Herpetin (HPT) is an active monomer constituent isolated from lignanoid in seeds of Herpetospermum caudigerum. HPT shows inhibitory effects in hepatic injury and HBV-DNA and the replication. In the study, we successfully prepare herpetin liposomes by film dispersion method for the first time. The prescription process was optimized, with the entrapment efficiency as the index. According to the optimized prescription, the mass ratio of HPT: phospholipids: cholesterol was 2.44:78.05: 19.51, the hydration and de-molding process was performed with 0.5% F68 solution at 50 degrees C, and the water-bath ultrasonic time was 20 min. The HPT liposomes prepared by this method showed an average entrapment efficiency of (94.50 +/- 2.15)% and a particle size of (119.2 +/- 10.7) nm, which was consistent with the trial expectations and will lay a solid foundation for the hepatic targeting delivery system in future.
Chemistry, Pharmaceutical ; methods ; Cholesterol ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Lignans ; chemistry ; isolation & purification ; Liposomes ; chemistry ; Phospholipids ; chemistry ; Ultrasonics

High-performance liquid chromatographic coupled with variable wavelength detection (HPLC-VWD) has been developed for simultaneous determination of 5 analytes including ellagic acid, quercetin, kaempferol-3-O-beta-D-rutinoside, tiliroside and kaempferol, and high-performance liquid chromatographic with an evaporative light scattering detector (HPLC-ELSD) has been established to determine goshonoside-F5 in extract of Rubi Fructus. Chromatographic separations were carried out on an Agilent ZORBAX SB-C18 column (4.6 mm x 250 mm, 5.0 microm). All calibration curves of reference standards revealed good linearity (R2 > 0.999 5) within the concentration ranges tested. The method limits of detection ranged 0.297-90.144 ng and the method limits ofquantitation ranged 0.990-300.480 ng, respectively. Recoveries of 6 analytes were from 97.11% to 101.7%, with RSD less than 2.1%. The result shows that amounts of the 6 analytes in the samples from 16 localities were found to be different. The higher latitude of growing environment, the more ellagic acid in herb. The content of total flavonoids in sample from east localities were higher than that in middle and west localities, and the content of goshonoside-F5 in Bozhou, Anhui province was higher than others. This method was found to be simple, accurate, sensitive with good repeatability. Those results might serve as a sound foundation for further study, quality control and application of Rubi Fructus.
Chromatography, High Pressure Liquid ; Ellagic Acid ; analysis ; Flavonoids ; analysis ; Geography ; Rosaceae ; chemistry

OBJECTIVETo investigate the characteristics of pain in temporomandibular disorders (TMD) through analyzing the description of pain by the TMD patients.

METHODSNinety TMD pain patients were included and the glossary in description of the intensity, rhythm and degree of disability due to TMD pain were recorded.

RESULTSThe descriptive pain by 90 patients was slight to moderate. There was no significant difference between males and females or between chronic and acute patients in the description of pain intensity (P > 0.05). The chewing disability was the most often reported complaining, and then was mental status. The probability of pain at rest was not different between in chronic pain patients and acute pain patients(P > 0.05). The acute pain patients often used aching, slight and tingle to describe their pain, while the chronic pain patients used dull, gas and numb more.

CONCLUSIONPain intensity in acute or chronic TMD patients is both low. The pain mainly influences chewing function and mental status in patients. Descriptive characteristic with chronic orofacial pain is different from acute.

Adult ; Facial Pain ; Female ; Humans ; Male ; Mastication ; Pain ; Temporomandibular Joint Disorders

OBJECTIVETo predict part of medicinal properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces by using the traditional Chinese medicinal property data prediction platform, in order to establish the relationship between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces.

METHODThe properties of traditional Chinese medicine components were predicted by using the medicinal property data prediction platform based on the pharmacological effects of the components.

RESULTThe total sum of identical or similar results of the prediction for the properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces accounted for over 75%.

CONCLUSIONThe self-similarity exists between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces, which reflects the inheritance, additivity and emergence among different properties of traditional Chinese medicines.

Drug Combinations ; Drug Prescriptions ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional

Loop-mediated isothermal amplification (LAMP) assay is a novel method of gene amplification with high specificity, sensitivity and rapidity, which can be applied for disease diagnosis in shrimp aquaculture. The method is performed under isothermal conditions with a set of four specially designed primers that recognize six distinct sequences of the target. In the present study, according to the conservative regions of non-structural protein gene NS1, a set of four specific primers were designed, and a rapid detection of IHHNV was established by LAMP assay. The parameters of reaction time and temperature were optimized, and its specificity and sensitivity were assessed. The reactions were carried out at 60 degrees C, 62 degrees C, 63 degrees C, 64 degrees C, 65 degrees C, 66 degrees C, 67 degrees C, 68 degrees C for different time (0 min; 15 min; 30 min; 45 min; 60 min; 75 min). A plasmid pMDIHHNV carrying target sequence of LAMP detection was constructed. Ten-fold serially diluted pMDIHHNV (10(7)-10(0)copies/microL) was used as template for LAMP assay to investigate the detection limit. To determine the specificity, LAMP assays were carried out with DNA templates from other pathogens (White spot syndrome virus; WSSV, Taura Syndrome Virus; TSV, Aeromonas. hydrophila, V. alginolyticus, Vibrio. parahaemolytious, Escherichia. coli). The results showed the optimized LAMP assay for the rapid detection of IHHNV was performed at 65 degrees C for 60 min. The LAMP assay had an unequivocal detection limit of 100 copies/microL, and it was 1,000 times lower than that of PCR. The nucleic acids of other pathogens were not amplified by this LAMP system with the specific primers, which showed a good specificity. The resulting amplicons were detected using visual observation after the addition of SYBR Green I and gel electrophoresis. We investigated the efficacy of UNG (uracil-N-glycosylase) and dUTP in avoiding carry-over contamination in the LAMP assay procedure and explored its effect on the amplification efficiency. Products of LAMP with dUTP adding could be lysed by UNG to avoid LAMP products carry-over contamination effectively. The LAMP assay could be finished within an hour, requiring only a regular laboratory water bath or heat block for reaction and the result could easily be detected using visual observation. Clinically suspected IHHNV-infected shrimp samples were detected by both LAMP and PCR assay, and the result indicated that IHHNV was detected rapidly by LAMP instead of by PCR.
Animals ; DNA Primers ; genetics ; Densovirinae ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Penaeidae ; virology ; Viral Proteins ; genetics

