The course of pharmaceutical analysis for pharmacy undergraduate students in National University of Singapore and China Pharmaceutical University was compared, in terms of curriculum, lecturing, assessment and practical teaching.This study provides in-depth analysis by the lecturers from both universities,and can provide reference for teaching reform and course construction of pharmaceutical analysis.

AIM: To investigate the anti-HIV-1 activity of five anthraquinone derivatives (emodin,rhein,chrysophanol,physcion and aloe-emodin) in vitro.METHODS: Viral replication was estimated by observation of cytopathogenesis and measurement of HIV-1 p24 antigen production in HIV-1ⅢB acutely infected C8166 cells. The anti-HIV-1 activity was evaluated by the 50% effective concentrations (EC50) and selective indexes (SI) of these derivatives.RESULTS: These anthraquinone derivatives inhibited HIV-1ⅢB replication on syncytia formation induced by HIV-1ⅢB infection with EC50 mean values of (11.44±0.93)μmol/L (emodin),(51.28±2.86)μmol/L (rhein),(90.58±2.30)μmol/L (chrysophanol),(8.59±0.38)μmol/L (physcion) and (0.89±0.08)μmol/L (aloe-emodin),respectively. The p24 antigen production with EC50 mean values were (11.61±0.56)μmol/L (emodin),(12.35±4.73)μmol/L (rhein),(39.63±2.87)μmol/L (chrysophanol),>250 μmol/L (physcion) and (2.75±0.20)μmol/L (aloe-emodin) respectively. CONCLUSION: These structurally-related chemicals show different anti-HIV-1 activity in vitro. Among them,aloe-emodin is the most potent inhibitor to HIV-1 replication.

AIM: To investigate the effects of uric acid sodium salt (UANa) as adjuvant on humoral and cellular immune response in BALB/c mice. METHODS: BALB/c mice were immunized with trichosanthin (TCS) as antigen together with UANa suspension as adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay. Dendritic cells (DC) were induced in vitro, the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated. A delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation of lymphocytes was determined by ConA stimulation. RESULTS: Freunds adjuvant greatly enhanced the antibody response of mice to TCS, while UANa adjuvant failed to promote the antibody response but significantly reduced the antibody response as compared to TCS only. No effect of UANa on the expression of CD11c and CD83 in DC was observed by flow cytometry analysis. However, UANa significantly enhanced the expression of MHC II molecule. In the DTH model, UANa enhanced the degree of allergen-induced ear swelling and promoted the ability of lymphocyte proliferation in vitro. CONCLUSION: UANa suspension as adjuvant significantly enhances the cellular immune response but inhibits the humoral immune response to a certain degree, suggesting that UANa has potential application in the vaccine research.

Aim:To develop a practical chiral CE method for the quantitative determination of the unwanted enantiomer[( R )-enantiomer]presented in zolmitriptan. Methods:The background electrolyte was 20 mmol/L sodium dihydrogenphosphate solution with 1% S-β-CD,adjusted to pH 3.50 with phosphoric acid. A fused-silica capillary(60 cm×50 μm ID,effective length 51.5 cm)was used at 20 ℃ for the separation. The applied voltage was -30 kV. The samples were loaded by hydrodynamic injection(50 mbar pressure,6 s). UV was measured at 220 nm. Results:Zolmitriptan and its chiral impurity were baseline resolved ( R s=6.66). The linearity was good over the concentration range from 4 to 80 μg/mL( r =0.999 8) of ( R )-enantiomer. The injection precision (expressed as CV%) was 2.83%. The average recovery was 99.97%( n =9). The limit of detection was 1.5 μg/mL. The host-guest complex binding constants were 964 and 905 mol-1 for ( R )-enantiomer and zolmitriptan,respectively. Conclusion:The method is suitable for the determination of ( R )-enantiomer in zolmitriptan and binding constants of zolmitriptan enantiomers to S-β-CD.

