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Abstract Introduction: This study's objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats. Methods: Sprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry. Results: The expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1β, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group. Conclusion: Downregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.
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OBJECTIVE@#To compare the clinical effect between wheat-grain moxibustion at Yinbai (SP 1) and oral administration of dydrogesterone tablet for menstrual period prolongation after down-regulation treatment of in vitro fertilization embryo transfer (IVF-ET).@*METHODS@#A total of 54 patients with prolonged menstrual period after down-regulation treatment of IVF-ET were randomly divided into an observation group and a control group, 27 cases in each one. In the observation group, when the menstrual period delayed more than 7 days, the wheat-grain moxibustion at Yinbai (SP 1) was performed, once a day, with an interval of 1 day between two 3-day treatments; when the menstrual blood was cleaned, the ovulation was continued and the eggs were taken. In the control group, when the menstrual period delayed more than 7 days, the oral administration of dydrogesterone tablet was provided, 10 mg each time, twice a day; when the menstrual blood was cleaned, the ovulation was continued and the eggs were taken. The number of days for menstrual blood to be cleaned, the area change of uterine cavity hemorrhage, the morphology of endometrium, the blood supply of endometrium, the number of oocytes obtained, the grade of frozen embryo and the clinical effect were observed between the two groups after treatment.@*RESULTS@#Compared with the control group, the number of days for menstrual blood to be cleaned was shorter in the observation group after treatment (0.05). The cured rate in the observation group was 100.0% (27/27), higher than 33.3% (9/27) in the control group (<0.05).@*CONCLUSION@#The wheat-grain moxibustion at Yinbai (SP 1) could more effectively treat prolonged menstrual period after IVF-ET down-regulation treatment, which is beneficial to the preparation of the endometrium, and has no effect on the oocyte collection and embryo culture.
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@#【Objective】To investigate the effect of adding different preparations containing luteinizing hormone activity in patients with normal ovarian reserve but suboptimal response during GnRH agonist protocol when undergoing IVF/ICSI- ET.【Methods】872 infertile patients with normal ovarian reserve but suboptimal response to FSH during GnRH agonist protocol were enrolled. According to the supplementation of different preparations containing LH activity ，patients were divided into three groups，including low- dose hCG group（n=28），rLH group（n=319）and HMG group（n=525）. The clinical parameters and pregnancy outcomes were retrospectively compared among three groups.【Results】The proportion of ultra-long GnRH-a protocol in hCG group was higher than that in HMG group（14.3% vs. 1.1%，P＜0.001）. The total duration and dosage gonadotrophin in hCG group were more than that in HMG group ［15.0（13.0~16.8）vs. 13.0（12.0~15.0）days ，P = 0.027 ；2 925（2 531~3 900）vs. 2 550（2 100~3 225）U ，P = 0.046］. The total duration and dosage gonadotrophin in rLH group were less than that in HMG group［13.0（12.0~14.0）vs. 13.0（12.0~15.0）days，P = 0.009；2 400（1 950~3 075）vs. 2 550（2 100~3 225）U ，P = 0.009］. There were 53.6%（15/28）patients who still showed suboptimal response after the administration of HMG or rLH in hCG group. The clinical pregnancy rate（69.2%，58.6% vs.63.8%；P＞0.05）and live birth rate（65.4%，49.6% vs. 53.1%；P＞0.05）were similar among these groups.【Conclusions】 For patients with normal ovarian reserve but suboptimal response during GnRH agonist protocol ，the supplementation of different preparations containing LH activity showed comparable effect on pregnancy outcomes. The addition of low- dose hCG was effective even when patients still showed suboptimal response after the administration of HMG or rLH.
