BACKGROUND: Pemphigus is an autoimmune bullous disease with circulating desmosomal autoantibodies of IgG. In direct IF studies with perilesional tissue, IgA or IgM antibodies can be seen in addition to IgG. OBJECTIVE: We examined sera of patients with pemphigus for the presence/frequency of IgA autoantibodies as well as IgG by indirect IF and immunoblot assay. Patients: Twenty patients of pemphigus (PV 10, PF 10) who showed positive findings in indirect IF examinations. METHODS: Indirect IF study with normal human skin substrates and immunoblot analysis using A431 cell extracts (with multi-step immunostaining) were performed with patients sera. RESULTS: In indirect IF, IgA autoantibodies that bind to the epidermal keratinocyte antigens were detected in 4 cases among the 20 patients (PV 2 and PF 2). In immunoblot analysis IgA bands reacting to PV/PF antigens were observed in 7 cases from the 20 patients with pemphigus (PV 3, PF 4). The serum titers of IgA autoantibodies were lower than those of IgG in every single case. CONCLUSION: In patients with pemphigus (PV/PF), 35% of cases have serum IgA autoantibodies as well as IgG autoantibodies specific to the pemphigus antigens (Dsg 1/Dsg 3). However, pathogenic roles of the associated IgA autoantibody are not clear.
Antibodies ; Autoantibodies ; Cell Extracts ; Humans ; Immunoglobulin A* ; Immunoglobulin G ; Immunoglobulin M ; Keratinocytes ; Pemphigus* ; Skin

OBJECTIVETo identify the active material of anti-hepatic fibrosis from Amydae Carapax.

METHODSMembrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer.

RESULTSProteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components.

CONCLUSIONThree active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.

Animals ; Cell Line ; Humans ; Liver Cirrhosis ; Medicine, Chinese Traditional ; Tissue Extracts ; isolation & purification ; pharmacology

This research is to investigate study the flavonoids from stems of Nelumbo nucifera and the cytotoxic activities of iso- lated compounds. The constituents were separated by column chromatography,and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for cytoxic activities by MTT method. Twelve compounds were isolated and identified as rhamnazin-3-O-beta-D-glucopyranoside (1), luteolin-3', 4'-dimethylether-7-O-beta-D-glucoside (2), kaempferol-3-O-beta-D-xylopyranosyl-(1-->2)-O-beta-D-glucopyranoside (3), quercetin-3,3'-di-O-beta-D-glucopyranoside (4), 1, 8-dihydroxy-3,7-dimethoxyxanthone (5), isorhamnetin-3-O-beta-D-glucopyranoside(6) , kaempferol(7), isorhamnetin (8), quercetin(9), astragalin(10), hyperoside (11) and 1-hy- droxy-3,7,8-trimethoxyxanthone(12). All compounds were isolated from stems of this plant for the first time, and compounds 1-5 were firstly isolated from the family nelumbonaceae. Compounds 24 and 6 showed significant cytotoxic activities against BEL-7402 carcinoma cell lines at a concentration of 1 x 10(-5) mol x L(-1) with the inhibitory rate of 67.36%, 53.25%, 57.78%, 60.13% and 52.11%, respectively.
Cell Line, Tumor ; Flavonoids ; chemistry ; pharmacology ; Humans ; Nelumbo ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Plant Stems ; chemistry

OBJECTIVETo investigate the chemical constituents of Excoecaria acerifclia and their antitumor activities.

METHODThe constituents were isolated and purified by column chromatography. Their structures were identified by their physicochemical properties and spectral features. Cytotoxicities of the purified compounds were evaluated by MTI method against human cancer cell lines HepG2.

RESULTEight compounds were isolated and identified as: 7-hydroxy-6-methoxy-coumarin (1), 8-hydroxy-6,7-methoxy-coumarin (2), kaempferol (3), kaempferol-3-O-beta-D-glucoside (4), quercetin-3-O-beta-D galactoside (5), kaempferol-3-O-beta-D-glucoside-2"-gallate (6), beta-sitosterol (7), daucosterol (8).

CONCLUSIONAll compounds were isolated from this plant for the first time. Compounds 5 showed inhibitory activity towards HepG2 with IC50 values of 7.13 mol x L(-1).

