Object To establish a HPLC method for determination of ethyl p hydroxybenzoate in Dragon's Blood. Methods Using C 18 reverse phase column and acetonitrile 1% acetic acid (31∶69) as mobile phase, detecting at 257 nm and quantitating with external standard method. Results The standard curves of ethyl p hydroxybenzoate showed good linearity over the range of 0.206-2.884 ng, r=0.999 8. The average recovery was 96.02% and RSD was 1.87%. Conclusion The method is quick and simple, and it is accurate and reliable for the determination of ethyl p hydroxybenzoate in Dragon's Blood.

Objective: To establish HPLC method for the determination of loureirin A and loureirin B in Sanguis Draxonis Capsules.Methods: HPLC system included C 18 reverse phase column and acetonitrile 1% acetic acid (34∶66) as mobile phase, detection at 280nm and external standard method.Results:The standard curves of loureirin A was linear in the concentration range of 98～490ng, r =0.9996.The average recovery was 97.45%, RSD was 1.82%.The standard curves of loureirin B was linear in the concentration range of 43.2～259.2, r = 0.9993 .The average recovery was 96.92%, RSD was 1.57%.The contents of loureirin A and loureirin B in Sanguis Draxonis from heating free technology were higher than traditional technology.Conclusion: This heating free technology is a new technique that is a creation with higher extract rate,this method is worth populariging.

Objective · To design and synthesize a series of benzenesulfonamide derivatives, test their inhibitory activity to CDK2-cyclinA2 kinase, and investigate the structure-activity relationship. Methods · Virtual screening was executed via computer-aided drug design according to the ATP binding site in CDK2-cyclinA2 protein crystal. A series of benzenesulfonamide derivatives were designed and synthesized on the basis of the interaction modes between the lead compound and the CDK2-cyclinA2. The biological evaluation of compounds was made through the CDK2-cyclinA2 in-vitro kinase activity detection system. Results · Twenty-nine new benzenesulfonamide compounds were prepared, and their inhibitory activity to CDK2-cyclinA2 was elicited. WZ-026 had the highest inhibitory parameter, which half maximal inhibitory concentration (IC50) was 3.81 μmol/L. Conclusion · By multipurpose utilization of virtual screening, chemical synthesis, and biological activity test, a benzenesulfonamide compound WZ-026 was found, which has great inhibitory activity towards CDK2-cyclinA2. Preliminary structure-activity relationship of compounds was obtained.

Object To identify the differences of chemical constituents of Dragon's Blood obtained by different extract processing. Methods The mentioned Dragon's Blood was analyzed by TLC, UV spectro photometry, and HPLC. Results The contents of dihydrochalcone in Dragon's Blood by the new extract processing were higher than that by traditional one's, while the contents of flavonoid and stibene were lower. Conclusion The heating free technology is a new technique that is an original with higher extract rate, and it is worth of spreading. It will promote the modernization of TCM.

Objective: To establish HPLC method for determination of pterostilbene in Dragon's Blood with different extraction technology. Method: Applying a C 18 phase column and acetonitrile 1% acetic acid(41∶59) as mobile phase, detecting at 319nm and quantitating with external standard method. Results: The standard curves of pterostilbene was linear in the concentration range of 10.4～104ng, r =0.9992. The average recovery was 98.28%, RSD =1.97%. The content of pterostilbene in Dragon's Blood extracted with ordinary temperature was lower than that with heating technology. Conclusion: The content of pterostilbene extracted with ordinary temperature extraction in Dragon's blood have the advantage of heating extraction.

