Objective To detect the expression of SSX and to correlate it with clinical indicators of primary hepatocellular carcinoma (HCC).Methods The expression of SSX1-5 mRNA and SSX1 protein were respectively detected by RT-PCR and Western blot and immunohistochemistry staining.The relation between the expression of SSX mRNA and SSX1 protein with clinical indicators were analysed.Results SSX1,SSX2,and SSX3 mRNA were expressed in hepatocellular carcinoma cell lines BEL-7404,Hep G2,and SMMC-7721.In 26 HCC samples,SSX1-SSX5 mRNA was detectable in 53.8％,42.3％,50.0％,46.2％ and 26.9％.The expression of SSX1 mRNA was not related to serum AFP levels (P ＞0.05).Specific expression was both found in the normal group and the high value group.The expression rate of SSX1 mRNA was 85.7％ in the older group,which was higher than in the younger group (16.7％,P ＜ 0.05).The expression rate of SSX1 protein was 50％ in HCC tissues,which was not seen in the caner-adjacent or cirrhosis tissues.In 49 HCC paraffin tissue section samples,the expression rate of SSX1 protein was higher than that in caner-adjacent tissues (46.9％ vs 18.4％,P ＜ 0.05).The expression rate of SSX1 protein was 68.3％ in the large hepatocellular carcinoma group,which was higher than in the small hepatocellular carcinoma group (29.6％),(P ＜ 0.05).Conclusions SSX1 mRNA is expressed with a high percentage and specificity in HCC and their products are new potential promising targets for antigen-specific immunotherapy of HCC.The detection of SSX1 expression has the potential value for auxiliary diagnosis of HCC.

Objective To develop a precise and reliable method to determine mammalian cell survival using the triphe-nyltetrazolium chloride (TTC) solution.Methods Different amounts of TTC solution were added to the cultured cells in order to find the optimal concentration .The correlation between the cell number and the measured optic density value was analyzed.In addition, the TTC method was compared with traditional MTT assay .Results The TTC assay could precisely reveal the viability of cultured cells as well as the MTT method , but the non-specific reaction of the MTT reagent by other antioxidants could result in unreliable overestimation of the cell number .Conclusion The TTC assay can be used to deter-mine the survival of cultured cells more precisely .

Objective Sodium-bile acid cotransporter plays an important role in the development of Echinococcus.The present study aimed to clone sodium bile acid cotransporter gene in Echinococcus multilocularis (EmSBACT5) and to analyze the bioinformatics of its coding protein.Methods EmSBACT5 gene was amplified by reverse transcription RCR (RT-PCR) technology and its nucleotide sequence was sequenced.Bioinformatics softwares were used to predict and analyze the physical and chemical properties, hydrophilicity/hydrophobicity, transmembrane domain, post-translational modification sites, structural domain, secondary structure, tertiary structure, subcellular localization and biological functions of the coding protein.Results The complete open reading frame was amplified with 654 bp in length, encoding 217 amino acids.The homology of the nucleic acid sequence and amino acid sequence of EmSBACT5 gene were 98% and 96% with the published SBACT5 in Echinococcus granulosus (EgSBACT5) respectively.Protein analysis results showed that the molecular formula of EmSBACT5 protein was C1141H1797N273O284S11.Its relative molecular mass was 24240 and isoelectric point was 8.99.There were 9 post-translational modification sites and 4 typical domains.Alpha helical, β-sheet, β-turn and random coil accounted for 29.95%, 31.80%, 7.83% and 30.41%, respectively.This protein was a hydrophobic membrane protein and was mainly located in the cytoplasmic membrane, and it might play a role in the processes of material transport and signal transmission.Conclusion The EmSBACT5 gene was cloned successfully and the informatics characteristics of its coding protein were obtained, which provides basic information for prevention and control of echinococcosis.

At present,corresponding cell-free parasite DNA molecules (CFPD) has been detected in serum,plasma,urine,saliva and other bodily fluids of a variety of the patients with parasitic diseases.Due to its high specificity and sensitivity,the CFPD shows a strong advantage of noninvasive diagnosis and continuous monitoring,etc.in parasitic diseases.This article namely reviews the current research of CFPD in the patients with parasitic disease at home and abroad in recent years,so as to provide new ideas for the development direction of parasitic disease diagnosis in the future.The current related problems are discussed in the mean time.

