemia (n = 18). We estimated benzene exposures using hierarchical modeling of occupational factors calibrated with historical routine measurements, and evaluated exposure response for cumulative exposure and average intensity using Cox regression; P values were two-sided. RESULTS: Increased MDS/AML risk with increasing cumulative exposure in our a priori defined time window (2 to <10 years) before the time at risk was suggested (Ptrend = 08). For first exposure (within the 2 to <10-year window) before age 30 years, the exposure response was stronger (P = .004) with rate ratios of 1.12 (95% confidence interval [CI] = 0.27 to 4.29), 5.58 (95% CI = 1.65 to 19.68), and 4.50 (95% CI = 1.22 to 16.68) for cumulative exposures of more than 0 to less than 40, 40 to less than 100, and at least 100 ppm-years, respectively, compared with no exposure. There was little evidence of exposure response after at least 10 years (Ptrend = .94), regardless of age at first exposure. Average intensity results were generally similar. The risk for chronic myeloid leukemia was increased in exposed vs unexposed workers, but appeared to increase and then decrease with increasing exposure. CONCLUSION: For myeloid neoplasms, the strongest effects were apparent for MDS/AML arising within 10 years of benzene exposure and for first exposure in the 2 to less than 10-year window before age 30 years.

Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek's disease virus.

BACKGROUND: Chicken erythrocytes are involved in immunity through binding of toll-like receptors (TLRs) with their ligands to activate downstream signaling and lead to cytokine production in erythrocytes. Some avian ß-defensins (AvBDs) are constitutively expressed in tissues and some others can be induced by various bacteria and viruses. However, the expression of AvBDs in erythrocytes has not yet been studied extensively. RESULTS: The transcripts of eight AvBDs (AvBD1 to AvBD7, and AvBD9) and liver-expressed antimicrobial peptide-2 (LEAP-2) were found in normal chicken erythrocytes. The expression levels of AvBD2, 4 and 7 were significantly increased (P < 0.01), whereas the levels of AvBD1, 6 and 9 were significantly decreased (P < 0.01) after Marek's disease virus (MDV) infection. The mRNA expression level of LEAP-2 was not significantly changed after MDV infection. Highest viral nucleic acid (VNA) of MDV in the feather tips among the tested time points was found at 14 days post-infection (d.p.i.). In addition, 35 MD5-related gene segments were detected in the erythrocytes at 14 d.p.i. by transcriptome sequencing. CONCLUSIONS: These results suggest that the AvBDs in chicken erythrocytes may participate in MDV-induced host immune responses.

Adductomic signatures of benzene exposure provide insights into cancer induction.

Although benzene has long been recognized as a cause of human leukemia, the mechanism by which this simple molecule causes cancer has been problematic. A complicating factor is benzene metabolism, which produces many reactive intermediates, some specific to benzene and others derived from redox processes. Using archived serum from 20 nonsmoking Chinese workers, 10 with and 10 without occupational exposure to benzene (exposed: 3.2-88.9 ppm, controls: 0.002-0.020 ppm), we employed an adductomic pipeline to characterize protein modifications at Cys34 of human serum albumin, a nucleophilic hotspot in extracellular fluids. Of the 47 measured human serum albumin modifications, 39 were present at higher concentrations in benzene-exposed workers than in controls and many of the exposed-control differences were statistically significant. Correlation analysis identified three prominent clusters of adducts, namely putative modifications by benzene oxide and a benzene diolepoxide that grouped with other measures of benzene exposure, adducts of reactive oxygen and carbonyl species, and Cys34 disulfides of small thiols that are formed following oxidation of Cys34. Benzene diolepoxides are potent mutagens and carcinogens that have received little attention as potential causes of human leukemia. Reactive oxygen and carbonyl species-generated by redox processes involving polyphenolic benzene metabolites and by Cyp2E1 regulation following benzene exposure-can modify DNA and proteins in ways that contribute to cancer. The fact that these diverse human serum albumin modifications differed between benzene-exposed and control workers suggests that benzene can increase leukemia risks via multiple pathways involving a constellation of reactive molecules.

Current Status of Standardization of Traditional Chinese Medicine in China.

As an important component of traditional medicine, Traditional Chinese Medicine (TCM) is widely spread and applied in more than 100 countries across the world. The standardization of TCM is very important for the international application of Chinese medicine. In this paper, we have explained and analyzed the standardization situations of TCM in China with the purpose of providing reference for standardization and international development of TCM.

Retrospective benzene exposure assessment for a multi-center case-cohort study of benzene-exposed workers in China.

