OBJECTIVETo establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol.

METHODA LiChrospher C18 column (4.6 mm x 250 mm, 5 microm) was used. The chromatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid) and the flow rate was 1.0 mL x min.

RESULTThe linear range of laetrile was 12.87-102.94 micron x mL(-1), r = 0.999 9, paeoniflorin 24.84 - 198.7 microg x mL(-1), r = 0.9999 and paeonol 12.57-100.56 microg x mL(-1), r = 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 %.

CONCLUSIONThe method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules.

Acetophenones ; analysis ; Amygdalin ; analysis ; Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Capsules ; Chromatography, High Pressure Liquid ; methods ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glucosides ; analysis ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Polyporales ; chemistry ; Reproducibility of Results

OBJECTIVETo explore changes of B and T lymphocyte attenuator (BTLA), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAOC), reactive oxygen species (ROS), reactive nitrogen species (RNS), malondialdehyde (MDA) in ankylosing spondylitis (AS) patients, and the effect of Xinfeng Capsule (XFC) on them.

METHODSTotally 120 AS patients were assigned to two groups according to random digit table method, the XFC group (3 XFC pills each time, 3 times a day) and the SASP group (4 SASP tablets each time, twice a day), 60 in each group. All patients were treated for 3 months. Another 60 healthy subjects were recruited as a healthy control group. The expression frequency and activation levels of BTLA were detected using flow cytometry. Serum oxidative stress indices (such as SOD and CAT, TAOC, ROS, RNS, MDA) and contents of cytokines [tumor necrosis factor α (TNF-α), IL-1β, IL-4, and IL-10] were detected using enzyme-linked immunoassay (ELISA). Erythrocyte sedimentation rate (ESR) was detected using Westergren method. High-sensitivity C-reactive protein (Hs-CRP) was detected using HITACHI 7060 type automatic biochemical analyzer. Clinical efficacies of ASAS 20 and BASDAI50 were assessed using VAS. Correlation analysis between scoring for quality of life and BTLA expression frequency was performed.

RESULTS(1) Clinical efficacies of ASAS 20 and BASDAI50 were significantly better in the XFC group than in the SASP group (P < 0.01). (2) Compared with the healthy control group, BTLA expressions in the peripheral blood of AS patients decreased significantly (P <0. 05); SOD, CAT, and TAOC values significantly decreased (P < 0.01, P < 0.05); ROS, RNS, and MDA values significantly increased (P < 0.01, P < 0.05); TNF-α, IL-1β, ESR, and Hs-CRP values significantly increased (P < 0.01); IL-4 and IL-10 values decreased significantly (P < 0.01, P < 0.05). (3) Compared with pre-treatment in the same group, BTLA/CD19 + B, BTLA/CD24 + B, SOD, TAOC, IL-4, SF-36 [physical functioning (PF), social functioning (SF), role limitation due to physical problems (RP), role limitation due to emotional problems (RE), body pain (BP), mental health (MH), vitality (VT), general health (GH)] were significantly elevated; ROS, MDA, TNF-α, ESR, Hs- CRP, VAS, BASDAI and BASFI, BAS-G were significantly lower in the peripheral blood of the two groups after treatment (P < 0.01, P < 0.05). Better effect was shown in the XFC group in elevating BTLA/CD19+ B, BTLA/CD24 + B, SOD, TAOC, IL-10, BP, MH, VT, and SF; and lowering ROS, IL-1β, MDA, TNF-α, ESR, Hs-CRP, VAS, BASDAI, BASFI, and BAS-G (P < 0.01, P < 0.05). (4) Pearson correlation analysis showed, BTLA/CD19 + B expression of the peripheral blood was positively correlated with SOD, CAT, TAOC, IL-4, IL-10, GH, RP, BP, and SF (r = 0.431, 0.325, 0.318, 0.316, 0.348, 0.314, 0.358, 0.318, 0.326, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, MDA, TNF-α, IL-1β, ESR, VAS, and BASDAI (r = -0.342, -0.368, -0.334, -0.354, -0.324, -0.372, -0.342, respectively, P < 0.05, P < 0.01). BTLA/CD24 B expression of the peripheral blood was positively correlated with SOD, TAOC, IL-4, IL-10, GH, RP, BP, SF, RE, MH, VT (r = 0.358, 0.352, 0.372, 0.436, 0.435, 0.326, 0.352, 0.345, 0.326, 0.343, 0.332, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, RNS, MDA, ESR, Hs-CRP, VAS, BASDAI, and BASFI (r = -0.447, -0.336, -0.405, -0. 395, -0. 358, -0.436, -0.338, -0.425, respectively, P < 0.05, P < 0.01).

CONCLUSIONXFC could improve BTLA expression in the peripheral blood of AS patients, negatively regulate activation and proliferation of B cells, and reduce abnormal immune responses and oxidative stress injury, thereby effectively alleviating joint stiffness and pain.

