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Journal of Jiangsu University(Medicine Edition)

	2015 Vol.25 Issue.03 Published 2015-05-30

Article

	1


		2015 Vol. 25 (03): 1- [Abstract] ( 502 ) [HTML 1KB] [ PDF 322KB] ( 870 )

	185	Effect of noncoding RNA AsrC on the intracellular survival of Salmonella enterica serovar Typhi in macrophage
		ZHANG Qi, Yang-Ru-Yue, Xu-Rong,
		Objective: To investigate the influence of the non-coding RNA AsrC on the intracellular survival of Salmonella enterica serover Typhi in macrophage and its mechanism. Methods: The macrophages intracellular survival assay was performed in AsrC overexpressing strain and the control strain; qRT-PCR was carried out to determine the transcriptional variation of asrC, rseC, rpoE; the expression of LC3 was detected by Western blotting. Results: Compared with the control strain, infected in 12, 24 h, the AsrC in overexpressing strain showed lower intracellular survival in macrophage (both P<0.01), higher levels of asrC mRNAand rseC mRNA(P<0.05), but had no influence on rpoE(P>0.05) ; Western blot analysis revealed that the expression of LC3 had no significant difference between AsrC overexpressing strain and the control strain(P>0.05). Conclusion: Overexpression of AsrC might decrease the bacterial intracellular survival in macrophage, which might not be dependent on autophagy. ［Key words］Salmonella enterica serovar Typhi; non-coding RNA; AsrC; intracellular survival; autophagy
		2015 Vol. 25 (03): 185- [Abstract] ( 940 ) [HTML 1KB] [ PDF 3891KB] ( 950 )

	190	Mechanism of TGF-β1 mediated NF-κB signal pathway inhibiting myogenic differentiation of rhabdomyosarcoma
		JIANG Zhen-1, 2 , Chen-Wang-Wang-1, 2 , Cao-Dan-3,
		Objective: To investigate the mechanism of TGF-β1 mediated NF-κB signal pathway inhibiting myogenic differentiation of rhabdomyosarcoma(RMS). Methods: After transfected with TGF-β1 siRNAexpressing plasmid system,the expression level of Caspase-3 were detected by immunohistochemical staining. The apoptosis of RD cells was assessed by TUNEL assay,and myofilament were explored by transmission electron microscope (TEM).After coculturing with exogenous TGF-β1，the expression level of nuclear factor-kappa B(NF-κB) family members such as p65,p50,p52 and RelB in RD cells were tested by immunofluorescence. The expression level of NF-κB family members in rhabdomyosarcoma paraffin tissues were determined by Immunohistochemistry. Results: Compared with the control group, the expression of Caspase3 and the positive cells detected by TUNEL assay were increased significantly (P<0.05); TEM showed that structure of myofilament significantly increased in RD cells treated with TGF-β1 siRNAexpressing plasmid system.After 6 hours for co-culturing with exogenous TGF-β1(2 ng/mL), RD cells showed significantly higher positive rate and staining intensity of p65 and p50 than those in control group（P<0.05）.The high expression rate of p65 and p50 was found in TGF-β1high RMS than that in TGF-β1low group(P<0.05). Conclusion: Differentiation inhibition of rhabdomyosarcoma may be related with the TGF-β1 signal transduction mediated by p65 and p50, which is belong to the members of NF-κB family. ［Key words］rhabdomyosarcoma; TGFβ1； nuclear factorκB； myogenic differentiation
		2015 Vol. 25 (03): 190- [Abstract] ( 1709 ) [HTML 1KB] [ PDF 4644KB] ( 1445 )