AIM:To investigate the effects of uric acid sodium salt (UANa) as adjuvant on humoral and cellular immune response in BALB/c mice. METHODS:BALB/c mice were immunized with trichosanthin (TCS) as antigen together with UANa suspension as adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay. Dendritic cells (DC) were induced in vitro,the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated. A delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation of lymphocytes was determined by ConA stimulation. RESULTS:Freund's adjuvant greatly enhanced the antibody response of mice to TCS,while UANa adjuvant failed to promote the antibody response but significantly reduced the antibody response as compared to TCS only. No effect of UANa on the expression of CD11c and CD83 in DC was observed by flow cytometry analysis. However,UANa significantly enhanced the expression of MHC II molecule. In the DTH model,UANa enhanced the degree of allergen-induced ear swelling and promoted the ability of lymphocyte proliferation in vitro. CONCLUSION:UANa suspension as adjuvant significantly enhances the cellular immune response but inhibits the humoral immune response to a certain degree,suggesting that UANa has potential application in the vaccine research.

Objective To evaluate the serum 25?hydroxy vitamin D3 level in patients with multiple sclerosis(MS)and normal healthy population,as well as the correlation between addition of oral 1,25?hydroxy vitamin D3 and the prevention of MS relapse and progression. Methods There were 60 cases in the relapsing?remitting MS(RRMS)group and 68 cases in the healthy group,respectively;and the differences in the sex,age, serum 25?hydroxy vitamin D3 level of the two groups were counted and evaluated. In addition ,the 60 cases of patients were divided randomly into the hormone therapy group and the addition treatment group ,with 30 cases in each group;the addition treatment group was added oral calcitriol soft capsules on the basis of the hormone therapy group;EDSS score evaluation was conducted on the two groups 6,12 and 24 months after treatment,the relapse frequency was counted after 24 months,and the relapse interval was calculated. Results The serum 25?hydroxy vitamin D3 levels in the patient group and the healthy group were(18.75 ± 8.35)nmol/L and(23.28 ± 9.31)nmol/L, respectively. There were statistically significant differences in the relapse frequency (P < 0.01),the relapse interval(P < 0.05),and EDSS score(after 24 months)(P < 0.05)between the hormone therapy group and the addition treatment group after treatment;while the differences in the EDSS score (after 6 months)(P = 0.457) and the EDSS score(after 12 months)(P = 0.118)between the two groups showed no statistically significance. Conclusion The serum 25?hydroxy vitamin D3 level in MS patients was markedly lower than that in normal healthy population. Addition of 1,25?hydroxy vitamin D3 contributes to preventing the relapse rate of MS and extending the relapse interval;in addition,maintaining long?term of oral 1,25?hydroxy vitamin D3 facilitates to delaying the progression of disabled disease.

Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.

Objective To discuss the impacts of stroke center construction upon therapeutic indexes for di-agnosing and treating patients with acute ischemic stroke. Methods Patients were divided into a control group (180 patients)and a formal group(245 patients)based on the time of stroke center construction.Patients in both groups were recorded time points in the course of diagnosis and treatment,and compared in number of cases with intravenous thrombolysis,number of cases receiving intravascular interventional therapies,DTN(door-to-needle) time,number of deaths,National Institutes of Health Stroke Scale(NIHSS)scores upon grouping,NIHSS scores after four weeks,NIHSS scores after 3 months,days of hospital stay and hospital charges.Results After the con-struction of the stroke center,time spent in different links were decreased.The number of cases with pure intrave-nous thrombolysis and DTN time(shorter than 60 min)were increased,and the difference were statistically signifi-cant(P<0.01).The number of cases who only received intravascular interventional therapies is increased,mean-while,DTN time was decreased,NIHSS scores after 4 weeks was increased and NIHSS scores after 3 months were also increased(P < 0.05)in these cases. No statistically significant differences existed in number of cases who were treated by bridging with intravenous thrombolysis in combination with intravascular interventional therapies and death cases(P=0.153,P=0.247).There were no statistically significant differences in days of hospital stay and hospital charges(P=0.152,P=0.406).Conclusions After the stroke center construction,it is helpful for significantly improving medical institutions′diagnosis and treatment of stroke,reducing time of such diagnosis and treatment in different links,shortening DTN time,increasing thrombolysis rate,improving patients′prognosis and bringing more benefits to patients with acute ischemic stroke by optimizing procedures for diagnosing and treating stroke.

