OBJECTIVE:To provide reference for playing pharmaceutical support role of clinical pharmacist to offer medical assistance in the emergence such as earthquake.METHODS:Basing on the experience in Wenchuan Earthquake,the character and forms of earthquake medical assistance were introduced,then the role of clinical pharmacist in earthquake medical assistance were analyzed.RESULTS:Clinical pharmacists distributed effectively to the drug supply so as to save medical resources and ensure rational administration during the medical rescue in earthquake.CONCLUSIONS:For achieving better function of the treatment team,national medical rescue team should involve clinical pharmacist.

ObjectiveTo investigate the effects of Shen-wu capsule on learning and memory ability and its mechanism in APP transgenic mice. MethodsThe APP 695V717I transgenic mice were randomly divided into model group, Shen-wu low dose group (0.4g/kg·d) and high dose group (1.2g/kg·d). Normal control adopted the same age and background C57BL/6J mice. The animals were administered intragastrically by the drug or water from 4 month old to 10 month old. Morris water maze and object recognition test were performed to measure the learning and memory ability. The content of β-amyloid (Aβ) in the brain cortex homogenate was detected with RIA,and amyloid plaques were measured with Congo red staining. ResultsIn the Morris water maze test, swimming time and swimming distance of model group were prolonged distinctly(P<0.01). Shen-wu high dose group obviously shortened swimming time(P<0.05). In the object recognition test, the relative time to the new objection in model group was obviously shorter than that in the control group(P<0.05). The relative time to the new objection for Shen-wu high dose group was obviously longer than the model group(P<0.05). The content of soluble Aβ in model group was higher than that of the control group(P<0.05). Shen-wu group decreased the soluble Aβ distinctly(P<0.01). The amyloid plaques were increased in the brain of model mice(P<0.01). Each group of Shen-wu decreased amyloid plaques significantly(P<0.01).ConclusionShen-wu Capsule ameliorated the learning and memory function disorder and decreased Aβ formation in the brain of APP transgenic model mice.

In our gastroenterology clinic from Jun. 2008 to Jan. 2009, 20. 9% (277/1320) patients were diagnosed with functional dyspepsia (FD) and 14. 8% (41/277) of them with psychological problems according to Minnesota Multiphasic Personality Inventory (MMPI). In those with psychological disorders,92.7% were found with somatization, 60. 9% with depression and anxiety, 97.6% with abdominal pain,75.6% with bloating, 63.4% with early fullness and 36. 6% with nausea. More than 80% patients with anxiety and depression complained irritability, worry, fatigue, poor concentration, memory loss and insomni.a. After 4 weeks of psychological consultation and anti-depression treatment, 5 patients had significant improvement, 31 had improvement and 3 had no response. The overall response rate was 87. 8%.In summary,there is a high prevalence of depression and anxiety in population with FD. Hypochondria and somatization are common among these patients. Psychological consultation with anxiolytic drugs may have good therapeutic effects.

Objective To explore the effects of 2,3,5,4'-tetrahydroxy stilbene-2-β-D-glycoside(TSG)on the over-expression and aggregation of α-synuclein in vitro.Methods TSG in different concentrations was incubated with α-synuclein transgenic COS-7 cells for 24 h.The cell viability was measured by MTT method.The expression of α-synuclein protein was determined by immunocytochemistry and Western blotting method.Results Incubation of TSG at the range of 12.5～200.0 μmol/L with α-synuclein transgenic COS-7 cells for 24 h did not influence cell viability,but a dose-dependently inhibition for the over-expression of α-synuclein protein could be observed in the tests of immunocytochemistry and Western blotting.Conclusion TSG can inhibit the over-expression of α-synuclein protein in COS-7 cells in vitro.

