OBJECTIVE:To study the inhibitory effects of minocycline (MC) on TNF-α induced monocyte-endothelial adhe-sion and the relative mechanism. METHODS:Primary human umbilical vein endothelial cells (HUVECs) were isolated from hu-man umbilical veins with enzyme digestion. HUVECs were divided into blank control group,model group,MC low-dose,medi-um-dose and high-dose groups(1,10,100 μmol/L). After treated for 2 h,10 ng/ml TNF-α was employed to stimulate monocytes THP-1 adhesion with HUVECs except for blank control group,in order to induce monocyte-endothelial adhesion model. The num-ber of adherent cell was observed by fluorescence microscope,and fluorescence intensity was detected by microplate reader. Flow cytometry was adopted to detect the expression of intercellular adhesion molecule (ICAM)-1. The expressions of NF-κB p65 pro-tein in cell nucleus and cytoplasm were detected by Western blot. RESULTS:Compared with blank control group,the number and fluorescence intensity of adherent cell,the expression of ICAM-1 and the protein expression of NF-κB p65 in nucleus were all in-creased in model group,while the protein expression of NF-κB p65 in cytoplasm was weakened,with statistical significance(P<0.01). Compared with model group,the number and fluorescence intensity of adherent cell,the expression of ICAM-1 were all de-creased in MC low-dose,medium-dose and high-dose groups;the protein expression of NF-κB p65 in nucleus was weakened in MC medium-dose and high-dose groups,while the protein expression of NF-κB p65 in cytoplasm was heightened,with statistical significance (P<0.01 or P<0.05). CONCLUSIONS:MC can inhibit TNF-α induced monocyte-endothelial adhesion by a likely mechanism of reducing the expression of ICAM-1 in HUVECs and inhibiting the expression of NF-κB p65 protein.

Objective In order to study the application of macroporous resins and so on to the purified active components of Euphorbia humifusa、Leaves of Flos Lonicerae、Chrysanthemum morifolium,adsorption and separation properties for 3 types of macroporous resins and polyamide were investigated.Methods The total flavone was used as the evaluating criteria,we selected suitable macroporous resins and studied optimum technological parameters of the adsorption and elution.Spectrophotometry was used for the determination of total flavone.Results The suitable macroporous resins which were used to the purified active components of traditional Chinese medicine were D101 and DA201 and DM301 for Euphorbia humifusa、DA201 for Leaves of Flos Lonicerae、D101 and DA201 and DM301 for Chrysanthemum morifolium,The concentration of the sample of Euphorbia humifusa for DA201 and D101 were 0.49～1.47 and 0.42～1.31 mg/ml.The concentration of the sample of Leaves of Flos Lonicerae for DA201 was 1.03～2.07 mg/ml.The concentration of the sample of Chrysanthemum morifolium for DA201 and D101 was 0.50～1.00 and 0.71～1.99 mg.ml.In the adsorption course,appeared leaking were 8 and 10、2、2 and 1 BV respectively.In the elution course,when the alcohol concentrations were 20%、30%、40% and 20%、30%、40%;10%、20%、30%;30%、40%、50% and 20%、30%、40%;respectively,the total flavone content in the elution solutions was higher.The influence of temperature to DA201 and D101 adsorpting total flavone for Euphorbia humifusa was not great.But the influence of temperature to DA201 and D101 adsorpting total flavone for Leaves of Flos Lonicerae and Chrysanthemum morifolium were certain degree.Conclusions It is obviously different to refine the total flavone active components of traditional Chinese medicine,while using 3 types of macroporous resins and polyamide.

Objective To study the influence of different dosage forms on the release rate of total salvianolic acid of sus- tained-released Danshen extract preparations.Methods With salvianolic acid B as the standard,uhraviolet spec- trophotometry and the release test were applied to determine the accumulated release rate of total salvianolic acid of sus- tained-released Danshen extract preparations(R1～R9)of 9 different dosage forms and to evaluate the release behav- ior of sustained-released Danshen extract preparations in vitro.Results The accumulated release rate of total sal- vianolic acid in R7 was the best,and the in vitro release behavior was consistent with Higuchi equation:M_1/M_∞= 0.363 4t~(1/2)-0.1656,r= 0.996 2.Conclusion R7 is the first choice dosage form for the preparation of the sus- tained-released Danshen extraction preparations.

