OBJECTIVETo observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.

RESULTSRP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).

CONCLUSIONSHJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Animals ; Berberine ; Berberine Alkaloids ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavanones ; Flavonoids ; Glucose ; metabolism ; Iridoids ; Models, Animal ; Neurons ; Oxygen ; metabolism ; Rats ; Reperfusion Injury ; drug therapy

Objective To evaluate the effectiveness of standardized three-stage rehabilitation program on spasticity and motor function in the upper extremities after cerebral hemorrhage.Methods A total of 364 patients were included and randomly assigned to a control group (n =181) and a rehabilitation group (n =183).The standardized three-stage rehabilitation program,which included early-stage bedside rehabilitation,specialized treatment in rehabilitation ward during recovery and rehabilitation follow-up at regular intervals was applied in the rehabilitation group,but only rehabilitation guidance and follow-up after discharge were provided for the control group.The modified Ashworth scale (MAS) and Fugl-Meyer assessment (FMA) were performed at the time of recruitment,1 month (M1),3 months(M3) and 6 months(M6) later.Results There was no statistical difference between the groups at recruitment.The occurrence rate of spasticity was 22.7％ in the control and 23.5％ in the rehabilitation group.At M6 the occurrence rate of spasticity was about 59.7％ and 43.2％ in control group and rehabilitation group respectively,and the number of patients grade 1 + and grade 2 on the MAS was 50/181 in the control group,significantly more than in the rehabilitation group (25/183).At all time points,MAS grade 0 accounted for a large proportion of both groups.At M6,both MAS distributions and scores of the two groups were different statistically (P ＜ 0.01).FMA scores in both groups increased significantly (P ＜ 0.01) with time,with the score being (17.13 ± 16.46),(24.87±18.36),(30.68±19.41) at M1,M3 and M6 in the control group and (24.71 ±19.80),(39.83 ± 19.50),(48.87 ± 18.25) in the rehabilitation group,but the average scores of the latter were consistently significantly higher than the former (P ＜ 0.01).Conclusions Standardized three-stage rehabilitation can alleviate spasticity and improve motor function of the upper extremities in cerebral hemorrhage patients.

AIM: To study the effect of extract from Pongamia pinnata roots on anti-inflammation and analgesia and acute toxicity. METHODS: The models of mice ear edema induced by xylene and Cotton pellet granuloma in rats to observe the anti-inflammation effect of PRE via oral administration. The effect of PRE on analgesia was tested by measuring the latent period licking hind foot with the hot plate method and counting body twisting induced by acetic acid in mice. The acute toxicity of PRE was measured by the method of Bliss. RESULTS: PRE could significantly inhibit the ear edema caused by xylene in mice, granuloma hyperplasia caused by cotton in rats. It could significantly prolong the pain threshold on hot-plate in mice, reduce the writhing times in mice. The LD50 of PRE was 6. 371 8 g/kg, its 95% confident limit was 5. 408 4-7. 723 2 g/kg. CONCLUSION: PRE has obvious effect on anti-inflammation and analgesia and the lower acute toxicity.

OBJECTIVETo optimize the synthetic pathway and fermentation process of yeast cell factories for production of oleanoic acid.

METHODUsing the DNA assembler method, one copy of Glycyrrhiza glabra beta-amyrin synthase (GgbAS), Medicago truncatula oleanolic acid synthase (MtOAS) and Arabidopsis thaliana cytochrome P450 reductase 1 (AtCPR1) genes were introduced into Saccharomyces cerevisiae strain BY-OA, resulting in strain BY-20A. YPD medium with different glucose concentration were then used to cultivate strain BY-2OA.

RESULTIncreasing gene copies of GgbAS, MtOAS and AtCPR1 resulted in increased beta-amyrin and oleanolic acid production. The strain BY-2OA produced 136.5 mg x L(-1) beta-amyrin and 92.5 mg x L(-1) oleanolic acid, which were 54% and 30% higher than the parent strain BY-OA. Finally, the titer of oleanolic acid increased to 165.7 mg x L(-1) when cultivated in YPD medium with 40 mg x L(-1) glucose.

