OBJECTIVE: To develop in 6-year grade postgraduates the expert knowledge and capability in clinical pharmacy through systematic training and practice. METHODS: In the early post-graduate stage,the postgraduate students were assigned to different specific areas. Training methods such as clinical tutoring system,multi-angle training,regular training etc were taken,and a training procedure of pharmacy practice,clinical practice and joining in specialized subjects etc was set up for the students. RESUTLS: After training,the students had adapted to the routine work in hospital pharmacy,developed the knowledge and skills to provide clinical pharmaceutical care,and mastered the clinical pharmacy research methodologies to set up subjects and solve clinical problems. CONCLUSION: The training program for the 6-year grade postgraduates majored in clinical pharmacy achieved a satisfactory outcome in either practice or scientific research.

OBJECTIVE:To develop a simple micellar electrokinetic capillary chromatography for determination of theophylline concentration in human plasma. METHODS: The determination of plasma sample(without any pretreatment) was performed on un-coated fused-silica capillaries (at an effective length of 21 cm) in which the cathodic buffer solution (solution A) was 15 mmol?L-1 borax(pH=10) and the running buffer solution(solution B) was sodium dodecyl sufate (SDS)(200 mmol?L-1) -contained solution A. The sample was injected into capillary by pressure with separation voltage of 12 kV;the UV detection wavelength was set at 280 nm;quantitation was performed by external standard peak area method. RESULTS: The average determination time of each sample was 11 min;the linear range of theophylline was 1.25～40 ?g?mL-1 with its lowest detectable limit at 0.5 ?g?mL-1;the average methodological recovery was 90.33%(RSD=2.51%);the intra-day RSD was 2.34%～3.36% and the inter-day RSD was 2.03%～6.51%,respectively. CONCLUSION: The developed method is simple,rapid and sensitive and it is applicable for the therapeutic drug monitoring of theophylline.

OBJECTIVE: To investigate the expression of CXCR4 in maxillary sinus carcinoma cells IMC3 under hypoxia. METHOD: IMC3 cells were cultured for 6 h, 12 h, 24 h and 48 h under normoxia and hypoxia. Real-Time PCR was applied to detect the expression of mRNA of CXCR4 and immunohistochemisrty was applied to investigate its protein level. RESULT: CXCR4 mRNA level was about 0.035 under normal conditions, which was obviously upregulated by hypoxia. The mRNA levels after culturing under hypoxia for 6 h, 12 h, 24 h and 48 h were 0.283, 0.313, 0.426, 0.510 respectively. There was statistically significant difference between the mRNA levels of each two groups (P < 0.05, Mann-Whiney Test) with a time dependent course, except for the difference between the groups of 6 h and 12 h. Immunohistochemistry showed that there was almost negative staining for CXCR4 in the cell cultured in nomoxia, while stong positive staining of CXCR4 was observed in cells cultured in hypoxia . The positive staining was located mainly in the cell membrane and cytoplasm and little in the nucleus. CONCLUSION Hypoxia could induce expression of CXCR4 in IMC3 cells at both mRNA and ptrotein levels. The upregulation of CXCR4 by hypoxia showed an obvious time dependent course.
Cell Hypoxia ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Maxillary Sinus ; metabolism ; pathology ; Receptors, CXCR4 ; metabolism

Objective　To explore the relationship between hemolymph phenol oxidase and the melanization of Plasmodium yoelii oocysts in Anopheles dirus. Methods　An Anopheles dirus-Plasmodium yoelii system was used Anopheles dirus were divided into 3 groups, that is, non-blood-fedding (N), normal-blood-fedding (B) and infected-blood-fedding (I). The activities of MPO and o-DPO in hemolymph from 3 groups were determined with native polyacrylamide gel electrophoresis (PAGE) and density scanning at 5, 7, 11 and 15 d after blood feeding. Results　Both MPO and o-DPO activity were significantly higher in group I than group N and B (P<0.05). But with the melanization of Plasmodium yoelii oocysts, both MPO and o-DPO activity in group I were decreased in comparison with group N, especially on the 15 th day after infected-blood feeding. MPO and o-DPO activity in group B were significantly stronger than those of group N. Conclusion　Blood feeding and infection of Plasmodium yoelii both can activate the cascade. The heamolymph phenol oxidase may play an important role in the melanization of Plasmodium yoelii oocysts in Anopheles dirus.

