2.Effect of allergen on function of peripheral blood CD4+T cells in patients with asthma
yin-shi, GUO ; yi-ping, XU ; gui-ying, SHI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(06):-
Objective To investigate the relationship among the proliferation of CD4+T cells, the intracellular levels of interleukin-10 (IL-10) and transforming growth factor-?1(TGF-?1), and allergic bronchial asthma. Methods Dermatophagoides farinae antigen were prepared as allergen. Twenty-five patients with asthma and 15 healthy individuals were enrolled and divided into blank control group, allergen group and self control group, respectively, after venous blood sample collection. The proliferation of CD4+T cells and the distributions of CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 were measuredby flow cytometry (FCM). Results The distributions of CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 could hardly be detected in the peripheral blood samples of the blank controls of the patients with asthma and healthy ones. In the allergen group of the healthy individuals, the peripheral blood CD4+T cells were significantly proliferated, and the proportions of CD4+T cells andCD4+/IL-10+ cells were much higher than the self control group, while there was no significantly increase in the proportions of CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 subgroups. In the allergen group of those with asthma, the proportions of peripheral blood CD4+, CD4+/IL-10+, CD4+/TGF-?+1 and CD4+/IL-10+/TGF-?+1 cells were not found significantly increased compared with those self controls. After being activated by allergen, the proportion of peripheral blood CD4+/IL-10+ cells was significantly lower in the patients with asthma than the healthy individuals(P
3.Effects of Albizia Julibrissin Flower Total Flavonoids on Learning and Memory Abilities and Plasma Monoamine Neurotransmitters of Depression Model Rats
Chaofeng GUO ; Shenggao YIN ; Meng XIA ; Xueli SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(6):1318-1322
This study was aimed to observe effects of Albizia Julibrissin Flower Total Flavonoids (AJFTF) on the learning and memory abilities and plasma monoamine neurotransmitters 5-HT and NE content of depression model rats . A total of 90 SD rats were randomly divided into the normal group , model group , western medicine group ( Venlafaxine Hydrochloride 0 . 0125 g?kg-1 ) and high , middle and low dosage groups of AJFTF ( equivalent to the crude drug of 10 , 5 , 2 . 5 g?kg-1 ) . Depression model rats were induced by solitary cultiva-tion and chronic unpredictable stress . The learning and memory abilities of model rats were evaluated with the Morris water maze . And the plasma contents of 5-HT and NE were determined by enzyme-linked immunosor-bent assay ( ELISA ) . The results showed that AJFTF ( administered with high , middle and low dosage for 21 days ) significantly shortened the incubation period of the Morris water maze positioning and navigation ( P <0 . 05 or P < 0 . 01 ) , and increased the times of crossing the platform in space exploration ( P < 0 . 05 or P <0 . 01 ) . There was a certain dose-effect relationship . The AJFTF ( high , middle and low dosage ) increased the plasma contents of 5-HT and NE ( P < 0 . 05 ) . It was concluded that AJFTF can improve learning and memory abilities of depression model rats which indicates that AJFTF have potential antidepressant effects . Its action mechanism may be related to increase plasma monoamine neurotransmitter content of 5-HT and NE .
