The phenylacetone monooxygenase, isolated from Thermobifida fusca, mainly catalyzes Baeyer-Villiger oxidation reaction towards aromatic compounds. Met446 plays a vital role in catalytic promiscuity, based on the structure and function of phenylacetone monooxygenase. Mutation in Met446 locus can offer enzyme new catalytic feature to activate C-H bond, oxidizing indole to finally generate indigo and indirubin, but the yield was only 1.89 mg/L. In order to further improve the biosynthesis efficiency of the whole-cell catalyst, metabolic engineering was applied to change glucose metabolism pathway of Escherichia coli. Blocking glucose isomerase gene pgi led to pentose phosphate pathway instead of the glycolytic pathway to become the major metabolic pathways of glucose, which provided more cofactor NADPH needed in enzymatic oxidation of indole. Engineering the host E. coli led to synthesis of indigo and indirubin efficiency further increased to 25 mg/L. Combination of protein and metabolic engineering to design efficient whole-cell catalysts not only improves the synthesis of indigo and indirubin, but also provides a novel strategy for whole-cell catalyst development.
Escherichia coli ; genetics ; metabolism ; Glucose ; metabolism ; Indigo Carmine ; metabolism ; Indoles ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; Metabolic Networks and Pathways ; Protein Engineering

BACKGROUND: Many methods involved in measuring energy expenditure of physical action, however, study regarding measurements of upper limber movement energy expenditure are few.OBJECTIVIE: To measure the energy expenditure of limber movement using indirect calorimetry, and to analyze the characteristics of upper limber movement energy expenditure and influencing factors such as age and gender.DESIGN, TIME AND SETTING: The comparison observation was performed at the Jiangsu Institute of Sports Science from January to March 2009.PARTICIPANTS: Totally 108 health adults, including 47 males and 61 females, participated in the study, 65 of them aged 20-39 years, and the remained 43 aged 40-59 years.METHODS: Energy expenditure of 4 upper limb movements were performed, including sitting posture arm swing (60 times/min),elbow flexion (40 times/min), shoulder flexion-extension (60 times/min), and lateral arm raise (30 times/min). At each interval,have a 3-minute rest.MAIN OUTCOME MEASURES: Oxygen expenditure of 4 kinds upper limb movements were measured by MetaMax 3B tester. RESULTS: Rest oxygen expenditure of males were greater than that of females (P < 0.05), greater in 20-39 years old than 40-59 years old (P < 0.05). The greatest oxygen consumption occurred in males with arm swing [(550.9±90.6) mL/min], which was (425.8±75.7) mL/min in females. The lowest oxygen expenditure was elbow bend [male: (440.4±82.7) mL/min, female: (367.0±60.1) mL/min]. Net oxygen expenditure of all 4 kinds of upper limb movements was lower than 250mL/min (1 metabolic equivalent).CONCLUSION: ① The energy expenditure of usual upper limber movements below 1 metabolic equivalent. ② Age and body mass has more effect on resting energy expenditure than the gender. ③ The efficiency of young adults upper limb movements is higher than old adults.

Objective To establish an accurate method for determining the content of three components in Fufang Buwu Syrup. Methods TLC scanner was selected to detect three components with silica gel G thin layer plate. The sample was separated by using cyclohexane-ethyl acetate-methylenechloride-formic acid (3∶1∶1∶0.2),λS=300 nm. Results The linearity between peak area and ferulic acid was achieved in the range of 0.36-0.84μg, psoralen was achieved in the range of 0.12-0.28 μg, emodin was achieved in the range of 0.01-0.05 μg. The average recovery was 100.7%, 100.8%, 101.0%, and RSD was 1.26%, 1.44%, 1.86%, respectively. Conclusion The method is simple and accurate, which can be used for quality control of Fufang Buwu Syrup.

Objective To explore the extent of agreement between measurements of temporal and spatial gait parameters made with portable gait analysis equipment and in the laboratory.Methods Fifteen healthy young people submitted to laboratory gait analysis using 3D motion analysis apparatus and then on the same day to analysis using the Gait Watch portable apparatus.Cadence,stride length,walking speed and step length were recorded.Intraclass correlation coefficients (ICCs) and Bland-Altman plots were used to evaluate the agreement between the two gait analyses.Results Test-retest comparisons with the Gait Watch apparatus generated ICCs for the temporal and spatial parameters ranging between 0.80 and 0.98,indicating good test-retest reliability.Bland-Altman plots comparing the two measurement systems also showed good agreement.According to paired simple t tests,the stride length,walking speed,and step length assessments with the two systems showed significant differences.All exceeded the minimum detection threshold (stride length =0.05 m,walking speed =0.12 m/s,left step length =0.03 m,right step length =0.04 m).Conclusions Measurements of cadence,stride length,walking speed and step length with the two systems yield acceptable agreement,and either can be used in clinical walking assessment.

The Hospital Survey on Patient Safety Culture (HSOPSC) questionnaire was used for appraisal in a newly-built general hospital for all the nursing staff, and they were provided with a six-month safety culture training according to appraisal results.The training has elevated the positive response rate of the nursing staff towards all dimensions of patient safety culture, proving that scientific and reasonable safety culture training is conducive to nurses' perception of patient safety culture.

Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) has become the standard first-line treatment for advanced non-small cell lung cancer (NSCLC) patients with EGFR-sensitive mutations. However, acquired resistance cannot be avoided during the targeted treatment of EGFR-TKI, which may lead to the progression of NSCLC. Among them, approximately 50% of patients present with primary lung lesions and/or initial metastastic lesion progression. As a major local treatment, radiation therapy plays an irreplaceable role in the management of patients with advanced cancer. Stereotactic body radiation therapy (SBRT), which is implemented with the Cyberknife, is the most advanced radiation therapy technology to better meet clinical needs. In recent years, based on the abovementioned patterns of tumor recurrence, the treatment mode of pulmonary-lesion SBRT combined with EGFR-TKI can improve the tumor-free survival in advanced NSCLC patients, which has attracted widespread attention. In this article, the clinical research progresses on this combined therapeutic strategy were reviewed.

Objective To investigate the microsurgical treatment of subdural extramedullary tumors in ventral spinal.Methods Retrospectively analyzed 16 cases of subdural extramedullary tumors in ventral spinal which resected by microsurgical technique from January 2000 to December 2013.By summarized the tumor character,location,extent of resection and rehef of symptoms.Results All of 16 cases were successfully resected,in which 14 cases completely resected,2 cases mostly resected.A higher response rate of local symptoms and neurological symptoms obtained postoperatively,2 patients got worse sensory disorders,movement disorder compared with preoperative,cerebrospinal fluid leakage occured in 1 patient.Conclusion Resection of the subdural extramedullary tumors in ventral spinal with microsurgical technique has the advantage of more clearly anatomical level,minimal tissue injury of spinal cord and surrounding tissue,higher tumor removal rate.

Objective To assess the clinical efficacy of minimally invasive internal fixation combined with percutaneous kyphoplasty (PKP) in treatment of thoracolumbar burst fractures in the elderly.Methods Twenty-one cases of neurologically intact thoracolumbar burst fractures treated by PKP between January 2007 and December 2008 were included in this study.There were 8 males and 13 females at age of 65-78 years (means,70.6 years).Mean period from injury to operation was (3.7 ± 1.1) days (range,3-7 days).The injured segments included Ti1 in two cases,T12 in six,L1 in eight and L2 in five.Kphosis Cobb' s angle,correction degree of kyphosis angle,correction loss of kyphosis,perioperative indicators,visual analogue scale (VAS) and Oswestry disability index (ODI) were evaluated after operation.Results All the cases were followed up for a period of 26-56 months (mean 34.2 months).Operation averaged 98.7 minutes (range,80-120 minutes) and showed mean blood loss of 32.8 ml (range,30-85 ml).Ambulation started at mean 18.2 hours after operation (range,8-19 hours).VAS averaged (0.9 ± 0.6) points at postoperative one week.Postoperative X-ray films revealed mean 10.4° correction of kyphotic Cobb angle,followed by mean 1.8 °of loss in the follow-up longer than 24 months.According to hyperextension and hyperflexion radiographies,there was no abnormal activity of fixed segments,implant loosening and breakage or adjacent vertebral fractures.Conclusion Minimally invasive internal fixation combined with PKP can relieve pain and restore function in a short time and is thought to be a preferable treatment option for thoracolumbar burst fractures in the elderly.

Objective To establish the cell line from specimens of resectable human gastrointestinal stromal tumors (GIST) and to verify the characteristics of cell biology in vitro. Methods The tissues from biopsies of human GIST were cultured in RPMI 1640 media supplemented with 10% fetal bovine serum. After growing to 90% confluence,the cells were detached for subculture and their characteristics,including morphology,growth kinetics,karyotype analysis,immunohistochemical analysis and tumorigenicity in nude mice were determined. Results GIST named GIST-H1 was successfully established. The cell line was passaged for more than 60 times 1 year. The characteristics demonstrated: The population doubling time calculated in the log phase of growth was 47.5 h. The cloning efficiency in the soft agar averaged 24.8%.Electronmicroscopically,there were rich ribosomes and mitochondrion in the cytoplasm. Immunohistochemical analysis showed CD117(+),SMA(+),dog-1(+),CD34(-),and S-100(-).Karyotype analysis illustrated aneuploidy with the modal chromosomal number 60-98.The GIST cells transplanted in nude mice had high tumorigenicity. Conclusion The immortalized GIST cells are devoloped in vitro and have specific characteristics of GIST.

OBJECTIVETo investigate the short-term clinical outcome of unicompartmental knee arthroplasty for the treatment of spontaneous osteonecrosis of the knee.

METHODSFrom September 2013 to April 2014,5 patients with spontaneous osteonecrosis of the knee underwent unicompartmental knee arthroplasty, included 3 males and 2 females, aged from 65 to 80 years old with an average of 74 years. The courses of disease was from 1 to 6 years with the mean of 3 years. According to the radiographic staging criteria of Koshino, 1 case was stage II, 2 cases were stage III, 2 cases were stage IV. Clinical effects were assessed by VAS score, HSS score, and knee range of motion, tibiofemoral angle before and after operation.

RESULTSAll the patients were followed up from 6 to 7 months with an average of 6.4 months. All incisions obtained primary healing, and there were no complications such as infection, thrombosis, fracture of lower limbs. All 5 patients' pain relieved and their knee function improved significantly after operation, but knee range of motion had no obviously improved. Postoperative HSS scores, VAS scores, tibiofemoral angle were significantly improved than that of preoperative.

CONCLUSIONThe short-term effect of unicompartmental knee arthroplasty in treating spontaneous osteonecrosis of the knee is satisfactory.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Joint Diseases ; physiopathology ; surgery ; Knee Joint ; Male ; Osteonecrosis ; physiopathology ; surgery ; Range of Motion, Articular