Lignin bioconversion based on genome mining for ligninolytic genes in Erwinia billingiae QL-Z3.

BACKGROUND: Bioconversion of plant biomass into biofuels and bio-products produces large amounts of lignin. The aromatic biopolymers need to be degraded before being converted into value-added bio-products. Microbes can be environment-friendly and efficiently degrade lignin. Compared to fungi, bacteria have some advantages in lignin degradation, including broad tolerance to pH, temperature, and oxygen and the toolkit for genetic manipulation. RESULTS: Our previous study isolated a novel ligninolytic bacterial strain Erwinia billingiae QL-Z3. Under optimized conditions, its rate of lignin degradation was 25.24% at 1.5 g/L lignin as the sole carbon source. Whole genome sequencing revealed 4556 genes in the genome of QL-Z3. Among 4428 protein-coding genes are 139 CAZyme genes, including 54 glycoside hydrolase (GH) and 16 auxiliary activity (AA) genes. In addition, 74 genes encoding extracellular enzymes are potentially involved in lignin degradation. Real-time PCR quantification demonstrated that the expression of potential ligninolytic genes were significantly induced by lignin. 8 knock-out mutants and complementary strains were constructed. Disruption of the gene for ELAC_205 (laccase) as well as EDYP_48 (Dyp-type peroxidase), ESOD_1236 (superoxide dismutase), EDIO_858 (dioxygenase), EMON_3330 (monooxygenase), or EMCAT_3587 (manganese catalase) significantly reduced the lignin-degrading activity of QL-Z3 by 47-69%. Heterologously expressed and purified enzymes further confirmed their role in lignin degradation. Fourier transform infrared spectroscopy (FTIR) results indicated that the lignin structure was damaged, the benzene ring structure and groups of macromolecules were opened, and the chemical bond was broken under the action of six enzymes encoded by genes. The abundant enzymatic metabolic products by EDYP_48, ELAC_205 and ESOD_1236 were systematically analyzed via liquid chromatography-mass spectrometry (LC-MS) analysis, and then provide a speculative pathway for lignin biodegradation. Finally, The activities of ligninolytic enzymes from fermentation supernatant, namely, LiP, MnP and Lac were 367.50 U/L, 839.50 U/L, and 219.00 U/L by orthogonal optimization. CONCLUSIONS: Our findings provide that QL-Z3 and its enzymes have the potential for industrial application and hold great promise for the bioconversion of lignin into bioproducts in lignin valorization.

[The campaign of returning to the ancient in sphygmology in the Republican Period as viewed from Mai xue cong shu (Series of Pulse Taking)].

From the Southern and Northern Dynasties to the beginning of the Northern Song Dynasty, the models of "master and apprentice" and "physician of long family tradition for generations" were the main ways for teaching medical knowledge. With the rapid amassment of medical books in the Due to the rapid transmission of western science and technology into China in the Republican period, the art of pulse taking had been treated as unscientific. Yao Xinyuan advocated the recovery of ancient pulse taking of the three-portion approach, i.e., taking the pulses at the neck, hands, and feet for comparison. To spread this idea, Yao and Zhang Ziying compiled the Mai xue cong shu (Series of Pulse Taking) in 1937-1947. Altogether 4 issues were published carrying 29 articles concerning the theoretical exploration, experimental research, and clinical practice. Their thinking and approaches were influential to the study and development of modern sphygmology.

[The first national Whole-time Training Class for Western Physicians Learning Traditional Chinese Medicine in China].

On 19 October, 1955, at the time when the Academy of Traditional Chinese Medicine affiliated with the Ministry of Health was established, the opening ceremony of the first Whole-time Training Class for Western Physicians Learning Systematically Traditional Chinese Medicine was also held. Altogether 84 new graduates from western medical colleges and skilled doctors of western medicine from all over the country were registered. During the 2.5 year term of training, the students learnt 16 professional courses and then practiced at the clinic, and graduated with gratifying results in July, 1958. On 11 October the same year, Chairman Mao made an important instruction to the report submitted by the Leading Party Group of the Ministry of Health, "A Report to the Party Central Committee about the situation, results and experience of Whole-time Training Class for Western Physicians Learning Traditional Chinese Medicine", by pointing out that "in the future, the Training Class for Western Physicians Learning Traditional Chinese Medicine should be held across the country hereafter. This is a top issue and is not to be ignored".

Role of Rheum Polysaccharide in the cytokines produced by peripheral blood monocytes in TLR4 mediated HLA-B27 associated AAU.

