Fermented Chinese medicine has long been used. Amid the advance for preservation of experience, the connotation of fermented Chinese medicine has been enriched and improved. However, fermented Chinese medicine prescriptions generally contain a lot of medicinals. The fermentation process is complicated and the conventional fermentation conditions fail to be strictly controlled. In addition, the judgment of the fermentation end point is highly subjective. As a result, quality of fermented Chinese medicine is of great difference among regions and unstable. At the moment, the quality standards of fermented Chinese medicine are generally outdated and different among regions, with simple quality control methods and lacking objective safe fermentation-specific evaluation indictors. It is difficult to comprehensively evaluate and control the quality of fermented medicine. These problems have aroused concern in the industry and also affected the clinical application of fermented Chinese medicine. This article summarized and analyzed the application, quality standards, and the modernization of fermentation technology and quality control methods of fermented Chinese medicine and proposed suggestions for improving the quality standards of the medicine, with a view to improving the overall quality of it.
Medicine, Chinese Traditional ; Reference Standards ; Quality Control ; Fermentation

Objective:To investigate the taxonomic structure and diversity of endophytic fungi from Datura metel and screen the strains with anti-dermatophyte activities， so as to provide resources for the development of new lead compounds against dermatomycosis. Method:Endophytic fungi were isolated from the roots， stems and leaves of D. metel after tissue block incubation and then identified by morphological analysis and rDNA-internal transcribed spacer（ITS） sequencing. Their anti-dermatophyte activities were detected by agar diffusion assay. Result:A total of 292 strains of endophytic fungi were isolated from D. metel， belonging to 34 genera， with Fusarium （72.97%） in roots， Fusarium （37.25%） and Plectosphaerella （28.43%） in stems， and Colletotrichum （39.66%） in leaves as the dominant species. The isolation rate （89.23%）， colonization rate （84.62%）， and diversity index （1.82） of endophytic fungi in leaves were significantly higher than those in roots （70.48%， 70.48% and 1.23） and stems （69.39%，68.03% and 1.64）. The determination of anti-dermatophyte activities of 35 endophytic fungal fermented filtrates showed that the strains exhibiting inhibitory activities against Microsporum canis， Trichosporon mucoides， Trichophyton rubrum and Candida albicans accounted for 97.14%， 71.43%， 45.71%， and 25.71%， respectively. Among them， six strains （17.14%）， namely Fusarium sp. R1， Penicillium sp. R5， Aspergillus sp. R7， Metarhizium sp. S18， Diaporthe sp. S19， and Glomerella sp. L57， all inhibited the four types of cutaneous fungal pathogens. Conclusion:The endophytic fungi in D. metel are diverse， and the proportion of endophytic fungi possessing anti-dermatophyte activities is high， allowing them to serve as potential resources for the development of new anti-dermatophyte agents.

Objective:To study the changes in total phenolic and flavonoid content and antioxidant enzyme activity of Polygala tenuifolia callus in MS medium with different concentrations of H₂O₂,in order to explore the physiological mechanism of Polygala tenuifolia callus in adapting to H₂O₂ environmental stress at the cellular level. Method:Five gradients of 0,5,10,15,20 mmol·L^-1 were set for H₂O₂ concentration and added to MS medium, with P. tenuifolia callus as the experimental material. Total phenols,total flavonoids and antioxidant enzyme activities of callus were determined after 5,10,15,20,25 d of culture,respectively. Result:The contents of total phenols and flavonoids were the highest when the concentration of H₂O₂ was 5 mmol·L^-1 for 15 d. The SOD activity was the highest when the callus was cultured for 5 d and the exogenous H₂O₂ concentration was 5 mmol·L^-1. POD activity was the highest at 25 d and 5 mmol·L^-1 concentration of exogenous H₂O₂.CAT activity was the highest at 25 d and 15 mmol·L^-1 concentration of exogenous H₂O₂. Conclusion:P. tenuifolia callus has the ability to adapt to the environmental stress of H₂O₂ at a certain concentration. When it is subjected to the environmental stress of H₂O₂,P. tenuifolia callus can alleviate the damage by regulating its secondary metabolites and protecting enzyme system. It can significantly promote the content of total phenols and flavonoids in secondary metabolites at 5-10 mmol·L^-1. SOD activity was significantly increased at 5 d and the concentration of exogenous H₂O₂ of 5 mmol·L^-1. POD activity was significantly increased at 25 d and the concentration of exogenous H₂O₂ of 5 mmol·L^-1. CAT activity was significantly increased at 25 d and concentration of exogenous H₂O₂ of 15 mmol·L^-1.