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Wilms tumor 1 (WT1) has long been overexpressed in acute myeloid leukemia and has a prognostic value in its diagnosis.Lately, the formation of G-quadruplexes in oncogenic promoters like WT1 has been widely investigated since stabilizationof these structures leads to transcriptional inhibition of the oncogene. Daunorubicin and mitoxantrone considered as crucialcomponents of almost all standard acute myeloid leukemia induction regimens. Herein we have proposed a probablemolecular mechanism of action through which the drugs may stabilize WT1 promoter G-quadruplexes. Differential pulsevoltammetry, circular dichroism, and polyacrylamide gel electrophoresis, electrophoretic mobility shifts assay, polymerasechain reaction (PCR) stop assays, and quantitative RT-PCR were performed in order to better understanding the nature ofinteractions between the drugs and G-quadruplexes. Data revealed that both drugs had potential to stabilize G-quadruplexesand down-regulate WT1 transcription but daunorubicin exposed more silencing impact. The results illustrated the therapeutic association of these two commercial FDA-approved drugs in WT1 transcriptional down-regulation. Since WT1 hasknown as a transcriptional regulator of at least 137 target genes, so the new data are significant for the development of newapproaches to regulating WT1 and other target genes by employing special drugs in cancer treatment.
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BACKGROUND/AIMS: Interstitial cells of Cajal (ICC) and their special calcium-activated chloride channel, anoctamin-1 (ANO1) play pivotal roles in regulating colonic transit. This study is designed to investigate the role of ICC and the ANO1 channel in colonic transit disorder in dextran sodium sulfate (DSS)-treated colitis mice. METHODS: Colonic transit experiment, colonic migrating motor complexes (CMMCs), smooth muscle spontaneous contractile experiments, intracellular electrical recordings, western blotting analysis, and quantitative polymerase chain reaction were applied in this study. RESULTS: The mRNA and protein expressions of c-KIT and ANO1 channels were significantly decreased in the colons of DSS-colitis mice. The colonic artificial fecal-pellet transit experiment in vitro was significantly delayed in DSS-colitis mice. The CMMCs and smooth muscle spontaneous contractions were significantly decreased by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), an ANO1 channel blocker, and NG-Nitro-L-arginine methyl ester hydrochloride (L-NAME), an inhibitor of nitric oxide synthase activity, in DSS-colitis mice compared with that of control mice. Intracellular electrical recordings showed that the amplitude of NPPB-induced hyperpolarization was more positive in DSS-colitis mice. The electric field stimulation-elicited nitric-dependent slow inhibitory junctional potentials were also more positive in DSS-colitis mice than those of control mice. CONCLUSION: The results suggest that colonic transit disorder is mediated via downregulation of the nitric oxide/ICC/ANO1 signalling pathway in DSS-colitis mice.
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Animals , Mice , Blotting, Western , Chloride Channels , Colitis , Colon , Dextrans , Down-Regulation , In Vitro Techniques , Interstitial Cells of Cajal , Muscle, Smooth , Myoelectric Complex, Migrating , NG-Nitroarginine Methyl Ester , Nitric Oxide Synthase , Polymerase Chain Reaction , RNA, Messenger , SodiumABSTRACT
Objective: To investigate the effects of three kinds of endometrial preparation (normal hormone replacement, half- and full-dose of long-acting gonadotropin-releasing hormone agonist (GnRH-a) down-regulation combined with hormone replacement) on the pregnancy outcomes in the frozen-thawed embryo transfer (FET) cycle in the patients with endometriosis (E M T), adenomyosis or repeated implantation failure (RIF) for unknown reasons, and to provide a basis for the selection of clinical endometrial preparation method. Methods: A total of 191 patients with EMT, adenomyosis or RIF for unknown reasons underwent FET treatment were selected. The patients were divided into normal hormone replacement group (n=63), half-dose GnRH-a group (n=61) and full-dose GnRH-a group (n=67) according to the endometrial preparation method. The clinical data of patients in each group such as age, body mass index (BMI), duration of infertility, the number of embryo transfer cycles, the number of embryos transferred, the endometrial thickness on the day of conversion and transplantation, the rate of high-quality embryos transferred, the intrauterine clinical pregnancy rate and the embryo implantation rate were analyzed retrospectively and compared. Results: There were no significant differences of the general clinical data of the patients in FET cycles in three groups such as age, BMI, duration of infertility, the number of embryo transfer cycles, the number of embryos transferred, the endometrial thickness on the day of conversion and transplantation, and the rate of high-quality embryos transferred (P
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Objective:To explore the factors affecting the pregnancy outcome of frozen-thawed embryo transfer (FET) in endometriosis (EMT) patients in order to provide reference for the clinical selection of FET strategies. Methods:A total of 329 EMT patients who received blastocyst FET at the Reproductive Medicine Center, Department of Obstetrics & Gynecology, The 900th Hospital of the Joint Logistics Support Force, PLA, from Jan. 2015 to Dec. 2017 were analyzed retrospectively. The patients were divided into three groups according to endometrial preparation protocols, ages, and endometrial thickness on the day of progesterone conversion, respectively. By endometrial preparation protocols, the three groups included gonadotropin-releasing hormone agonist (GnRH-a) down-regulation+ hormone replacement therapy (HRT) group (GnRH-a+HRT group, A1 group, n=138), HRT group (B1 group, n=52), and natural cycle (NC) group (C1 group, n=139). By ages, the three groups included 35 years old group (C2 group, n=59). By endometrial thickness on the day of progesterone conversion, the three groups included 12 mm group (C3 group, n=37). The differences in pregnancy outcomes among EMT patients with blastocyst FET were compared under different grouping factors. Results:The endometrium of A1 group was significantly thicker than that of B1 group (P=0.041), the implantation rate and clinical pregnancy rate of B1 group were significantly higher than those of C1 group (P=0.000, P=0.003). Compared with A1 group, the implantation rate of B1 group was significantly higher (P=0.023), while it was significantly lower in group C1 (P=0.027). The abortion rate of A2 group was significantly higher than that of B2 group (P=0.007). Compared with A3 group, the implantation rate of B3 group was significantly higher (P=0.041), while it was significantly lower in C3 group (P=0.026). Conclusion:HRT endometrial preparation protocol for EMT patients with blastocyst FET can improve the implantation rate and clinical pregnancy rate, and reduce the abortion rate and ectopic pregnancy rate, which may be an economical and efficient endometrial preparation protocol in clinical.
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[Objective] To investigate the effects of different doses of gonadotropin releasing hormone agonist (GnRH-α) on the down-regulation of normal ovarian reserve,and compared the down-regulation level as well as the clinical outcome of in vitro fertilization and embryo transfer (IVF-ET) cycles.[Methods] This RCT study included 63 infertility couples of age＜35 yrs.women with normal ovarian reserve function who were intended to received GnRH-α long protocol treatment.Of the 63 women were randomly divided into three groups according to the dose of triptorelin,21 received daily 0.05 mg short-acting GnRH-α,21 received daily 0.1 mg short-acting GnRH-α,while 21 received reduced-dose depot of 1.25 mg GnRH-αt.[Results] In the three groups,the average duration of down-regulation reached after injection of GnRH-α,the level of LH and E2,the total number of antral follicles,the number of antral follicles of ＜4 mm and 8～9 mm were similar.The serum follicle-stimulating hormone level on the day of gonadotropin initiation were significantly higher in the two short-acting groups compared with the long-acting group [(3.92 ± 1.12) U vs.