Objective To evaluate the influence of duration of laser irradiation on histomorphometric meas- urements in experimentally tenotomized and repaired rabbit Achilles tendons and to explore the best irradiation time. Methods A total of 20 male New Zealand rabbits aged 10-12 weeks were used and randomly divided into 4 groups:a control group and three experimental groups.All the animals underwent surgical excised and then repair of their Achillis tendon.The animals in the control group were then treated with sham laser irradiation,while those in the three experimental groups were treated with 10,20 and 30 minutes of He-Ne laser irradiation(632.8 nm, 18.9 mW)daily,respectively,for 14 days.On the 28th day after surgical operation,the animals were sacrificed and their Achilles tendons were sampled.HE stain and Van Geison stain were used to observe morphometric changes of tendons.The SDS-PAGE electrophoresis method and CS-930 photodensity scan instrument were employed to measure the content of typesⅠandⅢcollagen.Results it was shown that laser irradiation enhanced cell proliferation,cel- lular content,granulation tissue formation and collagen deposition in laser-treated tendons,especially in those irradia- ted for 20 minute daily,as compared to the control group.TypeⅠand typeⅢcollagen levels were significantly in- creased at the 28th day in the healing tendons and the ratio of collagenⅢtoⅠincreased in all the 3 experimental groups,and the increase of both collagen content and ratio of collagen typeⅢtoⅠwas significantly greater in those ir- radiated 20 minutes daily(P

Objective To research the effects of transcutaneous electrical stimulation (TES) on the expression of neurotrophin-3 (NT-3) and tumor necrosis factor-α (TNF-a) in the ventral horn of rats' spinal cords and in the injured region after spinal cord injury (SCI),and to explore the effects of TES on neuron reconstruction and functional recovery and their mechanisms. Methods Forty-eight Wistar rats were selected and divided using stochastic methods into a model group and a TES group.Using Allen's method,a complete SCI model was created at T9.Rats of the TES group were given TES treatment.Basso-Beattie-Brasnahan ( BBB ) ratings were used to evaluate locomotor function.Both groups were sampled at 1,3,5 and 7 days after the operation.Immunohistochemical techniques were used to detect the expression of NT-3 and TNF-α in the rats' spinal cords at the different time points. Results The post-operative BBB ratings of both groups showed an increasing trend.In the TES group the improvement was significantly better at 5 and 7 days than in the model group.The expression of NT-3 immuno-positive cells increased in both groups,peaking at 5 days post-operation,then declining at day 7.The expression of NT-3 positive cells at days 5 and 7 had increased significantly more in the TES group than in the model group.TNF-α immuno-positive expression increased with time in both groups,but in the TES group the expression increased substantially less than in the model group.At days 5 and 7 post-operation,the expression was significantly lower than in the model group. Conclusions TES can promote NT-3 expression in rats with SCI,inhibit the increase in TNF-α expression,and aid repair and reconstruction of neurons and related functional recovery.