PURPOSE: To evaluate the effect of a traditional Chinese medicine, Rheum Polysaccharide (RP), on the in vitro production of tumor necrosis factor alpha (TNF- α ) and interleukin-10 (IL-10) by lipopolysaccharide- (LPS-)stimulated human monocytes from HLA-B27 associated acute anterior uveitis patients of convalescence stage. METHOD: PBMC samples were isolated from 10 HLA-B27 associated acute anterior uveitis, incubated, respectively, and divided into 4 groups as follows: (1) controls, PBS was added in final concentration of 1 mg·L⁻¹, (2) stimulated by LPS, LPS was added in final concentration of 1 mg·L⁻¹, (3) stimulated by LPS + HTA125, 30 minutes before the adding of LPS in final concentration of 1 mg·L⁻¹, the final concentration of 5 mg·L⁻¹ of the HTA125 was added, and (4) stimulated by LPS + RP, 30 minutes before the adding of LPS in final concentration 1 mg·L⁻¹, the final concentration 100 mg·L⁻¹ of the RP was added. Supernatants were used to quantify the amounts of TNF- α and IL-10 released in time course using enzyme-linked immunosorbent assay (ELISA). RESULT: After stimulated by lps, the concentrations of TNF- α and IL-10 in culture supernatants of patients are significantly higher than control group at all time points (P < 0.01). Blockage of TLR-4 by HTA125 can decrease the production of TNF- α and IL-10 compared with lps group (P < 0.01, except at 4 h group of IL-10). Concentration of TNF- α and IL-10 also decreases in the LPS + RP group (P < 0.01) but not so significantly as in the LPS + HTA125 group. CONCLUSION: As anti-TLR4 monoclonal antibodies, rheum Polysaccharide can also inhibit the secretion of cytokines produced by monocytes from HLA-B27 positive AAU patients of convalescence stage.

Preparation of an immunoaffinity column with amino-silica gel microparticles and its application in sample cleanup for aflatoxin detection in agri-products.

This study established an immunoaffinity column for selective extraction of aflatoxins in agri-products. Specifically, the immunoaffinity column was developed by covalently coupling monoclonal antibody 1C11 against aflatoxins to amino-silica gel microparticles and then packing these into a cartridge. The extraction conditions were thoroughly optimized in terms of loading, washing and eluting solutions. Under the optimal conditions, the immunoaffinity column had a capacity of 200 ng of aflatoxins. The detection limits (S/N = 3) for aflatoxin G1, B1, G2 and B2 were 0.03, 0.07, 0.05 and 0.09 µg·kg⁻¹, and the corresponding quantification limits (S/N = 10) were 0.10, 0.25, 0.18 and 0.30 µg·kg⁻¹, respectively. The recoveries of aflatoxins in samples were 90.1%-104.4% and RSDs were <4.4%. The developed method was further applied to the determination of aflatoxins in peanut, vegetable oil and tea samples, and the results indicated that peanut (26.9%), vegetable oils (28.0%) and tea (5.3%) samples were contaminated with aflatoxins, with levels ranging from 0.49 to 20.79 µg·kg⁻¹.

[Prelimilary Study on Manipulative Resetting Pictures of Shang ke hui zuan (Collected Compilation of Traumatology)].

Shang ke hui zuan (Collected Compilation of Traumatology) written by Hu Ting-guang, a famous doctor in the Qing Dynasty, is a comprehensive traumatology book, which records 16 manual reduction pictures with a five-word verse in each one and 9 seven-word verses in Chapter 3 supplementing the former and vice versa. Vivid pictures and brief verses matched with each other are convenient to imitate and recite, and play a vital role in promotion of inheritance and transmission of manual reduction technique on traumatology.

[Study on pictures in Waike Xinfa Yaojue].

Collecting 265 pictures, Waike Xinfa Yaojue (written in Qing Dynasty) is the extant ancient surgical book with most pictures. Pictures could be divided into four kinds, i.e. disease and symptoms, meridians, pulse taking and taboos. Pictures on disease and symptoms account for the largest number and appeared most often and those pictures are also with special subject characteristics and of great importance to the diagnosis of surgical diseases. Pictures in ancient medical books played an important role in academic heritage.

[Taking a look at the characteristics of the diagnostic methods of Traditional Chinese Medicine (TCM) from the periodical literature in the republican period of China].

Compiling 198 pieces of literature related to TCM diagnostic method in 210 kinds of medical periodicals during the republican period of China, they can be divided into three categories by different characteristics: theoretical research, popular common sense, works and lectures etc. Taking a look at the contents of the articles, there were 74 pieces of comprehensive literature (37%) 20 pieces of inspection literatures (10%) 6 pieces of auscultation and olefaction literature (3%) 11 pieces of inquiry literature (6%) 87 pieces of palpation literature (44%) Pulse diagnosis is a traditional distinguishing diagnostic method which involves much research. The literature on pulse diagnosis was in the majority The articles about theoretical research mostly explained the TCM principles with western medical knowledge and advocated diagnosing the disease with western medical apparatus, which was the characteristic of that time.