Iridoid glycosides were the main active ingredient of Rehmannia glutinosa, of which catalpol has the highest content. This work will provide theoretical basis for metabolic study and cultivation of iridoids on the basis of the dynamic accumulation of catalpol and total iridoids in the growth of R. glutinosa. The samples of rehmannia 85-5 were gathered in the same filed from July to October. The contents of catalpol and total iridoid glycosides were measured by HPLC and specteophotometric, respectively. The results showed that youngest leaves had the higher content of catalpol and total iridoid glyosides than that of the other two leaf ages in the same growth stage from July to September, while their content of catalpol and total iridoid glycosides were all decreased as the growth of leaves of R. glutinosa. The content of catalpol didn't differ significantly from July to September, whereas it has significantly increased in October in the three leaf stage. In the same stage, the wider the root diameter is, the higher content of the effective components are. In August and September, the total iridoid glycosides have the fastest accumulation. The content of catalpol was increased as the accumulation of total iridoid glycosides.
Iridoid Glucosides ; metabolism ; Iridoids ; metabolism ; Plant Roots ; metabolism ; Rehmannia ; growth & development ; metabolism ; Seasons ; Water ; metabolism

The paper is aimed to study the enzymatic hydrolysis of the activatable cell-penetrating peptide (ACPP) that was designed and synthesized. The ACPP was composed of three parts, polyanionic sequence peptide, peptide sequence that specifically cleaved by matrix metalloproteinase (MMP) and cell penetrating peptide (CPP). The ACPP was hydrolyzed by type IV collagenase (MMP-2/9) under the condition of 37 degrees C and was monitored by reversed-phase high performance liquid chromatography (RP-HPLC). The efflux of peak was collected and detected by matrix assisted laser desorption ionization orthogonal time of flight mass spectrometry (MALDIO-TOF-MS) to speculate the sequences of the peptide fragments. The results indicated that the ACPP could be cleaved by type IV collagenase at target site as predicted, released CPP. The half life of the cleavage was about 4 h. Meanwhile, the peptide fragments may be cleaved again at other sites by type IV collagenase.
Amino Acid Sequence ; Cell-Penetrating Peptides ; chemical synthesis ; chemistry ; Chromatography, High Pressure Liquid ; Hydrolysis ; Matrix Metalloproteinase 2 ; chemistry ; Matrix Metalloproteinase 9 ; chemistry ; Peptide Fragments ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo confirm whether human MHC class I chain-related A (MICA) induces the amplification of V delta 1 gamma delta tumor-infiltrating lymphocytes (TILs) in vitro and to identify the cytotoxicity of MICA-reactive V delta 1 gamma delta TILs towards epithelial tumor cells.

METHODSMICA protein was prokaryoticly expressed and purified by molecular cloning technology. The purified recombined MICA (rMICA) was used to induce V delta 1 gamma delta T cells from tumor tissues in vitro and the cytotoxicity of these V delta 1 gamma delta TILs were tested by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT).

RESULTSThe rMICA was expressed in prokaryocyte with pET30 as a vector. The immobilized rMICA protein could markedly induce the amplification of V delta 1 gamma delta T cells from tumor tissue in vitro. These V delta 1 gamma delta T cells showed strong cytolytic activities towards tumor cell lines expressing MICA.

CONCLUSIONThe MICA-reactive V delta 1 gamma delta T cell may be a candidate for adoptive cellular therapy of tumors.

Adult ; Aged ; Female ; HeLa Cells ; pathology ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Humans ; Immunotherapy, Adoptive ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Middle Aged ; Ovarian Neoplasms ; immunology ; Receptors, Antigen, T-Cell, gamma-delta ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; T-Lymphocytes, Cytotoxic

AIMTo investigate the pharmacokinetics of doxorubicin alginate microspheres (DOX-AM) in vivo after hepatic arterial embolization.