BACKGROUND: Based on our previous researches in mechanism studies and clinical applications of human hair keratin (HHK), a new concept "in vivol in situ tissue engineering" has been proposed. Under the guidance of this theory, a scaffold of HHK-collagan sponge (inner layer) combined with poly (2-hydroxyethyl methacrylate) (PHEMA) (outer layer as a drug delivery carrier) would be developed to investigate its feasibility to be as a dermal dressing. OBJECTIVE: To develop a scaffold composed of HHK-collagan sponge (inner layer) combined with PHEMA film containing polydatin(PD)(outer layer as a drug delivery carrier) and to evaluate the therapeutic efficacy of the HHK-collagen sponge-PHEMA/PD complex on burn wound healing. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Department of Histology and Embryology, Southern Medical University between March and December 2005. MATERIALS: Burn was induced in 15 male Sprague-Dawiey (SD) rats, Rat models of burn were evenly randomized to 3 groups: experimental, positive control, and negative control. METHODS: ①HHK-collagen sponge was prepared through combination of a HHK meshwork (1mm × 1 mm in size for each grid) made up of three components (determined according to biochemical procedures of various degrees, i.e., light, medial, and severe) at a ratio of 4:3:3 with primary collagen sponge extracted from bovine tendons in a mould. Sponge film (used as inner layer dressing) was made by vacuum freeze-drying. ② PHEMA was prepared by polymerization. Than PD was added to prepare PHEMNPD film (used as outer layer dressing).③ Degree Ⅱ burn wound models were established in SD rats by scalding, Superficial necrotic tissue was removed from burn wounds at postnatal 3 days and leave the denatured dermis remained. The wounds were either covered with human HHK-collagen- PHEMNPD complex in the experimental group, or with glutaraldehyde-treated porcine skin in the positive control group, and sterile absorbent gauze was used in the negative control group. MAIN OUTCOME MEASURES: ① Complete epithelization was taken as the standards, and at postoperative 7, 14, and 21 days, wound healing was respectively calculated. ② At postoperative 1, 2, 4, 6, and 8 weeks, the whole wound surface and its peripheral tissue were dissected for observing granulation tissue growing under an optical microscope and detecting the collagen fiber and elastic fiber in the newly formed tissue by immunohistochemical staining. RESULTS: ① Gross observation results revealed that in the experimental group, the volume of the diffusate under the ideal moisture was less compared with the positive control group; the healing time was slightly shorter in both the experimental group and the positive control group than in the negative control group (P= 0.000); At postoperative 7, 14, and 21 days, the healing rate was higher in the experimental and positive control groups than in the negative control group (P=0.000), in addition, the experimental group exhibited higher healing rate than the positive control group at postoperative 14 days ( P ＜ 0.05). ②Optical microscope results showed that at postoperative 2 weeks, a small quantity of collagen fibers were found in the wound granulation tissue in all 3 groups, in particular in the experimental group. Immunohistochemical staining results regarding collagen protein and elastin revealed that at postoperative 2 weeks, both the fine strip-like type Ⅰ collagen fibers and a few silk-like elastic fibers were stained yellowish-brown in the dermal matrix in the experimental group, which were weakly positive in the positive control group, while there was no elastin detectable in the negative control group; at postoperative 8 weeks, burn wounds in all the 3 groups werefully recovered. Remodeling of collagen fibers was more obvious in the experimental and positive control groups than in thenegative control group, while the tendency to scar formation with derangement of epithelial cells and collagen fibers in dermis was more prominent in the negative control group than in the remaining two groups.CONCLUSION: HHK-collagen sponge-PHEMA/PD complex may be a new burn dressing via in vivo construction of tissueengineered epidermis, in which PHEMA may be a feasible drug-delivery carrier.

Objective To explore whether the peritoneal cooling was better than other cooling methods on protection neuron damage of the hippocampus CA1 after cardiopulmonary resuscitation (CPR) in New Zealand rabbits.Methods Forty eight adult New Zealand rabbits were induced ventricular fibrillation by AC current and were resuscitated after cardiac arrest for 5 minutes.The rabbits were randomly divided into four groups according to the way of cooling methods,nomothermia group ( NT),peritoneal cooling group (PC),surface cooling group (SC) and local cooling group (LC).The changes of tympanic membrane temperature were recorded in each animal and blood plasma concentrations of electrolyte were tested in each group at different time points after restore of spontaneous circulation (ROSC).Brain tissue were removed,the numbers of vigorous and apoptotic neurons in the hippocampus CA1 area were counted after ROSC at 72h.One-way ANOVA or Mann-Whitney rank was used to determine the statistical significance between two groups.LSD-t test for multiple comparisons,R × C test for ROSC comparisons,a two-tailed value of P ＜0.05 was considered statistically significant. Results Hypothermia was rapidly induced in PC after ROSC,and the time of arriving at target temperature was (26 ±7) min in PC,(60 ±9) min in SC,(69 ± 12) min in LC respectively; in the maintain hypothermia period,the tympanic membrane temperature was maintained at 33～ 35 ℃ in each group exception nomothermia group (NT).There were no differences with main electrolyte,acid-abase liquid balance and renal function between each group at each time point after ROSC.The numbers of vigorous neurons in hippocampus CA1 area were ( 37.07 ± 6.43 ) /40 × in NT group,(35.13 ± 6.97) /40 × in LC group,(55.76 ± 10.13 ) /40 × in PC group,and (50.70 ± 7.38 ) /40 × in SC group (PC:NT,P＜0.01,SC:NT,P＜0.01,PC:SC,P=0.043,PC:LC,P＜0.01,LC:NT,P=0.52).The numbers of apoptotic neurons were (44.07 ±6.09) /40 × in NT group,(29.88 ±4.81 ) /40× in PC group,( 33.55 ± 5.67 ) /40 × in SC group and ( 42.27 ± 5.20 ) /40 × in LC group respectively (PC:NT,P ＜0.01,SC:NT,P ＜0.01,PC:LC,P ＜0.01,SC:LC,P ＜0.01,PC:SC,P=0.026,LC:NT,P =0.364 ).Conclusions The new peritoneal cooling method could rapidly induce and maintain hypothermia,and it had better protections on neurons in hippocampus CA1 than surface cooling and local cooling method after ROSC in New Zealand rabbits.

