Objective： To develop a RP-HPLC method for the determination of quercetin in UGT1A3 cDNA-transfected cells.
Methods： The lysate of cells transfected with human recombinant uridine 5-diphosphate glucuronosyltransferases UGT1A3 cDNA was co-incubated with quercetin，the reaction was terminated with acetonitrile，and luteolin was used as internal standard. The determination was performed on a C18 reversed phase column with a mobile phase of methanol-0.1% formic acid (V/V) at a flow rate of 1.0 ml/min. The gradient elution was as follows: 0-25 min(30∶70-80∶20,methanol:0.1% formic acid），>25-25.5 min(80∶20)，>25.5-27 min(80∶20-30∶70)，>27-30 min(30∶70). A UV-VIS detector was operated at 368 nm.
Results： The standard curve was linear over the concentration range of 5-200 μmol/L (r=0.9999). The limit of detection was 1.25 μmol/L（S/N≥3），and the limit of quantification was 5 μmol/L (S/N >10，RSD=6.99%). The method afforded recoveries of 99.1%-103.5%，and precisions for inter- and intra-assay were ＜2.5% and ＜8%，respectively. In addition，kinetic analysis indicated that the Km，Vmax and CL int (Vmax /Km) values for quercetin glucuronide were （62.95±13.16）μmol/L，（284.50±24.35）nmol·min-1·g-1 and 4.52 ml·min-1·g-1，respectively.
Conclusion： The method established is accurate and simple and suitable for the determination of quercetin in UGT1A3 cDNA-expressed cells.

Objective： To establish a chiral separation method for determination of fluvastatin enantiomer with in vitro cellular model.
Methods： The determination was performed on Chiralpak AD column（4.6 mm×250 mm）; and the phase consisted of hexane-isopropanol -trifluoroacetic acid (90∶10∶0.1) at a flow rate of 0.5 ml/min with UV detection of 239 nm.
Results： The standard curve was linear over the concentration range of 20 μmol/L-300 μmol/L (r2=0.9993，r2=0.9997). The recovery for this assay was （99.4±0.8 ）%，precision for inter-assay and intra-assay was < 10%.
Conclusion： The normal-phase HPLC chiral separation method was accurate and suitable for study on the stereoselectivity of fluvastatin with in vitro cellular model.

Objective： To establish a comprehensive quality control method for total flavonoid of Fructus Aurantii.
Methods： RP-HPLC and spectrophotometry were applied for the quantitative and fingerprint analysis of total flavonoid of Fructus Aurantii. The contents of naringin and neohesperidin were determined on an Agilent SB-C18 column (4.6 mm×250 mm，5 μm). The mobile phase was composed of 0.02% H3PO4 and CH3CN（80∶20）. The flow rate was 1 ml/min with DAD detected at 280 nm. The column temperature was maintained at 35℃. The fingerprints were developed on an Agilent SB-C18 column(4.6 mm×250 mm，5 μm). The mobile phase was composed of 0.5% HAc and CH3OH with a linear gradient elution. The ratio of 0.5% HAc and CH3OH was: 0 min，80∶20; 10 min，60∶40; 35 min，30∶70; 50 min，0∶100. The flow rate was 1 ml/min with DAD detected at 320 nm. The column temperature was maintained at 30℃. Meanwhile，the contents of total flavonoid were determined at 283 nm.
Results： The contents range of naringin，neohesperidin and total flavonoid were 38.3%-47.2%，21.0%-28.5% and 79.9%-88.6%，respectively. The fingerprints of the effective fractions showed 12 common peaks and the fingerprint similarity was all above 98.0% compared with the standard chromatogram.
Conclusion： The method reported in this paper can be used effectively for the quality control of total flavonoid of Fructus Aurantii.

Objective： To establish a HPLC method for simultaneous determination of 4 effective components from total flavonoids of Scutellaria barbata (FSB).
Methods： The HPLC method was developed on an Agilent Zorbax C18 column (4.6 mm×250 mm，5 μm). The mobile phase was composed of 1% HAc and CH3OH∶CH3CN (80∶20) with a linear gradient elution. The flow rate was 1.0 ml/min，and UV detection wave length was set at 280 nm. The column temperature was maintained at 30℃.
Results： The linear range of 4 effective components (scutellarin,isoscutellarein-8-O-glucuronide，isoscutellerin and luteolin) was 0.14-11.20 μg，0.03-2.40 μg，0.007-0.560 μg and 0.027-2.160 μg，respectively. The average recovery for 4 effective components was（101.9±1.4）%,（103.5±0.6）%,（98.1±2.9）% and（100.5±2.3）%，respectively. The contents of 4 flavonoids were determined，with scutellarin 7.3%-14.3%，isoscutellarein-8-O-glucuronide 2.4%-9.3%，isoscutellerin 0.3%-0.5%，and luteolin 0.2%%-0.6%，respectively.
Conclusion： The method can be used effectively to evaluate the quality of FSB.

