Objective To introduce the techniques and specification of complete-block radical gastrectomy, and to compare with traditional radical gastrectomy in the operative time, surgical costs and postoperative complications. Methods Thirty gastric cancer patients meet the criteria were randomly divided into 2 groups. Group A: complete-block radical gastrectomy;group B: radical gastrectomy. Analysis and comparison were made on tumor site, time consuming of block, the proportion of gastrointestinal reconstruction, operative time, cost of block, pathological TNM stage, length of stay, postoperative complications and postoperative chemotherapy. Results Group A has longer operative time than group B (P<0.05). However, there was no significant statistical difference in the other indicators between these 2 groups. Conclusions Complete-block technology can separate cancer more completely during operation with the advantage of simplicity, low-cost, little effect on the subsequent operation. The incidence of postoperative complications and length of stay with the traditional surgery there was no significant difference.

Adenocarcinoma ; metabolism ; pathology ; Humans ; Stomach Neoplasms ; metabolism ; pathology ; alpha-Fetoproteins ; metabolism

Objective To obtain the antigen and antibody of JC virus(JCV)VP2.Methods The JCV VP2 gene were amplified from a cerebrospinal fluid sample by polymerase chain reaction (PCR)and confirmed by sequencing.Then,the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET-32a(+)-VP2.The recombinant plasmid was transformed into the competent E.coli BL21.Induced with isopropyl-β-D-1-1 thiogalactopyranoside (IPTG),E.coli BL21 were subsequently crushed by ultrasound.The gene expression in the supernatant was analyzed by Western blot.Thereafter,the expressed protein was purified by isoeleetric point method.The polyclonal antibody against JCV VP2 protein was obtained from the BALB/c mouse immunized with the purified protein.Results The VP2 fusion protein was expressed in the E.coli BL21.The recombinant fusion protein was expressed by IPTG induetion with relative molecular mass of 58.5×10~3.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDSPAGE)analysis showed that the expression level was highter after 6-10 h of IPTG induction.The recombinant protein had good antigenicity which was confirmed by BALB/c mice immunized with the protein.Conclusions The successful expression and purification of VP2 fusion protein and the antibody will be valuable for the study on the biological function of VP2 and JCV epidemiologieal investigation.

Objective To explore the influence of fluorine on mRNA and protein expression of the insulin-like growth factor-1 (IGF-1) and its receptor of rat osteoblasts.Methods Osteoblasts were isolated from rat bone by enzyme digestion.Different fluorine concentration [0 (control),10-7,10-6,10-5,10-4,10-3 mol/L] was add to the second generation osteoblasts.The IGF-1 in the culture medium was determined by radioimmunoassay (RIA) at different fluorine concentration and different time (24,48 h).The expression of IGF-1 receptor was measured by the method of fluorescent quantitation PCR and the expression of protein IGF-1 receptor was measured by Western blotting.Results ①With increased dose of fluoride exposure,IGF-1 concentration in the osteoblastic culture medium increased first and then decreased at 24,48 h,respectively.Compared to the control group [(38.83 ± 3.48)ng/L],IGF-1 concentration of the 24 h 10-6 mol/L group[(65.45 ± 4.84)ng/L] was higher,and the difference was statistically significant(P ＜ 0.05).The same result was also shown in the 48 h 10-5 mol/L group [(59.14 ± 1.53)ng/L] to its corresponding control group [(33.79 ± 1.84)ng/L,P ＜ 0.05].②The mRNA expression of IGF-1 receptor of the 24,48 h 10-5 mol/L groups (0.0055 ± 0.0004,0.0262 ± 0.0040) was significantly higher than their corresponding control groups (0.0022 ± 0.0001,0.0073 ± 0.0008,all P ＜ 0.05).③With increased dose of fluoride exposure,the protein expression of IGF-1 receptor increased first and then decreased ;the expression of 24 h 10-5 mol/L group (1.39 ± 0.16) was compared with the corresponding control group (0.86 ±0.12),and the difference was statistically significant (P ＜ 0.05) ; the expression of 48 h every fluorine group was also compared with the corresponding control group,and the difference was not statistically significant(all P＞ 0.05).Conclusions Fluorine can affect the mRNA and protein expression of osteoblastic IGF-1 and its receptor.It indicates that IGFS signal transduction pathways play an important role in fluorine regulation of bone metabolism.

ObjectiveTo investigate the expression and clinical value of coiled-coil domain-containing protein 34 (CCDC34) in hepatocellular carcinoma (HCC), and to predict the role of CCDC34 in the development and progression of HCC. MethodsThe datasets of HCC were downloaded from The Cancer Genome Atlas (TCGA) to obtain the expression profile and clinical information of the CCDC34 gene. The bioinformatics method was used to analyze the expression of CCDC34 in HCC, its correlation with clinicopathological parameters, and its influence on prognosis. The gene set enrichment analysis (GSEA) was used to predict the possible pathways regulated by the CCDC34 gene in HCC. The independent samples t-test and the paired t-test were used for comparison of continuous data between two groups; the Kaplan-Meier method and the log-rank test were used for survival analysis; the Cox proportional-hazards regression model analysis was used to investigate the influencing factors for prognosis. P＜0.01 was the standard for judging significant enrichment in GSEA, and the false discovery rate was ＜0.05. ResultsIn TCGA database, CCDC34 was highly expressed in tumor tissue, and there was a significant difference in the expression of CCDC34 between patients with different TNM stages and tumor grades (t=2.118 and 3.622, P=0.035 and P＜0.001). The patients with high expression of CCDC34 had a significantly shorter overall survival time than those with low expression (χ2=21.716, P＜005). The multivariate Cox regression analysis showed that the expression of CCDC34 （HR＝2.287,95%CI:1.312-3.987）and TNM stage(HR=1.943,95%CI:1.101-3.429) were independent risk factors for the overall survival time of patients with HCC (all P＜0.05). The enrichment of 8 pathway gene sets, including base excision repair and spliceosome, was observed in the samples with high expression of CCDC34 (P＜0.01, FDR ＜0.05). ConclusionCCDC34 may play a vital role in the development and progression of HCC and thus become a new prognostic indicator and a potential therapeutic target.

