Objective To explore the value of glypican-3(GPC-3)mRNA and paternally expressed 10(PEG10)mRNA in peripheral blood in diagnosis of metastasis in hepatocellular carcinoma(HCC). Methods With SYBR Green I as fluorescence signal,real-time fluorescent quantitative reverse transcription-polymerase chain reaction was employed to detect the expression of GPC-3 mRNA and PEG10 mRNA in peripheral blood from patients with HCC with metastasis(n=8),HCC without metastasis(n=12)and hepatic cirrhosis(n=11),and receiver operator characteristics curve(ROC)and specific parameters were adopted to analyse their value in predictive and exclusive diagnosis. Results The expression of GPC-3 mRNA and PEG10 mRNA in HCC with metastasis was significantly higher than that in HCC without metastasis and in hepatic cirrhosis(P<0.05),while there was no significant difference in the expression of GPC-3 mRNA and PEG10 mRNA between HCC without metastasis and hepatic cirrhosis.In single test,the sensitivities in the differential diagnosis between HCC with metastasis and HCC without metastasis were 66.7%for GPC-3 mRNA and 72.2%for PEG10 mRNA,and the specificities were 91.7%and 91.7%.respectively.The areas under ROC were 0.748 for GPC-3 mRNA and 0.812 for PEG10 mRNA.With two markers in parallel test,the sensitivity,specificity,negative likelihood and diagnostic accuracy were 90.7%,84.O%,0.11 and 83.3%,respectively.In serial test,the sensitivity,specificity,positive likelihood and diagnostic accuracy were 60.5%,98.7%,45.5 and 73.3%,respectively. Conclusion Detection of GPC-3 mRNA and PEG10 mRNA in peripheral blood may help to predict blood metastasis and extrahepatic metastasis of HCC,and PEG10 mRNA works better than GPC-3 mRNA.The serial test of GPC-3 mRNA and PEG10 mRNA is helpful to the predictive diagnosis of peripheral blood metastasis of HCC.

BACKGROUND: Currently, acid etching and bonding technology have been widely used in clinical stomatology. Data have indicated that the main content of inorganic elements (calcium and phosphorus) has a certain difference between the dental enamel and dentin of the young and adult permanent teeth. OBJECTIVE:To measure the content of main inorganic elements, calcium and phosphorus, in the dental enamel and dentin from young and adult permanent teeth with spectrophotometry and ethylene diamine tetraacetie acid titration method. METHODS:Each 20 adult and young permanent teethin vitrowere selected. Plaster stone and water was mixed; when it was nearly dried, the teeth were verticaly cast in the mixture and the tooth surface was exposed. The models were cut into the slices using syj-200 precision cutting machine, and then the slices were put into nitric acid and dissolved through heater to prepare standard solution. At last, the concentrations of calcium and phosphorus in the hard tissue of both young and adult permanent teeth were measured with spectrophotometric method and titration method. RESULTS AND CONCLUSION:The content of calcium and phosphorus and calcium/phosphorus ratio in the hard tissue of young permanent teeth were less than those of adult permanent teeth (P < 0.05), reflecting that the organic matter content was more than that in adult permanent teeth, but their mineralized degree was inferior to that of adult permanent teeth (P < 0.05). These findings indicate that the young permanent teeth are more acid proof than the adult permanent teeth; therefore, the acid etching time can be properly prolonged for young permanent teeth in clinical treatment, in order to achieve better effects.

Objective To explore the possible mechanism of repetitive transcranial magnetic stimulation (rTMS) in learning and memory ability of Alzheimer's disease (AD) rats. Methods 90 SD rats with no differ-ence in learning and memory were selected and 30 rats were randomly selected as control group. The rest 60 were used for establishment of AD model by injecting A beta 1-42 into bilateral hippocampus of rats ,which was verified by Morris water maze test and immunohistochemistry. The successfully prepared model rats were randomly divided into AD group and (rTMS+AD) group,30 rats in each group. The (rTMS+AD) group was treated with rTMS,and the control group and AD group with pseudo stimulation. After intervention,Water maze test,EEG acquisition and analysis,immunohistochemistry and statistical analysis were done. Results (1)Compared with control group, the escape latency of AD group was longer and the average distance from the platform was farther away. Compared with AD group,the escape latency of (rTMS+AD) group was significantly longer and the average distance from the platform was significantly farther away (P<0.05).(2)Compared with control group,the LZ complexity of EEG signals in AD group was decreased. Compared with the AD group,the LZ complexity of EEG signals in (rTMS+AD) group was significantly increased statistically(P< 0.05).(3)The results of immunohistochemistry showed that there was no plaque in the brain tissues of control rats. The morphology of the cells was clear ,and the cells arranged neatly. Myelin staining was deeper and the distribution was long andconnected. There was plaque in the brain tissues of AD rats. Brain tissue cells became atrophy ,the morphology of the cells was not clear ,and the cells arranged randomly. Myelin staining was lighter and the distribution was short and disconnected. Compared with AD group,the cell atrophy of (rTMS+AD) group was reduced,and Myelin staining became deeper. Conclusions rTMS can increase the distribution of myelin in brain tissue ,raise the LZ complexity of EEG signals in AD model rats , and improve the learning and memory ability of them.