METHODSChina miniature pigs were chosen as the experimental animals. Transcatheter hepatic arterial chemoembolization (TACE) with DOX-AM (experimental group), lipiodol and DOX (DOX-lipiodol, control group 1), and infusion with DOX (control group 2) were performed after angiography and superselection of an intrahepatic branch of hepatic artery. After chemoembolization or infusion, the blood was collected at different time intervals. Drug concentration in plasma was measured by HLPC and the parameters of pharmacokinetics were calculated.

RESULTSThe values of T1/2, AUC, Cmax, and MRT of the DOX-AM were significantly different from those of control group 1 and control group 2. After embolization, the DOX-AM embolized in the vessel and still retained there at 8 weeks. The digital subtraction arteriography (DSA) and computerized tomography (CT) showed the reliable embolization results. The histological examination indicated that the liver damnifications were changed transitorily in all groups (P < 0.05) and were recovered within two weeks. The liver damnifications increased in following order: DOX < DOX-AM < DOX-lipiodol.

CONCLUSIONDOX-AM showed definite property of delayed release of drug in liver, and increased the retention time and concentration of DOX after embolization in vivo.

Alginates ; chemistry ; Angiography, Digital Subtraction ; Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacokinetics ; Area Under Curve ; Chemoembolization, Therapeutic ; Doxorubicin ; administration & dosage ; blood ; pharmacokinetics ; Drug Carriers ; Female ; Hepatic Artery ; diagnostic imaging ; metabolism ; Iodized Oil ; chemistry ; Liver ; blood supply ; diagnostic imaging ; metabolism ; Male ; Microspheres ; Particle Size ; Swine ; Swine, Miniature ; Tomography, X-Ray Computed

OBJECTIVETo detect the expression of the glucocorticoid receptor-alpha/beta (GR-alpha/beta) mRNA in nasal polyp and normal nasal mucosa and investigate the significance of receptor-alpha/beta in GC-insensitive and GC-sensitive patients.

METHODSThe expression of GR-alpha/beta mRNA in 40 samples of nasal polyp and 30 normal nasal mucosa was examined by using Fluorescent quantitative PCR.

RESULTSForty cases were screened from the 80 cases after follow-up, including 26 cases of GC-sensitive and 14 cases of GC-insensitive. the levels of GR-beta mRNA in group of GC-insensitive [(5.72 +/- 0.58) x 10(2) copy/microg] were significantly higher than that of GC-sensitive [(4.82 +/- 0.28) x 10(2) copy/microg, t = -6.65, P < 0.01] and normal nasal mucosa [(4.44 +/- 0.35) x 10(2) copy/microg, t = -9.19, P < 0.01]. There was difference between group of GC-insensitive and other two groups of GC-sensitive and normal nasal mucosa (P < 0.01). The ratio of GR-alpha/GR-beta showed significant difference between the group of GC-sensitive (829.42 +/- 67.36) and that of GC-insensitive (535.70 +/- 89. 00, t = 11.74, P < 0.01).

CONCLUSIONSThe high expression of GR-beta mRNA and low expression of GR-alpha mRNA in nasal polyp show the status of "insensitive" in chronic sinusitis and nasal polyp. The GR-beta may play an important role in evaluating the effect of glucocorticoid in chronic sinusitis and nasal polyps.

Adult ; Case-Control Studies ; Chronic Disease ; Female ; Humans ; Male ; Nasal Mucosa ; metabolism ; Nasal Polyps ; metabolism ; RNA, Messenger ; genetics ; Receptors, Glucocorticoid ; metabolism ; Sinusitis ; metabolism

OBJECTIVETo evaluate protective effects of Rheum tanguticum polysaccharides (RTP) on traumatic brain injury (TBI) in rats.

METHODThe polysaccharides (RTP) were extracted from Tanguficum Maxim. 120 rats were divided into 15 groups, with 8 rats in each group. RTP at 100, 200 and 400 mg x kg(-1) were administrated orally once a day for five days, and model of brain injury was made by dropping weight method.

