13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

Background Recent studies have demonstrated that epicardial flow in nonculprit arteries,which has been assumed to be normal,was slowed in the setting of ST-elevation myocardial infarction (STEMI).However,the impact of primary percutaneous coronary intervention (PCI) on blood perfusion in nonculprit arteries in patients with STEMI has not been clarified.The purpose of this study was to investigate the impact of primary PCI on blood perfusion in nonculprit arteries in patients with STEMI and correlated clinical factors.Methods A total of 117 patients with anterior wall STEMI,the culprit artery being the left anterior descending artery (LAD),undergoing primary PCI (the study group) and 100 patients with normal coronary angiography (the control group) were enrolled.To observe the differences of corrected TIMI frame count (cTFC) and myocardial blush grade (MBG) before and after primary PCI in both culprit and nonculprit arteries,the left circumflex coronary artery (LCX),cTFC and MBG in the LAD and LCX were measured in the study group and control group.The study group was divided into three groups; reflow in the culprit artery group (the R group),no reflow in culprit artery group (the NR group),and no reflow in both the culprit artery and nonculprit artery group (the NRB group) according to MBG grade.The level of serum C-reactive protein (CRP),catecholamine,and fibroblast growth factor-21 (FGF21) were assayed.The clinical and angiographic characteristics were also analyzed.Results cTFC (28.1±24.3 vs.20.3±19.3,P ＜0.05) and MBG in the LCX were different in the study group compared to the control group before primary PCI.cTFC (25.2±22.3 vs.28.1±24.3,P ＜0.05) and the MBG level in the LCX were improved after successful primary PCI,but were not recovered to the normal level.Patients with no reflow in the culprit artery had a higher incidence of no-reflow in the nonculprit artery (78％ vs.19％,P ＜0.0001),and the levels of CRP ((3.29±1.31) mg/dl vs.(2.51±1.14) mg/dl vs.(2.93±1.07) mg/dl,P ＜0.05,respectively),catecholamine ((epinephrine (693.48±89.78) pg/ml vs.(398.12±93.28) pg/ml vs.(562.54±96.22) pg/ml,P ＜0.0001,respectively),and norepinephrine ((7012.43±932.47) pg/ml vs.(4012.34±814.16) pg/ml vs.(5549.03±912.65) pg/ml,P ＜0.0001,respectively)) in the NRB group were higher than those in the R group and NR group.The level of FGF21 ((0.299±0.093) ng/ml vs.(0.612±0.071)ng/ml vs.(0.428±0.074) ng/ml,P ＜0.0001 respectively) in the NRB group was lower than that in the R group and NR group.Conclusions The blood perfusion in the nonculprit artery may be impaired in patients with STEMI.Although nonculprit artery perfusion may be improved after successful primary PCI,it is still lower than that in the control group,and may be involved in inflammation and spasms.

Objective To analyze the clinical characteristics and the outcome of severe trauma of various ages in emergency department,to improve the therapeutic effect of emergency rescuing.Methods Two thousand and sixty nine patients of severe trauma were treated during a seven-year period from October 1997 to October 2004.The studied patients were divided into 5 groups:(1)adolescent group(Group A,＜13 years old,n=106);(2)juvenile group(Group B, 13-18 years old,n=128);(3)youth group(Group C,18-40 years old,n=1518);(4)middle age group(Group D, 40-60 years old,n=215);and(5)elderly group(Group E,＞60 years old,n=102).All the patients were evaluated with Injury Severity Score(ISS),and the result was≥16 in all of the studied patients.Results The incidence of severe trauma in male in all 5 groups was higher than that in female,and it was significantly higher in Group C,Group D and Group E when compared with that in the other groups(P＜0.01).Traffic accident was the leading cause of injury,and its incidence in Group A,Group B and Group D was higher than that in other groups(P＜0.01).However,injury caused by falling from high places was the second cause of injury,being significantly increased in Group A,while armed fighting and injury during work being significantly increased in Group B,Group C and Group D,slip fall injury being significantly increased in Group E(P＜0.01).Head injury was mainly found in Group A and Group E,extremities injury and/or spinal injury were increased markedly in Group B,Group C and Group D(P＜0.05),abdominal injury was significantly decreased in Group E(P＜0.001).The total mortality was 11.9%(246/2069).The mortality within 24 hours (20.6%,21/102)was significantly higher than that beyond 24 hours(7.8%,8/102)in Group E(P＜0.01).The time of staying in the emergency department differed significantly between the survived patients and those died in all 5 groups (P＜0.01).Conclusion The gender,the incidence and the characteristics of the injury causes and injury sites differed between severely injured patients of various ages.Mortality in the elderly is significantly increased following severe trauma. The idea of“golden one hour”and“platinum ten minutes”,measures of shortening the time of staying in the emergency clinic,early definite operation and damage control operation should be emphasized.

