OBJECTIVE:To establish a HPLC method for the determination of 5-fluorouracil in human plasma METHODS:The plasma samples were extracted with ethyl acetate-isopropanol(85∶15,V/V)following precipitation of plasma proteins with ammonium sulfate The solvent was evaporated to dryness under a stream of nitrogen The dry extract was dissolved in mobile phase and then injected into the Diamonsil C18 column(150mm?4 6mm,5?m)and 0 01mol/L KH2 PO4(pH5 5) was taken as mobile phase The flow rate was 1 5ml/min,UV detection wavelength was 267nm The internal standard was 5-bromouracil RESULTS:The minimal detectable drug concentration in plasma was 0 025?g/ml The calibration curve was linear over a range from 0 3?g/ml to 10?g/ml The intra-day RSD≤5 0%(n=5) and the inter-day RSD≤11 5%(n=5) CONCLUSION:The method can be used for studying the pharmacokinetics and bioavailability of 5-fluorouracil

Objective To investigate the clinical effect of lung equivalent uniform dose (LEUD)-based predictive model for radiation pneumonitis (RP) induced by volumetric modulated arc therapy (VMAT) and to determine the optimal a value.Methods A total of 65 patients with primary lung cancer who received VMAT from July 2015 to February 2016 were divided into RP group and non-RP group according to the presence or absence of RP after radiotherapy.Their dose-volume histogram (DVH) data and other data were obtained and analyzed by the self-compiled numerical analysis program.The LEUD values in the two groups were calculated at a=[-50, 50], and then the a value was identified when the relative difference of LEUD between the two groups was maximal.The paired t test was used for analyzing the differences in V5, V20, V30, minimum lethal dose (MLD), and LEUD (aoptimal) between the two groups.A Pearson correlation analysis was used to determine the correlation of Vdose and LEUD (aoptimal) with RP.The logistic regression method was used to establish the predictive model of RP.Results The maximum relative difference in LEUD between RP group and non-RP group was obtained at a=0.3(627.94 cGy vs.510.23 cGy, relative difference[R]=23.07%).R decreased slowly at t=[-50,-5], increased sharply at t=[-5, 0], and reached the maximum value at a=0.3.After a rapid decrease at a=[0.3, 4], R decreased slowly at a=[4, 50].The correlation analysis of the traditional physical volume dose threshold also showed that the LEUD (at a=0.3) was correlated with V5, V10, V20, and MLD (r=0.929, P<0.05).Conclusions For patients receiving VMAT for thoracic cancer, LEUD (at a=0.3) can distinguish between patients with and without RP.Therefore, LEUD is recommended to be<510 cGy.A combination of LEUD and conventional physical dose has a good clinical predictive value for RP under non-uniform irradiation.

Objective To explore an effective method in reconstructing of burn scar contracture of the chin and neck in children. Methods The blood in the transverse cervical artery might flow reverse into the suprascapular artery and the posterior intercostals perforators through anastomic branches. An axial skin flap in the cervico-scapulo-dorsal area could be adequately supplied by this artery, and we had therefore used this flap to repair burn scar in the neck. Results 30 cases obtained satisfactory clinical results with this flap. The follow-up for two years in 18 patients showed that the neck of the patients moved about freely, and the develpment of the chin was not influenced, and the color of flaps was fine. Conclusion The blood supply of the flap is reliable and stable. So it is a very useful method in reconstructing the burn scar of the chin-neck in children.

Objective To report 11 cases with the tissue defects of their upper or lower limbs repaired with the anastomsed medial sural artery perforator flaps.Methods The free medial sural artery perforator flaps,with length of 8 cm to 15 cm and width of 6 cm to 14 cm were used for tissue defect reconstruction of the distal upper or lower limbs in 11 cases,including 6 females and 5 males.The flap was harvested from the ho- mo-lateral calf,confined between the posterior-medial edge of the tibia and the middle line of the calf and a- bove the distal half part of the medial sural muscle,with a same axis of this muscle.Results Ten cases survived very well,which was relatively thin,and the donor site can be acceptable.One case resulted in a complete flap necrotized and covered with a split skin graft.No obvious motor function defect was observed of the donor leg.Conclusion The anastomsed medial sural artery perforator flap is alternative donor flap for the upper or lower limb tissue defect repair,especially for the defect in the hand or foot.

