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Background Researches showed that thymic stromal lymphopoietin (TSLP) is an interleukin-17-like inflammatory factor and plays important roles in the pathogenesis and development of allergic diseases.However,the study whether TSLP plays roles in allergic conjunctivitis is rare.Objective This study was to investigate the expression change of TSLP and IL-4 in ocular surface tissue and cervical lymph node in the mice with allergic conjunctivitis induced by artemisia annua,a common plant in China,and to explore the role of TSLP and IL-4 in the pathogenesis and development of allergic conjunctivitis.Methods Twenty female BALB/c mice aged 6-8 weeks were randomized into normal control group and model group.Antigen solution was prepared using 400 μl extracting solution of artemisia annua pollen with 400 μl antigen solvent.The mouse models of allergic conjunctivitis were established by injection of 50 μl antigen solution in footpad followed by ocular topical administration of extracting solution once a day from day 10 to 12 after injection,and no any intervention was given in the normal control group.The mice were sacrificed and eyeballs were obtained 13 days after modeling,and corneal epithelium,conjunctiva and cervical lymph nodes were harvested for the detection of TSLP mRNA and IL-4 mRNA by real-time PCR.Immunochemistry was employed to assay the expression of TSLP and IL-4 proteins in corneal,conjunctival and subconjunctival tissues.Results Ocular inflammatory signs appeared 0.5 hours after ocular topical administration of extracting solution,including eyelid edema,conjunctival congestion,tears,scratching eyelids,etc.The symptoms lasted for 6-24 hours,with the model successful rate 80％.Real-time PCR indicated that the expression of IL-4 was absent in corneal epithelium in both model group and normal control group.The relative expression levels of TSLP mRNA in the corneal epithelium,conjunctiva and cervical lymph nodes in the model group were more (1.63±0.20)times,(2.71±0.48) times and (1.48 ±0.05) times than the normal control group,showing significant differences between the two groups (t =4.44,14.16,5.01,all at P＜0.05).Compared with the normal control group,the relative expression levels of IL-4 mRNA in the model group increased (2.94±0.39) times and (1.74±0.09) times,with significant differences between the two groups (t =9.92,14.54,both at P＜0.05).Immunochemistry assay showed that the expression of TSLP protein in the corneal and conjunctival epithelial cells were enhanced in the model group compared with the normal control group.In addition,an intensive expression of IL-4 protein was seen in subconjunctival tissue in the model group,while the expression of IL-4 was absent in the normal control group.Conclusions TSLP is mainly expressed in the cornea,conjunctiva and cervical lymph nodes of the mice with allergic conjunctivitis,suggesting that TSLP promotes the inflammatory process of cornea and conjunctiva;IL-4 is mainly expressed in conjunctiva,showing IL-4 participates in conjunctival inflammatory process.TSLP and IL-4 play synergistic roles in promoting the inflammatory process of ocular surface in the mice with allergic conjunctivitis,which may be new therapeutic targets.
