Object To establish a tissue culture and rapid propagation system for the medicinal plant Iphigenia indica Kunth. Methods Clusters of seedlings and protocorms and induction were studied on MS media with different parts of the plant, such as corms, stems, leaves and root tips as explants. Results The proper media for protocorm inducing, propagation and rooting were found in this paper, rapid propagation system of I. indica was established and the plant could put in large scale production. Conclusion Sprout can be successfully induced from corm and stem on MS media. MS+6-BA 2 mg/L+NAA 0.5 mg/L can induce both sprout and protocorm, and fit for large scale production. 1/2 MS+IBA 0.5 mg/L+NAA 0.2 mg/L is good for rooting.

Objective Accodring to the guideline of Reference Intervals in the Clinical Laboratory(ISO15189) ,to establish the reference range of Cu ,Fe ,Zn ,Ca and Mg in the peripheral blood of preschool children in Suzhou .Methods Using atomic absorp-tion spectrometry to detect the levels of Cu ,Fe ,Zn ,Ca and Mg in peripheral blood of 3 992 Kindergarden children in Suzhou from Apr to May 2014 ,according the age ,the children were divided into three groups ,1 511 cases in 3 to 4 years old group including 750 boys ,761 girls ,1 169 cases in 4 to 5 years old group including 570 boys ,599 girls ,1 312 cases in 5 to 6 years old group inclu-ding 604 boys 708 girls .According to NCCLS's guideline C28 -A2 to define the reference range .Results The concentration of copper ,iron ,zinc ,calcium ,magnesium had no significant differences in different gender and age groups(P>0 .05) ,the reference ranges in peripheral blood of the preschool children in Suzhou were Cu 11 .47-32 .53 umol/L ,Fe 6 .86-9 .80 mmol/L ,Zn 64 .46-133 .02 umol/L ,Ca 1 .47-2 .09 mmol/L ,Mg 1 .21 -1 .75 mmol/L .Conclusion There are regional difference on the levels of Cu ,Fe ,Zn ,Ca ,Mg in children's peripheral blood ,each region should to establish the local reference range follow the guideline .

Objective To evaluate the value of technetium-99m methoxy isobutyl isonitrile (99Tcm-MIBI)tomography and integrated CT in preoperative localization diagnosis of secondary hyperparathyroidism (SHPT).Methods 28 patients with SHPT were selected.15 minutes and 90 minutes (delayed images)after intravenous administration of 99Tcm-MIBI 370 MBq,static anterior planar images of the neck and chest were obtained.99Tcm-MIBI tomography and integrated CT scan were acquired after the delayed imaging.The imaging findings were compared to the pathological results.Results The detection rate of SHPT lesions by 99Tcm-MIBI dua-phase imaging and 99Tcm-MIBI tomography and registration with integrated CT scan was 53.3％ and 70.7％ respectively.The difference had statistical significance(X2 =5.903,P＜0.05).The fusion imaging with integrated CT can provide detailed anatomy data and more information for surgical doctors.Conclusion 99Tcm-MIBI tomography and registration with integrated CT scan can improve the detection rate of SHPT compared with 99Tcm-MIBI dua-phase imaging,provide more detailed anatomy data,and has high clinical value in preoperative localization and diagnosis of SHPT.