Objective To observe the effect of blood feeding and plasmodium yoelii infection on the transcript abundance of ribosomal protein S7 from Anopheles dirus hemocyte Methods Anopheles dirus of 3～5 days old were divided into normal group (N), blood feeding group(B) and Plasmodium yoelii infection group(I) Hemocytes of 50 Anopheles dirus from each group were collected by centrifuge method on days 1, 2, 3, 4, 7 and 11 after blood feeding, respectively Then, all hemocyte samples were used for total RNA isolation and analyzed by RT PCR Finally, the same volume(10 ?l) of all the PCR products from each group was used for agarose gel electrophoresis and the data obtained were analyzed statistically Results There was no significant difference in ribosomal protein S7 signal between the three groups Conclusion Similar to Anopheles gambiae and human rpS7, Anopheles dirus rpS7 might be also used as an internal control for the studies of the role of Anopheles dirus related immune factors in attacking Plasmodium infection

Objective To investigate the relationship between the immune defence reaction against Plasmodium infection and the prophenoloxidase (PPO) of the midgut by comparative analysis of the distributions and the changes of PPO in the midgut of Anopheles stephensi and Anopheles dirus before and after infection with Plasmodium yoelii . Methods Midguts were dissected out from both Anopheles stephensi and Anopheles dirus at 3, 5, 7, 11 and 15 d before and after infection with Plasmodium yoelii . Immunohistochemistry and Western blotting were performed respectively on the collected midguts using Manduca Sexta PPO IgG polyantibody. Results PPO in the midguts from both Anopheles stephensi and Anopheles dirus was mainly located in the circulation conduit of midgut before infection with Plasmodium yoelii , but aggregated and distributed at the interspace of midguts as pieced or/and stripped forms after infection. Furthermore, PPO in the midgut of Anopheles dirus was more concentrated than that of Anopheles stephensi . Western blotting revealed that the PPO band with about molecular weight of 67?10 3 was detected in the midguts of both Anopheles stephensi and Anopheles dirus before and after Plasmodium yoelii infection. There was significant difference before and after infection, and the PPO band was obviously enhanced after infection. PPO bands in the midgut of Anopheles dirus were more prominent than those of Anopheles stephensi . Conclusion PPO in the midgut of Anopheles mosquitoes may come from the hemolymph by the circulation conduit before Plasmodium yoelii infection. However, the different distributions and changes of PPO in the midguts resulted from the Anopheles mosquitoes infected with Plasmodia may be closely correlated with Plasmodia infection, which may be of important physiological significance and may be involved in the immune defensive reaction against Plasmodium .

Objective To clone the partial cDNA sequence of prophenoloxidase (PPO) gene of Anopheles dirus . Methods Degenerative primers were designed according to the conserved sequence blocks within the prophenoloxidase of insects. RNA sequence of the larva of Anopheles dirus was amplified by RT PCR to get the prophenoloxidase cDNA which was then cloned into T vector and sequenced. The partial cDNA sequence of prophenoloxidase gene was analysed and compared with other prophenoloxidase gene of insects. Results The partial cDNA sequence of AdPPO4 was 597 bp, and its deduced amino acid sequence was 199aa. The cDNA sequence homology and amino acid sequence homology was 84% and 90%, respectively, as compared with the PPO4 gene of Anopheles gambiae . Conclusion The AdPPO4, with high sequence homology with the PPO4 gene of Anopheles gambiae , is successfully cloned from the larva of Anopheles dirus .

Objective To study the change of intracellular free Ca2+ in the oocyst when it melanized and to find out the relationship between the melanized oocyst and its intracellular level of free Ca2+ in a Plasmodium-refractory strain of Anopheles dirus. Methods The distribution and experimental condition of the intracellular free Ca2+ in oocyst of Plasmodium yoelii was measured with Ca2+ sensitive dye Fluo-3/AM and Plutonic F-127 under confocal laser scanning microscope (CLSM) at different time. Results The best load condition was that the oocysts were incubated in 3 ?mol/ml Fluo-3/AM adding 1 ?l/ml 25% Pluronic F-127 for 60 min at 37 ℃ . Fluorescent imaging of oocysts was affected by an increase or decrease of the concentration of Fluo-3/Am and incubation time. The distribution of intracellular free Ca2+ was heterogeneous in the oocysts. The mean value of Ca2 + in the mature oocysts was (137.15 ?7.02) nmol/L(X?S) but was (18.44? 1.75) nmol/L in melanized oocysts with Ca2+ sedimentation in the wall of oocyst. Conclusion The results suggest that the level of the intracellular free Ca2+ in oocyst decreased and excreted during its melanization in a Plasmodium-reiractory anopheline mosquito species.

Objective To analyze the relationship between the TEP1 gene of Anopheles stephensi and melanotic encapsulation of Plasmodium yoelii induced by anti-malaria drug nitroquine. Methods Haemolymph samples from three groups of An. stephensi fed with sucrose solution, Plasmodium-infected blood and nitroquine, respectively, were collected at the 1st, 2nd, 3rd and 4th day after drug adminstration. Degenerate primers were designed according to the conserved amino acid sequence within TEPs of the mosquitoes. Fluorescent quantitation PCR was used to detect the variation of TEP1 gene transcript induced by nitroquine. The melanization of oocysts was observed by light microcopy. Results TEP1 gene was cloned, the predicted amino acid residues harbored a highly conserved canonical thioester motif GCGEQ. The fluorescent quantitation PCR revealed that nitroquine induced an up-regulation of TEP1 activity. The transcription of TEP1 gene in nitroquine treated group (2.423) was significantly higher than that of the infected blood-fed group(1.036) at the 3rd day after nitroquine treatment (P