1.The open-ended coaxial line method for colorectal cancer detection and depth evaluation of tumor invasion
Weiwei WANG ; Zhai CAI ; Shuai HAN ; Shiming LIN ; Miaoliang CHEN ; Zhou LI ; Song DUAN ; Xuegang XIN
The Journal of Practical Medicine 2016;32(20):3376-3379
Objective To confirm whether the open-ended coaxial line is effective in detection of the differences in dielectric properties between colorectal cancer tissues and surrounding normal tissues and evaluation of the depth of tumor invasion. Methods The open-ended coaxial line system at frequencies ranging from 50 to 500 MHz in 98 freshly excised colorectal cancerous specimens obtained from the operating theatre of Zhujiang Hospital, was used to detect both the relative permittivity and conductivity on the serosal surface of the carcinoma nidus, the mucosa of the carcinoma nidus, and the mucosa of the surgical resection margin. Pathological examinations were conducted on each specimen after surgery. Results The values for relative permittivity and conductivity of the colorectal cancerous mucosa were significantly higher than those of the normal mucosa (P < 0.01). For the tumor which had invaded or penetrated the serosa (stage ≥ T3), the dielectric properties of both the cancerous serosa and mucosa were higher than the one restricted to muscularis propria or less intestine wall (stage < T3) over the measured frequency range, and there existed statistical differences at the common frequencies of 213 MHz and 426 MHz. Conclusion The open-ended coaxial line system may result in fast and effective diagnostic differentiation between cancerous and normal colorectal tissues as well as reasonable assessment of the tumor infiltration depth.
2.Effect of short hairpin RNA on growth and proliferation of human bone marrow mesenchymal stem cells
Jie CHENG ; Zezhang TAO ; Honggang DUAN ; Bokui XIAO ; Shiming CHEN
Chinese Journal of Tissue Engineering Research 2008;12(16):3158-3162
BACKGROUND:Tetramethylpyrazine (TMP) can inhibit the expression of vascular endothelial growth factor (VEGF), but it is uncertain that TMP inhibit the growth and proliferation of HL-60 leukemic cells induced by VEGF.OBJECTIVE:To observe the effect of TMP on the proliferation of HL-60 leukemic cells induced by VEGF.DESIGN:Repetitive measurement and observation.SETTING:School of Medicine, Wuhan University of Science and Technology.MATERIALS:The experiment was carried out in the Molecular Biology Laboratory Center, School of Medicine, Wuhan University of Science and Technology from March to June in 2007. Human leukemic cell line HL-60 cells were purchased from Shanghai Institute of Cell Biology. TMP hydrochloride injection was produced by Wuxi Seventh Pharmaceutical Products Limited (Lot number:011014), protamine sulfate injection was produced by Shanghai First Biochemical Pharmaceuticals (Batch number:010302), and immunohistochemistry kit was purchased from Boster company.METHODS:①Human leukemic cell line HL-60 cells at log phase were used for the experiments. Cells were treated with 100 μg/L VEGF, and then TMP at final concentrations of 1.5, 15, 150 mg/L was added into culture medium. While the cells in medium without TMP were taken as blank control group, and the cells in medium with 20 mg/L protamine as positive control group. Meanwhile cells without treatment of VEGF were served as VEGF control group. After cells were incubated for 48 hours, the growth inhibiting rate of HL-60 cells was detected by MTT assay.②After HL-60 cells were treated with TMP at the final concentrations of 1.5, 15, 150 mg/L for 24 hours, the protein expression of VEGF in HL-60 cells was examined by SP immunohistochemistry.MAIN OUTCOME MEASURES:①Growth inhibiting rate of HL-60 cells.②Protein expression of VEGF.RESULTS:①Growth inhibiting rate of HL-60 cells:After HL-60 cells induced by VEGF were treated with 15 and 150 mg/L TMP, the absorbance value was significantly lower than that in VEGF control group (P < 0.05).②Protein expression of VEGF:After HL-60 cells were treated with TMP for 24 hours, the protein expression of VEGF was down-regulated with increasing TMP concentration in a dependent manner. Significant differences were observed in the protein expression of VEGF between cells treated by TMP and the controls (P < 0.01).CONCLUSION:shRNA, by targeting hTERT mRNA, has no noticeable influence on growth and proliferation of hMSCs, and might be safe for the somatic cells which are normal but do not express hTERT.