Transcatheter arterial chemoembolization (TACE) was the preferred method of non-operation treatment for hepatocellular carcinoma (HCC).Radical resection of HCC remains difficult,extrahepatic metastasis was not easy to deal with,and repeated treatment aggravated the liver injury,so the long-term efficacy was poor.Sorafenib could control tumor angiogenesis and block the proliferation of tumor cells.A male patient with primary HCC and in the stage Ⅱb according to the Chinese clinical liver cancer staging system was treated by TACE combined with sorafenib and antiviral treatment in Henan Cancer Hospital.After the treatment,the intrahepatic lesions were inactive,and the pulmonary metastasis was partially relieved.The patient was followed up till May 2012,and the survival time was 39 months.The hepatic function was normal,and the hepatitis B virus (HBV) replication was negative.No intervention treatmentrelated complications were detected,and the KPS score was 100.

Aim To establish a HPLC method for de-termining cimifugin in rat plasma and investigate the pharmacokinetic characteristics of cimifugin in rats. Methods The plasma concentration of cimifugin was detected by HPLC in acetonitrile protein precipitation method after intragastric administration of cimifugin. The pharmacokinetic parameters were calculated by the procedure of DAS 2 . 1 . Results The regression equa-tion of cimifugin in rats plasma was Y =0. 187 X -0. 0236 (R2 =0. 998 2),which shows a good linear re-lation at 1 - 70 mg · L-1 . The concentration-time curves conformed to two-compartment model. The main pharmacokinetic parameters of Tmax, Cmax, T1/2α, T1/2z, Vd ,AUC(0-t) and AUC(0-∞) were 80 min, 10. 359 mg ·L-1 , 93. 131 min, 2. 179 L · kg-1 , 1946. 085 mg ·L-1 · min, 2138. 57 mg · L-1 · min, respectively. Conclusions We established a HPLC method to de-termine the concentration of cimifugin in plasma. The method is so highly specified and sensitive that it can be used in quantitative analysis in vivo on cimifugin. Cimifugin can be rapidly absorbed, reach the highest concentration and produce effect.

Purpose To explore the antiviral effect of interleukin 29(IL-29) on hepatitis B virus in vitro.Methods To study the antiviral effect of IL-29 against hepatitis B virus by the amount of HBV mRNA detected .Through the quantity of mRNA translated from genes of MxA,2′,5′-OAS,PKR and RNase L as well as the signal pathway induced by IL-29,we used RT-PCR and Western blot to discuss the anti-hepatitis B virus mechanism which was stimulated by IL-29.Results The amount of HBV mRNA in HepG2.2.15 cells was reduced by stimulation of IL-29.The expression of MxA and 2′,5′-OAS was up-regulated,as well as P-ERK and P-AKT were activated by IL-29.Conclusion These findings showed that IL-29 had obvious antiviral activity towards HBV in HepG2.2.15 cells.

Purpose To investigate the effect of interleukin-10(IL-10)on IL-15mRNA and IL-6mRNA in Hela cells induced by lipopolysaccharide(LPS)and to analyze their activiated signal transduction pathways.Methods Extracted total RNA and total proteins of cultured Hela cells,which were treated with different concentrations of LPS,or IL-10 alone or in combined use,were to analyze the levels of IL-15mRNA and IL-6mRNA using RT-PCR and to analyze the expression of signal transduction pathway proteins using Western blot.Results RT-PCR indicated that expression of IL-15mRNA and IL-6mRNA in Hela cells strikingly increased after 12 h using 1 ng-10μg LPS(P＜0.01 vs control),which had dose-dependence and achieved peak value using 100 ng/mL.During 0-24 h,expression of IL-15mRNA and IL-6mRNA strikingly increased with time changing(P＜0.01 vs control),which had time-dependence and achieved peak value at 12 h.Expression of IL-15mRNA and IL-6mRNA had no conspicuous difference in Hela cells treated with IL-10(10 ng/mL)alone(P＞0.05 vs control).Different concentrations of IL-10 1,10,100 ns/mL)markedly downregnlated expression of IL-15mRNA and IL-6mRNA in Hela cells induced by LPS.Furthermore,the effect of inhibition will be more obvious with dose increasing.Western blot indicated that LPS upregulated expression of IL-15mRNA and IL-6mRNA by phosphorylation of PI3K/AKT and ERK1/2.IL-10 blocked phosphorylation of AKT,but could not affect phosphorylation of ERK1/2.Conclusion IL-10 downregulated the expression of inflammatory cytokines IL-15 and 1L-6 induced by LPS,which may correlate with the fact that phosphorylation of AKT was blcoked by IL-10.Therefore,IL-10 may be used to prevent and treat some clincal infective diseases.