ObjectiveTo investigate the relationship between tumor-infiltrating mast cell (TIM) and the clinicopathological and prognostic factors of patients with colorectal cancer.MethodsA total of 282 cases of paraffin-embedded colorectal cancer specimens were obtained from the First Affiliated Hospital of Sun Yat-sen University from January 2002 to December 2005.The density of TIM was determined by immunohistochemical staining.According to the mean TIM density detected [ ( 8.4 + 6.5 )/HPF ],all the patients were divided into low-TIM density group (mean TIM density ＜8.4/HPF) and high-TIM density group (mean TIM density ＞8.4/HPF).The clinicopathological factors and the prognosis of patients between high-TIM density group and low-TIM density group were compared.All data were analyzed using the t test or chi-square test.The survival curve was drawn using the Kaplan-Meier method,and the survival of the patients was analyzed by the Log-rank test.The clinicopathological factors were analyzed retrospectively with the univariate and multivariate COX regression model.ResultsTIM was detected in all the patients with colorectal cancer.Significant differences were observed in the number of patients in N stage and TNM stage between patients in the high-TIM density group and those in the lowTIM density group (x2 =6.025,7.410,P ＜ 0.05 ).All patients were followed up till September 2010,the 5-year overall and tumor-free survival rates of patients were 82.9％ and 63.1％ in the low-TIM density group,79.0％ and 59.3％ in the high-TIM density group,with significant difference between the 2 groups (P ＜ 0.05 ).COX proportional hazard regression model revealed that high density of TIM was associated with short overall survival time and tumor-free survival time of colorectal cancer patients ( RR =2.119,95 ％ CI 1.326- 3.386; RR =2.084,95 ％ CI 1.357-3.199,P ＜0.05).The resuhs of multivariate analysis showed that high density of TIM was the independent factor influencing the overall survival time and tumor-free survival time (RR =1.651,95％ CI 1.009-2.702; RR =1.680,95％ CI 1.074-2.629,P ＜ 0.05 ).ConclusionHigh density of TIM is correlated with the N stage and TNM stage of colorectal cancer,and it is an independent predictor of poor survival for patients with colorectal cancer.

Objective To analyze the clinicopathological factors on the prognosis and investigate the necessity of adjuvant chemotherapy for patients with stage Ⅱ colorectal cancer.Methods The clinical data of 255 patients with stage Ⅱ colorectal cancer who were admitted to the First Affiliated Hospital of Sun Yat-Sen University from January 2000 to December 2005 were collected.The survival curve was drawn by Kaplan-Meier method,and the survival rate of the patients were analyzed by Log-rank test.Factors influencing the survival were analyzed by Cox regression model.Results All patients were followed up till April 23,2010,and the mean time of follow-up was (63 ± 22)months.The median survival time was 63 months.The 5-year and tumor-free survival rates were 85.3％ and 83.7％,respectively.The 5-year overall and tumor-free survival rates of patients without preoperative bowel obstruction or perforation were 86.9％ and 85.6％,which were sigaificantly higher than 72.7％and 68.4％ of patients with preoperative bowel obstruction or perforation(x2 =4.546,4.573,P ＜ 0.05 ).The 5-year overall and tumor-free survival rates of patients with negative resection margin were 85.5％ and 83.9％,which were significantly higher than 75.0％ and 75.0％ of patients with positive resection margin(x2 =7.020,6.009,P ＜ 0.05 ).The result of multivariate analysis revealed that preoperative bowel obstruction or perforation were the independent risk factors for patients with stage Ⅱ colorectal cancer(Wald =4.477,relative risk =2.371,95 ％ confidence interval:1.066-5.275,P ＜ 0.05 ).The 5-year overall and tumor-free survival rates were 87.3％ and 86.0％ for patients who received adjuvant chemotherapy,and were 82.2％ and 80.3％ for patients who did not receive adjuvant chemotherapy (P ＞ 0.05 ).Conclusions Preoperative bowel obstruction or perforation are independent risk factors for the survival of patients with stage Ⅱ colorectal cancer.Adjuvant chemotherapy could not improve the prognosis of patients with stage Ⅱ colorectal cancer.

Moxibustion,which is simple for manipulation and effective,has always been a very important way for external treatment.Acupuncture and moxibustion were complementary to each other sincc ancient times.but modem people usually only pay attention to acupuncture and neglect the usage of moxibustion.In this paper the importance of moxibustion is expounded briefly.