3.Antitumor activities of kushen flavonoids in vivo and in vitro
Mingyu SUN ; Jian ZUO ; Jifeng DUAN ; Jun HAN ; Shiming FAN ; Wei ZHANG ; Lifang ZHU ; Minghui YAO
Journal of Integrative Medicine 2008;6(1):51-9
OBJECTIVE: To explore the antitumor activities of kushen (Sophora flavescens) flavonoids (KS-Fs) in vivo and in vitro. METHODS: Cell proliferation was assayed by using methyl thiazolyl tetrazolium (MTT) method. H22 hepatocellular carcinoma and S180 sarcoma were induced in ICR mice. Lewis lung carcinoma was induced in C57BL/6 mice. H460 and Eca-109 tumor were induced in Balb/c nude mice by injecting 5x10(5) or 5x10(6) tumor cells in the right flank, respectively. RESULTS: KS-Fs could inhibit the growth of a variety of human tumor cell lines (A549, SPC-A-1, NCI-H460, etc.) in vitro. The antitumor efficacies were confirmed in the mice models of H22, S180 and Lewis lung tumors and the nude mice models of human H460 and Eca-109 xenografted tumors. The oral or intravenous maximum tolerated dose of KS-Fs was more than 2.8 g/kg or 750 mg/kg respectively, far more than the oral medial lethal dose of kushen alkaloids (< or = 1.18 g/kg). No adverse reactions were observed. CONCLUSION: These results suggest that KS-Fs or kurarinone may be developed as a novel antitumor agent.
4.Ketamine inhibits the excitability induced by formalin in spinal dorsal horn neurons of rats
Ming YAN ; Yinming ZENG ; Licai ZHANG ; Tijun DAI ; Shiming DUAN
Chinese Journal of Tissue Engineering Research 2006;10(46):195-197
BACKGROUND: It is indistinct that whether ketamine can exert antinociceptive effect througb influencing the transmission of nocuous information in spinal cord; Nitric oxide (NO) in spinal cord participates mainly in the formation and development of hyperalgesia, and it can also induce Fos protein expression. It is still controversal whether it contributes to the transmission and mediation of ketamine to pain signal.OBJECTIVE: To observe the response to formalin stimulation in spinal cord of the rats and the effect of ketamine.DESIGN: Balanced randomized animal trial.SETTING: Department of Anesthesiology, Affiliated Hospital of Xuzhou Medical College; Jiangsu Provincial Key Laboratory of Anesthesiology.MATERIALS: This trial was carried out in the Jiangsu Provincial Key Laboratory of Anesthesiology, Xuzhou Medical College from January to March 2000. Totally 30 Sprague-Dawley rats were chosen and balanced randomized into 6 groups: formalin group (n=6), formalin + ketamine group (n=6), ketamine +formalin group (n=6), ketamine group (n=6), formalin+normal saline group (n=3) and normal saline group (n=3). The gender ratio was the same in each group.METHODS: Formalin group:The rats were stimulated for one hour by subcutaneous injection of 0.05 volume fraction of 200 μL in the center of palm of unilateral fore-claw. Formalin +ketamine group: The rats were stimulated for 10 minutes by formalin, then for one hour by intraperitoneal injection of 100 rg/kg ketamine. Ketamine + formalin group: The rats were injected with ketamine for 10 minutes, then with formalin for one hour. Ketamine group: the same dosage of ketamine was intraperitoneally injected into the rats for one hour. Formalin + normal saline group: The rats were stimulated for 10 minutes by formalin, then intraperitoneally given 10 mL/kg normal saline for one hour. Normal saline group: the same volume of normal saline was intraperitoneally injected into the rats for one hour.MAIN OUTCOME MEASURES: ① Behavioral performance of the rats in each group. ② Spinal sections were chosen, and stained with c-fos genetic immunohistochemical and NADPH-d histochemical methods. The changes of the number of Fos-like immuno-positive neurons (FLI) and FLI/nitric oxide synthase (NOS) double-labeled neurons in the 4-layer sections (layer Ⅰ -Ⅱ ,layer Ⅲ-Ⅳ ,layerⅤ-Ⅵ ,layer Ⅶ-X )of spinal dorsal horn of the rats were observed.RESULTS: All the thirty rats entered the stage of result analysis. ① Behavioral changes: The rats of formalin group and formalin+ normal salinegroup had apparent pain response; Several minutes after injection with ketamine, righting reflex disappeared and did not recover at perfusion period.Prolonged sleep was found without obvious pain response performance. ② FLI neuron expression: A lot of FLI positive neurons were found in the spinal dorsal horn of injec tion side of the rats in the formalin group and formalin+ normal saline group, and they distributed principally in the layer Ⅰ - Ⅱ of spinal dorsal horn.The distribution in the ketamine + formalin group and formalin + ketamine group was basically similar to that in the formalin group and formalin + normal saline group, but positive neuron counts were significantly reduced (P < 0.01). ③ The expression of FLI/NOS double-labeled neurons: The number of double-labeled neurons in the spinal dorsal horn layer Ⅰ - Ⅱ of the rats in the ketamine+ formalin group and formalin+ ketamine group were significantly less than that in the formalin group and formalin+normal saline group [(1±1), (1±1), (7±3), (8±3),P < 0.01].CONCLUSION: Some neurons of ipsilateral corresponding spinal segments participate in the transmission and mediation of pain signal. Ketamine can suppress the activities of these neurons and exert antinociceptive effect. The antinococeptive function of ketamine may be caused by the activity depression of the NOS-positive neurons in spinal cord.