Objective： To investigate the optimal alcohol precipitation parameters for extract of Carthamus tinctorius.
Methods： The effects of different factors on the transfer rate of hydroxy safflower yellow A (HSYA) was studied via single factor experiments，including the final alcohol concentration of the liquor，the speed of stirring，the initial density of the extract，the temperature and the pH of the liquor. Based on the results of single factor experiments，the final alcohol concentration of the liquor，the speed of stirring，the initial density of the extract and the pH of the liquor were studied by an orthogonal test and a multiple guidelines grading method，and the transfer rate of HSYA，the yield and the purity of extract in the supernatant were used as comprehensive evaluation index.
Results： The optimal alcohol precipitation process of Carthamus tinctorius extract was as follows: the final alcohol concentration of the liquor 50%，the speed of stirring 500 r/min，the initial density of the extract 1.15 g/ml and the pH of the liquor 5.0.
Conclusion： The proposed alcohol precipitation process is convenient and steady with high transfer rate of HSYA，high yield and purity of extract in the supernatant.

Objective： To investigate the inhibitive and inductive effect of （-）-tetrahydropalmatine (THP) and (+)-THP on main CYP450 isoforms in mouse liver microsomes.
Methods： The in vitro inhibitory effect was evaluated by incubating （-）-THP or (+)-THP with the probe substrates of main phase I metabolic enzymes in mouse liver microsomes，and the remaining substrates were determined by HPLC or LC-MS/MS method. Mice were administered with （-）-THP or (+)-THP at dosage of 240 mg/kg or 60 mg/kg by gastric lavage for successive 7 days，then the cocktail-LC-MS method was applied to assess the activities of main CYP450 isoforms in mouse liver microsomes.
Results： The IC50 values of both （-）-THP and (+)-THP on isoforms studied were higher than 100 μmol/L except that IC50 value of (+)-THP on CYP2C was 43.89 μmol/L，indicating weak inhibition of （-）-THP and (+)-THP on CYP1A2，CYP2D22，CYP2E1 and CYP3A11 in vitro. Compared with the vehicle group，the activities of CYP2D22，CYP2E1 and CYP3A11 were not increased significantly in （-）-THP and (+)-THP treatment groups, while the activities of CYP1A2 in 60 mg/kg and 240 mg/kg （-）-THP groups were 68.7% and 73.0% higher，than that of the vehicle group (P<0.05，P<0.01，respectively)，the activity of CYP2C37 in 240 mg/kg （-）-THP treatment group was 80.4%，higher than that of the vehicle group (P<0.05).
Conclusions： There is negligible or weak inhibition on main CYP450 in mouse liver microsomes by （-）-THP and (+)-THP in vitro. (+)-THP does not induce main CYP450 in mouse liver microsomes while （-）-THP weakly induces CYP1A2 and CYP2C37.

Objective： To develop a gas chromatography method for determination of residual solvents in 7-amino-3-chloro cephalosporanic acid (7-ACCA).
Methods： The residual levels of acetone，methanol，dichloromethane，ethyl acetate，isobutanol，pyridine and toluene in 7-ACCA were measured by gas chromatography using Agilent INNOWAX capillary column（30 m×0.32 mm,0.5 μm). The initial column temperature was 70℃ maintained for 6 min and then raised (10℃/min) to 160℃ for 1 min. Nitrogen gas was used as carrier and FID as detector. The flow of carrier was 1.0 ml/min，the temperature of injection port and detector was 200℃ and 250℃, respectively.
Results： The limits of detection for acetone，methanol，dichloromethane，ethyl acetate，isobutanol，pyridine，toluene in 7-ACCA were 2.5 μg/ml，1.5 μg/ml，15 μg/ml，2.5 μg/ml，2.5 μg/ml，2.5 μg/ml and 11 μg/ml，respectively. Only acetone was detected in the sample，and was less than the limits of Ch.P.
Conclusion： The method can effectively detect the residual solvents in 7-ACCA.

