[Multiplex PCR method for simultaneous identification of Xanthii Fructus and its adulterants].

The purpose of this study is to establish a molecular method to identify Xanthii Fructus and two adulterants, the fruits of Xanthium mongolicum and X. italicum. Xanthii Fructus is the fruit of X. sibiricum, which is a Chinese herbal medicine used clinically to treat allergic rhinitis. The fruits of X. mongolicum and X. italicum have strong morphological similarities with Xanthii Fructus, while their safety of medication cannot be guaranteed. The genomes of X. sibiricum, X. mongolicum, and X. italicum were sequenced, which generated sequences of 2.21, 2.24, and 2.54 Gb, respectively. Based on the 76 specific contigs screened out by BLASTN and Bowtie 2, the corresponding primers were designed by Primer 5.0. Three pairs of primers with stable amplification efficiency and good reproducibility were screened out to establish a multiplex PCR method based on the PCR amplification results. Further, the annealing temperature, the amount of DNA template, the number of cycles, different DNA polymerases, and different PCR thermal cyclers were optimized. Fragments of 262 bp and 458 bp from X. sibiricum, 260, 454, and 927 bp from X. mongolicum, and 260 bp and 926 bp from X. italicum were amplified under the following conditions: the annealing temperature of 52 ℃, 35 cycles, 30 ng template DNA. Then, the established method was used to detect 18 samples of X. sibiricum, 17 samples of X. mongolicum, and 12 samples of X. italicum. The results showed that all the samples had positive results, which were consistent with the morphological identification results, thus proving the stability and reliability of the established method. Combining genome sequencing technology and multiplex PCR method to identify Xanthii Fructus and its adulterants can not only obtain the difference in genetic background but also facilitate the design of reliable primers. The multiplex PCR have high specificity and repeatability, providing a new method for the molecular identification of Xanthii Fructus.

[Study on geographical variation of medicinal ingredients contents in Scutellaria baicalensis from different provenances].

In order to investigate the genetic difference on medicinal components of Scutellaria baicalensis from different provenances on the genetic difference, the S. baicalensis provenance tests were arranged by randomized block design.Excavating the crude drugs that have been growing for three years, with the same drying process, the content of baicalin, baicalein, wogonoside, wogonin and laminarin A in S. baicalensis were detected by HPLC, and then the data were analyzed. The results indicated that the content of baicalin in different provenances of S. baicalensis was significantly different (P<0.05), while the variation of baicalein reached extremely significant level (P<0.01). Cluster analysis showed that if the distance was divided by 5.0, the provenances in Chengde, Hebei province were divided into two independent populations, while the other two populations had large geographic spans. The results show that the significant geographical variations exist in the content of medicinal components in S. baicalensis. The study laid a theoretical foundation of provenance selection of S. baicalensis.

[Genotype, environment and their interactions of functional components in Rheum palmatum].

Eight kinds of provenance of Rheum palmatum collected from 4 provinces Sichuan, Ningxia, Gansu, Shannxi as test materials, which were transplanted under 3 different environments by using complete randomized block design with three replicates. The contents of the chemical components was determined by HPLC. This study aimed at analyzing the effect of genotype, environment and their interactions on the 4 kinds of functional components (phenolic acids, bianthrone, free anthraquinones and combined anthraquinones) in 14 kinds of active components of R. palmatum, in order to provide a theoretical basis for the selection of cultivated R. palmatum in high quality producing area and excellent provenance. The functional components of R. palmatum were influenced by genotype and environment. The content of phenolic acids was mainly influenced by environment, and the other three kinds of functional components were affected by environment and their interactions. The proportion of environment was larger. The cultivation quality of R. palmatum should give priority to environment, then choose a provenance. Sichuan may be beneficial in accumulation of free anthraquinones in R. palmatum, Gansu may facilitate the binding of combined anthraquinone, phenolic acids and bianthrone content. Preliminary inference based on the content and proportion of efficacy components, P2 could be potential special medicinal germplasm that have function of heat-clearing and detoxifying drugs. P6 could be potential special medicinal germplasm that activate blood circulation to dissipate blood stasis. P7 and P1 could all be potential specialmedicinal germplasms that exist diarrhea attack characters. The results of this study have important guiding significance for the production of rhubarb precision medicinal materials.

