italic>O-methyltransferases (OMTs) are one of the key tailoring enzymes in the biosynthesis of many natural products, O-methylation can not only reduce the reactivity of natural products, but also alter their solubility, stability and biological activities. Based on the transcriptome data of Ardisia japonica, a full-length cDNA sequence of candidate OMT (termed as AjOMT1) was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli (E. coli) for the first time. In vitro enzyme catalytic activity assay showed that the recombinant AjOMT1 could effectively catalyze quercetin to form O-methylated products. Most importantly, AjOMT1 showed unprecedented substrate promiscuity towards structurally various compounds including flavonoids, stilbenes, coumarins, alkaloids and phenylpropanoids, especially preferring to the compounds with adjacent phenolic hydroxyl groups, and showed regio-selectivity. These results suggested that AjOMT1 could be used as the tool enzyme to conduct O-methylation of different types of compounds to produce diverse active methylated products, and provide a new method for drug discovery, even universal part for synthetic biology of natural products.

【Objective】 To explore the correlation between CSAG1 expression and the prognosis and tumor-infiltrating lymphocytes in renal clear cell carcinoma (RCCC), and to predict the survival and tumor progression. 【Methods】 The gene expression profiles and clinical information of CSAG1 were downloaded from the Cancer Genome Atlas (TCGA). Based on the differential mRNA expression, GO annotation and KEGG pathway analysis were performed. The relationship between CSAG1 and tumor immune infiltration was assessed with Tumor Immunoassay Resource (Timer 2.0) database. The mRNA expression of CSAG1 in human RCCC specimens was validated with qRT-PCR. 【Results】 CSAG1 expression was significantly higher in RCCC tissues than in normal tissues (P<0.05). The qRT-PCR results revealed that the mRNA level of CSAG1 was consistent with that predicted by bioinformatic analysis. The KEGG analysis and GO annotation indicated high GSAG1 expression in RCCC was related to transmembrane transport, tricarboxylic acid cycle and lysosome. CSAG1 expression was positively related to the infiltration of pDC, aDC, CD8⁺ T cells, cytotoxic cells, TFH, TH1 cells, Tem, NK CD56^dm cells, Treg and T cells, but negatively correlated with macrophage infiltration. 【Conclusion】 CSAG1 may be associated with poor prognosis of RCCC and become a potential immunotherapy target.

OBJECTIVES: To investigate the association of single nucleotide polymorphisms (SNPs) of myeloid differentiation factor 88 (MyD88) and Toll-like receptor adaptor molecule 1 (TICAM1) and their interactions with community-acquired pneumonia (CAP) in children. METHODS: Improved multiple ligase detection reaction assay was used for detecting the polymorphisms of nine tagging SNPs of the MyD88 and TICAM1 genes in 375 children with CAP who attended the Department of Pediatrics of the Second Affiliated Hospital of Yan'an University Medical School from August 2015 to September 2017 and 306 healthy children who underwent physical examination. A logistic regression analysis was used to evaluate the association between the distribution of genotypes and their interactions with CAP in children. RESULTS: The polymorphism of the TICAM1 gene at rs11466711T/C locus was closely associated with the susceptibility to CAP in children (P<0.05). The AA genotype of rs35747610G/A locus significantly reduced risk of sepsis in children with CAP (P<0.05). The AA genotype of rs6510826G/A locus was significantly associated with the increase in C-reactive protein level in children with CAP (P<0.05). The GG genotype of the MyD88 gene at rs7744A/G locus significantly increased the risk of respiratory failure and circulatory failure (P<0.05). The multiplicative interactions between MyD88 gene rs7744A/G and TICAM1 gene rs11466711T/C, rs2292151G/A, rs35299700C/T, and rs35747610G/A loci were significantly associated with the susceptibility to CAP, the severity of CAP, and the risk of sepsis in children (P<0.05). CONCLUSIONS The gene polymorphisms of MyD88 and TICAM1 and their interactions are closely associated with CAP in children, with a synergistic effect on the development and progression of CAP in children.
Child ; Humans ; Adaptor Proteins, Vesicular Transport/genetics* ; Community-Acquired Infections/genetics* ; Myeloid Differentiation Factor 88/genetics* ; Pneumonia/genetics* ; Polymorphism, Single Nucleotide ; Sepsis

