Electroacupuncture alleviates orofacial allodynia and anxiety-like behaviors by regulating synaptic plasticity of the CA1 hippocampal region in a mouse model of trigeminal neuralgia.

Objective: Trigeminal neuralgia (TN), one of the most severe and debilitating chronic pain conditions, is often accompanied by mood disorders, such as anxiety and depression. Electroacupuncture (EA) is a characteristic therapy of Traditional Chinese Medicine with analgesic and anxiolytic effects. This study aimed to investigate whether EA ameliorates abnormal TN orofacial pain and anxiety-like behavior by altering synaptic plasticity in the hippocampus CA1. Materials and methods: A mouse infraorbital nerve transection model (pT-ION) of neuropathic pain was established, and EA or sham EA was used to treat ipsilateral acupuncture points (GV20-Baihui and ST7-Xiaguan). Golgi-Cox staining and transmission electron microscopy (TEM) were administrated to observe the changes of synaptic plasticity in the hippocampus CA1. Results: Stable and persistent orofacial allodynia and anxiety-like behaviors induced by pT-ION were related to changes in hippocampal synaptic plasticity. Golgi stainings showed a decrease in the density of dendritic spines, especially mushroom-type dendritic spines, in hippocampal CA1 neurons of pT-ION mice. TEM results showed that the density of synapses, membrane thickness of the postsynaptic density, and length of the synaptic active zone were decreased, whereas the width of the synaptic cleft was increased in pT-ION mice. EA attenuated pT-ION-induced orofacial allodynia and anxiety-like behaviors and effectively reversed the abnormal changes in dendritic spines and synapse of the hippocampal CA1 region. Conclusion: EA modulates synaptic plasticity of hippocampal CA1 neurons, thereby reducing abnormal orofacial pain and anxiety-like behavior. This provides evidence for a TN treatment strategy.

[Professor ZHENG Liang's experience and understanding of postoperative facial paralysis treated with acupotomy].

Professor ZHENG Liang believes that the main pathogenesis of postoperative facial paralysis is related to the retarded circulation of qi and blood and malnutrition of tendons and vessels in local area because of local retention of "stasis" after surgical trauma. In treatment of postoperative facial paralysis with acupotomy, the abnormal facial structure after operation should be considered specially. The region where acupotomy is exerted is determined by taking surgical scar as the center so that the local adhesion can be released and separated. In treatment, the knife needle should be as fine as possible and the attention be paid to the direction of needle insertion and the release amplitude. The frequency of acupotomy should be once per week.

[DNA barcoding identification of original plants of a rare medicinal material Resina Draconis and related Dracaena species].

Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.

Genetic Diversity Assessment of a Chinese Medicinal Endemic Species, Aspidopterys obcordata var. obcordata, by Combined Molecular Marker Methods (ISSR & SRAP).

Aspidopterys obcordata var. obcordata, a medicinal plant endemic to China, is a narrowly distributed species and wild resources are extremely limited. To evaluate the genetic variability and degree of genetic divergence of A. obcordata var. obcordata, and to make rational scientific decisions on its harvest and germplasm conservation, we collected 122 samples from across nearly all of its distribution area and studied genetic diversity using inter-simple sequence repeats (ISSRs), sequence-related amplified polymorphisms (SRAPs), and a method combining the two techniques. The results revealed the high genetic diversity of A. obcordata var. obcordata, mainly due to its intra-population diversity, and the top two populations with the highest levels of intra-population diversity were ML and DH, individuals of which can serve as excellent germplasm candidates during the processing of germplasm screening and conservation. In general, the combining method was prior to the ISSR analyses and SRAP analyses results, except for a slight difference in the genetic structure of individual populations. Therefore, we suggest that a combination analysis of the two marker methods is ideal for evaluating the genetic diversity and genetic relationships of A. obcordata var. obcordata.