Aim To explore the expressed level of ini-tiative key factors TSLP and IL-33 in a human kerati-nocyte cell line,HaCaT cells were chosen to be stimu-lated by different stimulants,and develop a stable and effective in vitro model to observe allergic sensitization. Methods HaCaT cells were cultured in K-SFM with different stimulants to screen out the stimulants which could significantly improve the expressed level of TSLP and IL-33.Expressed level of TSLP and IL-33 was an-alyzed by ELISA kits and immunofluorescence.Re-sults (1 )The dose-response relationship of single stimulant indicated that both Poly(I:C)and TNF-αcould significantly improve expressed level of TSLP and IL-33 in HaCaT cells,but the rest of stimulants was not observed significant stimulation in concentration range of this experiment.(2)Dose-effect relationship of combined stimulants indicated that poly(I ∶C)1 00 mg·L -1 combined with TNF-α20 μg·L -1 was the most efficient.(3)Time-effect relationship of the a-bove-mentioned combined stimulants showed that 1 2 h was the optimal time of stimulation.Conclusions Different stimulants and different time result in various expressed levels of TSLP and IL-33 in HaCaT cells.1 2 h stimulus duration of Poly(I:C)1 00 mg·L -1 com-bined with TNF-α20 μg · L -1 is the most efficient stimulating way.This result provides an effective in vitro model to study the pathomechanism and drug effi-cacy of allergic sensitization.

Objective To discuss the function of pelvic lymphadenectomy in radical cystectomy. Methods Ninety-five patients with bladder cancer (76 males and 19 females) underwent radical cys-tectomy. Clinical data were reviewed. Median age was 62 years old (25-78). Among all patients, 49 were newly diagnosed and 46 had recurrent disease. Of 95 patients, 87 were urothelial cell carcinoma, 5 were adenocarcinoma, and 3 were squamous cell carcinoma. Of 87 urothelial cell carcinoma cases, 17 were grade 1, 39 were grade 2, and 31 were grade 3. Of 95 patients, 10 were Ta-T1,54 were T2 ,26 were T3 ,and 5 were T4 according to AJCC classification. All cases accepted bilateral pelvic lymphade-nectomy according to standard protocol. Results Bilateral lymphadenectomy was taken an average time about 20 min. No important vessels and nerves injury occurred and average bleeding volume was 25 ml during procedure. A median of 10 lymph nodes were removed (range, 1-20). The nodal posi-tive rate was 17.9% (17/95) with 58.8% (10/17) bilateral lymph nodes positive. Short-term opera-tion-related complication rate was 12.6% (12/95). No operation-related death happened. Median fol-low up time was 34 months (3 to 64 months). Sixteen cases died during followup and the 3-year over-all survival rate was 84.5%. Conclusions Bilateral pelvic lymphadenectomy should be routinely per-formed during radical cystectomy. Standard lymphadenectomy could document accurately the staging and improve the overall survival in radical cystectomy without severe complications.

AIM:To explore the inhibitory effects of tumor associated mitochondrial protein 12 (TAMP12) on tumor cell apoptosis. METHODS: (1) A retrovirus expression vector was recombinated and transfected into the packaging cell line PA317. The virus particles were obtained to infect the target cell line HepG2 low expressing of TAMP12. The expression of TAMP12 mRNA was detected by RT-PCR. The subcellular localization and quantification of TAMP12 protein labeled with double fluorescein were observed under confocal laser scanning microscope (CLSM). (2) Hoechst33258 staining and flow cytometry (FACS) were used to analysis the apoptosis of HepG2 cells treated with 5-fluorouracil (5-FU). RESULTS: (1) The CLSM observation showed that TAMP12 protein was mainly expressed in mitochondria of HepG2 cells. The expressions of TAMP12 gene and protein were stable and high in transfected HepG2 cells. (2) Upon treatment with 5-FU, the transfected HepG2 cells showed a fairly integrated nucelus while the control HepG2 cells exhibited chromatin condensation, marginalization and karyorhexix. Moreover, the apoptosis rate of transfeced HepG2 cells was significantly lower than that in control HepG2 cells (P