High resolution melting (HRM), an important technology for genotyping and mutation scanning, has broad prospects in the authentification of traditional Chinese medicine. This paper selected universal trnH-psbA primers and used HRM to establish a new methods for identification of Akebia herbs. PCR was conduct at the annealing temperature of 58 degrees C and 35 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further analyzed. The results showed the Tm values of Caulis Akebiae was (81.84 ± 0.16), (85.28 ± 0.16) degrees C and Caulis Clematidis Armandii was (83.22 ± 0.19) degrees C and Caulis Aristolochiae manshuriensis was (81.67 ± 0.14) degrees C, (84.24 ± 0.10) degrees C with 5-125 mg - L-' DNA template, 0.4 μmol x L(-1) primer, 2.0 mmol x L(-1) Mg2+. This method can achieve the authentification of Akebia herbs and is simple, fast, high-throughput, visual.
Chemistry Techniques, Analytical ; methods ; DNA, Plant ; chemistry ; genetics ; Genotype ; Magnoliopsida ; chemistry ; classification ; genetics ; Phylogeny ; Transition Temperature

OBJECTIVETo study the in-vitro inducing apoptosis mechanism of human hepatoma SMMC-7721 cells by 2',4'-di- hydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC), a chalcone compound from Cleistocalyx operculatus.

METHODQuantitative DNA fragmentation assay was carried out to detect the effect of DMC of different concentrations on SMMC-7721 cells, according to the method of Sellins and Cohen with some modifications. Telomerase activities of the cells were determined by PCR-ELISA methods. The expression quantity of c-myc and hTERT mRNA were determined by semi-quantitative RT-PCR The effect of DMC on expression levels of cmyc and hTERT protein were measured by western blot.

RESULTThe percentage of DNA fragmentation increased with notable concen- tration dependence, after treatment with DMC for 48 h. Compared with that of control group, the telomerase activity of the cells de- creased by (66.2 ± 2.1)% after 48 h treatment with 20 μmol x L(-1) DMC, the mRNA expression of c-myc and hTERT decreased by (67.3 ± 2.1)% and (64.4 ± 2.3)%, respectively, and the protein expression of c-myc and hTERT decreased by (69.6 ± 1.9)% and (71.3 ± 2.4)%, respectively.

CONCLUSIONDMC can induce SMMC-7721 cell apoptosis and the apoptosis mechanism may be related to the decreased mRNA and protein expression of c-myc and hTERT.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Chalcones ; pharmacology ; DNA Fragmentation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Liver Neoplasms ; pathology ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Syzygium ; chemistry ; Telomerase ; genetics ; metabolism

BACKGROUND:With the development of modern pathological techniques, the misdiagnosis rate has been reduced remarkably, but special stains are still the most important method for pathological diagnosis. OBJECTIVE:To compare the advantages and disadvantages of different special stains used for observing the structure of human lumbar facet joints. METHODS:The specimens of facet joint cartilage at L4/5 level were collected from patients undergoing lumbar surgery, and then stained with hematoxylin-eosin, safranin O, toluidine blue, Masson, and saranin-O-fast green for structure observation. RESULTS AND CONCLUSION:The structure of the articular cartilage could be observed clearly through hematoxylin-eosin, toluidine blue, and saranin-O-fast green staining. The cartilage surface, tidemark, and subchondral bone were shown by the hematoxylin-eosin staining, with the presence of violet chondrocyte nuclei. Safranin-O-fast green staining showed the four layers of the cartilage clearly, including the shallow layer (cartilage surface), middle layer (spherical cells arranged in disorder), columnar cell layer (large and multinucleated chondrocytes arranged neatly), tidemark, subchondral bone layer; and the cartilage matrix was reddish uniformly, the subchondral bone was green, and the cartilage and bone tissue showed a striking contrast. The cartilage structure was unclear in toluidine blue staining, with clear nuclei and almost no coloring cytoplasm, but the matrix appeared with slight purplish blue. Safranin O staining showed that the cartilage was red, which had no obvious boundary with the cartilage matrix, and chondrocytes were stained lightly. Masson staining showed clear collagen fibers, but the structures of the cartilage and subchondral were obscure. To conclude, safranin-O-fast green staining can achieve the best results, followed by hematoxylin-eosin staining and Masson staining in turn.