Objective To obtain the economic data in the aspects of cost-effectiveness and cost-utility of neonatal congenital hypothyroidism(CH) screening in Foshan City and to make assessment on the effect of screening works.Methods The economic related data were collected by the screening center,literatures,official resources and questionnaire investigation.The statistical tool and Tree Age Pro health decision analysis software were used to conduct the assessment on the cost-effectiveness and cost-utility of screening works.Results The screening program invested 17.95 million Yuan in total during 2000-2007,and the gain benefits was 117.69 million Yuan,net benefit was 9 975.52 ten thousand Yuan.The cost-benefit ratio was 1.00∶6.56.Each investing 3 216 Yuan could avoid a disability-adjusted life year.Conclusion The CH screening item has good economic applicability and is worth investing more funding for further promotion and popularization.

Fluorescence spectroscopy was used to investigate the efficiency of coupling peptides to gold nanoparticles via 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS).The experiment conditions including buffer solution, pH value and concentrations of buffer solution, concentrations of NHS and EDC, concentration ratios of NHS to EDC, and coupling reaction time on the coupling efficiency were investigated and optimized.The experimental results indicated that the optimized experimental conditions were as follows: 25 mmol/L HEPES buffer solution, pH 7.0;2∶1 of concentration ratio of NHS to EDC, 0.4 mol/L NHS, 0.2 mol/L EDC, and coupling reaction time of 24 h.This study may provide references for the relative research involving coupling peptide or protein with gold nanoparticles

Objective Through the detection of the levels of serum betatrophin,anthropometric and biochemical indices,to determine the alteration of betatrophin levels in patients with polycystic ovary syndrome (PCOS) and the relationship between betatrophin levels and metabolic indexes such as 25-hydroxy vitamin D3 [25 (OH) D3].Methods The study group was composed of 24 women with PCOS and 20 age-natched healthy women as controls.Following general physical examination of the subjects,anthropometrie measurements were performed (height,weight,waist circumference,hip circumference).Body mass index (BMI) and waist-hip ratio were calculated;25 (OH) D3,Ca2+,fasting blood glucose,fasting insulin,triglycerides,high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C) were also measured and homeostasis model assessment of insulin resistance (HOMA-IR) were evaluated in all subjects.Serum betatrophin levels were examined with an enzyme-linked immunosorbent assay (ELISA).Using independent sample T test to compare the differences between groups,the linear correlation analysis was performed to study the correlation of betatrophin with anthropometric and biochemical indices,the influencing factors of betatrophin were analyzed by multiple linear regression.Results Circulating betatrophin levels [(0.341±0.034) ng/ml vs.(0.810±0.162) ng/ml,t=4.271,P＜0.001] and HDL-C [(1.014±0.321) mmol/L vs.(1.419±0.287) mmol/L,t=2.218,P＜0.05] significantly decreased in women with PCOS compared with controls,and BMI [(27.691±4.392) kg/m2 vs.(23.310±4.781) kg/m2,t=-2.073,P＜0.05],fasting blood glucose [(5.950±0.411) mmol/L vs.(4.883±0.314) mmol/L,t=-2.142,P＜ 0.05],HOMA-IR [(4.946 ± 0.741) vs.(3.387± 0.397),t=-2.493,P＜0.05],triglycerides [(1.510± 0.848) mmol/L vs.(1.037±0.402) mmol/L,t=-2.223,P＜0.05],LDL-C [(3.431±0.479) mmol/L vs.(2.396±0.435) mmol/L,t=-2.433,P＜0.05] were obviously increased.Moreover,there was a postive correlation between betatrophin and HOMA-IR (r=0.425,P＜0.05) as well as 25(OH)D3 (r=0.577,P＜0.05) and Ca2+ (r=0.448,P＜0.05),while the betatrophin was negatively related to BMI (r=-0.451,P＜0.05),triglycerides (r=-0.454,P＜0.05),LDL-C (r=-0.551,P＜0.05).Circulating betatrophin levels were higher when 25 (OH) D3 levels were beyond 35 nmol/L in PCOS patients [(0.539±0.092) ng/ml vs.(0.199±0.031) ng/ml,t=3.072,P＜0.001],and the multiple linear regression analysis showed that the main factors affecting the levels of betatrophin were BMI (OR=-0.260),HOMA-IR (OR=0.218),25(OH)D3 (OR=0.238),and glycerol (OR=-0.162).Conclusion It is speculated that betatrophin may be a valuable predictive factor of PCOS because betatrophin is closely associated with insulin resistance,lipid metabolism disorders and the lack of 25-hydroxy vitamin D3 in PCOS patients.