Metabolism of n-alkane mainly includes ?-oxidation pathway and ?-oxidation pathway in Candida tropicalis,which are both catalyzed by a series of enzymes. The transformation from n-alkane to fatty acid is accomplished via ?-oxidation pathway,and the degradation of fatty acid is involved in ?-oxidation pathway. The mechanism on metabolism of n-alkane and studies on related enzymes are summarized in the review. Furthermore,some molecular genetics and metabolic engineering techniques related to Candida tropicalis developed in recent years are introduced.

Acyl Coenzyme A Oxidase(ACO) is the rate-limiting enzyme of the ?-oxidation of dicarboxylic acid in Candida tropicalis . Using strains with low Acyl Coenzyme A Oxidase activity to produce dicarboxylic acid on an industrial scale will facilitates the improvement of the yield and purity of dicarboxylic acid. In this research, we improved the method of Allain, so as to determine the activity of Acyl Coenzyme A Oxidase from Candida tropicalis easily and correctly. This method can be used as quantity criterion in the screen of industrial strains of dicarboxylic acid production, and this work established the basic of character study of Acyl Coenzyme A Oxidase. We determined the activities of Acyl Coenzyme A Oxidase from a series of Candida tropicalis strains with different ability of dicarboxylic acid production, and found the obvious relationship of ACO activity and ability of acid production.

BACKGROUND: Chronic musculoskeletal pain can cause disability and decreased quality of life of patients. Probing into the therapeutic methods of musculoskeletal disease and choosing proper treatment drugs are important for patients.OBJECTIVE: To evaluate the efficacy and tolerance of controlled-release tramadol in patients with moderate to severe chronic pain of musculoskeletal origin.DESIGN: A self-control study was conducted.SETTING: Arthrtosis Institute, People' s Hospital Affiliated to Peking University.PARTICIPANTS: Patients who were in the Orthopaedic Outpatient Service,People' s Hospital of Peking University were involved. This observational study was conducted in 40 adults who experienced moderate to moderately severe chronic musculoskeletal pain that was not controlled by routine nonsteroidal anti-inflammatory drugs(NSAID) therapy.INTERVENTIONS: Any NSAIDs and other analgesics were prohibited. Patients initially took 50 mg of controlled-release tramadol (Tramcontin) every 12hours, supplemented by every 50 mg if insufficient pain relieved. Recommended daily maximum dose was 400 mg per day. The observation lasted for 4 weeks. The patients were asked for document therapy-relevant data every day and were interviewed every week to doctor assessment.MAIN OUTCOME MEASURES: Pain intensity before and after treatment, pain remission degree, all side effects and their severity.RESULTS: All cases were finished the study and no one withdrew from follow-up. No eligible patients exited from the study because of therapeutic failure or intolerance. In comparison with the initial visual analogue scale (VAS) mean value 6. 80 ± 1.84, the average VAS value at the end of observation was 1.00 ± 1.46, which had significant difference( P ＜ 0. 001) . An average pain reduction of (85. 50±5.35)%was obtained, among which 78% were absolutely and obviously lessened. Side effects were reported in 9cases (22%).Drowsiness and dizziness(6 cases) was the most common, followed by nausea (3 cases), vomiting (1 case), somnolence (2 cases), constipation and urinary retention (1 case) and blurred vision (1 case).CONCLUSION: Tramcontin, as slow-release tramadol, proved to be an effective, safe, and easy-to-use central acting analgesic has an important role in the management of chronic musculoskeletal pain. Light side effect and good tolerance has provided a new choice for treatment of chronic pain in the orthopaedic department.

Calcinosis ; complications ; Decalcification Technique ; Humans ; Meningeal Neoplasms ; complications ; Meningioma ; complications ; Specimen Handling ; methods ; Ultrasonics

Objective To explore the value of color Doppler ultrasound detection renal interlobar arterial hemodynamic parameters for early diagnosis in patients with chronic kidney disease(CKD).MethodsSeventy-eight patients with CKD were undergone interlobar arterial hemodynamic parameters detection and 65 normal persons served as the control.The peak systolic velocity (PSV),end-diastolic velocity (EDV),pulsatility index(PI),resistive index (RI) of interlobar arterial and serum cystatin C (SCysC) were measured.The variation of RI and the correlation of renal interlobar arterial RI and pathological findings in early CKD was analyzed.Results In 40 CKD cases with normal SCysC(group A),the PSV,EDV,PI,RI of renal interlobar arterial were not statistically significant (P＞0.05).In 38 CKD cases with increased SCysC(group B),the RI of renal interlobar arterial were higher than those of group A and control group (P＜0.05),and the PSV and EDV were lower than those of group A and control group (P＜0.05).In cases of CKD with normal RI,the disease limited to the glomeruli;while the RI increased,the disease locate in the tubulointerstitial compartment.Conclusions Interlobar arterial RI united SCysC is a sensitive indicator in assessing early injury of chronic kidney disease.

