1.Establishment and assessment of a rat model of persistent intestinal mucosal injury induced by methotrexate
Du TU ; Zhihong XU ; Mingwei ZHANG ; Ruifen ZHANG ; Yuanyuan DENG
Acta Laboratorium Animalis Scientia Sinica 2015;(4):382-388
Objective To establish a rat model of persistent intestinal mucosal injury.Methods The rat model of persistent intestinal mucosal injury was induced by intraperitoneal injection of methotrexate.The food intake and body weight of all the rats were recorded.Pathological changes were observed using HE staining, the level of D-lactate and diam-ine oxidase in plasma, and myeloperoxidase and malondiadehyde in the intestinal tissue were measured by biochemistry. Results After modeling, the rat body weight and food intake were decreased.On day 4, the scores of mucosal damage, the levels of plasma D-lactate, DAO, MPO and MDA were significantly increased (P<0.05).On day 5, the intestinal damages of rats began to restore, and there was no significant difference among groups on day 6.The symptoms after the secondary injection were similar to those after the first injection, and the rats recovered gradually at day 12.Conclusions Intestinal mucosal injury in rats induced by 20 mg/kg MTX is an acute injury process, the course only lasts for 4-5 days. Intermittent injections twice of 10 mg/kg MTX can cause persistent intestinal mucosal injury in rats.This persistent injury model is more suitable for nutritional therapy evaluation in medium-and long-term studies of nutritional therapy.
2.Effects of peptide-based enteral nutrition on inflammatory response and immune function in intestinal mucositis rat
Du TU ; Zhihong XU ; Yuanyuan DENG ; Mingwei ZHANG ; Ruifen ZHANG
Chinese Journal of Clinical Nutrition 2015;23(1):41-47
Objective To evaluate the effects of peptide-based enteral nutrition (PBEN) on inflammatory response and immune function in rats with intestinal mucositis.Methods 48 Sprague-Dawley male rats were randomly divided into 6 groups (all n =8):basal feed group (BFG),PBEN group (PBENG),intact protein enteral nutrition group (IPENG),methotrexate (MTX) + BFG,MTX + PBENG,and MTX + IPENG.The rats in MTX + BFG,MTX + PENG,and MTX + IPENG were intraperitoneal injected with MTX 10 mg/kg on day 0 and day 6 to induce sustained intestinal injury.From day 1,BFG and MTX + BFG were fed with basal feed,PBENG and MTX + PBENG with PBEN,IPENG and MTX + IPENG with IPEN.The daily energy intake of each rat was 1.80 kJ/g body weight.All the rats were sacrificed on day 11.The pathological changes of intestinal tissue were observed with HE staining,the levels of tissue myeloperoxidase (MPO),nitric oxide (NO),inducible nitric oxide synthase (iNOS),the thymus index and spleen gland index of intestinal tissue were measured using colorimetry,and the serum levels of immunoglobulins IgG,IgA,and IgM were determined with enzyme-linked immunosorbent assay.Results There were no significant differences among BFG,PBENG,and IPENG in each index.Serious injury of intestinal mucosa was observed in MTX groups.Significant differences were noted in all indexes between MTX + BFG and BFG (all P <0.05).The mucosal damage score (Chiu score) and the level of MPO and iNOS in MTX + PBENG were significantly lower than those in MTX + BFG [2.3 ± 0.69vs.2.96 ± 0.75,P =0.003 ; (2.30 ± 0.42) U/g tissue vs.(2.98 ± 0.23) U/g tissue,P =0.040 ; (0.37 ±0.06) U/mg prot vs.(0.44 ±0.10) U/rag prot,P =0.030] ; the serum levels of IgG and IgA were significantly higher than those in MTX + BFG (P =0.015,P =0.021) ; however,the levels of NO and IgM were not significantly different between the 2 groups (P =0.597,P =0.160).There were no statistically significant differences between MTX + IPENG and MTX + BFG in terms of the indexes (all P > 0.05).Conclusion PBEN can reduce the inflammation response and improve the immune function in intestinal mucositis rat.
