2.Gene therapy of human ovarian carcinoma in vitro
Pu LI ; Dongtian LI ; Bingnan YIN ; Hua TANG ; Lirong YIN
Chinese Journal of Clinical Oncology 2014;(13):861-865
To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.
3.Experimental Study on Effect of Arsenic Trioxide on Suppression of Neuroblastoma Cell Invasion
hua, YIN ; suo-qin, TANG ; chen, FENG ; fang, YU
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate whether arsenic trioxide(As2O3)with different density is capable of affecting the invasiveness of neuroblastoma(NB)cells,and to give grounds for NB therapy with As2O3.Methods 1.Well-developed NB cells were selected and exposed to 0.75 ?mol/L,1.50 ?mol/L,3.0 ?mol/L As2O3 for 24 h;2.Collect the adherence cells,count the number and float them in nutrient medium again,add them into the transwell polycarbonate membrane plate that was covered by matrigel,there were 2?104 NB cells in each well;3.After 24 h,take off the membrane,fix the cells which cross the membrane with mehanol and dye them with hematoxylin;4.Observe the NB cells and count them,so the capability of invasion of LA-N-5 was evaluated by transwell chamber assay.Results After 24 h with the different density As2O3,the number of invading LA-N-5 cells was significantly lower in As2O3 group than that in control group(the number of invading cells of the As2O3 group was 28.0?4.0,19.33?4.16,6.33?1.53,respectively,the cell number of the control group was 46.33?6.11)(P=0.013,0.003,0);among the experiment groups,there was no difference between 0.75 ?mol/L and 1.50 ?mol/L(P=0.06),and it was significantly different between 0.75 ?mol/L and 3.0 ?mol/L,1.50 ?mol/L and 3.0 ?mol/L(P=0,0.007),the number of invading LA-N-5 cells of 3.0 ?mol/L As2O3 was the least.Conclusions As2O3 could inhibit the invasive potential of NB cells;the inhibitory action of 3.0 ?mol/L As2O3 is the most.
4.Effect of medicines for activating blood and reinforcing Qi on angiogenesis in infarcted myocardium edge area of acute myocardial infarction model in rats.
Wen-Hua ZANG ; Shen-Hua YIN ; De-Cai TANG ; Bing-Bing LI
China Journal of Chinese Materia Medica 2014;39(5):901-906
OBJECTIVETo study the effect of medicines for activating blood and reinforcing Qi on the number of new micro-vessels and the protein expressions of VEGF and bFGF in the infarcted myocardium edge area of acute myocardial infarction (AMI) model in rats.
METHODThe AMI model of rats was established. After the successful model establishment, rats were randomly divided into the sham-operated group, the model group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group, the Chuanxiong-Huangqi (1 : 2) group, the Danshen group, the Chuanxiong group, the Chishao group and the Shexiang Baoxin pill group, with five rats in each group. Rats in each medicated group were orally administered with drugs as per 13.5 g x kg(-1) x d(-1) once everyday for three weeks. The immunohistochemical SP method was adopted to detect the expression of vWF in myocardial tissues, and count the number of micro-vessels (MVC). The protein expression of VEGF and bFGF in myocardial tissues were determined by Western blot.
RESULTThe new micro-vessels stained by vWF factor could be found in the infarcted myocardium edge area of the sham-operated group, the model group and all of medicated groups. The sham-operated group show unobvious new micro-vessels in myocardial tissues. A small amount of new micro-vessels could be seen in the infarcted myocardium edge area of the model group. Whereas a larger number of micro-vessels could be seen in the infarcted myocardium edge area of all of medicated groups. The differences between the sham-operated group and the model group had statistical significance (P < 0.05). The differences between each medicated group and the model group had statistical significance as well (P < 0.05 or P < 0.01). The lowest protein expression of VEGF and bFGF was found in myocardium of the sham-operated group, with the statistical significance compared with the model group (P < 0.05). Compared with the model group, each medicated group showed significant increase in the protein expression of VEGF and bFGF, with the statistical significance between them (P < 0.05 or P < 0.01).
CONCLUSIONThe Danshen group, the Chuanxiong group, the Chishao group, the Danshen-Huangqi (1 : 2) group, the Danshen-Huangqi (1 : 1) group and the Chuanxiong-Huangqi (1 : 2) group show the effect in promoting angiogenesis. Their mechanism for promoting angiogenesis may be related to the improvement of the protein expressions of VEGF and bFGF, so as to increase the contents of VEGF and bFGF and promote the angiogenesis of new vessels.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Humans ; Male ; Microcirculation ; drug effects ; Microvessels ; drug effects ; physiopathology ; Myocardial Infarction ; drug therapy ; physiopathology ; Neovascularization, Pathologic ; drug therapy ; genetics ; metabolism ; Qi ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism
6.Effects of different acupuncture and moxibustion methods on ultrastructure of gastrocnemius in rats.