4.Effects of acupuncture and moxibustion on chromatin remodeling-related enzymes in the colon tissue of rats with Crohn disease
Yanting LI ; Yajing GUO ; Jimeng ZHAO ; Yi SUN ; Sen GUO ; Yin SHI ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(5):353-362
Objective:To investigate the regulatory effects of acupuncture and moxibustion on chromatin remodeling complex core catalytic subunit of Brahma-related gene 1(Brg1),histone deacetylase(HDAC)3,HDAC9,and males absent on the first(MOF)in the colon tissue of rats with Crohn disease(CD). Methods:Using the random number table method,60 male Sprague-Dawley rats were divided into 5 groups,including a normal group,a model group,an acupuncture group,a medicinal cake-insulated moxibustion group,and an acupuncture-moxibustion group,with 12 rats in each group.CD rat models were prepared using 2,4,6-trinitrobenzene sulfonic acid(TNBS)in all groups except the normal group.The normal and model groups received no interventions.In the acupuncture group,rats were intervened with acupuncture at bilateral Zusanli(ST36)and Shangjuxu(ST37),20 min/session,once a day.The medicinal cake-insulated moxibustion group received medicinal cake-insulated moxibustion at Qihai(CV6)and bilateral Tianshu(ST25)with 2 cones per point per session,once a day.The acupuncture-moxibustion group received both acupuncture and moxibustion interventions simultaneously.Each intervention was performed for 10 consecutive days.Observations included general condition,disease activity,macroscopic damage,and pathological changes in the rat's colon tissue.Real-time fluorescence quantitative polymerase chain reaction was used to measure the mRNA expression of chromatin remodeling-related enzymes Brg1,HDAC3,and HDAC9,while Western blotting detected the protein expression of Brg1,HDAC3,HDAC9,and MOF in rat's colon tissue. Results:The model group showed significantly increased diarrhea score,occult blood score,macroscopic damage score of colon tissue,and colon macroscopic damage index(CMDI)score,as well as elevated mRNA expression levels of HDAC3 and HDAC9,protein expression levels of HDAC3,HDAC9,and MOF,and decreased mRNA and protein expression levels of Brg1 compared to the normal group(P<0.01).In contrast,compared to the model group,the diarrhea score,occult blood score,macroscopic damage score,CMDI score,mRNA expression levels of HDAC3 and HDAC9,and protein expression levels of HDAC3,HDAC9,and MOF were significantly reduced(P<0.05 or P<0.01),while the mRNA and protein expression levels of Brg1 were significantly increased(P<0.01)in the acupuncture group,the medicinal cake-insulated moxibustion group,and the acupuncture-moxibustion group. Conclusion:Both medicinal cake-insulated moxibustion and acupuncture,either used alone or in combination,can regulate the abnormal expression of chromatin remodeling-related enzymes Brg1,HDAC3,HDAC9,and MOF in the colon tissue,thus reducing colon inflammation in CD rats.
5.Human endostatin antiangiogenic gene therapy mediated by recombinant adeno-associated virus vector in nude mouse with endometriosis
Junjie SUN ; Lirong YIN ; Ruoran MI ; Hongda MA ; Sujie GUO ; Yang SHI ; Yanjun GU
Chinese Journal of Obstetrics and Gynecology 2010;45(1):45-50
Objective To study the therapeutic effect of recombinant adeno-associated virus carrying human endostatin gene therapy on endometriosis in mice model. Methods Recombinant adeno-associated virus vector carrying human endostatin gene and enhanced green fluorescent proteins gene (rAAV2-endostatin-EGFP) was constructed. Endometrium was from 12 patients with leiomyoma undergoing hysterectomy in Second Hospital, Tianjin Medical University between November and December 2008. Endometriosis models of nude mice were established by transplanting human endometrial fragments intooperitoneal surface. After 1 week, those 60 mice were divided into 3 groups: treatment group including 20 mice injected with rAAV2-endostatin-EGFP to ectopic lesion, control group including 20 mice injected with rAAV2-EGFP to ectopic lesion and blank control group including 20 mice injected with phosphate buffered saline (PBS) to the ectopic lesion. At 1, 2 and 3 weeks after treatment, those mice underwent laparotemy to observe the location and size of ectopic lesion in abdominal cavity. The expression of endostain protein, number of gland, microvessel density (MVD) and vascular endothelial growth factor (VEGF) were measured in ectopic lesions. The serum level of estradiol and progesterone were detected in nude mice among every groups. Results (1) All endometriosis of nude mice models were established successfully through peritoneum transplanting. After 1 week's treatment, flat