Objective To develop a method for the determination of acrylamide in flocculant by gas chromatography-mass spectrometry.Methods Acrylamide was derived through bromination and extracted by organic solvent.GC-MS method was used to identify the target substance and quantify acrylamide.Results The recoveries obtained from samples spiked with standards were in the range of 70.2%-109.2% and relative standard deviations (n=5) was less than 5.0%.Linearity of peak area was obtained over a wide range of concentration (4.3-2 000 ?g/ml),and the correlation coefficients was 0.999 4.Conclusion The method is reliable,selective and reproducible and it is applicable to the determination of the acrylamide in flocculant.

Toll-like receptor 5 (TLR5) is one of the pattern recognition receptors and recognizes the flagellin protein of bacteria.It activates innate immune responses and induces production of a series of cytokines.TLR5 functions as a bridge linking innate and adaptive immunities.It is known that TLR5 plays an important role in the occurrence and development of certain infectious diseases.This review summarizes the relationships of TLR5 polymorphisms with the development of infectious diseases and discusses the possible pathogenesis.

OBJECTIVE To explore the rational antibiotics usage in hepato-biliary department.METHODS The antibiotics used in liver surgery department were investigated and evaluated according to the standards.RESULTS Totally 235 patients (88.01%) among 267 patients from 2005 to 2007 were used antibiotics,of which 18.30% for therapy,81.70% for prophylaxis and from them 83.40% were for systemic use. The specimens from 35 patients among 235 patients with liver disease,were sent to do antibiotic susceptibility test. Most of the patiants were given therapy by experience.CONCLUSIONS The administration department in the hospital should enhance its work by forming institutions and standards,and inspecting regularly to guarantee the safety of antibiotics usage.

Objective: To investigate the immunosuppressive effect of sinomenine (SIN) on xenogenic skin transplantation in mice.Methods: BALB/c→C57BL/6 tergal skin transplantation model was established by an operation, and then the mice were divided into 5 groups at random, namely sham group, model group, SIN group (30 mg·kg-1), ciclosporin A group (CsA) (10 mg·kg-1) and combination of SIN and CsA group (SIN 30 mg·kg-1 and CsA 5 mg·kg-1) (n=10).All the mice were intraperitoneally administered once a day for ten days.The survival days of skin graft were recorded, and the IL-2 levels in plasma were determined by ELISA respectively on the 4th and 8th day after the operation.Results: The mean survival days of skin graft in the groups treated with different drugs were significantly prolonged when compared with that in the model group (P<0.01).The combination of SIN and CsA administration showed longer mean survival days than SIN or CsA (P<0.05 or P<0.01).The IL-2 levels in plasma in the groups treated with different drugs were significantly reduced than those in the model group on the 4th and 8th day after the operation (P<0.01).Conclusion: SIN may have a good immunosuppressive effect in the mice with xenogenic skin transplantation, and the combination of SIN and CsA shows a synergistic effect.

Objective:To investigate the effects of sodium aescinate on endothelin ( ET ) and calcitonin gene -related peptide ( CGRP) contents in the gerbils with cerebral ischemia/reperfusion injury ( CI/R) . Methods:The gerbil model of CI/R was prepared by bilateral common carotid arteries ligation for 10 min followed by 2-hour reperfusion. Sodium aescinate (i. p. , 10, 20 and 40 mg· kg-1 ) was administered once a day for 3 days before the operation and once every 1 h after the operation, respectively. The levels of ET and CGRP in brain tissue homogenate were determined by a radioimmunoassay method. Results:Sodium aescinate significantly in-hibited the level of ET (28. 69-37. 03 ng·L-1) in the brain tissue of gerbils with CI/R compared with the model group (P<0. 05 or P<0. 01), while showed no evident influence on CGRP (P>0. 05). Conclusion:Sodium aescinate has obvious protective effects a-gainst CI/R in gerbils, which may be due to its inhibitory action on ET levels in brain tissue.

Objective:To investigate the protective effects of paeoniflorin on myocardial ischemia/reperfusion ( MI/R) injury in rats. Methods:The rat model of MI/R injury was prepared by coronary artery ligation for 30 min followed by 2-hour reperfusion. Then the rats were randomly divided into 5 groups:sham group, model group, paeoniflorin group respectively at high, medium and low dose (20, 10 and 5 mg·kg-1, n=10). Paeoniflorin was respectively injected via tail vein 1 h before the operation and at the beginning of the reperfusion. The CK, LDH and SOD activities and MDA contents in serum of rats were examined, and the area of myocardial in-farction was also calculated after the reperfusion. Results:Paeoniflorin (20 and 10 mg·kg-1 ) could significantly reduce the area of myocardial infarction when compared with the model group (P<0. 05). Paeoniflorin could obviously inhibit the CK and LDH activities (P<0. 05 or P<0. 01) and enhance the SOD activity (P<0. 01) in serum of MI/R rats. The MDA content in paeoniflorin groups at high and medium dose was significantly lower than that in the model group (P<0. 05). Conclusion:Paeoniflorin pretreatment shows effectiveness against MI/R injury, which may be associated with the inhibition of lipid peroxidation.

