OBJECTIVE:To study the interaction of cefprozil (CE) with bovine serum albumine (BSA). METHODS:Under the temperatures of 289,299 and 309 K,the interaction of CE with BSA for 50 min had been studied by fluorescence quenching, UV spectrometry and synchronous fluorescence spectroscopy. Quenching constant(KSV)and speed constant(Kq)were calculated by Stern-Volmer equation. Static quenching constant(KLB)was obtained by Lineweaver-Burk equation,and UV spectrogram was used to determine the type of quenching. Double logarithmic equation was used to calculate the binding constants (Kb) and the number of binding site(n). Thermodynamic equation was used to obtain ΔH,ΔS,ΔG. Hill's coefficients(nH)was obtained by Hill equa-tion. RESULTS:At three different temperatures,with CE concentration increasing,fluorescence intensity of BSA decreased regular-ly. The value of KSV,Kq,KLB,Kb and n and nH decreased with the temperature increasing. ΔH,ΔS and ΔG were lower than 0. The numbers of binding sites were approximately equal to 1 and nH<1. CONCLUSIONS:CE statically quench the fluorescence of BSA,and the binding of them have been found to certain extent. The process of binding is spontaneous exothermic process. The main binding forces include Hydrogen bonds and Van der Waals forces,and primary binding site for CE is located at sub-domain ⅡA of BSA. There was some negative cooperative effect. CE would not affect the conformation of BSA. The binding site of CE and BSA is near by tyrosine residue.

Obiective To explore the interaction of cefonicid sodium(CS) with bovine serum albumin(BSA) by fluorescence and absorption spectroscopy. Methods The rate constant(Kq), quenching constant(Ksv), static fluo-rescence quenching association constant(KLB),binding site number(n) and binding constant(Kb) were calculated using Stern-Volmer, Lineweaver-Burk and double logarithm equations. Results CS was able to bind to BSA. The probable quenching mechanism of BSA by CS was mainly static quenching due to the formation of a CS-BSA com-plex. The results of thermodynamic parameters indicated that electrostatic force plays the main role in the binding process and the binding process was spontaneous. There was a single class of binding site for the BSA with CS. The primary binding site for CS was located at sub-domainⅡA of BSA and near by tyrosine residue. There was almost some negative cooperative effect. The results obtained from synchronous fluorescence showed that CS could change the microenvironment of Tyrand Trp residues of BSA. Conclusion The interaction between CS and BSA is dynam-ic. There is a single class of binding site for the BSA with CS. The obtained results provide references for its clini-cal application.

Objective To observe the clinical efficacy of acupoint injection plus nerve electrical stimulation in treating deglutition disorders after cerebral stroke. Method Seventy-seven patients with deglutition disorders after cerebral stroke were randomized into an electrical stimulation group of 24 cases, a hydroacupuncture group of 26 cases, and a comprehensive group of 27 cases, to respectively receive Vitalstim electrical stimulation, acupoint injection of Mecobalamin, and both of the treatments, 5 d as a treatment course, with 2-day interval between two courses, for 4 courses in total. The modified water-drinking test and Standardized Swallowing Assessment (SSA) were adopted to evaluate the therapeutic efficacy before the intervention, after 20-day treatment, and after 60-day treatment, and the therapeutic efficacies were compared. Result The modified water-drinking test and SSA scores were significantly changed in the three groups after 20-day treatment compared with that before the intervention, and the scores in the comprehensive group were superior to that of the electrical stimulation group and hydroacupuncture group. However, on the 60th day, the scores were equivalent among the three groups. Conclusion The Vitalstim electrical stimulation and acupoint injection of Mecobalamin both can produce certain treatment effects for deglutition disorders after cerebral stroke;the two methods can work in a synergistic way and can boost the improvement of swallowing function.

The interaction betweenYan-Hu-Ning (YHN) and bovine serum albumin (BSA) was investigated in order to provide further theoretical evidences on action mechanism study between YHN and proteins within the organism. Under optimal conditions, the interaction between YHN and BSA was studied by fluorescence quenching, ultraviolet absorption spectrometry and synchronous fluorescence spectroscopy. At the temperature of 283.15 K, 298.15 K and 313.15 K, quenching constant (KSV) and speed constant (Kq) were calculated by S-V curves. Static quenching constant (KLB) was obtained by L-B double reciprocal equation. Double logarithmic equation was used to calculate the binding constants (Kb) and the number of binding site (n). Thermodynamic equation was used to obtainΔH,ΔS,ΔG. Hill’s coefficients (nH) was obtained by Hill equation. The results showed that at three different temperatures, along with the increasing of YHN concentration, the fluorescence intensity of BSA decreased regularly. The value of KSV, Kq, KLB, Kb, n and nH decreased with the increasing of temperature;ΔG < 0,ΔH < 0,ΔS < 0; n was approximately equal to 1; nH > 1. It was concluded that YHN-BSA complex quenched the intrinsic fluorescence of BSA. The mechanism of fluorescence quenching was static quenching. The main binding forces were deduced as hydrogen bonds and Van der Waals forces from calculated values of thermodynamic parameters. YHN and BSA can form a binding site, which indicated certain binding interaction between YHN and BSA. YHN can be stored and transported by protein within the body. Free energy was produced to transformΔG into negative value. It showed that the process of binding between YHN and BSA was spontaneous. The nH was more than 1. It indicated that YHN had positive cooperative effect. The primary binding site was located at subdomainⅡA. The synchronous fluorescence spectra showed certain influence on the conformation of BSA by YHN. It led to the weakening of polarity within BSA and the binding site to be closer to the tyrosine.