OBJECTIVETo study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).

METHODSForty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.

RESULTSThe levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.

CONCLUSIONSThe levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.

Animals ; Asthma ; drug therapy ; immunology ; Cytokines ; biosynthesis ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; Medicine, Chinese Traditional ; Phagocytosis ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th17 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVE: To explore the effect of Naoxintong capsules on learning and dysmnesia and calcitonin gene-related peptide (CGRP) about vascular dementia (VD) in rats. METHODS: The model of vascular dementia rats were built by middle cerebral artery occlusion (MCAO), and then the rats were randomly divided into a Naoxintong group (that was traditional Chinese medicine group,TCM group), a Western medicine group, a model group, a sham operation group, and a normal group. After corresponding treatment, the rat so behaviors were detected by Morris water maze experiment to obtain their performance record of learning and memory. The changes of cell morphology were detected by Nissl stain. The expression of CGRP in the hippocampus was measured with immunohistochemical staining. RESULTS: The masculine cells about CGRP of the TCM group were more than those of the model one (P<0.05). Naoxintong capsules improved the ability of learning and memory in vascular dementia rats, prevented the pyramidal cells on CA1 field in hippocampus from decreasing, increased the number of masculine cells about CGRP, and deceased the gray scale value. CONCLUSION Naoxintong capsules can raise the number of masculine nerve cells of CGRP, protect the pyramidal cells of VD rats, and lessen their learning and dysmnesia, which may be part of the mechanism of improving learning and memory capacity of VD model in rats.
Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Dementia, Vascular ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Hippocampus ; cytology ; metabolism ; Male ; Maze Learning ; Pyramidal Cells ; metabolism ; Rats ; Rats, Sprague-Dawley

Adult ; Female ; Foot Deformities, Congenital ; diagnosis ; genetics ; Hand Deformities, Congenital ; diagnosis ; genetics ; Humans ; Male ; Pedigree ; Young Adult

OBJECTIVETo evaluate the effect of Zhuanggu Jianxi Decoction (, ZGJXD) on interleukin-1 β (IL-1 β)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis.

METHODSSerum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum (ZGJXD-S) was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL-1 β stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type II collagen immunocytochemistry. After IL-1 β-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 (a specific inhibitor of p38 MAPK) for 48 h, caveolin-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL-1 β, tumor necrosis factor α (TNF-α), matrix metalloproteinase 3 (MMP-3) and MMP-13 were examined by real-time polymerase chain reaction.

RESULTSIL-1 β stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 β, TNF-α, MMP-3 and MMP-13. However, the IL-1 β-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs.

CONCLUSIONZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.

Animals ; Base Sequence ; Blotting, Western ; Caveolins ; metabolism ; Chondrocytes ; drug effects ; enzymology ; metabolism ; DNA Primers ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; Interleukin-1beta ; physiology ; MAP Kinase Signaling System ; Male ; RNA, Messenger ; genetics ; Rabbits ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo understand the biochemical characteristics, virulence genes and pathogenicity of Shigella flexneri Xv isolated in Beijing.

METHODS61 strains of S. flexneri Xv isolated from diarrhea patients in Beijing were systematically determined through biochemical reactions and serological tests. Application of PCR technique in detection of virulence genes on ipaH, sen, virF, ial and pulsed-field gel electrophoresis (PFGE) was used to identify the related characteristics and on rat lung slices to determine its pathogenicity.

RESULTSAll of the S. flexneri Xv could ferment glucose, mannitol, melibiose and arabinose. Using serum agglutination, we found that the antigen structure was (IV: 7, 8). IpaH, sen, virF and ial that carried rates of virulence genes appeared to be 100%, 81.97%, 75.41% and 80.30%, respectively. Among 61 strains of S. flexneri Xv, the PFGE typing of Shigella bacteria could be divided into 25 belt types while the results from rat lung slices showed inflammatory change of Xv.

CONCLUSIONS. flexneri Xv was found that it carried high rate of Shigella virulence genes, exhibiting genetic polymorphism and highly invasive.

Animals ; Humans ; Microbial Sensitivity Tests ; Rats ; Shigella flexneri ; classification ; isolation & purification ; pathogenicity ; Virulence ; genetics