OBJECTIVE: To assess the practical and health economical values of non-invasive prenatal test (NIPT) in Changsha Municipal Public Welfare Program. METHODS: A retrospective analysis was carried out on 149 165 women undergoing NIPT test from April 9, 2018 to December 31, 2019. For pregnant women with high risks, invasive prenatal diagnosis and follow-up of pregnancy outcome were conducted. The cost-benefit of NIPT for Down syndrome was analyzed. RESULTS: NIPT was carried out for 149 165 pregnant women and succeeded in 148 749 cases (99.72%), for which outcome were available in 148 538 (99.86%). 90% of pregnant women from the region accepted the screening with NIPT. 415 (0.27%) were diagnosed as high risk. Among these, 381 (91.81%) accepted amniocentesis, which led to the diagnosis of 212 cases of trisomy 21 (PPV=85.14%), 41 cases with trisomy 18 (PPV=48.81%) and 10 cases with trisomy 13 (PPV=20.83%). The sensitivity and specificity of NIPT for trisomy 21, trisomy 18 and trisomy 13 were (97.70%, 99.98%), (97.62%, 9.97%) and (100%, 99.97%), respectively. In addition, 213 and 30 cases were diagnosed with sex chromosomal aneuploidies (PPV=46.2%) and other autosomal anomalies (PPV=16.57%), respectively. For Down syndrome screening, the cost and benefit of the project was 120.79 million yuan and 1,056.95 million yuan, respectively. The cost-benefit ratio was 1: 8.75, and safety index was 0.0035. CONCLUSION NIPT is a highly accurate screening test for trisomy 21, which was followed by trisomy 18 and sex chromosomal aneuploidies, while it was less accurate for other autosomal aneuploidies. The application of NIPT screening has a high health economical value.
Aneuploidy ; Cost-Benefit Analysis ; Female ; Humans ; Noninvasive Prenatal Testing ; Pregnancy ; Retrospective Studies ; Trisomy 18 Syndrome/genetics*

Molecular pharmacognosy is a science of classification and identification, cultivation and protection, and production of active ingredients of graduated drugs at the molecular level. The proposal of molecular pharmacognosy allows the research of crude drugs to advance from the microscopic level to the genetic level. Pueraria lobata root, as a medicinal and edible plant, has high application value and economic value. There are many varieties that are easy to cause confusion, and it is not easy to distinguish and identify according to traditional identification methods. Moreover, the research of P. lobate root at the genetic level is still relatively shallow. the study received extensive attention of scholars. This article reviews recent research on molecular identification of P. lobate, transcriptome sequencing, cloning and synthesis of functional genes of P. lobate root in recent years in order to provide references for further promoting the development and utilization of P. lobate root and its active ingredients.
Pharmacognosy ; Plant Roots/genetics* ; Pueraria

Objective:To compare the chemical constituents of Puerariae Flos from three different varieties of Pueraria montana var. lobata， P. montana var. thomsonii and P. montana var. montana. Method:Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS） was used with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-20 min， 10%-30%B； 20-30 min， 30%-55%B； 30-35 min， 55%-95%B； 35-37 min， 95%B； 37-40 min， 95%-10%B）， the flow rate was 0.25 mL·min^-1. Electrospray ionization （ESI） was used to scan and collect MS data in positive and negative ion modes with scanning range of m/z 50-1 500. The chemical components from different sources of Puerariae Flos were identified in combination with the chemical composition database and literature information. After the obtained data were normalized by MarkerView^TM 1.2.1， they were imported into SICMA-P 14.1 software for principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） to select the main differentiated components among the three different varieties. Result:A total of 35 compounds were identified from three different varieties of Puerariae Flos， including 22 isoflavones， 6 flavonoids and 7 saponins. The flowers of P. lobata， P. montana var. thomsonii and P. montana var. montana contained 32， 35， 33 compounds， respectively. And 18 differential compounds were screened under the positive and negative ion modes， including kakkalide， tectoridin， 6″-O-xylosyl-tectoridin， 4'-methyltectorigenin-7-glucoside， glycitin， 6″-O-xylosyl-glycitin， irisolidone， kaikasaponin Ⅲ， 6″-O-malonylglycitin， kakkalidone， tectorigenin， rutin， soyasaponin BB， vitexin， biochanin A， genistin， kakkatin， azukisaponin Ⅱ. Conclusion:This research is the first to systematically study the chemical constituents of the flower of P. montana var. montana， although the flower of P. montana var. montana is used as adulterants， it has high contents of tectoridin and 6″-O-xylosyl-tectoridin， which has great potential for development. The efficacy components such as kakkalide and tectoridin in Puerariae Flos from the three sources of varieties are obviously different， and it is necessary to carefully consider the application of these three varieties as Puerariae Flos.