Quality of exposure assessment has been shown to be related to the ability to detect risk of lymphohematopoietic disorders in epidemiological investigations of benzene, especially at low levels of exposure. We set out to build a statistical model for reconstructing exposure levels for 2898 subjects from 501 factories that were part of a nested case-cohort study within the NCI-CAPM cohort of more than 110,000 workers. We used a hierarchical model to allow for clustering of measurements by factory, workshop, job, and date. To calibrate the model we used historical routine monitoring data. Measurements below the limit of detection were accommodated by constructing a censored data likelihood. Potential non-linear and industry-specific time-trends and predictor effects were incorporated using regression splines and random effects. A partial validation of predicted exposures in 2004/2005 was performed through comparison with full-shift measurements from an exposure survey in facilities that were still open. Median cumulative exposure to benzene at age 50 for subjects that ever held an exposed job (n=1175) was 509 mg/m(3) years. Direct comparison of model estimates with measured full-shift personal exposure in the 2004/2005 survey showed moderate correlation and a potential downward bias at low (<1 mg/m(3)) exposure estimates. The modeling framework enabled us to deal with the data complexities generally found in studies using historical exposure data in a comprehensive way and we therefore expect to be able to investigate effects at relatively low exposure levels.

A retrospective cohort study of cause-specific mortality and incidence of hematopoietic malignancies in Chinese benzene-exposed workers.

Benzene exposure has been causally linked with acute myeloid leukemia (AML), but inconsistently associated with other hematopoietic, lymphoproliferative and related disorders (HLD) or solid tumors in humans. Many neoplasms have been described in experimental animals exposed to benzene. We used Poisson regression to estimate adjusted relative risks (RR) and the likelihood ratio statistic to derive confidence intervals for cause-specific mortality and HLD incidence in 73,789 benzene-exposed compared with 34,504 unexposed workers in a retrospective cohort study in 12 cities in China. Follow-up and outcome assessment was based on factory, medical and other records. Benzene-exposed workers experienced increased risks for all-cause mortality (RR = 1.1, 95% CI = 1.1, 1.2) due to excesses of all neoplasms (RR = 1.3, 95% CI = 1.2, 1.4), respiratory diseases (RR = 1.7, 95% CI = 1.2, 2.3) and diseases of blood forming organs (RR = ∞, 95% CI = 3.4, ∞). Lung cancer mortality was significantly elevated (RR = 1.5, 95% CI = 1.2, 1.9) with similar RRs for males and females, based on three-fold more cases than in our previous follow-up. Significantly elevated incidence of all myeloid disorders reflected excesses of myelodysplastic syndrome/acute myeloid leukemia (RR = 2.7, 95% CI = 1.2, 6.6) and chronic myeloid leukemia (RR = 2.5, 95% CI = 0.8, 11), and increases of all lymphoid disorders included excesses of non-Hodgkin lymphoma (RR = 3.9, 95%CI = 1.5, 13) and all lymphoid leukemia (RR = 5.4, 95%CI = 1.0, 99). The 28-year follow-up of Chinese benzene-exposed workers demonstrated increased risks of a broad range of myeloid and lymphoid neoplasms, lung cancer, and respiratory diseases and suggested possible associations with other malignant and non-malignant disorders.

Development and validation of an attenuated Mycoplasma hyopneumoniae aerosol vaccine.

Mycoplasma hyopneumoniae (M. hyopneumoniae) causes a chronic respiratory disease with high morbidity and low mortality in swine, and has been presented as a major cause of growth retardation in the swine industry. Aerosol vaccination presents a needle free, high throughput, and efficient platform for vaccine delivery, and has been widely applied in poultry vaccination. However, aerosol vaccines have rarely been used in swine vaccination primarily because the long and curving respiratory track of swine presents a barrier for vaccine particle delivery. To develop an effective M. hyopneumoniae aerosol vaccine, three major barriers need to be overcome: to optimize particle size for aerosol delivery, to maintain the viability of mycoplasma cells in the vaccine, and to optimize the environmental conditions for vaccine delivery. In this study, an aerosol mycoplasma vaccine was successfully developed based on a conventional live attenuated M. hyopneumoniae vaccine. Specifically, the Pari LCD nebulizer was used to produce an aerosol vaccine particle size less than 5 µm; and a buffer with 5% glycerol was developed and optimized to prevent inactivation of M. hyopneumoniae caused by aerosolization and evaporation. Before nebulization, the room temperature and relative humidity were control to 20-25 °C and 70-75%, respectively, which helped maintain the viability of aerosol vaccine. Animal experiments demonstrated that this newly developed aerosol vaccine was effectively delivered to swine low respiratory track, being confirmed by nested-PCR, in situ hybridization and scanning electron microscope. Moreover, M. hyopneumoniae specific sIgA secretion was detected in the nasal swab samples at 14 days post-immunization. To our knowledge, this is the first report on a live M. hyopneumoniae aerosol vaccine.