B-Lymphocytes ; physiology ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Capsules ; Catalase ; metabolism ; Cytokines ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Flow Cytometry ; Humans ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-4 ; metabolism ; Malondialdehyde ; metabolism ; Oxidative Stress ; Quality of Life ; Reactive Oxygen Species ; Spondylitis, Ankylosing ; drug therapy ; Superoxide Dismutase ; metabolism ; T-Lymphocytes ; physiology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the regulation trend of Jinxin Oral Liquid (JXOL) on the expression of negative regulatory factor of TLR3 signaling pathway SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points.

METHODSTotally 75 BALB/c mice were randomly divided into 5 groups, i.e., the normal control group, the model group, the ribavirin group, the high dose JXOL group, and the equivalent dose JXOL group, 15 in each group. Each group had 3 intervention ways (I, II, and III) with 5 mice treated in each group. BALB/c mice were nasally infected with respiratory syncytial virus (RSV), and treated by different intervention ways. After intervention, mice were killed and their lung tissues were sampled, mRNA expression levels of RSV-M, SOCS1, and IFN-β were detected by Real time PCR. The expression of SOCSl at the protein level was detected by Western blot.

RESULTSCompared with the normal control group, the mRNA expression level of SOCS1 and IFN-β, and the protein expression level of SOCS1 increased significantly in the model group intervened by intervention I and II (all P < 0.01), but the mRNA expression level of IFN-β decreased significantly in model group intervened by intervention III (P < 0.01). Compared with the model group, the mRNA expression level of RSV-M all significantly decreased in the high dose JXOL group and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and III and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of IFN-β significantly decreased in the high dose JXOL group intervened by intervention I and II and the equivalent dose JXOL group intervened by intervention I (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III and the equivalent dose JXOL group intervened by intervention III (all P < 0.01). The protein expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III (all P < 0.01). Compared with the high dose JXOL group, the mRNA expression level of RSV-M decreased significantly in the equivalent dose JXOL group intervened by intervention I and II (P < 0.01). The mRNA expression level of SOCS1 and IFN-β decreased significantly in the equivalent dose JXOL group intervened by intervention I (P < 0.01), but the mRNA expression level of IFN-β increased significantly in the equivalent dose JXOL group intervened by intervention II and III (all P < 0.01). The protein expression level of SOCS1 decreased significantly in the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01).

CONCLUSIONSJXOL could inhibit the expression of SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points. Its regulatory effect might be associated with promoting the expression of interferon type I and further fighting against RSV.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; Respiratory Syncytial Virus Infections ; drug therapy ; metabolism ; Respiratory Syncytial Viruses ; Ribavirin ; Signal Transduction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism ; Toll-Like Receptor 3 ; metabolism

Quite recently,among new emerging contaminants,pharmaceutical and personal care products(PPCPs)and their active metabolites are an emerging environmental issue,due to their presence in the aquatic environment and potential for impacts on wildlife and humans.Carbamazepine is one of the most frequently and at the relatively high concentration levels detected pharmaceuticals in surface water and even in drinking water.Moreover,this drug has displayed high chronic ecotoxicity.A strain of carbamazepine-degrading bacterium was isolated from activated sludge treating pharmaceutical wastewater in Suzhou,China.It was identified as Acinetobacter sp.HY-7,based on biochemical test,16S rRNA and gyrB gene se-quence analysis.Strain HY-7 could grow in liquid mineral salt medium with carbamazepine as sole source of carbon,nitrogen and energy.HPLC analysis revealed the carbamazepine degradation percentage by HY-7 after 10 days was 48% at pH 6.0 and 25?C.Among carbamazepine and the similar structure compounds,in-dole,catechol,naphthalene,anthracene could also be utilized by strain HY-7 for growth,which exhibited a very broad substrate profile.

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA

OBJECTIVETo study the expression of alpha-catenin in the rat testis after intra-testicular testosterone withdrawal induced by injection of testosterone undecanoate (TU).

METHODSTen adult male SD rats received vehicle (n = 5 ) or TU (19 mg/kg every 15 days, n = 5) for 130 days. Paraffin-embedded testicular sections were used for immunohistochemistry against a polyclonal anti-alpha-catenin antibody.

RESULTSIn the control, alpha-catenin was expressed in the acrosome of spermatids and the cytoplasm of Leydig cells and peritubular myoid cells. In the TU-treated rat testis, Leydig cells were atrophied and the expression of alpha-catenin was markedly decreased or absent, but there was no evident change in the immunostaining of spermatids or myoid cells.

CONCLUSIONIntra-testicular testosterone withdrawal-induced looser arrangement or sloughing of spermatogenic cells is not related to the adhesion molecule alpha-catenin. Alpha-catenin may be used as a cell identification marker for Leydig cells.