AIM: To evaluate the influence of radiofrequency (RF) hyperthermia on immunity function in mice. METHODS: The expression pattern of T helper type 1 (Th1) and T helper type 2 (Th2) cytokines in splenocyte culture supernatants, mainly the expression levels of IFN-?, IL-2, IL-4 and IL-10 in splenocyte culture supernatants of mice in tumor-bearing group, surgical resection group, RF therapy group and normal control group were detected by enzyme-linked immunoadsordent assay (ELISA). RESULTS: IL-2 concentration in two weeks after RF therapy group was higher than that in two weeks after surgical resection and normal control groups (P0.05). CONCLUSION: RF hyperthermia may activate the transformation from Th2 to Th1 and facilitate the excretion of Th1 type cytokines that play an important role in the anti-tumor immunity.

OBJECTIVE To analyze antibiotic usage of inpatients in an internal medicine department,probe into the features of antibiotic usage and provide rference for the rational clinical drug-use.METHODS We selected randomly 3102 samples from inpatients in our Internal Medicine Department during the period from Jan to Jun,2007,and analyzed the situation of the antibiotic application.RESULTS The percentage of antibiotic application was 66.18% among 3102 patients,totally 28 kinds from 10 species of antibiotics were used.Cephalosporins and quinolones were mostly applied,and more often they were used in combination.CONCLUSIONS Medication frequency of cephalosporins and quinolones ranks the first in the DDDs,and remains as the primary choice of anti-infectives used.

BACKGROUND:Extraction methods of human amniotic mesenchymal stem cells are inconsistent in the number of cells.
OBJECTIVE:To explore the optimal method to in vitro isolate and culture human amniotic mesenchymal stem cells.
METHODS:Under sterile conditions, ful-term cesarean fetal amniotic membrane was cut into pieces, then to isolate human amniotic mesenchymal stem cells by seven methods in four experiments. In experiment 1, human amniotic mesenchymal stem cells were isolated by the fol owing three methods:(1) 0.05 g/L trypsin digestion for 10 minutes fol owed by 0.75 g/L col agenase digestion for 60 minutes;(2) 0.75 g/L col agenase I for 120 minutes;(3) co-digestion with 0.05 g/L trypsin and 0.75 g/L col agenase for 60 minutes. In experiment 2, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes fol owed by 0.75 g/L col agenase digestion for 30 minutes. In experiment 3, the samples were digested by two methods:(1) 0.05 g/L trypsin digestion for 30 minutes×2, fol owed by 0.75 g/L col agenase digestion for 60 minutes;(2) 0.05 g/L trypsin digestion for 40 minutes×2, fol owed by 0.75 g/L col agenase digestion for 60 minutes. In experiment 4, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes×2, fol owed by 1 g/L col agenase digestion for 60 minutes. Fol owing morphology observation under a microscope, we studied the most suitable method for isolating human amniotic mesenchymal stem cells.
RESULTS AND CONCLUSION:Digestion with 0.05 g/L trypsin for 30 minutes twice fol owed by 1 g/L of col agenase digestion of 60 minutes was the most suitable isolation and culture condition in vitro. cells became elongated fusiform or star-shaped with rich cytoplasm, and nuclei were round with 1-3 nuts. We can harvest the most number of human amniotic mesenchymal stem cells using the method described in experiment 4.

AIM:To observe the effect on NF-?B pathway and cell motility in breast cancer cell lines after transfection of dominant negative I?B? plasmid.METHODS:After stable transfection of mutant I?B? plasmid into highly metastatic breast cancer lines MDA-MB-231 and MDA-MB-435,we detected NF-?B binding activity by EMSA,cell growth ability by cell growth curve,colony forming test,and cell motility by millicell-PCF chamber.RESULTS:Constitutive activities in MDA-MB-231 and MDA-MB-435 were observed.Stable transfection of a dominant negative Ⅰ?B? resulted in downregulation of NF-?B binding activity,thus inhibited cell mobility without significant effect on cell growth.CONCLUSION:Cell migration ability is inhibited in highly invasive breast cancer cells by inhibition of NF-?B pathway in vitro.