OBJECTIVE:To establish a RP-HPLC method for determination of the content of baicaline in Qingkailing Injection,so as to control the quality of this preparation.METHODS:The column was C18.The mobile phase was methanol-acetic acid-water(45∶1∶55).The detection wavelength was 274nm.RESULTS:There was a good linear relationship between the concentration of baicaline and absorption area value in range of 0.25～2.5?g/ml,r=0.9 992.The average recovery was 98.4%(n=5) with RSD=1.5%(n=5).CONCLUSION:The method was highly repeatable and accurate.

Objective To study the detection and clinical signitlcanee of C reactivn protein(ClIP)and P-seleetin(P-sel)in patients with ulcerative colitis(UC).Methods The levels of peripheral blood ClIP and P-sel in patients with UC(n=53)and normal control group(n=35)were detected and the effects on disease severity were analyzed subsequently.Results The levels of blood CRP and P-sel in active phase group were significantly higher than those in catabasis group and normal controls group(P＜0.05);severe stage and medium stage were signiticandy higher than those in mild stage(P＜0.05,P＜0.01),severe stage was significantly higher than those in medium stage(P＜0.05);the blood level of CRP was positively related to P-sel in active phase group(r=0.638,P＜0.01).Conclusion CRP and P-sel were correlated with invasion and severity of UC.

Objective To study on a new generation of histone deacetylase inhibitor LBH589 on multiple myeloma (MM) resistant cells associated with changes in gene expression,and the mechanism of LBH589 reversal of MM cells' drug resistance.Methods By Western blot analysis,LBH589 (0,20,50 nmol/L) and 50 nmol/L LBH589 combined respectively with dexamethasone of 5 μmol/L at 24 h,expression of histone H4 acetylation and bcl-X gene on MMIR cells were detected.By real-time fluorescence quantitative PCR analysis,LBH589 (0,20,50 nmol/L) and 50 nmol/L LBH589 combined respectively with dexamethasone of 5 μmol/L at 24 h and 48 h,expression of TOSO on MM1R cells were detected.Results Western blot analysis showed that single LBH589 and combined with dexamethasone showed at 24 h the up-regulation on expression of the histone H4 acetylation[(0.205±0.008) ％,(0.346±0.009) ％,(0.580±0.053) ％,(0.986±0.012) ％,F =992.957,P =0.032],while down-regulation on expression of the bcl-X in MM1R cells in a dose-dependent manner[(1.210±0.160) ％,(0.930±0.036) ％,(0.730±0.017) ％,(0.488±0.029) ％,F =56.432,P =0.028].However,the group of single LBH589 was less effective than the combined group (all P ＜ 0.05).Real-time fluorescence quantitative PCR analysis showed single LBH589 and that combined with dexamethasone showed at 24 h and 48 h,the down-regulation on expression of the TOSO in MM1R cells in a dose-dependent manner,24 h specific numerical were (1.00±0.00) ％,(0.55±0.01) ％,(0.47±0.01) ％,(0.38±0.01) ％ (F =1006.344,P =0.041),48 h specific numerical were (1.00±0.00) ％,(0.39±0.04) ％,(0.05±0.01) ％,(0.03±0.00) ％ (F =2383.977,P =0.045),however,the group of single LBH589 was less effective than the combined group (all P ＜ 0.05).Conclusion Histone deacetylase inhibitor LBH589 can recover dexamethasone sensitivity of MM1R through effect the gene expression of bcl-X and TOSO,promot histone acetylation,and inducing cell apoptosis to treat tumor.

Objective To investigate the expression of DNA methylation related protein enhancer of zeste homolog 2 (EZH2) in multiple myeloma (MM) cell line U266 and enriched MM cells (MM-BMMNC) from 10 patients,and analyze the potential role of DNA methylation in the occurrence and development of MM.Methods Human MM cell line U266 and MM cells from 10 MM patients were studied.Enriched bone marrow mononuclear cells (N-BMMNC) from 10 healthy persons were used as the control.The expression of EZH2 were determined by Western blot using the ratio of target protein EZH2 and the internal reference β-actin expression as the final results.Results The preliminary in vitro results showed that EZH2 expression was higher in MM cell line U266 and patient MM-BMMNC (N-BMMNC 0.2277±0.1306,MM-BMMNC 0.9220±0.1301,U266 0.9730±0.1029),with statistically significant differences (P ＜ 0.05).Conclusion EZH2 is enriched in U266 and MM-BMMNC.DNA methylation may have a close correlation with the occurrence and development of MM.

ObjectiveTo explore the relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects (NTDs) induced by all-trans retinoic acid (RA)in Kunming mouse. Methods Fifty pregnant mice were randomly divided into control and RA-treated groups.RA-treated mice were fed with 30mg/kg RA dissolved with peanut oil on embryo 7.75 days, while the mice of control group were administrated with an equal volume of peanut oil on the same time. Then all the embryos were sampled from pregnant mice at the 4th, 18th, 42nd, 66th and 90th hour after treatment. In situ hybridization and immunohistochemical staining technique were used to detect the expression of Dishevelled2 and Vangl2 in embryonic neural tube. Results The two proteins both existed in the epithelial tissue of the mouse embryonic neural tube and displayed different expression modes at various developmental stages.Compared with the control group, the RA treated group showed a significant decrease (P≤0.05) at the 18th and 42nd hour and a significant increase (P≤0.05) at the 66th hour in Dishevelled2 protein after maternal treatment, and no significant difference was found at the 90th hour. Compared with the control group, the Vangl2 mRNA expression in the RA treated group displayed a significant decrease (P≤0.05) at the 4th and 18th hour and a significant increase (P≤0.05) at the 66th hour after RA treatment, and no difference was found at the 42nd hour. Compared with the control group, the expression of Vangl2 protein in the RA treated group decreased (P≤0.05) at the 18th and 42nd hour, and increased (P≤0.05) at the 90th hour after RA treatment, no difference was found at the 66th hour. Conclusion Excessive RA may interfere with the normal embryonic neural tube closure by regulating the expression of Dishevelled2 and Vangl2.