Taraxacum mongolicum protects against Staphylococcus aureus-infected mastitis by exerting anti-inflammatory role via TLR2-NF-κB/MAPKs pathways in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: As a traditional Chinese medicine, Taraxacum mongolicum has been widely used for the prevention and treatment of a variety of inflammatory and infectious diseases, and also clinically used as a remedy for mastitis. However, the scientific rationale and mechanism behind its use on mastitis in vivo are still unclear. AIM OF THE STUDY: This study aimed to investigate the protective effect and potential mechanism of Taraxacum mongolicum Hand.-Mazz. (T. mongolicum) on mastitis infected by Staphylococcus aureus (S. aureus). MATERIALS AND METHODS: Female ICR mice were given intragastrically 2.5, 5 and 10 g/kg of T. mongolicum extract twice per day for 6 consecutive days, and infected with S. aureus via teat canal to induce mastitis. Pro-inflammatory cytokine tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) levels were determined by ELISA. Myeloperoxidase (MPO) activity and distribution were measured by reagent kit and immunohistochemistry. Histopathological changes of mammary gland tissues were observed by H&E staining. Toll-like receptor 2 (TLR2) expression, phosphorylations of related proteins in nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) signaling pathways were detected by western blot. RESULTS: T. mongolicum decreased TNF-α, IL-6 and IL-1ß levels, and reduced MPO activity and distribution in sera and mammary glands with S. aureus-infected mastitis. In addition, T. mongolicum effectively attenuated histopathological damages and cell necrosis of mammary gland tissues infected by S. aureus. Moreover, T. mongolicum inhibited the expression of TLR2, and the phosphorylations of inhibitor κBα (IκBα), p65, p38, extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) proteins in mammary glands with S. aureus-infected mastitis. CONCLUSIONS: This study suggests that T. mongolicum protects against S. aureus-infected mastitis by exerting anti-inflammatory role, which is attributed to the inhibition of TLR2-NF-κB/MAPKs pathways.

[Professor YANG Wen-hui's experience in treatment of alopecia areata with the combination of acupuncture and Chinese herbs based on yingwei theory].

Professor YANG Wen-hui 's clinical experience of treating alopecia areata with combination of acupuncture and Chinese herbs based on yingwei theory was summarized. Professor YANG divides alopecia areata into ying qi disease and wei qi disease. Alopecia areata belonging to wei qi disease locates on the skin and is related to the lung and kidney, and belonging to ying qi disease locates in the meridians and is related to the heart, spleen and stomach. Acupuncture is chosen when the disease locates in the skin and meridians. If both viscera and meridians suffered, acupuncture and Chinese herbs should be combined. As for the acupuncture, professor YANG used the method of harmonizing yingwei needling to treat alopecia areata, which includes regulating the origin of yingwei, regulating the deficiency and excess of yingwei and regulating the operation of local yingwei. As for the Chinese herbs, wei qi disease can be washed out by Chinese herbs of relieving exterior wind and activating meridians and collaterals. The ying qi disease can be washed out by Chinese herbs in mild condition, and long course of disease can be given Guizhi decoction as the base, add or subtract according to syndrome.

Identification of medicinal plants within the Apocynaceae family using ITS2 and psbA-trnH barcodes.

To ensure the safety of medications, it is vital to accurately authenticate species of the Apocynaceae family, which is rich in poisonous medicinal plants. We identified Apocynaceae species by using nuclear internal transcribed spacer 2 (ITS2) and psbA-trnH based on experimental data. The identification ability of ITS2 and psbA-trnH was assessed using specific genetic divergence, BLAST1, and neighbor-joining trees. For DNA barcoding, ITS2 and psbA-trnH regions of 122 plant samples of 31 species from 19 genera in the Apocynaceae family were amplified. The PCR amplification for ITS2 and psbA-trnH sequences was 100%. The sequencing success rates for ITS2 and psbA-trnH sequences were 81% and 61%, respectively. Additional data involved 53 sequences of the ITS2 region and 38 sequences of the psbA-trnH region were downloaded from GenBank. Moreover, the analysis showed that the inter-specific divergence of Apocynaceae species was greater than its intra-specific variations. The results indicated that, using the BLAST1 method, ITS2 showed a high identification efficiency of 97% and 100% of the samples at the species and genus levels, respectively, via BLAST1, and psbA-trnH successfully identified 95% and 100% of the samples at the species and genus levels, respectively. The barcode combination of ITS2/psbA-trnH successfully identified 98% and 100% of samples at the species and genus levels, respectively. Subsequently, the neighbor joining tree method also showed that barcode ITS2 and psbA-trnH could distinguish among the species within the Apocynaceae family. ITS2 is a core barcode and psbA-trnH is a supplementary barcode for identifying species in the Apocynaceae family. These results will help to improve DNA barcoding reference databases for herbal drugs and other herbal raw materials.

[Usage of ethnomedicine on COVID-19 in China].