lesion nodes, decreased gland number and narrow and atrophy glandular cavity were observed by light microscope. (2) The endostatin gene was transferred into nude mice successfully and expressed effectively. It was observed that endostatin protein expression was shown with enhanced green fluorescent proteins in ectopic lesion. (3) Glands number of ectopic lesion in rAAV2-endostatin-EGFP group(7.8±1.9,7.0±1.5 and 5.5±1.7) were significantly less than 10.1± 1.7, 10.2±2.0 and 9.8±2.4 in rAAV2-EGFP control group and 10.2±2.2,10.0±2.0 and 9.7±2.2 in PBS control group at 1,2 and 3 weeks after treatment(all P<0.05). Glands number of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (4) MVD of ectopic lesion in rAAV2-endostatin-EGFP group (12.2±1.5,11.4±2.1 and 9.0±1.4) was significantly less than those at rAAV2-EGFP control group (16.5±1.7,16.5±1.9 and 16.9±1.9) and PBS control group (16.2±1.6,16.0±1.6 and 16.3±1.7) at 1,2 and 3 weeks after treatment (all P<0.05) . MVD of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment(P<0.05). (5) The rate and density of VEGF expression at ectopic lesion in rAAV2-endostatin-EGFP group (35%, 30%, 25% and 1.60±0. 43,1.33± 0. 30,1.03±0.36) were significantly less than those at rAAV2-EGFP control group (80% ,75% ,85% and 2.43±0.53,2.43±0.29,2.66±0.45) and PBS control group (85% ,90% ,90% and 2.36±0.53,2.64± 0.57,2.53±0.52) at one 1, 2 and 3 ,weeks after treatment (all P<0.05). The expression of VEGF at ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (6) The level of estradial and progesterone in serum of nude mice of rAAV2-endostatin-EGFP group [ E_(2)> : (48±7 ) pmol/L, P: (61±8 ) nmol/L ] did not reach statistical difference when compared with those at rAAV2-EGFP control group [ E_(2): (50±9) pmol/L, P: (60±10) nmol/L] and PBS control group [E_(2):(48±7)pmol/L,P: (58±10)nmol/L,P>0.05]. Conclusions The recombinant adeno-asseciated virus carrying human endostatin gene therapy could inhibit angiogenesis at endometriotic lesions and not influence steroid level. The antiangiogenic gene therapy might become a novel option for endometriosis.
6.Evaluation of the vaccinations with three strains of gene-deleted mutants from pseudorabies virus, PRV TK-, PRV gE-/gI and PRV TK-/gE-/gI- after exposure to the wild Fa strain
Qigui YAN ; Aiguo YANG ; Wanzhu GUO ; Zhiwen XU ; Qian SHI ; Huaping YIN
Chinese Journal of Zoonoses 2008;(9):827-835
To evaluate the value of the vaccinations with 3 strains of gene-deleted mutants from pseudorabies virus(PRV), PRV TK-, PRV gE-/gI- and PRV TK-/gE-/gI- after to exposure to the wild Fa strain, these mutant strains from the PVR reference isolate Fa were used to vaccinate 4 weeks old PRV-free pigs with a dosage of 105 PFU each ,and followed by nasally challenged by the parental Fa strain with a dosage of 107 PFU at 14 days post vaccination. The pathological changes, virus discharge and distribution were evaluated after vaccination and challenge. It was found that the histopathological observations in the 10 collected samples including cerebrum, cerebellum, heart, liver, lungs, spleen, kidneys, tonsils, lymph nodes and trigeminal ganglion from these 3 mutant strains showed that the rates of occurrence of pathological changes in various organs were 4/10, 3/10 and 4/10 respectively, whereas that of the positive controls were 9/10. The damage in lungs was more serious in pigs vaccinated with PRV TK-mutant and positive control in comparison with other groups of pigs inoculated, and the damages in cerebrum, cerebellum and trigeminal ganglion in positive controls were more serious than those of pigs vaccinated with the 3 gene-deleted mutants. However, the tonsils, the main organ for latent infection were damaged mildly in the pigs inoculated with these 3 gene-deleted mutants in comparison with that of the positive controls. As demonstrated by Southern blot analysis, all the vaccinated pigs could discharge viruses by secretion through nasal cavity, but the soldier pigs were not infected successively by the gene-deleted mutants and the gene-deleted mutants were also unable to establish infection in cerebrum and cerebellum. Nevertheless, they could not effectively block discharge of PRV Fa after exposure to Fa virus, but could block effectively the virulent Fa virus invading into cerebrum and cerebellum. From these observation, it is evident that the deleted mutants of the TK, gE/gI , TK/gE/gI genes can block the invasion of virulent Fa virus into cerebrum and cerebellum and lessen the damages on multiple organs or tissues ,indicating that the deleted mutant of TK/gE/gI gene may be the most promising candidate of vaccine strain for development of the commercial vaccine.