(3.03 ± 1.14) U vs.(2.05 ± 1.12) U,P＜ 0.001].Four hours after the GnRHa injection,the serum FSH,LH levels were higher in short-acting 0.05 mg group than the short-acting short-acting 0.1 mg group.Both number of days of gonadotropin stimulation and gonadotropin doses were similar in three groups.On the day of hCG administration,the numbers of 14-18mm diameter follicles [(3.91 ±2.12) vs.(5.81 ±3.55) vs.(6.43±3.39),P＜0.001] as well as the proportion of follicles with diameter ≥18 mm/≥10 mm [(33.1％± 13.2％) vs.(24.0％±12.4％) vs.(30.1％±12.2％),P＜0.05],were both statistically significant different in three groups.Although serum LH level on hCG day was significantly increased in 0.05 mg group [(2.47±1.33) U vs.(1.80±0.69) U vs.(1.43±0.53) U,P＜0.05].No premature LH surge and premature ovulation was observed.The number of retrieved oocyteswas significant different [(10.14±4.80) vs.(11.51±2.42) vs.(12.79±2.73),P＜0.05].However,no significant differences was found regard to the number of MII oocytes,and the serum estrogen level per egg was significant higher in 0.05 mg group [(282.33±42.13) U vs.(221.62±32.02) U vs.(200.03±37.89) U,P＜0.001].The live birth rate (LBR) of these three groups in fresh cycles were 61.9％,55.0％,and 50.0％,respectively.The cumulative LBR were 85.7％,76.2％,and 75.0％,respectively.A increased trend was observed in the clinical pregnancy rate,cumulative clinical pregnancy rate and cumulative LBR in 0.05 mg group than the other two groups.[Conclusion] For women with normal ovarian reserve,as the GnRH-α dosage decreased,the down-regulation of pituitary reduced,while serum LH levels on the day of hCG trigger increased.The number of oocytes retrieved was decreased,the proportion of cycles which retrieved ＞ 15 oocytes was also lower.However,the average estrogen level per egg was significant increased,and a better clinical outcome of IVF-ET was received.
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Objective Clinical efficacy was compared among single injections of different doses of long acting gonadotropin releasing hormone agonist (GnRH-a),and daily injections of short-acting GnRH-a in order to evaluate different methods of ovarian stimulation for in vitro fertilization (IVF) cycles.Methods A retrospective study of 214 patients who underwent IVF assisted fertility treatments was conducted.Patients were allocated into four study groups:the short protocol (group A),in which daily injections of 0.1 mg GnRH-a was administered in the mid-luteal phase until the day of human chorionic gonadotropin (hCG) administration (see below);or the long protocol (group B,C & D),in which single injections of 3.75mg,2.0mg,or 0.9mg of long-acting GnRH-a was given in the mid-luteal phase,respectively.Stimulation with gonadotropins (Gn) started when pituitary down-regulation was established.When vaginal ultrasonographic scans showed that at least two follicles had reached 16-20mm in diameter,Gn stimulation was withdrawn,and serum estradiol (E2),progesterone (P),and luteinizing hormone (LH) were determined.Additionally,human chorionic gonadotropin (hCG) was administered that evening.Egg collection was performed 38 hours after hCG injection and the standard IVF procedure was performed.Results There were no statistically significant differences amongst the four groups when measuring serum LH levels,number of oocytes,number of fertilized eggs,number of good quality embryos,and clinical pregnancy rate.The total amount of Gn administered was almost identical when comparing group A and group D,as well as when comparing group B and group C.However,Group A and D required less Gn stimulation to exhibit follicles of 16-20mm in diameter,compared to group B and C (P ＜0.005).Moreover,there was a significant difference in the time required for ovulation induction between group A and group C,where group A had a shorter time to ovulation.The fertilization rate was statistically different between group B and other groups (P ＜ 0.005).Conclusion Through our data analysis,we conclude based on outcome,cost,side-effects,and simplification of treatments,that the 0.9mg long-acting GnRH-a treatment is eminent for ovarian stimulation for IVF.