Objective： To investigate the preparation method of polystyrene core-poly (acrylamide-acrylic acid) shell fluorescent microspheres.
Methods： The polystyrene core-poly (acrylamide-acrylic acid) shell (P-(St-co-AAM)) fluorescent microspheres were prepared using fluorescent microspheres as the core and acrylamide/acrylic as polymerization monomer. Reaction conditions affecting the morphology of core-shell structure including feeding mode，initiator，cross linker，pH，concentration and swelling were studied.
Results： Fluorescent microscopy showed that the relatively uniform particle sizes were distributed in a range of 7-8 μm. Fourier transform infra-red spectroscopy（FT-IR）proved the existence of poly (acrylamide-acrylic acid) shell and amide group on the surface. The optimal conditions for seeding polymerization: azobisisobutyronitrile was used as the initiator in the absence of cross linker，after a 40 h swelling treatment by using alcohol with the appropriate reaction temperature (70℃)，reaction time (3 h) and pH(6-7). The average dispersion and stability were 25.14% and 90.21%,respectively. The fluorescein release percentage was kept stable at approximately 30% after 40 h.
Conclusion： The fluorescent microspheres prepared by this method have core-shell structure and satisfactory fluorescence properties with good dispersion and stability.

Objective： To investigate the effect of benzo(a)pyrene (BaP) on platelet aggregation and expression of P-selectin.
Methods： Blood samples were collected from healthy volunteers and the platelets was washed. Platelet aggregation was monitored by aggregometer and the expression of P-seletin was detected by whole blood flow cytometry.
Results： BaP (10 μmol/L，1 μmol/L and 0.1 μmol/L) did not induce platelet aggregation; however，preincubation with BaP (10 μmol/L) significantly enhanced ADP-induced platelet aggregation (P<0.01) and platelet aggregation was (80±10)%，while BaP-preincubation failed to enhance platelet aggregation under collagen and thrombin stimulation. Flow cytometry showed that preincubation with BaP increased ADP- induced，but not thrombin-induced P-selectin expression (P<0.01).
Conclusion： BaP can stimulate ADP-induced platelet aggregation and P-selectin expression，probably through the interaction with ADP-mediated signal pathway.

Objective： To investigate the effect of small interfering RNA on cell proliferation and apoptosis in gastric cancer cell lines with high expression of RegIα.
Methods： Total RNA was isolated from six gastric cancer cell lines,and the expression of RegIα mRNA was detected by RT-PCR. RegIα RNAi expression vector was constructed and stably transfected into MKN45 and AGS cells with high RegIα expression,empty-vector was used as control. RegIα mRNA and protein expression was measured by RT-PCR and Western blot respectively in stable transfected cell lines. Cell proliferation and apoptosis were detected with MTT assay and flow cytometry.
Results： RT-PCR results indicated that RegIα mRNA expression was significantly inhibited by RNAi in both cell lines compared with empty-vector. Western blot results showed that RegIα protein was down-regulated to (44±4)% and (25±4)% respectively in MKN45 and AGS cells compared to empty-vector. MTT results showed that cell growth was significantly inhibited in MKN45 and AGS cells.The apoptosis rate in MKN45 and AGS cells was remarkable increased compared to that of empty-vector (12.96±0.50)% vs (3.99±0.30)% and (11.59±1.10)% vs (4.22±0.40)%(P<0.05).
Conclusion： Small interfering RNA of RegIα gene can efficiently down-regulate RegIα expression in MKN45 and AGS cell lines,and further inhibit cell growth and induce cell apoptosis.