3.Study on the Sustained Release of BSA from Chitosan-OREC/BSA Films Coated Mats in Vitro
Ruifen XU ; Qingxia ZHAO ; Xin CHEN ; Xingxing ZHENG ; Xuyang FENG
Journal of Modern Laboratory Medicine 2017;32(4):119-121,124
Objective To investigate the sustained release of BSA from chitosan-OREC/BSA films coated mats in vitro.Methods The negatively charged cellulose acetate (CA) fibrous mats were modified with multilayers of the positively charged chitosan or chitosan-OREC intercalated composites and the negatively charged bovine serum albumin (BSA) via electrostatic layer-by-layer (LBL) self-assembly technique.The adsorption and rinsing steps were repeated until the desired number of deposition bilayers was obtained.The in vitro BSA encapsulation and release experiments demonstrated that OREC could affect the degree of protein loading capacity and release ficiency of the LBL films coating.Results In the pH-gradient release assay,only a small amount of BSA was released from the mats in 1 h.As the time increased,the release rate of BSA of all the samples gradually went up to the maximum data within 8 h.For the samples with identical number of bilayers and record time,obvious increasing of the release amount could be seen in pH 7.4,in comparison with pH 1.2.Besides,doubling bilayers film-coated mats generally.Meanwhile,it was slightly distinguishable between 5 and 5.5 as well as 10 and 10.5 bilayers (t=0.651~ 1.324,P>0.05).Interestingly,it could be seen that protein release of the chitosan-OREC/BSA films coated mats remarkably increased compared with that of chitosan/BSA films coated mats(t=2.264~ 2.305,P<0.05).Conclusion The release of protein in the initial time could be controlled by adjusting the number of deposition bilayers,the outmost layer and the composition of coating bilayers.
4.Effect of constant magnetic fields on intracellular free calcium of human umbilical arterial vascular smooth muscle cells
Xuyang FENG ; Ruifen XU ; Bo SONG ; Haichang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1982;0(01):-
Objective To study effects of constant magnetic fields(CMF) on the intracellular free calcium of human umbilical arterial vascular smooth muscle cells to discuss the mechanism of CMF affecting the proliferation of VSMC.Methods The intracellular free calcium concentration of human umbilical arterial vascular smooth muscle cells was detected by laser scanning confocal microsopy.Results The CMF affected the intracellular free calcium concentration of human umbilical arterial vascular smooth muscle cells.The CMF(5 mT) caused a persistent decrease of intracellular free calcium concentration of VSMC from 10 min to 50 min.Conclusion The CMF could inhibit proliferation of VSMC by decreasing the intracellular free calcium constant magnetic field concentration of VSMC.The CMF could markedly inhibit proliferation of VSMC,which might be useful in treatment of RS after PTCA.
5.Preparation of pellets in Weigukang Capsule
Guizhi SUN ; Zhihua CAO ; Wen LIU ; Ruifen JU ; Shaoxi XU
Chinese Traditional Patent Medicine 1992;0(04):-
AIM: To investigate the preparation and parameter of pellets of traditional Chinese medicine in order to solve the problem of preparation of pellets of traditional Chinese medicine. METHODS: We took centrifugal spray method to prepare the pellets, and compared effect of spray coating and fluidzed bed coating on the quality of pellets. RESULTS: The best preparation of pellets was determined as followed. The frequency of turntable was 45 HZ, the flow rate of liquid was 1.2 g/min. the relative density of liquid was 1.20 g/cm 3; when coating weigh reached 14%, the better pellets could be obtained. CONCLUSION: The need of the assistant matter of taking spray pellets and film coating was less and roboticized. It accorded with the answer of GMP and the regular production of the preparation of pellets of traditional Chinese medicine.
6.Low dosage sufentanil for blind nasotracheal intubation
Yuan LI ; Lixian XU ; Guoliang ZHANG ; Lingling LU ; Hui ZHANG ; Ruifen XU
Journal of Practical Stomatology 2000;0(06):-
Objective:To evaluate the effects of low dosage sufentanil used for blind nasotracheal intubation.Methods:Sixty cases for maxillofacial surgery were divided into 3 groups randomly with 20 in each group. Patients in groupⅠwere administered with fentanyl and midazolam by vein, those in groupⅡ with fentanyl and droperidol by vein,those in group Ⅲ with sufentanil at 0.1~0.2 ?g/kg. Intramuscular premedication of atropine-midazolam and blind nasotracheal intubation were performed in all cases after surface anesthesia for routine. Blood pressure(BP), heart rate (HR), mean artery pressure (MAP), blood oxygen saturation of pulse (SpO_2), intubation complication(IC), intubation achievement ratio(IAR), sedation score (Ramsay score), patient satisfaction(PS), and the incident rate of amnesia(IRA) in the three groups at T1 (before administering drug), T2 (after administering drug) and T3 (when tracheal tube was inserted into tracheal) were measured and observed. Results:In groups I and Ⅱ Ramsay score increased to 3~5, MAP and SpO_2 decreased (P0.05).Conclution:The low dose sufentanil can be applied for the sedation and analgesic before blind nasotracheal intubation.