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Pheochromocytoma is a rare catecholamine secreting neuroendocrine tumor. It is bilateral in 10% of cases. A 24 year lady with bileteral Pheochromocytoma operated for resection under epidural with general anesthesia. Diagnosis was confirmed with 24 hour urinary VMA levels, USG, CT scan and MIBG scan. Tumor resection was incomplete on left side so patient underwent reexploration. Preoperative BP controlled with Phenoxybenzamine and Prazocine. Intra-operative BP fluctuations managed with sodium nitropruside and esmolol. Post operative hypotension was tackled with higher doses of Dopamine and Noradrenline due to down regulation of adrenergic receptors. Patient was discharged on oral steroids after 3 weeks.
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Background Idiopathic orbital inflammatory pseudotumor (IOIP) is a common orbital disease, but its etiology is still unclear,so the effect of glucocorticoid treatment is unsatisfied.Objective This study was to investigate the effects of dexamethasone on orbital fibroblasts from IOIP patients and explore the action machanism.Methods Six pieces of IOIP tissues from 6 IOIP patients and 3 pieces of normal orbital connective tissues from lacrimal gland prolapse patients were obtained during the surgery in Beijing Tongren Hospital from November 2011 to January 2012.The orbital fibroblasts were cultured using explant culture method.The morphology of the cells were observed under the optical microscope,and biomarks of the cells were detected by immunochemistry.The growth and proliferation of the cells were assayed using WST-8.The expression of ICAM-1 in the cells in both the control group and the IOIP group was detected by immunochemistry.The fibroblasts were incubated in 96-well plates, and different concentrations of dexamethasone (0,1 × 10-3 , 1 × 10-4 , 1 × 10-5 and 1 × 10-6 mol/L) were respectively added into the medium for 24,48 and 72 hours,and then the proliferation of the cells was detected by WST-8 assay.The contents of ICAM-1 in different concentrations of dexamethasone groups were assayed by ELISA.Results The characteristics of the cells were similar between the control group and the IOIP group with the spindle shape and long protructions.The cells showed the positive response for vimentin and absent response for desmin, S-100, cytokeratin (CK).Compared with the control group,the growth speed of fibroblasts was fast in the IOIP group.The proliferative values of the cells (absorbancy) were gradually reduced with the increase of dexamethasone concentrations (F ion =36.27,P=0.00) and the lapse of acting time (Ftime =3.69 ,P=0.00).In cultured cells without dexamethasone for 24,48 and 72 hours,the mean expression levels of ICAM-1 were 0.298±0.008,0.312±0.003 and 0.319±0.011, showing a gradually increasing trend.However,the expression of ICAM-1 was gradually reduced with the increases of concentrations and the lapse of acting time of dexamethasone (Fconcentration =75.17,P=0.00;Ftime =3.11,P=0.00).Conclusions Occurrence and development of IOIP is probably associated with the over-expression of ICAM-1 in orbital fibroblasts.Dexamethasone plays anti-inflammation and treating effects on IOIP by down-regulating the expression of ICAM-1 and inhibiting the proliferation of orbital fibroblasts.
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Objective To explore the efficacy of long- and short-acting triptorelin on pituitary down-regulation in long protocol and the pregnancy outcome in vitro fertilization and embryo transfer (IVF-ET) . Methods Three hundred and seventeen patients for IVF-ET were enrolled as study and randomized them into two groups. In group A (n=145), patients received single dose subcutaneous injection of 1.25 mg long-acting diphereline in mid-luteal phase. In group B (n=172), patients received 0.1 mg/d subcutaneous injection of short-acting diphereline in mid-luteal phase for 14-18 days until pituitary suppression were got,and then reduced to 0.05 mg/d until the injection of HCG. Results The dose and the days of gonadotropin administration in group A were bigger and longer than those in group B ( 0.05) . But clinical pregnancy rate of group B had increasing trend. Conclusion Administration of short-acting diphereline has the similar effect with that of long-acting diphereline on pituitary down-regulation in long protocol. Short-acting diphereline requires lower amount of gonadotropin and is more flexible,so it should be recommended.