Objective To investigate the relationship between the genetic polymorphisms, which played roles in the pathogenesis of metabolic syndrome (MS), and susceptibility of non-alcoholic fatty liver disease (NAFLD) in Han people in Guangdong province. Methods The subjects were selected from an epidemiologie survey in Guangdong province. Fifty to 117 adult NAFLD patients, who met the criteria of Chinese guideline for diagnosis of NAFLD and had typically clinical, biochemical signs and abdominal ultrasonography, were recruited in the study. By using 1 : 1 matched method of nested case-control study, same numbers of people without NAFLD were included as controls. The genetic analyses was performed by using genomic DNA extracted from peripheral blood. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) at 9 sites in 7 candidate genes. Results Most SNPs of the genes were related to the susceptibility of NAFLD. Some of them had positive relation (increasing the risk) such as tumor necrosis factor (TNF)-α-238, adiponectin-45, leptin-2548, peroxisome proliferator-activated receptors (PPAR) γ-161 and phosphatidylethanolamine N-methyltransferase (PEMT)-175. Some had negative relation (decreasing the risk) including adiponectin-276 and hepatic lipase-514. And some had no relation (TNF-α-380 and PPAR g coactivator-1α-482). Conclusion Most cytokines' SNPs of candidate genes discovered in MS patients are related to the susceptibility of NAFLD.

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

To develop a reverse dot blot assay for rapid detection of HBV genotypes.Specific oligonucleotides probes were desighed and immobilized on nylon membranes.The DNA sample to be tested was PCR-amplified with DIG labeling primers and then hybridized with the immobilized probes.This procedure for detecting HBV genotypes was simple,rapid and specificity.30 specimens in Chongqing area were collected and detected by this method,and results were evaluated using direct sequencing.Results showed that: This new method was applicable to precise detection HBV genotypes for specimen with copies up to 103,and the HBV genotyping results showed that genotype B was the predominant genotype in Chongqing area.

OBJECTIVEIt laid the foundation of the large sample data mining for a comprehensive summary concerning four properties theory of traditional Chinese medicine (TCM), and also provided theory clues on determination of four properties for the new resource development of TCM and the clinical use of Chinese medicine.

METHODFour properties data of 8 980 Chinese medicines from "Chinese herbal medicine (CHM)" and associated function index data were chose as data sets. Then, the IBM SPSS Clementine 14.1 data mining platform and Apriori model were adopted to mining classification-association rules, setting the minimum support threshold of rule antecedent and the minimum confidence threshold as 0.5% and 60%.

RESULT11 classification-association rules involved in warm, cold and mild natures were found.

CONCLUSIONIt was discovered that the TCM with functions of "dispelling wind-cold, warming the middle, stopping pain and expelling wind-removing dampness, tonifying kidney yang, unblocking meridians and expelling wind-removing dampness, dispelling cold to stop pain, strengthening sinews-bones and expelling wind-removing dampness" was likely warm-natured, with functions of "tonifying the lung" was likely mild-natured, and with functions of " clearing heat and drying dampness, clearing heat and purging fire, eliminating restlessness" was likely cold-natured.

Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Phytotherapy

OBJECTIVEIn order to provide theoretical clues and data support for the use of Chinese medicine clinical drug, experimental study of Chinese materia medica and development of new resources of Chinese materia medica, the four properties as the core, the relationships of property, flavor, channel tropism and toxicity in nature data of Chinese materia medica were analyzed.