[Discussion on necessity of standardization for nomenclature and location of extraordinary acupoints].

With retrieval of articles on extraordinary acupoint that were published in domesticin recent five years, one hundred and eight articles of clinical application are screened out and one hundred and twenty-three extraordinary acupoints that are extensively recognized are collected. Of those acupoints, 23 acupoints are included in the latest national standard. Of the rest 100 extraordinary acupoints, 48 acupoints are located on the running courses of fourteen meridians, 4 acupoints are shared with the meridian points and the other 52 acupoints have not been clarified to be located on the running courses of meridians based on the literature data. It is found in the collection of these acupoints that there are many extraordinary acupoints that are extensively used in clinical practice. But the nomenclatures and locations of acupoints have not been unified. Hence, a further standardization on these aspects is anticipated.

[Current situation of the standardization of acupuncture and moxibustion in Taiwan].

The current situation of the standardization of acupuncture and moxibustion in the Taiwan region is introduced in this paper from the three aspects, named the development state of standard of acupuncture and moxibustion in Taiwan, the implementation of Taiwan district standard and the standardization of acupuncture and moxibustion in Taiwan. At present, the relevant standards of acupuncture and moxibustion in Taiwan just include the standard operation procedure of acupuncture and moxibustion, the reference guideline of the safe operation in the medical service centers of traditional Chinese medicine, and the faculty standard of Chinese medicine hospital, etc. It is concluded that the current situation of the standardization of acupuncture and moxibusiton presented the weak awareness of the standardization of acupuncture and moxibustion in the industry, insufficient enterprise standard, less-quantity of the implemented standards and narrow coverage.

Crystallization and preliminary X-ray analysis of argininosuccinate lyase from Streptococcus mutans.

Argininosuccinate lyase (ASL) is an important enzyme in arginine synthesis and the urea cycle, which are highly conserved from bacteria to eukaryotes. The gene encoding Streptococcus mutans ASL (smASL) was amplified and cloned into expression vector pET28a. The recombinant smASL protein was expressed in a soluble form in Escherichia coli strain BL21 (DE3) and purified to homogeneity by two-step column chromatography. Crystals suitable for X-ray analysis were obtained and X-ray diffraction data were collected to a resolution of 2.5 Å. The crystals belonged to space group R3, with unit-cell parameters a = b = 254.5, c = 78.3 Å.

[Study on Chinese herb Astragalus membranceus by FTIR fingerprint].

UNLABELLED: The aim of the study is to find new approach to identifying different growth years and different producing areas of astragalus membranceus simply and rapidly. The FTIR spectrometry, which is accurate, simple and efficient and has high resolution, was used to obtain the sample's FTIR spectrometry. After simple separation and extraction, the drying powder of the sample was detected. In 22 samples, there were 9 common peaks for identifying the authenticity of astragalus membranceus and 5 different peaks. According to the number, shape, and relative intensity of the peak, different growth years and different producing areas of astragalus membranceus can be distinguished. CONCLUSION: It is convenient and fast to identify different growth years and different producing areas of astragalus membranceus by FTIR fingerprint.

[Debating some issues on the national standard Standardized Manipulation of Acupuncture and Moxibustion, part 3: Auricular Acupuncture].

The main difficulties and disputable problems on develping the national standard Standardized Manipulation of Acupuncture and Moribustion, Part 3: Auricular Acupuncture are debated in this paper from the three aspects of its meaning, scientific basis and key point. The difficult points mainly include manipulation standandization of auricular acupuncture, basis and science of the standards stipulated, selection of technical terms and definition, and why only the four kinds of manipulations are used for auricular acupuncture. Finally, the authors suggest that the Standardized Manipulation of Acupuncture and Moxibustion, Part 3: Auricular Acupuncture needs further to be studied and improved.

Crystallization and preliminary X-ray analysis of three dUTPases from Gram-positive bacteria.

