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OBJECTIVE@#To investigate the expression level of miR-181b in CD19+ B lymphocytes of patients with chronic lymphocytic leukemia (CLL), to analyze the relationship between its expression and the prognosis of CLL patients, and to predict the potential target gene of miR-181b in CLL by using bioinformatics.@*METHODS@#Eight-four patients with CLL treated in People's Hospital of Xinjiang Uygur Autonomous Region from June 2013 to June 2018 were selected. and 20 healthy people were selected as control group. RNA was extracted from CD19+B lymphocytes of peripheral blood by magnetic bead sorting, the expression level of miR-181b was detected, and it's expression differences in different IPI groups were analyzed. The correlation between the expression level of miR-181b and PFS of CLL patients also was analyzed. miR-181b target genes were predicted by online database and literatures, and gene annotation analysis and relevant signal pathway analysis were performed for candidate target genes.@*RESULTS@#The expression level of miR-181b in CLL patients was significantly lower than that in control group (P＜0.01); The expression level of miR-181b in the low-risk group was higher than that in high-risk group and extremely high-risk group (P＜0.05), but there was no statistical difference between low-risk group and medium-risk group (P=1.00). The expression level of miR-181b in medium-risk group was higher than that in high-risk group and extremely high-risk group (P＜0.05), but there was no difference between high-risk group and extremely high-risk group (P=1.00). ROC curve results showed that the area under the curve (AUC) was 0.792 (P＜0.01).When the expression level of miR-181b was at the threshold value of 0.279, it showed a better sensitivity (62.9%) and specificity (91.8%). Survival analysis results suggested that compared with the high expression group, the miR-181b low expression group had poor PFS (log rank: P=0.047). Prediction of miR-181b by using the starBase, targetscan and picTar database and its combination with literature reports indicated that CARD11, ZFP36L1, RUNX1, NR4A3, ATP1B1, PUM1 and PLAG1 related with blood diseases, and up-regulated CARD11 and ZFP36L1 participated in lymphoid tumor formation by promoting cell proliferation and inhibiting cell aging.@*CONCLUSION@#The expression level of miR-181b in CLL group are significantly lower than that in the controls group, and the low expression of miR-181b relates with poor prognosis of CLL patients. Through bioinformatics prediction and combined with literature reports, it is speculated that CARD11 and ZFP36L1 as target genes of miR-181b may be participated in the occurrence and development of CLL. Further experiments are needed to verify this result.
Subject(s)
Humans , Apoptosis Regulatory Proteins , Cell Proliferation , Leukemia, Lymphocytic, Chronic, B-Cell , Genetics , MicroRNAs , PrognosisABSTRACT
Objective MicroRNA-145 (miR-145) is underexpressed in breast cancer. The study aimed to explore the regulatory effect of miR-145 on breast cancer MCF-7 cells by investigating the association of miR-145 with ADAM17 and EGFR. Methods The MCF-7 breast cancer cells were divided into three groups: the transfection group (transfected with microRNA-145 mimics), the control group (without transfection) and the nonsense sequence group (transfected with nonsense microRNA). MTT, transwell and real-time quantitative fluorescence polymerase chain reaction(qPCR) were respectively used to detect the proliferative capacity, invasive ability and expression of MCF-7 breast cancer cells after the transfection of miR-145 in three groups. ADAM17 and EGFR mRNA and protein levels in three groups of breast cancer MCF-7 cells were detected by qPCR and western blot. Results The results of qPCR showed that the relative expression of miR-145 was significantly higher in transfection group (13964.33±1265.30) than those in control group (1.00±0.05) and nonsense sequence group (1.03±0.15) and the difference was statistically significant (P<0.01); the expression of ADAM17 mRNA in transfection group (1.71±0.08) was significantly higher than that in control group (1.00±0.07) and the difference was statistically significant (P<0.01). Compared with the nonsense sequences at 24 h, 48 h, and 72 h, the inhibition rate of MCF-7 in transfection group was significantly increased (P<0.01). The results of transwell invasion showed that the number of transmembrane cells in transfection group [(56.20±2.17)/field] was significantly lower than those in control group [(92.80±3.90)/field] and nonsense sequence group [(91.80±4.97)/field of view ] (P < 0.01). Western blot results showed that the protein content of ADAM17 and EGFR in transfection group was significantly lower than those in the control group and the nonsense sequence group (P<0.01). Conclusion MiR-145 inhibits the proliferation and invasion of breast cancer MCF-7 cell line by acting on the ADAM17-EGFR signaling pathway.