	195	Analgesic effect of gastrodin on metastasizing in cancer-induced pain mouse model
		WANG Hua-1, Shao-Dong-Hua-2, Ma-Peng-3
		Objective: To observe the analgesic effect of gastrodin on metastasizing cancer-induced pain model mice. Methods: To establish the cancer induced pain model through injecting subcutaneously breast cancer cells into the mouse hind paw. Thirtyfive 8 weeks old female Balb/c mice were randomly divided into 5 groups: control group,solvent group, low dose gastrodin (90 mg/kg)，high dose gastrodin (180 mg/kg) and morphine group (10 mg/kg).At 11th days after tumor cells inoculation, thermal hyperalgesia test were measured respectively at 15,30,60,90,120,150 and 180 minutes after drug delivery; and compared to each other. Results: At the 90 minutes after drug delivery,mice thermal hyperalgesia threshold in low dose gastrodin group increased to (6.19±0.23)s from (3.63±0.33)s；mice thermal hyperalgesia threshold in high dose gastrodin group increased to (6.97±0.33)s from (3.83±0.14)s.And the increase amplitude of threshold were smaller than morphine group（P＜0.01，P＜0.05）. Conclusion: Gastrodin can produce analgesic action on cancerinduce pain and the analgesic effect of gastrodin weaker than morphine. ［Key words］gastrodin; cancerinduced pain; analgesia; morphine
		2015 Vol. 25 (03): 195- [Abstract] ( 1028 ) [HTML 1KB] [ PDF 2877KB] ( 1651 )

	199	NK/c-Jun signaling pathway contributes to mediate Epstein-Barr virus-encoded BARF1 gene up-regulating Bcl-2 expression in human gastric carcinoma cells
		ZHANG Yu-Qiong, Zhang-Xue-Yi, Xu-Mei-Qin
		Objective: To investigate the effect of the JNK/c-Jun signaling pathway on mediating Epstein-Barr virus-encoded BARF1 gene up-regulating Bcl-2 expression in human gastric carcinoma cells. Methods: Western blotting was performed to analyze the levels of JNK and c-Jun protein phosphorylations and of the total c-Jun and Bcl-2 protein expressions in BARF1stablyexpressing human gastric carcinoma cell line BGC823 (BGC-BARF1) under the control of pSG5 vector transfected one (BGC-SG). Results: The levels of the JNK and c-Jun protein phosphorylations, and of the c-Jun and Bcl-2 protein expressions were significantly increased in BGC-BARF1 compared with the BGC-SG controls, and blocked by special inhibitor of JNK-MAPK SP600125（all P< 0.05）. Conclusion: JNK/c-Jun signaling pathway contributes to mediate BARF1 gene up-regulating the Bcl-2 expression in human gastric carcinoma cells. ［Key words］BARF1; Epstein-Barr virus; gastric carcinoma; JNK; Bcl-2
		2015 Vol. 25 (03): 199- [Abstract] ( 1375 ) [HTML 1KB] [ PDF 2920KB] ( 1393 )

	203	Exosome derived from rat bone marrow mesenchymal stem cells inhibits the apoptosis in cisplatin-induced renal proximal tubular epithelial cells
		ZHU Yuan, Yang-Fang, Zhang-Jiao
		Objective: To investigate the effect of exosome derived from rat bone marrow mesenchymal stem cells on the apoptosis of cisplatin-induced renal proximal tubular epithelial cells. Methods: Primitive rat bone marrow mesenchymal stem cells were separated and cultured in vitro from which exosome were extracted and proofed. The injury of NRK52E cells was induced by 5 μmol/L cisplatin for 6 h and cultured for another 48 h using normal DMEM. 100 μg/mL exosome contained DMEM was used in restoration group for the 48 h after the cisplatin treatment, and normal NRK-52E cells with no treatment were used as control. TUNEL-FITC, flow cytometry and immunoblotting were used to detect the apoptosis in each group. Results: Compared with the control group, NRK-52E cells in cisplatin group grew into swollen ones with nucleus changed irregularly, the number of apoptotic cells was increased; the expression of apoptosis associated protein Bax was upregulated while Bcl-2 downregulated. However, in the exosome group, the phenomenon of swell and irregular nucleus in NRK-52E cells was alleviated. The number of apoptotic cells was decreased, with decreased expression of Bax and increased expression of Bcl-2. Conclusion: Exosome derived from rat bone marrow mesenchymal stem cells could suppress the apoptosis induced by cisplatin in NRK-52E cells. ［Key words］exosome; cisplatin; renal proximal tubular epithelial cell; injury; apoptosis
		2015 Vol. 25 (03): 203- [Abstract] ( 846 ) [HTML 1KB] [ PDF 4052KB] ( 1586 )