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MicroRNAs (miRNAs) are small RNAs, 19–23 nucleotides in length, which regulate a variety of cellular processes. Human cytomegalovirus (HCMV) encodes only one intronic miRNA: human cytomegalovirus microRNA UL36 (hcmv-miR-UL36). In this study, we found that over-expression of hcmv-miR-UL36 resulted in a more than threefold increase in HCMV DNA synthesis at 24 h post infection. Fifteen putative targets of hcmv-miR-UL36 were identified using hybrid PCR, one being the HCMV UL138 gene that has previously been identified as a novel latency-associated determinant of HCMV infection. Down-regulation of UL138 expression by hcmv-miR-UL36 was validated using luciferase reporter assays and Western blot analysis in HEK293 cells. In the presence of hcmv-miR-UL36, we observed a 74.6% decrease in luciferase activity and a 46.2% decrease in HCMV UL138 protein expression. Our results indicate that hcmv-miR-UL36 may be a viral miRNA contributing to HCMV replication.
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PURPOSE: Prostaglandin (PG) E2 is known to be closely related to cancer progression and is inactivated by 15-hydroxyprostaglandin dehydrogenase (PGDH). 15-PGDH is shown to have tumor suppressor activity and to be down-regulated in various cancers, including colorectal cancer (CRC). Therefore, we evaluated the expression of 15-PGDH and its prognostic effect in patients with CRC. METHODS: 15-PGDH expression was examined by using immunohistochemistry in 77 patients with CRC. Its prognostic significance was statistically evaluated. RESULTS: Negative 15-PGDH expression was noted in 55.8% of the 77 cases of CRC. 15-PGDH expression showed no correlation with any of the various clinicopathologic parameters. The status of lymph node metastasis, tumor-node-metastasis stages, and pre-operative carcinoembryonic antigen levels showed significant prognostic effect. However, univariate analysis revealed down-regulation of 15-PGDH not to be a predictor of poor survival. The 5-year overall survival rate was 71.7% in the group with positive expression of 15-PGDH and 67.1% in the group with negative expression of 15-PGDH, but this difference was not statistically significant (P = 0.751). CONCLUSION: 15-PGDH was down-regulated in 55.8% of the colorectal cancer patients. However, down-regulation of 15-PGDH showed no prognostic value in patients with CRC. Further larger scale or prospective studies are needed to clarify the prognostic effect of 15-PGDH down-regulation in patients with colorectal cancer.
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Humans , Carcinoembryonic Antigen , Colorectal Neoplasms , Down-Regulation , Hydroxyprostaglandin Dehydrogenases , Immunohistochemistry , Lymph Nodes , Neoplasm Metastasis , Oxidoreductases , Prognosis , Survival RateABSTRACT
Insulin resistance(IR) develops following major injury to the body, including serious trauma, surgical infection and burn. Metabolic disorders due to IR have a serious impact on energy production and attenuate body capacity of anti-infection and anti-shock. Therefore, the research about the mechanism of post-traumatic insulin resistance will be beneficial to improving patients' metabolism situation.