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<p><b>OBJECTIVE</b>To detect whether the murine T-cell lymphoma cell line EL4 could be infected by murine cytomegalovirus (MCMV), and to observe the morphological changes and apoptosis of El4 cells before and after infection.</p><p><b>METHODS</b>EL4 cells were infected with MCMV smith strain with 1, 10 and 100 multiplicity of infection (MOI) respectively. The morphology of the cells was observed by light microscopy and Wright's-Giemsa staining. The survival rate was calculated by trypan blue staining. RT-PCR-based assay was used to detect the copy number of MCMV-DNA in the infected ELA cells. Flow cytometry was used to detect apoptosis. RT-qPCR was used to detect the mRNA expression levels of P53, P21, cFlip and Caspase 8. The protein expression levels of Caspase8, P53, BAX, BCL-2 and Cleaved Caspase3 proteins were detected by Western blot.</p><p><b>RESULTS</b>After Wright-Giemsa staining, it was found that the infected EL4 cells displayed larger volume, irregular nuclei and the folded twist under light microscopy. Compared with the normal control group, the survival rate of EL4 cells decreased, and the apoptosis rate statistically significantly (P<0.05) increased with the increasing MOI and the infected time in each group. While, the level of apoptosis protein P53, BAX/BCL-2, Cleaved-caspase3 and Caspase8 were up-regulated. And the survival rate, apoptosis rate and the apoptosis protein level of infected EL4 cells with MOI=10 were the most obvious at the 5day. Compared with MCMV infection group (MOI=60), the content of MCMV DNA in EL4 cells was decreased in MCMV+GGV group ［MOI=60, GCV 25 (g/ml)］, and the cell apoptosis rate and apoptosis protein expression of P53, Caspase8, BAX/BCL-2 were decreased (P<0.05).</p><p><b>CONCLUSION</b>Murine T-cell lymphoma cell line EL4 can be infected by MCMV and displayes an obvious apoptosis phenomenon. MCMV may up-regulate the expression levels of apoptosis protein P53, BAX-BCL-2, Cleaved-caspase3 and Caspase8 in EL4 cells. The drug ganciclovir reduces the copy mumber of MCMV DNA in infected EL4 cells and inhibited the killing effect of MCMV on EL4 cells.</p>
Subject(s)
Animals , Mice , Apoptosis , Caspase 8 , Ganciclovir , Lymphoma, T-Cell , MuromegalovirusABSTRACT
Objective To explore bone formation markers in dexamethasone intervention osteocalcin (OC),bone alkaline phosphatase (BAKP),and type Ⅰ original amino terminal propcptide (PINP) relationship with bone longitudinal growth.Methods The selected thirty-three 4-week-old male SpragueDawley (SD) rats were randomly divided into two groups:the dexamethasone group (n =18) and the control group (n =15).The rats in the dexamethasone group received dexamethasone (200 μg/100 g) by intraperitoneal injection for 10 days.The rats in the control group received matching volume sodium chloride solution.All rats were weighed everyday.The rats were killed by using 10％ chloral hydration at 8 AM of 11 th day.The length of tibiae was measured.The proximal tibiae were excised,fixed and decaleified.Mter paraffin embedded,sections in 5 μm thick were cut.The growth plate sections were stained by haematoxylin-eosin (HE) histochemistry method.Total height of growth plate was measured.The rats decaptitating and the blood were collected.Serum was separated and stored in-80 ℃ refrigerator for analysis.Enzyme -linked immuno sorbent assay (ELISA) method was used to detect the rat OC,BAKP and PINP values.Results The length of growth plate and tibiae of dexamethasone group were significantly decreased contrast the control group:the length of growth plate (P =0.001),and the length of tibiae (P =0.000).There were no significant differences between two groups of the value of OC,BAKP and PINP:OC (P =0.056),BAKP (P=0.122),and PINP (P =0.169).There was positive correlation between the serum OC and the length of tibiae (r =0.454,P =0.08) in control group,the PINP and OC (r =0.521,P =0.026) in dexamethasone group.Conclusions Glucocorticoid inhibit the longitudinal bone growth,to the osteoblasts (OC,BAKP and PINP) of growing rats is not obvious.
