ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage， as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides， iridoid glycosides， and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.

ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage， as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides， iridoid glycosides， and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.

Objective To investigate the clinical efficacy and safety of 1 565 nm non-ablative fractional laser combined with topical minoxidil and oral finasteride in the treatment of patients with androgenetic alopecia(AGA).Methods Seventy-five male AGA patients with Norwood-Hamilton classification grade Ⅱ-Ⅲ,were randomly assigned into three groups:the control group 1,the control group 2 and the experimental group,with 25 cases in each group.Patients in the control group 1 received topical 5%minoxidil(1 mL,twice daily).Patients in the control group 2 were treated with both topical 5%minoxidil and oral finasteride(1 mg,once daily).Patients in the experimental group received a combined therapy of 1 565 nm non-ablative fractional laser in addition to topical 5%minoxidil and oral finasteride.Hair overall efficacy was evaluated using a 7-point rating scale after 24 weeks of treatment.Hair diameter and density were measured using a dermoscope.Patient satisfaction was assessed post-treatment,and adverse reactions were recorded.Results The overall efficacy of hair in the experimental group was superior to the control group 1 and the control group 2.There were no significant differences in hair density and hair diameter before treatment between the three groups(P＞0.05).After treatment,hair diameter and density increased in all three groups compared to baseline values(P＜0.05),and the hair diameter and hair density of the experimental group were higher than those of the control group 1 and the control group 2(P＜0.05).Patient satisfaction in the experimental group was higher than that in the control group 1 and the control group 2(P＜0.05).Patients in the experimental group experienced tolerable pain and burning sensations during laser treatment,and the symptoms were self-alleviated within a few hours.There were no serious adverse reactions reported in any group.Conclusion The combination therapy of 1 565 nm non-ablative fractional laser,5%minoxidil,and finasteride demonstrates significantly better efficacy in the treatment of AGA than minoxidil and finasteride alone drug therapy.

PTH-related peptide (PTHrP) improves the bone marrow micro-environment to activate the bone-remodelling, but the coordinated regulation of PTHrP and transforming growth factor-β (TGFβ) signalling in TMJ-OA remains incompletely understood. We used disordered occlusion to establish model animals that recapitulate the ordinary clinical aetiology of TMJ-OA. Immunohistochemical and histological analyses revealed condylar fibrocartilage degeneration in model animals following disordered occlusion. TMJ-OA model animals administered intermittent PTHrP (iPTH) exhibited significantly decreased condylar cartilage degeneration. Micro-CT, histomorphometry, and Western Blot analyses disclosed that iPTH promoted subchondral bone formation in the TMJ-OA model animals. In addition, iPTH increased the number of osterix (OSX)-positive cells and osteocalcin (OCN)-positive cells in the subchondral bone marrow cavity. However, the number of osteoclasts was also increased by iPTH, indicating that subchondral bone volume increase was mainly due to the iPTH-mediated increase in the bone-formation ability of condylar subchondral bone. In vitro, PTHrP treatment increased condylar subchondral bone marrow-derived mesenchymal stem cell (SMSC) osteoblastic differentiation potential and upregulated the gene and protein expression of key regulators of osteogenesis. Furthermore, we found that PTHrP-PTH1R signalling inhibits TGFβ signalling during osteoblastic differentiation. Collectively, these data suggested that iPTH improves OA lesions by enhancing osteoblastic differentiation in subchondral bone and suppressing aberrant active TGFβ signalling. These findings indicated that PTHrP, which targets the TGFβ signalling pathway, may be an effective biological reagent to prevent and treat TMJ-OA in the clinic.
Animals ; Osteoclasts ; Osteogenesis ; Parathyroid Hormone-Related Protein/pharmacology* ; Temporomandibular Joint ; Transforming Growth Factor beta/pharmacology*

