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.

Objective To explore the possible mechanism and protective effect of Xuebijing injection (血必净注射液)and dexamethasone on rats with paraquat-induced chronic pulmonary injury.Methods Thirty male Wistar rats were randomly divided into six groups:normal group(n=5),model group(n=5), treatment groups(n=20).In the normal group,normal saline was used,while in the other groups,20% paraquat 80 mg/kg was injected peritoneally for poisoning.After 2 hours of intoxication,low dose Xuebijing injection(1.25 g/kg),high dose Xuebijing injection(2.50 g/kg),dexamethasone(25 mg/kg),high dose Xuebijing injection combined with dexamethasone(combined group)respectively were administered into the four different treatment groups,equal amount of normal saline was given to the normal and model groups,and the treatment continued for 4 days.At 28 days after paraquat injection,5 rats in each group were killed respectively,serum transforming growth factor-?1(TGF-?1)and hydroxyproline(HYP)level in the lung homogenate were measured,and pulmonary coefficient and histological changes were observed.Results In the treatment groups,the levels of serum TGF-?1 and lung tissue HYP,pulmonary coefficient were leas than those of model group,and among the treatment groups,combined group had the best results(all P

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.

OBJECTIVETo systematically review the effectiveness of minimally invasive total hip arthroplasty (MIS-THA) versus traditional total hip arthroplasty (THA) in patients with hip diseases.

METHODSThrough a method of combining Free words and keywords,we searched databases including PubMed,The Cochrane Library, EMbase,Web of Science, CBM , CNKI and Wanfang Data for randomized controlled trials (RCTs) on the comparison between MIS-THA and THA for hip disease from inception to June, 2014. Two reviewers independently screened literatures according to the inclusion and exclusion criteria, extracted data and assessed the quality of the included studies according to the "bias risk assessment" tool recommended by Cochrane Handbook 5.0 for Systematic Reviews. Then, meta-analysis was performed using RevMan 5.3 software.

RESULTSThirteen RCTs involving 1 213 cases of surgeries and total 1 284 hips (MIS-THA: n = 631; THA: n = 653) were identified. The results of meta-analysis showed that statistically significant differences were found in Harris hip score on the 3rd month after operation [MD = 8.37, 95% CI (6.02,10.72)], Hematocrit [MD = 0.02, 95% CI (0.01, 0.03)] and Hemoglobin [MD = 0.50, 95% CI (0.16, 0.85)] at the 48th hour after operation, changed value of femoral offset [MD = 0.30, 95% CI (0.04, 0.56)] between two groups. In the change value of femoral offset, THA was better than MIS-THA; There were no statistically significant differences between two groups in Harris hip score at 1st year after operation [MD = 3.26, 95% CI (-3.25, 9.76)], WOMAC score [MD = -0.53, 95% CI (-3.67, 2.60)] and Oxford score [MD = 1.34, 95% CI (-3.46, 6.13)] at the 6th week after operation, Hematocrit at the 8th hour after operation [MD = -0.01, 95% CI (-0.02, 0.00)], the incidence of hip varus [RR = 0.82, 95% CI (0.45,1.52)] and dislocation [RR = 1.40, 95% CI (0.48, 4.12)].

CONCLUSIONTHA brings less trauma, less hemorrhage and better early clinical outcome compared with MIS-THA, but the difference of the complication rates between the two groups is similar.

Arthroplasty, Replacement, Hip ; methods ; Humans ; Minimally Invasive Surgical Procedures ; methods

ObjectiveTo investigate the association between the common variant rs1512268 single nucleotide polymorphism(SNP) of NKX3.1 gene and the risk of prostate cancer,and to explore its interaction with related risk factors.MethodsTotally 122 patients with prostate cancer and 105 age matched male people (prostatic specific antigen ＜ 4 μg/L,without family history of prostate cancer) as control group were enrolled.Polymerase chain reaction - high resolution melting curve(PCR - HRM) combined with gene sequencing methods were used to determine the distribution of allele and genotype frequencies of the rs1512268 SNP.ResultsThe distributions of GG,AG,AA genotypes were 42 cases(33.4％),66 cases(54.1％),14 cases(11.5％) in patients with prostate cancer,and 45 cases(42.9％),51 cases(48.6％),9 cases(8.6％) in healthy control,respectively.There were no significant differences in the distribution of genotype(x2 =1.70,0.69,0.52) and allele frequency (x2 =1.575) between the two groups(P＞ 0.05).The different genotypes of rs1512268 of NKX3.1 gene were not associated with age,Gleason score,PSA levels and clinical stage of prostate cancer (P＞0.05). Conclusions rs1512268 SNP of NKX3.1 gene is not obviously associated with prostate cancer and may be not the genetic risk factor in Chinese.

