Transcriptome-based Screening and Validation of Key Enzyme Genes for Polygonatum Polysaccharide Metabolism / 中国实验方剂学杂志

ObjectiveTo screen and validate key enzyme genes affecting the polysaccharide content in different Polygonatum species and perform in-depth amino acid sequence analysis by transcriptomic analysis of P. zanlanscianense， P. kingianum， and P. cyrtonema rhizomes to enrich the transcriptome data of Polygonatum plants and provide references for polysaccharide biosynthesis mechanism and genetic improvement. MethodThe Polygonatum transcriptome was sequenced and analyzed using the Illumina NovaSeq high-throughput sequencing platform， and the differences in the transcriptomes of the three Polygonatum species were compared and according to the annotations of Nr， Gene Ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） databases. The key enzymes in the polysaccharide metabolism pathway were screened， and the expression of key enzyme genes was clustered and correlated with the polysaccharide content. Finally， Real-time polymerase chain reaction （Real-time PCR） was performed to validate the eight key enzyme genes， and the key genes of polysaccharide biosynthesis were further screened for homologous gene sequence analysis in combination with sequencing results， followed by constructing phylogenetic trees， predicting motifs， conserved structural domains， protein sequence isoelectric points， and molecular weights， and constructing 3D protein structures by using homology modeling method. ResultThe annotation of the Nr database revealed that three Polygonatum species had the highest gene homology with Asparagus officinalis. GO database annotation results showed that three Polygonatum species differed significantly in binding， catalytic activity， metabolic processes， and cellular components， while the KEGG pathway annotation results indicated that three Polygonatum species differed significantly in the starch and sucrose metabolic pathway and galactose metabolic pathway. According to clustering analysis， correlation analysis， Real-time PCR， expression profiles， and structural and functional predictions of amino acid sequences， the key enzyme significantly affecting the polysaccharide content in different Polygonatum species was inferred to be β-fructofuranosidase （sacA）. ConclusionSacA may be the main influencing factor for the difference in polysaccharide content of Polygonatum， and is also an important reason why Polygonatum polysaccharides are mainly fructans.

Genomic, transcriptomic, and epigenomic analysis of a medicinal snake, Bungarus multicinctus, to provides insights into the origin of Elapidae neurotoxins

The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.

Comparison of the Anti-inflammation,Analgesic Effects of Decoctions Extracted from Aconiti Lateralis with Different Leaf Shapes from Different Producing Areas / 中国药房

OBJECTIVE:To compare the anti-inflammation,analgesia effects of decoctions extracted from Aconiti lateralis with different leaf shapes(dahua leaf,xiaohua leaf)from different producing areas(Jiangyou,Butuo). METHODS:Animals were randomly divided into blank group(distilled water),positive group,groups of Aconiti lateralis with dahua,xiaohua leaf from Ji-angyou,groups of Aconiti lateralis with dahua,xiaohua leaf from Butuo(with dose of 5 g/kg,calculated by crude drug). The an-ti-inflammation effect of decoctions extracted from Aconiti lateralis with different variety sources and leaf shapes was investigated by xylene-induced ear swelling test (n=12) in mice and egg white-induced toe swelling test (n=10) in rats (positive drug was Dexamethasone acetate tablet,0.005 g/kg). And its analgesic effect was investigated by acetic acid-induced writhing body reaction test(n=12)and hot-plate-induced pain test(n=12)in mice(positive drug was Morphine hydrochloride tablet,0.0025 g/kg). RE-SULTS:The decoctions extracted from Aconiti lateralis with dahua,xiaohua leaf from Butuo and xiaohua leaf from Jiangyou can significantly reduce the ear swelling degree(P<0.01). The decoctions extracted from Aconiti lateralis with dahua leaf from Jiangy-ou and Butuo can significantly decrease the toe swelling degree after 6 h of medication(P<0.05). And decoctions extracted from Aconiti lateralis with xiaohua leaf from Butuo can significantly reduce the number of writhing body in mice with acetic acid-in-duced pain and prolong the pain threshold of mice with hot-plate-induced pain (P<0.05 or P<0.01). CONCLUSIONS:Aconiti lateralis with dahua and xiaohua leaf from Butuo and with xiaohua leaf from Jiangyou show better anti-inflammation effect,and Aconiti lateralis with xiaohua leaf from Butuo shows better analgesic effect.

Influencing factors of knee function in osteoarthritis patients after total knee arthroplasty / 中国组织工程研究

BACKGROUND:Functional recovery after joint arthroplasty is an important indicator to evaluate the effect and prognosis of total knee arthroplasty, and is affected by patients and many kinds of outside factors. OBJECTIVE:To investigate the influencing factors of functional recovery of knee in patients with knee osteoarthritis after total knee arthroplasty. METHODS:Clinical data of 76 osteoarthritis patients undergoing total knee arthroplasty in the Dongying People’s Hospital from August 2007 to August 2013 were retrospectively analyzed. Hospital for Special Surgery knee score, knee pain Visual Analogue Score, range of motion and posterior slop angle of tibial plateau were compared before and 6 months after surgery. Changes in anterior condylar offset were recorded before and after femoral prosthesis arthroplasty. SPSS 21.0 software was used to analyze the influential factors for Hospital for Special Surgery knee scores after arthroplasty. RESULTS AND CONCLUSION:(1) Hospital for Special Surgery knee score, knee pain Visual Analogue Score, and range of motion were significantly higher 6 months after arthroplasty than that before arthroplasty (P<0.05). (2) Pearson correlation analysis demonstrated that preoperative Hospital for Special Surgery knee score, preoperative knee pain Visual Analogue Score, preoperative range of motion, preoperative posterior slop angle of tibial plateau and preoperative and postoperative anterior condylar offset changes were positively correlated with postoperative Hospital for Special Surgery knee score (P<0.05). Body mass index and inversion deformity angle were negatively correlated with postoperative Hospital for Special Surgery knee score (P<0.05). (3) Multiple linear regression analysis showed body mass index was negatively correlated with postoperative Hospital for Special Surgery knee score (P<0.05). Preoperative Hospital for Special Surgery knee score, preoperative knee pain Visual Analogue Score and preoperative posterior slop angle of tibial plateau were positively correlated with postoperative Hospital for Special Surgery knee score (P<0.05). (4) These results indicated that total knee arthroplasty significantly improved knee joint function. Body mass index, preoperative Hospital for Special Surgery knee score, preoperative knee pain Visual Analogue Score, preoperative posterior slop angle of tibial plateau and preoperative and postoperative anterior condylar offset changes were correlated with the recovery of joint function after total knee arthroplasty.