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Objective To investigate whether cell lines expressing apolipoprotein E-enhanced green fluorescence protein (apoE-EGFP) can support assembly of infectious hepatitis C virus particles. Methods apoE stably down-regulated Huh7. 5. 1 cell lines (sh-apoE cell lines) were established by shRNA gene silencing technique, and cell lines expressing apoE-EGFP fusion protein (apoE-EGFP cell lines) were established. The culture supernatants of wild-type Huh7. 5. 1 cells, control cells (sh-NT cells), sh-spoE cells and apoE-EGFP cells transfected with HCV RNA were collected and the HCV titer of supernatants was determined by TCID50. The interaction of apoE with HCV structure protein E2 was examined by immunofluorescence and confocal microscopy, and the expression of apoE-EGFP on the surface of HCV particles purified by FLAG-specific affinity gll was analyzed by Western blotting analysis. Results The apoE-EGFP fusion protein was highly expressed in sh-apoE cell lines. The infectivity of HCV from apoE-EGFP cell culture supernatant was not significantly different with those of HCV from Huh7. 5. 1 and sh-NT cells. apoE-EGFP infused protein had fluorescent co-location with HCV structural protein E2, and was detected on the surface of HCV particles purified by FLAG-specific affinity gll. Conclusion The apoE-EGFP fusion protein is an important component of HCV particles and apoE-EGFP cell lines can support the assembly of HCV particles.
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<p><b>OBJECTIVE</b>To explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.</p><p><b>METHODS</b>Using combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.</p><p><b>RESULTS</b>Sertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).</p><p><b>CONCLUSION</b>The proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.</p>
Subject(s)
Animals , Male , Rats , Cells, Cultured , Glial Cell Line-Derived Neurotrophic Factor , Metabolism , Rats, Wistar , Sertoli Cells , Cell Biology , Metabolism , Temperature , Testis , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To observe improved effects of Jingjin acupuncture on fine activity of hemiplegic hand in recovery period of stroke.</p><p><b>METHODS</b>Fifty cases were randomly divided into an observation group and a control group, 25 cases in each one. Regular western medicine treatment, rehabilitation training and regular acupuncture (in which Shuigou (GV 26), Baihui (GV 20), Neiguan (PC 6), etc. were selected) were applied in both groups. Additionally, muscles in palm side of affected hand, dorsal metacarpophalangeal joints and proximal interphalangeal joints were treated with acupuncture in the observation group, once every other day and electroacupuncture was applied when arrival of qi was acquired. Baxie (EX-UE 9) in the affected hand were needled in the control group, and electroacupuncture was added when arrival of qi was acquired. Ten days of treatment was considered a treatment course, and after two courses Lindmark score, Brunnstrom movement function grade, joint range of hand and Barthel index (BI) were observed in two groups.</p><p><b>RESULTS</b>Compared before the treatment, the Lindmark score in two groups were both improved after the treatment (both P < 0.01). Compared with the control group, the motor coordination ability, sensory function and total score of Lindmark in observation group were obviously improved (differences before and after treatment: 8.24 +/- 3.07 vs 6.84 +/- 2.43, 3.52 +/- 2.33 vs 2.16 +/- 2.12, 11.76 +/- 3.55 vs 9.00 +/- 3.62, all P < 0.05). The Brunnstrom movement function grade was significantly improved in both groups after treatment (both P < 0.01), which was more obvious in the observation group (P < 0.05). The joint range of hemiplegic hand was improved in both groups after treatment (both P < 0.01), which was more obvious in the observation group [differences before and after treatment: (25.35 +/- 10.91) degrees vs (18.65 +/- 7.86) degrees, p < 0.05]. The score of BI was also significantly improved after treatment in two groups (both P < 0.01).</p><p><b>CONCLUSION</b>The Jingjin acupuncture could effectively improve fine activity of hemiplegic hand in recovery period of stroke prove daily life ability.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Acupuncture Therapy , Hand , Hemiplegia , Therapeutics , Movement , Recovery of Function , StrokeABSTRACT
OBJECTIVE: To evaluate the bioequivalence of domestic sirolimus capsules and commercially available sirolimus oral solution in healthy Chinese volunteers. METHODS: Twenty-two healthy Chinese male volunteers were enrolled and orally administered 5 mg sirolimus capsules or oral solution randomly. The concentrations of sirolimus in whole blood were determined by LC-MS/MS. RESULTS: The pharmacokinetic parameters of sirolimus capsules and oral solution were as following: ρmax(26.62 ± 9.97) and (25.28 ± 5.98) ng · mL-1; tmax(2.27 ± 0.55) and. (1.91 ± 2.33) h; t1/2 (73.07 ± 13.81) and (65.49 ± 17.00) h; AUC0-t (372.3 ± 146.2) and (368.2 ± 275.0) ng middot; h middot; mL-1, AUC0-∞ (466.8 ± 194.3) and (442.3 ± 324.4) ng middot; h middot; mL-1, respectively. The relative bioavailability F0-t was (112.4 ± 34.61)% and F0-∞ was (117.2 ± 39.93)%. Two-sided t-test of ρmax, AUC0-t and AUC0-∞ indicated that the preparations were equivalent, and non-parametric test showed that sirolimus capsules had larger tmax than the reference preparation with a significant difference (P < 0.05). CONCLUSION: The domestic sirolimus capsule is bioequivalent to the marketed sirolimus oral solution with significantly different tmax. Copyright 2012 by the Chinese Pharmaceutical Association.
