Objective: To investigate and characterize the O-glucuronidation pathways of the S-stereoisomer of shikonin (alkannin). Methods: Liquid chromatography and mass spectrometry were employed for the detection of akannin and its glucuronide. The incubation of alkannin in human liver microsomes (HLM), human kidney microsomes (HKM) and recombinant human UDP-glucuronyltransferases (UGT) were employed for the study of metabolism profile, the involved UGT isoforms and kinetic analysis. Recombinant human UGT screening, correlation study and chemical inhibition experiments were used for elucidation the selectivity of UGT isoform towards alkannin. Results: In the incubation of alkannin in HLM with the presence of UDPGA, a single UGT metabolite was detected. The screening of the recombinant human UGTs found that UGT1A9 high selectively catalyzed the glucuronidation of alkannin. Kinetic analysis revealed the kinetic of alkannin in HLM, HKM and recombinant UGT1A9 all followed substrate inhibition model and the Km values were 3.75-4.50 μmol/L. The glucuronidation of alkannin and propofol, a probe substrate of UGT1A9, in 12 individual HLM showed really good correlation, the correlation coefficient R2 was 0.88. Chemical inhibition experiments indicated that HLM magnolol and niflumic acid showed obvious inhibition to alkannin glucuronidation; Testerone, celastrol, and nilotinib did not inhibit alkannin glucuronidation. Conclusion: This study finds that UGT metabolism is an important metabolism pathway of alkannin in human, and alkannin is a highly selective probe substrate of human UGT1A9.

OBJECTIVETo verify the regulatory effects of acupuncture on exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) at stable phase.

METHODSThirty cases of COPD were randomly divided into a treatment group (16 cases) and a placebo group (14 cases). Based on specified aerobic exercise, acupuncture was applied in the treatment group and placebo acupuncture was used in the placebo group. The acupoints included Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and so on. The needle did not penetrate into the skin for the placebo group. The treatment was required for 2 to 3 times per week for totally 5 weeks. The indices of exercise tolerance, including 6-min walking distance (6-MWD), exercise time, maximum oxygen uptake (VO2max) forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximum ventilatory volume (MVV), St. George respiratory questionnaire (SGRQ) were observed in two groups before and after treatment.

RESULTS(1) Exercise tolerance: the differences of 6-MWD and exercise time were statistically significant between groups, which were more superior in the treatment group (both P<0.01); the VO2max was significantly increased after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05). (2) Pulmonary ventilation function: the differences of FEV1%, FEV1/FVC and MVV% were statistically significant between groups, which were more superior in the treatment group (P<0.05, P<0.01); (3) SGRQ: the SGRQ was significantly improved after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05).

CONCLUSIONThe acupuncture could improve the exercise tolerance in patients with chronic obstructive pulmonary disease at stable phase, and shorten the onset time of aerobic exercise. Besides, acupuncture combined with aerobic exercise could effectively improve the pulmonary function.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Exercise Tolerance ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

OBJECTIVETo investigate the mid-term outcome of operation for thoracolumbar tuberculosis. METHODS : Twenty-eight patiens with thoracolumbar tuberculosis underwent one stage anterior debridement,interbody fusion with bone graft and posterior pedicle screw internal fixation treatment from July 2006 to July 2011. There were 17 males and 11 females. Total 17 patients had nerve injuries ,including 6 cases of grade B, 5 cases of grade C, 6 cases of grade D according to Frankel classification. The poisoning symptoms of tuberculosis and recovery of spinal function were observed. The bone fusion and recovery of [umbar function were evaluated.

