ObjectiveThe detection of RNA single nucleotide polymorphism (SNP) is of great importance due to their association with protein expression related to various diseases and drug responses. At present, splintR ligase-assisted methods are important approaches for RNA direct detection, but its specificity will be limited when the fidelity of ligases is not ideal. The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection. MethodsIn this study, a dual-competitive-padlock-probe (DCPLP) assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation. To verify the method, we employed dual competitive padlock probe-mediated rolling circle amplification (DCPLP-RCA) to genotype the CYP2C9 gene. ResultsThe specificity was well improved through the competition and strand displacement of dual padlock probe, with an 83.26% reduction in nonspecific signal. By detecting synthetic RNA samples, the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L. Furthermore, clinical samples were applied to the method to evaluate its performance, and the genotyping results were consistent with those obtained using the qPCR method. ConclusionThis study has successfully established a highly specific direct RNA SNP detection method, and provided a novel avenue for accurate identification of various types of RNAs.

This study aimed to propose an operational definition of late-onset hypogonadism (LOH) that incorporates both clinical symptoms and serum testosterone measurements to evaluate the prevalence of LOH in aging males in China. A population-based sample of 6296 men aged 40 years-79 years old was enrolled from six representative provinces in China. Serum total testosterone (TT), sex hormone-binding globulin (SHBG), and luteinizing hormone (LH) were measured and free testosterone (cFT) was calculated. The Aging Males' Symptoms (AMS) scale was used to evaluate the LOH symptoms. Finally, 5078 men were included in this analysis. The TT levels did not decrease with age (P = 0.59), and had no relationship with AMS symptoms (P = 0.87 for AMS total score, P = 0.74 for ≥ 3 sexual symptoms). The cFT levels decreased significantly with age (P < 0.01) and showed a negative association with the presence of ≥ 3 sexual symptoms (P = 0.03). The overall estimated prevalence of LOH was 7.8% (395/5078) if a cFT level <210 pmol l

Under the guidance of the theory of traditional Chinese medicine （TCM）， charcoal drugs are widely used in clinical treatment of various bleeding syndromes， in addition， they also have the effect in anti-diarrhea and anti-ulcer， but charcoal drugs are especially effective in stopping bleeding. According to the changes in the properties after processing， the hemostatic effect of charcoal drugs can be roughly divided into two categories. One is not used for hemostasis itself， but used for hemostasis after processing. The other is used for hemostasis itself， and the drug properties are changed or the hemostatic ability is enhanced after processing. By summarizing researches on historical evolution， processing mechanism and pharmacological effects of the commonly used hemostatic charcoal drugs， the author found that preservation or increase of active substances after processing was closely related to the hemostatic effect of charcoal drugs. The hemostatic mechanism mainly involves the influence of coagulation system and platelet function， etc. At the same time， combined with the theory of Qi chromatograph of TCM supramolecular， this paper puts forward the supramolecular research strategy on hemostatic mechanism of charcoal drugs， in order to provide reference for revealing the scientific connotation of charcoal drugs for hemostasis.

Zhenwutang is the classic formula of Wenyang Lishui in " Treatise on Febrile Diseases" . It consists of five kinds of medicines, namely Aconiti Lateralis Radix Praeparata, Zingiberis Rhizoma Recens, Atractylodis Macrocephalae Rhizoma, Poria, Paeoniae Radix Alba or Paeoniae Radix Rubra. It has been included in the Catalogue of Ancient Classics (The First Batch) issued by the State Administration of Traditional Chinese Medicine in 2018.There are two articles in the " Treatise on Febrile Diseases" about the syndromes of Zhenwutang. On the basis of Article 82 of the Chapter of Taiyang disease, the Taiyang disease is demonstrated by sweating, but after that the patient still suffers from fever, epigastric throb, dizziness and shiver, this shall be treated with Zhenwutang. According to Article 316 of the chapter of Shaoyin disease, Shaoyin disease lasts for two or three days, until the fourth and fifth days, and is demonstrated by abdominal pain, difficult urination, severe pain in the limbs and diarrhea, the patient have a cough, diuresis, diarrhea and vomit, this shall be treated with Zhenwutang. The original texts discuss the basic pathogenesis is edema syndrome due to Yin and Yang deficiency, with symptoms of unfavorable urination, heavy limbs or edema, pale tongue, white moss and heavy pulse as dialectical points. In terms of the compatibility of prescriptions, usage and dosage, and the interpretation of prescriptions, this paper examines Zhenwutang syndromes in " Treatise on Febrilediseases" , summarizes the material basic research of Zhenwutang, and provides the basis for the quality control of its compound pharmacodynamics. This study summarizes the progress of pharmacological research for Zhenwutang in recent years, finds that Zhenwutang has many efficacies, such as cardiotonic, diuretic, lowering lipid, anti-oxidation, improving renal function and balancing of water and liquid metabolism, lists traditional Chinese medicine(TCM) syndromes treated with Zhenwutang, such as heart palpitations, edema, cough, abdominal pain, sipping, dizziness, and Western medicine diseases, like congestive heart failure, pulmonary heart disease, bronchial asthma, chronic colitis, cirrhosis, ascites, chronic glomerulonephritis, chronic renal failure, benign prostatic hyperplasia and Meniere's syndrome, expounds the principle of Zhenwutang for " treating different diseases with the same therapy" , in order to provide useful reference for the research and development and clinical application of Zhenwutang and more classic prescriptions.