METHODThe spearman rank correlation method was employed to analyze 8 356 Chinese drugs with characteristic of four properties from " Chinese Herbal Medicine" based on data level coding.

RESULTIt was discovered that four properties showed significant positive correlations with tastes of "pungent and sweet" , channels of "spleen" , "stomach" , "kidney" and "toxicity" , but also showed significant negative correlations with tastes of "bitter" and "light" and six channels such as "large intestine" , "heart", "bladder" , "gallbladder" , "small intestine" and "lung" (in descending order of correlation ) (P <0. 01).

CONCLUSIONIt was indicated that the more hot the Chinese medicine nature, the more possible it contained "toxicity" , tastes of "pungent" and "sweet" , and the more possible it was belong to channels of "spleen" , "stomach" and "kidney". As well, the more cold the Chinese medicine nature, the more possible it contained tastes of "bitter" and "light", and the more possible it was belong to six channels such as "large intestine", etc.

Herbal Medicine ; methods ; Materia Medica ; standards ; Medicine, Chinese Traditional ; methods ; standards

OBJECTIVETo observe the histological and ultrastructural changes of the skin and hair follicles following hair removal by alexandrite laser in Tibet mini-pigs.

METHODSTwelve healthy Tibet mini-pigs with dark hair were treated with alexandrite laser for hair removal. The skin specimens were taken immediately and at 1 h and 1, 3, 5, 10, 15, 30, 60 days after the laser treatment for observation under optical and transmission electron microscope.

RESULTSLaser hair removal resulted in extensive coagulation necrosis, carbonization and falling of the subcutaneous hair shafts, and some of the cells in the outer root sheath and hair bulb underwent degenerative and necrotic changes. One hour after laser treatment, the cells in the outer root sheath and bulb exhibited nuclear condensation, fragmentation and or karyolysis characteristic of cell apoptosis. The cell apoptosis reached the peak level on day 3 after the laser exposure, accompanied by endothelial degeneration in the hair papilla vessels, edema and lymphocyte infiltration in the dermal tissues. Tissue reaction and inflammation were relieved on day 5, and the dermal tissue and follicles recovered their normal structures on day 10. At 60 days after the treatment, the hair follicles decreased markedly but the structure of the residue follicles remained normal.

CONCLUSIONAlexandrite laser exposure results in selective destruction of the follicles by inducing direct coagulation and cell apoptosis to achieve permanent hair removal. Tibet mini-pigs with black hair can be used as the animal model of clinical laser hair removal.

Animals ; Hair Follicle ; radiation effects ; ultrastructure ; Hair Removal ; methods ; Lasers, Solid-State ; therapeutic use ; Microscopy, Electron, Transmission ; Swine ; Tibet

BACKGROUND: Adiponectin is one of the adipocytokines that have been found to be negatively correlated with obesity in recent years. As an endogenous bioactive peptide, adiponectin is shown to be associated with insulin resistance, type 2 diabetes mellitus, metabolic syndrome, and participate in the regulation of bone metabolism. OBJECTIVE: To describe the mechanism of adiponectin in bone metabolism, and mainly clarify the correlation of adiponectin with osteoblasts and osteoclasts. METHODS: The first author searched PubMed and CNKI databases for the articles addressing the experimental research and research progress of adiponectin in bone metabolism published from 2003 to 2017. The keywords were "adiponectin, osteoporosis, bone metabolism, osteogensis, bone absorption" in English and Chinese, respectively. Adiponectin, its receptors, bone metabolism, adiponectin effects on bone formation and bone resorption were reviewed. RESULTS AND CONCLUSION: Adiponectin exerts a comprehensive effect on bone metabolism, mainly through autocrine, paracrine and endocrine, insulin signaling pathways. Besides, it exerts biological effects mostly by regulating OPG/RANKL/RANK system, especially in bone formation and bone resorption. Therefore, adiponectin may prevent osteoporosis by regulating bone metabolism.