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2.
Article in English | WPRIM | ID: wpr-901411

ABSTRACT

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

3.
Article in English | WPRIM | ID: wpr-893707

ABSTRACT

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⁺ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

4.
Article in Chinese | WPRIM | ID: wpr-751578

ABSTRACT

Cardioembolic stroke (CES) is the most severe type of ischemic stroke, accounting for 14% to 30% of all ischemic strokes, tends to early recurrence, and has a high long-term recurrence rate and mortality. Therefore, early diagnosis of CES in patients with ischemic stroke is of great significance. Some studies have shown that brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) are associated with CES. This article reviews the roles of BNP and NT-proBNP in early diagnosis and recurrence risk prediction of CES.

5.
Article in Chinese | WPRIM | ID: wpr-699756

ABSTRACT

Objective To investigate the role of epigenetic regulations of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) in the development of diabetic retinopathy and the metabolic memory phenomenon after hyperglycemia was terminated.Methods Diabetic rat model was established by intraperitoneal injection of streptozotocin (STZ).Sixty diabetic rats were randomly divided into 3 groups,poor glycemic control group rats were maintained in poor glycemic control for 4 months;semi glycemic control group rats were maintained in poor glycemic control for 2 months,followed by good glycemic control for 2 additional months;good glycemic control group rats were maintained in good glycemic control for 4 months.Twenty normal rats served as control group.The mRNA expression of PGC-1α and superoxide dismutase 2 (SOD2) of retina were measured by real-time PCR;the expression of PGC-1α and manganese superoxide dismutase (MnSOD) protein were measured by Western blot;the situation of DNA methylation in the promotor region of PPARGC1A was measured by bisulfite sequencing.Results The body-weight in the control group was significantly higher than that in the poor glycemic control group,semi glycemic control group and good glycemic control group (all at P =0.000).The blood glucose value in the poor glycemic control group was significantly higher than that in the control group (P =0.000).The expression levels of PGC-1 α mRNA were significantly lower and the expression levels of SOD2 mRNA were significantly higher in the good glycemic control group,semi glycemic control group and poor glycemic control group than those in the control group (all at P<0.05).The expression levels of PGC-1α and SOD2 mRNA were significantly different between the good glycemic control group and poor glycemic control group (both at P<0.05).Compared with the control group,the expression levels of PGC-1α and MnSOD protein were decreased in the diabetic model groups,with significant differences between them (all at P<0.05).The expression level of PGC-1 α protein was significantly higher in the good glycemic control group than that in the poor glycemic control group (P<0.05).Diabetes increased DNA methylation in the promotor region of PPARGC1A gene of retina.The DNA methylation level was significantly higher in the poor glycemic control group and semi glycemic control group than that in the control group (P =0.008,0.031).No statistical difference was found between the poor glycemic control group and semi glycemic control group (P > 0.05).Conclusions The expressions of PGC-1o mRNA and protein and MnSOD protein in the retina of STZ induced diabetic rats are decreased,the expression of SOD2 mRNA is increased,the expression changes have metabolic memory characteristics.Increased DNA methylation in the promotor region of PPARGC1A when exposed to high glucose may have a role in the regulation of PGC-1 α expression and metabolic memory.

6.
Article in Chinese | WPRIM | ID: wpr-615369

ABSTRACT

This study aimed to use the sika deer as a model to study the influence of IGF-1 on the expression of Col Ⅰ in antler chondrocytes.The chondrocytes were separated from sika deer antlers,cultured and were treated with recombinant human IGF-1 protein (rIGF-1),both rIGF-1 and PQ401,and transfected with IGF-1 over-expression plasmid or IGF-1 siRNA,respectively.The expression of Col Ⅰ which,a well-known marker for chondrocytes dedifferentiation,was detected by real-time PCR.The results showed that administration of rIGF-1 to antler chondroctyes resulted in an obvious decrease of Col Ⅰ mRNA levels,while PQ401 pretreatment could dramatically attenuate the effects of rIGF-1 on the expression of Col Ⅰ mRNA.After transfection with IGF-1 over-expression plasmid,the expression of Col Ⅰ mRNA was obviously reduced in antler chondrocytes compared with control.Conversely,knockdown IGF1 with specific siRNA could increase the expression of Col Ⅰ in antler chondrocytes.These results indicate that IGF-1 may play an important role in process of antler chondrocyte dedifferentiation.

