Objective To investigate the distribution of related single nucleotide polymorphisms (SNPs) of drug transporters in healthy Chinese Han population.Methods 10 specific SNPs of 6 drug transporters,including organic anion transporting polypeptide 1B1 (OATP1B1),OATP1B3,organic cation transporter 1 (OCT1),P-glycoprotein (P-gp),multidrug resistance protein 2 (MRF2) and breast cancer resistance protein (BCRP),were determined in healthy Chinese Han volunteers by the established rapid polymerase chain reaction (PCR) methods and sequencing.Gene frequencies were calculated and compared with other populations in the 1000 genomes project.Results The minor allele frequencies of OATP1B1 388A ＞ G,OATP1B1 521T ＞ C,OATP1B3 699G ＞ A,OCT1 181C ＞ T,OCT1 1393G ＞ A,OCT1 1258delA,MDR1 3435T ＞ C,MRP2-24C ＞ T,BCRP 421C ＞ A and BCRP 376C ＞ T were 26.97％,6.25％,23.91％,0,0,0,38.47％,18.52％,31.97％ and 1.48％,respectively.By comparing the distribution of allele frequencies between test group and Mrican,the frequencies of the OATP1 B3 699G ＞ A variant alleles were 76.09％ and 35.63％,respectively;the frequencies of the MDR1 3435T ＞ C variant alleles were 61.53％ and 85.02％,respectively;the frequencies of the MRP2-24C ＞ T variant alleles were 18.52％ and 3.10％,respectively;the frequencies of the BCRP 421C ＞ A variant alleles were 31.97％ and 1.29％,respectively.And these differences were statistically significant (P ＜ 0.05).By comparing the distribution of allele frequencies between test group and American,the frequencies of the OATP1B1 388A ＞ G variant alleles were 26.97％ and 47.26％,respectively;the frequencies of the BCRP 421C ＞A variant alleles were 31.97％ and 14.12％,respectively.And these differences were statistically significant (P ＜ 0.05).By comparing the distribution of allele frequencies between test group and European,the frequencies of the OATP1B1 388A ＞ G variant alleles were 26.97％and 40.26％,respectively;the frequencies of the OATP1B1 521T ＞ C variant alleles were 6.25％ and 16.10％,respectively;the frequencies of the OCT1 181C ＞ T variant alleles were 0 and 6.26％,respectively;the frequencies of the BCRP 421C ＞ A variant alleles were 31.97％ and 9.44％,respectively.And these differences were statistically significant (P ＜ 0.05).There was no significant difference in the distribution of allele frequencies between test group and Japanese (P ＞ 0.05).Conclusion Significant differences were observed in the distributions of some SNPs between Chinese and other populations.It is important to analyze the distribution of the related genetic polymorphisms of drug transporters in Chinese people to guide rational drug use in clinical.

Objective: To investigate the safety and efficacy of pulmonary vein deployment technique for percutaneous closure of atrial septal defects (ASD) solely under echocardiography guidance. Methods: A total of 38 ASD patients received pulmonary vein deployment in our hospital from 2012-10 to 2016-09 since the conventional method could not deliver the occluder to correct place. The patients were with the mean age at (16.0±15.6) years, body weight at (37.2±22.9) kg and ASD diameter at (17.1±4.2) mm. Operative effect was assessed by echocardiography. Follow-up study was conducted at 1, 3, 6, 12 months post-operation and at each year thereafter. Results: 37 patients were successfully finished pulmonary vein deployment for percutaneous closure of ASD solely under echocardiography guidance. One patient was successfully treated by a controlled steerable sheath. The mean operative time was (25.2±5.1) min and mean diameter of ASD occluder was (22.9±5.6) mm. 2 patients had trivial residual shunt at the early post-operative stage. No peripheral vascular injury, pulmonary vein and cardiac perforation occurred. All 38 patients were recovered and discharged. The average in-hospital time was (2.9±0.7) days. The patients were followed-up for (23.9±15.4) months, without complications of residual shunt, pericardial effusion, aortic regurgitation and pulmonary vein stenosis. Conclusion: Pulmonary vein deployment technique for percutaneous closure of ASD solely under echocardiography guidance was safe and effective; it can avoid radiation damage and provided a simple and practical method for ASD patients who failed to conventional method under echocardiography guidance.