【Objective】 To assay the naringenin content in different medicinal materials of Exocarpium Citri Grandis (ECG). 【Methods】 High performance liquid chromatography (HPLC) was applied to determine the naringenin content in different medicinal materials of ECG. The chromatographic conditions were: gradient elution being performed with methanol as mobile phase A and water - acetic acid (61: 4) as mobile phase B, flowing rate being 1.0 mL/min; detecting wavelength at 283 nm. 【Results】 In the rage of 0.088～0.880 ?g, naringenin showed a good linearity with the chromatographic peak area, r=0.9999, average recovery rate was 95.49% and RSD=2.00% (n=5). The naringenin content in 23 different samples of ECG was in the range of 0.1～1.0 mg?g~(-1). 【Conclusion】 It is the first time to establish the HPLC method for the determination of naringenin content in ECG and the method is accurate and reproducible. Naringenin content in varieties of ECG is stable and the difference of naringenin content between Citrus grandis Osbeck. Var. tomentosa Hort. and Citrus grandis (L.) Osbeck is not significant.

N-acetyltransferase 2 is an important metabolic enzyme in the human body,participating mainly in the metabolism of medicine containing nitrogen.Presently a lot of researches have been undertaken in the respects like the determination of NAT2 genotype and drug metabolism and so on.In the recent years,more and more attention is paid to the simulation of three-dimension(solid)structure of protein,but the study is just at the outset on three-dimension structure of the NAT2 as well as its structure-function relationship when combined with drug.The research progress of construction of the three-dimension structure of NAT2 is summarized,emphasizing on its methods and tools.

Objective To enable medical institutions and personnel in the case of a medical encroachment dispute to provide precise proof in court, use standard format for the legal complaint, employ the appropriate proofgathermg method, ensure the simplicity of the proof-providing procedure, and avoid the waste of manpower and material resources and the liability of economic compensations in the course of compromise settlement, mediation by the health administrative department or adjudication by the court. Methods Regulations were laid down within the hospital for proof-providing procedures, methods, contents, the contents and format of legal complaints and reply letters, and documents in preparation for the provision of proof. Results Through the handling of over 900 medical encroachment disputes, the above methods proved to be practical, applicable and up to the standard. They could save the time of the hospital, the patient and the lawyer on blind surveys, reduce proof-providing time, and improve efficiency in handling medical encroachment disputes. Conclusion Standardizing proof-providing procedures, methods, and contents and the format of legal complaints or reply letters is of great significance in improving efficiency in handling medical encroachment disputes and reducing the hospital's chances of losing lawsuits.

Objective To investigate the protective effect of desflurane preconditioning on human umbilical vein endothelial cells against anoxia-reoxygenation(A/R)injury.Methods The human umbilical vein endothelial cell line(ECV304)was provided by the Liver Cancer Institute,Zhongshan Hospital,Fudan University.ECV304 cells were randomly divided into 5 groups:group Ⅰ normal control;group Ⅱ A/R;group Ⅲ A/R+rhTNF-α;group Ⅳ Des + A/R and group Ⅴ Des + A/R + rhTNF-α.In group Ⅱ-Ⅴ the cells were exposed to 95% N2 + 5% CO2 in an incubator for 30 min followed by 60 min reoxygenation.In group Ⅲ and Ⅴ rhTNF-α(10 ng/ml)10 μl was added to the cells as soon as reoxygenation was started,while in group Ⅳ and Ⅴ the cells were pretreated with 7.2% desflurane for 30 min followed by 10 min washout before A/R.Apoptosis in endothelial cells was detected by fluorescence flow cytometry and TUNEL.Endothelial cells were examined with electron microscope for apoptosis and necrosis.Results The rates of apoptosis in the endothelial cells were significantly higher in A/R group and A/R + rhTNF-α group than in control group.Desflurane preconditioning significantly attenuated apoptosis in the endothelial cells induced by A/R and A/R + rhTNF-α respectively.Electron microscopy showed that there were significantly more necrotic cells in A/R group and A/R + rhTNF-α group.However in the two desflurane preconditioning groups(Ⅳ and Ⅴ)the cells were in a state of duplication and self-repairing.Conclusion Preconditioning with 30 min 7.2% desflurane can attenuate the A/R-induced injury to human umbilical vein endothelial cells.