7.
The Journal of Practical Medicine ; (24): 2431-2435, 2017.
Article in Chinese | WPRIM | ID: wpr-611784

ABSTRACT

Objective To investigate the effect of melatonin on oxidative stress and inflammatory reaction in patients with moderate to severe chronic obstructive pulmonary disease and to explore its mechanisms. Methods 42 patients with moderate to severity chronic obstructive pulmonary disease in stable stage were random-ly divided into melatonin group and control group,and 21 patients in each group treated with melatonin(3 mg/d) or placebo for 3 months respectively. The plasma levels of 8- isoprotane,IL-8,TNF-α,h-CRP pulmonary func-tion,six minutes walking test and MRC dyspnea score before treatment,2 months and 3 months after the treatment were analyzed. Results After 2 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(10.40 ± 5.4 vs. 16.92 ± 4.33,P<0.05),and the concentration of IL-8 (6.88 ± 2.37 vs. 11.33 ± 3.39,P < 0.05). After 3 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(9.40 ± 4.0 vs. 17.92 ± 3.33,P < 0.01),and IL-8 (5.67 ± 3.22 vs. 9.31 ± 3.23,P < 0.05). Compared with before treatment,melatonin could significantly de-creased the concentration of 8-isoprotane(9.40 ± 4.0 vs. 20.40 ± 8.4,P<0.01 )and IL-8(5.67 ± 3.22 vs. 12.33 ± 3.88,P<0.05)after 3 months. Meanwhile,the concentration of the TNF-α(25.83 ± 9.18 vs. 35.83 ± 12.18,P<0.05)and hypersensitive C(1.76 ± 1.18 vs. 3.09 ± 1.79,P < 0.05)reactive protein in the melatonin group was greatly lower than the placebo group. After 3 months,compared to the placebo group,MRC dyspnea score of pa-tients in the group of melatonin was improved significantly(1.56 ± 1.38 vs. 2.09 ± 1.16,P<0.05 ),and lung func-tion and six minutes walk test showed no significant difference between patients in the two groups. Conclusions Exogenous melatonin administration can decrease the concentration of 8-isoprotane,IL-8,TNF-αand h-CRP in the blood of patients with moderate to severe COPD ,and improve the MRC dyspnea score. Melatonin has a significant effect on reducing oxidative stress and inhibiting inflammatory reaction in patients with moderate and severe stage stable COPD,which demonstrates its potential therapeutic value with broad clinical application prospects.

8.
Chinese Journal of Oncology ; (12): 245-249, 2017.
Article in Chinese | WPRIM | ID: wpr-808553

ABSTRACT

Objective@#To investigate the effects of simvastatin on proliferation, invasion and radiosensitivity of mouse Lewis lung cancer cell line in vitro.@*Methods@#The inhibitory effects of simvastatin on proliferation of Lewis lung cancer cells were detected by MTT assay. Matrigel invasion and migration assay was used to determine the invasion and motility ability of the Lewis cells. P38 activity was measured by p38 activity detection kit, and the expressions of p-p38, MKP-1, RhoA and MMP-2 were analyzed by Western blot. Lung cancer xenograft model was established in C57BL/6 mice. The mice were randomly divided into control group, simvastatin group, radiotherapy alone group and combined treatment group. The mice were killed 27 days after inoculation. The tumor mass, volume and lung metastatic nodules in the mice were compared.@*Results@#The cell proliferation rates of 0 μmol/L, 10 μmol/L, 20 μmol/L and 30 μmol/L simvastatin groups were 100%, (87.0±9.0)%, (76.5±8.1)% and (67.0±7.3)%, respectively (P<0.05). Invasive cell numbers of the above groups were 298±30, 251±26, 207±20 and 132±19 per field, respectively (P<0.05). The intracellular p38 activities were 100%, (83.1±8.8)%, (70.2±8.2)% and (59.0±6.4)%, respectively. The relative expressions of p-p38 were 100%, (76.2±6.7)%, (56.4±5.4)% and (36.5±3.2)%, respectively. The expressions of RhoA were 100%, (80.1±5.3)%, (55.3±6.2)% and (38.6±4.8)%, respectively. The expressions of MMP-2 were 100%, (89.6±8.6)%, (51.9±4.7)% and (42.7±3.1)%, respectively, while the expressions of MKP-1 were 100%, (136.5±12.2)%, (168.8±15.3)% and (187.7±13.4)%, respectively (all P<0.05). Lung metastatic nodules and mass in the control, simvastatin, radiotherapy group and combined treatment groups were 6.24±1.09, 3.07±0.71 g, 5.09±1.16, 2.43±0.53 g, 3.12±0.68, 1.96±0.62 g and 2.65±0.38, 1.12±0.43 g, respectively (all P<0.05). The tumor inhibition rates were 39.0%, 48.1% and 26.5%, respectively, in the radiotherapy alone, combined treatment and simvastatin groups (all P<0.05).@*Conclusions@#Simvastatin inhibits the proliferation of Lewis cell line by inhibiting the activity of p38 and expression of p-p38. Meanwhile, simvastatin reduces the invasion and motility of Lewis cell line through down-regulating the expression of RhoA and MMP-2. When combined with radiotherapy, simvastatin can inhibit tumor growth and metastasis, and improve the treatment efficacy of radiotherapy synergistically.

