Objective To establish an ultra performance liquid chromatography method for the determination of 7 kinds of antibiotics(minocycline hydrochloride,ox tetracycline,tetracycline hydrochloride,chlortetracycline hydrochloride,doxycycline hydrochloride,chloramphenicol,metronidazole) in anti-acne cosmetics.Methods Seven kinds of antibiotics were separated on a Waters UPLC BEH C18 column by gradient elution using A(methanol/acetonitrile = 1/2) and B(1% formic acid aqueous solution) as the mobile phase at a flow rate of 0.2 ml/min,with the detection wavelength of 268 nm.Results The detection limits were 0.3-0.5 ng,the calibration curves were linear in the range of 2-100 ng(1-50 mg/L),the precision was not more than 4.8% and the rate of recovery were 82.5%-105.6%.Conclusion The method established in the present paper is rapid,sensitive and suitable for the determination of 7 kinds of antibiotics in anti-acne cosmetics.

A method based on HPLC-ICP-MS was established to separate the reaction products of ( ethylenediamine) palladium(Ⅱ) chloride([Pd ( en ) Cl2])and 5’-deoxyguanylic acid ( 5’-dGMP). Two reaction products were detected at pH 8. 0 with 25 mmol/L phosphate buffer solution as chromatography eluent. One was the main product with HPLC retention time of 2. 8 min, the other product’s retention time was 3.2 min. According to ESI-MS(MS/MS) study, m/z=510, 511, 512, 514, 516[M+1]+ parent ions ( abundances same to palladium isotopes) were detected. Further analysis showed that the main product was[Pd( en) ( N1-5’-dGMP) ]. However the other product was hardly to be detected by ESI-MS. By using HPLC-DAD and HPLC-ICP-MS, we found that the two reaction products had the same UV absorption spectra and palladium percentage content. Combined with other groups’research, the other reaction product was deduced as dimmer, trimer or tetramer form of[Pd( en) ( N1-5’-dGMP) ]. Further study revealed that[Pd( en) ( N1-5’-dGMP) ] was easily formed in acid solution while its polymer form was generated in alkaline solution. At pH 6. 0, [Pd(en)(N1-5’-dGMP)] was formed within 12 hours with good stability. Research also revealed that the total amount of two reaction products declined as reaction pH climbed.

A novel method was developed for the determination of the 15+1 European priority polycyclic aromatic hydrocarbons in edible oil by online solid-phase extraction coupled with high performance liquid chromatography-ultraviolet/ fluorescence detection ( online-SPE-HPLC-UV/FL-D ) . The edible oil samples were diluted with isopropyl alcohol, and then filtered. The online extraction was performed on a solid phase extraction ChromSpher Pi column (80 mmí3 mm) and the separation was carried out on a C18 reversed-phase PAH column (250 mmí4. 6 mm i. d, 5μm) using ultraviolet detection at 220 nm and fluorescence detection. Isopropyl alcohol, acetonitrile and water were served as mobile phase in gradient elution. The results showed good linearity for the 15+1 polycyclic aromatic hydrocarbons with all the correlation coefficients (R2)>0. 99. The limits of detection ( LODs ) were between 0. 03 and 12. 23 μg/kg. The recoveries of the sixteen components in the three levels of spiked samples were in the range of 65 . 3%-110 . 5% with the relative standard deviation (RSD, n=6) from 0. 1% to 9. 8%.