CONCLUSIONProduction of oleanoic acid increased significantly in the yeast strain BY-2OA, which can provide the basis for creating an alternative way for production of oleanoic acid in place of extraction from plant sources.

Biomass ; Biotechnology ; methods ; Dose-Response Relationship, Drug ; Fermentation ; Glucose ; pharmacology ; Oleanolic Acid ; biosynthesis ; Saccharomyces cerevisiae ; cytology ; drug effects ; metabolism

The purpose of this study was to investigate the molecular mechanisms whereby hydrogen sulfide (H2S) exerts the promoting effect on vascular endothelial cells migration. We used wound healing assay to study the effect of NaHS (H2S donor) on the migration ability of rhesus retinal pigment epithelial cell line, RF/6A cells, under normoxic conditions. Real-time PCR was used to measure hypoxia-inducible factor 1α (HIF-1α) mRNA level. Western blot was used to measure the expression of HIF-1α protein. The probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) was used to measure intracellular reactive oxygen species (ROS) level. The results showed that NaHS (10-100 μmol/L) could significantly promote RF/6A cells migration under normoxic conditions, and this effect could be inhibited by 50 µmol/L HIF-1 inhibitor, CdCl2. NaHS increased the protein level of HIF-1α in a dose- and time-dependent manner, and up-regulated the mRNA level of HIF-1α quickly and continuously. Moreover, NaHS could significantly decrease ROS levels in RF/6A cells under normoxic conditions. These results suggest HIF-1 may mediate the promoting effect of H2S on vascular endothelial cells migration under normoxic conditions. ROS, as an upstream regulator of HIF-1α, may be involved in the migration-promoting effect of H2S.
Animals ; Cell Line ; Cell Movement ; physiology ; Endothelial Cells ; cytology ; Hydrogen Sulfide ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Macaca mulatta ; RNA, Messenger ; genetics ; metabolism ; Reactive Oxygen Species ; metabolism ; Retinal Pigment Epithelium ; cytology ; Sulfides ; pharmacology

OBJECTIVETo screen and characterize the short peptides which bind specifically to interleukin-2 (IL-2) receptor alpha chain (IL-2Ralpha) for acquisition of small antagonists for blocking the binding of IL-2 with IL-2Ralpha.

METHODS12-mer phage displayed peptide library was screened with the target cells of MT-2 cells which expressed IL-2Ralpha at high levels. The binding phage clones were eluted by anti-IL-2Ralpha monoclonal antibody. After 3 rounds of screening, the positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, and the amino acid sequences of the positive clones were deduced from the DNA sequences.

RESULTSSeven positive clones were screened out of the 17 phage clones bound to MT-2 cells. The positive clone M15 could bind specifically to MT-2 cell and PHA-activated peripheral blood monouclear cells. Amino acid sequence analysis identified 6 sequences, all of which contained hydrophilic residues, and 5 of these 6 sequences included Tyr, Phe and Leu conservative residues.

CONCLUSIONThe peptide sequences containing Tyr, Phe conservative residues identified in this study can bind to cell surface IL-2Ralpha.

Amino Acid Sequence ; Cell Line, Transformed ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Peptide Library ; Peptides ; genetics ; metabolism ; Protein Binding ; Receptors, Cell Surface ; metabolism ; T-Lymphocytes ; cytology ; metabolism

OBJECTIVETo observe the efficacy differences between electroacupuncture and physiotherapy on the proprioception of athletes with functional ankle instability (FAD.

METHODSFifty athletes with FAI were randomly divided into an electroacupuncture group and a physiotherapy group. The electroacupuncture group was treated with electroacupuncture at Jiexi (ST 41), Kunlun (BL 60), Qiuxu (GB 40) and Ashi acupoints, and the physiotherapy group was treated with low frequency electrical stimulation and infrared radiation at medial malleolus and lateral malleolus, thrice each week for consecutive 8 weeks. The Joint Position Sense: Active (JPSA), Joint Position Sense: Passive (JPSP) and Kinaesthesia (KT) were assessed at the ankle by use of Biodex System-III isokinetic dynamometer.