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Objective:To prepare recombinant pCDNA3.1+/Ag85A DNA vaccine encoding tuberculosis Ag85A gene mediated with LipofectamineTM 2000 and to study the effect on T cell subpopulation via oral vaccination.Methods:Recombinant plasmid containing Ag85A using liposome as a vector was constructed and administered to C57BL/6 mice via oral route.Determination of the contents of IFN-r,IL-4 in serum of C57BL/6 mice was performed by double antibody sandwich ELISA.Furthermore,the ability of splenocytes to secret IFN-? and IL-4 was tested by ELISPOT method.When the mice were vaccinated with recombinant eukaryotic expressing vector 5 weeks later,titers of serum antibody against Ag85A were detected by ELISA.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was determined by flow cytometry.Results:Lymphocytes obtained from the spleen of liposomal pcDNA3.1+/Ag85A vaccine-immunized mice exhibited lower IFN-? production and higher IL-4 production than those of pcDNA3.1+ vector immunized mice.The number of spleen MNC secreting IFN-? stimulated by Ag85A protein in vitro was significantly lower than that of plasmid vector group.Liposomal pcDNA3.1+/Ag85A vaccine immunized mice elicited higher Ag85A-specific antibodcy titres.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was decreased.The subset both were shown the profile of Th2 responses.Conclusion:The oral recombinant plasmid pCDNA3.1+/Ag85A mediated with LipofectamineTM 2000 It shows down-regulation effect on the subsets of CD4+ T cells and CD8+ T cells.The regimen has good immunogenecity and could induce Th2 type humoral response in C57BL/6 mice.The immunization suppresses secretion of IFN-?.It can greatly enhance the titres of Ag85A-specific antibodies.LipofectamineTM 2000 can act as an adjuvant through comparation with negative control group.
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Acute tubular necrosis induced by aminoglycoside-antibiotics is followed by a regenerative Process which leads to the restoration of damaged tubules. It is well known that tubular regeneration is mediated by polypeptide growth factors such as epidermal growth factor (EGF) In the absence of nephrotoxic alterations, EGF is immunolocalized in distal tubules, whereas epidermal growth factor receptor(EGFR) immunostaining is localized in proximal tubules. After acute tubular necrosis, the sign of regeneration is accompanied by redistribution of EGF immunoreactivity from distal to proximal tubules and a reduction of total immunoreactive EGF due to a decrease of tissue-bound proEGF. However, it is controversial whether EGFR is down- or up-regulated during this regenerative process. We evaluated the time course of the regenerative response subsequent to tubular damage induced by aminoglycoside, with a particular attention paid to EGFR in order to examine whether it is down- or up-regulated. Female Sprague-Dawley rats were treated during 4 and 8 consecutive days with a daily dose of 80 mg/kg gentamicin i.p. Groups of experimental animals (n=10) were terminated at increasing time intervals (5, 9, 12, 16 days) after cessation of treatment. One hour before sacrifice, each individual received i.p. 200mg/kg Blood for the immunohistochemical demonstration of cell proliferation (S-phase cell). Blood was collected at the time of sacrifice to measure serum creatinine and BUN levels. EGFR immunoreactivity was revealed on paraffin-embedded tissue section by immunohistochemical staining using polyclonal anti-EGFR antibody. Upon immunostaining sections in control kidneys, immunoreactive EGFRs exhibited a quite specific and restricted distribution since they were confined to proximal tubules. But proximal tubules undergoing regenerative repair were characterized by a disappearance of EGFR, which expressed BrdU in immunohistochemical sections for BrdU. Beyond the episode of tubular regeneration, proximal tubules recovered immunoreactive EGFR. The results suggest that the apparent loss of EGFR could be due to a process of receptor down-regulation in proximal tubules displaying evidence of regenerative response.
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Animals , Female , Humans , Rats , Bromodeoxyuridine , Cell Proliferation , Creatinine , Down-Regulation , Epidermal Growth Factor , Gentamicins , Intercellular Signaling Peptides and Proteins , Kidney , Necrosis , Rats, Sprague-Dawley , ErbB Receptors , Regeneration , Withholding TreatmentABSTRACT
The mechanism of ‘down regulation’ of luteinizing hormone receptors was investigated in pseudopregnant rats using a modified radioimmunoassay capable of measuring endogenous tissue-bound hormone. Treatment of pseudopregnant animals with a desensitizing dose (desensitization treatment) of human chorionic gonadotropin resulted in a decrease in receptor concentration. This decrease was prevented if the animals were treated prior to the desensitization treatment with indomethacin, an inhibitor of prostaglandin biosynthesis, suggesting a role for prostaglandins in down regulation. The desensitization treatment resulted in a time-dependent decrease in subsequent responsiveness of the tissue to luteinizing hormone. Basal progesterone production rate was also decreased following desensitization. Total tissue cholesterol was found to be decreased following desensitization treatment, without any change in the ratio of free to esterified cholesterol. Mitochondrial cholesterol was significantly reduced and pregnenolone production by the mitochondria of desensitized corpora lutea was also markedly reduced. However, when cholesterol was added to the mitochondria of desensitized corpora lutea, pregnenolone production was increased, reaching values almost equal to that shown by the control mitochondria. These results show that decrease in the responsiveness following desensitization treatment is due to, besides receptor loss, decrease in tissue cholesterol, in particular mitochondrial cholesterol. The cholesterol side chain cleavage activity, although low, appears to be functionally intact; the low activity could be attributed to low levels of mitochondrial cholesterol.