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Background The conventional drugs for preventing and treating graft rejection have the risks of inducing adverse responses.Researches showed that resolvinE1 (RvE1) can regulate Th1 cell-mediated immunoreaction.However,whether RvE1 has an inhibit effect on high-risk corneal graft is unclear now.Objective This study was to investigate the effects of RvE1 on immune rejection in high-risk corneal grafting mouse models.Methods SPF BALB/c mice were used as recipients,C57BL/6 mice were as donors.Ninety BALB/c mice were divided into corneal allograft group,corneal allograft+RvE1 group and corneal autograft group according to random number table.High-risk corneal graft models were established by corneal suturing for 14 days and followed by penetrating keratoplasty in recipients.Allograft keratoplasty was performed on the right eyes in the mice of corneal allograft group and corneal allograft+RvE1 group,and self-corneal graft rotated 180°was transplanted on the right eyes in the mice of autograft group.Normal saline solution of 10 μl was subconjunctivally injected after surgery once per day for 7 days in the corneal allograft group and corneal autograft group,and 10 μl RyE1 (1 μg) was used in the same way in the corneal allograft+RvE1 group.The recipient eyes were examined for potential rejection signals with slit lamp microscope and calculated the mean survival time and rejection index (RI).The histopathology was examined 21 days after modeling by hemotoxylin and eosin staining.The expressions of CD4 and interferon-γ (IFN-γ)in the corneas were detected by immunohistochemistry.Th1 cell (CD3+CD8a-IFN-γ+) percentage in draining lymph nodes were measured by flow cytometry.The mRNA expression levels of interleukin-2 (IL-2),tumor mecrosis factor-α (TNF-α),IFN-γ and T-bet were detected by real-time fluorescence quantitative PCR.Results The mean survival time of grafts was (28.5± 1.7) days in the corneal allograft group,and that in the corneal allograft+RvE1 group was (14.0±1.6) days,showing a significant difference between them (t =4.14,P＜0.001),while the survival rate was 100％ at 50 days after modeling in the corneal autograft group.Corneal edema and inflammatory cell infiltration were slight in the corneal allograft+RvE1 group and corneal autograft group compared with corneal allograft group.CD4 was positively expressed in corneal tissue,and IFN-γ was expressed in corneal epithelium.The CD4+ and IFN-γ+ cell number was decreased in the corneal allograft+RvE1 group and corneal autograft group compared with corneal allograft group under the fluorescence microscope.The percentages of Th1 cells in lymph cells of corneal allograft +RvE1 group and corneal autograft group were (1.07 ±0.25) ％,(0.85 ±0.12) ％,respectively,which were significnatly lower than (1.56±0.20) ％ in the corneal allograft group (both at P＜0.05).The expressions of IL-2,TNF-α,IFN-γ and T-bet mRNA in the corneal tissue in the corneal allograft group were higher than those in the corneal allograft+RvE1 group and corneal autograft group (all at P＜0.05).Conclusions RyE1 inhibits graft rejection in high-risk allograft mouse models probably by down-regulating the Th1 cell percentage in lymph cells and the expression of inflammationrelated cytokines in corneal grafts.
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Objective: To investigate the effect of epidermal growth factoramphiregulin (AREG) on the growth of mouse colon carcinoma CT26cells and its related mechanisms.Methods: The protein expression level of AREG in different mouse cancer cells was detected by ELISA. Mouse colon carcinoma CT26 cells, melanoma B16 cellsand hepatocellular carcinoma LPC-Akt cells were transfected with the recombinant lentiviralplasmid carrying AREG gene, while the ones transfected with empty plasmid were used asthe negative controls. After AREG overexpression, the cell proliferation, colony-formingabilities and cell cycle progression in vitro were detected by MTT, colony-forming assay andFCM, respectively. After the homograft mouse model of CT26 cells was constructed, thegrowth of homograft tumor was observed, the distribution of immune cells in tumor tissueswas detected by FCM, furthermore the expression of chemokine was detected by real-timefluorescent quantitative PCR.Results: The levels of AREG expression were relatively low in mouse colon carcinoma CT26cells, melanoma B16 cells and hepatoma LPC-Akt cells. AREG overexpression did notmarkedly affect the proliferation, colony-forming abilities and cell cycle progression of thesethree types of tumor cells in vitro (all P > 0.05). However, in the homograft mouse model ofCT26 cells, AREG overexpression significantly promoted the growth of tumor cells in vivo (P <0.01), decreased the percentage of CD8+ T cells (P < 0.05), and reduced the mRNA level ofCC chemokine 5 ligand (CCL5) (P < 0.05) which was related to CD8+ T cell recruitment.Conclusion: AREG promotes the growth of mouse colon carcinoma CT26 cells in vivo . AREGmay affect the tumor microenvironment by regulating the production of chemokine which isrelated to CD8+ T cell recruitment.