5.Influence of telomerase reverse transcriptase shRNA on expression of c-myc in nasopharyngea carcinoma cells
Yan WANG ; Shiming CHEN ; Zezhang TAO ; Bokui XIAO ; Honggang DUAN
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(07):-
OBJECTIVE To investigate the influence of short hairpin RNA targeting human telomerase reverse transcriptase(hTERT)on inhibition of telomerase activity and on expression of protein c-myc in nasopharyngeal carcinoma cells. METHODS Plasmid shRNA1 containing fluorescein gene and hTERT cDNA sequences were synthesized. Cells were transfected with plasmid shRNA1. The cell viability was examined using the MTT assay. The activity of telomerase was tested by polymerase chain reaction telomeric repeat amplification protocol- enzyme-linked immunosorbent assay (PCR-TRAP- ELISA),protein c-myc expression was tested by western blot. RESULTS It was observed that treatment with pshRNA1 in the presence of a valid transfection reagent could significantly reduce telomerase activity and the expression of protein of c-myc. CONCLUSION Inhibition of telomerase activity or expression of hTERT mRNA in nasopharyngeal carcinoma cells could inhibit cells proliferation and reduce the expression of protein of c-myc.
6.Study on anticonvulsant mechanism of ketamine
Jing MENG ; Yunpeng ZHAI ; Shiming DUAN ; Tao MA ; Tijun DAI
Chinese Pharmacological Bulletin 2003;0(09):-
Aim To observe the effects of ketamine on Na+,K+-ATPase,Ca2+-ATPase and NOSase activity in different cerebral cortex in convulsive mice.Methods The mice were randomly divided into blank group,normal saline(NS) group and ketamine 25 mg?kg-1 (KetⅠ),50 mg?kg-1(KetⅡ) group. The animals of blank group were killed directly.Convulsion was induced by intraperitoneally(ip) strychnine(1.5 mg?kg-1) in other groups,and correspond drugs were administered ip before five minutes.Action variety of mice was observed. Animals were killed on 30 minutes after strychnine injection.The activity of Na+,K+-ATPase,Ca2+-ATPase,TNOSase and iNOSase were assessed by spestrophotometric analysis in different cere-bral cortex(forehead,parietal and occipital area).Results Ketamine group could decrease mortality completely. The duration of tonic state in KetⅡ group was significantly shorter than that in KetⅠgroup.Compared with blank group,Na+,K+-ATPase and Ca2+-ATP ase activities were decreased in the group of NS and KetⅠ,and recovered normal level in the group of KetⅡ at parietal and occipital area. TNOS ase activity was decreased by 1/3 in KetⅡ group(P
7.Alfentanil protects the isolated rat heart against ischemia and reperfusion injury via opioid receptors and NO linked mechanisms
Meng CHEN ; Yinming ZENG ; Shiming DUAN ; Zhizhong SHEN ; Pengcheng XU
Chinese Journal of Clinical Pharmacology and Therapeutics 2000;0(02):-
AIM:?To?investigate?if?alfentanil?protects?the?isolated?rat?heart?against?myocardial?reperfusion?injury?and?if?the?mechanism?of?this?protection?is?mediated?via?opioid?receptors?and?NO-dependent?pathways. METHODS: Langendorff rat hearts were perfused at constant pressure with Kreb-Henseleit(K-H) buffer for 20 min?and?then?were?perfused?with?test?solution:?K-H?buffer?or?K-H?buffer?containing?alfentanil? 50 ?g?L -1,?alfentanil? 100 ?g?L -1, naloxone 200 ?g?L -1, alfentanil 100 ?g?L -1+naloxone 200 ?g?L -1, L-NAME 100 ?mol?L -1 and alfentanil 100 ?g?L -1+L-NAME 100 ?mol?L -1. After 10 min of this, the hearts were subjected to 25 min normothermic( 37 ℃) global ischemia followed by 30 min reperfusion with the same test solution as before. To evaluate myocardial function, LVEDP, LVDP, ?dp/dt max, HR and CF were measured at the 20th, 25 and 30th minute of perfusion and the 1st, 3rd, 5th, 10th, 20th and 30th minute of reperfusion. After experiment, the NOS and ATP content of myocardium were assessed. RESULTS: Before ischemia, alfentanil 100 ?g?L -1 decreased the HR at the 30th minute compared with the 20th minute(P