Objective： To investigate the effects of atorvastatin on matrix metalloproteinase-9 (MMP-9) and the tissue inhibitor-1 of matrix metalloproteinase (TIMP-1) levels in bronchoalvcolar lavage fluid (BALF) and serum of rats with experimental pulmonary fibrosis.
Methods： Pulmonary fibrosis was induced by intratracheal administration of bleomycin in 30 female rats,which were further divided into two groups:Group M (without treatment) and Group A (treated with atorvastatin 10 mg/kg); control group (n=5,Group C) was intratracheally administrated with same volume of saline. Five animals were sacrificed at 2 weeks (M2 and A2),4 weeks (M4 and A4) and 6 weeks (M6 and A6) after model establishment,respectively. Lung tissue samples were harvested and prepared for HE and Masson′s trichrome staining. Concentrations of MMP-9 and TIMP-1 in BALF and serum were measured by ELISA.
Results： The severity of inflammation and pulmonary fibrosis was significantly reduced in Group A than that in Group M,especially at week 6. No significant difference was noted in the serum concentrations of MMP-9 and TIMP-1 among the Group M,A and Group C. The BALF concentrations of MMP-9 in Group M2 and M6 were significantly higher than those in Group C（P<0.01 and 0.05）,whereas those in the atorvastatin groups (A2,A4 and A6) were lower than those in M2,M4 and M6. Although the MMP-9 was still higher in Groups A2 and A4 than in the Group C,there was no significant difference in MMP-9 between Group A6 and Group C. TIMP-1 levels in BALF were significantly higher in M4 and M6 than Group C (P<0.01 and 0.05),there were no significant differences between Group M2 and Group C. The TIMP-1 levels in BALF of atorvastatin groups were significantly lower than those of model groups and control group（P<0.01 and 0.05）,which resulted in a significantly increased ratio of MMP-9 to TIMP-1 in the atorvastatin groups.
Conclusion： Atorvastatin inhibits the synthesis and release of MMP-9 and TIMP-1 in the lung tissue of rats with bleomycin-induced pulmonary fibrosis,and has no significant effect on circulating MMP-9 and TIMP-1,which may be associated with the attenuation of experimental pulmonary fibrosis in rats.

Objective： To investigate the effect of AT1 receptor on the changes of thyrosine hydroxylase-immunoreactivity (TH-IR) in rostral ventrolateral medulla (RVLM) induced by brain cholinergic stimuli in rats.
Methods： Male SD rats were randomly divided into 4 groups: NS+CBC group，Los+CBC group，Los+NS group and NS+NS group. AT1 was blocked by pretreatment of 20 μg losartan in Los+CBC and Los+NS groups; intracerebroventricular injection of 0.5 μg carbachol was used for cholinergic stimuli in NS+CBC and Los+CBC groups; normal saline (NS) was used for control. The output amount of natrium in kidney，glomerular filtration rate (GFR) and renal plasma flow (PRF) were observed. The changes of TH-IR in the RVLM were observed by immunohistochemistry.
Results： In NS+CBC group carbachol induced potent natriuresis，after pretreatment of losartan the natriuretic effect was partially inhibited in Los+CBC group. Both the number and optical density of TH-IR positive neurons in NS+CBC group were markedly increased than those in NS+NS group(P<0.05); while those in Los+CBC group were significantly lower than those in NS+CBC group(P<0.05). Intracerebroventricular injection of carbachol and losartan had no effect on GFR and RPF(P>0.05).
Conclusion： The results suggest that cholinergic stimuli can induce potent natriuresis and increase the activity of adrenergic neurons in the RVLM; the above effects can be down regulated by blockade of brain AT1 receptor.

Objective： To evaluate the clinical outcome among three different laser in situ keratomileusis （LASIK） ablations: Q-factor customized ablation（aberration smart ablation ,ASA)，wave-front guided ablation (WASCA) and ablation under wave-front guiding plus iris recognition system (IR+WASCA).
Methods： This prospective study comprised 96 eyes of 96 patients，and they were randomly divided into three groups: 30 patients in ASA group，32 in WASCA group，and 34 in IR+WASCA group. There were no any statistical differences in spherical equivalent（SE），age，sex，pupil diameter，higher-order aberrations (HOA) among three groups preoperatively. Wave-front analysis was performed before and 1，3 months after operation.
Results： All patients got an uncorrected visual acuity (UCVA) ≥0.8 1 and 3 months after operation. The residual SE was in ±0.50D both at 1 and 3 months after surgery. There was no statistical difference in SE value，HOA，change of HOA，and coma aberration postoperatively among three groups. Horizontal coma (Z31) aberration took the majority of coma. HOA，total coma aberrations and spherical aberration increased postoperatively (P=0.000)，but without significant difference between 1 month and 3 months after surgery.
Conclusion： That three customized LASIK are all effective，safe，accurate and stable; meanwhile WASCA may have better UCVA than the other two groups postoperatively.