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Background The pathogenesis and mechanism research of corneal neovascularization is of important significance for the prevention and management of corneal neovascularization.Some relative researches are being performed on non-corneal neovascularization-derived vascular endothelial cells, so the results are affected to a certain extent.Objective This study was to isolate and culture vascular endothelial cells from experimental corneal neovascularization tissue and detect the expression of chemokine receptors in vitro.Methods Corneal neovascularization models were established on 10 SPF male BALB/c mice with the age of 7-8 weeks by sticking the filter papers with NaOH on the central corneas, and then the immunofluorescence technique was use to assay the CD31 expression in corneal flatmount 2 weeks after modeling.Corneal pieces were made in 2 weeks after alkali burn and then were digested by collagenase type D.Vascular endothelial cells were isolated from neovascularized tissue by affinity purification using magnetic beads coated with anti-CD31.The cells were cultured on fibronectin-coated walls and then identified by immunocytochemistry.Reverse transcription-PCR was employed to detect the expressions of chemokine receptors in the cells.The use and care of the animals complied with ARVO Statement and this experimental procedure was approved by Soochow University Animal Care Committee.Results Corneal neovascularization occurred at 7 days and peaked at 2 weeks after modeling, and immunofluorescence exhibited the green network-like fluorescence for CD31 antibody in corneas.The cells grew against the wall 2 hours after culture with the polygon shape and large dimension, and the growth obviously quickened after passage.The cultured cells showed the positive response for CD31 antibody, showing the brown dye in cytoplasm,in contrast,the expression of CD31 was absent in corneal stromal cells.Chemokine receptors were positively expressed in the cells with the strongest expression levels in CCR1 ,CCR2,CCR3 and CCR4 mRNA and the weakest expression levels in CCR9,CXCR4 and CXCR5 mRNA,while CXCR3, CCR6, CCR10 and CX3CR1 mRNA were expressed with the moderate intensity.Conclusions Vascular endothelial cells can be obtained from experimental neovascularized corneas by affinity purification and express chemokine receptors,which facilitate the study of their biological properties.
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Objective To explore a simple and effective way of establishing a mice orthotopic tracheal transplantation model in studying bronchiolitis obliterans to reduce operation time. Methods Tracheas from C57BL/6 mice (n=30) were im?planted into Balb/c mice (n=30). Grafts and hosts were matched according to body weight.Orotracheal intubation and endo-stentwas used for trachea anastomose and animal model was establish. Operating time was also noted. Grafts were harvest?ed on Days 15, 30 and 60 after transplantation, using HE staining to observe pathological changes of allograft. Trachea block?ing rate was also calculated. Results Until survival of trachea transplantation, the operative time of donor′s and the recipi?ent′tracheas resection were (7.0±1.0) min and (7.0±1.0) min respectively.Orotracheal intubation the endo-stentestablish?ment cost(1 ± 0.5)min. Trachea anastomose took(20.0 ± 4.0)min and the operation time last(43.0 ± 10.0)min. Conclusion The improvement of orthotopic tracheas transplantation usingOrotracheal intubation the endo-stenfor trachea anastomose is esay-operated and did required complicate skills and instruments with high successful rate. So it is an ideal and stable model in studying bronchiolitis obliterans.