Objective:To study the effect of different hormone ratios on the callus induction of roots,stems and leaves of Polygala tenuifolia,and determine and analyze the amount of flavonoids in roots,stems and leaves of P. tenuifolia. Method:With MS as the basic medium and roots,stems and leaves of P. tenuifolia sterile seedings as explants,the effects of 2,4-D,NAA and 6-BA on callus induction and flavonoid accumulation in different parts of roots,stems and leaves of P. tenuifolia were determined by orthogonal test. Result:2,4-D,NAA and 6-BA had significant effects on the callus induction rate of roots,stems and leaves of P. tenuifolia. The optimal callus induction combination of leaves was MS+3.0 mg·L^-1 2,4-D+1.0 mg·L^-1 NAA+1.5 mg·L^-1 6-BA,the optimal callus induction combination of stems was MS+1.0 mg·L^-1 2,4-D+3.0 mg·L^-1 NAA+1.5 mg·L^-1 6-BA,the optimal callus induction combination for roots was MS+1.0 mg·L^-1 2,4-D+1.0 mg·L^-1 NAA+1.0 mg·L^-1 6-BA. And 2,4-D,NAA and 6-BA had significant effects on flavonoid accumulation in the stem callus of P. tenuifolia,and MS+3.0 mg·L^-1 2,4-D+1.0 mg·L^-1 NAA+0.5 mg·L^-1 6-BA was the best flavonoid accumulation combination.NAA,6-BA had significant effects on flavonoid accumulation in the leave callus of P. tenuifolia,while 2,4-D had no significant effect,and MS+3.0 mg·L^-1 2,4-D+2.0 mg·L^-1 NAA+1.0 mg·L^-1 6-BA was the optimal flavonoid accumulation combination,the three hormones had no significant effect on the accumulation of flavonoids in the root callus of P. tenuifolia,and MS+2.0 mg·L^-1 2,4-D+1.0 mg·L^-1 NAA+1.5 mg·L^-1 6-BA was the best flavonoid accumulation combination. Conclusion:Under the conditions,the callus induction rate of roots,stems and leaves of P. tenuifolia is 100%, especially, the callus of P. tenuifolia leaves was the optimal,which is followed by P. tenuifolia stems and P. tenuifolia roots. Under the conditions,the amount of flavonoids in roots,stems and leaves of P. tenuifolia reach 21.31,24.56,23.61 mg·g^-1,respectively.

OBJECTIVE: To establish a novel method to isolate endothelial progenitor cells（EPC） from cryopreserved umbilical cord blood (cryoUCB), to investigate the biological characteristics of EPC and to improve the rate of EPC obtained from cryoUCB. METHODS: Twelve cryoUCB samples during 2000 to 2001 years were collected from allogeneic cord blood bank, cryoUCB was thawed rapidly in a water bath at 37 ℃, total nucleated cells (TNCs) were washed by phosphate-buffered saline (PBS). TNCs were seeded onto fibronectin-coated dishes to isolate EPC. Flow cytometry and immunofluorescence were used to identify EPC. The function of EPC was identified in vitro, such as the incorporation of Dil-Ac-LDL and FITC-UEA-I, the formation of capillary-like structure in matrigel, and the release of VEGF by ELISA. RESULTS: One to five cluster of cobble stone-like cells appeared at 2-3 weeks after seeding. Flow cytometric analysis showed that positive rates of CD31, CD34, CD144, and VEGFR (CD309) were（92.91±5.20）%, （30.0±23.27）%, （88.55±3.83）% and （67.21±12.12）% in passage 1 to passage 3 of EPC. EPC could uptake Dil-Ac-LDL and FITC-UEA-I, form capillary-like network on Matriget and release VEGF. CONCLUSION EPC had been successfully isolated from cryopreserved umbilical cord blood by this method with high stability and reproducibility. EPC can be obtained in 85% frozen umbilical cord blood. This method may lay a foundation to supply abundant EPC for clinical application.
Cell Differentiation ; Cells, Cultured ; Endothelial Progenitor Cells ; Fetal Blood ; Reproducibility of Results ; Stem Cells