Objective To examine the therapeutic effect of C Ⅱ TA inhibition in collagen-induced arthritis(CIA),using a delivery system tailored to target C ⅡTA gene by small interfering RNA (siRNA).Methods Mice with collagen-induced arthritis were injected intravenously with C Ⅱ TA siRNA.The clinical score was monitored for up to 4 weeks after treatment.The severity of inflammation of mouse joint was evaluated by histological examination.Real-time PCR was used to determine the cytokine mRNA expression.Cytokine production was measured by ELISA from serum.T cell proliferation was examined by MTT method.Results IFN-γ and IL-17 were elevated in CIA mice,but were iuhibited significantly by C Ⅱ TA siRNA either prevention or intervention of autoimmune arthritis.Collagen specific T cell proliferation was significantly suppressed.Increased level of IL-4 by T cells was observed in C Ⅱ TA siRNA treated group compared with that of control group.Conclusion Our findings indicate that systemic RNAi-mediated C Ⅱ TA gene silencing is effective in the treatment of CIA and regulateds the balance of Th1/Th2 differentiation.

Objective To evaluate the effect of percutaneous coronary intervention (PCI) or thrombolysis,in patients with return of spontaneous circulation (ROSC) after out-of-hospital cardiac arrest (OHCA),in the presence of ST-elevation myocardial infarction (STEMI).We demonstrated the benefits of the two therapies on ROSC patients in hospital discharge and neurological recovery,and clarified the importance of ROSC,so as to guide the treatments for OHCA in the presence of STEMI.Methods It was performed a meta-analysis of clinical studies located in PUBMED and MEDLINE databases from January 1995 to October 2011.OHCA patients with ROSC were as our study objects,the hospital discharge and neurological recovery rates,of patients with and without PCI or thrombolysis,were assessed in patients with ROSC after OHCA in the presence of STEMI.In the same Cohort Study,between received and rejected PCI,or between received and rejected thrombolysis in OHCA patients with ROSC as treated group and control group,using Review Manager 5.1 software to analyze,respectively.Furthermore,we also compared the differences in hospital discharge and neurological recovery rates between patient groups who received PCI or thrombolysis by Pearson x2 analysis.Results The meta-analysis showed that the rate of hospital discharge improved with both PCI (odds ratio [OR],1.65 ; 95％ confidence interval [CI],1.05-2.59,P ＜ 0.01)and thrombolysis (OR,2.03 ; 95％ CI,1.24-3.34,P ＜ 0.01) in patients with ROSC after OHCA,in the presence of STEMI.We also found that there were not significant differences between with PCI and with thrombolysis in the rate of hospital discharge (63.00％ vs.65.19％,P =0.548) and neurological recovery (88.62％ vs.91.25％,P =0.351) for the patients with ROSC after OHCA (P ＞0.05).Conclusions In patients with ROSC after OHCA in the presence of STEMI,both PCI and thrombolysis improved hospital discharge rates.Furthermore,there were similar efficacy in hospital discharge and neurological recovery rates between with PCI and with thrombolysis.

Objective To investigate the therapeutic effects of enhanced external counterpulsation (EECP) on cerebral edema and cerebral blood flow perfusion with MRI following cardiac arrest (CA) and on successful return of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in dogs.Methods Sixteen beagle dogs were induced CA with alternating current on epicardium,then were randomly (random number) divided into the EECP and control group after successful ROSC.MR scanning brain of all animals was carried out by diffusion-weighted imaging (DWI) and perfusion weighted imaging (PWI) before CA and on the first,second and third days after ROSC.Blood pressure,right common carotid artery flow,and intracranial microcirculation perfusion were measured.Results There were no significant differences in mean artery pressure at all intervals between two groups (P ＞ 0.05).There was significant increase in right common carotid artery blood flow and intracranial microcirculation of dogs in EECP group compared with the control group (P ＜ 0.05).Apparent diffusion coefficients (ADC) of water molecule on the first and third days after ROSC were significantly higher in the EECP group than those in the control group (P ＜ 0.05).Ratios of post-ROSC relative cerebral blood flow (RCBF) /original cerebral blood flow were higher in the EECP group than those in the control group on the first,second and third days after ROSC (P ＜ 0.05).Conclusions EECP treatment could improve cerebral blood flow perfusion and relieve ischemic cerebral edema,alleviating brain injury in dogs following CA and successful ROSC.