	207	Effect of oxymatrine on radiosensitivity of A549 cells and its possible molecular mechanism
		SHI Wei-Lin-1, Li-Jian-2, Lu-Jian-Bao-1
		Objective: To investigate the mechanism of oxymatrine in enhancing the sensitivity of lung cancer cells to radiation. Methods: The A549 cells were divided into control group, radiation group, drug group, drug plus radiation group according to treatment condition. The cell counting kit-8 assay was used to determine proliferation rate. The real-time quantitative PCR assays were conducted to measure DNA-PKcs and Ku80 mRNA expression levels at 24 h and 48 h following treatment with oxymatrine, or radiation, or oxymatrine plus radiation. Results: Oxymatrine significantly inhibited the proliferation of A549 cells in a dose and time dependent manner(both P＜0.05). IC50 of A549 cells for oxymatrine at 24 h and 48 h were 3.2 g/L and 2.4 g/L, respectively.The proliferation rates of A549 cells in the radiation group were decreased in a time and dose dependent manner (both P＜0.05). At the same time points, the cell proliferation rates in the drug plus radiation group were dropped markedly when compared with the radiation group(P＜0.05). Additionally, the relative expression levels of DNA-PKcs and Ku80 mRNA were increased in a dose and time dependent manner in the radiation group(both P＜0.01). The relative expression levels of DNA-PKcs and Ku80 mRNA in the drug plus radiation group were reduced significantly as compared with in the radiation group(P＜0.01). Conclusion: Oxymatrine enhanced the sensitivity of A549 cell to radiation, at least partially, by inhibiting DNA-PKcs and Ku80 mRNA expression and damage of DNA damage repair mechanisms. ［Key words］lung cancer; oxymatrine; radiosensitivity; DNA dependent protein kinase
		2015 Vol. 25 (03): 207- [Abstract] ( 800 ) [HTML 1KB] [ PDF 3565KB] ( 1377 )

	212	A viral vector expressing secretory β sheet breaker peptide inhibited Aβ neuronal toxicity
		WANG Bing, Liu-Qing-Qing, Xia-Chun-Lin
		Objective: To observe biological activity of secretory β sheet breaker peptide (BSB) expressed via a viral vector in cultured hippocampal neurons. Methods: We constructed the recombinant adeno-associated virus (rAAV) encoding fusion gene of neurotrophin-4(NT-4) signal peptide and BSB by molecular cloning technique. The virus were produced in 293 packaging cell lines of the AAV and purified by sucrose gradient centrifugation. The viral titer was determined by dot blot hybridization. The biological effects of expressive BSB in vitro were observed by MTT assay and electron microscopy. Results: The pSSHG/NT4BSB plasmid was constructed successfully. The physical titer of recombinant AAV was 1×1011-1×1012 virions/mL after purification. The BSB, secretive expression in AAV transfected cultured hippocampal neuron, inhibited Aβ fibrosis aggregation and significantly decreased the neurotoxicity of Aβ in cultured hippocampal neuron. Conclusion: AAV vector mediated secretive expression of BSB peptide displayed effective biological effect in vitro cultured neurons. ［Key words］Alzheimer′s disease; βsheet breaker peptide; signal peptide; adenoassociated virus
		2015 Vol. 25 (03): 212- [Abstract] ( 961 ) [HTML 1KB] [ PDF 3955KB] ( 1525 )