RESULTSThe JPSA of 11.090 +/- 3.1 degrees and the JPSP of 9.67 degrees +/- 2.8 degrees before the treatment reduced to 9.14 degrees +/- 4.0 degrees and 6.89 degrees +/- 3.3 degrees, respectively, after the treatment in the electroacupuncture group, with significant differences in JPSA and JPSP (both P < 0.05), compared with those in the physiotherary group, there were significant differences (both P < 0.05) and with no significant difference in KT (P > 0.05). There was no significant differences in the indices of JPSA, JPSP and KT in the physiotherapy group after 8 weeks than those before treatment (all P > 0.05).

CONCLUSIONElectroacupuncture can effectively improve the proprioception of athletes with FAI and achieves a superior efficacy as compared with the conventional physiotherapy.

Adult ; Ankle Injuries ; physiopathology ; therapy ; Ankle Joint ; physiopathology ; Athletes ; Electroacupuncture ; Female ; Humans ; Joint Instability ; physiopathology ; therapy ; Male ; Proprioception ; Young Adult

OBJECTIVETo prepare monoclonal antibody (mAb) against prM epitope.

METHODSThe gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.

RESULTSmAb against prM epitope of JEV was prepared successfully.

CONCLUSIONThe obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; isolation & purification ; Antibody Specificity ; BALB 3T3 Cells ; Cell Line ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Encephalitis Virus, Japanese ; genetics ; immunology ; Epitopes ; immunology ; Escherichia coli ; genetics ; Mice ; Plasmids ; genetics ; metabolism ; Prokaryotic Cells ; metabolism ; Sequence Analysis, DNA ; Viral Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification

Increased vascular endothelial growth factor (VEGF) biosynthesis in vascular endothelial cells has been reported to play an obligatory role in promoting angiogenesis. Nevertheless, the intracellular signaling mechanisms of hypoxia-induced VEGF release remain largely unknown. Human umbilical vein endothelial cell lines (ECV304) were cultured in normoxic or hypoxic conditions for 12 approximately 24 h and harvested for determination of VEGF mRNA expression and phosphorylation of ERK1/2 and p38 mitogen-activated protein kinase (p38 MAPK) by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Secreted VEGF protein was measured by enzyme-linked immunosorbent assay (ELISA). It has reported that PD98059, an ERK inhibitor, was able to blunt the hypoxia-induced activation of the expression of VEGF gene. In accordance with this report, an increase in ERK1/2 phosphorylation and VEGF biosynthesis was observed in ECV304 cells cultured in hypoxia, and this increase was blocked by PD98059. The novel finding of the present study is that an activation of p38 MAPK is involved in hypoxia-induced increase in VEGF biosynthesis. SB202190, an inhibitor of p38 MAPK was able to blunt the hypoxia-induced increase in VEGF biosynthesis. These dada provide the first direct evidence for a role of p38 MAPK in mediating hypoxia-induced increase in VEGF biosynthesis in human endothelial cells.
Cell Hypoxia ; Cell Line ; Endothelial Cells ; cytology ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; RNA, Messenger ; genetics ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology

A two-stage pH control method was employed in batch fermentation of curdlan by Alcaligenes faecalis WX-C12 where cell-growth stage was constantly controlled at pH 7.0 and stationary stage was controlled at a constant pH as well. The influence of pH control on the curdlan production was investigated. The optimal pH control of batch process for curdlan production was obtained when cell-growth stage was controlled at pH 7.0 and stationary stage was constantly controlled at pH 5.6. Production and productivity of curdlan, QP and YP/S reached 28.19 g/L, 291 mg/(L.h), 132.27 mg/(L.h.g) and 0.659, an improvement of 20.4%, 38.1%, 38.1% and 29.5% compared to a pH uncontrolled operation respectively.
Alcaligenes ; growth & development ; metabolism ; Fermentation ; Glucans ; biosynthesis ; Hydrogen-Ion Concentration ; beta-Glucans