Objective To study the antiviral effect of Jin Qiao Tablets on influenza A H1N1 virus in vivo. Methods The mouse pneumonia model was established by nasal inhalation of 15 LD50 of influenza virus. After prophylactic or therapeutic medication for 5 d,mouse lung tissue was taken out and weighed. Viral load in lung tissue was measured by polymerse chain reaction(PCR),and the level of γ-interferon(γ-IFN)in rat serum and lung was detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA)for evaluating the effect of Jin Qiao Tablets on lung index, viral load and γ-IFN in rats. After prophylactic or therapeutic medication for 7 d,morbidity and mortality within 14 d of mice with pneumonia induced by nasal inhalation of 3 LD50 were observed to evaluate the action of Jin Qiao Tablets for protecting against death and prolonging life span. Results Jin Qiao Tablets markedly decreased the increased lung index,promoted the death-protection rate and life-prolongation rate, decreased viral load, raised the level of γ-IFN in mice (P < 0.05 or P < 0.01). Experimental results in vivo showed that Jin Qiao Tablets had better anti-influenza virus activity than Yinqiao Jiedu Tablets and Lianhua Qingwen Capsules, and the effect of Jin Qiao Tablets was equivalent to that of Tamiflu. The prophylactic effect of Jin Qiao Tablets was stronger than the therapeutic effect, but there was no significant difference between them. Conclusion Jin Qiao Tablets have obvious effect against influenza A H1N1 virus in vivo.

The Chinese herbal Radix Scrophulariae is the main medicine for nourishing yin and reducing fire. It can be used to treat hyperthyroidism due to yin deficiency and fire hyperactivity, but its mechanism is not clear. The present study was aimed to explore the mechanism of Radix Scrophulariae treatment of hyperthyroidism due to yin deficiency and fire hyperactivity. The urine metabolomic approach was conducted using the method of UPLC-TOF-MS. The results showed that Radix Scrophulariae has good therapeutic effects on hyperthyroidism rat model of yin deficiency. After treatment with Radix Scrophulariae, through metabolic profiling and protocol analysis, 6 potential metabolic markers may be closely related with the treatment mechanism of Radix Scrophulariae on this disease, including proline betaine, estrone, thymidine, 5-hydroxyindoleacetic acid, cyclic AMP and L-dopa. The strongest metabolic pathways were associated with tryptophan metabolism, pyrimidine metabolism, tyrosine metabolism, purine metabolism and steroid hormone biosynthesis. The urine metabolomic approach can be applied to clarify the therapeutic mechanism of Radix Scrophulariae on hyperthyroidism rat of yin deficiency, and provide the theoretical basis for the clinical practice of Radix Scrophulariae on nourishing yin to reduce pathogenic fire.

Aim To build the model of the gene FKBP38(FK506 binding protein 38)conditional knock out in uterus and then investigate the effect on endometrial precancerous lesions and the underlying mechanism.Methods Transgenic mice whose FKBP38 gene was flanked with loxP were constructed by embryo microinjection. The conditional knockout of FKBP38 was obtained by breeding mice harboring two loxP sites in FKBP38(FKBP38

Aim To investigate the role of autophagy core protein Atg5 in maintaining epididymal physiological functions and sperm maturation. Methods The Atg5 conditional knockout mouse model of principle cells in the 4 - 5 segments of the epididymal caput was constructed by Cre/LoxP system. The mice were divided into three groups according to genotype: Atg5v(, Atg5u~ and Atg5~/~ . The pregnancy rate and litter sizes were recorded by mating experiments at the age of 8 weeks-old. Sperm motility, sperm counts were evaluated with computer-aided sperm analysis ( CASA) after 14 weeks of feeding. HE staining was conducted to observe the morphology of the epididymal caput. Western blot and immunofluorescence technologies were applied to verify the expression level of Atg5 and the impediment of autophagy after Atg5 conditional knock out. Results After tissue specific knockout of Atg5 in the 4-5 epididymal principle cells, the autophagy marker proteins, LC3- I and p62, were accumulated and autophagy was successfully blocked. There was no significant difference in the morphology of epididymal tissues of the three genotypes of mice, nor any statistically significant difference in sperm motility, sperm count, litter size and pregnancy rate. Conclusions Under normal conditions without external challenge, Atg5 conditional knockout led to autophagy blocking of epididymal caput, but had no effect on mouse epididymal function and sperm maturation process.