Background Bevacizumab is primarily aimed at pathologic angiogenesis for off-label uses such as the treatment of ocular neovascular disorders.However,as a new anti-fibrotic and anti-angiogenic agent following trabeculectomy,the safety and efficacy of bevacizumab by multiple-time and high-dose subconjunctival injection are still under study.Objective This study was to assess the safety and efficacy of bevacizumab after multiple-time and high-dose subconjunctival injections.Methods Regular trabeculectomy filtration surgery was performed on both eyes of 18 clean New Zealand White rabbits 0.1ml of bevacizumab(25g/L) was subconjunctivally injected intraoperatively and 3,5,7 days postoperatively in the left eyes of rabbits,and no any intervene in the right eyes were as normal controls.Bleb morphology was examined every 2 days and graded based on Moorefield's criteria and compared between the bevacizumab-treated eyes and normal saline(NS) eyes.The animals were sacrificed at 10,20 and 30 days after surgery respectively.The histopathological changes of the blebs were detected by hematoxylin and eosin stain to evaluate the cellular element around the bleb,and Masson stain was used to assess the degree of fibroblast proliferation.The degree of vascularity of bleb was identified by anti-vWf stain.Approval of this protocol was obtained and permitted from People's Hospital Institutional Animal Care and Use Committee of Peking University.The use of experimental animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Compared to the NS-treated eyes,bevacizumab-treated eyes showed the larger and more diffusely elevated blebs with the significant difference(2.48±0.22cm2 versus 1.73±0.27cm2,t=5.194,P＜0.05).The survival time of the filtration bleb in bevacizumab panel was longer in bevacizumab-treated eyes compared to control eyes,showing a significantly difference between them(21.0±1.56 days versus 12.5±1.97 days,t=3.830,P=0.005).Histological and immunohistochemical analysis confirmed that bleb and adjacent conjunctiva vascularity(A value) was significantly less in bevacizumab-treated eyes than that in control eyes at 20 days after surgery with the difference value 14320.7±4134.9(t=12.275,P＜0.05),and fibroblast deposition value was evidently diminished after bevacizumab treatment at 30 days following surgery in comparison with control eyes with the mean difference 0.27±0.03(t=15.980,P＜0.05＝.Conclusion Repeated subconjunctival injection of bevacizumab can effectively prolong the survival time of bleb in a rabbit model of trabeculectomy and limit the degree and area of vascularization in 30 days following surgery.Bevacizumab inhibit fibroblast-meditated tissue formation significantly in the later phase of vascularization after trabeculectomy.

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Membrane ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Signal Transduction ; TCF Transcription Factors ; metabolism ; Transcription Factor 7-Like 2 Protein ; Wnt Proteins ; physiology ; beta Catenin ; metabolism

The effect of the complement C1q expression on total hepatic ischemia-reperfusion (I/R) injury in rats was investigated. Sixty healthy male Sprague Dawley (SD) rats weighing 180-200 g were randomly divided into 5 groups: sham-operation group (S group, n=12); group of I/R for 1 h (I/R 1 h group, n=12); group of I/R for 3 h (I/R 3 h group, n=12); group of I/R for 6 h (I/R 6 h group, n=12); group of I/R for 24 h (I/R 24 h group, n=12). The hepatic I/R model of rats was established, and liver tissues were obtained 1 h, 3 h, 6 h and 24 h after hepatic I/R, respectively. Furthermore, the tissues were stained using hematoxylin-eosin, and the liver injuries of rats were observed using a microscope. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in liver tissue were determined. Real-time polymerase chain reaction (PCR) and Western blotting were used to detect the expression levels of C1q mRNA and protein, respectively. As compared with the S group, the histopathological changes in I/R 1 h-24 h groups were gradually aggravated with the extension of I/R time. As compared with the S group, SOD activity and MDA content in the I/R groups were reduced and increased respectively with the extension of I/R time (P<0.01). Furthermore, the C1q expression at mRNA and protein levels in the I/R groups (especially in the I/R 3 h group) was significantly higher than that in the S group (P<0.05). It is suggested that C1q expression may play a principal role in hepatic I/R injury, particularly at the early stage of perfusion.