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<p><b>OBJECTIVE</b>To observe autoantibodies production against AT(1)-receptors and alpha(1)-adrenergic receptors and association to risk factors, such as sex, age, family history, course of hypertension and other cardiovascular diseases in hypertensive patients.</p><p><b>METHODS</b>A total of 690 patients with essential hypertension admitted to our hospital were selected and autoantibodies against AT(1)-receptors and alpha(1)-adrenergic receptors were detected by ELISA. Multiple logistic regression analysis was performed based on obtained data.</p><p><b>RESULTS</b>Positive rates for antibody against AT(1)-receptors and alpha(1)-adrenergic receptors were 47.1% (325/690) and 36.4% (251/690) respectively in this group of patients. Duration of hypertension history was significantly longer in the antibody against AT(1)-receptors and alpha(1)-adrenergic receptors positive groups [(9.3 +/- 11.0) year, (9.9 +/- 11.1) year] compared to the negative groups [(7.3 +/- 9.3) year, (7.2 +/- 9.5) year, all P < 0.01]. The ratio of family history with hypertension was also significantly higher in antibody positive groups than negative ones (47.69% vs 39.18%, P < 0.01). Regression analysis demonstrated that 5 risk factors were related to positive production of autoantibody against AT(1)-receptors including female gender, age, family history, duration of hypertension history and diabetes. However, just age, family history, duration of hypertension history were main factors responsible to the production of autoantibody against alpha(1)-adrenergic receptors (all P < 0.05).</p><p><b>CONCLUSION</b>The environmental and genetic factors contributed to the autoantibody production in patients with essential hypertension.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Autoantibodies , Blood , Hypertension , Blood , Allergy and Immunology , Receptor, Angiotensin, Type 1 , Allergy and Immunology , Receptors, Adrenergic, alpha , Allergy and Immunology , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To observe the association between positive autoantibodies against AT(1)-receptor and cardiac remodeling in primary hypertensive patients.</p><p><b>METHODS</b>Echocardiography was performed and serum autoantibodies against AT(1)-receptor were detected by enzyme linked immunosorbent assay (ELISA) in 592 patients with primary hypertension. The differences on blood pressure level, course of hypertension, vasoactive substance and echocardiography parameters between the positive group and negative group were compared. Factors related to left ventricular enlargement were analyzed by multiple logistic regressions.</p><p><b>RESULTS</b>The positive percentage of autoantibodies against AT(1)-receptor was 38.0% (225/592). End-diastolic right atrial and left ventricular diameters in positive group were significantly larger than that in negative group (P = 0.049 and P = 0.044, respectively). Regression analysis demonstrated that positive autoantibodies against AT(1)-receptor, male gender, diastolic blood pressure and course of hypertension were related to left ventricular enlargement (all P < 0.05).</p><p><b>CONCLUSION</b>The autoantibodies against AT(1)-receptor is associated with left ventricular and right atrial enlargement in hypertensive patients.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Autoantibodies , Blood , Heart Ventricles , Hypertension , Blood , Allergy and Immunology , Hypertrophy, Left Ventricular , Receptor, Angiotensin, Type 1 , Allergy and Immunology , Ventricular RemodelingABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of autoantibodies against alpha(-) adrenergic receptor on cardiac remodeling in patients with hypertension.</p><p><b>METHODS</b>Five hundred and fifty three patients with hypertension in our hospital were selected. The autoantibodies against alpha(1) adrenergic receptor in sera of donor were detected by ELISA, and the results of echocardiography were recorded. By multiple logistic regressions, the risk factors were analyzed on left ventricular enlargement of hypertension.</p><p><b>RESULTS</b>The percentage of autoantibodies against alpha(1) adrenergic receptor positive was 32.3% (179/553). There were significant difference between the positive group and negative group on the ratio of left atrial enlargement (53.6%, 44.3%, respectively; P < 0.05) and left ventricular enlargement (12.8%, 6.1%, respectively; P < 0.01). The result of regression analysis demonstrated that 4 risk factors were related to left ventricular enlargement, including male, course of disease, heart rate (HR) and autoantibodies against alpha(1) adrenergic receptor in the serum (all P < 0.05).</p><p><b>CONCLUSIONS</b>The autoantibodies against alpha(1) adrenergic receptor have a relationship with left ventricular enlargement of hypertension. Patients with the activity of autoantibodies against alpha(1) adrenergic might contribute to predict cardiac remodeling.</p>