1.Alkaloids isolated from Buyang Huanwu Decoction and their structure identification
Jun WU ; Pengfei TU ; Yuying ZHAO
Chinese Traditional and Herbal Drugs 1994;0(07):-
Objective To study the antithrombotic compounds and bioactive fraction from Buyang(Huanwu) Decoction(BHD).Methods Four alkaloids have been isolated from BHD by repeated chromatographies.Their structures were elucidated by means of spectral and single-crystal Xray diffraction analysis.Results They are N-(3′-maleimidyl)-5-hydroxymethyl-2-pyrrole formaldehyde(Ⅰ),4-carbamoyl-2-pyrrole carboxylic acid(Ⅱ),N-(1′-D-deoxyxylitolyl)-6,7-dimethyl-1,4-dihydro-2,3-quinoxalinedione(Ⅲ),and 2,3,4,9-tetrahydro-1H-pyrido indole-3-carboxylic acid(Ⅳ).Conclusion All these compounds are isolated from BHD for the first time and compounds Ⅰ-Ⅲ are new alkaloids.
2.Isolation and identification of four isoflavones and one pterocarpan from BUYANGHUANWUTANG
Jun WU ; Pengfei TU ; Yuying ZHAO
Chinese Traditional and Herbal Drugs 1994;0(07):-
Object To study the antithombotic portion and bioactive fraction of BUYANGHUANWUTANG (BHT), a decoction of astragalus in combination with peony and several other ingredients. Methods The EtOAc portion of BHT extract was repeatedly separated by chromatography and identified by means of spectral analysis. Results One pterocarpan, (6aR, 11aR) 9, 10-dimethoxypterocarpan-3-O-?-D-glucopyanoside (Ⅴ) and four isoflavones, formononetin (Ⅰ), calycosin (Ⅱ), formononetin-7-O-?-D-glucopyanoside (Ⅲ), and calycosin-7-O-?-D-glucopyranoside (Ⅳ) were identified. Conclusion All five compounds were isolated for the first time from BHT.
3.Isomers of amygdalin in BUYANG HUANWU TANG and its production
Jun WU ; Pengfei TU ; Yuying ZHAO ;
Chinese Traditional and Herbal Drugs 1994;0(02):-
Object To study the isomers of amygdalin in BUYANG HUANWU TANG and its production Methods Amygdalin was isolated from both peach seeds and BUYANG HUANWU TANG by using various column chromatography Their structures were identified by the various spectral data Results Amygdalin had been isolated from n BuOH fraction of aqueous extract of BUYANG HUANWU TANG and found to be a pair of D , L epimers and their ration was 1∶1 It was also found that the structures and the ration of D , L epimers of amygdalin in decoction of single peach seed were similar to that in BUYANG HUANWU TANG. The peach seed only gave D amygdalin when it was extracted in 95% EtOH at reflux temperature, and D amygdalin cannot be isomerized when it was treated in water at 100 ℃ Conclusion Isomerization of D amygdalin results from interaction between it and other compounds of peach seed in water at high temperature, and has no evident relation to other constituents in BUYANG HUANWU TANG L amygdalin is a new compound generated due to decoction of peach seed
4.Clinical analysis of 34 cases of bullous pemphigoid with non-bullous lesions as initial manifestation
Caihong SUN ; Hongqin TU ; Yuying KANG ; Mei JU ; Baozhu CHANG ; Heng GU
Chinese Journal of Dermatology 2008;41(10):647-649
Objective To analyse the clinical and therapeutic features as well as laboratory findings in bullous pemphigoid with non-bullous lesions as initial manifestation. Methods Clinical data on 34 cases of bullous pemphigoid with non-bullous lesions as initial manifestation were retrospectively analyzed. Results The male to female ratio was 1.83 :1, with a mean age of onset at 59.79±15.63 years. Before typical bullae appeared, patients presented with erythema, papules, papulovesicles' plaques" wheals, nodules,or erythema muitiforme-like lesions, with the most common lesions being erythematous papules and plaques (occumng in 35.29% of these patients). Conclusions Among these patients, nearly 1/3 displayed various skin lesions at the onset; simultaneous erythematous papules and plaques are the most common initial manifestation.