BACKGROUND: The particular bystander effect of suicide gene can remarkably kill tumor cells. Meanwhile, it can be used together with radiotherapy as well as immune gene therapy, and overcome the defect of low gene transduction efficiency.Cytosine deaminase (CD) can generate a powerful bystander effect.OBJECTIVE: To observe the effect of a eukaryotic expression plasmid plRES2-AcGFP1-CD mediated by liposome transfected into bone marrow mesenchymal stem cells (BMSCs) and its gene expression.DESIGN, TIME AND SETTING: A cytologic experiment at genetic level was performed at Research and Development Center of Stem Cell and Tissue Engineering, Dalian University of Technology from May to December 2007.MATERIALS: A total of 6 C57BL mice of SPF degree and weighing 18-20 g were used for separation and culture of BMSCs.Escherichia coli DH5a was provided by Research and Development Center of Stem Cell and Tissue Engineering, Dalian University of Technology. Lipofectamine~(TM) 2000 was provided by Invitrogen, China.METHODS: The DNA plasmids were extracted from transformed Escherichia coli DH5a. plRES2-AcGFP1-CD plasmid was identified by BamHI/Xhol double digestion. The BMSCs derived from mouse femur and tibia were cultured and purified by adhesion method in vitro. Signal cell suspension prepared by BMSCs of the third generation was cultured by adding fluorescence-labeled CD44, CD45, CD90 and CD105 antibodies. BMSCs of the third generation were transfected by LipofectamineTM 2000 mediation.MAIN OUTCOME MEASURES: Identification of recombinant plasmids; the expressions of surface markers on BMSCs were detected by flow cytometry; the expressions of cytosine deaminase gene were observed at 36 and 48 hours after transfection under fluorescent inverted microscope.RESULTS: After agarose gel electrophoresis, a band appeared at 1.0-1.5 kb of the digested products of plRES2-AcGFP1-CD plasmids, and the band was accorded with the length of cytosine deaminase gene in the length. Flow cytometry demonstrated that the cells were negative for CD45 but positive for CD44, CD90 and CD105. Fluorescent inverted phase contrast microscope suggested that at 36 hours after plRES2-AcGFP1-CD gene transfection an expression of green fluorescent protein was found in the BMSCs; additionally, at 48 hours after transfection, the expression of green fluorescent protein remained and the intensity was remarkably increased.CONCLUSION: The liposome-mediated plRES2-AcGFP1-CD gene successfully expressed in BMSCs, and the expression peaked at 48 hours after transfection.

Objective To evaluate the effect of dexmedetomidine on lipid peroxidation during lung ischemia-reperfusion (I/R) in rats.Methods Fifty-four pathogen-free healthy male Wistar rats,weighing 250-350 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (S group),I/R group,and dexmedetomidine group (Dex group).In group Dex,dexmedetomidine 5 μg/kg was injected via the caudal vein once a day for 2 consecutive days.The equal volume of normal saline was given in S and I/R groups.At 30 min after administration on 2nd day,lung I/R was produced by clamping the left hilum of lung for 45 min followed by 120 min of reperfusion in I/R and Dex groups.At 45 min of ischemia,and 60 and 120 min of reperfusion,6 rats selected from each group were sacrificed,and the left lungs were removed for examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),malondialdehyde (MDA) content and superoxide dismutase (SOD) activity.Results Compared with S group,the SOD activity was significantly decreased,and the MDA content and W/D ratio were significantly increased at 60 and 120 min of reperfusion in I/R and Dex groups (P＜0.05).Compared with I/R group,the SOD activity was significantly increased,and the MDA content and W/D ratio were significantly decreased at 60 and 120 min of reperfusion in Dex group (P＜0.05).The pathological changes of lungs were significantly attenuated in Dex group as compared with I/R group.Conclusion Dexmedetomidine mitigates lung I/R injury through inhibiting lipid peroxidation in rats.

