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

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 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

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.

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

OBJECTIVETo investigate the relationship of protein kinase C-alpha (PKCalpha) expression/activation with tumor differentiation and resistance to chemotherapy drugs in superficial bladder carcinoma.

METHODSExpression of PKCalpha was measured by Western-blot analysis in 76 samples including tumor and normal tissues, respectively. A human RT4 bladder cancer cell line stably expressing green fluorescent protein (GFP)-PKCalpha (RT4/PKCalpha) was established. The sensitivity of the RT4/PKCalpha and parental cells to adriamycin (ADM) was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The change of sensitivity of the RT4/PKCalpha to ADM were observed under the conditions of PKC activation and inhibition, respectively.

RESULTSTotal level of PKCalpha expression and the ratio of the amount of PKCalpha expression or PKC activity in membrane to that in cytosol (M/C) were all more higher in cancerous tissues than in normal tissues (P < 0.01); With the increase of tumor grade, the relative level of PKCalpha expression significantly increased in membrane (P < 0.01) and decreased in cytosol (P < 0.01), M/C of PKCalpha was significantly elevated (P < 0.01), and total relative level of PKCalpha expression significantly increased (P < 0.01). Thirty-eight cases recurred during the follow-up period in total seventy cases. Multivariate analysis showed that high M/C of PKCalpha was independent prognostic factor for tumor recurrence after standard ADM treatment in the 2-year follow-up (RR = 3.98, 95% CI 1.22-5.68, P = 0.03). Transfection of PKCalpha increased resistance of RT4 cells to ADM [resistance index (RI): 6.97, t = 3.24, P < 0.01]. PKCalpha activation further greatly promoted the resistance (RI: 148.11, t = 5.18, P < 0.001) while inhibition of PKCalpha did conversely (RI: 1.6, t = 1.29, P > 0.05).

CONCLUSIONThe abnormal activation and expression level of PKCalpha closely correlate with both tumor grade and intrinsic resistance to ADM in patients with superficial bladder carcinoma.

Aged ; Antibiotics, Antineoplastic ; pharmacology ; Carcinoma, Transitional Cell ; enzymology ; pathology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; physiology ; Enzyme Activation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Protein Kinase C-alpha ; metabolism ; Transfection ; Tumor Cells, Cultured ; Urinary Bladder Neoplasms ; enzymology ; pathology

OBJECTIVETo optimize extraction technology of the seed of Ziziphus jujuba var. spinosa with the targets of the total saponin, total jujuboside A and B and total flavonoids.

METHODIn the method of one-way and orthogonal tests, ethanol concentration, amount of ethanol, extraction time and extraction times were the factors in orthogonal test, and each factor with three levels.

RESULTEthanol concentration and extraction times had significant effect on all the targets, other factors should be selected in accordance with production practice.

CONCLUSIONThe best extraction technology is to extract for three times with 8 fold ethanol solution (60%), and 1.5 h each time.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol ; Flavones ; analysis ; Plants, Medicinal ; chemistry ; Saponins ; analysis ; Seeds ; chemistry ; Technology, Pharmaceutical ; methods ; Ziziphus ; chemistry