Object To establish a new method to determine protocatechualdehyde in Maodongqing Sustained- release Tablet. Methods The RP- HPLC method was applied with CH3OH and 1 % acetic acid as mobile phase, flowrate at 1.0 mL/min, and detection wavelength at 312 nm. Results In the range from 11.7 ng to 140.4 ng, protocatechualdehyde was in a good line with peak area, r=0.99995, the average recovery ratio was 96.41 % , RSD=1.04 % (n=5). Conclusions It was the first time to establish the RP- HPLC method to determine and control the protocatechualdehyde in Maodongqing Sustained- release Tablet with accuracy and a good reproducibility.

Objective To investigate the efficacy,toxicity and cosmetic outcome of short-course radiotherapy with concomitant tumor bed boost after breast-conserving surgery for early stage breast cancer.Methods A total of 306 patients with T1-2 N0-1 M0 breast cancer after breast-conserving surgery were included.160 patients received whole-breast radiation to 45 Gy in 25 fractions followed by tumor bed boost of 14 Gy in 7 fractions (C group).146 patients received whole-breast radiation to 46 Gy in 23 fractions with concomitant tumor bed boost to 60 Gy in 23 fractions (S group).Kaplan-Meier method was used to calculate the local recurrence and overall survival rates and the differences were compared by Logrank test.Chi-square test was used to compared the differences of the clinical characteristics,toxicity and cosmetic outcome between the two groups.Results The follow-up rate was 100％.After a median follow up of 26 months,the 1-,2-and 3-year overall survival rates were 100％.No patient developed local recurrence.In C and S group,the incidence of grade 1 acute skin toxicity was 46.9％ and 45.1％ (x2 =0.73,P =0.695),grade 2was 16.3％ and 13.7％ (x2 =0.73,P =0.695).Grade 1 late skin and subcutaneous tissue toxicity developed in 16.9％ and 17.1％ of patients in C and S group (x2 =0.00,P =0.954).Grade 1 neutropenia occurred in 11.9％ and 13.7％ of patients in C and S group (x2 =0.23,P =0.633).In C and S group,66.2％ and 65.5％ of patients had excellent and good cosmetic outcome (x2 =0.01,P =0.927).Conclusions Short-course radiotherapy with concomitant tumor bed boost provides similar results to conventional radiotherapy in local control,toxicity and cosmetic outcome.Long-term follow up is warranted to confirm this finding.

The paper is to report the pharmacokinetics of sulfamethoxazole in healthy Han volunteers living at plain (PH) and native Han and Tibetan healthy volunteers living at high altitude (HNH and HNT). After healthy volunteers were administrated orally cotrimoxazole tablets, plasma concentration of sulfamethoxazole and metabolite N4-acetylsulfamethoxazole was determined by RP-HPLC, and plasma concentration-time data were analyzed by DAS 2.0 software to get the related pharmacokinetic parameters. The main pharmacokinetic parameters t(1/2) of sulfamethoxazole in PH, HNH and HNT were, respectively, 9.30 +/- 1.11, 10.99 +/- 1.23 and 10.44 +/- 1.05 h; tmax were 1.4 +/- 0.3, 2.0 +/- 1.1 and 1.8 +/- 0.4 h; Cmax were 94.42 +/- 15.26, 89.33 +/- 7.67 and 87.43 +/- 11.61 micro x mL(-1); AUC(0-t) were 1202.5 +/- 238.3, 1 434.7 +/- 193.9 and 1302.8 +/- 103.0 microg x h x mL(-1); AUC(0-infinity) were 1240.7 +/- 255.3, 1511.5 +/- 211.9 and 1363.9 +/- 116.5 microg x h x mL(-1); CL were 1.01 +/- 0.22, 0.81 +/- 0.12 and 0.89 +/- 0.08 L x h(-1) x kg(-1); V were 13.27 +/- 1.73, 12.81 +/- 2.15 and 13.28 +/- 1.20 L x kg(-1). Sulfamethoxazole pharmacokinetic parameters of HNH and HNT were significantly different from that of PH. The t(1/2) was significantly higher and the CL was significantly lower in HNH and HNT than that in PH, and the AUC(0-infinity) was significantly lower in HNT compared with HNH. This study found significant changes in the disposition of sulfamethoxazole under the special environment of high altitude hypoxia. This finding may provide some references for clinical rational application of sulfamethoxazole in HNH and HNT.