Ming GAO ; Hua-Yuan YANG ; Tang-Yi LIU ; Xun-Jie GU ; Yin-E HU ; Gang XU ; Wen-Chao TANG
Chinese Acupuncture & Moxibustion 2014;34(3):261-265
OBJECTIVETo explore the efficacy differences in early intervention of different acupuncture and moxibustion methods on gastrocnemius fatigue in rats induced by electrical stimulation.
METHODSFifty male SD rats were randomly divided into a control group, a model group, a hand acupuncture group, an electroacupuncture group and a moxibustion group, 10 rats in each group. Electrical stimulation of the sciatic nerve was given in the control group and gastrocnemius fatigue test was induced by electrical stimulation of the sciatic nerve in the model group after anesthesia without other treatment, but just take 6 times interval stimulation in the control group. The hand acupuncture group, the electroacupuncture group and the moxibustion group were treated with the corresponding acupoints stimulation method respectively for 20 min before gastrocnemius fatigue test, and Dazhui (GV 14) and Zusanli (ST 36) were selected. Immediately after gastrocnemius fatigue test, three or four gastrocnemius tissues at the same site on the right side were quickly taken for making specimen for transmission electron microscope (TEM). The changes of skeletal muscle ultrastructure of myofibrils, mitochondria, sarcoplasmic reticulum, glycogen particles were observed under TEM.
RESULTS(1) Muscle fibers disorder, partial mitochondrial vacuolization and glycogen particles smaller were shown in the model group. (2) No abnormalities were shown in the hand acupuncture group and the moxibustion group with mitochondrial morphology and number, which better than that in the model group, and glycogen particles increased. (3) Abnormal changes in morphology were shown in the electroacupuncture group with part of the muscle fibers derangement, Z line malalignment and a few mitochondria vacuolization.
CONCLUSIONHand acupuncture, electroacupuncture and moxibustion have the different effects on ultrastructure of gastrocoemius in rats. Acupuncture and moxibustion have shown good effects on the prevention and treatment of exercise-induced skeletal muscle cell and organelle damage and delaying exercise-induced fatigue.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Glycogen ; metabolism ; Male ; Mitochondria ; metabolism ; ultrastructure ; Moxibustion ; Muscle, Skeletal ; metabolism ; ultrastructure ; Rats ; Rats, Sprague-Dawley
7.Electroacupuncture therapy and corrosion of acupuncture needle.
Yan-Hong SUN ; Tang-Yi LIU ; Hua-Yuan YANG ; Ming GAO ; Yin-E HU ; Gang XU ; Wen-Chao TANG
Chinese Acupuncture & Moxibustion 2014;34(12):1238-1240
During the process of electroacupuncture (EA) therapy, whether there being a corrosive effect in ac- upuncture needles was observed. Acupuncture needles were inserted into a rabbit's acupoint to perform a 12-hour electrical stimulation with three types of common EA waveform; additionally two needles were put in 0.9% sodium chloride solution with 12-hour direct current. Afterwards, environmental scanning electron microscope was applied to detect the surface physical characteristics of acupuncture needles. As a result, after a 12-hour continued electri- cal stimulation with three types of common EA waveform in the rabbit, there was no corrosive effect in acupunc- ture needles; but the direct current could cause severe corrosion in acupuncture needles. It is believed that there is no corrosion effect on acupuncture needles in current EA treatment, and some accidents reported in literature may be related to quality of EA device or improper manipulation during the treatment.