	217	Effect of two image reconstruction algorithms on the quantification analysis of experimental animal tumor model with 18F-FDG PET/CT
		LIU Ping-An-1, Wan-Liang-Rong-2, Huang-Gang-2
		Objective: To compare the effect of image reconstruction algorithms on the quantification analysis of experimental animal tumor model with 18F-FDG PET/CT. Methods: Dynamic scans with18F-FDG PET/CT were performed in five New Zealand rabbits bearing VX2 tumor in fore limbs, tumor dynamic images were obtained and reconstructed with both filtered back projection method (FBP) and ordered-subset expectation maximization algorithm (OSEM). Quantification parameters of Ki and SUVmax were calculated and compared. Results: More noise presented in the images reconstructed by FBP than that by OSEM. Ki and SUVmax calculated by both reconstruction algorithms correlated well (r≥0.95); Ki and SUVmax calculated from images reconstructed by OSEM（Ki：0.03±0.013；SUVmax: 4.89±1.69）were significantly higher than that by FBP(Ki:0.025±0.011; SUVmax: 4.14±1.42）, all P<0.05. Conclusion: Reconstruction algorithms had effect on the quantification measures，and parameters calculated from image reconstructed by OSEM was higher than that by FBP. ［Key words］tomography; dynamic PET; FDG; image reconstruction
		2015 Vol. 25 (03): 217- [Abstract] ( 1056 ) [HTML 1KB] [ PDF 2972KB] ( 1531 )

	221	Effects and mechanism of Toll like receptor-4 in osteoclastgenesis of Effects and mechanism of Toll like receptor-4 in osteoclastgenesis of stem cells in co-culture system with osteoblast stem cells in coculture system with osteoblast
		ZHU Hua-Rong-1, Xu-Xiao-Feng-1, Chen-Qi-1, WU Wei-2
		Objective: To investigate the effect of Toll like receptor (TLR)-4 to osteoclastgenesis of stem cells to osteoclasts in co-culture system with osteoblast, and to research the probably mechanism. Methods: Bone marrow mesenchymal stem cells (BMSCs) of rat were abstracted with percoll density gradient centrifugation method. Surface maker CD34, CD44, CD54, and CD90 were identified with flow cytometry. BMSCs (upper cell)-osteoblasts (lower cell) co-culture system was established with Transwell cells, BMSCs were induced to osteoblasts in this system. Co-culture system were divided into TLR-4 activated group and blank group. Cells in TLR-4 activated group were activated by LPS (10 ng/mL), cells in blank group were treated with PBS of the same volume. After 6 days culture, tartrate resistant acid phosphatase (TRAP) staining was performed to examine osteoclasts in the lower cell. Osteoclasts numbers in each group were counted and compared. Meanwhile, western blotting was performed to investigate the expression of receptor activator for nuclear factor-κB ligand (RANKL). Results: BMSCs of rat were abstracted with density gradient centrifugation method. Surface maker CD34 was negative, while CD44, CD54, and CD90 were positive. TRAP staining positive cell was osteoclasts. Osteoclasts numbers in each group were counted, it was obviously higher in TLR-4 activated group than that in blank group (t=13.556, P<0.05). Results of western blotting showed RANKL expression in TLR-4 activated group was higher than that in blank group(t=17.630, P<0.05). Conclusion: In “BMSCs-osteoblasts” co-cultured system, activation of TLR-4 is benefit to the osteoclastgenesis of stem cells. The promotion of TLR-4 to the secretion of RANKL may be the mechanism. ［Key words］stem cells; Toll like receptor4; osteoclast; receptor activator for nuclear factorκB ligand
		2015 Vol. 25 (03): 221- [Abstract] ( 1152 ) [HTML 1KB] [ PDF 5381KB] ( 1382 )