Objective To investigate the reversal effect of MDR1 and MDR3 gene silencing on resistance of A2780/taxol cells to paclitaxel.Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 genes was transfected into A2780/taxol cells.The early stage cell apoptosis and the effect of intracellular rhodamine 123(Rh123)accumulation were detected by flow cytometry(FCM).The late stage cell apoptosis rate was detected by terminal deoxynucleotidyl transferase(TdT)-mediated deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL).The 50% inhibition concentration(IC_(50))of paclitaxel on A2780/taxol cells was determined by methyl thiazolyl tetrazolium(MTT)assay.MDR1 and MDR3 mRNA were assessed by RT-PCR,and caspase-3 protein was detected by western blot.Results After treatment with MDR1 and MDR3 shRNA plasmid vector,early apoptosis rate of A2780/taxol cells was (20.21?0.56)% and(10.87?1.24)%,respectively.MDR1 and MDR3 shRNA could increase cellular Rh123 accumulation(116.6?8.1 and 98.4?3.8,respectively).The late stage apoptosis rates detected by TUNEL displayed the same tendency as FCM results did.The IC_(50)for paclitaxel of A2780/taxol cells was decreased significantly.The mRNA levels of MDR1 and MDR3 in A2780/taxol cells were decreased by (73.3?0.8)% and(51.6?0.4)% of control,and the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner.The expression of caspase-3 protein of MDR1 and MDR3 shRNA vector transfected group in A2780/taxol cells was significantly increased [(80.8?2.6)% and(72.0?4.7)%, respectively ].Conclusion MDR1 and MDR3 gene silencing could recover sensitivity of A2780/taxol cells to paclitaxel and induce cell apoptosis,thus reversing cell resistance to paclitaxel.

Objective To determine levels of serum Leptin,insulin like growth factor-1(IGF-1),interleukin-6(IL-6) and tumor necrosis factor-alpha(TNF-?) in newborn infants with hypoxic-ischemic encephalopathy(HIE).Methods The asphyxiated and normal term neonates were included.The HIE group contained 45 cases and control group 20 cases.Serum Leptin,IGF-1,IL-6 and TNF-? levels were measured by a sensitive enzyme-linked immunosorbent assay.Results In asphyxiated term neonates,serum Leptin,IGF-1,IL-6 and TNF-? levels were significantly higher or lower than those in control group(all P

OBJECTIVETo evaluate the characteristics of the preputial development in Chinese boys and indications as well as occasion of circumcision.

METHODSThe shape and retractability of prepuce were evaluated in 1,015 Chinese boys from 0 to 18 years old without heteroplasia of prepuce and penis.

RESULTSRatio of children with phimosis decreases progressively with increasing age, from 64.09% in the first group (0 approximately 3 years old) to 7.66% in the fourth group (11 approximately 18 years old).

CONCLUSIONPhimosis and incomplete separation of the prepuce is normal in the neonate and infant. Prepuce will separate from the glans progressively till adolescence, so it's unnecessary to perform circumcision for them when without any complications.

Adolescent ; Child ; Child, Preschool ; China ; Circumcision, Male ; Contraindications ; Humans ; Infant ; Infant, Newborn ; Male ; Penis ; anatomy & histology ; Phimosis ; pathology

Objective To assess the changes on the whole night polysomnography (PSG) in patients with chronic fatigue syndrome(CFS). Methods The whole night PSGs were recorded from 24 patients with CFS and 33 normal subjects. Results Compared with normal subjects, patients with CFS showed significantly reduced total sleep duration ([488.7?21.7]min vs [515.9?31.7]min, P

Objective To study the purification and renaturation of recombinant human IL-2/GM-CSF fusion protein. Methods After the inclusion bodies were isolated, the optimal conditions of denaturation and renaturation were studied. The renatured recombinant human IL-2/GM-CSF was purified by DEAE-Sepharose FF ion-exchange chromatography. Results The purified product manifested biological activity with purity of up to 95%. Conclusions The procedures employed in this study is simple and reliable.