AIM: To illuminate the therapeutic basis of Compound Wurenchun Capsules in combination with serum pharmacology,the lignans of this preparation migrating to blood of rats after oral administration having been accomplished. METHODS: By UPLC-MS/MS method, lignans migrating to blood were affirmed by comparing the extracted ion chromatography (EIC) of the serum containing drug with the ones of Compound Wurenchun Capsules, the control serum and control articles, correlated ion peak in mass-spectrogram were analyzed at the same time. RESULTS: Five lignans migrating to blood have been found, they are schisandrin, schisandrol B, schisantherin, deoxyschizandrin and schisandrin B. CONCLUSION: Five lignans above mentioned are likely the effective substances of Compound Wurenchun Capsules in human body. More study by means of combining with serum pharmacology will illuminate the therapeutic basis of this preparation.

0.05),but the efficacy index was higher in groups B and C than that in group A(P

Objective To identify the drug-induced constituents in rat serum containing drug of Compound Wurenchun Capsula and determine the content of these constituents. Methods Identification of the drug-induced constituents in serum has been carried out by combinative method of HPLC-DAD and UPLC-MS/MS. The content of four lignans in serum has been detected by HPLC-UV. Results Seven of eight original form compounds in serum have been identified as schisandrin,gomisin J,schisandrol B,deoxyschizandrin,gomisin N,schisandrin B,and schisandrin C. The UV spectrogram of five metabolites showed the absorption character of dibenzocyclooctadiene lignans. Eight lignans were identified by UPLC-MS/MS,besides schisantherin,there are seven lignan-like ones detected by HPLC-DAD. The content of schisandrin,schisandrol B,deoxyschizandrin,and schisandrin B in serum was (8.145 3?1.020 2),(6.604 5?1.341 4),(0.560 1?0.137 5),and (5.933 0?0.966 6) ?g/mL,respectively. ConclusionLignans and their metabolites are composed of the main drug-induced constituents in rat serum.

Objective To investigate human cell division control protein 4(hCDC4) expression and its correlation with the clini‐copathological features in oral squamous cell carcinoma(OSCC) .Methods We freshly collected 52 samples of surgically resected OSCC tissues and 12 samples of normal tissues .hCDC4 expression in the samples was detected by immunohistochemical staining . The correlation between hCDC4 protein expression and clinicopathological feature was analysed .OSCC cells and Tca8113 were transfected with hCDC4‐siRNA ,cell proliferation and c‐Myc and Cyclin E protein expression were determined by using M TT and Western blot .Results The hCDC4 protein expression in normal tissues was significantly up‐regulated compared to those in OSCC tissues (83 .3% vs .25 .0% ,P < 0 .05) .Clinicopathological analysis revealed that reduced hCDC4 expression was associated with large tumor size ( ≥ 4 cm) and high clinic stage ( Ⅲ + Ⅳ ) (P< 0 .05) .hCDC4 knockdown by siRNA led to increased cell prolifera‐tion and c‐Myc and Cyclin E protein accumulation in Tca8113 cells .Conclusion Loss of hCDC4 may promote tumor progression by resulting in c‐Myc and Cyclin E protein accumulation in OSCC .

Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.

OBJECTIVE To evaluate the value of central compartment neck dissection for cN0 papillary thyroid carcinoma.METHODS The clinical data of 46 cases with cN0 papillary thyroid carcinoma in our hospital from Jan.1999 to June 2004 were retrospectively studied.The level Ⅵ lymph nodes were removed during the operation.RESULTS In 46 cN0 cases,there were 11 cases(23.9 %) pathologically proved lymph nodes metastasis after operation.Four patients developed neck recurrence and underwent neck dissection.CONCLUSION It is necessary to probe the level Ⅵ lymph nodes during the operation for cN0 papillary thyroid carcinoma patients.