BACKGROUND/OBJECTIVES: Docosahexaenoic acid (DHA), an n-3 long chain polyunsaturated fatty acid (LCPUFA), is acquired by dietary intake or the in vivo conversion of α-linolenic acid. Many enzymes participating in LCPUFA synthesis are regulated by peroxisome proliferator-activated receptor alpha (PPARα). Therefore, it was hypothesized that the tissue accretion of endogenously synthesized DHA could be modified by PPARα. MATERIALS/METHODS: The tissue DHA concentrations and mRNA levels of genes participating in DHA biosynthesis were compared among PPARα homozygous (KO), heterozygous (HZ), and wild type (WT) mice (Exp I), and between WT mice treated with clofibrate (PPARα agonist) or those not treated (Exp II). In ExpII, the expression levels of the proteins associated with DHA function in the brain cortex and retina were also measured. An n3-PUFA depleted/replenished regimen was applied to mitigate the confounding effects of maternal DHA. RESULTS: PPARα ablation reduced the hepatic Acox, Fads1, and Fads2 mRNA levels, as well as the DHA concentration in the liver, but not in the brain cortex. In contrast, PPARα activation increased hepatic Acox, Fads1, Fads2 and Elovl5 mRNA levels, but reduced the DHA concentrations in the liver, retina, and phospholipid of brain cortex, and decreased mRNA and protein levels of the brain-derived neurotrophic factor in brain cortex. CONCLUSIONS: LCPUFA enzyme expression was altered by PPARα. Either PPARα deficiency or activation-decreased tissue DHA concentration is a stimulus for further studies to determine the functional significance.
Animals ; Brain ; Brain-Derived Neurotrophic Factor ; Clofibrate ; Docosahexaenoic Acids ; Fatty Acid Desaturases ; Liver ; Mice ; Peroxisomes ; PPAR alpha ; Retina ; RNA, Messenger

The aim of this paper was to screen the active targets of Schizonepetae Herba and Saposhnikoviae Radix in the treatment of ulcerative colitis by means of network pharmacology,and to investigate their mechanism of action. The effective components of Schizonepetae Herba and Saposhnikoviae Radix were screened out by traditional Chinese medicine systematic pharmacological( TCMSP)database,with oral bioavilability( OB) ≥30% and drug-like( DL) ≥18% selected as the thresholds. Target PPI network was built between the main components and their corresponding targets. One hundred and eighty-two human genes corresponding to the medicine target sites were obtained from Uniprot database; 3 874 genes corresponding to ulcerative colitis were obtained from Genecard database.A total of 115 intersection genes were screened from disease genes and medicine genes,and the PPI interaction analysis was conducted by using String tool. Disease-target PPI network was drawn by using Cytoscape software,and component-target-disease network was constructed. One hundred and eight nodes and 1 882 connections were found,and then Cytoscape software was used to merge the networks and filter the core network for gene GO function analysis and KEGG pathway enrichment analysis. The mechanism of Schizonepetae Herba and Saposhnikoviae Radix was then verified by animal experiment. Gene GO functional analysis suggested that biological process,molecular functions and cell components were involved,and it was found that ulcerative colitis might be related to transcription factor activity,and cytokine receptor binding,etc. Gene KEGG pathway enrichment analysis showed that the mechanism of ulcerative colitis might be associated with TNF and Toll-like receptors( TLRs) signaling pathway-mediated cytoinflammatory factors interleukin-1( IL-1) and interleukin-6( IL6). The possible mechanism of the effective components of Schizonepetae Herba and Saposhnikoviae Radix in treating ulcerative colitis might be related to intervening the cytokine receptor binding of TNF and TLRs signaling pathways,reducing the transcription of nuclear factor-kappaB( NF-κB),and inhibiting the secretion of intestinal inflammatory factors IL-1 and IL-6.
Animals ; Apiaceae/chemistry* ; Colitis, Ulcerative/drug therapy* ; Databases, Genetic ; Drugs, Chinese Herbal/therapeutic use* ; Humans ; Interleukins/metabolism* ; Lamiaceae/chemistry* ; Medicine, Chinese Traditional ; Phytotherapy ; Plant Roots/chemistry* ; Protein Interaction Mapping ; Signal Transduction ; Software ; Toll-Like Receptors/metabolism*

OBJECTIVETo investigate expression features and correlation of genes expression on MyD88-dependent signaling pathway in synovial membrane (SM) of progression of knee osteoarthritis (OA).

