OBJECTIVETo observe the therapeutic effect and relevant mechanism of shuxuening Injection (SI) in treating patients with active ulcerative colitis (UC).

METHODSTotally 91 patients with active UC were randomly assigned to 2 groups, 44 in the control group and 47 in the treatment group. Patients in the control group received routine treatment, while patients in the treatment group additionally received intravenous injection of SI (15 mL), twice daily for 14 days in total. Colonoscopy was performed before and after treatment. The therapeutic effect was assessed by Mayo scoring system and the grading of activities evaluated by Baron endoscope. Serum levels of IL-6 and TNF-α were detected by ELISA. The activity of SOD was detected by xanthine oxidase method. The content of MDA was detected by thiobarbituricacid (TBA). Besides, 20 healthy subjects were recruited as the healthy control group.

RESULTSTotally 82 patients completed the study (40 in the control group and 42 in the treatment group). There was no statistical difference in serum levels of IL-6, TNF-α, SOD, MDA, the Mayo score and endoscope grading between the two groups before treatment (P >0. 05). Compared with the healthy control group, serum levels of IL-6, TNF-α, MDA significantly increased (P <0.01), and the serum SOD level decreased (P < 0. 05) in the treatment grup and the control group before treatment. Compared with before treatment in the same group, serum levels of IL-6, TNF-α, MDA, the Mayo score and endoscope grading all decreased in the treatment group and the control group after treatment (P <0. 01, P <0. 05). Compared with the control group after treatment, serum levels of IL-6, TNF-α, MDA, the Mayo score and endoscope grading all decreased (P <0.01, P <0.05), the serum SOD level increased (P <0.05) in the treatment group after treatment. The serum SOD level was obviously negative correlated with serum levels of IL-6, TNF-a, Mayo score, and endoscope score (r = -0. 621, -0.638, -0. 509, -0.787, P <0.01). The serum MDA level was obviously positive correlated with serum levels of IL-6, TNF-α, Mayo score, and endoscope score (r =0.711, 0. 882, 0. 525, 0. 639, P <0.01).

CONCLUSIONSI could improve inflammatory injury and clinical symptoms of patients with active UC, and its mechanism might be associated with antioxidant and scavenging oxygen free radicals.

Colitis, Ulcerative ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Inflammation ; drug therapy ; Interleukin-6 ; blood ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo understand the possibility of estimating the anti-inflammation effect of Liuwei Dihuang Wan (LWW) in rat by having developed HPLC-UV metabonomic technology.

METHODThe hydrophilic or lipophilic constituent group of LWW was extracted by distilled water or acetic ether respectively. The anti-inflammation effects of different LWW dosages and extractions were measured by traditional method respectively. Then, metabonomic analysis was performed.

RESULTThe high dosage of LWW extraction (16.5 g x kg(-1))could inhibit the swell of rat palm significantly, but there are not statistically significant effect of low dosage group. Nevertheless, the metabonomic study showed that LWW extraction could restore obviously the rat HPLC-UV urinary profiling disturbed by inflammation in low dosage, especially the hydrophilic constituent group.

CONCLUSIONOur study indicated that the developed metabonomic technology based on HPLC-UV might be used as a potentially powerful tool for estimating the anti-inflammation effect of LWW with sensitivity and integrity.

Animals ; Anti-Inflammatory Agents ; analysis ; metabolism ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; metabolism ; pharmacology ; Male ; Metabolomics ; methods ; Rats ; Rats, Sprague-Dawley

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

OBJECTIVETo explore the protection of high intensity microwave radiation on hypothalamo-pituitary-adrenal axis (HPAA) activity and hippocampal CA1 structure in rats and the protectiveeffect of Qindan Granule (QG) on radiation injured rats.