Objective： To investigate the sleep quality and influencing factors of perimenopausal womem in Ningbo region.
Methods： A total of 527 perimenopausal women who lived in Ningbo for more than 5 years were enrolled in the study. The subjects were surveyed by self-designed questionnaire and Pittsburgh sleep quality index (PSQI); PSQI >7 was defined as the cut-off value for sleep quality. Data were analyzed by software SPSS 11.0.
Results： The mean PSQI score was 5.79±3.08 and 23.6% of perimenopausal women in the study showed poor sleep quality. The proportion of poor sleep in 55-60 age group was 40.2%，significantly higher than that of 40-44，45-49 and 50-54 age groups (P<0.05). The sleep quality was associated with physical exercise，social activity，sleep circumstance，anxiety and family income in perimenopausal women.
Conclusion： The sleep quality of perimenopausal women and the influencing factors are associated with mental health, life style and sociol environment, which suggests that some interventions should be undertaken to improve the sleep quality of perimenopausal women.

Objective： To evaluate the combination of drop in canales sacralis with acupotomy dissolution in the treatment of lumbocrural pain caused by slipped discs.
Methods： One hundred and thirty-nine patients with lumbocrural pain caused by slipped discs were randomly divided into 3 groups:cases in Group A were treated by the drop in canales sacralis,in Group B by acupotomy dissolution and in Group C by the combination of canales sacralis drop with acupotomy dissolution. MacNab score and VAS score were assayed before treatment and 1 week,3 and 6 months after treatment.
Results： The effective rates in Groups A,B and C at 1 week, 3 and 6 months after treatment were 71.4%,75.5%,79.6%; 75.0%,79.6%,81.8% and 89.1%,91.3%,93.5%,respectively (P<0.01). The pain intensity in Group C was reduced more markedly at different time points after treatment than that in Group A and Group B (P<0.01).
Conclusion： The combination of canales sacralis drop with acupotomy dissolution is superior to each method used alone in treatment of lumbocrural pain caused by slipped discs in the short- and long-term.

Primary and secondary corneal endothelial decompensation leads to stromal edema，corneal opacity and loss of visual acuity. The pathogenesis of corneal endothelial decompensation is that adult corneal endothelium in vivo lacks of a robust proliferative response to injury，does not divide sufficiently to replace the lost cells. Previous studies indicate that cell-cell contact inhibition and transforming growth factor-beta2 (TGF-β2) in aqueous humor may be responsible for maintaining human endothelial cells in a non-replicative state in vivo. The results of the experimental investigation by using immunofluorescent staining of the cell cycle-associated proteins and cell proliferation marker Ki67 in corneal endothelium indicate that human corneal endothelial cells in vivo are arrested in the G1-phase and have not exited from the cell cycle. Successful outgrowth in culture of human corneal endothelial cells in vitro and the establishment of the immortalized human endothelial cell line，provide strong evidence that corneal endothelial cells retain proliferative capacity. Experiments with cell culture ex vivo demonstrate that corneal endothelial cells cultured from young donors grow more robustly than those from older donors，and cells cultured from peripheral area of corneas show greater cell density than central regions. Studies have demonstrated that in vitro human corneal endothelia undergo mitotic changes in response to stimulation of growth promoting agents，such as growth factors，EDTA and extracelluar matrix. Identification of corneal endothelial stem cells and isolation and culture of human endothelial precursor cells in vitro will be beneficial for further investigation regarding the mechanism of corneal endothelial regeneration as well as corneal endothelial cells in vitro culture.

Inert gas is a group of rare gases with very low activity，their application in medical field has increasingly drawn attentions. It is known that inert gases helium，xenon and argon have protective effects on nervous system and the mechanisms are related to eradicating free radicals，anti-inflammation，suppressing apoptosis，influencing ion channels and so on. Further study on the neuroprotective effect of inert gas will shed light on a new approach to treat neurological diseases．

LMO4 is a novel member of the LIM-only (LMO) subfamily of LIM domain-containing transcription factors,so named because they are composed almost entirely of two tandem LIM domains.This subgroup of LIM proteins has 4 members:LMO-1,LMO-2,LMO-3 and LMO-4.They all play important roles in the normal mammalian development,functioning as an important regulator of cell proliferation.LMO4 is highly expressed in the epithelial compartments at locations of active epithelial-mesenchymal interactions,and can interact with some signaling pathways involved in epithelial-mesenchymal signaling.Thus the disregulation of LMO4 expression may be involved in tumorigenesis.In this paper,we will at first expound LMO4 in detail,based on which the possible mechanisms for its interaction with TGF-β signaling and the roles of this cross-talk between them in the vital process of cell will be introduced.All of those will add to our understanding of tumerigenesis and contribute to the search of new targets for the treatment of cancer.