Animals ; Immunohistochemistry ; Leydig Cells ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; drug effects ; metabolism ; Testosterone ; analogs & derivatives ; pharmacology ; alpha Catenin ; biosynthesis

The application potential of rep-PCR in typing beer-spoilage isolates was studied. The effects of different factors, including DNA templates and primers, on the quality and reproducibility of fingerprints were investigated. The CTAB protocol was shown to be the feasible method for DNA extraction. Primers BOXA1R and (GTG)5 were used in rep-PCR, and the PCR products were sequenced to identify strains isolated from two breweries. Rep-PCR fingerprint profiles were obtained by using GelCompar II software. Cluster analysis showed that the isolates belonging to Lactobacillus brevis, L. buchneri, L. casei/paracasei, L. plantarum are divided into 2 or 3 subgroups. In addition, the two rep-PCR fingerprint profiles complemented with each other in typing these isolates. Combining the similarity coefficient cut-off (SCC) of species, 9 unknown isolates were identified rapidly by using both fingerprint databases. The results indicate that rep-PCR is a simple, reliable and promising method for rapid identification of beer-spoilager.
Beer ; microbiology ; Cluster Analysis ; DNA Fingerprinting ; methods ; DNA, Bacterial ; genetics ; isolation & purification ; Databases, Genetic ; Lactobacillus ; genetics ; isolation & purification ; physiology ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; Time Factors

OBJECTIVETo isolate and purify components from polysaccharides of purple sweet potato (PPSP) and to test their anti-tumor activity.

METHODSDEAE-Cellulose and CM-Cellulose exchange chromatography were applied to separate components of PPSP. The anti-tumor activities of each component were measured by MTT assay on Hela and HepG(2) cells and their monosaccharide composition were analyzed by TLC chromatography, followed by infrared spectroscopy studies.

RESULTSThrough weak anion exchange chromatography and gradient elution by sodium chloride solution, four components were separated and named as PPSP, PPSPII, PPSPIII and PPSPIV, respectively. MTT tests showed that PPSP II and PPSPIII inhibited Hela and HepG2 tumor cells in a certain extent. The structural analysis revealed that PPSPI was mainly composed of glucose and galactose, PPSP II was composed of glucose and had a typical absorption peak of β-D-glucose chitosan pyranose, PPSP III was a glycoprotein showing a protein absorption peak.

CONCLUSIONFour components were separated from PPSP successfully, among which PPSP II and PPSP III shows anti-tumor activities on Hela and HepG(2) cells in vitro.

Antineoplastic Agents, Phytogenic ; pharmacology ; HeLa Cells ; Hep G2 Cells ; Humans ; Ipomoea batatas ; chemistry ; Polysaccharides ; pharmacology

OBJECTIVETo elucidate the possible mechanism of oral carcinogenesis and to explore the value of clinical application of the detection of cytokeratin (CK) 19 for oral squamous cell carcinoma (OSCC) patients.

METHODSThe cancerous tissues, para-cancerous tissues and excised lymph nodes were collected from 20 operated patients with OSCC. The patients didn't receive radiotherapy and chemotherapy before hospitalization. The relative expression of CK19 mRNA in those tissues was detected by fluorescent quantitative polymerase chain reaction (FQ-PCR).

RESULTSThe expression of CK19 mRNA in the cancerous tissues was 1.85 and 1.66 times higher than that in normal oral mucosa and in para-cancerous tissues, respectively. The expression of CK19 mRNA in lymph nodes from 9 patients with OSCC was positive and the positive rate was 45% (9/20). The positive rate of CK19 mRNA in all lymph nodes from 9 patients with OSCC was 81.8% (18/22), and the positive rate of CK19 mRNA in all lymph nodes from 20 patients with OSCC was 41.9%(18/43). CK19 mRNA level in the cancerous tissues relative to para-cancerous tissues and normal oral mucosa of the patients whose CK19 mRNA expression was positive was lower than that of the patients whose CK19 mRNA expression was negative in lymph nodes, respectively.

CONCLUSIONThe possible reason that the expression of CK19 mRNA in the cancerous tissues was higher than that in para-cancerous tissues and normal oral mucosa was that the CK19 synthesis in cancerous tissues increased obviously. The detection of CK19 mRNA in lymph nodes was regarded probably as one of the markers for detecting OSCC micrometastasis in lymph nodes. The detection of CK19 mRNA in lymph nodes by FQ-PCR was more sensitive than hematoxylin-eosin staining in diagnosing OSCC micrometastasis.

Carcinoma, Squamous Cell ; Female ; Humans ; Keratin-19 ; Male ; Middle Aged ; Mouth Mucosa ; Mouth Neoplasms ; RNA, Messenger

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.
Animals ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Cell Nucleus ; metabolism ; Cerebral Cortex ; metabolism ; Female ; Fibroblast Growth Factor 1 ; administration & dosage ; pharmacokinetics ; Gene Products, tat ; administration & dosage ; pharmacokinetics ; Hippocampus ; metabolism ; Injections, Intravenous ; Male ; Mice ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; administration & dosage ; pharmacokinetics