Objective:To investigate the protective effect of endogenous ω-3 polyunsaturated fatty acid(PUFA)against cisplatin-induced myelosuppression and the mechanism of reducing apoptosis in bone marrow nucleated cells using mfat-1 transgenic mice.Methods:The experimental animals were divided into 4 groups:wild-type mice normal control group,mfat-1 transgenic mice normal control group,wild-type mice model group and mfat-1 transgenic mice model group.The mice in the model group were injected intraperitoneally with 7.5 mg/kg cisplatin on day 0 and day 7 to construct a myelosuppression model,while the mice in the normal control group were injected intraperitoneally with an equal amount of saline,and their status was observed and their body weight was measured daily.Peripheral blood was taken after 14 day for routine blood analysis,and the content and proportion of PUFA in peripheral blood were detected using gas chromatography.Bone marrow nucleated cells in the femur of mice were counted.The histopathological changes in bone marrow were observed by histopathological staining.The apoptosis of nucleated cells and the expression level changes of apoptosis-related genes in the bone marrow of mice were detected by flow cytometry and fluorescence quantitative PCR.Results:Compared with wild-type mice,mfat-1 transgenic mice showed significantly increased levels of ω-3 PUFA in peripheral blood and greater tolerance to cisplatin.Peripheral blood analysis showed that endogenous ω-3 PUFA promoted the recovery of leukocytes,erythrocytes,platelets and haemoglobin in peripheral blood of myelosuppressed mice.The results of HE staining showed that endogenous ω-3 PUFA significantly improved the structural damage of bone marrow tissue induced by cisplatin.Flow cytometry and PCR showed that,compared with wild-type mice model group,the apoptosis rate of bone marrow nucleated cells in mfat-1 transgenic mice was significantly reduced(P＜0.001),and the expression of anti-apoptotic genes Bcl-2 mRNA was significantly increased(P＜0.01),while the expressions of pro-apoptotic genes Bax and Bak mRNA were significantly reduced(P＜0.001,P＜0.05).Conclusion:Endogenous ω-3 PUFA can reduce cisplatin-induced apoptosis in bone marrow nucleated cells,increase the number of peripheral blood cells and exert a protective effect against cisplatin-induced myelosuppression by regulating the expression of apoptosis-related genes.

Aim To express and purify rhα-Gal A with a 6 X His tag via using a serum-free expression system in high-density suspension culture of Chinese hamster ovary cells ( CHO-S) , and to verify the scavenging effect of rhα-Gal A on globular trisaccharide ceramide (Gb3 or GL3) . Methods The construction of recombinant protein expression vector, pcDNA4-GLA, was achieved by fusing the human α-galactosidase cDNA, gla, with 6 X His tag and artificial DNA synthesis. The expression plasmid was transfected into the suspended CHO-S to express rhα-Gal A and then purified. Following this procedure, we determined rhα-Gal A's expression, the enzymatic activity, and the glycosylation of the recombinant enzyme. Co-incubation with cultured cells was performed to examine whether rhα-Gal A could be taken up into the cells and effectively remove Gb3 substrates. Results rhα-Gal A was successfully expressed and purified after transiently transfecting pcDNA4-GLA into the suspended CHO-S, and the yield was up to (100 ±20. 6) mg • L

Objective To investigate the effect of cysteine protease inhibitor derived from S chistosoma japonicum(SjCys-tatin)on dextran sodium sulfate(DSS)-induced acute ulcerative colitis in mice.Methods Eighteen C57BL/6 mice were ran-domly divided into three groups:a control group treated with PBS(Group A),a DSS-induced-colitis group treated with PBS(Group B),and a DSS-induced-colitis group treated with SjCystatin(Group C).Colitis was induced in mice by giving 3％DSS orally for 7 days.During this period,the mice were daily injected with 10&mu;g of SjCystatin or PBS only as a control intraperitone-ally.The mice were monitored daily for their clinical manifestations and given scores based on disease activity index(DAI).The severity of colonic inflammation was monitored by the macroscopic score and pathological change.The cytokine profile including TNF-α,IL-4,IL-6 and IL-10 in the supernatants of colon homogenate was detected by ELISA.Results Compared with Group A(0.50 ± 0.28),the DAI score increased significantly in Group B(9.30 ± 1.30)(F=86.86,P<0.01),with remarkable path-ological damages seen in colon tissues.and the levels of TNF-α and IL-6 were(321.33±67.01)and(403.58 ±180.51)pg/mL.The DAI score significantly reduced in Group C(6.67±1.57)as compared to Group B(F=86.86,P<0.01),with improve-ments in the macroscopic and microscopic pathology in mouse colon specimens.As compared to Group B,the levels of TNF-α [(188.14 ± 40.14)pg/mL] and IL-6 [(209.71 ± 48.47)pg/mL] significantly decreased(F=17.46 and 9.89,both P<0.01).Con-clusion SjCystatin has a significantly inhibitory effect for alleviating DSS-induced acute ulcerative colitis in C57BL/6 mice.

Aim To investigate the role of metabolites of eicosapentaenoic acid (EPA) in promoting the transdifferentiation of pancreatic α cells to β cells. Methods Male C57BL/6J mice were injected intraperitoneally with 60 mg/kg streptozocin (STZ) for five consecutive days to establish a type 1 diabetes (T1DM) mouse model. After two weeks, they were randomly divided into model groups and 97% EPA diet intervention group, 75% fish oil (50% EPA +25% DHA) diet intervention group, and random blood glucose was detected every week; after the model expired, the regeneration of pancreatic β cells in mouse pancreas was observed by immunofluorescence staining. The islets of mice (obtained by crossing GCG