1.Effect of IFN-γ inhalation on some cytokines of immunocompromised rats
Zhuang MA ; Guisheng QIAN ; Guijun HUANG ; Baolin MAO
Journal of Third Military Medical University 2001;23(2):-
Objective To study the effect of IFN-γ inhalation via aerosol on cytokines of the immunocompromised rats. Methods Immunocomprised rat model was established with cortisol acetate injection for 14 d and then Candida albicans fluid was injected by tracheal for establishing am immuno comprised with pulmonary infection model. IFN-γ was inhaled with aerosol 1 d before the bacterium injection and then for 1, 3 and 7 d respectively. The activity of TNF-α, and the levels of IL-1β and IL-6 in the supernatant of the cultured alveolar macrophage(AM), the activity of IFN-γ and TNF-α in bronchial alveolar lavage fluid (BALF), the expressions of IFN-γ,TNF-α, IL-1β, and IL-6 of the lung tissues, the level of IFN-γ,IL-1β, and IL-6 in the serum were investigated. Results The activity of TNF-α, and the levels of IL-1β and IL-6 in the culture supernatant of the AM of the rats treated with IFN-γ were significantly higher than those of the control. The activity of IFN-γ and TNF-α in BALF was higher in the IFN-γ inhaled rats than in the control (except the activity of TNF-α on the 7th day). The expressions of IFN-γ and IL-1β in lung tissues was higher in the rats treated with IFN-γ than in the control. The expression of TNF-α in the rats treated with IFN-γ was less than that in the control rats. The expression of IL-6 had no difference between 2 groups. And no difference was found in the activity of IFN-γ, and the levels of IL-1β and IL-6 in the serum between 2 groups(except IL-1β on the 3rd day). Conclusion Administration of IFN-γ via aerosol can obviously increase the activity or levels of some cytokines in the lung of the immunocompromised rats, but has no effect on them in serum of the immunocompromised rats.
2.Study on the safety of massive lung lavage to dogs after acute serious smoke inhalation
Fachuan NIE ; Zongcheng YANG ; Zhiyuan LIU ; Ya WU ; Qizhi LUO ; Yuesheng HUANG
Journal of Third Military Medical University 2001;23(3):327-
Objective To explore the feasibility and safety of massive lung lavage to dogs in the early stage after acute serious smoke inhalation. Methods A total of 16 dogs were inflicted with smoke inhaled injury and then divided into group A (n=5) without lung lavage, group B (n=6) in which left lung lavage was followed by lavage on the right 30 min later, and group C (n=5) in which the interval of the lavages on the left and right lung was 4 h. Massive lung lavage was carried out in 1 h after the injury under the guiding fiberoptic bronchoscopy and a special ventilation jet was served to supply enough oxygen to the dogs. The stability of pulmonary function and blood circulation were observed at different time points in every group. Results The average operating time of unilateral lung lavage was about 15 minuted and the liquid volume left in every lung after operation was from 100 ml to 140 ml when the total lavaging volume was kept in 250 ml. The changes of hemodynamic parameters were identical in Group B and Group C. Inhalation injury had certain negative effect while lung lavage almost had no effect on the blood circulation of the animals in 2 lavage groups. No serious cardiac arrythemia was found in them during the lavaging. Pulmonary function in Group C was better than that in Group B but worse than that in Group A. In the 2 lavaging groups, PaO2 remained higher than 9.33 kPa(FiO2=0.4)during and after the lavaging. Conclusion The ventilation mode and the lavage method introduced here ensure the safety of massive lung lavage in dogs with acute smoke serious inhalation injury.
3.Antishock effect of arginine vasopressin and its relationship with Rho kinase
Yuqiang FANG ; Tao LI ; Liangming LIU
Journal of Third Military Medical University 2003;0(13):-
Objective To investigate the relationship of antishock effect of arginine vasopressin(AVP) on hemorrhagic shock and its relationship with Rho kinase.Methods Hemorrhagic shock(40 mmHg for 2 h) Wistar rats were treated respectively by AVP(0.4 U/kg),Rho kinase specific inhibitor,Y-27632(30 ?g/kg),or these 2 reagents.The pressor effect of norepinephrine(NE) on these rats and the contractility of their isolated superior mesenteric artery(SMA) were observed.Isolated SMAs from hemorrhagic shock rats were adopted to observe the effects of AVP on vascular reactivity and calcium sensitivity and its relationship to Rho kinase with an isolated organ perfusion system.Results AVP at the concentration of 0.4 U/kg significantly improved the pressor effect of NE and the contractile response of SMA.While,Y-27632(30 ?g/kg) abolished these beneficial effects of AVP.The vascular reactivity and calcium sensitivity of SMA were significantly decreased following hemorrhagic shock.AVP at the concentration of 0.5 and 5 nmol/L significantly increased the decreased vascular reactivity and calcium sensitivity.These effects of AVP were abolished by Y-27632(10 ?mol/L).Conclusion Rho kinase may take part in the action of AVP on hemorrhagic shock via improving shock induced vascular hyporeactivity and calcium desensitization.
