Objective To establish a method to determine six kinds of PAEs in human serum simultaneously,and on this basis to understand the level of PAEs in human body.Methods Ultrasonic extraction,gas chromatography-mass spectrometry-selected ion monitoring(GC-MS-SIM) detection and quantitative analysis based on internal standard were used to detect six kinds of PAEs in human serum simultaneously.Finally this method was used in fifty human serum samples.Results The results showed that this method had a good linear relation in the range from 10 to 1 000 ng/ml,and the correlation coefficients of standard curve were higher than 0.999.The average recovery rate and the relative standard deviation(RSD,n=6) of the target compounds in standard-addition blank was 94.5% and 4.4% respectively,the lower limits of detection for DEP,DMP,DBP,BBP,DEHP,DNOP were 0.31,0.85,0.90,0.63,0.88,0.41 ng/ml respectively,and the recovery rates of all samples ranged from 67.98% to 110.8%.Conclusion This method has good recovery rate,reproducibility,lower limit of detection and can be used for the determination of many kinds of PAEs substances in human serum.

Objective To evaluate the efficacy and adverse effects of low doses gemcitabine chemotherapy combined with synchronous high-low oxygen radiotherapy in patients with locally advanced pancreatic cancer.Methods Fifty-six patients with locally advanced pancreatic cancer were randomly divided into two groups by envelop method:radio-chemotherapy group or chemotherapy group.Patients in radio-chmotherapy group were treated with low doses of gemcitabine chemotherapy ( 600 mg/m2 ) combined with high-low oxygen radiotherapy synchronously,paients in chmotherapy group were treated with full doses of gemcitabine chemotherapy ( 1000 mg/m2).The short-term effect,distant metastasis rate,clinical benefit rate,survival rate and adverse events of two groups were observed.Results There was one patient achievedcomplete relief and 15 patients achieved partial relief in radio-chemotherapy group with an overall effective rate of 66.7％ (16/24) ; there were 9 patients achieved partial relief in chemotherapy group with an overall effective rate of 36.0％ (9/25),the difference between the two groups was statistically significant (X2 =4.6082,P =0.0318 ).The clinical benefit rates were 83.3 ％ ( 20/24 ) and 60％ ( 15/25 ),respectively,the difference between the two groups was not statistically significant ( P =0.070).The distant metastasis rates were 66.7％(16/24) and 72％ (18/25),respectively,the difference between the two groups was not statistically significant (P =0.6855).The 12,24 months survival rates were 62.5％ vs 32％,37.5％ vs 12％,the difference between the two groups was statistically significant ( P =0.0325,0.0380).The incidence of serious adverse events was 45.8％ and 4 0 ％ without statistical difference.Conclusions Low doses of gemcitabine chemotherapy combined with high-low oxygen radiotherapy synchronously is better than full doses of gemcitabine chemotherapy with regard to total effective rates and 12,24 months survival rates,with no obvious increase in the incidence of serious adverse events.

Objective:To screen the best extraction process of four medicinal materials in Simiao Junyi ointment by analgesic ex-periments. Methods:The analgesic experiments were performed by the hot-plate test and writhing test in mice to select the best extrac-tion process of four medicinal materials in Simiao Junyi ointment. Results:According to the results of level consistency check, the data of hot-plate test in mice in different groups had no significant difference, and the pesticide effect of Simiao Junyi ointment prepared by different extraction process was similar. However, the data of writhing test had significant difference. The extraction process of the oint-ment with the best analgesic activity was as follows:pepper and rhizoma corydalis were extracted by SFE-CO2 , myrrha and pseudo-gin-seng was extracted by 95% and 50% ethanol, respectively. Conclusion: The optimal extraction process tested by analgesic experi-ments is scientific, reasonable and feasible, and suitable for the research and development of modern traditional Chinese medicine preparations.