Objective To investigate the modulation of EPCs by interleukin 1β (IL-1β) and p38 mitogen activated protein kinase (p38MAPK) and the pathogenesis resulting from their dysdifferenfiation after trauma.Method Thirty pigs were divided into a control group (n = 15) and a multiple organ dysfimction syndrome (MODS) group (n = 15), the latter of which were subjected to a "two-hit" injury including hemon'hagic shock and endotoxemia. Phosphorylation of p38MAPK in peripheral blood mononuclear cells was monitored by western blotting. The concentration of IL-1β in peripheral blood plasma was determined by ELISA and the numbers of EPCs with FCM in peripheral blood plasma were monitored. The morbidity rates in the two groups were compared by chi square test. The levels of phosphorylation of p38MAPK in peripheral blood mononuclear cells, the concentmtions of IL-1β in peripheral blood plasma and the numbers of EPCs in the peripheral blood were compared between groups using with Student's t lest. Results The level of p38MAPK phosphorylation was more augmented and the concen-tration of IL-1β higher in peripheral blood mononuelear cells and plasma from MODS pigs compared with those from control pigs; nevertheless the mauler of EPC conspicuously decreased in the peripheral blood (P <0.01). The morbidity rate in the MODS group was much higher than that in the control group (P < 0.01). There were fewer EPCs in the peripheral blood of animals in group M than in the peripheral blood of animals in group C (P <0.01). Conclusions p38MAPK phosphorylation is important for the pathogenesis of MODS. p38MAPK phospho-rylation might cause the concentration of IL-1β in the peripheral blood plasma to rise and could cause a drop in the numbers of EPCs, thereby aggravating the inflanmmatory reaction in MODS.

Abstrate:Aim To construct HEK293 cell line with stable and high expression of sodium taurocholate cotransporting polypeptide (NTCP ) efficiently and rapidly.Method Vector expressing EGFP-NTCP fusion protein was constructed and verified by DNA sequencing.The pEGFP-NTCP expression vector was transfected into HEK293 cells by FuGENE 6 transfection reagent. The transfected cells with high expression of green fluorescent protein were selected using fluorescence microscope for screening of G418 for 14 days to obtain cell lines stably and highly expressing NTCP.NTCP expression was detected by RT-PCR,qRT-PCR, Western-blot and the uptake experiment of taurocholic acid.Re-sult RT-PCR,qRT-PCR,Western-blot and the uptake experi-ment revealed that compared to the control cells,the expression of NTCP was significantly positive (P<0.01)in stable trans-fected cells showing green fluorescence (P<0.05 ).Conclusion The HEK293 cell line with stable and high expression of NTCP has been established efficiently and rapidly,which provides a cellular model for the study of the mechanism of the uptake of bile acid derivatives.

Objective To compare the safety and efficacy of direct and remedial rotational atherectomy in the treatment of heavily calcified coronary artery lesions.Methods We retrospectively reviewed 58 patients admitted in the Shanghai Chest Hospital and Liaocheng People Hospital from May 2012 to July 2015 who had received stent implantation and rotational atherectomy.The 58 patients were divided into two groups which were the direct atherectomy group (n =27) and the remedial atherectomy group (n =31).General clinical date,lesion and procedural characteristics,intraoperative complications,in-hospital and follow-up MACCE were compared between the two groups.Results There were no differences between the two groups in general clinical date intraoperative complications,amount of contrast agent used,proceduraltime,rates of in-hospital and follow-up MACCE.Nevertheless,compared with the direct artherectomy group,the remedial group had more number of balloon dilations during procedure [3 (1,5) vs.2 (1,2),P ＜ 0.001] and higher peak cardiac troponin levels [1.1 (0.3,3.0) μg/L vs.0.5 (0.1,2.3) μg/L,P =0.032].Conclusions Remedial rotational atherectomy with drug-eluting stent had the same safety and efficacy as direct atheretomy with drug-eluting stent in treating patients with heavily calcified coronary lesions.It is reasonable and safe to transform routine PCI to remedial rotational atherectomy when the 2.0 mm semi compliant balloon or/and 2.5 mm non-compliant balloon cannot pass through or dilate the lesions.