ObjectiveTo investigate the metabolites and gene expression characteristics in fibrous roots of Dioscorea zingiberensis in response to low phosphorus stress. MethodThe severe stress group， the moderate stress group， and the normal group were set up to stimulate the low phosphorus stress experiment. The fibrous roots of D. zingiberensis were collected during initial stress. The metabolites and transcriptomic characteristics were analyzed by gas chromatography-mass spectrometry （GC-MS） derivatization and RNA-seq techniques. Through multivariate statistical analysis of metabolites treated by different methods，functional analysis of differentially expressed genes， and data mining， the metabolism markers produced in fibrous roots of D. zingiberensis under low phosphorus stress were screened out， and the metabolic pathway characteristics of different genes were analyzed. ResultA total of 116 GC-MS metabolites were detected from the fibrous roots of D. zingiberensis. The metabolic characteristics of fibrous roots of D. zingiberensis under different low phosphorus treatments were obviously different. Orthogonal partial least squares discriminant analysis（OPLS-DA） model was used to screen six differential metabolites represented by sugars and alcohols from metabolites of fibrous roots treated with different methods，and these components were presumedly metabolism markers of fibrous roots of D. zingiberensis in response to low phosphorus stress. The differential genes screened out from the severe stress group and the normal group were mainly enriched in peroxidase pathway，phosphate and hypophosphate metabolism pathway，while the differential genes screened out from the severe stress group and the moderate stress group were mainly enriched in glutathione metabolism pathway and phosphopentose pathway. A total of 177 differential genes in response to low phosphorus stress were screened out from fibrous roots， involving many pathways such as terpenoid skeleton and inositol biosynthesis，which was consistent with the fact that the metabolic differential components in fibrous roots in response to low phosphorus stress were mainly saccharides and inositol. ConclusionThe metabolites and gene expression in fibrous roots of D. zingiberensis responded to low phosphorus stress，and the differential metabolites were closely related to differentially expressed genes. This study is expected to provide a theoretical basis for the research on the molecular mechanism of D. zingiberensis in response to low phosphorus stress.

ObjectiveTo analyze the effect of different fixation methods on the chemical quality characteristics and antioxidant activity of Rehmannia glutinosa leaves， so as to lay a foundation for the selection of processing technology and quality evaluation of this medicinal materials. MethodR. glutinosa leaves was dried at 55 ℃ after treating by three fixation methods （55 ℃， boiling water， 105 ℃）， and then the fingerprints of R. glutinosa leaves were collected by high performance liquid chromatography （HPLC） and near infrared spectroscopy （NIRS）， and their antioxidant activities were analyzed by the 2，2-biphenyl-1-picrylhydrazyl （DPPH） method. Finally， similarity analysis， principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA） and Pearson correlation analysis were used to comprehensively evaluate the quality of R. glutinosa leaves with different fixation methods. ResultThe results of HPLC and NIRS fingerprint analysis indicated that there were differences in the quality characteristics of R. glutinosa leaves by different fixation treatments. The comprehensive score of chemical quality of R. glutinosa leaves by fixation at 55 ℃ was the highest， its average comprehensive score was 2.096， followed by fixation at 105 ℃， and the lowest was fixation with boiling water. The antioxidant activity of sample with fixation at 55 ℃ was the highest， followed by fixation with boiling water. The results of OPLS-DA showed that verbascoside， isoacteoside and catalpol were the main components causing the difference in chemical quality of the leaves from the three treatments， and the three components were positively correlated with the antioxidant activity of R. glutinosa leaves. Among them， the correlation between verbascoside and antioxidant activity was extremely significant， and the isoacteoside was significant. ConclusionThe chemical quality and antioxidant activity of R. glutinosa leaves are affected by the method of fixation， and the fixation at low temperature is the best primary processing method of R. glutinosa leaves.