Aim To investigate the mechanism for the inhibitory effect of hydrogen sulfide on the expression of tissue factor(TF)induced by oxidative low-density lipoprotein(ox-LDL)in endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs ) were treated with 50 mg·L-1 ox-LDL in the absence or presence of different concentrations of NaHS (25 , 50,100 and 200 μmol·L-1 )for 24 h.The mRNA expression and protein content of TF in HUVECs were determined by reverse transcription PCR and ELISA, respectively.The content of intracellular reactive oxy-gen species (ROS)was determined by DCFH,an oxi-dative sensitive fluorescent indicator.The activation of nuclear factor-kappaB (NF-κB)was estimated by its expression in nuclear extracts analyzed by Western blot.Results Ox-LDL induced TF mRNA expression and increased TF protein content in HUVECs.The in-crease in intracellular ROS production and the activa-tion of NF-κB were observed in HUVECs treated with ox-LDL.However,NaHS could markedly inhibit the increases in TF mRNA and protein levels induced by ox-LDL.Also the elevation of intracellular ROS pro-duction and the activation of NF-κB elicited by ox-LDL were significantly suppressed by pretreatment with NaHS.In addition,pretreatment with BAY 1 1-7082 (10 μmol·L-1 ),the inhibitor of NF-κB or N-acetyl-L-cysteine(1 mmol·L-1 ),an antioxidant,could also decrease the TF mRNA and protein level as well as ROS production and NF-κB activation induced by ox-LDL in HUVECs,similar to the effects of 200 μmol· L-1 NaHS.Conclusion The mechanism for the in-hibitory effect of H2 S on the ox-LDL- induced TF ex-pression in endothelial cells may be related to inhibi-ting intracellular ROS production and subsequently NF-κB activation.

BACKGROUND/AIMS: The motility change after peroral endoscopic myotomy (POEM) in achalasia is currently focused on lower esophageal sphincter (LES). This study aims to investigate the correlation of motility response between distal and proximal esophagus after POEM. METHODS: A total of 32 achalasia patients who received POEM and high-resolution manometry (HRM) were included for analysis. Eckardt score was used to assess symptom improvement. HRM was applied for studying motility. Main parameters analyzed were (1) LES: resting pressure (restP), 4-second integrated relaxation pressure; (2) esophageal body (EB): contractile integral of distal segment with myotomy (CI-DM) and proximal segment without myotomy (CI-PNM); and (3) upper esophageal sphincter (UES): relaxation pressure (UES-RP). RESULTS: There were 6 type I, 17 type II, and 9 type III achalasia patients included for analysis. (1) Eckardt score, LES tone, CI-DM, CI-PNM and UES-RP were reduced remarkably after POEM (P < 0.001). (2) no significant correlation was noted between LES tone and contractile intergral of EB. (3) a positive linear correlation of CI-DM and CI-PNM changes was detected (P < 0.001). (4) the change of UES-RP was positively correlated with the change of contractile integral of EB (P < 0.001). CONCLUSIONS: Myotomy of the distal esophagus would attenuate proximal EB contraction and assist UES relaxation in achalasia patients after POEM.
Esophageal Achalasia* ; Esophageal Sphincter, Lower ; Esophageal Sphincter, Upper* ; Esophagus* ; Humans ; Manometry ; Relaxation*

OBJECTIVETo investigate the effect of Lycopene (Lyc) on Ang II induced oxidative stress in H9c2 cell line derived from rat cardiac tissue,and to explore related mechanisms.

METHODSH9c2 cells were divided into 6 groups: control group, Ang II group (1 µmol/L), Ang II (1 µmol/L) + low dose Lyc (3.125 nmol/L) group, Ang II (1 µmol/L) + moderate dose Lyc (6.25 nmol/L) group and Ang II (1 µmol/L) + high dose Lyc (12.5 nmol/L) group and Lyc group (12.5 nnmol/L). Cell growth was determined by CCK8 assay, ROS generation was detected using a Microplate reader and Fluorescence microscopy, the expression of NOX2 was determined by Western blot, mRNA expression of p47(phox), SOD1 and SOD2 were determined by Real Time-PCR, MDA was detected by ELISA kit.

RESULTSCompared to control group,cell survival was significantly reduced and ROS generation was significantly increased post Ang II stimulation,cotreatment with Lyc significantly improved cell survival and reduced ROS generation in a dose-dependent manner (all P < 0.01). mRNA expression of SOD1 and SOD2 was significantly downregulated while MDA concentration was significantly increased in Ang II treated cells, which could be significantly reversed by cotreatment with Lyc in a dose dependent manner (all P < 0.01). Protein expression of NOX2 and mRNA expression of p47(phox) were significantly upregulated post Ang II and which could be significantly downregulated by cotreatment with Lyc in a dose-dependent manner (all P < 0.01).

CONCLUSIONLyc could attenuate Ang II induced oxidative stress and this effect is linked with its capacity of reducing ROS generation and enhancing cellular ROS scavenging ability in H9c2 cells.