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8.Epidemiology and risk factors for community-acquired blood stream infection caused by extended spectrum β-lactamases-producing Escherichia coli and Klebsiella pneumonia strains
Min ZHONG ; Kai ZHANG ; Xiangning HUANG ; Lin YIN ; Xin LIU ; Hua YU ; Wenfang HUANG ; Rongzhen TANG ; Ting FENG
Chinese Journal of Microbiology and Immunology 2016;36(2):117-123
Objective To investigate the incidences, risk factors, genotypes and epidemiology of community-acquired blood stream infection caused by extended spectrum β-lactamases (ESBLs)-producing Escherichia coli and Klebsiella pneumonia strains and to analyze the sensitivity of those ESBLs producing strains to commonly used antibiotics. Methods Forty-two patients who were diagnosed with community-ac-quired blood stream infection caused by Escherichia coli or Klebsiella pneumonia strains in Sichuan Provincial People′s Hospital were recruited in this study. Disc diffusion method was used for the phenotypic confirmato-ry test of ESBLs. Agar dilution method was performed to measure the antimicrobial susceptibility of the ESBLs-producing strains to 13 clinically commonly used antibiotics. Genotypes of the ESBLs-producing strains were identified by polymerase chain reaction (PCR). Multilocus sequence typing (MLST) was used to analyze the epidemiology of ESBLs-producing strains. Logistic regression analysis was performed to analyze the risk factors for community-acquired blood stream infection. Results The ESBLs-producing Escherichia coli strains accounted for 56. 3% (18 / 32) and the ESBLs-producing Klebsiella pneumoniae strains accounted for 20% (2 / 10). All of the 20 ESBLs-producing strains were sensitive to imipenem, meropenem, ertapen-em, nitrofurantoin and moxalactam. The ESBLs-producing strains sensitive to amikacin, piperacillin-tazobactam and fosfomycin accounted for 95% , 90% and 85% , respectively. Drug resistance rates of the 20 strains to cefotaxime, levofloxacin, ciprofloxacin and cefepime were relatively high accounting for 100% , 80% , 80% and 75% , respectively. Among the 20 ESBLs-producing strains, 7 strains only carried the CTM gene, while the other 13 strains were all positive for two genotypes of ESBLs, mainly identified as TEM+CTM-M-14 and TEM+CTM-15 genotypes. The 18 Escherichia coli strains were classified into 10 ST types, most of which were ST131 type, followed by ST10 and ST38 types. This study indicated that malignant tumor might be a possible risk factor. Conclusion The prevalence of community-acquired blood stream infection caused by ESBLs-producing Escherichia coli strains was becoming increasingly serious. Malignant tumor might be the risk factor associated with the producing of ESBLs in Escherichia coli and Klebsiella pneumonia strains. TEM+CTX-M-14 was the predominant genotype of ESBLs-producing strains and the prevalent clone was ST131 type. Carbapenems and enzyme inhibitor compounds were ideal drugs for the treatment of commu-nity-acquired blood stream infection caused by ESBLs-producing Escherichia coli and Klebsiella pneumonia strains. This study was limited by the small sample size. Therefore, it is necessary to conduct further resear-ches based on a large number of samples.
9.Inhibitory effect of scopolamine on withdrawal syndromes in morphine dependent rats
Qiang FU ; Xin-Hua WANG ; Xue-Yin SHI ; Yang LU ; Jing-Lan TANG
Academic Journal of Second Military Medical University 2001;22(4):351-353
Objective: To observe the inhibitory effect of scopolamine(Spm) and chlopromazine (Clo) on withdrawal syndromes in morphine dependent rats. Methods: The intensity of withdrawal syndromes on the model of morphine dependent rats was recorded after single or muiltiple subcutaneous administration(sc) of Spm and Clo at different doses. Results: Withdrawal syndromes were markedly decreased when single Spm 1 mg/kg and Clo 0.5 mg/kg combined with morphine were injected (P<0.05). Spm+Clo(sc) had much stronger effects on inhibiting withdrawal syndromes after intraperitoneal (ip) naloxone in morphine dependent rats (P<0.01). Conclusion: Spm can act on Ach-receptor and relieve morphine withdrawal syndromes. Clo may have a synergistic action with Spm via α2-receptor in the locus coeruleus of the rat brain stem.
10.Inhibitory effect of scopolamine on withdrawal syndromes in morphine dependent rats
Qiang FU ; Xin-Hua WANG ; Xue-Yin SHI ; Yang LU ; Jing-Lan TANG
Academic Journal of Second Military Medical University 2001;22(4):351-353
Objective: To observe the inhibitory effect of scopolamine(Spm) and chlopromazine (Clo) on withdrawal syndromes in morphine dependent rats. Methods: The intensity of withdrawal syndromes on the model of morphine dependent rats was recorded after single or muiltiple subcutaneous administration(sc) of Spm and Clo at different doses. Results: Withdrawal syndromes were markedly decreased when single Spm 1 mg/kg and Clo 0.5 mg/kg combined with morphine were injected (P<0.05). Spm+Clo(sc) had much stronger effects on inhibiting withdrawal syndromes after intraperitoneal (ip) naloxone in morphine dependent rats (P<0.01). Conclusion: Spm can act on Ach-receptor and relieve morphine withdrawal syndromes. Clo may have a synergistic action with Spm via α2-receptor in the locus coeruleus of the rat brain stem.