7.Myocardial damage and changes of plasma and myocardial angiotensin after craniocerebral injury in rats
Caixia GUO ; Fenghe DU ; Like ZHANG ; Xiuying XU ; Ruifen CHEN ; Yilin SUN ; Shida HE ; Shumin WANG
Chinese Journal of Tissue Engineering Research 2005;9(41):152-154
BACKGROUND: Craniocerebral injury can cause a series of visceral complications, among which cardiovascular complication is paid special attention.OBJECTIVE: To investigate the effects of craniocerebral injury on changes of circulatory and local angiotensin Ⅱ (Ang Ⅱ ) and local angiotensin Ⅱ receptor 1 (AT1) in the heart.DESIGN: Randomized controlled experiment taking animals as subjects.SETTING: Beijing Tiantan Hospital, and the College of Basic Medicine,Capital University of Medical Sciences.MATERIALS: The experiment was conducted at the Central Laboratory of Capital University of Medical Sciences and the Central Laboratory of Beijing Tiantan Hospital from 2003 to 2004. Totally 40 healthy male Wistar rats were divided randomly into craniocerebral injury group and control group with 20 in each group.METHODS: Rats in craniocerebral injury group were treated with weightdrop method to establish the model of craniocerebral injury, while rats in control group received no impact. Twenty-four hours after hitting, 10 rats in each group were selected to assay their Ang Ⅱ and AT1; the other 10 in each group were selected to observe their myocardial forms.myocardium of rats assayed with light microscope after hematoxylin-eosin staining and transmission electron microscope.It was significantly higher in craniocerebral injury group than in control ity: It was obviously higher in craniocerebral injury group than in control Ⅱ and AT1: The area of positive reactant and gray value in craniocerebral toxylin-eosin staining: Strong acidophil staining was found on myocardial cellular plasma in craniocerebral injury group. The results showed that cytoplasm shrank obviously; muscle fiber broke, decreased or disappeared.Focal hydropic degeneration, lysis or necrosis was observed in myocardium.Ultrastructural pathological observation revealed pathological damage of myocardium.CONCLUSION: Craniocerebral injury in rats can cause myocardial damage, and changes of angiotensin system may be one of the factors.
8.Natural anti-oxLDL IgM monoclonal antibody in the pathogenesis of atherosclerosis
Xuyang FENG ; Ruifen XU ; Yan GAO ; Haokao GAO ; Zheng HE ; Haichang WANG
Journal of Geriatric Cardiology 2009;6(1):42-48
Objective To explore the role and the possible molecular mechanisms of natural anti-oxLDL IgM monoclonal antibody played and involved in pathogenesis of atherosclerosis. Methods Natural anti-oxLDL IgM monoclonal antibody 3A6 was generated by using standard hybridoma production techniques. Influence of 3A6 on formation of foam cells was observed by Oil Red O staining and affinity of Na125I-conjugated oxLDL on the naive and LPS-activated macrophages. After LPS stimulation on macrophages, anti-TLR4 neutralizing mAb, p38MAPK specific inhibitor SB203580, NF-kB specific inhibitor PDTC or RNAi targeting Fcα/μ receptor (Fcamr) were applied, respectively. Results Natural anti-oxLDL IgM monoclonal antibody 3 A6 were found specifically inhibit the binding of CuoxLDL to naive macrophages but not the binding of CuoxLDL to LPS-activated macrophages. It also promoted the formation of CuoxLDL-mediated foam macrophages. 3A6 F(ab')2 or pre-incubation with un-related IgM inhibited the binding of 3A6/CuoxLDL complex to LPS-activated macrophages. LPS up-regulated the expression of Fcamr in macrophages in a dose- and time-dependent manner, which was attenuated by treatment with anti-TLR4. LPS induced the phosphorylation of p38MAPK and translocation of NF-kB p65, contributing to the up-regulated expression of Fcα/μ receptor in macrophages. Conclusions Natural anti-oxLDL IgM monoclonal antibody 3A6 specifically inhibited the binding of CuoxLDL to naive macrophages in vitro. However, LPS, through the Toll-like receptor (TLR)4 receptor, activated the p38MAPK and NF-kB pathways and up-regulated the expression of Fcα/μ receptor in macrophages, which promoted the binding of 3A6/CuoxLDL complex to macrophages through binding with Fc fragments and the formation of foam macrophages. Therefore, our findings provide a new explanation why bacterial infection deteriorates the pathogenesis of atherosclerosis.