METHODSTotally 48 Wistar rats were randomlydivided into 8 groups, i.e., the normal control group, post-radiation day 1, 7, and 10 groups, 7 and 10days prevention groups, day 7 and 10 treatment groups, 6 in each group. Rats in prevention groups wererespectively administered with QG liquid (1 mL/100 g, 4. 75 g crude drugs) for 7 days and 10 days bygastrogavage and then microwave radiation. Then preventive effect for radiation injury was statisticallycalculated with the normal control group and the post-radiation day 1 group. Rats in treatment groupswere firstly irradiated, and then administered with QG liquid (1 mL/100 g, 4.75 g crude drugs). Finally preventive effect for radiation injury was statistically calculated with the normal control group, post-radiation day 7 and 10 groups. Contents of corticotrophin releasing hormone (CRH), beta endorphin (beta-EP), adrenocorticotropic hormone (ACTH), and heat shock protein 70 (HSP70) were detected. Morphological changes and structure of hippocampal CA1 region were observed under light microscope.

RESULTSCompared with the normal control group, contents of CRH and beta-EP significantly decreased in each radiation group. Serum contents of ACTH and beta-EP significantly increased in post-radiation day 1 and 7 groups (P < 0.05). Compared with radiation groups, beta-EP content in serum and pituitary significantly increased, and serum ACTH content significantly decreased in prevention groups (P < 0.05). Pituitary contents of CRH and beta-EP significantly increased in prevention groups. Serum contents of ACTH, beta-EP, and HSP70 were significantly lower in day 7 treatment group than post-radiation day 7 group (P < 0.05). Morphological results showed that pyramidal neurons in the hippocampal CA1 region arranged in disorder, with swollen cells, shrunken and condensed nucleus, dark dyeing cytoplasm, unclear structure. Vessels in partial regions were dilated with static blood; tissues were swollen and sparse. In prevention and treatment groups pathological damage of hippocampal CA1 region was obviously attenuated; neurons were arranged more regularly; swollen, pycnotic, or deleted neuron number were decreased; vascular dilatation and congestion was lessened.

CONCLUSIONQG could affect HPAA function and activity of high intensity microwave radiated rats, showing certain preventive and therapeutic effects of microwave radiated rats by adjusting synthesis and release of partial bioactive peptides and hormones in HPAA, improving pathological injury in hippocampal CA1 region.

Adrenocorticotropic Hormone ; blood ; Animals ; CA1 Region, Hippocampal ; drug effects ; pathology ; radiation effects ; Corticotropin-Releasing Hormone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; HSP70 Heat-Shock Proteins ; blood ; Hypothalamo-Hypophyseal System ; drug effects ; radiation effects ; Microwaves ; adverse effects ; Pituitary-Adrenal System ; drug effects ; radiation effects ; Random Allocation ; Rats ; Rats, Wistar ; beta-Endorphin ; blood ; metabolism

Using the qualified rates of particles as the evaluation indexes, the impact tactors of one-step pelletization technology of Jiuwei Xifeng granules were selected from six factors by the Plackett-Burman experimental design and the levels of non-significant factors were identified. According to the Plackett-Burman experimental design, choosing the qualified rates of particles and angle of repose as the evaluation indexes, three levels of the three factors were selected by Box-Behnken of central composite design to optimize the experimental. The best conditions were as follows: the fluid extract was sprayed with frequency of 29 r . min-1, inlet air temperature was 90 °C, the frequency of fan was 34 Hz. Under the response surface methodology optimized scheme, the average experimental results are similar to the predicted values, and surface methodology could be used in the optimization of one-step pelletization for Chinese materia medica.
Air Movements ; Analysis of Variance ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Hot Temperature ; Models, Theoretical ; Research Design ; Tablets