Objective To investigate skin manifestations and comorbidities of chronic arsenicosis due to conta?minated drinking water, and to explore their possible risk factors. Methods Data about demographic characteristics, skin manifestations and comorbidities were collected from 95 patients with chronic arsenicosis due to contaminated drinking water in Inner Mongolia, and retrospectively analyzed. A logistic regression model was established to analyze associations of skin manifestations and comorbidities with patients′ gender, age, age at onset of drinking of arsenic?contaminated water, arsenic concentrations in water and duration of arsenic exposure. Results Among the 95 patients, 77 had hyperpigmentation, 75 hypopigmentation, 93 palmoplantar keratoderma, 27 skin cancer, and 8 multiple skin cancer. Five patients were complicated by tuberculosis, 15 by hypertension, 2 by rheumatoid arthritis, 4 by cerebral infarction, 7 by coronary heart diseases, 3 by internal malignancy, 6 by hepatic cirrhosis and 2 by anemia. Logistic regression analysis revealed a significant correlation between hyperpigmentation and arsenic concentrations in water(OR=0.32, 95%CI=0.10-0.98;ORadjusted=0.27, 95%CI=0.08-0.90), between occurrence of hepatic cirrhosis and arsenic concentrations in water (OR=24.67, 95%CI=2.69-226.57;ORadjusted=22.51, 95%CI=2.38-213.11), and between occurrence of coronary heart diseases and duration of arsenic exposure(OR=6.41, 95%CI=1.09-37.88;ORadjusted=8.55, 95%CI=1.21-60.41). Conclusions There is a high incidence of aberrant pigment metabolism, palmoplantar keratoderma and skin cancer in patients with chronic arsenicosis due to contaminated drinking water. Different arsenic concentrations in water and duration of arsenic exposure seem to have different influences on the human body.

Objective To investigate the effects of Lokomat robotic-assisted gait training combined with drop foot stimulator on gait function in patients with stroke. Methods Thirty-six stroke patients were randomly divided into control group (n=18) and observation group (n=18). Both groups received routine rehabilitation training. The control group accepted Lokomat robotic-assisted gait training, while the ob-servation group worn drop foot stimulator in addition. The training intensity was 30 minutes every day, five days per week for 6 weeks. Fugl-Meyer Assessment-Lower Limb (FMA-LL), Functional Ambulation Category (FAC) and footprint analysis were used to evaluate the motor ability of lower limbs, walking ability and gait before and after training. Results There was no significant difference in the scores of FMA-LL, FAC and gait parameters (walking speed, stride width, stride length difference) before training (t<0.765, Z=0, P>0.05). The scores of FMA-LL, FAC and walking speed improved, and the stride width and stride length difference decreased after training (t>2.190, Z>3.630, P<0.05). After training, the scores of FMA-LL, FAC, walking speed and stride length difference were better in the observation group than in the control group (t>2.030, Z=-2.560, P<0.05), however, there was no significant difference in stride width between two groups (t=0.570, P>0.05). Conclusion Lokomat robotic-assisted gait training combined with foot drop stimulator could improve the lower limb motor func-tion and gait ability in patients with stroke, and the effect might be better than using Lokomat robotic-assisted gait training only.

Objective To discuss the influence of psychological education on the resourcefulness of perioperative breast cancer patients. Methods Two hundred breast cancer patients from a first-class hospital in Fujian province were randomly assigned to the control group, in which routine care was done, and an intervention group, in which psychological education curriculum was employed at three stages on the basis of routine care. The resourcefulness scale was used in both two groups before and after the intervention to evaluate the score of individual and social resourcefulness and the total score of resourcefulness in these two groups. Results As to the control group, the score on individual resourcefulness and the total score on resourcefulness before and after the intervention had no significant differences (P>0.05), but the difference in the social resourcefulness scores before and after the intervention was statistically significant (P<0.05). The differences in the scores on the individual resourcefulness, social resourcefulness and the total score on resourcefulness in the intervention group before and after the intervention were all statistically different (P<0.001). What′s more, the scores of individual resourcefulness, social resourcefulness and the total score of resourcefulness between these two groups after the intervention were statistically different (P<0.001) as well. The scores of the intervention group were significantly higher than those of the control one. Conclusion The psychological education can significantly improve the resourcefulness level of perioperative breast cancer patients, which can help individuals to manage stress effectively.

Objective To explore the effects of Sanban manipulation of spine on ankylosing spondylitis (AS) at the early-middle stage.Methods 60 AS patients at the early-middle stage were divided into drug group (n=30) and manipulation group (n=30). The drug group wastreated with sulfasalazine and the sustained-release pills of Zhengqingfengtongning, while the manipulation group were treated with Sanbanmanipulation of spine additionally. Self-developed questionnaire were used to follow up before and 1, 3 and 6 months after treatment. ResultsThe Schober index and the finger-floor distance improved more in the manipulation group than in the drug group (P<0.05) 3 months aftertreatment. All the index improved more in the manipulation group than in the drug group (P<0.05) 6 months after treatment. ConclusionSanban manipulation of spine can improve the thoracolumbar activities in the AS patients at the early-middle stage.

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine