Aim To establish a cell model for high throughput screening and search for novel inhibitors of P-gp.Methods A fluorescent cell-based assay was developed for application in high throughput screening.A library of 480 traditional Chinese medicine samples was screened. Results The value of Z′ was 0.72. Screening of a library of 480 traditional Chinese medicine identified 2 positive samples.Conclusions This HTS model for screening P-gp inhibitors was stable and sensitive.The 2 positive samples had good P-gp inhibitory activities.

The experimental teaching of Pharmaceutical Technology plays a critical role in improving the students' knowledge conversion,application,and innovation abilities in pharmaceutical engineering specialty.According to the problems existing in experimental teaching process,exploration and practice on the case teaching method and the student-centered teaching mode are introduced in this paper.The implementation of the teaching reform can effectively combine pharmaceutical technology theory with practice,and is conducive to the professional development of pharmaceutical engineering in our university.

Objective To study the molecular genetic mechanism of a patient with 17? hydroxylase (CYP17) deficiency. Methods Genomic DNA were abstracted from the blood of the patient, her parents and healthy control. The 8 exons of CYP17 gene were amplified, using 5 pairs of designed primers, with polymerase chain reaction (PCR), and the 8 exons were sequenced by the dideoxy terminator method to determined the mutation sites. The corresponding exons of the parents of the patients were also amplified and sequenced to determine the zygosity of the patient and the source of the gene variances. Results The analysis revealed that the patient (46, XY) was a compound heterozygote carrying two different inherited mutations on CYP17 gene, one from mother containing a point mutation Arg 96 (C G G)→ Gln(C A G) and the other from father containing a nine base deletion (CACTCTTTC) at amino acid position 487～489 (Asp Ser Phe) near the carboxyl terminus of P450c17. Conclusion The CYP17 gene of the patient with 17? hydroxylase deficiency is a compound heterozygous mutation. The mutation changes the amino acid sequence of P450c17 enzyme, which in turn affected the enzymatic activity. Arg 96 is essential in P450c17 enzyme activity. Deletion of Asp 487 Ser 488 Phe 489 in exon 8 may be a prevalent mutation causing P450c17 deficiency in Southeast Asia.

Objective To investigate the changes of calmodulin kinase Ⅱ (CaMK Ⅱ) and cAMP responsive element binding protein(CREB) expressing in primary cultured hippocampal neurons and its relationship with learning and memory deficit after 2000 μW/cm2 electromagnetic radiation.Methods Primary cultured hippocampal neurons in vitro were randomly divided into normal control group,sham-radiated group,and 1 h/d,2 h/d,3 h/dradiation groups.The neurons in the radiation groups were received microwave exposure of 2000 μW/cm2.The change of CaMK Ⅱ and CREB protein in hippocampal neurons of each group of rats were measured with western blot,and the expression of CaMK Ⅱ and CREB mRNA in hippocampus were determined by RT-PCR.Results Compared with control group((0.78 ± 0.07),(0.62 ± 0.12)),the expression of CaMK Ⅱ protein (1 h/d(0.59 ±0.05),2h/d(0.44 ±0.08),3h/d(0.18 ±0.04)) and its mRNA(1h/d(0.41 ±0.08),2h/d(0.34 ±0.04),3h/d(0.24 ±0.02)) was obviously decreased (P＜0.05).Compared with control group((0.69 ±0.10),(0.80 ±0.12)),the expression of CREB protein(1h/d(0.49 ±0.05),2h/d(0.4 ±0.04),3h/d(0.17 ±0.03))and its mRNA (1 h/d (0.68 ± 0.11),2h/d (0.53 ± 0.08),3h/d (0.30 ± 0.03)) was obviously decreased after radiation(P＜0.05).Conclusion Electromagnetic radiation of 2000 μW/cm2 exposure could weaken the learning and memory abilities of rats and the decreases in the expression of CaMK Ⅱ and CREB protein and their mRNA in hippocampus may be involved in the pathophysiological process of learning and memory deficit.

Objective To determine the diagnostic value of SPECT in lower digestive tract hemorrhage (LDTH). Methods Of the 24 LDTH cases,16 were tested by gastric mucosa imaging ,4 by 99m Tc PHA imaging,and the other 4 by 99m Tc RBC imaging.All the patients' diagnosis were confirmed by surgery and pathology. Results The sensitivity and the specificity of the gastric mucosa imaging were 100%(16/16) and 93.7%(15/16),respectively. The sensitivity of the 99m Tc PHA imaging and the 99m Tc RBC imaging was 75.0%(3/4) and 75.0 %(3/4), respectively, but had no specificity. Conclusions SPECT has very high sensitivity and specificity in the diagnosis of Meckel's diverticulum. 99m Tc PHA imaging and 99m Tc RBC imaging may help to determine the position of bleeding in unkown causes of LDTH.

Objective To analyze the clinical effect of vacuum sealing drainage (VSD) on skin and soft tissue injury in elderly patients.Methods A total of 75 patients with severe skin and soft tissue injury who met the inclusion criteria were enrolled in this study.They were divided into two groups based on admission dates:the observation group (n =40,receiving VSD treatment,oddnumber date admission) and the control group (n=35,receiving routine dressing changes,even number date admission).The clinical indexes including pain score,wound healing time,infection rate and complications were analyzed and compared between the two groups.Results There was a significant difference in the average length of hospitalization between the two groups (21.3 days vl 30.7 days,t=7.60,P=0.0000).The infection rate was lower in the observation group than in the control group (12.5％ or 5/40 vs.54.3％ or 19/35,x2=13.12,P=0.0003).The incidence of complications was lower in the observation group than in the control group (15.0％ or 6/40 vs.42.9％ or 15/35,x2=5.87,P=0.0154).There were no deaths in the observation group,but one patient died from acute pulmonary embolism in the control group during hospitalization.Conclusions VSD can alleviate pain,reduce the infection rate,shorten the length of hospitalization and decrease bedridden complications in elderly patients with severe skin and soft tissue injury,and has valuable applications in clinical practice.

AIM: Construction of an eukaryote- E. coli shuttle expressing recombinant plasmid which expresses OmpL1 envelope protein of pathogenic Leptospira, serovar Lai strain 017. METHODS: The OmpL 1 gene was amplified by PCR from the leptospiral genome. Then it was cut with restriction enzymes and ligated to the plasmid pBK-CMV. The correct recombinant plasmid was screened out with analysis of restriction enzymes and PCR. After inducing the E. coli baring recombinant plasmid with IPTG,the complete protein of the bacteria was extracted for SDS-PAGE. At the same time, OD600 of the host bacteria was examined at different time after inducing or uninducing with IPTG. RESULTS: Five strains E. coli containing proper recombinant plasmids were screened out. Four strains E. coli expressed a new protein with a weight of 37 kD among them. With the expression of the heterogenous protein,the OD600 of the host bacteria decreased. CONCLUSION: The shuttle expressing plasmid of the OmpL 1 gene of strong virulent Leptospira strain 017 was successfully constructed. Furthermore,the recombinant plasmid expressed the expected OmpL 1 fusion protein in E. coli and the expression of the heterogenous protein had toxic effect on the host bacteria. This work was important for the future research of OmpL1 protein which relates to the diagnosis,new vaccine preparing and the pathogenic mechanism of leptospirosis.

Objective To probe into the feasibility and efficacy of damage control surgery (DCS) in treating severe pelvic fracture combined with abdominal organ injuries.Methods A retrospective analysis was done on the clinical data of 39 patients with severe pelvic fractures combined with abdominal organ injuries treated by DCS from 1995 to 2010.Results Devascularization of internal iliac arteries was performed to treat massive hemorrhage in the 39 patients with severe pelvic fractures combined with abdominal organ injuries,including ligation of bilateral internal iliac arteries in 31 palients and angioembolization of bilateral internal iliac arteries in eight.Meanwhile,early pelvic external fixators were used in 31 patients.All patients received discriminating internal fixation after resuscitation in ICU.The overall mortality rate was 21％ (8/39),with average ISS of 41.6 points and with hemorrhagic shock and combined injury for the main causes of death.Complications occurred in seven patients including combined acute respiratory distress syndrome (ARDS) in three patients,thrombosis of right common iliac artery in one,subphernic abscesses in two,and deep infection of lower extremity in one,but all the complications were cured.Conclusions Reasonable and timely use of DCS can enhance the rescue survival rate of patients with severe pelvic fraclure combined with abdominal organ injuries.

Objective To discuss the flail chest in the rib the fixed surgery opportunity,in fixed spot. Meth-ads Clinical data of 26 flail cheat patients in my courtyard were retrospectively analyzed. Results 26 patients were cured completely,nobody died,no obvious cheat abnormity was observed. Condusion Regarding the flail cheat pa-tient, at the right moment to adopt the fixed surgery in the rib, restore the integrity of the thoracic wall, may reduce the mortality rate, the chest shape normal, the quality of life obtains the enhancement.