In December 2019, an outbreak of viral pneumonia began in Wuhan, Hubei Province, which caused the spread of infectious pneumonia to a certain extent in China and neighboring countries and regions, and triggered the epidemic crisis. The coronavirus disease 2019(COVID-19) is an acute respiratory infectious disease listed as a B infectious disease, which is managed according to standards for A infectious disease. Traditional Chinese medicine and integrated traditional Chinese and Western medicine have played an active role in the prevention and control of this epidemic. China's ethnomedicine has recognized infectious diseases since ancient times, and formed a medical system including theory, therapies, formula and herbal medicines for such diseases. Since the outbreak of the COVID-19 epidemic, Tibet Autonomous Region, Qinghai Province, Inner Mongolia Autonomous Region, Xinjiang Uygur Autonomous Region and Chuxiong Autonomous Prefecture of Yunnan, Qiandongnan Autonomous Prefecture of Guizhou have issued the prevention and control programs for COVID-19 using Tibetan, Mongolian, Uygur, Yi and Miao medicines. These programs reflect the wisdom of ethnomedicine in preventing and treating diseases, which have successfully extracted prescriptions and preventive measures for the outbreak of the epidemic from their own medical theories and traditional experiences. In this paper, we summarized and explained the prescriptions and medicinal materials of ethnomedicine in these programs, and the origin of Tibetan medicine prescriptions and Mongolian medicine prescriptions in ancient books were studied. These become the common characteristics of medical prevention and treatment programs for ethnomedicine to formulate therapeutic programs under the guidance of traditional medicine theories, recommend prescriptions and prevention and treatment methods with characteristics of ethnomedicine, and focus on the conve-nience and standardization. However, strengthening the support of science and technology and the popularization to the public, and improving the participation of ethnomedicine in national public health services and the capacity-building to deal with sudden and critical diseases are key contents in the development of ethnomedicine in the future.

The effects of genetic variation and environmental factors on rhynchophylline and isorhynchophylline in Uncaria macrophylla Wall. from different populations in China.

Uncaria macrophylla Wall. is an important Chinese medicinal herb. Rhynchophylline (RIN) and isorhynchophylline (IRN) are its major active compounds. We investigated the influence of genetic differentiation and environmental factors on the RIN and IRN to find the main influencing factors of their contents and lay the foundation for the following cultivation and breeding. We used inter-simple sequence repeat (ISSR) markers to investigate the genetic diversity, and high-performance liquid chromatography (HPLC) to measure the contents of RIN and IRN in 200 samples of U. macrophylla obtained from nine natural populations, and then to analyze the correlation between genetic differentiation, environmental factors of sampling sites and the contents of RIN and IRN. We found that High intra-population (80.05%) and low inter-population (19.95%) genetic diversity existed in the samples of U. macrophylla. To some extent, genetic differentiation and the contents of RIN and IRN had correlation in individual populations (such as JH, MH, XM, and ML). The RIN and IRN contents were significant negatively correlated with the precipitation in May (RIRN = -0.771, p = 0.015) and June (RRIN = -0.814, p = 0.008; RIRN = -0.921, p = 0.000), indicating that precipitation was the main affecting factor of their contents. Interestingly, the analysis results showed that the RIN content had a significant positive correlation (r = 0.585, p = 0.000) with the IRN content (they are isomers); the proportion of RIN had a significant negative correlation with the sum of the two (r = -0.390, p<0.0001), while the proportion of IRN had a significant positive correlation (r = 0.390, p<0.0001). It meant that, with the total quantity of the two compounds increased, the proportion of RIN decreased and the proportion of IRN increased, illustrating that their conversion exist some regularity. Moreover, the content ratio of RIN and IRN was significant positively correlated with the January precipitation (r = 0.716, p = 0.030), implying that January may be the key period for the mutual transformation of RIN and IRN.

Polyoxypregnane Glycosides from the Roots of Marsdenia tenacissima and Their Anti-HIV Activities.

A continuous phytochemical study on the roots of Marsdenia tenacissima led to the isolation and identification of 13 new polyoxypregnane glycosides named marstenacissides B10-B17 (1, 2, 4, 7, 8, 11, 12, and 14) and marstenacissides A8-A12 (3, 9, 10, 13, and 15) in addition to two known polyoxypregnane glycosides marsdenosides M and L (5 and 6). Their structures were established by spectroscopic techniques and by comparison with the reported data in the literature. Moreover, the anti-HIV activities of these isolates and the previous isolated marstenacissides A1-A7 and B1-B9 were assessed, some of which exhibited slight or negligible effects against HIV-1.

[Endemic plants for medicine use in China].