	225	Correlation of Caveolin-1 expression with microlymphatic density in colorectal adenocarcinoma tissues and its clinical significance
		GUO Fei, Xue-Jun, Wu-Xue-Liang
		Objective: To study the expression of Caveolin-1 in colorectal adenocarcinoma tissues and its correlation with microlymphatic density (LMVD)，and to investigate the clinical pathological prognostic significance of Caveolin-1 and LMVD in patients with colorectal cancer. Methods: The expression of Caveolin-1 and LMVD in 45 specimens of normal colorectal tissues, and 90 specimens of colorectal adenocarcinoma tissues were detected by immunohistochemistry technique. We also investigated the correlation between their expression and the clinicopathologic tissues features. Results: The positive rates of Caveolin-1 in colorectal adenocarcinoma (73.33%) were significantly higher than those in normal colorectal tissues (11.11%, P<0.01); The positive rates of LMVD in colorectal adenocarcinoma tissues (18.25±2.36) were significantly higher than those in normal colorectal tissues (3.14±1.58, P<0.01). Expression of Caveolin-1 and LMVD value was correlated with liver metastasis, lymph node metastasis, TNM stage and the depth of tumor invasion.The mean LMVD in group with Caveolin-1 positivity expression was significantly higher than in that with Caveolin-1 negativity expression. Conclusion: The expression of Caveolin-1 in colorectal cancer was closely related to lymphatic microvessel of tumor, and its expression and LMVD were independent prognostic indicators for patients with colorectal cancer. ［Key words］colorectal neplasms; Caveolin1; microlymphatic density; immunohistochemistry
		2015 Vol. 25 (03): 225- [Abstract] ( 690 ) [HTML 1KB] [ PDF 3202KB] ( 1331 )

	229	Metaanalysis on the association of serum adiponectin level and polycystic ovarian syndrome risk in Chinese Han women
		JIN Yan-1, Wang-Kun-2, Liu-Shan-Shan-3
		Objective: To evaluate the association of serum adiponectin concentrations with polycystic ovarian syndrome (PCOS) in Chinese Han women. Methods: By formulating the retrieval strategy, papers related to the association of serum adiponectin concentrations with PCOS in Chinese Han women published before the December in 2014 were retrieved from CNKI, VIP, Wanfang, CBM, PubMed, EMbase and so on. The paper screened was based on the inclusion/exclusion criteria. RevMan 5.0 software was used for the Meta-analysis. Results: Seventeen studies were included, with 1 045 cases of PCOS and 659 non-PCOS subjects. Meta-analysis results showed that (1) Patients with PCOS had lower serum adiponectin concentrations than the control group(WMD=-4.10，95％CI=-5.79--2.40，Z=4.73，P<0.000 01); (2) the PCOS patients with normal weight (BMI<25 kg/m2) had lower serum adiponectin concentrations than normal weight control group (WMD=-2.00，95％CI=-2.28--1.71，Z=13.79，P<0.000 01); (3) the over weight/obesity PCOS patients(BMI≥25 kg/m2) had lower serum adiponectin levels than both the normal weight PCOS group (WMD=1.63，95％CI=0.91-2.36，Z=4.41，P<0.000 1), and the over weight/obesity control group (WMD=-1.64，95％CI=-2.06--1.22，Z=7.61，P<0.000 01), respectively. Conclusion: Adiponectin concentration was significantly lower in Chinese Han women with PCOS, which might be used as an independent factor of overweight/obesity for PCOS. ［Key words］adiponectin; polycystic ovarian syndrome; Metaanalysis
		2015 Vol. 25 (03): 229- [Abstract] ( 794 ) [HTML 1KB] [ PDF 6195KB] ( 1328 )

