Objective To study the transformation mechanism of triterpenes in processing of Alisma orientalis. Methods The triterpene transformations of A. orientalis pre and post-processing were comparatively analyzed by techniques of HPLC and Packed Column Supercritical Fluid Chromatography (SFC). Results In baked processing (70 ℃) of A. orientalis, little alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B.However, more alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B, then both of them were further transformed into alisol A in processing under high temperature (160-200 ℃). Conclusion Transformation of alisol B 23-acetate has two routes when A. orientalis is processed under high temperature: For one, alisol B 23-acetate is rearranged into alisol A 24-acetate which could be deacetylated into alisol A; for the other; it is deacetylated into alisol B first, then transformed into alisol A.

Autophagy is a metabolic pathway that widely presents in eukaryotic cells through the lysosomal mechanism to degrade its components. Autophagy regulates cell death not only by activating classic autophagosomal-lysosomal pathway, but also by influencing the occurrence and development of apoptosis and necrosis. Currently, the effect of autophagy in neuronal injury after acute cerebral ischemia/hypoxia and its specific mechanisms remain unclear. Studies have demonstrated that the autophagy after ischemia/hypoxia has a neuroprotective effect, such as maintaining neuronal homeostasis and reducing neuronal death; but other studies have also suggested that autophagy may aggravate neuronal injury after ischemia/hypoxia by activating multiple pathways, and even induce neuronal death.

Objective To evaluate catheterization in pancreatic duct before endoscopic papillary bal-loon dilation (EPBD)to prevent pancreatitis after EPBD.Methods Forty-three patients with normal serum amylase levels,diagnosed as having bile duct stones,underwent EPBD.Twenty-three were assigned to experi-mental group,where catheters(ERCP imaging tube)were placed in pancreatic duct before EPBD,then the pa-pillary balloon was expanded to 10 mm.Twenty were assigned to control group where eight-millimeter-diameter papillary balloon was used to remove the stones.The serum amylase levels before EPBD,6 hours and 24 hours after EPBD,the incidence of pancreatitis and high serum amylase levels associated with EPBD,as well as the mean time and success rate of removing the stones of the two groups were compared.Results Post-EPBD pan-creatitis occurred in one patient in experimental group (4.35%),and seven in control group (35.00%), which was significantly different(P <0.05).Meanwhile,the mean levels of serum amylase 6 h and 24 h after EPBD in the experimental group were (102.61 ±98.99)U /L and (60.35 ±26.18)U /L respectively,lower than those in the control group (398.25 ±259.32)U /L and (230.50 ±281.31)U /L(P <0.05).After the papillary balloon was expanded to 10 mm in experimental group,the mean time of removing stones was (10.43 ±2.27)min,which was shorter than that of control group (17.90 ±4.49)min (P <0.05).Stone-re-moving rate of two groups had no difference and they all succeeded one time.Conclusion Placing catheter in pancreatic duct before EPBD to prevent pancreatitis after EPBD makes it easier to remove stones in shorter op-eration time.It can prevent pancreatitis and high amylase blood disease after EPBD.

Objective To study any functional changes in the trigeminal nociceptive system in cases of migraine with aura (MWA) using painful heat-evoked potentials (CHEPs),and to explore the value of CHEP in evaluating the trigeminal small nerve fibers of patients with MWA.Methods Sixty patients with MWA were recruited as the MWA group,and 60 healthy persons were selected as a normal control group.Thermal stimulation at 51 ℃ was applied to the skin above the patients' right and left eyebrows.The N wave latencies and N-P amplitudes of the CHEPs were recorded at the vertex (Cz) of the head.The intensity of the stimulus was graded using a visual analogue scale (VAS).Results In the control group the simulation produced no significant differences in the N550 or P750 latencies or in the N-P amplitudes.In the MWA group no significant differences were observed when the headache side was stimulated,but the N550 and P750 latencies were both significantly shorter than those of the control group and the N-P amplitude was significantly greater than that of the control group.The average VAS score on both sides in the patients with migraine was significantly higher than those of the control group.However,there was no significant difference in average VAS scores between the headache and non-headache sides of the patients with migraine.Conclusions CHEP is a reliable method for detecting any pathological changes in the trigeminal small nerve sensory pathway.Increased cutaneous allodynia in acute migraine attacks is detectable using CHEPs.