All organisms examined to date possess a dUTPase that performs the important function of efficiently hydrolyzing dUTP to dUMP in order to prevent the incorporation of dUTP into DNA. Three putative dUTPases from Gram-positive bacteria have been studied in this work. Two dUTPase-encoding genes, yncF and yosS, have been identified in Bacillus subtilis. The gene dut, encoding dUTPase from the dental pathogen Streptococcus mutans, was amplified from S. mutans genomic DNA. The three genes were cloned into expression vectors and overexpressed at high levels in Escherichia coli. Each protein was purified in two steps using chromatographic methods. Crystals of the YosS and YncF proteins and of S. mutans dUTPase were obtained using the vapour-diffusion method. X-ray diffraction data sets were collected from crystals of selenomethionine-labelled YosS and S. mutans dUTPase to resolutions of 2.3 and 1.7 A, respectively. The crystal of native YncF diffracted to 2.7 A resolution.

[Approach to some problems in the national standard Standardized Manipulation of Acupuncture and Moxibustion, Part IV, Three-Edged Needle].

Main difficulties and problems in working out the national standard Manipulation Standardization of Acupuncture and Moribustion Techniques for Three-edged Needle are discussed. The difficult points include selection of technical terms; standardization and characteristics of Chinese medicine; quantification of bleeding amount in three-edged needle therapy; basis and science of the standards stipulated. The problems that need further be studied and perfected for Manipulation Standardization of Acupuncture and Moxibustion Techniques for Three-edged Needle are put forward in the present paper.

Open-closed conformational change revealed by the crystal structures of 3-keto-L-gulonate 6-phosphate decarboxylase from Streptococcus mutans.

The 3-keto-L-gulonate 6-phosphate decarboxylase (KGPDC) catalyses the decarboxylation of 3-keto-L-gulonate 6-phosphate to L-xylulose in the presence of magnesium ions. The enzyme is involved in L-ascorbate metabolism and plays an essential role in the pathway of glucuronate interconversion. Crystal structures of Streptococcus mutans KGPDC were determined in the absence and presence of the product analog D-ribulose 5-phosphate. We have observed an 8 A alphaB-helix movement and other structural rearrangements around the active site between the apo-structures and product analog bound structure. These drastic conformational changes upon ligand binding are the first observation of this kind for the KGPDC family. The flexibilities of both the alpha-helix lid and the side chains of Arg144 and Arg197 are associated with substrate binding and product releasing. The open-closed conformational changes of the active site, through the movements of the alpha-helix lid and the arginine residues are important for substrate binding and catalysis.

[A brief talk on characteristics of Japanese collateral puncture].

The characteristics of Japanese collateral puncture are expounded from history evolution and technical feature. The position of pricking blood and blood-letting volume for the three Japanese collateral puncture, Japanese collateral puncture at Jing-well points, at fine collaterals and at the skin are emphatically discussed and briefly compared with Chinese pricking blood therapy, so as to better develop the collateral puncture therapy.

[Summarization and analysis on acupuncture-related articles embodied in Medline data-base in 2006].

OBJECTIVE: To understand the situation of studies and development of international acupuncture and moxibustion in 2006 by analysis on the acupuncture-related articles embodied in Medline data-base in 2006. METHODS: A total of 881 articles in Medline data-base of the ISI knowledge Web were retrieved by "acupuncture", "moxibustion", "acupressure" and "cupping" as key words, then 761 articles excluding unrelated 120 ones were analyzed and concluded for distribution of the article sources, periodicals source, type, research contents and methods. RESULTS: Most of the articles are from China, Britain and America. The journal of the highest impact factor of 25.8 is Lancet. Acupuncture and moxibustion can treat various diseases, but primarily for relieving pain. CONCLUSION: Acupuncture and moxibustion are still a part of CAM for clinical practice abroad. Most of the experiments confirm the effects of acupuncture and moxibustion, but there are some different views on some diseases. And various countries make a lot work about mechanism research of acupuncture effect, showing the feature of large range.

[Synthesis and bioactivies of salicylic acid-g-chitosan derivatives].

To prepare the derivatives of salicylic acid-g-chitosan and study their synergistic and complementary actions, the synergism of anti-inflammatory action of the derivatives was investigated with the experiments of xylene-induces mice ear edema, the analgesic activities by the tartaric emetic-induced mice twist test and the hot-plate test, and the complementary effects between salicylic acid and chitosan through morphological changes of stomach mucous membrane of rat, separately. The anti-inflammatory activities of salicylic acid-g-chitosan derivatives for anti-inflammatory activities were more potent than that of salicylic acid and chitosan and dexamethasone cream in external use, and more potent than that of aspirin orally. However, immediate analgesic activity of the derivatives was lower than that of aspirin and persistent activity was similar as that of aspirin. And the stomach mucous membrane morphology change of the derivatives was much milder than that of aspirin. The salicylic acid grafted chitosan derivatives showed synergistic and complementary effect on the anti-inflammatory and analgesic activities and so on.