	236	Clinical utility of plasma miRNA-155 and miRNA-625 detection in the diagnosis of non-small cell lung cancer
		FEI Ling-1, Li-Jian-1, Zhu-Li-Huan-1, Lin-Jie-2
		Objective: To evaluate the clinical utility of plasma miRNA-155 and miRNA-625 expression as molecular markers in the diagnosis of non-small cell lung cancer(NSCLC). Methods: Fluorescent quantitative RT-PCR was used to detect the expressions of miRNA-155 and miRNA-625 in plasma samples from 49 patients with NSCLC and 55 patients with benign lung disease(BLD). Receiver operating characteristics(ROC) curves of the two miRNAs were constructed to assess diagnostic power for NSCLC. The sensitivity and specificity of plasma miRNA-155 and miRNA-625 expression for diagnosing NSCLC were calculated, and compared with serum carcinoembryonic antigen(CEA). Results: The plasma miRNA-155 levels in NSCLC patients were significantly higher than those in BLD patients(P=0.000), whereas the plasma miRNA-625 levels in NSCLC patients were significantly lower than those in BLD patients(P=0.000). For patients with NSCLC, the plasma miRNA-155 and miRNA-625 expression levels were significantly correlated with tumor clinical (TNM) stage(P=0.008 and P=0.025, respectively). In addition, plasma miRNA-625 levels were also correlated with NSCLC patient gender (P=0.024). No correlation was found between the plasma miRNA-155 and miRNA-625 levels and other clinicopathologic characteristics. According to the ROC curve analysis, cutoff values of plasma miRNA-155 and miRNA-625 levels in differentiating NSCLC from BLD were defined as 2.736 and 0.699, respectively. For the diagnosis of NSCLC, the sensitivity and specificity was 75.5% and 74.5% for miRNA-155, 73.4% and 70.9% for miRNA-625, respectively. The sensitivity of CEA was inferior to miRNA-155 and miRNA-625(P=0.004 and P=0.007), although the CEA specificity was similar to the two miRNAs for diagnosing NSCLC. Conclusion: Plasma miRNA-155 and miRNA-625 expression might be used as the molecular markers for the diagnosis of NSCLC. ［Key words］nonsmall cell lung cancer; miRNA155; miRNA625; plasma; diagnosis
		2015 Vol. 25 (03): 236- [Abstract] ( 958 ) [HTML 1KB] [ PDF 4516KB] ( 1589 )

	241	Analysis of clinicopathological and prognostic characteristics in hepatolithiasisassociated intrahepatic cholangiocarcinoma
		SU Xin-Cheng-1, Pang-Shu-Jie-1, Yang-Ning-1, Xu-Zhu-Ding-2
		Objective: To describe the difference between the hepatolithiasis(HL)-positive intrahepatic cholangiocarcinoma(ICC) and hepatolithiasis-negative intrahepatic cholangiocarcinoma, and examine the prognosis based on our data. Methods: A total of 296 patients with ICC underwent the first curative resection were involved in our research, in whom 38 were with hepatolithiasis , the rest 258 were without. Data of sixteen clinicopathological features, recurrence free survival(RFS) rate and overall survival(OS) rate were collected and examined retrospectively. Results: About the sixteen clinicopathological features, the age of the HL-associated ICC patients tend to be older and with more female gender. The HL-positive group tends to have a higher level of CA19-9, ALP,r-GT. Meanwhile, the HL-positive ICC was more likely to be accompanied with satellite lesions, lymph node metastasis, nerve invasion. Moreover, the HL-positive ICC tends to have a more progressive TNM staging. The actual 1-,3-,5-year recurrence free survival(RFS) rate in the HL-positive group was 19.8%,2.6%,0% and in the HL-negative group was 46.9%，26.4%, 20.9%.The 1-, 3-, and 5-year overall survival rate was 39.5%,7.9%,0% in the HL-positive group and in the HL-negative group was 67.8%,38.0%,26.4%. Conclusion: The HLpositive ICC has a more progressive degree and a poorer prognosis compared with the HL-negative ICC. So that the HL-positive ICC were prone to form the lymph node metastasis, and lymphadenectomy in the hepatoduodenal ligament was suggested. ［Key words］intrahepatic cholangiocarcinoma; hepatolithiasis; survival
		2015 Vol. 25 (03): 241- [Abstract] ( 963 ) [HTML 1KB] [ PDF 3663KB] ( 1333 )