Objective To explore influence of thoracic duct ligation on lipid metabolism after esophagectomy. Methods Seventy-four patients with esophageal cancer received esophagectomy were divided into two groups by their thoracic duct ligation, 39 in ligation group and 35 in non-ligation group. All the patients were fed with nutrients through nasal-duodenal tube placed during operation from the 1 st day to the 8th day after surgery, and started taking liquid diet on the 6th day. Blood specimens were collected from the patients on the 1st day after admission, the 9th day, one month and three months after surgery,respectively for biochemical analysis,including determinations of total cholesterol(TC),triglyeeride(TG),high-density lipoprotein-cholesterol ( HDL-C) and low-density lipoprotein-cholesterol ( LDL-C) . Databetween the two groups were compared with statistical analysis. Results No significant difference in plasma levels of TC, TG and HDI.-C was found between the two groups at varied time points. Plasma level of LDL-C in the ligation group significantly decreased on the 9th day after surgery, as compared with that in the nonligation group ( P ＜ 0. 05), and went down to the lowest level one month after surgery. Conclusions Chylomicron was blocked to enter blood stream and production of LDL-C decreased by thoracic duct ligation,which affect lipid absorption and metabolism leading to poor early nutrition in patients with esophagealsurgery due to slower establishment of collateral circulation.

Objective·To investigate the clinicopathologic characteristics and prognosis of adult rhabdomyosarcoma (RMS).Methods·The clinical and pathological data,immunohistochemical test results of 9 RMS were analyzed retrospectively.Results·The information of 6 males and 3 females were collected in this research,whose average age was (46.89+20.09) years old.The tumors had a wide range distribution.The subtypes of histology included 6 cases of embryonal RMS,1 case of alveolar RMS,1 case of spindle cell RMS and 1 case of pleomorphic RMS.Immunohistochemically study showed that 6 cases were myogenin positive,4 cases were MyoDl positive and 8 cases were desmin positive.During the follow-up period of the 7 cases,only 2 patients survived free of tumors,2 patients presented with tumor recurrence and metastasis after the operations,and 3 patients died of tumor.Conclusion·In this study,embryonaI RMS is the most common type,and immunohistochemistry test is helpful to diagnosis of RMS.

OBJECTIVE To investigate the effect ofγ-secretase inhibitor N-[N-(3,5-difluorophen?acetyl)-L-alanyl]-S-phenylglycine t-butyl ester(DAPT)on phenotypic transformation and matrix accu?mulation induced by aristolochic acid(AA) in renal tubular epithelial cells(NRK-52E)and explore the mechanism. METHODS NRK-52E cells were divided stochastically into normal cell control group,AA 10 mg·L-1 group and AA 10 mg·L-1+DAPT 1 and 10μmol·L-1 group. After 24 h,the mRNA expressions of Notch1,Jagged1,Numb,E-cadherin,transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA),bone morphogenic protein 7 (Bmp7),typeⅠ a1 (Col1a1) and Ⅲ collagens a1 (Col3a1)were quantified by quantitative real-time RT-PCR. The protein expressions of Notch1,Jagged1,α-SMA,and Col3a1 in NRK-52E cel s were detected by immunofluorescence staining. RESULTS In NRK-52E cells,AA enhanced the expression of TGF-β1,α-SMA and Col3a1 mRNA(P<0.05),reduced the expression of E-cadherin mRNA(P<0.05),up-regulated the mRNA expression of Notch1 mRNA(P<0.01)and Jagged1(P<0.05),and down-regulated the mRNA expression of Numb mRNA(P<0.05) compared with normal cell control group,indicating that phenotypic transformation and matrix accumu?lation occurred in AA-treated NRK-52E cells,accompanied by activated Notch signaling. Treatment with DAPT inhibited Notch signaling by decreasing the expression of Notch1 and Jagged1 (P<0.05),and increasing the expression of Numb mRNA(P<0.05). Furthermore, DAPT also down-regulated the expression levels of TGF-β1,α-SMA,Col1a1 and Col3a1 mRNA(P<0.05), and up-regulated the expression level of Bmp7 and E-cadherin mRNA(P<0.05) compared with AA group,suggesting that DAPT inhibited phenotypic transformation and matrix accumulation in AA-treated NRK-52E cells. CONCLUSION AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells,which is inhibited by DAPT treatment. The possible mechanism is that DAPT suppresses the activation of Notch signaling,resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition.