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Background Thyroid-associated ophthalmopathy (TAO) is a kind of clinically common and incurable ocular disease,and its incidence is at top place.The etiology and pathologic mechanism of TAO are still unknown because of shortness of replicative animal models and difficulty to acquire the ocular tissues in the early stage of the disease.To better understand the pathogenesis of TAO and investigate effective treatable measures, an appropriate animal model should be developed.Objective This study was to immunize female BALB/c mice with the recombinant plasmid of human thyroid-stimulating hormone receptor (TSHR) extracellular domain in cationic liposomes for the establishement of TAO models.Methods Thirty-two 6-to 8-week-old female BALB/c mice were randomly assigned to four groups according to computer random allocation.pcDNA3.1 +/hTSHR289 of 100 μg in an adjuvant cationic liposomes was injected via anterior tibialis muscle and peritoneal cavity separately in the recombinant plasmid injection group in 0, 3,6 weeks, and pcDNA3.1 or cationic liposomes was injected in the liposomes injection group or the blank plasmid group in the same way, respectively, and normal saline solution was injected in the blank control group.Body weight of the mice was measued before and 1 month,2,3 and 4 months after initial injection.The manifestations were observed after modeling.The mice were sacrificed 17 weeks after initial injection,and the histopathology examination was carried out on the thyroid gland and orbital tissue.The heart blood was collected from the mice,and serum contents of total thyroxin 4 (TT4) and thyroid-stimulating hormone (TSH)were assayed by ELISA.Results Protrusion, eyelid swell and keratitis occurred in 12 eyes of 6 mice in the recombinant plasmid injection group after immunization.A significant difference in the body weight of the mice was found among the blank control group, blank plasmid group, liposomes injection group and recombinant plasmid injection group (Fgroup =3.425, P =0.028), and the body weight was considerably reduced in the recombinant plasmid injection group in comparison with the blank control group, blank plasmid group,liposomes injection group (Ftime =0.838 ,P=0.023).The serum levels of TT4 were (7.75±1.00), (7.96±0.76), (6.76±1.10) and (4.43±2.88) μg/dlin the blank control group, liposomes injection group, blank plasmid group, and recombinant plasmid injection group, and those of TSH were (6.36±2.58),(4.83±3.96),(6.63±1.71) and (1.60 ±1.76) ng/ml, showing significant differences among the groups (F =7.150, P＜0.001;F =5.521, P＜0.01) , and the serum levels of TT4 and TSH were remarkably lower in the recombinant plasmid injection group than those of the blank control group,liposomes injection group and blank plasmid group (all at P ＜ 0.05).Histopathology revealed the lymphocyte infiltration of thyroid gland in 6 mice and proliferation of orbital adipose tissue, infiltration of lymphocytes and mastocytes,deposition of hyaluronic acid as well as swell, breakage and inflammatory cell infiltration of extraocular muscle in 15 eyes of the recombinant plasmid injection group.Conclusions A murine model of TAO can be successfully induced by immunization with recombination plasmid pcDNA3.1 +/hTSHR289 and cationic liposomes.The histopathology characteristics and ocular findings of the animal models are similar to human TAO.
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Objective To compare the pathogenicity between Ureaplasma urealyticum serotype 1 (Up1)and 8 (Uu8) in the genital tract of BALB/c mice.Methods A total of 48 BALB/c mice were randomly and equally divided into four groups:blank control group receiving no treatment,estradiol group pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with sterial liquid culture media,Up1 and Uu8 groups pretreated with intramuscular injection of estradiol followed by intravaginal inoculation with suspensions of Up1 and Uu8 respectively.Three mice were randomly selected from each group to be sacrificed after the collection of vaginal lavage fluid on day 3,7,14 and 21 after the inoculation.Vaginal and uterine tissue specimens were obtained from these sacrificed mice and underwent hematoxylin and eosin (HE) staining.Vaginal lavage fluid samples were subjected to culture of Uu and measurement of tumor necrosis factor-α (TNF-α).Results No evidences were observed for Uu growth in either the blank control group or estradiol group at any of the time points after the inoculation,with the average level of TNF-α in vaginal lavage fluid being (4.17 ± 0.85) pg/ml at these time points in both groups.Uu grew in all the vaginal lavage fluid samples from the Up1 and Uu8 groups at the four time points,with the color change unit (CCU) value decreasing with time.The level of TNF-α in vaginal lavage fluid peaked on day 14 after the inoculation in the Up 1 ((14.93 ± 1.11) pg/ml) and Uu8 ((27.04 ± 24.26) pg/ml) groups.Both Up1 and Uu8 infection caused acute and chronic inflammatory responses in the mice,which were mainly located in the uterus,and Up1 might cause intrauterine adhesion.Conclusions At the same inoculation concentration,no significant difference is found in the pathogenicity between Up1 and Uu8,both of which appear to mainly cause cervicitis.Upl might be partially responsible for intrauterine adhesion in mice.