5.Cloning and expression of Chlamydia trachomatis heat shock protein 60 gene
Juanhua LIU ; Mukai CHEN ; Qiman LIAO ; Haipian LI ; Yuying TU ; Jiande HAN
Chinese Journal of Dermatology 2009;42(5):318-320
Objective To clone and express Chlamydia trachomatis (Ct) heat shock protein 60 (hsp60) gene. Methods The hsp60 gene fragment was amplified from Ct chromosomal DNA by PCR. After purification and digestion with enzymes Sal I and Not I , the hsp60 gene fragment was inserted into the compatible site of prokaryotic expression vector pET-28a. The constructed recombinant plasmid was identified by PCR, restriction enzyme cleavage and sequencing, then, it was transfected into an expression strain Escherichia coli BL21 (DE3). The expression of fusion protein was induced by isopropy-β-D- thiogalactoside (IPTG) in the host bacteria, and the expressed product was identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blot. Results PCR and restriction enzymes cleavage analysis confirmed that the hsp60 gene was successfully cloned into the recombinant plasmid. DNA sequencing showed that the sequence of cloned gene was fully consistent with the published sequence in Genebank. As revealed by SDS-PAGE, the size of expressed fusion protein approximated 60 kilodaltons, and Western-blot confirmed the expressed product to be the expected protein. The final concentration of fusion protein was 17.85 mg/L with a purity of more than 90%. Conclusions A recombinant expression plasmid pET-28a-hsp60 is successfully constructed in this study, and soluble hsp60 protein is expressed by the recombinant plasmid-transfected E. coli.
6.Effects of VX765 on osteoarthritis and chondrocyte inflammation in rats.
Wanran HUANG ; Junxue TU ; Aiqing QIAO ; Chujun HE
Chinese Journal of Reparative and Reconstructive Surgery 2024;38(1):74-81
OBJECTIVE:
To investigate the effects and underlying mechanisms of VX765 on osteoarthritis (OA) and chondrocytes inflammation in rats.
METHODS:
Chondrocytes were isolated from the knee joints of 4-week-old Sprague Dawley (SD) rats. The third-generation cells were subjected to cell counting kit 8 (CCK-8) analysis to assess the impact of various concentrations (0, 1, 5, 10, 20, 50, 100 μmol/L) of VX765 on rat chondrocyte activity. An in vitro lipopolysaccharide (LPS) induced cell inflammation model was employed, dividing cells into control group, LPS group, VX765 concentration 1 group and VX765 concentration 2 group without obvious cytotoxicity. Western blot, real-time fluorescence quantitative PCR, and ELISA were conducted to measure the expression levels of inflammatory factors-transforming growth factor β 1 (TGF-β 1), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α). Additionally, Western blot and immunofluorescence staining were employed to assess the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1). Thirty-two SD rats were randomly assigned to sham surgery group (group A), OA group (group B), OA+VX765 (50 mg/kg) group (group C), and OA+VX765 (100 mg/kg) group (group D), with 8 rats in each group. Group A underwent a sham operation with a medial incision, while groups B to D underwent additional transverse incisions to the medial collateral ligament and anterior cruciate ligament, with removal of the medial meniscus. One week post-surgery, groups C and D were orally administered 50 mg/kg and 100 mg/kg VX765, respectively, while groups A and B received an equivalent volume of saline. Histopathological examination using HE and safranin-fast green staining was performed, and Mankin scoring was utilized for evaluation. Immunohistochemical staining technique was employed to analyze the expressions of matrix metalloproteinase 13 (MMP-13) and collagen type Ⅱ.
RESULTS:
The CCK-8 assay indicated a significant decrease in cell viability at VX765 concentrations exceeding 10 μmol/L ( P<0.05), so 4 μmol/L and 8 μmol/L VX765 without obvious cytotoxicity were selected for subsequent experiments. Following LPS induction, the expressions of TGF-β 1, IL-6, and TNF-α in cells significantly increased when compared with the control group ( P<0.05). However, intervention with 4 μmol/L and 8 μmol/L VX765 led to a significant decrease in expression compared to the LPS group ( P<0.05). Western blot and immunofluorescence staining demonstrated a significant upregulation of Nrf2 pathway-related molecules Nrf2 and HO-1 protein expressions by VX765 ( P<0.05), indicating Nrf2 pathway activation. Histopathological examination of rat knee joint tissues and immunohistochemical staining revealed that, compared to group B, treatment with VX765 in groups C and D improved joint structural damage in rat OA, alleviated inflammatory reactions, downregulated MMP-13 expression, and increased collagen type Ⅱ expression.
