Objective:The aim is to establish L-glutamate specific aminotransferase-L-glutamate dehydrogenase coupling 96-well high throughput screening method,which is applied to molecular evolution of aminotransferase WecE from E.coli.Methods:An optical assay for aminotransferase catalytic activity based on aminotransferase-glutamate dehydrogenase coupling system is established by optimization of coupling enzyme loading,signal molecule NADH concentration and coupling time.Mutants library of WecE is obtained by sitedirected saturation mutagenesis.Positive mutants can be screened out through 96-well preliminary screening and flask second screening.Results:The target transamination reaction is coupled with L-glutamate dehydrogenase indicative reaction system which consists of 0.5 U/ml enzyme loading and 0.4 mmol/L NADH.A positive mutant Y321F whose catalytic activity increases 3.4 fold compared to that of wild type is screened out in Tyr 321 saturation mutagenesis library of WecE.Conclusion:An accurate high throughput screening method with weak background interference is established.It offers feasible solution for molecular evolution of L-glutamate specific aminotransferase.

Objective To evaluate endovascular therapy and open surgery for subclavian artery occlusion disease. Methods In this study, 69 patients received endovascular therapy (44 patients)or open surgery(25 patients)from January 2002 to July 2007.Balloon dilatation was carried out in 3 cases and 43 stents was placed in 41 cases. Results All procedures were successful. In endovascular therapy group, the ratio of healthy/dieased side of mean blood pressure was improved from 0.66±0.14 to 0.96±0.13(t=9.532,P＜0.001＝;in surgery group, the ratio improved from 0.63±0.16 to 0.95±0.18(t=8.236,P＜0.001＝.Sixty-one discharged patients were followed up for 2～49 months(mean 16.7 months),in endovascular group, restenosis occurred in 1 patient 1 year after the therapy, in surgery group, all prothesis remained patent and there was no complication related to prothesis. Conclusions Both endovascular therapy and surgery were the effective methods for subclavian artery occlusion, and endovascular therapy is preferred for less invasiveness.

OBJECTIVETo investigate the potential developmental toxicity of Radix Ophiopogonis decoction in SD rats.

METHODTimed-pregnant SD rats were given Radix Ophiopogonis decoction (26.9 g x kg(-1)) or vehicle (distilled water) by gavage on gestation days 6-17. Maternal clinical sign, abortions, premature deliveries, and body weight were monitored throughout gestation. At termination (gestation days 20) pregnant females were evaluated for clinical status and gestational outcome; live fetuses were examined for gender, external, visceral and skeletal malformation and variations.

RESULTNo deaths, premature deliveries or dose-related clinical signs were attributed to Radix Ophiopogonis decoction. Maternal body weight and body weight gain were not affected. There were no effects on fetus weight and viability, incidences of fetal malformation and variation.

CONCLUSIONThese results demonstrated that Radix Ophiopogonis decoction had no detectable adverse effects in either the treated F0 female rats or the fetuses.

Animals ; Body Weight ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Embryonic Development ; drug effects ; Female ; Fetal Development ; drug effects ; Fetal Weight ; drug effects ; Humans ; Male ; Models, Animal ; Ophiopogon ; chemistry ; Pregnancy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

ObjectiveTo investigate the clinical value of color Doppler ultrasound examination in the diagonosis of acute and chronic artery occlusion of the extremities.MethodsA review was made on 129 extremetiy artery occlusion patients at Anzhen Hospital during 2006 -2010. 85 cases were male, and 44 cases were female. Age was from 17 to 94 years (average: 62 ±9 years). We analyzed two-dimensional and color Doppler flow imagings of 39 acute occlusion arteries and 97 chronic occlusion arteries. We compared factors including the echoes of artery lumens, the vessel wall structures, hemodynamic parameters of inlet and outlet at the occlusion, and collaterals between groups.ResultsThe factors of depths of vessel wall,internal diameters of ccclusion arteries, proximal resistant index and collaterals were significantly different between groups ( P ＜ 0. 05 ). The internal diameters of acute occlusion arteries were wider than chronic occlusion arteries. The depths of vessel wall, proximal resistant index and collaterals were thinner, smaller,and less than chronic occlusion arteries. The total accurate rate of differential diagnosis for acute and chronic artery occlusion by color Doppler ultrasound was 95.6％.ConclusionsColor Doppler ultrasound is an effective method for the differential diagnosis of acute and chronic artery occlusion of the extremities.