Cell Proliferation ; drug effects ; Euphorbiaceae ; chemistry ; Hep G2 Cells ; Humans ; Plant Extracts ; analysis ; isolation & purification ; pharmacology

Thirty-three compounds were isolated from the root decoction of Isatis indigotica by using a combination of various chromatographic techniques including silica gel, macroporous adsorbent resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by spectroscopic data as (+)-dehydrovomifoliol (1), (S)-(+)-abscisic acid (2), vomifoliol (3), cyclo (L-Phe-L-Leu) (4), cyclo(L-Phe-L-Tyr) (5), cyclo(L-Tyr-L-Leu) (6), cyclo(L-Pro-L-Tyr) (7), evofolin B (8), (+)-syringaresinol (9), (-)-(7R,7'R,8S,8'S)-4,4'-dihydroxy-3-methoxy-7,9';7',9-diepoxy-lignan (10), (-)-medioresinol (11), (+) -(7R,7'R,8S,8'S) -neo-olivil (12), (-) -5-methoxyisolariciresinol (13), 1,3-dihydro-2H-indol-2-one (14), isalexin (15), dihydroneoascorbigen (16), indican (17), (-) -(S) -cyanomethyl-3-hydroxyoxindole (18), isoformononetein (19), calycosin (20), stigamast-5-ene-3beta-ol-7-one (21), acetovanillone (22), 3, 5-dimethoxy-4-hydroxyacetophenone (23), dihydroconiferyl alcohol (24), dihyroferulic acid (25), 3-hydroxy-1-(4-hydroxyphenyl) propan-1-one (26), beta-hydroxypropiovanillone (27), 4-aminobenzoic acid (28), 3-(4-hydroxyphenyl) propan-1-ol (29), 4-(2-hydroxyethyl) phenol (30), 2-methoxy-4-vinylphenol (31), pyrocatechol (32), and 4-pentenamide (33). These compounds were isolated from the root of I. indigotica for the first time. In preliminary in vitro assays, compound 19 showed activity against the influenza virus A/Hanfang/359/95 (H3N2), the herpes simplex virus 1 (HSV-1), and Coxsackie virus B3 (Cox-B3), with IC50 values of 2.06, 6.84, and 8.70 micromol x L(-1), respectively, but other compounds were in-active at a concentration of 1.0 x 10 x (-5) mol x L(-1).
Animals ; Cell Line ; Humans ; Isatis ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; toxicity ; Plant Roots ; chemistry

Nineteen compounds, including kihadanin D (1), obacunone (2), kihadanin A (3), kihadanin B (4), kihadanin C (5), limonin (6), evodol (7), fraxinellone (8), furo[2,3-b]quinolin-4-ol (9), preskimmianine (10), ifflaiamine (11), dictamnol (12), naringenin (13), diosmetin (14), wogonin (15), scopoletin (16), cleomiscosin A (17), apocynin (18), and methyl pyroglutamate (19), were isolated from the methanol extract of the root barks of Dictamnus dasycarpus by using various column chromatographies. Their chemical structures were extensively determined on basis of UV, IR, NMR, MS, and CD spectroscopic data analyses. Among them, 1 is a new limonoid, 9 was isolated from plant kingdom for the first time, 11, 13-14 and 17-19 were obtained from the genus Dictamnnus for the first time. Cytotoxicities of compounds 1-18 were tested, and the results indicated that 1 exhibited cytotoxicities against three human cancer cell lines MDA-MB-231, A549 and HT29 with IC₅₈ values of 16.22, 21.72 and 31.06 μmol·L⁻¹, respectively.
Cell Line, Tumor ; Dictamnus ; Humans ; Molecular Structure ; Plant Bark ; Plant Extracts ; Plant Roots

Ischemic cerebrovascular disease and cerebral ischemia/reperfusion injury threaten the health of human being. We studied the protective effect of Ginkgo biloba extract 50 (EGb50) on the mitochondrial function in SH-SY5Y cells after hypoxia/reoxygenation (H/R) injury and explored its mechanisms, so as to provide new ideas for studies on the treatment for ischemic cerebrovascular disease. We established the H/R injury model in SH-SY5Y cells after administrating EGb50. Subsequently, the mitochondrial membrane potential and the concentration of intracellular Ca²⁺ were measured by flow cytometer. The levels of optic atrophy1 (Opa1) and dynamin-like protein 1 (Drp1) were evaluated by immunofluorescence and western blot. The results showed that the mitochondrial membrane potential was decreased and the level of intracellular Ca²⁺ was increased after H/R injury. Moreover, the expression of mitochondrial fusion protein Opa1 was decreased, while the expression of mitochondrial fission protein Drp1 was increased. However, EGb50 significantly increased the mitochondrial membrane potential and suppressed the level of intracellular Ca²⁺. In addition, EGb50 increased the expression of Opa1 and decreased the expression of Drp1. The results demonstrated that EGb50 has a neuroprotective effect on SH-SY5Y cells after H/R injury, and could improve the energy metabolism and mitochondrial function. The underlying mechanisms may be associated with the regulation of mitochondrial fusion and fission, which provided data support for the treatment of ischemic cerebrovascular disease with EGb50.
Cell Hypoxia ; Humans ; Membrane Potential, Mitochondrial ; Mitochondria ; Plant Extracts ; Reperfusion Injury