Yangjing Zhongyu Tang, from FU Qing-zhu Nvke, is a famous classical formula of clinical value for treating blood deficiency syndrome of female infertility in the Qing dynasty. The prescription seems plain, but it is rigorous and effective with high research value. In this paper, the ancient books and modern documents of Yangjing Zhongyu Tang were analyzed from the aspects of traceability of Chinese materia medica in the formula, pharmacological research, clinical research, etc. It could be concluded that the pharmacological studies of this formula were mostly to investigate the mechanism and efficacy of its treatment for infertility, but there was a lack of comprehensive interpretation of the structure, function and principle of pharmacodynamic substance in this formula. In this paper, combined with the contents of supramolecular imprinting template and network pharmacology, the new direction of pharmacological research of Chinese medicine compound was put forward. Quality control of Yangjing Zhongyu Tang mostly based on small molecule compounds, so it was suggested to break the inherent thinking, and increase the detection of macromolecule compounds and supramolecular of Chinese materia medica. The druggability of this formula involved in the preliminary study of decoctions, plasters and granules. Considering the quality stability of Chinese materia medica in the formula and the scientificity of druggability of this formula, the compatibility principle of traditional Chinese medicine (TCM) and modern supramolecular chemistry theory could be combined to study the change law of druggability of TCM before and after compatibility, so as to provide new reference materials for the follow-up clinical application and development of Yangjing Zhongyu Tang.

This paper discussed the processing methods of Pinelliae Rhizoma in the successive dynasties by consulting ancient and modern books, literature and related codes. The processing methods of Pinelliae Rhizoma has experienced various historical periods. Since the processing method of Pinelliae Rhizoma processed by slicing appeared in the Huangdi Neijing, there have been a series of different processing methods and requirements are founded, such as cleansing, cutting, decocting and so on. Among them, the raw Pinelliae Rhizoma, Pinelliae Rhizoma Praeparatum, Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine, Pinelliae Rhizoma Praeparatum cum Alumine are still in use today and are widely used. However, there are differences in the processing methods, accessories and their dosages of these various processed products in different dynasty. The processing technology and quality standard of Pinelliae Rhizoma need to be further standardization.

Objective: Based on the analysis of the total components of fingerprint and the determination methods of the existing components in the 2015 edition of Chinese Pharmacopoeia, the "point-line-surface" quality standard of Yangjing Zhongyu Tang was established by the "point" of each single component (morroniside, loganin, paeoniflorin, ferulic acid and verbascoside) to the "line" of multicomponent and the "face" of fingerprint of the whole component. Method: XB-C₁₈ column (4.6 mm×250 mm, 5 μm) was used for gradient elution of 0.1% phosphoric acid aqueous solution-acetonitrile. The column temperature was 30℃, the injection volume was 10 μL, the flow rate was 1.0 mL·min^-1, and the detection wavelengths were 240, 316, 230, 334 nm. The contents of these five components in Yangjing Zhongyu Tang were determined by three correction methods, external standard method and regression equation method. At the same time, the fingerprint of Yangjing Zhongyu Tang were analyzed by total component analysis and similarity evaluation. Result: With ferulic acid as reference, the relative correction factor (f) of morroniside, loganin, paeoniflorin and verbascoside were 0.392 1, 0.421 4, 0.261 7, 0.268 6 by multi-point correction method, and their f (slope correction method) were 0.385 4, 0.419 4, 0.255 9, 0.274 0, respectively. Twenty characteristic peaks of fingerprint were analyzed and the similarity was ≥ 0.999.There was no significant difference in the contents of these five components from Yangjing Zhongyu Tang determined by the quantitative assay of multi-components by single-marker (QAMS) correction method, the external standard method and the regression equation method. Conclusion: The comprehensive quality standard established by the total component analysis of fingerprint combined with various determination methods of existing components in the 2015 edition of Chinese Pharmacopoeia has been validated in famous classical formula of Yangjing Zhongyu Tang, which can provide ideas and methods for the quality control with quantitative determination and fingerprint of other famous classical formulas.