Based on plant species databases, species lists and literature records, general situation of the Chinese medicinal endemic plant (vascular plant) has been systematically summarized, and its quantity and distribution characteristics of Chinese medicinal endemic plants are presented in this paper. The results showed that 3 150 endemic species are Chinese medicinal plants belonging to 785 genera in 153 families, which includes 38 species of 22 genera in 12 families of pteridophyta, 42 species of 14 genera in7 families of gymnosperms, and 3 070 species of 749 genara in 134 families of angiosperms. The top four families involving medicinal endemic species are Asteraceae (218 species), Ranunculaceae (182 species), Labiatae (151 species), and Liliaceae (133 species). The top four provincial administration distributed medicinal endemic species are Sichuan (1 568 species), Yunnan (1 533 species), Guizhou (955 species) and Hubei (930 species).On the regional scale, the most abundant one is the southwest region (2 465 species), followed by the central region (1 226 species) and the northwest region (949 species). Localization characteristics for domestication and artificial cultivation of medicinal endemic species are more prominent due to their narrower and limited distribution areas, indicating it is possible for these species acting as local potential resource for reasonable economic development.

[Investigation, collation and research of traditional Dai medicine of China].

In order to find out the composition, characteristics and traditional utilization characteristics of Dai medicine and promote the rational protection, inheritance and utilization of the resources and traditional knowledge of Dai medicine in China, the resources of traditional Dai medicine have been investigated systematically and the traditional knowledge of Dai medicine have been analyzed in the article. We found out that there were altogether 1 077 kinds of traditional Dai medicine in China and among which 272 were the first time recorded in the condition of Dai folk medical uses. There were 1 053 plant medicines which belong to 169 family and 694 genus. These plant medicines mainly distributed in the southern, west southern and east southern area of Yunnan province, the southern area of Guangxi, Guangdong, Guizhou, Sichuan, Fujian province and tropical, subtropical district as Taiwan, and more than 94.49% plant medicines could be found in Yunnan province. From the point of plant life form, they were major herbaceous or shrubby plants; When it is used as medicinal part, root and rhizome of plants account for the highest proportion, the next were whole plant and leaves. From nature, flavor and channel tropism points of view, the largest proportion of Dai medicines were cool, bitter-tasted and possesses water element. In terms of treatment of disease types, most of the drugs can treat gastrointestinal diseases, next were drugs that could be used to treat upper respiratory infection, traumatological and rheumatic diseases, urinary infection, gynecological diseases, hepatopathy, puerperium fever and diseases caused by poisonous insects and beast of prey bite. The study revealed that the resources of traditional Dai medicine and traditional knowledge of application were abundant in China, but the resources of traditional Dai medicine and traditional knowledge of application were faced with the risk of gradually reduce and loss. The article suggested that we should take measures to strengthen the study of protection and utilization of important traditional Dai medicine and endangered resources along with the protection and transmission of traditional knowledge of Dai nationalistic medicine.

[Effects of icariin on proliferation of vascular smooth muscle cell induced by ox-LDL via impacting MAPK signaling pathway].

This paper was aimed to study the effects of icariin (ICA) on the proliferation of vascular smooth muscle cell (VSMC) induced by oxidized low density lipoprotein (ox-LDL), and the molecular mechanism of the expression of proliferating cell nuclear antigen (PCNA) and MAPK signaling pathway. In this study, VSMC was induced by ox-LDL (50 mg•L⁻¹),the effect of ICA on the proliferation of VSMC was detected by MTT assay, Western blot and Real-time PCR. The results showed that after stimulation of ox-LDL, the proliferation activity of VSMC was increased, S phase, G2/M phase cells were increased, G0/G1 phase cells were decreased, PCNA protein expression was enhanced; ICA (40, 20, 10 µmol•L⁻¹) could effectively inhibit ox-LDL-induced VSMC proliferation, S phase and G2/M phase cells were decreased, the percentage of cells in G0/G1 phase were increased, PCNA expression was decreased, p38MAPK and ERK1/2 activation were inhibited. These results indicate that ICA can inhibit the proliferation of VSMC by reducing the expression of PCNA and blocking the p38MAPK and ERK1/2 signaling pathway.

[Quality of commercial Amomum villosum].