Objective:To establish an analytical method for HPLC fingerprint chromatography of Simiao Junyi ointment to provide basis for the quality control standard. Methods:The separation conditions were established to obtain the HPLC fingerprint chromatog-raphy of the main ingredients in Simiao Junyi ointment. The conditions were as follows:the chromatographic column was Ultimate C18-ODS(250 mm ×4.6 mm,5 μm), the mobile phase was acetonitrile-0.1% phosphate solution, the flow rate was 1.0 ml·min-1, the detection wavelength was 280 nm, and the column temperature was 35℃. The common peaks in the chromatography were analyzed for their belongings. Results:Gradient elution was performed under the above optimal separation conditions, the constituents in Simiao Ju-nyi ointment were separated from each other perfectly, and the optimal fingerprint chromatography was obtained. Though the methodolo-gy examination, the indicators such as precision, stability and repeatability of the method were all promising, and the fingerprint chro-matography could be seen clearly and was easy to be analyzed. The relationships between Simiao Junyi ointment and the common peaks of four medicinal materials in the fingerprint chromatography were preliminary determined, which provided important basis for the quali-ty control of Simiao Junyi ointment. Conclusion:The HPLC fingerprint chromatography of Simiao Junyi ointment can be used as an a-nalysis method for the quality control of Simiao Junyi ointment, which provides reference for the quality control standard for the finished product.

Objective:To explore the degradation effect of gamma-rays radiation sterilization on the effective constituents in Simiao Junyi ointment and study the changes of fingerprint chromatography before and after the sterilization to provide basis for the feasibility of gamma-rays radiation sterilization for Simiao Junyi ointment. Methods:The contents of tetrahydropalmatine and PNS ( Panax Notogin-seng saponins) in Simiao Junyi ointment were determined by HPLC, and the fingerprint chromatography was established. The content changes of tetrahydropalmatine and PNS in Simiao Junyi ointment before and after the gamma-rays radiation sterilization were compared among the same batch and various batches, and the relative retention time and relative peak area in the fingerprint chromatography were also compared. Results:The content of tetrahydropalmatine had no change basically before and after the gamma-rays radiation steriliza-tion(P >0. 05), and there was no change in the total content of PNS (P >0. 05). Comparing the HPLC fingerprint chromatography at 280 nm, the relative retention time had statistically significant change after the gamma-rays radiation sterilization ((P <0.05), while the relative peak area had no statistically significant change(P >0. 05). The number of characteristic peaks reduced by one, namely the C8 characteristic peak disappeared in the pasteurized chromatography, and the areas of C6, C9 and C14 peak decreased significantly, while that of C12 increased. Conclusion:Gamma-rays radiation sterilization have no notable effect on the content of tet-rahydropalmatine and PNS in Simiao Junyi ointment, it can be used for the sterilization of Simiao Junyi ointment.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.

OBJECTIVE To investigate the molecular mechanisms of resveratrol( Res)in renal interstitial fibrosis(RlF)in rats with unilateral ureteral obstruction(UUO). METHODS Forty-eight Spra-gur-Dawley rats were randomly divided into UUO( normal saline,n = 16),UUO with Res treatment (Res,20 mg·kg-1 ,n=16),and sham-operation(sham,n=16)models. The kidneys were excised on the 7th and 14th day. The deposition of collagen fiber in the kidney was detected with HE and Masson staining. The levels of sonic hedgehog(SHH,an inducer of SHH pathway)in kidney tissues were deter-mined by ELlSA. lmmunohistochemical analysis was performed to evaluate the protein expression of SHH signaling-related molecules,including SHH,smoothened(Smo),patched-1(Ptch1),and Gli1, proliferating cell nuclear antigen(PCNA)and matrix component typeⅢ collagen. The mRNA expression levels of Smo,Ptch1 and Gli1 were detected by real-time RT-PCR. RESULTS The degree of RlF observed with HE and Masson staining was obviously increased in UUO kidneys,but decreased in Res-treated kidneys. Enhanced expression levels of typeⅢ collagen and PCNA in UUO rats were suppressed by Res treatment(P﹤0.05). Res administration decreased the expression levels of SHH,Smo,and Gli1 (P﹤0.05),but increased the expression of Ptch1(P﹤0.05),suggesting that Res inhibit the obstruction-induced activation of SHH signaling. CONCLUSION Res can attenuate RlF in UUO rats,and the possi-ble mechanism is that Res down-regulates the activity of SHH signaling and inhibits cellular proliferation, resulting in inhibition of matrix accumulation in renal interstitium of UUO rats.