4.Effects of depression on collagen levels during wound healing in rats
Mingfang ZHOU ; Tianmin CHENG ; Zhengzhi FENG
Journal of Third Military Medical University 2003;0(13):-
Objective To measure the collagen content in the granulation tissue of skin wound surface in rats with depression in order to explore the possible mechanism about depression to wound healing.Methods Totally 126 Sprague-Dawley rats were randomly and equally allocated to 3 groups: simple wound group,depression plus wound group,wound plus depression group.Unpredictable stimuli,including fasting,electrocution,water deprivation,swimming in cold water and others,were performed for 18 d to establish depression model of rats.A full-thickness dermal wound(2.2 cm in diameter) was made on the back of rats to build wound model.Rats from depression plus wound group received 18 days' stimuli and then wound cutting.For those of wound plus depression,stimuli were carried out from the next day of wounding.The basal tissue in the center of the wound was taken out from 6 rats of every group in 3,5,7,10,14,18 and 21 d after wounding.Hydroxyproline reagent kit was used to detect the content of hydroxyproline in collected tissue.Wound collagen depositions in every time point were stained by nitroxanthic acid sirius red and then observed by polarization microscopy.Results Hydroxyproline content and collagen deposition in wound granulation tissue of depression plus wound group rats were lower than those in the other 2 groups.From 18 d after wounding,rats of depression plus wound group had a tendency to the same level as wound plus depression group.Conclusion Depression plus wound significantly decreases hydroxyproline synthesis,reduces collagen deposition in wound,and then delays the wound healing.
5.Experimental study on hemodynamics of traumatic brain injury using dynamic CT perfusion image
Journal of Third Military Medical University 2003;0(13):-
Objective To investigate the changes of region cerebral homodynamic and cerebral vascular autoregulation in traumatic brain injury(TBI) by dynamic CT perfusion(CTP) imaging in rat.Methods Forty-five SD rats underwent CTP,including 10 control rats and 35 TBI rats,on 1,6,24 h,and day 3,7,14,21 after TBI.The region of interest(ROI) was located near traumatic area within 2 mm and mirror area of relative brain hemisphere.False color pictures and dynamic rCBF,rCBV,MTT and PS were obtained in different time.Pathological examination was also done.Results The rCBF,rCBV on 1 h after TBI was lower than that of control group,and significantly higher on 6-24 h after TBI.They reached the peak on day 3.PS had a similar change compared with rCBF,rCBV,but a longer duration.MTT showed to moderate changes except an isolated tide on 3 days after TBI.The rCBF on mirror area of relative brain hemisphere was almost stable except 3 days after TBI,which was lower than that of control group.Meanwhile,the rCBV on mirror area of relative brain hemisphere was normal.Conclusion CTP is an advanced technique to investigate the changes of region cerebral homodynamic and cerebral vascular autoregulation in TBI.The early hypoperfusion and next hyperperfusion around traumatic areas after TBI suggest that the dysfunction of cerebral vascular autoregulation should be committed to the change of homodynamic in TBI.