7.Effect of different meal replacements in patients with metabolic syndrome
Yin HAN ; Yan LIU ; Jieli SHI ; Ruixia LI ; Sufen LI ; Nana JIN ; Honglei GUO
Chinese Journal of Endocrinology and Metabolism 2015;31(3):215-218
Objective To investigate the impact of preload fasting and meal replacement in patients with metabolic syndrome.Methods A total of 92 subjects with metabolic syndrome were enrolled in the study.They were assigned into the preload fasting group (PFG),the meal replacement group (MRG),and the control group (CG) for 12-weeks intervention.Special dietary with 100 kcal was provided 30 min before each meal in the PFG,and while in the MRG the same dietary was taken just before each meal and the amount of meal was reduced appropriately.The subjects in CG took meals as usual.Body mass index,waist circumference,and insulin resistance were assessed.Satiety situation was investigated by the scale.Results After 12 weeks,improvement were found in fasting insulin(-3.29 mU/L) and waist circumference (-4.04 cm) in the PFG and significant difference was shown compared to the CG (P<0.05).Satiety index in the PFG was the most significant among the three group.Conclusion Preload fasting is helpful in improving insulin resistance,reducing waist circumference,and enhancing satiety.
8.Transcription factor SP1 decoy ODNs targeting α1, 3-GT renders porcine endothelial cells resistant to complement-mediated cytotoxicity
Yabing HUANG ; Lu WANG ; Zhuzeng YIN ; Rong LI ; Hui GUO ; Shi CHEN
Chinese Journal of Organ Transplantation 2008;29(10):594-597
Objective To investigate whether porcine endothelial cells transfected with SP1 de-coy ODNs could resist complement-mediated cytotoxicity during the model of SV-40-PED with human serum in vitro. Methods Immortalized porcine aortic endothelial cells of the line PED were divided in-to three groups. The transfected group was SV-40-PED with SP1 decoy ODNs. The mismatched group was SV-40-PED with scrambled SP1 decoy ODNs. The negative group was cells with oligo-fectamine only. The expression of α1,3-GT mRNA and αGal was detected after 26 h by using fluores-cence microscope, Western blot, RT-PCR and lactate dehydrogenase (LDH) activity assay. Results Fluorescence microscopy observed the decreased fluorescence of αGal after SP1 decoy ODNs transfec-tion. Dotted fluorescent decrease could be observed on some membrane while the mismatched group and negative group with bright green fluorescence. Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was decreased to 52.6% of the negative group (P<0.05). The expression of α1,3-GT mRNA in the PED cells transfected with decoy ODNs was 0.42±0.20 (isoform 1) and 0.27±0.12 (isoform 2) respectively, significantly lower than in the negative group (isoform 1, 0.72±0.17; isoform 2, 0.50±0.19; both P<0.05). The expression of α1, 3-G Tin the mismatch group was not different from that in the negative group (P>0.05). 20% normal hu-man serum (NHS) and 40 % NHS had cytotoxic effect on both mismatch and negative groups, but de-coy ODNs could confer SV-40-PED protection from the cytolysis effect (P<0.05), which made a re-markable reduction of complement-mediated cytotoxicity towards SV-40--PED. Conclusions SP1 decoy ODNs could confer porcine endothelial cells protection from complement-mediated cytotoxicity effect in vitro.
9.Moxibustion on Telomerase Activity in Aging Rat
Huangan WU ; Lanqin GUO ; Hanping CHEN ; Guojie SUN ; Wei ZHANG ; Yin SHI ; Qiujuan ZHANG ; Huirong LIU
Journal of Acupuncture and Tuina Science 2007;5(2):74-78
Objective: To investigate the effect of moxibustion on telomerase activity and genes expression in tissues of senescent rats. Methods: Subacute aging rats model were established by injection with D-gal solution. Points Shenshu (BL 23) were treated with moxibustion in treatment group, contrasting with a model group and a normal group. Enzyme-Linked Immunosorbent Assays(ELISA) was used for the level oftelomerase activity in liver tissues, and In Situ Hybridization(ISH) was used for the condition of expression of telomerase genes in liver tissues. Results: The level of telomerase activity in the aging model group was obviously lower than that in normal control group (P<0.01), the level in moxibustion group was obviously higher than that in model group(P<0.05). In comparison with normal rats, the positive-expressed areas and photodensity of telomerase genes in aging model group were all significantly lower (P<0.01,P<0.01), and the positive-expressed areas in moxibustion group was significantly higher than that in model group (P<0.01). Conclusions: Moxibustion could regulate telomerase activity of senile rats, hence delaying aging.