AIM: To investigate the changes and effects of orthokeratology on corneal morphology. METHODS: Totally 90 patients were treated with orthokeratology from January 2015 to December 2016. They were divided into observation group (overnight wearing) and control group(daytime wearing),45 cases (90 eyes) in each. The central corneal thickness, corneal curvature, spherical equivalent (SE), uncorrected visual acuity (UCVA) of both groups were compared before and after wearing orthokeratology lens for 1wk,1,3,and 6mo. RESULTS: The central corneal thickness of two groups before wearing glasses was significantly higher than that of the cornea after wearing glasses 1wk,1,3 and 6mo(all P<0.05); the central corneal thickness of the observation group at 1, 3 and 6mo after wearing glasses was significantly lower than that of the control group (P<0 05). The corneal curvature values of the two groups before wearing glasses were significantly higher than that of the cornea after wearing for 1wk,1,3 and 6mo (all P<0.05). The corneal curvature of observation group at 6mo was significantly lower than that of the control group (40.0士0.5D vs 41.3士0.9D, P<0.05). The staining rate of corneal epithelium was observed at 3mo after wearing glasses. The positive rate of epithelial staining was 49% (44/90) in the observation group and 29% (26/90) in the comparison group;the grade 0,grade 1 accounted for the majority of the two groups. With the orthokeratology lens wearing longer, the SE level of two groups showed a downward trend. The spherical equivalent of observation group at 6mo was significantly lower than that of the control group (-0.42士0.20D vs -0 52士0.19D, P<0.05). The UCVA value of two groups after wearing glasses significantly increased than that before wearing glasses (all P<0.05). CONCLUSION: Wearing orthokeratology lens can reduce myopia degree. Wearing it overnight has the better outcome than wearing in the daytime.

OBJECTIVESTo establish a primary biliary cirrhosis (PBC) model by AMAM2 autoantigen injection into C57BL/6 mice.

METHODSMice of the model group were immunized intraperitonealy with 200 microl of purified recombinant AMAM2 autoantigen in complete Freund's adjuvant (CFA). Mice immunized with bovine serum albumin and CFA in the same way were used as negative controls. Sixty-six weeks later, mice were sacrificed and their sera were collected. Sera samples were assayed for AMAM2 autoantibody, alkaline phosphatase (ALP), ALT and total bilirubin (TBil). Their liver, stomach, muscle and kidney tissues were sectioned and stained using HE to observe the pathological changes.

RESULTSAntibodies to AMAM2 autoantigen were readily induced in the model group. The mice in the model group had no significant changes in the level of serum ALT and TBil but had an obvious increase of ALP (P<0.05). The stomach, muscle and kidney tissues showed no evident damage while the livers had obvious pathological changes, including bile duct degeneration or proliferation, and mononuclear cell infiltration.

CONCLUSIONThe AMAM2 autoantigen-induced PBC animal model was successfully established in C57BL/6 mice in our experiment and its characteristic biochemical and pathology are quite similar to that in the early stage of human PBC. This model may provide a useful experimental approach for further study of the pathogenesis and clinical treatment of human PBC.

Animals ; Autoantigens ; immunology ; Disease Models, Animal ; Liver Cirrhosis, Biliary ; etiology ; Mice ; Mice, Inbred C57BL ; Mitochondria ; immunology

OBJECTIVETo investigate the biochemical metabolic changes detected by phosphorus-31 MR spectroscopy ((31)P MRS) with pathologic changes in the liver of fasting rabbits.

METHODSA total of 22 rabbits were under the starvation up to death to establish animal models. Hepatic (31)P MRS was performed in different period of 10 rabbits including normal condition, over-starvation, agonal condition and death after 30 min. Other 9 rabbits were divided into three type including over-starvation, agonal condition and death group with 3 rabbits in each group, and 3 healthy rabbits served as controls. All the 12 rabbits were sacrificed for the hepatic pathological examination. The MR examination was performed on a 1.5 T imager using a 1H/31P surface coil by the 2D chemical shift imaging technique. The relative quantities of phosphomonoesters (PME), phosphodiesters (PDE), inorganic phosphate (Pi) and adenosine triphosphate (ATP) were measured.