Objective :To fabricate resveratrol nanoemulsions and probe their pharmacokinetics profiles in rats in vivo. Methods The nanoemulsions of resveratrol were tailored by using aethylis oleas as the oil phase, Cremophor RH40 as emulsifiers, absolute alcohol as co-emulsifiers, respectively. The formulation was optimized by pseudo-ternary phase diagrams and their physicochemical properties of nanoemulsions were characterized by particle size distribution, transmission electron microscope (TEM), and Fourier transform infrared spectroscopy (FTIR). The pharmacokinetics experiments were also performed in rats after gavage and the plasm concentration of resveratrol were determined by HPLC as well as the profiles of pharmacokinetic parameters were obtained by Drug and Statistics (DAS) software. Results: The formula of nanoemulsions were as follows: the ratio of resveratrol-aethylis oleas-mixture surfactant- distilled water was 1:10:24:65. The average particle size diameters of prepared resveratrol nanoemulsions were about 40 nm and the droplets of nanoemulsions were in spherical shape observed by TEM. The results of IR disclosed that the active trans-resveratrol presented in the oil droplet of the nanoemulsions. Compared with resveratrol suspensions, the bioavailability of resveratrol nanoemulsions was increased 1.45-fold, and Cmax reached to 1.93-fold. Conclusion : These results indicated that nanoemulsions may be a promising tool for the delivery of resveratrol.

Objective To evaluate the clinical features and the strategy for the diagnosis and treat- ment of bilateral testicular tumor.Methods The clinical data (including the signs and symptoms,imaging studies,tumor markers,treatment modalities and histopatbologic diagnoses) of 10 cases of bilateral testicular tumor from January 1980 to December 2004 were reviewed.Their age ranged from 19 to 58 years(mean,34 years).Of the 10 cases,8 with metachronous and 2 with synchronous testicular tumors were identified.The clinical stages at the primary and secondary tumor diagnosis were:5 cases of stageⅠ,3 of stageⅡ;and 6 cases of stageⅠ,1 of stageⅡ,and 1 of stageⅢ,respectively,in 8 metachronous tumor patients.Two syn- chronous tumor patients were both identified as stageⅠdisease.Histological examination showed the primary tumor (seminoma) in 4 cases and the secondary contralateral tumor (seminoma) in 3.Results Two syn- chronous tumor patients underwent bilateral radical orchiectomy simultaneously,and 8 underwent orchiectomy successively.Retroperitoneal lymph node dissection was performed in 3 cases.Postoperatively,hypogonadism occurred in 10 patients,and 7 of them received androgen replacement therapy.Follow-up ranged from 9 month to 23 years with a mean of 10.5 years.Two patients died of the disease;2 had metastasis (1 of them was alive with metastasis);2 had recurrences and underwent local resection.Conclusions Metachronous bilateral testicular cancers are more common than synchronous bilateral testicular cancers.Seminoma was the most common histopathologic type.Testis-sparing surgery can be performed in selected cases.

OBJECTIVETo study the clinical features, treatment and diagnosis of parapelvic cyst.

METHODSTwenty-three patients of parapelvic cyst of the kidney were reviewed retrospectively. Fourteen cases (61%) complained of lumbar pain or discomfort, and 4 patients (17%) accompany hematuria and hypertension.

RESULTSIn 15 patients receiving surgery, 2 were treated by nephrectomy, one by radical nephrectomy for misdiagnosis. Postoperative diagnosis confirmed a cyst. Eight patients were treated conservatively for cyst being small and without clinical symptoms. Nineteen cases were followed up for 0.5 - 12.0 years.

CONCLUSIONSUltrasonography and CT scan are the main diagnostic methods. Enhanced CT is extremely helpful in differential diagnosis of hydronephrosis. Surgical management is suitable for big cysts, lumbar pain, hematuria, hypertension and other complications.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases, Cystic ; diagnosis ; etiology ; surgery ; Kidney Pelvis ; Male ; Middle Aged ; Prognosis ; Tomography, X-Ray Computed

Amyloid beta-Protein Precursor ; analysis ; physiology ; Animals ; Cell Line, Tumor ; Cytoskeleton ; chemistry ; Mice ; Neurites ; physiology ; Neuroblastoma ; pathology ; Neurofilament Proteins ; analysis ; genetics ; RNA, Messenger ; analysis