Objective To optimize the concentrate process ofDanshen injection.Methods The concentration changes of Salvianic acid A sodium, Protocatechuic aldehyde and Salvianolic acid B were selected as indexes to compare mospheric concentrate, vacuum concentrate, nanofiltration concentrate and nanofiltration-vacuum coupling concentrate methods with HPLC. Results Phenolic acid composition changed when atmospheric and vacuum concentrate method were used. Retention rate of three phenolic acids were over 93% after nanofiltration concentration, but the concentration efficiency was limited by solution viscosity. Nanofiltration-vacuum coupling concentrate methods could solve the above problems, meanwhile, retention rate of three phenolic acids were over 91%.Conclusions Nanofiltration technology takes advantage in medicine extraction concentrate with heat sensitive ingredients, and its application prospect in traditional Chinese medicine pharmaceutical industry would be broad.

Objective To analyze the association between mtDNA mutations and photodamagc after ultraviolet B (UVB) irradiation. Methods Primary human skin fibroblasts (HSF) and primary human epi- dermal keratinocytes of adult (HEKa) were irradiated by sub-lethal doses of UVB thrice a day for 4-5 days. Thereafter, genomic DNA was extracted from irradiated cells and conventional PCR was applied to detect the frequency rates of 4977 bp and 3895 bp mtDNA deletion. To quantitatively analyze the mutation levels, SYBR Green real-time PCR method was performed. Results In both cell lines, the frequency rates and relative copy number of deletions increased with the cumulative doses of UVB exposure (P<0.05). The prevalence rate of 3895 bp deletion peaked 53.3% and and relative copy number reached (49.63±4.38)×10-5, showing a more intense response to the accumulation of UVB radiation than 4977 bp deletion. In HSF, the minimum cumu- lative dose of UVB radiation was 150 mJ/cm2 for the induction of 3895 bp deletion, and 200 mJ/cm2 for the induction of 4977 bp deletion. It seemed that mtDNA deletion was more readily to be induced by UVB radia- tion in HSF than in HEKa. Conclusions The development and accumulation of mtDNA mutation are intimately related with cumulated UVB dose received by skin cells, and the 3895 bp deletion is more reliable in moni- toring the photodamage caused by UV than 4977 bp deletion. Therefore, the 3895 bp deletion may serve as a biomarker for the detection of photodamagc in skin cells. HSF appear to have an increased susceptibility to UVB radiation, which results in a higher frequency and level of mtDNA mutations compared with HEKa.

Object To optimize the preparation procedure for Zhixuan Granula (ZXG). Methods The optimum extracting conditions of ZXG were selected by orthogonal test with the active components: 23-acetate alisol B, atractylenolide I, and dried extract as the index, it mice sedation of ZXG was clarified by pharmacodynamics. Results The optimum preparation procedure was as follows: Rhizoma Alismatis and Rhizoma Atractylodis Macrocephalae were extracted with alcohol first, adding 12-fold 70% alcohol by refluxing, extracting twice, 2 h once, then extracted with water, adding 14-fold water, extracting twice, 2 h once. The extract showed the obvious effect on sedation of mice. Conclusion The optimum preparation procedure is reliable, with higher extracting ratio of the active components.

Objective: The quality standards for Zhixuan Granule (Rhizoma Alismatis, Rhizoma Atractylodis Macrocephalae, etc.) were studied. Methods: The TLC methods for identification of Rhizoma Alismatis、 Rhizoma Atractylodis Macrocephalae were established. A simple HPLC was established for the determination of 23-acetate alisol B. The mobile phase was acetonitrile-water(70∶30). UV detecting wavelength was at 208nm. Results: Rhizoma Alismatis and Rhizoma Atractylodis Macrocephala could be detected. 23-acetate alisol B showed a linear relationship at the concentration range of 99～1388.8ng, r=0.9999. The average recovery was 103.05% and RSD was 2.41%(n=6). Conclusion: This method is suitable for the quality control of Zhixuan Granule.

Objective To study the pharmacokinetic features of silibinin from Jiqi Injection(JI) in Beagle dogs,and to observe the effect of Panax notoginseng saponins,another active component in JI,on pharmacokinetics of silibinin. Methods The Beagle dogs received intravenous injection of JI and silibinin,and then its plasma sample was collected in different time. The plasma samples of Beagle were prepared by hydrolysis with sulfatase-? glucuronidase complex enzyme and liquid-liquid extraction with aether. A high-performance liquid chromatographic method was developed to determine the plasma concentration of silibinin,and the pharmacokinetic parameters were performed by BAPP2.3 program. Results The pharmacokinetics of two tested preparations met with two-compartment model. There were not significant differences between pharmacokinetic parameters of JI and that of Silibinin Injection. Conclusion The silibinin in Jiqi Injection has a fast in-vivo clearance rate after intravenous injection,and Panax notoginseng saponins have no effect on its pharmacokinetic parameters.