ObjectiveTo investigate the effect of XPG down-regulation gene expression towards the proliferation of epithelial ovarian cancer cells and its chemosensitivity to platinum.Methods The small interference RNA ( siRNA ) -XPG fragments were designed and tranfected into SKOV3/DDP cell lines by lipofectamine transiently for choosing the best siRNA-XPG fragment to silence XPG gene expression.The pGPU6/GFP/Neo vector was used to construct the siRNA-XPG vectors,which was transfected into SKOV3/DDP cell line with expression of XPG gene.Real-time PCR and western blot were employed to confirm the silencing efficacy of siRNA-XPG.The growth curve of cells,cell cycle,the drug-resistance index of cells and intracellular drug concentration were measured by 4-methyl-thiazolyl-tetrazolium (MTT),flow cytometer (FCM) and high performance liguid chromatograph respectively.Results( 1 ) Real-time PCR results showed that XPG mRNA expression copy number in SKOV3/DDP tranfected with siRNA-XPG-733 fragment was 1.050 ± 0.023,which was significantly lower than that in SKOV3/DDP tranfected with other siRNA-XPG fragments(P ＜ 0.05,respectively),and was chosed to construct the siRNA-XPG vectors.The XPG mRNA expression was down-regulated in short hairpin RNA (shRNA)-XPG-733-SKOV3/DDP cell lines that confirmed by western blot.( 2 ) The growth curve showed that growth velocity of shRNA-XPG-733-SKOV3/DDP cell lines was lower than that of shRNA-GAPDH and shRNA-NC cell lines( P ＜ 0.05,respectively).The results of FCM also showed that 34.0％ of cells in shRNA-XPG-733-SKOV3/DDP cell lines were in S + G2/M phase,while only 58.7％ and 51.3％ in shRNA-GAPDH and shRNA-NC cell lines respectively ( P ＜ 0.05,respectively).( 3 ) The drug-resistance index of shRNA-XPG-733-SKOV3/DDP cell lines [ 50％ inhibiting concertration( IC50 ):( 13.79 ± 0.06) μg/ml ] was lower than that in shRNA-GAPDH and shRNANC cell lines [ IC50:( 27.84 ± 0.34 ) μg/ml and ( 28.32 ± 0.42 ) μg/ml,respectively ] statistically significant (P ＜ 0.05,respectively) ; but there was not statistically significant difference in intracellular drug concentration between shRNA-XPG-733-SKOV3/DDP cell lines [ (0.026 ± 0.005 ) μg/ml ] and shRNAGAPDH [ (0.024 ± 0.003 ) μg/ml ] and shRNA-NC cell lines [ ( 0.025 ± 0.007 ) μg/ml ] after treated by cisplatin in vitro ( P ＞ 0.05,respectively ).Conclusion The down-regulating of XPG gene resulted in slowing growth velocity and descending the drug-resistance index of shRNA-XPG-733-SKOV3/DDP cell lines,which may be related with descending in capability of DNA excision repair in cells.

Objective To compare anatomic resection (AR) and non-anatomic resection (NAR)for hepatocellular carcinoma (HCC) as a factor in preventing intra-hepatic recurrence and local recurrence after the initial surgical procedure.Methods A systematic review and Meta-analysis of nonrandomized trials comparing anatomic resection with non-anatomic resection for HCC published from 1990to 2010 in PubMed and Medline,Coehrane Library,Embase,and Science Citation Index were searched.Intra-hepatic recurrence,including early and late recurrence,and local recurrence were primary outcomes.5-year survival and 5-year disease-free survival were secondary outcomes.Pooled effect was calculated by utilizing either fixed effects model or random effects model.Result Eleven nonrandomized studies including 1576 patients were identified and analyzed.810 patients were in the AR group and 766 were in the NAR group.Patients in the AR group were characterized by lower prevalence of cirrhosis,more favorable hepatic function,and larger tumor size and higher prevalence of macrovascular invasion compared with patients in the NAR group.Anatomic resection significantly reduced the risks of local recurrence (OR,0.27; 95％ CI,-0.17～0.43; P＜0.001) and achieved a better 5-year disease-free survival (OR,2.10; 95％ CI,-1.41 ～3.12; P=0.001) in HCC patients.Also,anatomic resection was marginally effective in decreasing early intra-hepatic recurrence.However,anatomic resection was not advantageous in preventing late intra-hepatic recurrence.No significant differences were found between the AR and NAR groups with respect to postoperative morbidity,mortality,and length of hospitalization.Conclusion Anatomic resection was recommended to be superior to non-anatomic resection in reducing the risks of local recurrence,early intra-hepatic recurrence and achieving a better 5-year disease-free survival in HCC patients.