BACKGROUND:Human mesenchymal stem cells (hMSCs) become aging and even die after several passages. Some investigations have shown that telomere has a close correlation with life span of the cells. Whether the ectopic expression of human telomerase reverse transcriptase (hTERT) could induce the activity of the telomerase, maintain the length of telomere, and finally prolong the life cycle of MSCs without losing their multipotent differentiation capacity is still uncertain.OBJECTIVE: To observe the influence of the ectopic expression of hTERT on the telomerase activity and cell life cycles of hMSCs.DESIGN: Repetitive measurement trails.SETTING: Research Center of Stem Cell, Zhengzhou University Medical College.MATERIALS: The experiment was conducted in the Research Center of Stem Cell, Zhengzhou University Medical College from October 2003 to December 2005. hMSCs were obtained from 20 healthy donators from the Department of Pediatric Surgery and Outpatient, the Third and First Affiliated Hospitals of Zhengzhou University. Enhanced green fluorescent protein plasmid (pEGFP-C1) and pEGFP-hTERT were provided by Dr. Chantal Autexier of Canada. DH5α strain provided by Dr. Hou Wei-hong, the Key Molecular Medical Laboratory of Zhengzhou University Medical College.METHODS.: Under sterile condition, 2 mL bone marrow of sternum of healthy donors were harvested, and prepared after centrifugalization,dilution and passage.① Transfection of pEGFP-hTERT into hMSCs and the screening and amplification of resistance cloning:The 5th passage cells were seeded in a 24-well plate,and transfected by pEGFP-hTERT with lipofectamine method.The cells were divided into four groups including untransfected group,lipofectamine group,pEGFP-C1 group and pEGFP-hTERT group. Resistance cloning screen and amplification was performed by G418. ②hTERT mRNA expression and detection of telomerase activity:RT-PCR and PCR-ELISA were used to detect the hTERT mRNA expressions of the fifth passage hMSCs transfected with pEGFP-hTERT, and pEGFP-C1, the untransfected tenth passage hMSCs and K562 cells (positive control), and the telomerase activity of the fifth and thirtieth passage hMSCs transfected with pEGFP-hTERT,the fifth pEGFP-C1-transfected cells and the tenth passage untransfected cells. ③Karyotype analysis of hTERT-transfected MSCs: Chromosome analysis was performed by conventional Giemsa staining.④Inducement of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification:The transfected MSCs were cultured in a medium containing epidermal growth factor and basic fibroblast growth factor, which could induce the cells differentiate into neuron-like cells. The culture solution was changed every 3 days, and the changes in cell growth condition and morphologic characteristics were observed under an inverted microscope. The microtubule associate protein (MAP2) and neurofliament subunit M (NF-M) were identified by RT-PCR.MAIN OUTCOME MEASURES:①hMSCs transfection with different kathion liposomes and the screening and amplification of resistance cloning; ②hTERT mRNA expressions of each group and detection of telomerase activity; ③Karyotype analysis of pEGFP-hTERT-transfected MSCs; ④ Induction of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification.RESULTS: ①With the decrease of G418 concentration, the cells in the untransfected and lipofectamine groups died, and stably EGFP expressed MSCs were obtained; after G418 screening, there was a cell clone undergone 35 passages and continued to proliferate, whose appearance and growth characteristics were similar to the untransfected MSCs observed under inverted microscope. ②The fifth passage pEGFP-C1-transfected hMSCs and tenth passage untransfected hMSCs remained telomerase-negative, but the K562 and fifth passage hTERT-transfected cells showed positive telomerase activity. ③The telomerase activity of the fifth and thirtieth passage hTERT-transfected cells was positive. ④The hTERT-MSCs at passage 10, 20 and 30 had 23 pairs of chromosomes, and two X chromosomes. So they were still normal diploid with normal chromosome appearance and number. ⑤Many hTERT-transfected MSCs had the typical appearance of neuron-like cells. RT-PCR analysis showed that th expressions of MAP2 and NF-M were increased.CONCLUSION:Ectopic expression of the hTERT gene is found in hMSCs,and can induce the telomerase activity of hMSCs.The ectopic expression of the hTERT gene in hMSCs could extend the life spans of cells and maintain their multipotent differentiation capacity.

OBJECTIVE:To establish the method for the determination of main components content and entrapment efficiency of transferrin modified vincristine-tetrandrine liposomes. METHODS:HPLC method was adopted to determine the content of vin-cristine. The determination was performed on ELITE C18 column with mobile phase consisted of methanol-15%triethylamine (70∶30)at a flow rate of 1.0 ml/min. The determination wavelength was set at 297 nm and column temperature was 30 ℃. The sample size was 20 μl. Free drug was isolated from liposomes by dextran gel column chromatography,and entrapment efficiency was deter-mined. RESULTS:The linear range of vincristine was 160-1 600 μg/ml (r=0.999 8,n=5) with average recovery of 99.20%(RSD=0.26%,n=9). RSD of precision test was 0.070%(n=5). The average content of vincristine in liposomes was 0.790 mg/ml(RSD=0.15%,n=3),and the average entrapment efficiency was 85.94%(RSD=2.08%,n=3). CONCLUSIONS:The method is accurate,reliable,simple and rapid,and can be used for the determination of the content and entrapment efficiency of vincristine in transferrin modified vincristine-tetrandrine liposomes.

OBJECTIVE:To prepare transferrin(TF)modified tetrandrine(TET)and vincristine(VCR)active targeting lipo-somes,and to optimize its formulation. METHODS:TF modified TET and VCR liposomes were prepared by ammonium sulfate gradient method. Using comprehensive score of encapsulation efficiency of TET and VCR as index,central composite design-re-sponse surface method was used to optimize and validate mole ratio of EPC/Chol,mole ratio of EPC/PEG2000-DSPE and TF mass fraction. RESULTS:The optimal formulation was that the mole ratios of EPC/Chol and EPC/PEG2000-DSPE were 1.5:1 and 20:1, TF mass fraction was 0.10%. The encapsulation efficiency of TET and VCR were 97.80% and 93.00%,respectively. The compre-hensive score was 94.44(n=3)which was close to the predicted value of 93.81. CONCLUSIONS:The optimal formulation is sta-ble and can be used for the preparation of TF modified TET and VCR liposomes.