OBJECTIVENitric oxide (NO) is a key factor for penile erection. Its generation is mainly regulated by nitric oxide synthase (NOS), while both phospho-Erkl/2 (P-Erkl/2) and phospho-Aktl (P-Aktl) can affect the expression and activity of NOS and consequently penile erection. This experiment aimed to study the expressions of P-Erkl/2 and P-Aktl in the corpus cavernosum and their possible roles in erectile dysfunction in aged rats.

METHODSWe included 10 two-month-old male SD rats in Group A and another 10 eighteen-month-old ones in Group B, measured the levels of serum testosterone, and detected the expressions of P-Erkl/2 and P-Aktl in the corpus cavernosum by immunohistochemistry and RT-PCR.

RESULTSCompared with Group A, Group B showed a significantly decreased level of serum testosterone (9.57 +/- 1.57 nmol/L vs 4.73 +/- 0.94 nmol/L, P < 0.05), and remarkably increased mRNA expressions of P-Erk1 and P-Erk2 and protein expression of P-Erkl/2 (0.47 +/- 0.09, 0.61 +/- 0.11 and 7.50 +/- 1.81 vs 0.95 +/- 0.06, 0.92 +/- 0.05 and 32.09 +/- 8.45, P < 0.05). But there were no significant differences in the mRNA and protein expressions of P-Akt1 between the two groups (0.97 +/- 0.04 and 11.67 +/- 5.61 vs 0.94 +/- 0.05 and 10.93 +/- 3.06, P > 0.05).

CONCLUSIONThe overexpression of P-Erk1/2 in the corpus cavernous may be one of the important mechanisms of aging-related erectile dysfunction.

Aging ; Animals ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Penis ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Kesuting Capsules (a fast cough suppressant,containing Ephedrae Herba,Papaveris Pericarpium,Platycodonis Radix,etc.).METHODS The analysis of trichloromethane-strong ammonia extract of Kesuting Capsules was performed on a 35 ℃ thermostatic Welch Ultimate(◎) XB-C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.01 mol/L potassium dihydrogen phosphate buffer flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 210 nm.RESULTS Morphine,ephedrine hydrochloride,pseudoephedrine hydrochloride and codeine phosphate showed good linear relationships within the ranges of 8.054-67.12 μg/mL (r =0.999 5),22.31-185.9 μg/mL (r =0.999 9),21.26-177.2 μg/mL (r =0.999 7) and 1.212-10.09 μg/mL (r =0.999 7),whose average recoveries (n =9) were 100.9％ (RSD =2.0％),101.4％ (RSD =3.6％),105.3％ (RSD =1.2％) and 106.2％ (RSD =1.2％),respectively.CONCLUSION This simple method can be used for the rapid quality control of Kesuting Capsules.

AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.

In the areas of prevention and life skills counseling for breast cancer, risk assessment and prediction can assist clinicians to decide if chemoprevention or prophylactic surgery is needed or suggestions on improving the quality of life for their clients. Several mathematical models, namely Gail Model, Claus Model, BRCAPRO Model and Cuzick-Tyrer Model etc. have been developed to make predictions, clinically. This paper has reviewed the development, operation, advantage versus disadvantage and areas of application for the four models. Having family history of breast cancer, one subject was calculated on the risks by the four models and different results were found. Up to 45 years old, the accumulative risks from the four models and population risk were 1.9%, 11.8%, 2.5%, 5.0% and i.6%, respectively. To 75 years old, they were 20.2%,32.5%, 13.1%, 25.0% and 8.5%, respectively. The subject had a relatively high breast cancer risk during her lifetime. A new model is supposed to include a variety of important risk factors and to be validated by large scale of case-control samples. Incidence of breast cancer in China had significantly increased during the last ten years, but the research on developing assessment methods of breast cancer risk had never been reported, suggesting that the development of models for Chinese population is necessary.

Objective To investigate the optimal scanning parameters for diffusion tensor imaging of the cervical cord.Methods MRI and diffusion tensor imaging of the cervical cord was performed in 80 healthy adult volunteers.Different parameters including b values,the number of the diffusion sensitive gradient directions,the number of excitations,and slice thickness were applied and their effects on the quality of the images were compared.DTI was performed on the cervical spinal cord with different b-values (400,700,and 1000 s/mm~2)in group 1,with different numbers of diffusion gradient directions(6,13, and 25)in group 2,with different numbers of excitations(2,4,and 8)in group 3,and with different slice thicknesses(2,3,and 4 mm)in group 4.Two radiology experts gave a score to every image with double blind methods,then compared the image quality.Results In the comparison of the four different parameters,DTIs using a b value of 700 s/mm~2(2.25?0.58)showed better image quality than those with the b values of 400 s/mm~2(1.86?0.53)and 1000 s/mm~2(1.48?0.35)(P