Hepatocellular carcinoma is one of the most common malignancies in the worldwide.Selected patients with hepatocellular carcinoma are candidates for curative resection,but nevertheless there is a high risk of tumor recurrence.Microvascular invasion (MVI) is an aggressive biological behavior and has repeatedly been identified as a risk factor for prognosis after curative treatment,meanwhile,it is now becoming increasingly concerned.It would be of great significance to distinguish MVI in an early stage and choose an appropriate treatment timely to get a definite improvement for the long-term survival in patients with hepatocellular carcinoma after curative treatment.This review focuses on some certain issues of MVI.

Objective:To clone and express the Der f 7 recombinant protein from Dermatophagoides farinae and prepare the Der f 7 monoclonal antibody.Methods: The Der f 7 recombinant protein was expressed in prokaryotic expression system of pET28a(+)-Der f 7.BALB /c mice were immunized with the recombinant protein.Myeloma cells and spleen cells were fused,and hybridoma cells were screened by ELISA.Hybridoma cells were injected into the mice abdominal cavity to obtain ascites.It was purified by protein A agarose medium ascites,and then to dentified the titer and purity of the antibody.The specificity of the antibody was identified by Western blot.Results: Three hybridoma cells which stably secret recombinant Der f 7 monoclonal antibody were obtained.The monoclonal antibody had high purity,the titer was higher than 1∶243 000.Western blot showed that Der f 7 recombinant protein could be recognized well.Conclusion: We successfully obtained Der f 7 monoclonal antibody,which can be used for the quantification and localization of Der f 7 allergen and the diagnosis and treatment of allergic diseases.

Objective To develop a fluorescence signal activatable multifunctional molecular probe with theranostics function through combining ultrasmall gold nanorods(UGNRs) with fluorescein,and to evaluate its therapeutic effect on photothermal therapy (PTT) in breast cancer cells.Methods The UGNRs were synthesized by the one-pot seedless method,then the functionalized modification of UGNRs were conducted using cysteamine.Finally,the activatable ultrasmall gold nanorods (AUGNRs) were synthesized by amide condensation of —NH2 of cysteamine and —COOH of carboxylated fluorescein CyS.The cell uptake ability and GSH-mediated imaging ability of AUGNRs were studied using breast cancer 4T1 cells.4T1 cells co-cultured with AUGNRs were irradiated with 808 nm excitation light,and the PTT effects were assessed by MTT colorimetric staining and calcein-AM/PI staining.Results The AUGNRs were synthesized successfully,which could be uptaken by 4T1 cells quickly and efficiently,and could achieve intracellular glutathione (GSH) triggered fluorescence recovery.No obvious cytotoxicity of AUGNRs to 4T1 cells was observed in the co-cultivation.Moreover,obvious PTT effects could be induced by 808 nm laser,which could effectively kill 4T1 cancer cells.Conclusion The fluorescence signals of AUGNRs can be induced by intracellular GSH,and tumor cell destruction can be achieved by 808 laser-excitated PTT effects.

Objective To investigate the change and clinical significance of serum hepcidin,serum ferritin (SF),transferrin re-ceptor (sTfR)and serum iron (SI)in patients in type 2 diabetes(T2DM).Methods 130 patients with T2DM were divided into 2 groups according to the 24 hour urine microalbumin (mAlb)quantitative:group A for trace microalbumin group 45 ca-ses (mAlb30~300 mg/24 h),group B for normal albuminuria group of 85 cases,an alternate period of 45 cases of healthy physical examination for group C (control group).Results Serum hepcidin and SF of group A (42.27±32.12 ng/ml,211.6 ±107.2 ng/ml)were significantly higher than those in group B (26.12 ± 18.36 ng/ml,179.1 ± 109.7 ng/ml)and the healthy control group (P <0.05),hepcidin and SF of group B was significantly higher than that of the control group (9.47 ±1.65 ng/ml,84.41±47.10 ng/ml,P <0.01),SI and transferrin receptor(sTfR)has no statistical significance between the three groups (P >0.05).Correlation analysis showed that patients with type 2 diabetes hepcidin was positively related with SF (P <0.05),hepcidin and sTfR,SI had no significant correlation.Conclusion These results indicated that there existed serum hepcidin and SF increased iron overload and iron metabolism disorder in type 2 diabetes.Therefore,detection of serum iron and SF can be used as a predictor of diabetes early renal damage.