CONCLUSION
VX765 can improve rat OA and reduce chondrocyte inflammation, possibly through the activation of the Nrf2 pathway.
Rats
;
Animals
;
Chondrocytes/metabolism*
;
Matrix Metalloproteinase 13/metabolism*
;
Rats, Sprague-Dawley
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Tumor Necrosis Factor-alpha/metabolism*
;
Collagen Type II/metabolism*
;
Interleukin-6
;
Lipopolysaccharides/pharmacology*
;
NF-E2-Related Factor 2/pharmacology*
;
Inflammation/drug therapy*
;
Osteoarthritis/metabolism*
;
Transforming Growth Factor beta1/metabolism*
;
Dipeptides
;
para-Aminobenzoates
7.Construction and validation of a visual prediction model for the risk of urinary tract infection after PKRP surgery
Fan YE ; Yuying WAN ; Ping TU ; Chuntao XU
China Modern Doctor 2024;62(12):19-23
Objective To analyze the influencing factors of postoperative urinary tract infection in patients undergoing transurethral resection of the prostate with plasmakinetic energy(PKRP)and establish a risk prediction nomogram model.Methods The data of PKRP patients in Department of Urology,the Second Affiliated Hospital of Nanchang University from December 2020 to September 2021 were selected as the modeling set,and the high-risk factors were screened by univariate analysis and Logistic regression analysis.The risk prediction nomogram model was constructed and verified internally and externally.Results The incidence of urinary tract infection after PKRP surgery was 15.38%.Multivariate analysis showed that age,other location infection,diabetes,preoperative catheterization,urethral injury,indwelling catheter material,hair coloring catheter replacement times and number of indwelling catheterization were risk factors for urinary tract infection(P<0.05).Internal verification(area under the curve was 0.875)and external verification(area under the curve was 0.869)show that the risk prediction nomogram model has good discrimination and accuracy.Conclusion The influencing factors of urinary tract infection after PKRP are complex.The risk prediction nomogram model has good prediction performance,which can provide a basis for the prevention and treatment of urinary tract infection after PKRP.
8.Reactive oxygen species and nitric oxide scavenging nanoparticles alleviating rheumatoid arthritis through adjusting the seeds and growing soils.
Peng HUA ; Ruifeng LIANG ; Yanbei TU ; Yuying YIN ; Man-Kay LAW ; Meiwan CHEN
Acta Pharmaceutica Sinica B 2023;13(12):5016-5029
Normalizing inflamed soils including reactive oxygen species (ROS), nitric oxide (NO), cell-free DNA, and regulating inflammation-related seeds such as macrophages, neutrophils, fibroblasts, represent a promising strategy to maintain synovial tissue homeostasis for rheumatoid arthritis (RA) treatment. Herein, ROS scavenging amphiphilic block copolymer PEGylated bilirubin and NO-scavenging PEGylated o-phenylenediamine were fabricated to self-assemble into a dually responsive nanoparticle loaded with JAK inhibitor notopterol (Not@BR/oPDA-PEG, NBOP NPs). The simultaneous ROS and NO depletion combined with JAK-STAT pathway inhibition could not only promote M2 polarization to reduce further ROS and NO generation, but also decrease cytokines and chemokines to prevent immune cell recruitment. Specifically, NBOP NPs responded to high level ROS and NO, and disintegrated to release notopterol in inflamed joints as the hydrophobic heads BR and oPDA were transformed into hydrophilic ones. The released notopterol could inhibit the JAK-STAT pathway of inflammatory cells to reduce the secretion of pro-inflammatory cytokines and chemokines. This strategy represented an effective way to regulate RA soils and seeds through breaking the positive feedback loop of inflammation aggravation, achieving an excellent anti-RA efficacy in a collagen-induced arthritis rat model. Taken together, our work offered a reference to adjust RA soils and seeds for enhanced RA treatment.