【Objective】 To investigate the unqualified rate of anti-HIV detection of blood screening laboratories in Beijing-Tianjin-Hebei region, and explore the differences in anti-HIV detection ability and influencing factors in each laboratory. 【Methods】 Through filling questionnaires via e-mail, the anti-HIV ELISA unqualified rate and confirmed (WB) positive results (data) from January to December 2018 from 15 blood screening laboratories in Beijing-Tianjin-Hebei region were collected. Our laboratory was responsible for data collection and confirmation, and statistics software SPSS22.0 was used for analysis. 【Results】 1) There was a statistically significant difference among the unqualified rate of anti-HIV ELISA(6.77‱~35.71‱) and confirmed positive rate(0.60‱~3.56‱) in 15 blood screening laboratories in Beijing-Tianjin-Hebei region (P<0.05); 2) There were significant differencse among the ELISA unqualified rate and the confirmed positive rate of 8 reagents for anti-HIV detection(P<0.01), and the sensitivity of the 4th generation detection reagent and the imported reagent was higher than that of the 3rd generation reagent and the domestic reagent. The anti-HIV ELISA unqualified rate of R5 was the highest (19.08‱). 3)There were significant differences in the anti-HIV ELISA unqualified rate of R1, R2, R3, R5 and R7 reagents among different blood station laboratories(P<0.05), and there were no significant differences in the anti-HIV ELISA unqualified rate of R4, R6 and R8 reagents among different blood station laboratories(P>0.05). 4)The unqualified rate of anti-HIV ELISA of laboratories using different regents showed significant differences(P<0.05), except H, J, M. The unqualified rate of imported reagent was significantly higher than that of domestic reagents of laboratories using imported and domestic reagents combinations(P<0.05), except O. 62.5% (5/8) laboratories using domestic 3^rd and 4^th generation reagent combination showed significant differences in the unqualified rates among different reagents(P<0.05); 5) The positive rate of single-reagent(62.02%~95.45%)in 15 blood screening laboratories showed significant difference(P<0.001), and A was the lowest (62.02%). 【Conclusion】 The anti-HIV detection ability among 15 blood screening laboratories in Beijing-Tianjin-Hebei region is quite different. The application of different reagents is the main factor for the difference, and other factors such as personnel, instruments and test strategies also has a great impact on the detection of anti-HIV. It is still necessary to promote the process of homogenization of blood testing quality among blood screening laboratories in Beijing-Tianjin-Hebei region.

Objective:To identify the expression level of known microRNA (miRNA) by high-throughput sequencing and analysis of the miRNA of Schistosoma japonicum schistosomula, and predict the miRNA target genes and their biological functions. Methods:Schistosoma japonicum schistosomula were prepared in vitro, and total RNA of schistosomula were extracted and analyzed to construct a library for performing high-throughput sequencing. The difference of miRNA expression was analyzed by using DEGseq R language package and perl script. Then the target genes and their biological functions of differential miRNA were predicted by miRanda software, Blast software, and KEGG database respectively. Results:There were 38 483 matching sequences in the miRNA expressed of Schistosoma japonicum schistomula in the constructed library compared with the latest miRBase database, and 60 miRNA were identified, sja-miR-125b was the miRNA with the highest expression, followed by sja-miR-61, sja-miR-71a, sja-miR-36-3p and sja-miR-10-5p, which accounted for 91% (3 263/3 585) of the total miRNA expression. MiRanda software predicted a total of 7 176 target genes, gene functions concentrated on nucleotide transferase activity, cellular nitrogen complex metabolism, molecular function, biological processes, biosynthesis, plasma membrane and protein maturation. The functional enrichment analysis showed that the highly expressed miRNA were mainly involved in pathogenic process, biological progress and multiple metabolic regulation pathways. Conclusions:The miRNA expressed significantly of Schistosoma japonicum schistomula mainly involved in the regulation of metabolic pathways during the differentiation, growth and pathogenesis of Schistosoma japonicum. To lay a foundation for the study of the regulatory mechanism of Schistosoma japonicum development and the development of new drugs.