The pathogenesis of metabolic syndrome （MS） includes insulin resistance （IR）， central obesity， chronic low-grade inflammation， oxidative stress， endoplasmic reticulum stress， elevated free fatty acid levels， intestinal flora imbalance， renin angiotensin system abnormality， and autophagy activity deficiency， etc. Most researchers believe that IR plays a central role in the pathogenesis of MS， and abdominal obesity is an important initial factor of MS. According to the incidence and clinical characteristics， MS is classified as "obesity" "pidan" " abdominal fullness " and other diseases. It is said that the pathogenesis of MS is related to the deficiency of spleen and kidney， the formation of phlegm， turbidity， blood stasis and other pathological products， which damage the body's functions of qi， blood， yin and yang. Traditional Chinese medicine （TCM） has unique advantages in treating MS based on the holistic view and syndrome differentiation concept. It has multi-level， multi-target and multi-channel treatment characteristics. It can intervene insulin signal transduction， regulate adipocyte factor secretion level， relieve oxidative stress and endoplasmic reticulum stress response， regulate intestinal flora and renin angiotensin system， reduce free fatty acid level and regulation Autophagy and other ways to improve chronic low-grade inflammation and IR status， and then comprehensive prevention and treatment of MS and its complications. However， the following problems still exist：lack of high-quality randomized controlled clinical research and large sample real-world research， clinical unified diagnosis and treatment standard has not yet formed， lack of genetic animal model in basic research， relatively single signal pathway and target of experimental research， and difficulty in timely formation of clinical transformation of scientific research achievements. Therefore， we should make full use of modern scientific and technological means to carry out systematic and standardized multicenter， large sample， high-quality randomized controlled trials or real-world research， we should prepare perfect animal models， focus on the crosstalk relationship between multiple related cell signaling pathways， and actively explore the potential relationship between signaling pathways and prescription compatibility， so as to actively promote basic scientific research achievements Clinical practice may be the key research direction in the prevention and treatment of MS in TCM.

Metabolic syndrome (MS) is a pathological condition characterized by central obesity, insulin resistance, hypertension, and hyperlipidemia. With the increase of poor dietary habits and lifestyles in modern society, especially the poor living habits of sedentariness and less movement, the prevalence of MS has increased year by year. According to relevant data, the number of MS patients worldwide will reach about 2.568 billion by 2040, which will seriously endanger human life and health. Huanglian Wendantang, as a famous traditional Chinese medicine prescription for clearing away heat and drying dampness, regulating Qi and resolving phlegm, and benefiting the stomach and gall, has been proved to have significant pharmacological effects in lowering blood fat, reducing blood sugar and resisting inflammation by modern pharmacological studies, and widely used in the treatment of metabolic diseases, cardiovascular diseases and other systemic diseases. In recent years, a large number of studies have proved that Huanglian Wendantang has a significant effect on MS. In terms of clinical efficacy, it could significantly improve the pathological state of obesity, dyslipidemia, abnormal glucose metabolism and hypertension in MS patients. Meanwhile, it could also interfere with the inflammatory state, prethrombotic state, abnormal vascular regulation and other potential risk factors in the body, with a high safety and fewer side effects. In terms of experimental study, it could enhance the insulin sensitivity, and improve the insulin resistance of MS animal models and cell models through interventions in insulin signal transduction, inflammatory response, and antioxidant stress. By retrieving PubMed, CNKI, Weipu, Wanfang and other databases, the author summarized the study reports of Huanglian Wendantang on MS in recent years in three aspects: theoretical study, clinical efficacy study and experimental mechanism study, in the expectation of provide some scientific references for in-depth study of the mechanism of Huanglian Wendantang in treating MS and the development and clinical promotion of the prescription.

Objective:To preliminarily interpret the compatibility of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma in chemical and pharmacodynamic levels，and provide theoretical basis for its clinical application.Method:Rapid resolution liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry（RRLC-Q-TOF-MS） was applied to identify and analyze the changes in chemical components of the Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma before and after compatibility. The anti-fatigue activity before and after compatibility of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma was detected by weight-loading swimming experiment and determination of levels of serum urea，blood lactic acid and hepatic glycogen.Result:A total of 51 compounds were identified in mixture decoction of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma. Malonyl ginsenoside mRg₁，mRb₁，mRb₂，mRb₃ and mRd contents were significantly decreased，while ginsenoside Rb₁，Rb₂，Rb₃，Rd，F₂ and Rg₃ contents were significantly increased in the compatibility mixture. According to pharmacodynamics study，as compared with those in the blank control group，swimming time of mice was significantly prolonged in all other groups （P<0.01），serum urea nitrogen（P<0.05，P<0.01） and lactic acid（P<0.05，P<0.001） levels of mice in the combined decoction and the single decoction groups were significantly lowered，while liver glycogen levels were significantly elevated（P<0.05，P<0.01）. The anti-fatigue ability of the combined decoction of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma was higher than that of the single decoctions.Conclusion:In this article, the effect enhancing mechanism of compatibility of Ginseng Radix et Rhizoma Rubra with Rhodiolae Crenulate Radix et Rhizoma was revealed based on the chemical changes, providing theoretical reference for the clinical application and development of products.