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The effect of glucocorticoids on the down-regulation of glucocorticoid receptor (GR) mRNA was studied in intact rats. GR mRNA was characterized by Northern blot hybridization and quantitated by dot blot hybridization using a human GR cDNA fragment as a probe. Administration of hydrocortisone in polyvinyl alcohol resulted in a rapid increase in plasma glucocorticoids which maintained at stress levels (20 to 40 ?g/dl) for about 3 d. Hepatic GR mRNA decreased significantly to 73.5?63% of control values 6 h following hydrocortisone treatment, after which the decline of GR mRNA was gradual, reaching a minimum of 44.0?5.0% of control levels 3 d after the treatment The effect of hydrocortisone on the down-regulation of hepatic GR mRNA lasted up to 11 d. In contrast, hydrocortisone treatment had no effect on GR mRNA in rat brain. These results are consistent with the changes in GR in rats as reported previously, except that even though the hepatic cytosol GR decreased markedly, no significant changes in hepatic GR mRNA were found 1 h after hydrocortisone treatment, strongly suggesting that the down-regulation of GR by its ligands in vivo occurs at both transcriptional and posttranscriptional levels and is of tissue-specific fashion.
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Objective:To study the regulatory effects of Ligustrazine Hydrochloride(LHC)on tumor-induced immunosuppression by Colon26 cells in vitro.Methods:Colon26 cells were cultured for 48 h in the presense of LHC and either the cell fraction or the cultural supernatants was collected,with the untreated Colon26 cells as control for the study.The down-regulating effects of LHC on tumor immunosuppressions (including the suppressed NK killing and ConA induced transformation of murine spleen cells detected by MTT,and the reduced expression levels of IL-2R?,CD3?+?+ and CD3?-?+ detected by FCM) were determined.The concentrations of immunosuppressive cytokines,including TGF-?1,VEGF,IL-4,IL-6 and IL-10,in the supernatants were analyzed by quantitative ELISA.The relationship among the down-regulatory effects of LHC on secretion immunosuppressive cytokines and tumor immunosuppressions were evaluated by multiple linear regression analysis.Results:All of the cytokines assayed were found in the supernatant of Colon26 treated without LHC,in which TGF-?1 was the highest,and the significant inhibition of five immune functions mentioned above was showed.To the Colon26 treated by LHC,the concentrations of TGF-?1,IL-6 and IL-10 in the first re-cultured supernatant and its inhibition of five immunol functions decreased greatly.The concentrations of TGF-?1 and IL-6 in the second re-cultured supernatant and its inhibitions of transformation,CD3+?+ and CD3-?+resumed highly.The positive correlations existed between TGF-?1 and inhibition of immunol functions except for transformation,between IL-6 and inhibition of transformation or CD3-?+,between IL-10 and inhibition of NK killing or IL-2R? or CD3+?+,respectively.Conclusion:LHC can exert down-regulate effects on Colon26 secretion of immunosuppressors and its tumor immunosuppression.Reducing tumor immunosuppression of Colon26 through decreasing its secretion of immunosuppressors should be one of anti-tumor mechanisms of LHC.