	246	onstruction and in vitro expression of Toxoplasma gondii SAG1, ROP18 eukaryotic expression plasmids
		WU La-Mei-1, Wu-Ting-1, Wu-Liang-1
		Objective: To construct two recombinant eukaryotic expression plasmids containing the surface antigen (SAG)1 and rhoptry protein (ROP)18 gene of Toxoplasma gondii, and express SAG1 and ROP18 proteins in the eukaryotic system. Methods: The gene fragments encoding SAG1 and ROP18 were amplified by PCR with special primers from Toxoplasma gondii genomic DNA respectively, and were cloned into pTG19-T vector. The right gene fragments were subcloned into p3×FLAGMycCMVTM24 vector to construct the eukaryotic expression plasmids p3×FLAG-Myc-CMVTM-24-SAG1 and p3×FLAG-Myc-CMVTM-24-ROP18. The recombinant plasmids were transfected into human renal epithelial 293-T cell lines respectively, and the expression of SAG1 and ROP18 in transfected cells were detected by RT-PCR and Western blotting. Results: The gene fragments encoding SAG1, ROP18 were 1 011 bp and 1 665 bp respectively, which were consistant with the expected size. The analysis of enzyme digestion, PCR and sequencing showed that the recombinant eukaryotic expression plasmids p3×FLAG-Myc-CMVTM-24-SAG1, p3×FLAG-Myc-CMVTM-24-ROP18 were constructed correctly. RT-PCR and Western blotting analysis of cells transfected SAG1, ROP18 gene displayed positive bands. Conclusion: Recombinant eukaryotic expression plasmids of SAG1, ROP18 have been successfully constructed, the recombinant plasmids can express SAG1, ROP18 proteins in a eukaryotic system. ［Key words］Toxoplasma gondii; SAG1; ROP18; eukaryotic expression plasmid
		2015 Vol. 25 (03): 246- [Abstract] ( 1011 ) [HTML 1KB] [ PDF 4627KB] ( 1403 )

	251	Prokaryotic expression and polyclonal antibody preparation of rhoptry protein 16 from Toxoplasma gondii
		LIU Yuan-1, Yang-Hua-1, Wu-Liang-1, CAO Jian-Ping-2
		Objective: To clone and construct a recombinant expression plasmid of rhoptry protein（ROP）16 from tachyzoites of Toxoplasma gondii(T. gondii) RH strain in Escherichia coli, and to determine and locate the expression of ROP 16 in tachyzoites. Methods: The ROP16 gene was amplified from genomic DNA of T. gondii RH strain and cloned into the plasmid pET-32a(+). The expression of the recombinant pET32a-ROP16 was induced in E.coli Rosetta strain, and ROP16 was purified by cutting the gel slices with KCl stain. The antiROP16 polyclonal antibody was produced in rabbit, and ROP16 in tachyzoites was analyzed by Western blotting and indirect immunofluorescence assay (IFA) respectively. Results: The recombinant plasmid pET32a-ROP16 was successfully expressed and purified, and the anti-ROP16 polyclonal antibody was obtained. The 74 000 specific band of ROP16 was detected by Western blotting, and the ROP16 was distributed in the cytoplasm of tachyzoites by IFA. Conclusion: The anti-ROP16 polyclonal antibody prepared by recombinant ROP16 can detect and locate the expression of ROP16 in tachyzoites of T. gondii. ［Key words］Toxoplasma gondii; ROP16; prokaryotic expression; KCl stain
		2015 Vol. 25 (03): 251- [Abstract] ( 789 ) [HTML 1KB] [ PDF 3821KB] ( 1309 )

	256	Preparation and pharmacokinetics in rat of vinorelbine biartrate liposomes by ammonium sulfate gradient method
		ZHENG Zhao-Lei-1, Li-Ya-Ping-1, 2 , Chen-Ling-Li-2
		Objective: To prepare vinorelbine bitartrate（NVB） liposomes by ammonium sulfate gradient method, and to study the properties and pharmacokinetics of NVB liposomes. Methods: NVB liposomes were prepared by ammonium sulfate gradient method.The mean diameter of liposomes was determined by dynamic light scattering. Encapsulation efficiency was determined by sephadex column, pharmacokinetics was evaluated in SD rats. Results: Mean particle size and encapsulation efficiency of NVB liposomes were （88.7±11.2）nm(n=3) and （90.71±0.16）%（n=3）.In addition, less than 20% of NVB liposomes was released in 48 hours. The main pharmacokinetic parameters of NVB liposomes were as follows: t1/2=（5.38±0.29）h andAUC _0→t=（98.11±0.12）mg·h-¹·L^-1. Conclusion: NVB liposomes exhibited high encapsulation efficiency. The pharmacokinetics of NVB liposomes showed prolonged elimination halflife and increased bioavailability. ［Key words］vinorelbine biartrate; liposomes; ammonium sulfate gradient method; pharmacokinetics
		2015 Vol. 25 (03): 256- [Abstract] ( 1269 ) [HTML 1KB] [ PDF 3554KB] ( 1627 )