Parkinson's disease (PD) is a degenerative disease of the central nervous system due to the loss or death of dopaminergic neurons in the substantia nigra. Clinically, levodopa is the most effective and commonly used drug for PD treatment. However, long-term levodopa therapy is prone to motor complications and other side effects caused by excessive peripheral dopamine production, which has become an urgent problem to be solved in PD treatment. Dopamine receptor (DR) agonists are similar to dopamine. They can directly stimulate postsynaptic dopamine receptors, produce the same effect as dopamine, delay the application of levodopa as much as possible, and reduce complications caused by long-term use of levodopa. Therefore, screening effective dopamine receptor agonists has become a key issue in the study and treatment of PD. In order to establish a rapid, stable and reliable method for dopamine receptor agonist screening, this study used the human dopamine receptor 2 (DRD2) gene fused with a circular permuted EGFP (cpEGFP) to construct a recombinant gene, packaged with lentiviral vector, and the vector replaced the parted inner transmembrane domain of the third intracellular loop (ICL3) of genetically-encoded GPCR-activation based (GRAB) sensors. The fluorescence of GPCR-fused cpEGFP is regulated by conformational changes mediated by the interaction of dopamine receptor agonists with GPCRs without altering GPCR activity. The HEK293T cells were infected with viral vector, screened by puromycin to select highly expressed cells. Dopamine receptor agonists (including dopamine, bromocriptine mesylate, cabergoline, pramipexole) were used as positive drugs to explore the best screening and detection conditions, establishing a stable model to evaluate the dopamine receptor agonist. The results showed that the optimal filter for the dopamine receptor agonist in this study was the cell seeding count of 7×10⁴, and the effective concentration of the positive drug was 1-100 µmol·L^-1. In addition, pretreated with 10 µmol·L^-1 dopamine receptor antagonists (including chlorprothixol hydrochloride, domperidone, and sulpiride), the positive fluorescence signal of overexpressed DRD2-cpEGFP HEK293T cells could not be detected when exposed to 10 µmol·L^-1 dopamine receptor agonists, which proved that dopamine receptor antagonists could block the activity of dopamine receptor agonists, so they cannot activate dopamine receptor allosteric, indicating that the model has good specificity and can also be used for the screening and detection of new dopamine receptor antagonists. In summary, the study constructs a stable dopamine sensor detection system, which can effectively screen potential dopamine receptor agonists. The operation procedures are simple and rapid. And it can be used for a large-scale screening providing a fundamental methodology for drug development and PD treatment targeted on DRD2.

AIMTo study the effect of proanthocyanidins on the COX-2 enzyme activity and COX-2 protein expression in LPS-induced RAW264.7 cells.

METHODSAfter being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effect of proanthocyanidins on the activity of COX-2 enzyme in RAW264.7 cells was analysed by radioimmunoassay (RIA). After being pretreated with different concentrations of proanthocyanidins for 30 min, and then 1 mg x L(-1) LPS for 9 h, the effects of proanthocyanidins on the expressions of COX-2 mRNA and protein in RAW264.7 cells were analysed by RT-PCR and Western blotting.

RESULTSThe activity of COX-2 enzyme was not inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1), P > 0.05 vs LPS group), but the activity of COX-2 enzyme was significantly inhibited by 10 micromol x L(-01) NS-398 (P < 0.01 vs LPS group). The expression of COX-2 mRNA was inhibited by proanthocyanidins (0. 8, 4 and 20 mg x L(-1)). The expression of COX-2 protein was inhibited by proanthocyanidins (4 and 20 mg x L(-1)).

CONCLUSIONProanthocyanidins had no effect on the activity of COX-2 enzyme in LPS-induced RAW264. 7 cells. Proanthocyanidins inhibited significantly the expression of COX-2 mRNA and protein in LPS-induced RAW264.7 cells.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Lipopolysaccharides ; Macrophages, Peritoneal ; cytology ; enzymology ; Mice ; Proanthocyanidins ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics

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

Objective To screen out specific microsatellite markers for use in Tupaia belangeri chinensis genetic testing. Methods Firstly to screen about 700 microsatellite loci from whole genome.Secondly to choose about 100 better loci without defect factors.Lastly 46 primers were designed by 33 tree shrew’ s microsatellite loci obtained from whole genome and other references.Agarose gel electrophoresis and polyacrylamide gel electrophoresis were used for PCR products, and better loci based on electrophoresis results were chosen.Then STR scan was used to select the microsatellite loci combination for genetic testing.Results Twenty-two microsatellite loci were selected with a significant Stutter peak on STR scanning.Comparing the alternative loci and ultimately selected loci, there were two loci available in the five alternative loci of T.glis.The coincidence rate between T.glis and T.b.chinensis was 40%.There were two loci available in the five alternative loci of T.minor, and the coincidence rate between T.minor and T.b.chinensis was 40%.There were two loci available in the three alternative loci of T.belangeri, and the coincidence rate between T.belangeri and T.b. chinensis was about 70%.Conclusions The 22 microsatellite loci screened in this study are well applied for genetic testing of Tupaia belangeri chinensis, therefore, provide a scientific basis for the genetic quality monitoring of tree shrews.

Objective To explore the feasibility and safety of early ambulation in patients with acute deep ve-nous thrombosis. Methods The literatures about early ambulation for acute venous thrombosis were collected and the quality of the literature was evaluated by two investigators independently. Results Thirteen published studies were recruited for systematic review,including 10 RCTs and 3 CCTs. The meta-analysis showed that compared with bed rest,early ambulation did not increase the incidence of secondary pulmonary embolism in patients with acute DVT[early activity:22/1408;bed rest:34/1417;RR=0.62,95%CI(0.37,1.03),and the incidence of DVT progression [early activity:48/1523;bed rest:45/1525;RR=0.85,95%CI(0.58,1.24)]. There were no statistically significant differ-ences in mortality,swelling,and pain relief in acute DVT patients. Conclusion Early ambulation did not increase the incidence of thrombus progression and secondary pulmonary embolism in patients with acute DVT compared with bed rest. Besides,early ambulation cannot relieve symptoms of swelling and pain in DVT patients. However,it can relieve the acute pain of DVT patients with moderate to severe pain.