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Objective To explore the expressions of circulating microRNAs (miRNAs) in acute liver failure mice induced by D-galactosamine (GalN)/lipopolysaccharides (LPS) and the correlation with miRNAs in the liver.Methods Forty clean grade Balb/C mice,with 32 in the model group and 8 in the control group were enrolled in the study.Liver failure was induced by intraperitoneally injection of D-GalN and LPS in mice of the model group,while mice of the control group were intraperitoneally injected with 1 mL 0.9 ％ sodium chloride solution.Serum and liver samples were collected at 0,3,5,7 hours following administration,and eight mice should be supplied to each sample,and changes of alanine aminotransferase (ALT),aspartate aminotransferase (AST) and histopathology of the liver were observed.miRNA from both the serum and the liver was extracted,miRNA expression profile in the liver at 0,5,7 hours by locked nucleic acid (LNA)-miRNA microarray was analyzed and miRNA by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) was detected.Means of the two groups were compared using one-way ANOVA and correlation analyses were performed using Pearson and Spearman correlation.Results Expression of miRNAs in the liver tissue changed significantly over time with the occurrence of acute liver failure in the mice.Twenty-one miRNAs were up-regulated and 27 were down-regulated,among which miRNA-122 and miRNA-1187 were down-regulated while miRNA-146a and miRNA-155 were up-regulated.It was confirmed by the PCR assay that the expression of miRNA-122 and miRNA-1187 in the liver gradually decreased,while those in the serum were up-regulated over time.However,the expressions of inflammation associated miRNA-155 and miRNA-146a were up-regulated both in the serum and the liver after administration.The expressions of miRNA-122 and miRNA-1187 were negatively correlated between serum and liver (r=-0.477,P=0.0089,r=-0.420,P=0.231),while the expressions of miRNA-155 in serum and liver were positively correlated (r=0.678,P=0.0001).Moreover,the expressions of miRNA-122 (r=0.571,0.554) and miRNA-1187 (r=0.471,0.542) were also positively correlated with serum levels of ALT and AST (all P＜0.05).Liver and serum levels of miRNA-122 and miRNA-1187 changed significantly at 5 hours after administration,which preceded the changes of ALT/AST.Conclusions The expressions of miRNA-122 and miRNA-1187 in serum are well inversely correlated with the corresponding expressions in liver tissues during acute liver failure in mice.The changes of miRNA-122 and miRNA-1187 in the serum precede those of ALT/AST.These data suggest that serum miRNA-122 and miRNA-1187 might be the candidate serum biomarkers for early prediction of liver injury.
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Objective To explore the effects of low molecular weight heparin (LMWH) on the virological and pathological changes of newborn mouse liver congenitally infected with human cytomegalovirus (HCMV).Methods Sixty healthy pure line clean level BALB/c mice which were about 10 weeks old (half were female) were divided into five groups (six pairs in each group).The mice in LMWH intervention group and positive control group were intraperitoneally inoculated with 6.0 lg tissue culture infective dose50 (TCID50) of HCMV AD169; those in blank control group were intraperitoneally injected with 0.5 mL dulbecco's modified Eagles medium (DMEM) ; then all the mice were paired to mate.The pregnant mice in LMWH intervention group Ⅰ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days; those in group Ⅱ were subcutaneously injected with LMWH 1000 U/kg daily for 5 days and their newborn mice were subcutaneously injected with LMWH 1000 U/kg daily for 5 days; those in group Ⅲ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days in their newborn mice.All these newborn mice were sacrificed at day 10 of birth.The liver was removed for virus isolation,dry-wet weight determination,pathology examination and quantitative fluorescence-polymerase chain reaction (PCR) detection.The comparison among groups was done by analysis of variance.Results HCMV was isolated from the supernatant of liver tissue homogenate in 10-day positive control newborn mice,while HCMV was isolated in 24-day newborn mice of the other three groups of LMWH intervention.Pathology confirmed that positive control liver tissue had inflammatory changed,liver cell inflammatory swelling degeneration,vacuoles degeneration,specific HCMV inclusion body in nuclear,and portion of liver cell necrosis,while liver pathological results of LMWH intervention group showed mild liver cell inflammatory changes and slight cell inflammatory swelling degeneration,which were similar to the blank control group.The moisture of liver tissue contents in LMWH intervention group decreased more obviously than positive control group.The HCMV DNA loads in 50 mg liver tissues of LMWH intervention groups Ⅰ,Ⅱand Ⅲ were (3.26±0.43),(3.26±0.41) and (3.32±0.51) lg copy,respectively,which were significantly lower than that of positive control group [(7.38 ± 0.53) lg copy; F =314.620,P0.01],while there were no significant differences among LMWH intervention groups (P＞0.05).Conclusion LMWH intrauterine and postnatal interventions can significantly reduce HCMV DNA replication in hepatocytes,and relieve inflammatory changes in liver tissue.