	260	Ingredient analysis and antioxidant activity of water extract of Abelmoschus esculentus tea
		ZHOU Zhao-Xiang-1, Bai-Shi-Qi-2, Zou-Ye-3
		Objective: To analyze nutritional ingredient and active ingredient of Abelmoschus esculentus tea, and study on antioxidant activity of water extract. Methods: To analyze nutritional ingredient and active ingredient of Abelmoschus esculentus tea with GB method and agriculture standard method. Scavenging effects of water extract on DPPH and OH were determined. Results: The content of ash , crude protein, crude fat and total carbohydrate were 15.32%, 27.58%, 2.27% and 12.06% of dried sample,respectively. The amino acid analysis results showed that Abelmoschus esculentus tea contained many kinds of essential amino acid. The trace element analysis results showed that Abelmoschus esculentus tea contained K,Ca,Mg,Fe,et al. The active ingredient flavonoids and polysaccharide were 1.36% and 4.70%, respectively. The IC50 of water extract scavenging DPPH and OH were 156.8 mg/L and 674.5 mg/L, respectively. Conclusion: Abelmoschus esculentus tea has good nutritional value and antioxidant effect which can provide a new green health drinks for consumers. ［Key words］Abelmoschus esculentus tea; nutritional ingredient ; flavonoids; polysaccharide; antioxidant activity
		2015 Vol. 25 (03): 260- [Abstract] ( 1106 ) [HTML 1KB] [ PDF 2237KB] ( 1508 )

	263	Rapid analysis of active ingredients in Panax notoginseng using surface enhanced Raman spectroscopy
		WEI Cheng-Hua, Chen-Juan, Xu-Man-Ling
		Objective: To establish surfaceenhanced Raman spectroscopy (SERS) method for rapid analysis of active ingredients in Panax notoginseng． Methods: Panax notoginseng was pretreated simply，then we tested the powder，water and alcohol extracts of Panax notoginseng by SERS respectively．We also got the spectrum of single active ingredient of the alcohol extracts combined with TLC，and analyzed these spectrum． Results: The powder，water and alcohol extracts of Panax notoginseng only could obtain weak signals of SERS directly，but the single active ingredient of Panax notoginseng separated by TLC technology could obtain its SERS spectra ideally． Conclusion: The method established in this article for rapid analysis of Panax notoginseng was efficient，rapid，specific，and high sensitivity， it can be extended to quality control and analysis in the identification of other herbal medicines．［Key words］surfaceenhanced Raman spectroscopy； Panax notoginseng； rapid analysis
		2015 Vol. 25 (03): 263- [Abstract] ( 828 ) [HTML 1KB] [ PDF 5471KB] ( 1985 )

	268
		ZHU Chang-Yun, Ma-Jian-Guo

		2015 Vol. 25 (03): 268- [Abstract] ( 623 ) [HTML 1KB] [ PDF 2022KB] ( 1537 )

	271
		ZHOU Lei-1, Yan-Jin-Chuan-2, * , Yu-Wen-Min-1, Zhang-Xiao-Yong-1

		2015 Vol. 25 (03): 271- [Abstract] ( 660 ) [HTML 1KB] [ PDF 2155KB] ( 1245 )

	274
		HU Yue-1, Gao-Yi-Na-1, Chen-Bao-Ding-2, HUANG Run-Sheng-3, Yu-Li-1*, Zhang-Jian-Quan-4

		2015 Vol. 25 (03): 274- [Abstract] ( 981 ) [HTML 1KB] [ PDF 2162KB] ( 1873 )

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