Objective: To compare the carcinogenic abilities of CD133⁺CD44⁺ laryngeal cancer stem cells and general laryngeal cancer stem cells and to identify the mechanism underlying the action of miRNAs. Methods: Solid tumor-derived laryngeal carcinoma stem cells and Hep-2-derived laryngeal carcinoma stem cells were cultured, and CD133⁺CD44⁺ laryngeal cancer stem cells were sorted by flow cytometry. Boden chamber invasion assay, cell migration assay and tumor formation assay were then performed to compare the invasion, migration and tumorigenic abilities of CD133⁺CD44⁺ laryngeal cancer stem cells and general laryngeal cancer stem cells. And then, miRNAs isolated from two laryngeal cancer stem cells were detected and analysed with miRNA chip. Results: (1)In Boyden chamber invasion assay, the cell invasion rate of CD133⁺CD44⁺ laryngeal cancer stem cells was obviously higher (80.2%±2.3% vs. 63.9%±3.2%, t=5.011, P=0.027); (2)CD133⁺CD44⁺ laryngeal cancer stem cells also had higher mobility in cell migration assay (82.9%±1.1% vs. 70.9%±0.6%, t=4.514, P=0.031); (3)In tumor formation assay, the tumor formation rate of CD133⁺CD44⁺ laryngeal cancer stem cells was also higher (80% vs. 50%). What′s more, we identified 15 miRNAs that were significantly upregulated in CD133⁺CD44⁺ laryngeal cancer stem cells and 3 miRNAs that were significantly downregulated in CD133⁺CD44⁺ laryngeal cancer stem cells, compared with normal laryngeal cancer stem cells. Conclusions CD133⁺CD44⁺ laryngeal cancer stem cells have stronger invasion, migration and tumorigenic abilities compared with normal laryngeal cancer stem cells, and the difference of miRNAs′ expression is one of the possible causes.

Objective To evaluate the effect of different doses of Astragalus membranaceus on the levels of vascular endothelial growth factor ( VEGF) and basic fibroblast growth factor ( bFGF) in serum and lung tissues of rats with pulmonary embolism. Methods Seventy-six clean-grade healthy male Sprague-Dawley rats, aged 8-9 weeks, weighing 140-170 g, were assigned to control group ( group C, n=11) and experimental group ( group E, n=65) by a random number table method. The rats with pulmonary em-bolism in group E were further divided into 4 subgroups using a random number table method: pulmonary embolism group (group P), low-dose Astragalus membranaceus group (group H1), median-dose Astraga-lus membranaceus group ( group H2 ) and high-dose Astragalus membranaceus group ( group H3 ) . The model of pulmonary embolism was established by injecting autologous blood clots into the right jugular vein. At 1 h and 1, 2, 3, 4, 5 and 6 days after successful establishment of the model, Astragalus membrana-ceus 20, 40 and 60 g∕kg were injected intraperitoneally in H1-3 groups, respectively, while the equal vol-ume of normal saline was given instead in group P. The chest was opened after anesthesia on day 7, and blood samples were collected from cardiac chambers for determination of concentrations of serum VEGF and bFGF by enzyme-linked immunosorbent assay. The pulmonary specimens were obtained from the upper lobe of right lungs for determination of the expression of VEGF and bFGF mRNA ( using real-time polymerase chain reaction) . Results Compared with group C, the concentrations of serum VEGF and bFGF were sig-nificantly increased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in the other four groups (P<0. 05). Compared with group P, the serum bFGF concentration was significantly in-creased, and the expression of VEGF and bFGF mRNA in lung tissues was up-regulated in H1-3 groups ( P<0. 05) . Compared with group H1, the serum bFGF concentration was significantly increased, the ex-pression of VEGF mRNA and bFGF mRNA in lung tissues was up-regulated in H2 and H3 groups ( P<0. 05) . Compared with group H2, the expression of VEGF and bFGF mRNA in lung tissues was significant-ly up-regulated in group H3 ( P<0. 05 ) . Conclusion Astragalus membranaceus can up-regulate the ex-pression of VEGF and bFGF in lung tissues in a dose-dependent manner, thus improving pulmonary embol-ism in rats.