6.Glucocorticosteroid membrane implanted into perichoroidal space protects retina of rabbit endophthalmitis
Journal of Third Military Medical University 2003;0(13):-
Objective To evaluate the effect of implantation of chitosan membrane with triamcinolone(TA) into perichoroidal space combined with intraocular vancomycin injection on the protection of retina in rabbit endophthalmitis.Methods The drug membrane was made of gelatin-chitosan.The right eyes of 30 normal pigment rabbits were inoculated intravitreally with 0.1 ml mixture of 105 CFU standard strain Staphylococcus aureus(ATCC 25923).Twenty-four hours later,rabbits were equally randomized to intravitreal vancomycin group(Group A),intravitreal vancomycin and TA eye dropping(Group B),and intravitreal vancomycin combined with TA membrane implantation(Group C).Clinical observation of ocular fundus was done every day with indirect ophthalmofundoscope.B ultrasound examination was carried out in 24 h after inoculation and on the 14th day after therapy.Electroretinogram(ERG) was done before therapy and the 14th day after therapy.Histopathologic grading and electron microscopy were carried out at the 14th day.Results After implanting the drug membrane into perichoroidal space,the inflammation in group C was markedly less than the other 2 groups,and its inflammation score was significantly lowest(P
7.Lipoteichoic acid of Bifidobacterium down-regulates VEGF expressions and inhibits the metastasis of colon carcinoma
Yanhua PENG ; Jianlong LIU ; Yue WANG ; Mingfang LIU ; Shuhui CHEN
Journal of Third Military Medical University 2003;0(13):-
Objective To study the effect of lipoteichoic acid(LTA) of Bifidobacterium on the expression of vascular endothelial growth factor(VEGF) and invasion and metastasis of colon carcinoma cell lines.Methods The effects of LTA on adhesion,invasion and metastasis of human colon carcinoma lines LoVo and HT-29 in vitro were examined by MTT colorimetric and transwell chamber.The mRNA and protein expressions of VEGF in the two colon carcinoma cells treated with LTA were detected by RT-PCR and Western blot.Results The adhesion ability decreased in LoVo cells and HT-29 cells after the treatment of LTA(20,50,80 ?g/ml) for 30,60,90,120 min(P
8.Effect of CO_2 pneumoperitoneum on expressions of ?_1-integrin and E-cadherins in endometrial cancer cells transplanted into nude mouse
Shi CHEN ; Zhiqing LIANG ; Junling YAN
Journal of Third Military Medical University 2003;0(13):-
Objective To discuss the effect of CO2 artificial pneumoperitoneum on the invasion of the endometrial cancer in nude mouse resulted from the transplantation of the cancer cells and its mechanism.Methods Thirty nude mice were divided into 3 groups based on the time in the CO2 pneumoperitoneum circumstance.Control group: the small intestine of the nude mouse was exposed in air for 5 min,and the cancer cells were injected into right lower quadrant after suture.The gas was depleted after 40 min.40 min group: CO2 gas was poured into abdominal cavity to form a 4 mmHg artificial pneumoperitoneum for 5 min before cancer cells were injected into right lower quadrant.The gas were depleted after 40 min.80 min group: CO2 gas was poured into abdominal cavity to form a 4 mmHg artificial pneumoperitoneum for 5 min before cancer cells were injected into right lower quadrant.The gas were depleted after 80 min.The time that each group took to form a solid tumor was recorded.Four weeks later,the transplantation tumors were taken out and sliced into frozen sections and paraffin-embedded sections.The expression of ?1-intergrin and E-cadherins was detected by IMF.Results The time taken to form the solid cancer was shorter in the 40 min group and 80 min group than in the control group,with more blood vessel found(P0.05).Conclusion The CO2 pneumoperitoneum could enhance the abilities of invasion and adhesion of endometrial cancer cells,which is associated with the expression changes of ?1-intergrin and E-cadherins in the cancer cells.
9.Dexamethasone inhibits cervical carcinoma cell xenografted tumors in nude mice
Jianhua ZENG ; Ling ZHONG ; Bo XU
Journal of Third Military Medical University 2003;0(13):-
Objective To investigate the effects of over physiological dose of dexamethasone on cervical carcinoma cell xenografted tumors in nude mouse.Methods HeLa cells were inoculated subcutaneously in BALB/c female mice aged 5-6 weeks.Over physiological dose of dexamethasone(62.5 mg?kg-1?d-1) was injected into the peritoneum from the 16th day after inoculation for 10 d.Diameters of tumor were measured every two or three days.The volumes and the inhibition rates of tumor were calculated.Apoptotic bodies were observed by transmission electron microscopy and counted by TUNEL.VEGF and TNF-? were detected by immunohistochemistry.Results In vivo,over physiological dose of dexamethasone can repress the growth of tumor and induce cell apoptosis.Compared with the control group,the tumor sizes were significantly smaller(P0.05).Conclusion Over physiological dose of dexamethasone can repress cervical tumor growth in vivo.This action may be related to apoptosis and VEGF,but have no relationship with TNF-?.
10.Construction of Skp2 shRNA and its effect on lung cancer cell growth
Sheng LI ; Tonghua MEI ; Lian LI ; Mingchuan ZHANG
Journal of Third Military Medical University 2003;0(13):-
Objective To construct and identify the RNAi eukaryotic vector of Skp2 gene and to observe its interfering effect on the growth of SPC-A-1 lung cancer cells.Methods The specific shRNA sequence was designed and synthesized according to the Skp2 cDNA sequence in GenBank.The sequence was cloned into plasmid pGenesil-1.Then recombinant vector was transfected into SPC-A-1 lung cancer cells by Lipofectamine 2000.The expressions of Skp2 mRNA were analyzed by RT-PCR and the levels of Skp2 protein were detected by Western blot.The cell growth suppression was analyzed by MTT assay.Distribution of cell cycle was assessed by flow cytometry.Results The sequence of template and specific siRNA was correct by sequence analysis.Obvious decrease was observed in the levels of Skp2 mRNA and Skp2 protein after Skp2 shRNA transfection(P