Based on the characteristics of biological supramolecules and the law of " imprinting template",the research status and common problems in " maintaining medicinal properties after carbonisatus" in traditional Chinese medicine( TCM) were analyzed,and the further countermeasures were put forward. According to the historical evolution of " maintaining medicinal properties after carbonisatus" in TCM processing,the origin of its common problems was clarified by using the theory of biosupramolecular chemistry. TCM is a megacomplex biological supramolecular system,so TCM processing is just the processing of megacomplex biological supramolecular system,and its essence is a TCM pharmaceutical technology with chemical changes in host and guest of biological supramolecular system with or without adjuvant material under high temperature and humidity. In this study on pharmaceutical technology,host molecule was destructed in the process of carbonizing,but guest molecule was retained. The changing law of the host and guest molecule was controlled by the " imprinting template",which was reflected in the degree of change in the drug properties and efficacy of the decoction pieces. Supramolecular chemistry ran through the whole process,and the " imprinting template" of charcoal medicine was characterized by the supramolecular topological structure characteristics and imprinting behavior. After being combined with the quantitative mathematical model of heating degree in processing,it can realize the accurate processing of " maintaining medicinal properties after carbonisatus" from the source,quantitatively control the quality of carbonic herbs,and formulate stable and controllable quality standards.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Records ; Reference Standards ; Technology

More and more disputes have happened to confront us continuously since the separation of Japanese(wild) honeysuckle flower in Chinese Pharmacopoeia in 2005. The state pharmacopeia committee decided to separate Japanese(wild) honeysuckle flower into two species for japanese(wild) honeysuckle flower, but didn't define their the convincing reasons still did not provide to us as a result that two medicines are not described the differences in natural properties, efficiency and indication, usage and dosage, as well as not given a resolving methodand specific solution. It was known for us that in the history of traditional Chinese medicine（TCM）, the phenomenenphenomena of the "one drug from multi-species" and "one species for multi-drug" are very ordinary thingswere ubiquitous. Whether separation of the drug species are separated shall be decided to by clinical efficiency. Through Chinese pharmacopoeia (2015 edition) issue of Chinese Pharmacopoeia the 2015 issue of the Chinese Pharmacopoeia, we still cannot find a scientific solution for the dispute of for Japanese (wild) honeysuckle flower, perhaps because of insufficient reorganization of TCM clinical medication regularities, such as "treatment of different diseases with same drug", " treatment of the same disease with different drugs" and "treatment of the same syndrome with multi-prescriptions", and "one prescription treating multiple syndromes"; lack of in-depth analysis for multi-component TCM compounds and autonomisation of "supermolecular template" for organs and meridians; less attention to the advance of efficacy and safety evaluation technologies for multi-component TCM compounds; impacts from the medication mode of "one ingredient-one composition-one effect"; as well as insufficient research methods for bioequivalent evaluation in preclinic and clinic studies . The dispute for species combination or separation for Japanese(wild) honeysuckle flower was apparently caused by regional economy, drug biological equivalent of efficacy and safety, but arising from clinical principles for systematical syndrome treatment with TCM, or concepts in the treatment of diseases with TCM or western medicines. This paper focused on current studies on Japanese(wild) honeysuckle flower in the combination with TCM clinical medication regularities, such as "treatment of different diseases with same drug", "treatment of the same disease with different drugs" and "treatment of the same syndrome with multi-prescriptions", and "one prescription treating multiple syndromes", expounded the specific pharmacological regularity of "heterogeneous equivalence" of Japanese (wild) honeysuckle flower, and put forward methods for studying bioequivalence of Japanese(wild) honeysuckle flower, in order to solve the combination and separation of Japanese(wild) honeysuckle flower and lay a foundation for promoting the development of Chinese herbal medicine industry.

Controlled sustained-release preparation (CSRP) is a definitely important source of innovative medicines, however, its further researches and development are limited due to the complexity of Chinese medicine (CM) compositions, and a series of unclear key problems like the apparent properties from physical and chemical aspects. Combined with the CM characteristics of the multiple components and synergistic effect, a series of bottleneck problems in the preparation of CSRP were analyzed. Then the mechanism and influences of supermolecular chemical theory in the CSRP about the multiple-components overall-control were explored under the guideline of "overall control, synchronous dissolution" in CSRP, based on the previous explications about supramolecular chemistry theories and with the application of single component precision controlling technologies. The supermolecular mechanism of multi-component overall control and synchronous release of the CSRP was investigated, providing an useful academic reference for the research and development of the CSRP and laying the theoretical and technical foundation for the manufacturing process of CSRP.