Objective To improve the quality of the management of the elderly patients with cancer in Sichuan province,and to understand the attitudes and perspectives of elderly surgeons and oncologists for the management and treatment of elderly patients with cancer.Methods A face-toface questionnaire interview was conducted with oncologists (n 64) and geriatricians (n =64).128 physicians were involved in this study.Results The cancer management and therapeutics were deemed appropriate at present by 9.38％ (6/64)of the geriatricians and 25.00％ (16/64)of the oncologists.The 39.06％ (25/64) of geriatricians used to notice geriatric syndromes,while 81.25％ (52/64)of oncologists never concerned about the geriatric syndrome(P=0.011).As for the causes of the therapy-associated toxicity,oncologist versus geriatrician payed an attention to malnutrition (100.00％ vs.100.00％ in both groups),to mobility disorders(65.63％ vs.65.63％,84/128 in both groups),to cognitive impairment/mood disorder (89.06％ in geriatrician group vs.75.00％ in oncologist group,P=0.038).For the factors affecting treatment decisions,a physical ability attention (oncologists vs geriatricians:70.31％ vs.92.19％;P =0.002),and comorbidity (oncologists vs geriatricians:62.50％ vs.79.69％,P =0.032) had statistically significant difference.In addition,lack of geriatrics knowledge was also reported by more oncologists.However,one hundred percent of participants wanted very much to cooperate with each other in their clinical work.When responding to the clinical scenario,the 10.94％ (7/64)of geriatricians and 32.81％ (21/64)of oncologists chose modified treatment for 65-74 years old patient with cancer(P =0.003).When the age of the patients was 75-84 years old,only 12.50 ％ (8/64) of geriatricians prefer end-of-life care,while 31.25 ％ (20/64) of oncologists chose it (P =0.010).Conclusions Selection of treatment decisions in the elderly patients with cancer affect by ageing.Both oncologists and geriatricians are concerned with the elderly patients with cancer,ageing syndrome,total sickness and functional status.And these doctors support an establishment of a multi disciplinary team cooperation for the elderly patients with cancer.Therefore,the establishment of mutual cooperation between the two professionals is necessary and feasible.

Objective To evaluate the perioperative and long-term effects of endovascular aneurysm repair(EVAR) of infrarenal abdominal aortic aneurysm (AAA).Methods Clinical data of 131 AAA cases undergoing EVAR were retrospectively evaluated for the safety and long-term efficacy.Results The operative time was (137 ±29) min,blood loss was (142 ±20) ml,blood transfusion was (46 ± 26) ml,ICU staying time was (17 ± 4) h.Major perioperative complications were severe heart failure in 8 cases,myocardial infarction in 2 cases,pulmonary complications in 5 cases,internal leakage in 4 cases.During the period of up to 60 months there were15 cases of endoleak including 8 cases of type Ⅰ,5 cases of type Ⅱ,1 each case of type Ⅲ and Ⅳ and 2 deaths.By Kaplan-Meier survival analysis there were complications developing after 60 months and up to 40％ of them needing reintervention.Conclusions Endovascular repair is the safe treatment for AAA,but discharged patients need close long-term follow-up.Complications that ensued need intensive management.

Objective To investigate the clinical curative effect of the surgical method of transferring the radialis proper digital nerve of damaged index finger and its dorsal branch to repair the thumb nerve evulsion.Methods 13 patients with thumb nerve evulsion were treated.There were 8 males and 5 females with an average age of 28 years (ranged 18-52 years old).The injuries were caused by machine twist (8 cases),gear(4 cases),electric saw (1 case).And thumb rotational avulsion amputation (10 cases),thumb incompleteness amputation(3 cases).The time from injury to admission was 1-3.5 h (mean 2.2 h).The average time from injury to admission was (1.1 ± 1.5) h.The amputate level of skin was at the juxtra-articular of metacarpophalangeal joint.The amputate level of bone was at the base of proximal phalanx (6 patients) and metacarpophalangeal joint (4 patients),interphalangeal joint(3 patients).Using transferring the radialis proper digital nerve of index finger without reimplantation and its dorsal branch to.repair the both side injuries of thumb nerve evulsion.According to routine method to repair digital bone,veins,arteries and tendons.Results All 13 chases were followed up for 6 months to 2 years and 7 months postoperatively,with an average of 22 months.The wounds and incisions at donor sites were healed by first intention.All 13 cases of thumb reimplantation were successful.Two-point discrimination of ulnaris finger pulp was 2 to 6mm,average 4.2mm,and the radialis was 5 to 9mm,average 7.8mm.Sensory function was rated as S4(the ulnaris in 12 cases and the radialis in 2 cases) and S3 + (the radialis in 11 cases and the ulnaris in 1 cases).Conclusion Transferring the radialis proper digital nerve of damaged index finger and its dorsal branch to the digital nerve on the neighboring thumb is a simple and effective method to restore sensory function of the pulp.