AIM: To investigate the guiding role of individualized medication adjustment based on CYP2C19 metabolic typing in the treatment of ischemic stroke with clopidogrel, and to provide reference for clinical individualized medication. METHODS: The total of 80 patients with ischemic stroke were divided into the individualized drug instruction group with gene detection (n=40) and the control group without gene detection (n=40) according to whether they received CYP2C19 gene detection. According to the metabolism of CYP2C19, the individualized medication instruction group was divided into slow metabolic type, intermediate metabolic type, fast metabolic type and ultra-fast metabolic type. Patients with fast and ultra-fast metabolites were given clopidogrel dose of 75 mg once a day. Patients with intermediate metabolic type were given double clopidogrel dose of 150 mg once a day. Patients with slow metabolism were given tigrillo dose of 90 mg twice a day or aspirin dose of 100 mg once a day. The control group received 75 mg clopidogrel once a day. All patients enrolled in the groups were followed up for 3 months by outpatients or telephone. The incidence of vascular events and mRS scale scores were compared between the two groups. RESULTS: The incidence of vascular events in the individualized drug instruction group was significantly lower than that in the control group, and the incidence of mRS score(0-1) was significantly higher than that in the control group, with statistically significant differences (P<0.05). CONCLUSION: The individualized medication for patients with ischemic stroke by CYP2C19 gene detection can significantly reduce the incidence of adverse vascular events and improve the prognosis and living ability of patients.

Objective To develop measurement methodology using film for the positioning accuracy of MLC leaves in IMRT.Methods The solid water phantom of 30 cm x 30 cm was scanned and the scanned images were transferred to TPS for treatment plan formulation.The five MLC strip picket fence pattern was formed by MLC leaves,each 3.0 cm long × 6.0 mm wide.The separation between strip and strip is 3.0 cm.SAD is 100 cm at dmax for 6 MV X-ray,with 250 MU per MLC strip.EBT2 radiochomic film was put on the phantom for delivery of IMRT,for each MLC strip.The present study focused on 30 accelerators of Varian,Elekta and Siemens designs at 27 hospitals with highly skilled physicists all over Jiangsu,Sichuan,Hubei and Henan provinces.The study was conducted in the same way as used in international multi-radiotherapy center (IMRC).The irradiated films were sent respectively to IAEA dosimetry laboratory and external audit group (EGA) of China for measurement,analysis and calculation.Results According to IAEA requirements,the differenc of film-measured and TPS-planned of MLC leaf position for each strip should be within ±0.5 mm.China had participated in the research of IMRC,with the result of 0.3,0.2,0.0,-0.1,and-0.2 mm,respectively.For 30 accelerators in four provinces involved in the study,the IAEA's verification results of MLC leaf position were within 0.6-1.0 mm for 5 accelerators and within ± 0.5 mm for other 25 ones.Whereas the verification results of EAG were within 0.6-1.0 mm for 6 accelerators and within ±0.5 mm for other 24 ones.According to IAEA requirements,the film-measured MLC leaf position deviation for each pair of leaves and average all pairs of leaves should be within ±0.5 mm.China had participated in IMRC's research,with the measured result being 0.04 mm.The verification result of EGA for 30 accelerators showed the measured MLC leaf position deviations were all ＜0.3 mm per strip,consistent with IAEA requirements.The IAEA's result showed the measured deviations of MLC leaf position for 29 accelerators were within ±0.5 mm,with only other one being-0.7 mm not consistent with the IAEA requirements.As required by IAEA,the difference of film-measured difference of MLC opening width should be within ±0.75 mm between each pair and average all pairs of leaves.China's result in research of IMRC showed the difference of minimum width to mean width was-0.2 mm whereas the difference of maximum width to mean width was 0.4 mm.For 30 accelerators involved in IAEA'verification study,the measured result shown that the difference between maximum and average of filmmeasured of MLC leaf width,and between minmum and average,were within ± 0.75 mm for 24 accelerators,in line with the IAEA requirements.For other 6 ones,the values were beyond ±0.75 mm,not in line with the IAEA requirements.For the verification result of EAG,the difference between maximum and average widths and between minmum and average widths for 25 accelerators were within ±0.75 mm as required by IAEA,whereas for other 5 ones the value were beyond ±0.75 mm,not consistent with IAEA requirements.The standard deviation of film-measured MLC opening width between each pair and average all pairs should be within 0.3 mm as required by IAEA.China's IMRC result was 0.12 mm.The verification result of IAEA shown the standard deviation of MLC opening position were ＜0.3 mm for 26 accelerators and ＞ 0.3 mm for other 4 accelerators.EAG verification result were the same as IAEA result.Conclusions The method using radiochromic film for measuring accuracy of MLC leaf position is convenient and practicable as a quality audit.It is suitable for quality verification in medical institutions owing to easy to post and repeated measurements.