Background and purpose:The effect of TPF (docetaxel, cisplatin and 5-lfuorouracil) induction chemotherapy plus concurrent chemoradiotherapy in locoregionally advanced nasopharyngeal carcinoma is unclear. This study aimed to compare the outcomes and tolerance of neoadjuvant chemotherapy with TPF versus cisplatin and 5-lfuorouracil (PF) followed by concurrent chemoradiotherapy in locoregionally advanced nasopharyngeal carcinoma patients.Methods:Patients with locoregionally advanced nasopharyngeal carcinoma were randomly divided into 2 groups: Group TPF and Group PF. Group TPF: One hundred and sixteen nasopharyngeal carcinoma patients received TPF consisting of docetaxel at 60 mg/m2 on day 1, cisplatin at 60 mg/m2 on day 1, and 5-lfuorouracil at a dose of 750 mg/m2by 24 h continuous infusion for 5 days for 3 cycles with a 21 day interval; Group PF: One hundred and sixteen nasopharyngeal carcinoma patients received PF consisting of cisplatin at 80 mg/m2 on day 1, and 5-lfuorouracil at a dose of 750 mg/m2by 24 h continuous infusion for 5 days for 3 cycles with a 21 day interval. After the completion of neoadjuvant chemotherapy, all the patients received intensity modulated radiation therapy (IMRT) with concomitant chemotherapy consisting of 2 cycles of cisplatin at 80 mg/m2 on day 1 and day 22. The prescribed doses were 6 810 cGy at 2.27 Gy/fraction to the gross tumor volume (GTV) with 5 daily fractions per week for 6 weeks. The acute toxicity and tumor response rate (RR), including complete response (CR) and partial response (PR), were evaluated. Addition-ally, the 5-year progress-free survival (PFS) rates and overall survival (OS) rates were further evaluated.Results:RR of Group TPF was higher than that of group PF at the end of neoadjuvant chemotherapy and within 13 weeks of the completion of concurrent chemoradiotherapy. The median recurrence time of TPF group was 2.98 years, and the 5-year PFS was 84.48%. The median recurrence time of PF group was 2.32 years, and the 5-year PFS was 82.75%. There was no statistically signiifcant difference between the 2 groups (P=0.458). The 5-year OS of TPF group was 87.06%, and for the PF group was 85.34%. There was no statistically signiifcant difference between the 2 groups (P=0.274). The incidence of leukopenia, thrombocyte penia, liver and kidney damage, diarrhea and mucosa necrosis in TPF group were signiifcantly higher than those in PF group (P<0.001). TheⅢ andⅣ degrees adverse reactions in TPF group were sig-niifcantly higher than those in PF group (P<0.001).Conclusion:TPF induction chemotherapy was not superior to the PF regimen for locoregionally advanced nasopharyngeal carcinoma patients. It should not be recommended in terms of more acute toxicity.

Objective To evaluate the effect of oxycodone postconditioning on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty pathogen-free healthy adult male Sprague-Dawley rats,weighing 200-300 g,were randomly divided into 4 groups (n =10 each) using a random number table:sham operation group (group S),myocardial I/R group (group Ⅰ),oxycodone postconditioning group (group O),and selective protein kinase C inhibitor chelerythrine group (group CH).Myocardial ischemia was induced by 30 min occlusion of the left anterior descending branch of the coronary artery,followed by 120 min reperfusion.In group S,the left anterior descending branch of the coronary artery was only exposed but not ligated.In group CH,chelerythrine 5 mg/kg was injected intravenously and slowly via the jugular vein before ligation which was performed immediately after administration.In O and CH groups,oxycodone 0.5 mg/kg was injected intravenously and slowly via the jugular vein at 2 min before reperfusion.Arterial blood samples were taken at 120 min of reperfusion to detect the levels of cardiac troponin Ⅰ (cTnI) and creatine kinase-MB (CK-MB) in serum.The hearts were removed after the animals were sacrificed to measure the myocardial infarct size by TTC staining.Results Compared with group S,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in Ⅰ,O and CH groups (P＜0.05).Compared with group Ⅰ,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly decreased in O and CH groups (P＜0.05).Compared with group O,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in group CH (P＜0.05).Conclusion Oxycodone postconditioning can mitigate myocardial I/R injury in rats,and the mechanism is partially related to activation of protein kinase C signaling pathway.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are easily isolated and amplified, and facilitate the exogenous gene transfer and expression. In the human medicine, it is believed that BMSCs are ideal therapeutic cells and target cells in the gene therapy.OBJECTIVE: To investigate liposome-mediated cytosine deaminase (CD) gene transfecting rabbit BMSCs and its gene expression. DESIGN: A single sample observation. SETTING: Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University.MATERIALS: This study was performed at in the Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University from March 2006 to June 2007. New Zealand big-ear white rabbits of either gender, weighing 2.0-2.5 kg, with the age of 5 months old, were included in this study. METHODS: The CD gene was obtained from E.coli JM109 DNA by polymerase chain reaction (PCR). The fragment was cloned into pMD19-T vector. After restriction enzyme BamHI/XhoI digestion analysis and DNA sequence analysis, pIRES2-AcGFP1-CD eukaryotic expression plasmid was constructed. Meanwhile, BMSCs were harvested, cultured and identified. After enzyme digestion of eukaryotic expression plasmid, the rabbit BMSCs were transfected by Lipofectamine 2000-mediated method. Twenty-four hours after transfection, expression of green fluorescent protein was observed under an inverted fluorescent microscope. MAIN OUTCOME MEASURES: Construction of eukaryotic expression plasmid and identification of CD gene-transferred BMSCs. RESULTS: CD gene was cloned and connected to eukaryotic expression plasmid with green fluorescence. Twenty-four hours after transfecting rabbit BMSCs, it was found under an inverted microscope that under the excitation of 488 nm blue light, green fluorescence appeared in the pIRES2-AcGFP1-CD and pIRES2-AcGFP1 empty-plasmid transfected BMSCs, but not in the non-transfected ones. It indicates that CD gene successfully transferred BMSCs. CONCLUSION: BMSCs are ideal vectors in the CD gene therapy.