Glutamate dehydrogenase (GDH)a key enzyme in the nitrogen metabolism pathway catalyzes the con-version between α-ketoglutarate and glutamate reversibly using NAD(P)H as a cofactor. Based on genomic stud-ies,it was concluded that SHJG_7666 was a potential GDH in Streptomyces hygroscopicus 5008(S5008),and its expression level in vivo was positively correlated with the biosynthesis of an important aminocyclol compound vali-damycin. Phylogenetic tree analysis showed that the S5008 SHJG_7666 GDH belonged to the Glu/Leu/Phe/Val dehydrogenase family,with conserved glutamate-α-ketoglutarate binding domain and the classical GXGXXG dinu-cleotide binding motif. Further homologous modeling and structural comparison revealed that SHJG_7666 con-tained conserved Lys60,Lys78and Asp120catalytic functional sites and ligand binding residues Ser36,Gly38,Gln119 and Asp166,Asn300,Ala330. Moreover,recombinant expression of SHJG_7666 in E. coli and in vitro enzyme activity demonstrated that glutamate dehydrogenase can convert ammonium salt to glutamate with pH and temperature being optimal at 7. 5 and 37 °C respectively. Enzyme activity under optimum reaction condition has Kmvalue of (25. 3 ±9. 1)μmol/L and kcatof (3 ±0. 8)×10 -5s-1for the substrate α-ketoglutarate. Results of this study further improved the catalytic activity of SHJG_7666,thus laying the foundation for the ultimate increase of vali-damycin production.

Objective: To observe the effect of electroacupuncture on the volume of cerebral infarction, apoptosis of cerebral cells and the expression of protein kinase A (PKA) in the cerebral cortex of rats after ischemia and reperfusion so as to explore how electroacupuncture stimulates brain protection. Methods: One hundred and twenty healthy, adult, male Sprague-Dawley rats were randomly divided into a sham operation group, a model group, an electroacupuncture group and an electroacupuncture with pre-stimulation group. All except the rats in the sham operation group received occulusion of the left middle cerebral artery using the intraluminal thread method for 2h and then reperfusion. Before the operation, the rats in the electroacupuncture with pre-stimulation group were given 30 minutes of electroacupuncture at the baihui, dazhui and right neiguan points every day for 5 days. After the operation both the electroacupuncture group and the pre-stimulated group were given that same electroacupuncture regimen. The other two groups received no special treatment. Garcia scoring was used to evaluate the neurological deficits of all of the rats 5 and 10 days after the intervention. Meanwhile, the ischemic volume, apoptosis of cortical cells and PKA-positive cells were determined using flow cytometry and immunohistochemistry after triphenyltetrazolium chloride (TTC) staining. Results: The neurological function of the injured rats was severely impaired, while no neurological deficit was found in the sham operation group. The average Garcia score, cerebral infarction volume, cerebral apoptosis rate and PKA-positive cell expression rate of the electroacupuncture and electroacupuncture with pre-stimulation groups were all significantly better than those of the model group at the same time points. The averages of the electroacupuncture with pre-stimulation group were all significantly superior to those of the electroacupuncture group at the same time points. Conclusions Pre-stimulation using electroacupuncture can promote the recovery of injured nerves after cerebral ischemia and reperfusion, at least in rats. Electroacupuncture′s protective mechanism may be related to its reducing the infarcted volume, inhibiting apoptosis of brain cells and promoting PKA expression.

Objective:To investigate the necessity of adaptive re-planning during radiotherapy for nasopharyngeal carcinoma (NPC) and its impact on dose improvement.Methods:Clinical data of 89 NPC patients admitted to Sun Yat-sen University Cancer Center from July 2014 to December 2017 were retrospectively analyzed. All patients received 25+7 rounds of adaptive re-planning during radiotherapy. Plan-A was defined as the initial CT scan-based 25-fraction radiotherapy plan, while plan-B was defined as the re-planned 7-fraction radiotherapy plan based on a subsequent CT scan. The changes in the target and parotid gland volumes were compared between plan-A and plan-B. Plan-I was a one-time simulation of plan-A extended to 32 fraction radiotherapy plan, and plan-II was generated through registration and fusion of the plan-A and plan-B for adaptive re-planning. The differences in dose metrics, homogeneity index (HI), conformity index (CI), and dose to organs at risk (OAR) were compared between plan-I and plan-II. Statistical analysis was performed by using paired t-test. Results:Compared with plan-A, the gross tumor volume of massive bleeding lesions (GTV nx) and parotid gland volume of plan-B were decreased by 13.14% and 11.12%, respectively (both P<0.001). While planning clinical target volume of metastatic lymph nodes (PCTV nd) of plan-B was increased by 7.75%( P<0.001). There were significant changes in the lymph nodes of plan-A and plan-B. The D mean, D 5%, D 95% of massive bleeding lesions planning target volume (PTV nx) and D 5% of high risk planning target volume (PTV1) in plan-II were all significantly higher than those in plan-I (all P<0.05). The CI of PTV nx and PTV1 in plan-II was closer to 1 than that in plan-I. In all assessed OAR, the D mean, D 50%, and D max of plan-II were significantly lower than those of plan-I (all P<0.05). Conclusions:During radiotherapy, NPC patients may experience varying degrees of primary tumor shrinkage, parotid gland atrophy, and lymph node changes. It is necessary to deliver re-planning and significantly improve the dose of target areas and OAR.