Angiotensin II ; Animals ; Carotenoids ; pharmacology ; Cell Line ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Down-Regulation ; Heart ; Oxidative Stress ; RNA, Messenger ; Rats ; Reactive Oxygen Species ; metabolism ; Up-Regulation

OBJECTIVETo examine the urinary level of tissue factor (uTF) and its procoagulant activity (PCA) in patients with diabetes mellitus, and explore the relationship between uTF and renal damage in diabetes mellitus.

METHODSEighty-six patients with type 2 diabetes mellitus were divided into 3 groups according to urine albumin excretion (UACR), namely normal albuminuria group, microalbuminuria group and macroalbuminuria group. The levels of uTF, PCA, blood urea nitrogen (BUN), serum creatinine (CRE), serum cystatin C (CYSC), glycohemoglobin A1c (HbA1c), and high-sensitivity C-reactive protein (hs-CRP) were measured in all the patients and 21 healthy controls.

RESULTSCompared with normal control, the diabetic patients showed significantly increased levels of uTF and PCA. The urinary TF-PCA was positively correlated to BUN, CYSC, CRE, UACR, fasting glucose and hs-CRP, but not to uTF; only hs-CRP, UACR were positively correlated to uTF.

CONCLUSIONuTF is probably implicated in the development and progression of diabetic nephropathy.

Adult ; Albuminuria ; urine ; Blood Coagulation ; Case-Control Studies ; Creatinine ; urine ; Diabetes Mellitus, Type 2 ; physiopathology ; urine ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; urine

OBJECTIVETo explore the correlation between the biological characteristics of volunteer donors and sperm parameters.

METHODSSperm parameters were analyzed for 778 primary volunteer donors by CASA system with standard methods recommended by WHO including sperm concentration, motility rate, VCL, VSL, ALH, LIN, STR and BCF, and their biological characteristics were recorded.

RESULTSOf all the volunteer donors, students accounted for more than a third, 89.2 percent were under the age of 35 years, 71.6 percent between 166 and 175 cm in height, 91.7 percent of normal weight and 77.1 percent with college education. The median sperm concentration and motility rate were 107.00 x 10(6)/ml and 75 percent respectively. Age was weakly correlated with such semen indexes as concentration (r = 0.210, P = 0.000) , motility rate (r = 0.213, P = 0.000), volume (r = 0.165, P = 0.002), VAP (r = 0.259, P = 0.000), VSL (r = 0.281, P = 0.000), VCL (r = 0.190, P = 0.000), BCF (r = 0.243, P = 0.000) and LIN (r = 0.192, P = 0.000). The semen indexes of the 26-35 age group were a little better than those of the 20-25. Height showed no correlation to semen indexes. Education and occupation were somewhat related with other sperm parameters (P < 0.01) than ALH (P = 0.695/0.886).

CONCLUSIONAge, occupation and education bear some correlation with sperm parameters except ALH, while height has none with any of them.

Adult ; Age Factors ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Occupations ; Semen ; cytology ; Sperm Count ; Sperm Motility ; physiology ; Spermatozoa ; physiology ; Students ; Tissue Donors ; statistics & numerical data

OBJECTIVESTo identify whether Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP1) can induce tumor necrosis factor receptor-associated factor 1 (TRAF1) expression and promote its anti-apoptosis activity via the NF-kappaB signaling pathway, and assess that LMP1 suppresses apoptosis in nasopharyngeal carcinoma (NPC).

METHODSA stable transfected cell line HNE2-LMP1 was established by introducing LMP1 cDNA into HNE2 cells. Transactivation of TRAF1 was determined by luciferase reporter assay, while expression of TRAF1 mRNA was detected by RT-PCR and expression of TRAF1 protein and caspase 3 by Western blot analysis. Apoptosis activity was observed through fluorescence staining.

RESULTSLMP1 induced TRAF1 expression in NPC cells and caused a decrease in apoptosis. This induction could be blocked by antisense LMP1. Moreover, LMP1-mediated induction of a TRAF1 promoter-driven reporter gene was significantly impaired when the kappaB site kappaB1 or kappaB5 was disrupted, whereas mutation of kappaB3 had only a minor effect on LMP1 dependent up-regulation of the reporter gene.

CONCLUSIONLMP1 induces TRAF1 expression and promotes its anti-apoptosis activity via the NF-kappaB signaling pathway, which may be one of the mechanisms that LMP1 uses to suppress apoptosis in NPC cells.

Apoptosis ; physiology ; Humans ; NF-kappa B ; physiology ; Nasopharyngeal Neoplasms ; physiopathology ; Protein Biosynthesis ; Signal Transduction ; physiology ; TNF Receptor-Associated Factor 1 ; Tumor Cells, Cultured ; Viral Matrix Proteins ; physiology