RESULTSAll the relative quantification of phosphorus metabolites were changed significantly from starvation to death (X(2)=23.13-35.41, P<0.01). The relative quantifications of ATP of normal condition, over-starvation, agonal condition and death were 2.54 +/-0.53, 1.73 +/-0.14, 0.88 +/-0.23 and 0.05 +/-0.08, respectively (rs=1.0, P<0.01). The relative quantifications of PDE from normal to death were 1.25 +/-0.54, 2.76 +/-0.23, 3.33 +/-0.49 and 3.87 +/-0.43, respectively, and those of Pi were 0.42 +/-0.02, 0.65 +/-0.05, 0.89 +/-0.15 and 0.99 +/-0.08, respectively (rs=1.0, P <0.01). The relative quantifications of PME were also significantly changed (rs=0.4, P=0.6). The pathologic changes of normal condition, over-starvation, agonal condition and death: decreased size of hepatocytes, loss of cell number, cellular swelling, degeneration and cell necrosis or hepatic hemorrhage became more and more pronounced.

CONCLUSION(31)P MRS can monitor dynamic changes of relative quantification of phosphorus metabolites, which are correlated with the pathological severity of acute hepatic injury by fasting.

Animals ; Death ; Dose-Response Relationship, Radiation ; Female ; Liver ; metabolism ; pathology ; Magnetic Resonance Spectroscopy ; methods ; Male ; Phosphorus ; metabolism ; Phosphorus Isotopes ; metabolism ; Rabbits ; Random Allocation ; Starvation

OBJECTIVETo construct luxS mutant aften luxS gene of Streptococcus mutans (S. mutans) was knocked out, and examine their ability of biofilm formation.

METHODSA recombinant plasmid containing the flanking fragment of luxS of S. mutans was transformed into S. mutans UA159, and selected by brain heart infusion (BHI) agar medium with kanamicin. The luxS mutant further confirmed via polymerase chain reaction (PCR) and the autoinducer-2 (AI-2) bioluminescence assay of Vibrio harveyi (V. harveyi), and ability of luxS mutant and S. mutans UA159 biofilm formation was examined in different phases, in BHI medium with 1% sucrose and 1% glycose by scanning electron microscopy (SEM).

RESULTSLuxS-deficient S. mutans strains were successfully constructed. Compared with S. mutans UA159, the luxS mutant maintained in BHI medium containing 1% sucrose displayed an apparent defect in biofilm formation, while they showed no significant deviation in BHI medium containing 1% glycose.

CONCLUSIONluxS gene in S.mutans can play a role in dental plaque biofilm formation, and the luxS gene is possible to regulate sucrose-dependent biofilm formation.

Bacterial Proteins ; Biofilms ; Carbon-Sulfur Lyases ; Culture Media ; Dental Plaque ; Homoserine ; analogs & derivatives ; Humans ; Lactones ; Streptococcus mutans

The metabolism, distribution and excretion profiles of recombinant human thrombopoietin (rhTPO) in mice were studied by means of (125)I-labeled rhTPO ((125)I-rhTPO) combined with size exclusive high performance liquid chromatography (SHPLC) or trichloroacetic acid (TCA) precipitation analysis. (125)I-rhTPO was prepared by iodogen method. Purification was performed on Sephacryl S-200 HR gel. Radioactive-purity of (125)I-rhTPO identified by SHPLC was (96.9 +/- 1.5)% (n = 3). The proliferation effect of TPO dependent cell line (TD-3) and the increase of peripheral platelet counts in mouse by (125)I-rhTPO demonstrated that (125)I-labeled protein maintained the biological activities of TPO both in vitro and in vivo. SHPLC analysis of serum and urine samples taken after sc 1 micro g/mouse (345 kBq/mouse) of (125)I-rhTPO revealed that there were two lower molecular weight (125)I-degradation metabolites ((125)I-MI and (125)I-MII) other than parent molecule. (125)I-MI was mainly found in urine, and (125)I-MII was detected both in serum and in urine. The maximal concentration of (125)I-rhTPO was reached at 2 hours after injection. The terminal half-life was 10.8 hours, which was much longer than those of other peptides. TCA precipitable radioactivity in tissue showed that the radioactivity in bone marrow was rather high. The highest level was found in urinary system. Levels in adrenals, lymph nodes, and fat were near to that in serum. Lowest was found in brain. The main excretion route was urinary system and (98 +/- 5.6)% of (125)I-rhTPO was excreted within 72 hours after dosing.