RESULTSAll the patients were followed up ,and the duration ranged from 39 to 85 months (mean 57 months). The clinical symptoms were controlled gradually, and the thoracolumbar back pain was alleviated after operation. Among the 17 patients with complications of nerve injuries, 3 patients were improved from preoperative grade B to postoperative grade D, 3 patients were improved from preoperative grade B to postopertive E, 5 patients with preoperative grade C and 6 patients with preoperative D were almostly recovered to normal after operation. According to JOA scoring system for curative effect evaluation, the excellent and good rate at the 3rd month, the 1st year, the 3rd year and the 5th year after operation were 67.86% ,82.14% ,85.71% ,89.29% and 91.30% respectively. The results at the 6th month and the 1st year had no statistical differences compared to the results at the 3rd month (P > 0.05); but the results at the 3rd year and the 5th year were better than that at 3 months after operation (P < 0.05); and the results between 3 yesrs and 5 years after operation had no statistical differences (P < O.05). The degeneration of adjacent segments were evaluated according to the California University (Universith of California at Los Angeles , UCLA) score. The degeneration rate was 53.57% (15/28) at the 3rd year after surgery, which was better than that before surgery. Twenty-three patients were followed up for 5 years ,and the degeneration rate was 86.96% (20/23) ,which was better than those of before surgery and 3 years after surgery.

CONCLUSIONThe surgical treatment for thoracolumbar spinal tuberculosis can achieve the thorough debridement, reconstruction of spinal stability, recovery of lumbar function and promote the functional recovery of the spinal cord, which is an effective method of treatment. However, the mid term follow-up showed that more severe degenerative changes were found in the postoperative adjacent segment.

Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; methods ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; physiopathology ; surgery

Objective To evaluate effects of chronic arsenic exposure and arsenic exposure time on oxidative DNA lesions in humans. Methods A cross-sectional study was conducted in 108 subjects exposed to high concentrations of arsenic in drinking water and 75 control subjects. A cohort study was conducted in 64 subjects exposed to high levels of arsenic in drinking water for 7 or 9 years. Urinary 8-oxo-7,8-dihydredeoxygnanine(8-OHdG) levels were analyzed by the enzyme-linked immunosorbent assay kit(ELISA). Urinary arsenic concentration was detected with hydride generation atomic absorption spectroscopy. Results In the cross-sectional study, the median of urinary arsenic concentration was 484.17 mg/kg Cr for the arsenic-exposed group, and 13.80 mg/kg Cr for the control group, and the difference between the two groups was statistically significant (t=32.57, P<0.01). The median of urinary 8-OHdG levels was 16.60 and 21.88 mg/kg Cr for arsenic-exposed children and adults respectively, much higher than control children(10.50 mg/kg Cr) and adults (9.11 mg/kg Cr), and the difference was statistically significant (t=5.049, 6913, all P<0.01). Urinary 8-OHdG levels were signifieandy lower for children than adults in the exposed group(t=-1.997, P<0.05). In the cohort study, the median of urinary arsenic concentration was 461.3 mg/kg Cr for the 7-year-exposed subjects and 422.90 mg/kg Cr for the 9-year-expesed subjects, and no significant difference was observed(t=-0.250, P 0.05). The median of urinary 8- OHdG levels for 9-year-exposed children and adults were 23.46 and 24.30 mg/kg Cr respectively, significantly increased compared with those of 7-year-exposed(14.29 and 18.38 mg/kg Cr), and the difference had statical signhqcanees (t= -2.949,-3.055, all P<0.01). Conclusions Chronic arsenic exposure can lead to oxidative DNA lesions in humans. The arsenic-induced DNA lesions may aggravate with the exposure time in a certain period.

OBJECTIVETo explore clinical outcomes and advantages of anterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal tuberculosis.

METHODSFrom February 2010 to February 2014, totally 82 patients with lumbar spinal tuberculosis were treated by posterior individual fixation with small-incision focus debridement,including 50 males and 32 females with an average of 50.5 years old. All patients were divided into two groups according to different procedures. Forty-nine patients in group A were treated with anterior small-incision focus debridement with posterior internal fixation through muscle spa ring at stage I ; and 33 patients in group B were treated with focus debridement with posterior internal fixation by extraperitoneal approach at stage I . Postoperative mechanical ventilation time, preoperative and postoperative Cobb angle, visual analogue scale (VAS), erythrocyte sedimentation rate (ESR) and Frankel grading were observed and compared. Postoperative complications, stability of internal fixation and bone union were compared.