Objective To evaluate the efficacy of intrathecally administered monosialoganglioside (GM-1)for treatment of bupivacaine spinal anesthesia-induced neurotoxicity to rat spinal cord.Methods Adult male Sprague-Dawley rats,weighing 280-300 g,in which the intrathecal catheter was successfully inserted into the L3,4 intervertebral space and advanced toward the tail,were randomly divided into 4 groups (n =36 each):sham operation group (group S),GM-1 group,bupivacaine group (group B) and bupivacaine + GM-1 group (group BG).In B and BG groups,the rats received 5％ bupivacaine 20 μl via the intrathecal catheter 3 times at 1.5-hour intervals.GM-1 20 μg was injected intrathecally 24 h later once a day for 7 days in BG and GM-1 groups.Before bupivacaine injection and on days 1,3,5,7,14 and 28 after bupivacaine injection (T0-T6),tail flick latency (TFL) was measured,MPE (percentage of maximal possible effect) was calculated,and the locomotor recovery was evaluated using the Basso,Beattie,Bresnahan (BBB) Locomotor Rating Scale.Then six rats were randomly chosen and sacrificed in each group.Spinal cord was removed for histopathologic examination (with light and electronic microscope) and for determination of caspase-3 protein and mRNA expression (by immuno-histochemistry and RTPCR).The pathological changes of the spinal cord were scored.Results Compared with S and GM-1 groups,MPE,pathological scores,and caspase-3 protein and mRNA expression were significantly increased and BBB score was decreased at T1-6 in group B (P ＜ 0.05),and MPE was increased at T1-5 (P ＜ 0.05) and returned to the baseline value at T6 (P ＞ 0.05) and pathological scores,and caspase-3 protein and mRNA expression were significantly increased and BBB score was decreased at T1-6 in group BG (P ＜ 0.05).There were no significant differences in each parameter at each time point between S and GM-1 groups (P ＞ 0.05).Compared with group B,MPE and caspase-3 protein and mRNA expression were significantly decreased at T2-6,pathological scores were decreased at T3-6,and BBB score was increased at T4-6 in group BG (P ＜ 0.05).The pathological changes of spinal cord tissues were obvious at T1-6 in group B and at T1-3 in group BG,but the changes gradually recovered at T4-6 in group BG.Conclusion Intrathecally administered GM-1 has therapeutic effect against bupivacaine spinal anesthesia-induced neurotoxicity to rat spinal cord,and inhibition of neuronal apoptosis in the spinal cord may be involved in the underlying mechanism.

Objective To investigate the therapeutic effects of intravenous monosialo ganglio-sides(GM-1)on neurotoxicity of intrathecally administered bupivacaine in rats and its possible mecha-nism.Methods One hundred and eight adult male Sprague-Dawley rats,weighing 280-300 g,were randomly divided into 3 groups (n=36 each):sham operation group (group sham),group saline and group GM-1.Neurotoxicity model was performed by injecting 0.12μl/g body weight of bupivacaine at concentrations of 5% via an implanted intrathecal catheter at 90-minute intervals for 4.5 h in groups saline and GM-1.After observing 24 h,group GM-1 was administered GM-1 30 mg/kg by intrave-nous injection for 7 days,once a day;while groups saline and sham received equal volume of normal saline.The recovery of the locomotor function was evaluated with Basso,Beattie and Bresnahan (BBB)and tail-flick latency(TFL)before injection bupivacaine and days 1,3,5,7,14,28 after in-jection,TFL was converted to the percent maximum possible effect (%MPE).Six rats were sacri-ficed in each group at each time point,and spinal cord was taken to examine histological injury scores by light and electron microscopy at the L3 level,and neuron caspase-3 expression was evluated using immunohistochemistry and RT-PCR.Results Compared with group saline,%MPE,histological inju-ry score and caspase-3 mRNA expression were decreased on days 7,14 and 28;Caspase-3 protein ex-pression was decreased on days 5,7,14 and 28;while BBB score was higher on days 14 and 28 in group GM-1 (P < 0.05 ).Compared with group sham,% MPE,histological injury score,caspase-3 mRNA and protein expression in groups GM-1 and saline were significantly higher,while BBB score was lower on 1,3,5,7,14 and 28 d after injection (P <0.05).Conclusion GM-1 can promote neuro-functional recovery after bupivacaine neurotoxicity in rats through the possible mechanism of down-regulating neuron caspase-3 expression.

Purpose:To investigate the value of susceptibility-weighted imaging(SWI)in evaluating the histopathologic grade of cerebral astrocytomas.Materials and Methods: 18 patients with histologically proven cerebral astrocytomas,including 7 diffuse astrocytomas,3 ana-plastic astrocytomas and 8 glioblastomas before treatment were involved in this study.The features on SWI were analyzed in 18 cerebral astrocytomas.Results: The veins in the tumors were not detected in 7 diffuse astrocytomas.Slight edema round tumors appeared in all the diffuse astrocytomas.Plenty veins in the tumors and severe edema round the tumors appeared in 3 anaplastic astrocytomas and 8 glioblastomas.The hemorrhagic foci were detected in 1 anaplastic astrocytomas and 6 glioblastomas.The edema round the tumors were moderate or severe in all the anaplastic astrocytomas or glioblastomas.Conclusions: Susceptibility-weighted imaging can provide the informations about blood supply,hemorrhagic focus and edema round the tumors.SWI is very useful for preoperative evaluation of the histopathologic grade of cerebral astrocytomas,especially for evaluation of high or low grade astrocytomas.