In recent years, with the price rise of Amomum villosum, the quality of A. villosum in the market has been in disorder. To understand the quality status of A. villosum in the market and provide reference for the commercial size fifty-seven samples were collected from different producing areas or markets from August 2013. The samples were detected with evaluation on appearance quality, determination of the contents of bornyl acetate, determination of pesticide residues and heavy metals residues based on Chinese Pharmacopoeia 2015. The results showed that the pesticide residues and heavy metals residues met the requirments, all the samples from different producing areas were qualified except one sample from Fujian province. The qualified rate of native products and imports products samples from market were 43.75% and 14.29%, respectively, the qualified rate of the samples of Yunnan province from producing areas was higher than that from the market. There are two ports at the national level in Yunnan province, where the southern herbs from. A. villosumis one of import medicines from Southeast Asia, and lots of A. villosum samples import to China from Yunnan ports. Most of pharmacists believed that all of the samples from Yunnan province produced in Yunnan. The great majority of commercial species was A. villosum, but A. longiliglare was scarce. Through the survey, it isfound that the main factors affecting the quality of Amomi Fructus was source, lots of A. villosum samples have been replaced by the Amomi Fructus, so the source of imports Amomi Fructus was not clear, which was also more difficult to identify. The quality of A. villosum needs to protect, optimize germplasm, strict control of medicinal sources, specification for medicinal harvesting and processing technology.

[Analysis phylogenetic relationship of Gynostemma (Cucurbitaceae)].

The sequences of ITS, matK, rbcL and psbA-trnH of 9 Gynostemma species or variety including 38 samples were compared and analyzed by molecular phylogeny method. Hemsleya macrosperma was designated as outgroup. The MP and NJ phylogenetic tree of Gynostemma was built based on ITS sequence, the results of PAUP phylogenetic analysis showed the following results: (1) The eight individuals of G. pentaphyllum var. pentaphyllum were not supported as monophyletic in the strict consensus trees and NJ trees. (2) It is suspected whether G. longipes and G. laxum should be classified as the independent species. (3)The classification of subgenus units of Gynostemma plants is supported.

[Research status, development and utilization of traditional Dai medicine in China].

OBJECTIVE: To provide a reference for the development and utilization of Dai medicine by investigate the present situation and existing problems of traditional Dai medicine. METHOD: Combined with the previous relevant investigations and literature in the field, the key and the development direction of traditional Dai medicine were analyzed. RESULT: The textual research, history, species, distribution, endangered resources, protection status etc. were elaborated and the key strategy of further investigation was expounded. CONCLUSION: Dai medicine resources should strengthen the basic research, such as the protection of traditional knowledge, the textual research, quality standard, chemical composition, biological activity, exploration of medicinal resources, especially monographic study on protection of major endangered medicinal resources should be intensified, which will be rise the level of development and utilization of Dai medicine resources.

[Comparison between content of saponins of Paris polyphylla var. yunnanensis in different areas of Yunnan province].

Six kinds of saponins (I, II, VII, PA, H) content of 22 samples of Paris polyphylla var. yunnanensis collected from different regions of Yunnan province were determined by HPLC, data was analyzed by SPSS 17. The results showed that the effect of altitude on saponin content was not significant, and the effect of growth area of saponins in P. polyphylla var. yunnanensis was significant, saponin content in sample from west Yunnan was significantly higher than that of samples from other regions.

[Original plant identification of Dai nationality herb "Daibaijie"].

OBJECTIVE: To identify the original plant of "Daibaijie", commonly used Dai herb. METHOD: The literature review, morphology and anatomy, pharmacognosy, molecular biology, chemistry were used to analysis. RESULT: Daibaijie's historical scientific name, Dregea sinensis Hemsl., was mistakenly given "Daibaijie" and D. sinensis have significant differences from the distribution, morphology and anatomy, pharmacognosy, molecular biology and chemical composition. "Daibaijie" matches with the characteristics of Marsdenia tenacissima (Roxb.) Moon in Flora of China in English. CONCLUSION: Daibaijie's original plant is M. tenacissima (Roxb.) Moon. The description and illustration of M. tenacissima (Roxb.) Moon in Flora of China in China are wrong. The illustration of M. tenacissima in Flora of China in English is wrong too.

[Key content and formulation of national Chinese materia medica resources survey at county level].

According to National Census for Water, National Population Census, National Land and Resources Survey, and work experience of experimental measures for national Chinese materia medica resources(CMMR) survey,the national CMMR survey at the county level is the key point of whole survey, that includes organization and management, field survey, sorting data three key links. Organization and management works of national CMMR survey needs to finish four key contents, there are definite goals and tasks, practicable crew, preparation directory, and security assurance. Field survey works of the national CMMR survey needs to finish five key contents, there are preparation works for field survey, the choice of the key survey area (samples), fill in the questionnaire, video data collection, specimen and other physical collection. Sorting data works of the national CMMR survey needs to finish tree key contents, there are data, specimen and census results.