RESULTSAll patients were followed-up from 15 to 36 months with an average of 23.7 months. Psoas abscess of three patients in group A and 1 patient in group B on the opposite side increased and were healed by the secondary apocenosis. The other 78 cases were healed at stage I, and no sinus tract formation, incisional hernia, leakage of cerebrospinal and occurrence of spinal tuberculosis were occurred. Fracture healing time ranged from 3 to 7 months with an average of 4.6 months. Postoperative mechanical ventilation time and VAS score in group A was better than group B. There were no statistical differences in Cobb angle, ESR and Frankel grading at the final following-up between two groups.

CONCLUSIONAnterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal according to degree of damage is a safe and effective method.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Debridement ; methods ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Treatment Outcome ; Tuberculosis, Spinal ; surgery ; Young Adult

A sensitive antibody-based lateral flow dipstick was developed for ginsenoside Re (GRe) detection. The stick consisted of a sample pad, a conjugate pad, membrane and an absorbent pad. The membrane was coated with two capture reagents, GRe-BSA conjugate and goat anti-mouse antibodies, forming a test line and a control line, respectively. The conjugate pad was saturated with colloidal gold particles coated with affinity purified monoclonal anti-GRe antibody. The visual detection limit was 200 microg x L(-1) of GRe and the reaction time was 10 min. The Panax ginseng roots were identified after these samples (10 mg) were extracted with 5 mL tap water for 30 min at room temperature, and the extracts were tested by the dipsticks and ELISA kit. The true and false P. ginseng could be distinguished with dipsticks. The dipstick could be used to detect the quality of the P. ginseng samples when the extract was diluted 100-folds. The results were compared with those obtained using an indirect competitive enzyme-linked immunosorbent assay (icELISA). The dipstick assay proved to be a sensitive and rapid tool for quality control of P. ginseng.
Animals ; Antibodies, Monoclonal ; immunology ; Counterfeit Drugs ; analysis ; Ginsenosides ; analysis ; Immunoassay ; instrumentation ; methods ; Mice ; Panax ; chemistry ; Reagent Strips ; Time Factors

OBJECTIVETo define the effects of Astragalus membranaceus on the atrial dynamics and ANP secretion in the perfused beating rabbit atria.

METHODThe experiments have been done in isolated perfused beating rabbit atria. ANP was measured by radioimmunoassay in the atrial perfusate in real-time base.

RESULTA. membranaceus (2.0, 2.5, 3.0 g L(-1)) could increase atria stroke volume from (694.70 +/- 0.01) microL g(-1) (P<0.05) to (1,003.00 +/- 8.80) microL g(-1) (P<0.001); (1,120.00 +/- 17.71) microL g(-1) and (1,195.00 +/- 8.21) microL g(-1) (P<0.001), respectively, and its could difference increase atrial pulse pressure from (0.82 +/- 0.01) kPa to (0.86 +/- 0.01) kPa (P<0.01); (0.96 +/- 0.01) kPa (P<0.001) and (1.02 +/- 0.01) kPa (P<0.001), respectively; A. membranaceus obviously increased rabbit atrial dynamics with dose-dependent manner. Simultaneously, A. membranaceus inhibited ANP secretion. Nifedipine (1.0 micromol L(-1)), a L-type Ca2+ channel inhibitor, and KB-R 7943 (10.0 micromol L(-1)), an inhibitor of reversed Na+ -Ca2+ exchanger, blocked the effects of A. membranaceus-induced augmentation of atrial dynamics but failed to modulation the inhibition of A. membranaceus on ANP secretion.

CONCLUSIONA. membranaceus increases the atrial dynamics via Na+ -Ca2+ exchanger and L-type Ca2+ channel and negatively modulates ANP secretion in beating rabbit atria.

Animals ; Astragalus membranaceus ; chemistry ; Atrial Natriuretic Factor ; metabolism ; secretion ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Heart Atria ; drug effects ; metabolism ; secretion ; Male ; Myocardial Contraction ; drug effects ; Nifedipine ; pharmacology ; Rabbits ; Radioimmunoassay ; Thiourea ; analogs & derivatives ; pharmacology

OBJECTIVETo establish in vitro culture systems of greater epithelial ridge (GER) cells from rat cochlear and to investigate the characterization, growth pattern and ultrastructure of GER cells.