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ObjectiveTo obtain stable animal models and observe Helicobacter hepaticas (Hh)colonization and pathological claracteristics,through infecting different mice strains with Hh.Methods SPF-class male BABL/c Cr,SCID/Cr and C57BL/6 Cr mice were inoculated 0.2 ml Hh standard strain ATCC51450 bacterial suspension (1 × 108CUF/ml),inoculated for 3 times with 48 hours intervals,the control group was fed with the same volume of PBS.Mice were executed at 4 weeks,8weeks and 16 weeks since last Hh inoculation,and mice esophagus,stomach,jejunum,ileum,cecum,colon,liver and pancreas tissue were taken for histopathology examination,Micro-aerobic bacteria isolation,culture and identification and Hh specific 16S rRNA gene amplification.Results The colonization rates of Hh in cecum after inoculated in BALB/c Cr mice and SCID/Cr mice at 4 weeks,8 weeks and 16 weeks were all 8/8,colonization rates in colon at 4 weeks,8 weeks and 16 weeks were 4/8,5/8,5/8 and 3/8,6/8,5/8 respectively,colonization rates in ileum and jejunum at 16 weeks were 1/8,colonization rates in liver at 8 weeks and 16 weeks were 2/8,3/8 and 2/8,2/8respectively.The colonization rates of Hh in cecum after inoculated in C57BL/6 Cr mice at 4 weeks,8weeks and 16 weeks were 1/8,2/8 and 2/8 respectively,colonization rates in colon at 8 weeks and 16weeks were 1/8,2/8 respectively.Compared with C57BL/6 Cr mice,the inflammatory changes in liver,cecum and colon were more significant in Hh infected BALB/c Cr and SCID/Cr mice (P＜0.01),and histological scores gradually increased as infection time extended (P＜0.05,P＜ 0.01 ).The histological scores were significantly higher in those with colon and liver Hh bacterial colonization than those without Hh bacterial colonization (P＜0.05).The histopathological score of cecal tissue was positively correlated with the density of Hh colonization.ConclusionDifferent mice strains are with different susceptibility to Hh,and better Hh infection model can be obtained in Hh inoculated BALB/c Cr and SCID/Cr mice.
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Objective:To investigate the effects of endostatin on Ang-2/Tie-2 system in ectopic lesion.Methods:Thirty mice of endometriosis model (EM) were established by peritoneum planted.After 1 week of transplantation,model mice were randomly divided into three groups including rAAV2-hEndostatin-EGFP group (n=20),rAAV2-EGFP group (n=20)and PBS control group (n=20).The histology changes were examined in the three groups.The expressions of Ang-2 and Tie-2 Were examined by immunohistochemistry.Results:The EM model mice were established suecessfully.The gland proliferation was obvious in the two control groups.The mesenchymal blood vessel was rich.But there were gland atrophy and blood vessel reducing in endostatin group.There were expressions of Ang-2 and Tie-2 in the cytoplasm of endothelium,epithelial glands and stroma.The positive rate of Ang-2 expression was 40%in endostatin group,much lower than that in other two groups(75%and 80%,P＜0.05).The positive rate of Tie-2 expression was 45%in endostatin group,which was also lower than that in other two groups(80%and 85%,P＜O.05).Conclusion:Endostatin can effectively interfere angiogenesis process of Ang-2/Tie-2,which lays the foundation for clinical therapy by anti-angiogenesis.