[Genotype, environment and their interactions of functional components in Rheum tanguticum].

Six kinds of provenance of Rheum tanguticum collected from Qinghai province as the test materials, which were transplanted under 3 different environments by using complete randomized block design with three replicates. The contents of the chemical components was determined by HPLC. This study aimed at analyzing the effect of genotype, environment and their interactions on the 4 kinds of functional components (phenolic acids, bianthrone, free anthraquinones and combined anthraquinones) in 14 kinds of active components of Rh. tanguticum, in order to provide a theoretical basis for the selection of cultivated Rh. tanguticum in high quality producing area and excellent provenance. The provenance trial showed that the genotype and environment influence on the effect of all kinds of functional components in Rh. tanguticum were significant (P<0.05). The content of phenolic acids was mainly influenced by environment, and the other three kinds of functional components were affected by environment and their interactions. The proportion of environment was larger. The cultivation quality of Rh. tanguticum should give priority to environment, then choose a provenance. Sichuan may be beneficial in accumulation of combined anthraquinones in Rh. tanguticum, Gansu may facilitate the binding of free anthraquinone, phenolic acids and bianthrone content. Preliminary inference based on the content and proportion of efficacy components, T4 could be potential special medicinal germplasm that have function of heat-clearing and detoxifying drugs and activate blood circulation to dissipate blood stasis; T3 and T6 could all be potential specialmedicinal germplasms that exist diarrhea attack characters. The results of this study have certain guiding significance for the production of rhubarb precision medicinal materials.

[Influence of cutting seedling on growth, quality and yield of both aerial and underground part by cutting seedling in Scutellaria baicalensis].

By measuring the growth data of Scutellaria baicalensis in different cutting-seedling and determined active ingredient contents by HPLC and ultraviolet spectrophotometric determination. such as flavonoids. baicalin. wogonoside. baicalein. wogonin. oroxylin A. scutellarin. luteolin. and apigenin in the whole plant. Under circumstances of guaranteeing the quality and yield of medicinal materials. the yield of medicinal materials. and stems and leaves reached 193.60,63.21 kg/mu after twice cutting seedling. Not only yield but also active ingredient contents have been improved to some extent. the contents of flavonoids. baicalin. wogonoside. baicalein. wogonin. oroxylin A reached 18.52%. 15.13%. 4.03%. 1.04%. 1.04%. 0.12%. respectively in roots. Luteolin was not detected in young stems and leaves of S. baicalensis,the contents of other active ingredients such as scutellarin. luteolin and apigenin reached 7.00%. 0.96%. 0.04% respectively under twice cutting seedling. Therefore. regular cutting seedling could be regard as a new cultivation technique for wider range of promotion. And gaining high quality and yield of medicinal materials and tea with the purpose of rational utilization of natural resources and promoting the development of integration of herbal combination.

[Acquisition and storage methods for image data collected in Chinese medicine resources survey].

The acquisition and storage of the image data are important in the Chinese medicine resources survey, and it is important data and evidence for the process and the results. The image data of the Chinese medicinal materials' habitat, original plant or animal, processing in habitat, commodity form, the relative contents and workshop scenarios in the investigation are important for the compiling of the Color Atlas of National Chinese Medicine Resources, mapping the digital scattergram of the Chinese medicine resources, establishing the digital Chinese medicine plant herbarium and acquiring the documentary of the Chinese medicine resource survey. The content, procedures and methods of the video data collecting have been related and analyzed in this article to provide reference for the Chinese medicine resources survey.

Cardioprotective effect of water and ethanol extract of Salvia miltiorrhiza in an experimental model of myocardial infarction.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza has long been used in the traditional Chinese formulations for the treatment of heart ischemic diseases. AIM OF THE STUDY: We investigated the cardioprotective effect of purified Salvia miltiorrhiza extract (SME) in an experimental model of acute myocardial infarction. MATERIALS AND METHODS: Following induction of acute myocardial infarction in rats by adminstration of isoproterenol, hemodynamic and electrocardiographic parameters were monitored and recorded continuously, cardiac enzymes and parameters of oxidative stress were measured, and histopathological examination of heart tissue was performed. Experiments were performed in rats treated with SME or vehicle, as well as in those treated with Fufang Danshen Tablet (FDT) as a positive control which has previously been shown to prevent myocardial ischemia. RESULTS: Isoproterenol-treated rats showed reductions in left ventricular systolic pressure as well as in maximum and minimum rate of developed left ventricular pressure, together with an increase in left ventricular end-diastolic pressure. They also demonstrated ST-segment elevation, together with increases in serum levels of lactate dehydrogenase, glutamic oxalacetic transaminase, creatine kinase and malondialdehyde, as well as decreases in serum activities of glutathione peroxidase and superoxide dismutase. Oral administration of SME (29.76 or 59.52 mg/kg) blunted all of the hemodynamic and biochemical changes induced by isoproterenol, as did FDT (1210 mg/kg). The protective effect of SME on isoproterenol-induced myocardial damage was further confirmed by histopathological examination. CONCLUSIONS: Our results suggest that SME affords protection against isoproterenol-induced myocardial infarction.

Aqueous extract of Gleditsia sinensis Lam. fruits improves serum and liver lipid profiles and attenuates atherosclerosis in rabbits fed a high-fat diet.

ETHNOPHARMACOLOGICAL RELEVANCE: Gleditsia sinensis Lam. has been used in the traditional Chinese medicine as a chief ingredient of many polyherbal formulations for the treatment of obesity and thrombosis. AIM OF THE STUDY: To evaluate the effects of Gleditsia sinensis Lam. fruit aqueous extract (GAE) on hyperlipidemia and atherosclerosis in Japanese white rabbits on a high fat diet. MATERIALS AND METHODS: Rabbits were divided into four groups: the normal control with a normal diet, and high-fat diet-fed model group and GAE-treated groups supplemented with GAE (6 or 12 mg/kg/day, p.o.), respectively. The groups fed high-fat diets were given i.v. with bovine serum albumin (BSA) on the 4th week to induce atherosclerosis. The serum lipid profile, including triglycerides (TG), total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C), was determined on the 0th, 4th, 8th and 14th week, respectively. And the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine kinase (CK) were measured simultaneously. At the end of the experiment, the rabbits were sacrificed, and the atherosclerotic plaques as well as the histopathological changes of aorta and liver were assessed by oil-red or HE staining, respectively, and the aorta and liver lipid profiles were also assayed. RESULTS: Results showed that the prophylactic treatment with GAE could significantly decrease the lipid levels of serum, aorta and liver, attenuate aortic atherosclerosis and improve aortic remodeling without the significant liver and muscle toxicity. CONCLUSION: The present findings suggest that GAE can effectively attenuate the atherosclerotic at least through anti-hyperlipidemic activity and thus has the therapeutic potential in treating hyperlipidemia-related cardiovascular diseases.

Brain glucose metabolic changes associated with chronic spontaneous pain due to brachial plexus avulsion: a preliminary positron emission tomography study.

BACKGROUND: Previous brain imaging studies suggested that the brain activity underlying the perception of chronic pain may differ from that underlying acute pain. To investigate the brain regions involved in chronic spontaneous pain due to brachial plexus avulsion (BPA), fluorine-(18)fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) scanning was applied to determine the glucose metabolic changes in patients with pain due to BPA. METHODS: Six right-handed patients with chronic spontaneous pain due to left-BPA and twelve right-handed age- and sex-matched healthy control subjects participated in the (18)F-FDG PET study. The patients were rated by visual analog scale (VAS) during scanning and Hamilton depression scale and Hamilton anxiety scale after scanning. Statistical parametric mapping 2 (SPM2) was applied for data analysis. RESULTS: Compared with healthy subjects, the patients had significant glucose metabolism decreases in the right thalamus and SI (P < 0.001, uncorrected), and significant glucose metabolism increases in the right orbitofrontal cortex (OFC) (BA11), left rostral insula cortex and left dorsolateral prefrontal cortex (DLPFC) (BA10/46) (P < 0.001, uncorrected). CONCLUSION: These findings suggest that the brain areas involved in emotion, attention and internal modulation of pain may be related to the chronic spontaneous pain due to BPA.

[Clinical characteristics of patients with chronic fatigue syndrome: analysis of 82 cases].

OBJECTIVE: To analyze the clinical characteristics of Chinese patients suffering from chronic fatigue syndrome (CFS) and provide clinical and laboratory evidence for the study of its etiology and treatment. METHODS: 82 patients with CFS diagnosed based on the CDC criteria 1994 were recruited. History was collected, and physical examination was made. SCL-90 and memory test were used, and Hamilton Anxiety Rating Scale was used to those showing depression and/or anxiety. Laboratory examination, including examination of electrolytes, blood sugar, creatinine, bilirubin, alkaline phosphatase, alanine trasaminase, etc, was conducted. Western blotting was used to detect the protein-24 of Borna disease virus (BDV) in the plasma of 61 patients and 73 healthy controls. High-pressure chromatography was conducted to detect n-6 fatty acids on the membrane of erythrocytes of 42 patients and 37 healthy controls. Plasma L-carnitine in 61 patients and 73 healthy controls was detected by zymological analysis. In different examinations sex and age-matched controls were used. RESULTS: Most of the patients were 21 approximately 50 years old (74/82, 90.24%). No gender difference was found. The patients usually had 4 approximately 6 symptoms besides distinctive fatigue. Descent of remembrance and/or attention was the most conspicuous accompanying symptoms (69/82, 84.15%). Abnormalities in SCL-90 scores were present in 57 patients (69.51%), e.g, somatization existed most commonly (32/82, 39.02%), and anxiety and depression were 20.73% (17/82) and 18.29% (15/82) respectively. The prevalence of anti-BDV-p24 antibody was 20.73% (17/82), significantly higher than that of the controls (0%, chi(2) = 6.673, P = 0.010). The arachidonic acid level was significantly lower in the CFS group than in the controls (P > 0.05) and there were no differences in linoleic acid and ETA (both P > 0.05). The level of L-carnitine was 6.4336 +/- 3.4225, significantly lower than that of the control group (7.6666 +/- 3.5819, t = 2.025, P = 0.045) and the L-carnitine level was increased 2 weeks after supplementary treatment, together with improvement of symptoms. CONCLUSION: Most of the CFS patients are young and middle-aged. Descent of reorganization is common in these patients. Psychological abnormalities exist in most patients. Some patients are infected with BDV, some with deficiency of nutrition and/or abnormality of energy metabolism.

[Neuroelectrophysiological basis and surgical treatment of essential tremor].

OBJECTIVE: To investigate the relationship between limb tremor and neuronal firing in thalamus (Vim) and retrospectively review the clinical effects and safety of the surgical treatment of essential tremor (ET). METHODS: Forty-two ET patients received microelectrode-guided thalamotomy and 11 cases were quantitatively evaluated with FAHN rating scales pre- and post-operatively. RESULTS: There were electrophysiological tremor-related neurons in ventrolateral part of thalamus. Lesioning of those neurons abolished contralateral limb tremor in all of the patients. No permanent contralateral weakness, dysarthria and hemorrhage were observed. CONCLUSION: "Tremor cell" in thalamus plays a key role in the symptom of ET patients. Destruction of those cells may completely and permanently abolish tremor symptom.