METHODSUsing a combinatorial approach of enzymatic digestion and mechanical separation to allow isolation and culture of GER cells from P1 rat cochleae. The dissociated GER cells were cultured in serum-free or 10% fetal bovine serum DMEM respectively. BrdU, phalloidin, ZO1, calretinin and myosin VIIa immunostaining and scanning electron microscope observation were performed in GER cell cultures.

RESULTSThe dissociated GER cell cultures showed positive to ZO1, phalloidin and BrdU staining, but negative to myosin VIIa and calretinin. They assumed a polygonal morphology which was similar to epithelial cells and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The GER cells presented significant ability to proliferate in both conditions. Scanning electron microscope showed that there was microvillus and centre bodies but not hair cell specific stereociliary bundles on the surface of GER cultures.

CONCLUSIONSThe GER cell cultures showed significant ability to proliferate and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The dissociated GER cells expressed epithelial cell specific marker but not marker of hair cells.

Animals ; Animals, Newborn ; Cell Culture Techniques ; Cells, Cultured ; Cochlea ; cytology ; Epithelial Cells ; cytology ; Hair Cells, Auditory ; cytology ; Rats ; Rats, Sprague-Dawley

Objective To compare the analgesic effect and adverse event of butorphanol and dezocine which are combined with sufentanil and flurbiprofen axetil in PCIA,to screen out a relatively good analgesic.Methods 110 laparotomy cases from hepato-pancreato-biliary (HPB) department and 160 laparoscopy cases from general surgery(GS) department of xinqiao hospital of third medical university were included in our study.All patients were randomly divided into two groups according to the random number table method,namely the butorphanol group and dezocine group.Butorphanol 0.04 mg/kg,sufentanil 2.8 μg/kg,flurbiprofen axetil 3 mg/kg and granisetron 6 mg were used in HPB butorphanol group.Dezocine 0.2 mg/kg,sufentanil 2.8 μg/kg,flurbiprofen axetil 3 mg/kg and granisetron 6 mg were used in HPB dezocine group.Butorphanol 0.04 mg/kg sufentanil 2.5 μg/kg,flurbiprofen axetil 3 mg/kg and granisetron 6 mg was used in GS butorphanol group.Dezocine 0.2 mg/kg,sufentanil 2.5 μg/kg,flurbiprofen axetil 3 mg/kg and granisetron 6 mg was used in GS dezocine group.The mean arterial pressure(MAP),heart rate(HR),facial expressions of pain score,sedation score,PONV score,NRS score and respiratory depression were observed in postoperative 0 hour,6 hours,24 hours,48 hours.Results For both two departments,the numbers of patients with NRS score and facial expressions of pain scores greater than 3 in dezocine group were more than those in butorphanol group,the differences were significant(P ＜ 0.05).There was no statistically significant difference in numbers of patients with NRS score and facial expressions scale of 1 to 3 (P ＞ 0.05).while the number of cases with sedation score ranged from 1 to 3 in dezocine group was less than that in both HPB and GS butorphanol group(P ＜ 0.05).There was no statistically significant difference in PONV score and itching score which was or less than 3 or more than 3 (P ＞ 0.05).Conclusion For postoperative analgesia in PCIA,butorphanol has better analgesic effect than the same dose of dezocine,and stronger sedation effect than dezocine.

Objective: To study the mechanism of chitosan i n inhibiting the proliferation of rabbit aortic smooth muscle cells(SMCs). Methods: By means of c-myc probe labelled with random primers and Northern blot hybridization, we examined the effect of chitosan on vascu lar SMC c- myc mRNA expression, which was stimulated by newborn bull serum (NB S,20%). Results: The oncogene c-myc mRNA expression incerased in cultured vascular SMC 24 h after NBS exposure. These effects were inhibite d by chitosan (20 μg/ml). Conclusion: Chitosan might inhibit the expression of vascular SMC c-myc mRNA stimulated by NBS, through which the proliferation of vascular SMC are inhibited.