RESULTRTP reduced water content and malondialdehyde (MDA) levels, and increased total SOD activity and Na+-K+ ATPase activity after injuried.

CONCLUSIONThe polysaccharides may be one of the effective comptents in Rheum tanguticum, showing significant neuroprotective effects.

Animals ; Brain Injuries ; enzymology ; metabolism ; pathology ; Cerebral Cortex ; enzymology ; ultrastructure ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

METHODSDNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.

RESULTSOnly the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.

CONCLUSIONThe results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.

Animals ; Antigenic Variation ; COS Cells ; Cercopithecus aethiops ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Plasmids ; genetics ; Point Mutation ; Transfection

OBJECTIVETo analyze the genetic diversity of wild Rehmannia glutinosa and evaluate and compare random amplified polymorphic DNA (RAPD) and inter sample sequence repeat (ISSR) for analysis of R. glutinosa accessions.

METHODTwo molecular markers, RAPD and ISSR were used for analyzing 55 wild R. glutinosa accessions.

RESULTAverage 16.00 and 19.08 bands were amplified by RAPD primers and ISSR primers respectively, and the percentage of polymorphic bands were 89.58% and 94.32% respectively; Fifty-five R. glutinosa accessions categorized into 7 clusters were identified by unweighted pair-group method, arithmetic average (UPGMA) method.

CONCLUSIONA high level of genetic diversity of wild Rehmannia glutinosa was displayed at DNA level, and genetic diversity coefficient of R. glutinosa from different production areas was 0.63-0.98, and ISSR marker can detect higher genetic diversity of R. glutinosa germplasms than RAPD marker.

Genetic Variation ; genetics ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; methods ; Rehmannia ; classification ; genetics

Objective To investigate the effects of rapamycin on human lens epithelial cells (HLECs) against high glucose-induced oxidative stress.Methods A oxidative damage model of HLECs induced by high glucose was established,and intervention with different concentrations of rapamycin for the cells was performed for detecting the survival rate by CCK-8 assay,observing cell morphology by inverted microscope,and measuring the level of intracellular reactive oxygen species (ROS) by H2DCFDA staining,as well as determining the expression of apoptosis related Bcl-2 and Bax proteins by Western blot in all groups.Results CCK-8 results showed that the survival rate of HLECs was increased to 62.00％ ± 3.74％ and 79.57％ ± 5.26％ after treated with 10 nmol · L-1 and 100 nmol · L 1 rapamycin,and the difference was statistically significant when compared with the high glucose group (42.32％ ± 3.10％) (all P ＜ 0.05);H2DCFDA fluorescent probe staining demonstrated that rapamycin could suppress ROS generation in HLECs and alleviate oxidative damage to cells;besides,rapamycin could downregulate the expression of Bcl-2 and Bax protein in HLECs and the inhibitory effects were positively correlated with the concentration of rapamycin.Conclusion Rapamycin can inhibit high glucose-induced oxidative damage and apoptosis m human HLECs,which provides a prevention effect for diabetic cataract.

Objective To master the clinical manifestations of common cavity deformity and improve the diagnostic ability of common cavity deformity. Methods The clinical manifestations, the results of CT and MRI, and the diagnostic process of 1 patient with common cavity deformity were introduced and analyzed; related articles were compared too. Results Hearing loss from childhood was noted in the patient with common cavity deformity, and otorrhen or rhinorrhea of CSF might accompany. The patient was often misdiagnosed as having secretory otitis media, otits media suppurativa, cerebrospinal rhinorrhea or suppurative meningitis, thus, the patient might having common cavity deformity was easily ignored. Conclusion The patient with recurrently unclear cerebrospinal fluid leak and meningitis, specially accompanied hearing loss from childhood, would be suspected as having common cavity deformity. The diagnosis of the disease is based on the examination CT and MRI of the temporal bone.