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Objective: To observe the biological difference of multi-drug resistant nasopharyngeal carcinoma (NPC) cell line CNE2/DDP and its parental cell line CNE2, as well as analyze the characteristics of multi-drug resistant cells. Methods: The difference in morphological features, growth characteristics and colony formation rate in vitro were examined and compared under light microscope. Flow cytometry was used to evaluate the cell cycle distribution and the expression of P-170 and ABCG2 proteins. Drug sensitivity was measured by MTT assay. Tumorigenicity was evaluated after subcutaneous injection of 1 × 106 CNE2/DDP or CNE2 cells into BALB/c nude mice. Results: Under light microscope CNE2/DDP cells were smaller and showed a round shape and the CNE2 cells appeared polygonal cell bodies. CNE2/DDP cells had relatively longer colony doubling time than CNE2 cells (29.46 h vs 21.03 h) indicating that the growth of CNE2/DDP cells was slower. There was significant difference in the cell cycle distribution between CNE2/DDP and CNE2 cells [ (65.12 ± 1.67) % vs (44.9 ± 2.2) % in G0/G1 phase, (23.63 ± 0.42)% vs (39.67 ± 1.27) % in S phase, and (10.93 ± 0.25) % vs (13.23 ± 0.31)% in G2/M phase, P < 0.01]. Expression of P-170 and ABCG2 proteins in CNE2/DDP cells were significantly higher than that in CNE2 cells [(75.93 ± 0.86)% vs (2.83 ± 0.40)%, (43.37 ± 0.42) % vs (4.07 ± 0.59)% P < 0.01]. There was no significant difference in the resistance indexes of CNE2/DDP cells to DDP,5-FU and VCR before and after transplantation. The tumorigenicity time was (17.17 ± 1.17) d and (10.00 ± 2.68) d respectively for CNE2/DDP and CNE2 cells after being transplanted into BALB/c nude mice. Conclusion: CNE2/DDP has typical multi-drug resistance features and could be used for the research of drug-resistant NPC.
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Objective To study the efficacy and mechanism of compound Tripterygium hypoglaucum Hutch (THH) on photoallergic contact dermatitis in mice. Methods The photoallergic animal model of BALB/c mice was established by using photosensitizer chlorpromazine and UVA irradiation. The therepeutic efficacy was determined by measuring the thickness and the weight of the swelling ear and the number of infiltrated mononuclear cells in the ear tissue. Immunohistochemical technique was used to detect the ICAM-1 expression on keratinocytes, fibroblasts and vascular endothelial cells. The serum level of INF-? was measured by ELISA. The tested animals were divided into 3 groups: compound THH, THH alone and normal saline. Results The difference of the thickness of left ear before and after challenge, the differences of the thickness and the weight of ear tissue, the difference of the number of infiltrated mononuclear cells of left and right ear after challenge were significantly less in the compound THH group than those in the THH alone group (P
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The present experiment was to study the hematopoietic reconstruction effects of peripheral blood stem cell (PBSC) mobilized by anti-CD49d monoclonal antibody (McAb) and recombinant human granulocyte colony-stimulating factor(rhG-CSF) in mice. PBSCs from NS-treated mice (control group), rhG-CSF-mobilized mice (experimental group1), and anti-CD49d McAb-mobilized mice (experimental group 2), The changes in respectively, were transplanted to BALB/c mice pre-conditioned with high-dose chemotherapy and total body irradiation white blood cell (WBC) count, four-weeks survival rate, bone marrow nuclear cells (BMNC), granulocyte-macrophage colony-forming units (CFU-GM), and colony-forming unit-spleen (CFU-S) were observed. The results showed that survival rate, WBC, BMNC, CFU-GM, and CFU-S counts were significantly higher in experimental groups 1 and 2 than those of the control group(P