BACKGROUND AND OBJECTIVES: We considered two possible mechanisms that might be responsible for the increased accumulation of lysozyme in retinoic acid (RA)-deficient cultures, either increased lysozyme synthesis or decreased lysozyme degradation based on our previous data. This study was to determine whether the synthesis and decay rate of intracellular lysozyme in RA-sufficient cultures are different from those in RA-deficient cultures. MATERIALS AND METHOD: Passage-2 normal human airway epithelial cells were used. For synthesis rate of lysozyme, day 10 RA-deficient and RA-sufficient cultures, incubated over 6 hour period with 35S-methionine-cysteine and cell lysates, were collected. For decay rate, day 10 cultures grown in the presence or absence of RA were labeled with 35S-methionine-cysteine for 4 hours and the labeling media were then removed. Cell extracts were collected over 8 hours. Newly synthesized or labeled lysozyme was immunoprecipitated with anti-lysozyme antibody and separated by SDS-PAGE. RESULTS: Lysozyme synthesis rate in RA-sufficient cultures was higher than in RA-deficient cultures. In the RA-deficient cultures, the levels of newly synthesized lysozyme barely changed over the 8 hour post-labeling period. In contrast, in the RA-sufficient cultures, radiolabeled lysozyme levels decreased rapidly during the 8 hour post-labeling period, with a half-life of approximately 6 hours. CONCLUSION: Discrepancy in mRNA and protein of lysozyme in RA-deficient cultures is due to the increased stability of lysozyme protein in RA-deficient cultures.
Cell Extracts ; Electrophoresis, Polyacrylamide Gel ; Epithelial Cells* ; Half-Life ; Humans* ; Muramidase* ; RNA, Messenger ; Tretinoin*

Andrographis paniculata (Burm. f.) Nees (AP) is commonly used for the treatment of many infectious diseases and has been cultivated widely in Asian countries, and has been included in United States Pharmacopoeia as a dietary supplement, but the cultivars of A. paniculata are not abundant due to its self-pollinated. With the aims to enrich AP resources and provide materials for after breeding we explored the polyploidy induction. Different explants, colchicine concentration, and treatment time were tested. After identification by flow cytometry, eleven polyploid plants with different morphologic traits were obtained. The agronomic traits and andrographolide concentration of the polyploids were improved greatly. One of the polyploids (serial 3-7) was chosen for further study. The traits of the second and third generation polyploids (serial 3-7) were stable. Compared with the normal plants, the seeds (2nd generation) weight increased by 31%, and the andrographolide concentration of the leaves increased by 14% (2nd) and 28% (3rd). In conclusion, AP autopolyploids with different morphologic traits were established successfully for the first time, and the polyploids induction might be effective for crop improvement of AP.
Andrographis ; chemistry ; genetics ; growth & development ; Breeding ; Cell Culture Techniques ; Plant Extracts ; chemistry ; Polyploidy

OBJECTIVE: To explore the antitumor effects of ethanol extract from Ventilago leiocarpa Benth (EEVLB) on sarcoma 180 (S180) tumor-bearing mice and the potential mechanism. METHODS: Sixty mice were randomly assigned to 6 groups according to a random number table: normal group, model group, 5-fluorouracil (5-FU) group (0.02 g·kg RESULTS: EEVLB with different concentrations achieved inhibition of tumor growth in vivo, wherein the high-dose group showed the most significant reduction in tumor weight and increased apoptosis of tumor cells (P<0.05). In addition, both net weight gain and spleen index of mice showed uptrend in EEVLB treatment groups (P<0.05). Besides, serum levels of IL-2 and IL-6, percentages of CD3 CONCLUSIONS EEVLB exhibits promising antitumor activity in vivo. This effect might be due to activation of apoptotic signaling pathway, increase of cytokine levels and enhancement of immune function in tumor-bearing mice.
Animals ; Cell Line, Tumor ; Ethanol ; Mice ; Plant Extracts/therapeutic use* ; Rhamnaceae ; Sarcoma 180/drug therapy*