Objective:To investigate the effect of long non-coding (lnc) RNA HCP5 on the radiation sensitivity of glioma cells and underlying mechanism.Methods:The glioma cells U251 and U87 were irradiated with 0, 2, 4, 6, and 8 Gy rays as different doses.si-Con, si-HCP5, pcDNA, and pcDNA-HCP5 were transfected into cells U251 and U87, recorded as si-con group, si-HCP5 group, pcDNA group, and pcDNA-HCP5 group.si-Con and si-HCP5 were transfected into cells U251 and U87, and then irradiated with 4 Gy rays, respectively, recorded as IR+ si-con group and IR+ si-HCP5 group, the cells only irradiated with 4 Gy rays were recorded as IR group.After si-HCP5 with anti-miR-con and anti-miR-508-3p was co-transfected into cell U251 and U87, respectively, irradiated with 4 Gy rays, recorded as IR+ si-HCP5+ anti-miR-con group and IR+ si-HCP5+ anti-miR-508-3p group, respectively, the transfection was performed by liposome method.RT-qPCR was used to detect the expression of miR-508-3p and HCP5.Cell clone formation assay was used to detect the radiosensitivity of glioma cells.Flow cytometry was used to detect apoptosis, dual luciferase Reporter gene detection experiments detects fluorescence activity.Results:HCP5 was highly expressed in radiation-treated glioma cells, and miR-508-3p was lowly expressed.After silenced HCP5, U251 and U87 cells had enhanced radiosensitivity and apoptotic rate((16.67±1.68) vs (3.58±0.62), t=21.929, P<0.05; (12.32±1.08) vs (4.48±0.71), t=18.198, P<0.05) was increased, and γ-H2AX( (0.45±0.04) vs (0.23±0.05), t=10.307, P<0.05; (0.38±0.04) vs (0.24±0.03), t=8.400, P<0.05), Cleaved caspase-3((0.37±0.04) vs (0.16±0.03), t=12.600, P<0.05; (0.38±0.04) vs (0.22±0.03), t=9.600, P<0.05) expressions were increased.Compared with silencing HCP5 or radiation treatment alone, silencing HCP5 and radiation treatment of U251 cells simultaneously, the apoptosis rate ((25.34±1.54) vs (16.67±1.68), t=11.413, P<0.05; (25.34±1.54) vs (11.13±1.06), t=22.802, P<0.05) was significantly increased, and γ-H2AX((0.69±0.05) vs (0.45±0.04), t=11.245, P<0.05; (0.69±0.05) vs (0.31±0.04), t=17.804, P<0.05), Cleaved caspase-3 ((0.52±0.06/0.37±0.04, t=6.240, P<0.05) (0.52±0.06/0.34±0.04, t=7.488, P<0.05) expressions were increased.The expressions of p-PI3K ((0.21±0.02) vs (0.52±0.04), t=20.795, P<0.05; (0.26±0.23 ), ( 0.67±0.07), t=5.116, P<0.05), p- AKT ((0.22±0.03) vs (0.66±0.07), t=17.332, P<0.05; (0.23±0.04) vs (0.71±0.03), t=28.800, P<0.05) in U251 and U87 cells were decreased.HCP5 can target the regulation of miR-508-3p expression; interfering with miR-508-3p reversed the effects of silent HCP5 and radiation on the radiation sensitization and apoptosis of U251 and U87 cells.It reduced the expression levels of reducing γ-H2AX and Cleaved caspase-3, while increased the expression levels of p-PI3K and p-AKT. Conclusion:Silencing lncRNA HCP5 can enhance the radiation sensitivity of glioma cells and promote apoptosis.The mechanism may be related with the miR-508-3p and PI3K/Akt signaling pathway, which will provide new targets and new ideas for glioma treatment.