METHODSSixty Wistar rats were randomly divided into 6 groups, including blank group (N), false surgical group, model groups[2 weeks (2W), 4 weeks (4W), 8 weeks (8W) and 12 weeks (12W)], with 10 rats in each group. The models were established by using Hulth method. Control group was experienced no surgery, while false surgical group was only opened joint cavity and sutured. The SM samples was collected according to the time designed above. The relative expression quantity of MyD88, TLR4 and NF-κB was detected by Real-time PCR after the extraction of the total RNA and reverse transcription. The correlation analysis was obtained by SPSS.

RESULTSThere was no significant difference in each gene mRNA expression between false surgical and blank group(> 0.05), while enhanced expression was found in the model groups(<0.05). The correlation index among MyD88, TLR4 and NF-κB was 0.91 and 0.86 respectively, and had significant difference among them.

CONCLUSIONSPositively relative among MyD88, TLR4 and NF-κB played main role in TLR4/NF-κB signal passway, and could predicate the expression of other genes in the passway. It also could further provide the basis for clarify the pathologic mechanism of knee OA.

Objective:To investigate the HBV infection and analyze the related risk factors among Li minority in Baisha county,Hainan Province. Methods: A total of 1 595 individuals of Li minority in Baisha county,were enrolled by random sampling method from July 2014 to October 2015. Epidemiological data including baseline characteristics and risk factors were obtained. HBcAb was detected by chemiluminescence method. The difference in age between HBcAb positive and negative group was analyzed by t test. The effects of age,gender and related risk factors on HBcAb were analyzed by univariate chi-square test and multivariate Logistic regression. Results: The positive rate of HBcAb was 71. 8% (1 145/1 595) and no significant difference between male and female was observed(x2=0. 134,P=0. 715). The difference of HBV infection among age groups was statistically significant (F=540. 769,P＜0. 001). The HBV infection rate was 11. 9% in the 12-17 year group,which was significantly lower than the others. The rate in the 18-23 year group (28. 0% ) was significantly higher than that in the 12-17 year group,but significantly lower than the other groups (＞85% ). Multivariate Logistic regression analysis showed that alcohol consumption and tattoo were independent risk factors associated with HBV infection (x2=169. 833,P＜0. 001;x2=11. 354,P=0. 001). Conclusion: The rate of HBV infection of Li minority in Baisha county,Hainan Province is high. The age,alcohol consumption and tattoo are the independent risk factors for infection.

OBJECTIVE: To compare the correlation between musculoskeletal ultrasound features, dysfunction and X-ray findings in patients with knee osteoarthritis, and to analyze the pathological mechanism of soft tissue inflammation in knee osteoarthritis. METHODS: Cross-sectional method was performed in this research (Evidence level: III). The patients with knee osteoarthritis were collected according to the screening criteria from September 2016 to January 2017 in Orthopedic clinic in our hospital. Musculoskeletal ultrasound and X-ray images were obtained and measured, knee function was measured by Lysholm scale. Pearson coefficient, t test and Wilcoxon were applied to analyze the correlation between soft tissue inflammation, knee dysfunction and X-ray features. RESULTS: Total 123 patients with knee osteoarthritis were recruited in this research. Soft tissue inflammation around knee had a high incidence in patients with knee osteoarthritis (infrapatellar fat pad inflammation 81%), and the synovial membrane thickness, joint effusion depth and meniscus bulging were beyond the normal range. Correlation analysis showed that the about Lysholm score and joint effusion depth had negative correlations with "Squat" score(=-0.21, =0.02). and Medial meniscus bulging had negative correlations with "Sustain" score(=-0.26, <0.01) and Lysholm total score (=-0.19, =0.04). Lateral meniscus bulging had a negative correlation with "Unstable" score (=-0.22, =0.02). The X-ray features, and medial joint space narrow had negative correlations with joint effusion depth(=-0.27, <0.01) and synovial membrane thickness(=-0.17, =0.007), and had a positive correlation with medial meniscus bulging. Medial joint space narrow was significantly correlated with patellar ligament inflammation and fat pad inflammation(<0.05). Lateral joint space narrow was significantly correlated with patellar ligament inflammation(=0.02). CONCLUSIONS Soft tissue inflammation around the knee-a major pathological manifestation of knee osteoarthritis, has significant correlations with knee dysfunction and bony structure lesions, and affects the progression of knee osteoarthritis by damaging knee joint function and promoting the destruction of articular cartilage.
Cartilage, Articular ; Cross-Sectional Studies ; Humans ; Knee Joint ; Menisci, Tibial ; Osteoarthritis, Knee