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Uno de los pilares fundamentales en el control, la eliminación y posible erradicación de la hepatitis viral tipo B-D es la utilización de la vacuna de producción cubana contra la hepatitis B. Se desarrollaron modelos experimentales y aplicados al humano que permitieron el estudio de inmunogenicidad mediante distintas vías, dosis y esquemas. Se trabajó experimentalmente se trabajó con ratones BALB/c, con la aplicación de 1 mg/dosis por vía intramuscular (IM) e intraperitoneal (IP) en 3 esquemas diferentes 0-1-2, 0-1-4 y 0-4-8 semanas. La inmunogenicidad se estudió mediante la cuantificación del anticuerpo contra el antígeno de superficie (AgsHB), con la aplicación de un método inmunoenzimático. En todos los animales hubo seroconversión con los esquemas 0-1-4 y 0-4-8 semanas. En el esquema 0-1-2 la vía IP resultó más inmunogénica que la vía IM, produjeron 60 y 35 % de seroconversión, respectivamente. La vía IP tuvo un coeficiente de variación menor cuando se compararon los esquemas 0-1-4 y 0-4-8 semanas, de todas formas resultó alto para ser animales BALB/c. En humanos se inmunizaron estudiantes de medicina y enfermería con el empleo de 20 mg/dosis por vía IM y 2 mg/dosis vía intradérmica (ID), con los esquemas 0-1-4 y 0-4-8 semanas; se demostró que la seroprotección e hiperrespuesta era mayor en el esquema 0-4-8 independientemente de la vía utilizada y que el esquema 0-1-4 no produjo la inmunogenicidad requerida por lo que debe ser utilizado de manera excepcional. Se obtuvieron estándares de anticuerpos nacionales para la aplicación en métodos de cuantificación de anti-AgsHB, tanto en estudios experimentales como en humanos.
One of the fundamental pillars in the control, elimination and possible eradication of B-D viral hepatitis is the use of the Cuban-made hepatitis B vaccine. Experimental models as well as models for humans were developed to study immunogenicity using various routes of administration, doses and schedules. Balb/c mice were used on experimental basis to which 1 mg/dose were administered by intramuscular(IM) and intraperitoneal (IP) within three different vaccination schedules, i,e, at 0, 1, and 2 weeks; 0, 1 and 4 weeks, and 0, 4 and 8 weeks. Immunogenicity was stufied through quantification of antibodies to surface antigens by using an immunoenzymatic method. Seroconversion was present in all animals at 0, 1 and 4 and 0, 1, and 8 weeks schedule. However, in 0, 1 and 2 schedule, IP route of administration was more immunogenic than IM, with 60% and 35% seroconversion respectively. IP route had a lower variation coefficient than 0, 1 and 4 week and 0, 4 and 8 schedules were compared, but anyway, it was high even for Balb/c animals. Medical and nursing students were immunized with 20 mg/dose IM and 2mg/dose intradermally within 0, 1 and 4 week and 0,4 and 8 w schedules. It was showed that seroprotection and hyperesponse were greater in 0, 4 and 8 week vaccination schemes regardless of the route of administration and that 0, 1 and 4 w schedule does not bring required immunogenicity, therefore, it should be used exceptionally. National antibody standards were obtained to be applied in anti-HbsAg quantification methods both in experimental and human-based studies.
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Se explora el efecto de la administración de 7 mg/kg de peso corporal diarios de teofilina por vía intraperitoneal entre los días 8 y 14 de la preñez en 28 ratones BALB/C sobre el contenido de DNA y proteínas de los fetos al nacer y los estimadores del tamaño y número de células. Los resultados fueron comparados con los de un grupo control de 36 animales. Los resultados indicaron que la teofilina no afectó significativamente el contenido de DNA y proteínas, ni el tamaño y número de células de los fetos.
The effects of the administration of 7 mg/kg of body weight of daily intraperitoneal theophylline, is explored between days 8 and 14 of pregnancy in 28 BALB/C mice, on the DNA and protein contents of the fetuses at birth, and the estimators of size and cell amount. The results were compared with those from a 36 animals control group. Those results indicated that theophylline didn't significantly affect either the DNA and protein contents, or the size and cell amount of the fetuses.
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The expression of estrogen receptor (ER) in thymus of female BALB/c mice at different ages was investigated. The results showed that the expression of ER in thymus was increased from 1 to 8 weeks after birth. ER positive cells were thymic epithelial cells and thymocytes. ER existed not only in the cytoplasm, but also in the nucleus.