8.Protective effects of penehyclidine hydrochloride on transient forebrain ischemia reperfusion injury in gerbils
Tengfei MA ; Rong JIN ; Jing ZHANG ; Shuling GU ; Shiming DUAN
Chinese Journal of Clinical Pharmacology and Therapeutics 1999;0(04):-
AIM:To study the protective effects of penehyclidine hydrochloride(PHC) on transient forebrain ischemia reperfusion injury in gerbils.METHODS:The model of transient forebrain ischemia reperfusion was established in gerbils by bilateral carotid artery clamping.The effects of PHC on neurological function scores and the morphous of hippocampal pyramidal neuron of gerbils were observed after receiving transient forebrain ischemia reperfusion.SOD activities and contents of MDA in the hippocampus and cortex of gerbils were measured.RESULTS:In the groups of PHC(0.08),(0.24)(mg?kg~(-1)) and atropine,the stroke index was decreased,compared with the ischemia-reperfusion group after the gerbils received transient forebrain ischemia reperfusion for six hours.PHC could reduce the degree of injury in hippocampal pyramidal neuron after ischemia reperfusion for three days.CONCLUSION: PHC has protective effects on transient forebrain ischemia reperfusion injury in gerbils.
9.Effect of muscimol and bicuculline on brain cAMP content in rats undergoing isoflnrane anesthesia
Xingguo HU ; Shiming DUAN ; Yinming ZENG ; Jun WANG ;
Chinese Journal of Anesthesiology 1995;0(10):-
Objective To investigate the effects of GABAA receptor agonist-muscimol and GABAA receptor antagonist-bicuculline on brain cyclic adenosine monophosphate (cAMP) content in rats undergoing isoflurane anesthesia. Methods Forty-eight SD rats of both-sexes weighing 200-250g were randomly divided into 6 groups: group I received intraperitoneal (ip) normal saline (NS) 10ml.kg-1 (control group); group II in which animals inhaled 1.4% isoflurane for 30 min, 30 min after NS ip (isoflurane group);group IE in which animals inhaled 1.4% isoflurane for 30 min, 30 min after muscimol Img-kg ip (muscimol + isoflurane group); group IV in which animals inhaled 1.4% isoflurane for 30min, 30 min right after bicuculline 8 mg.kg-1 ip (bicuculline + isoflurane group); group V received muscimol lmg. kg-1 (muscimol group); group VI received bicuculline 8mg.kg (bicuculline group). The animals were decapitated 1h after 30min isoflurane inhalation or intraperitoneal NS or muscimol or 5min after bicuculline ip (rats developed convulsion within 7 min after bicuculline ip) . Brain was removed immediately for determination of cAMP content of cortex or brain stem. Loss of righting reflex was taken as sign of anesthesia. Brain cAMP content was measured by competitive protein binding assay. Results Muscimol shortened the time of loss of righting reflex induced by isoflurane (P
10.Effects of isoflurane on cyclic adenosine monophosphate content in brain of rats
Xingguo HU ; Shiming DUAN ; Yinming ZENG
Chinese Journal of Anesthesiology 1994;0(01):-
Objective To investigate the effects of isoflurane on cyclic adenosine monophosphate (cAMP ) content of brain in the rats. Methods Fourty SD rats were allocated randomly to 5 groups: no administration(control group,n=8), inhalation of 1.4% isoflurane until losing of righting reflex(loss of righting reflex group,n=8), inhalation of 1.4% isoflurane lasting 30 min(anesthesia group,n=8) ,righting reflex recovery after cessation of 30-min inhalation of 1.4% isoflurane (recoveryⅠ group,n=8) and 30 min after cessation of 30-min inhalation of 1.4% isoflurane (recovery Ⅱ group,n=8). The rats of each group were decapitated at the end of procedures to measure the cAMP content of brain tissue with competitive protein binding assay.Results As compared with that in control group,the cerebrocortical cAMP content only in anesthesia group significantly increased by 49% (P

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