Objective To observe the effects of 2 Gy γ-ray irradiation on regulatory T cells (Tregs) and Th17 cells and immune balance of mice.Methods A total of fifty C57BL/6 male mice were randomly divided into two groups,the irradiated group exposed to 2 Gy of whole body γ-ray irradiation,and the control group sham-irradiated.At 1,3,7,14 and 28 d after radiation,changes of peripheral haemogram were detected respectively and Tregs in peripheral blood,thymus and spleen and Th17 cells in spleen were analyzed by flow cytometry.Results Compared with control group,the number of peripheral blood white cells (WBC) and lymphocyte in irradiated group reduced significantly post-irradiation (t =8.89-33.54,P ＜ 0.05),while the cell number of peripheral CD4 + CD25 + Tregs post-irradiation rose but not significantly.Thymic Treg cells increased 1 and 3 d post-irradiation(t =-6.45,-10.59,P ＜0.05),but reduced 28 d post-irradiation (t =5.34,P ＜ 0.05).Splenic Treg cells ascended obviously from 1 to 14 d post-irradiation (t =-6.82-3.89,P ＜ 0.05).After irradiation splenic Th17 cells increased at 1 d,and reached the maximal level at 3 d (t =-2.42,P ＜ 0.05),more obviously than splenic Treg cells.The reduction of Treg/Th17 ratio from 1 to 14 d post-irradiation disturbed Treg/Th17 balance and made it drift to the direction of Th17 (t =4.02-8.04,P ＜ 0.05).Conclusions Treg/Th17 imbalance plays an important role in immune injury induced by irradiation.

Objective To evaluate urinary nucleosides determination for the diagnosis of colorectal carcinoma. Methods The concentrations of 14 different urinary nucleosides from 42 colorectal carcinoma patients, 10 patients with villous adenoma and 62 normal adults were determined by high-performance liquid chromatography (HPLC); Principal component analysis was applied to classify colorectal cancer patients and normal adults. Results The levels of Pseu、 C、 m1A、 mU、 m22G、 I、 m1G、 ac4C、 m6A in colorectal carcinoma group were (42.67?21.89)、(0.44?0.51)、(2.72?0.83)、(0.13?0.08)、(1.79?0.55)、(0.50?0.36)、(1.44?0.52)、(0.82?0.30)、(0.07?0.08)nmol/(?mol creatinine) respectively, which were much higher than those in normal adults (P