9.Realization of dose verification tool for IMRT plan based on DPM
Jinfeng CAI ; Hui LIN ; Ruifen CAO ; Yumei DAI ; Xi PEI ; Jun ZHANG ; Gongming XU
Chinese Journal of Radiological Medicine and Protection 2014;34(5):381-385
Objective To build a Monte Carlo dose verification tool for IMRT Plan by implanting an irradiation source model into DPM code and to extend the ability of DPM to calculate any incident angles and irregular-inhomogeneous fields.Methods The virtual source and the energy spectrum unfolded from the accelerator measurement data were used,in combination with optimized intensity maps,to calculate the dose distribution of the irradiation irregular-inhomogeneous field.The irradiation source model of accelerator was substituted by a grid-based surface source.The contour and the intensity distribution of the surface source were optimized by IMRT.The dose calculation was realized by combining the position of the emitter with the fluence map from the IMRT plan.The weight of the emitter was decided by the grid intensity.The direction of the emitter was decided by the combination of the virtual source and the emitting position.The weighted fraction of the emitter was also combined with the flux grid intensity based on the particle transport model of DPM code.Results The accuracy of calculation was verified by comparing with the measured data.It was illustrated that the differences were acceptable (< 2% inside the field,2-3 mm in the penumbra).The dose calculation of irregular field by DPM simulation was also compared with that of FSPB (Finite Size Pencil Beam).The passing rate of gamma analysis was 95.1% for peripheral lung cancer.The regular field and the irregular rotational field were all within permissible range of error.The calculation time of regular fields were less than 2 h,and that of the test of peripheral lung cancer was 160 min.Conclusions The adapted DPM code with its simple irradiation source model is faster than that with classical Monte Carlo procedure.Its computational accuracy and speed satisfy the clinical requiremcnt,and it can be useful as a Monte Carlo dose verification tool for IMRT Plan.
10.Role of natural antiox-LDL subclass IgM antidodies in formation of atherclerosis
Xuyang FENG ; Bincheng REN ; Kaiyu CAO ; Ruifen XU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2018;20(3):298-301
Objective To study the role of natural antiox-LDL subclass IgM antibodies in formation of atherclerosis.Methods The macrophages from mice were divded into LDL group,Cuox-LDL group and control group.The macrophages in LDL group and Cuox-LDL group were precultured with LDL and Cuox-LDL.Adhesion experiment was performed in control group.3A6,5G8 and 2H7 were incubated.The macrophages were divided into 3A6 group,5G8 group and 2H7 group.Adhesion experiment was performed by adding the pretreated 125I Cuox-LDL.The binding parameters of macrophages and 125I Cuox-LDL in different groups were recorded.Foam cells were analyzed by cell lipid analysis with oil red O staining.Results The bound 125I Cuox-LDL level was significantly lower in Cuox-LDL group than in control group and LDL group (P<0.01).No significant difference was found in bound 125I Cuox-LDL level between 5G3 group and 2H7 group (P>0.05).The bound 125I Cuox-LDL level was significantly lower in 3A6 group than in control group,5G8 group and 2H7 group (P<0.01).Oil red O staining showed no oil drop in macrophages,but a small number of oil drops in 3A6 group and a large number of oil drops in control group.The cholesterol and TG in macrophages reduced 45% and the TG reduced 170% in 3A6 group,which were significantly lower than those in control group (P<0.01).Conclusion Antiox-LDL IgM subclass antibody 3A6 can effectively prevent the binding of unactivated macrophages to ox-LDL,reduce the occurrence and progression of atherosclerosis.