Objective To investigate the effect of vitrification on the biomechanics of tendon tissue in rabbit.Methods Two frozen methods were adopted.The first group was treated with Cryoprotective Agent,which was composed of 18.64%DMSO(V/V),13.37% Acetamide(V/V),9.17% 1,2 Propylene glyco(V/V),0.10mmol/LTrehalose and 10% Calf serum.The tendon tissue with three steps of preliminary treatment,preserved in Cryoprotective Agen was conserved in liquid nitrogen(-196℃)for 14 days;The second group treated with 15% DMSO and 10% Calf serum served as control group.Transmission electron microscope was used to observe the Histiooytic shape of tendon.Each group was performed with tendon tensile test,which could detect maximum load,the maximum shifting quantity and Young's Elastic Modulus.Results There was no significant damage in the tissue's micromechanism of vitrified tendon.But in cryopreservation group,the tissue's micromechanism was apparently damaged.There was no significant difference between test group and control group in maximum load(P=0.256).The same was the maximum shifting quantity(P=0.065).There was significant difference between test group and control group in Young's Elastic Modulus(P=0.006).Conclusion The damage of vitrification to tendon is less than that of profound hypothermia preservion,especially to tendon's corpuscular shape,but there is no significant difference between test group and control group in tendon's biomechanics.

Aim To investigate the protective effect of non-mitogenic human acidic fibroblast growth factor (nm-haFGF) on cerebral ischemia-reperfusion injury in mice. Methods Cerebral ischemia-reperfusion model was made by ligating bilateral carotid for 20 minutes in mice. These mice were randomly divided into model group( iv NS), two doses of nm-haFGF (iv 25、50 ?g?kg-1) groups, rhaFGF group(iv 50 ?g?kg-1) and sham- operated group. Step down test and Y-type electric maze were used to examine the effect of nm-haFGF on learning and memory of mice, then Even′s Blue(EB) level and NO level in brain of these mice were measured. Results The nm-haFGF significantly decreased numbers of errors of mice in 5 min in step down test and in Y-type electric maze test; EB and NO levels in brain of these mice were lower than those of model group respectively. Conclusion The nm-haFGF can protect cerebral ischemia-reperfusion injury in mice.

AstudyofPt-metabolitesfromanewanti-hepatomadrugmiriplatinwasimportanttomiriplatin's pharmacology research. Therefore, a method based on size exclusion chromatography-inductively coupled plasma mass spectrometry ( SEC-ICP-MS) was developed to study miriplatin and its Pt-metabolites in Beagle dog plasma. This method could be used to study total platinum concentration half-quantitatively. Compared with traditional ICP-MS direct determination, data acquired from this SEC-ICP-MS method were almost the same. By using BioSep-s2000 column, 25 mmol/L of pH 7. 2 phosphate buffer as eluent, and Pt-195 as detecting isotope, we discovered miriplatin with its four Pt-metabolites in dog plasma after intra-hepatic artery administration. The main Pt-metabolite was m2 , which associated with plasma proteins. Miriplatin in plasma did not bind with plasma proteins. According to calculation, the ratio of miriplatin/m2 first decreased rapidly, and then slowly increased to its second climax, finally slowly decreased.

Aim: To develop a practical synthetic route of raltegravir, a drug for HIV treatment. Methods: Raltegravir was synthesized through an eight-step process including aminonitrile formation, protection with benzyloxy-carbonyl group, conversion of the nitrile to the amidoxime, cyclization to form hydroxypyrimidinone, N-methyla-tion, amidation with microwave-assistance, deprotection, amidation with acyl chloride. Results: The overall yield of the eight-step synthesis is about 12. 0% and the structure of the target compound was confirmed by ~1H NMR, ~(13)C NMR, LR-MS and HR-MS. Conclusion: The reported synthetic process of raltegravir highlights the advantages in terms of readily available starting materials, convenient operation and low cost.