9.
Article in Chinese | WPRIM | ID: wpr-504809

ABSTRACT

Objective:To construct the pleiotrophin (PTN )overexpression vector,and to explore the effect of PTN on the decidualization of uterine stromal cells in the mice.Methods:The specific primers containing restriction enzyme cutting sites were designed according to the PTN gene sequences published in GenBank for PCR amplification.The amplified fragment of PTN was recovered from the agarose gel and cloned into the pGEM-T vector.The pGEMT-PTN was cut by double enzyme digestion and ligated into pcDNA3.1 (+)to construct the PTN overexpression plasmid.After transfection with PTN overexpression plasmid,the expression levels of PTN mRNA in the uterine stromal cells and the expression levels of decidualization markers Prl8a2 and Prl3c1 were detected by qRT-PCR method.The uterine stromal cells transfected with pcDNA3.1 (+)empty vector were used as control group. Results:The results of identification by double enzyme digestion indicated that the bands of PTN overexpression plasmid were consistent with those of the target gene,and the clone sequencing results suggested that it had 100% homology with mouse PTN gene sequence published in GenBank.Compared with control group, the expression levels of PTN,Prl8a2 and Prl3c1 mRNA in mouse uterine stromal cells in PTN overexpression group were significantly increased (P < 0.05).Conclusion:PTN overexpression could increase the expression levels of decidualization markers in mouse uterine stromal cells,indicating that PTN might play an enhancement effect during uterine decidualization in the mice.

10.
Article in Chinese | WPRIM | ID: wpr-636004

ABSTRACT

Background Diabetic retinopathy (DR) is one of the leading causes that result in adult irreversible blindness in many countries.Recent researches suggest that neurodegeneration is an important component of DR.To realize the disease process of retinal neutron is very important for prevention and treatment on DR.Objective This study was to investigate the change of retinal nerve fiber layer thickness in patients with type 2 diabetes mellitus.Methods Ninety-six eyes of 48 patients with type 2 diabetes mellitus were enrolled in Peking Union Medical College Hospital.The patients were assigned into non-diabetic retinopathy (NDR) group,background diabetic retinopathy(BDR) group,proliferative diabetic retinopathy (PDR) group and panretinal photocoagulation (PRP) group based on the fundus finding and fundus fluorescein angiography(FFA),and 24 normal subjects with matched age were included as control group.RNFL thickness was measured by GDxVCC system,including temporal,superior,nasal,inferior,total,(TSNIT) average,superior average,inferior average,TSNIT standard deviation and nerve fiber indication.The datas of the RNFL thickness were analyzed and comparison among different groups by one-way analysis of variance and Student Newman Keuls test.Results The TSNIT averages of the NDR group,BDR group,PDR group and PRP group were(56.54±5.28),(56.92±6.49),(53.04±6.14) and(53.17±9.30) μm,respectively,while that of the control group was (59.04±4.37) μm.The TSNIT average,superior average,inferior average,TSNIT standard deviation of the PDR group and PRP group compared with control group were significantly decreased,and the nerve fiber indication of the PDR group and PRP group was significantly increased (P =0.002,0.000,0.002,0.000,0.001 ;P =0.002,0.000,0.001,0.000,0.000).Compared with the control group,the TSNIT average,superior average,inferior average,TSNIT standard deviation were insignificantly decreased,and the nerve fiber indication was insignificantly increased in the NDR group and BDR group (P =0.187,0.235,0.333,0.106,0.202 ;P=0.262,0.063,0.072,0.098,0.062).Conclusions The decline of the RNFL thickness appears prior to DR findings.The RNFL thinning of PDR and PRP patients suggests the degeneration of neurons and atrophy of axonal.The neurodegeneration is an important component of DR.

11.
Article in English | WPRIM | ID: wpr-233162

ABSTRACT

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.


Subject(s)
Animals , Bronchi , Metabolism , Calcium , Metabolism , Calcium Channels , Calcium Signaling , Physiology , Cells, Cultured , Male , Membrane Glycoproteins , Metabolism , Myocytes, Smooth Muscle , Metabolism , ORAI1 Protein , Protein Kinase C-delta , Metabolism , Rats , Rats, Sprague-Dawley , Stromal Interaction Molecule 1
12.
Chinese Medical Journal ; (24): 1134-1138, 2011.
Article in English | WPRIM | ID: wpr-239878

ABSTRACT

<p><b>BACKGROUND</b>Cytomegalovirus (CMV) retinitis is the most severe intraocular complication that results in total retinal destruction and loss of visual acuity in patients with acquired immunodeficiency syndrome (AIDS). This study aimed to investigate the fundus characteristics, systemic manifestations and therapeutic outcomes of CMV retinitis associated with AIDS.</p><p><b>METHODS</b>It was a retrospective case series. CMV retinitis was present in 39 eyes (25 patients). Best corrected visual acuities, anterior segment, fundus features, fundus fluorescence angiography (FFA) and CD4(+) T-lymphocyte counts of the patients with CMV retinitis associated with AIDS were analyzed. Intravitreal injections of ganciclovir (400 µg) were performed in 4 eyes (2 patients).</p><p><b>RESULTS</b>Retinal vasculitis, dense, full-thickness, yellow-white lesions along vascular distribution with irregular granules at the border, and hemorrhage on the retinal surface were present in 28 eyes. The vitreous was clear or mildly opaque. Late stage of the retinopathy was demonstrated in 8 eyes characterized as atrophic retina, sclerotic and attenuated vessels, retinal pigment epithelium (RPE) atrophy, and optic nerve atrophy. Retinal detachment was found in 3 eyes. The average CD4(+) T-lymphocyte count in peripheral blood of the patients with CMV retinitis was (30.6 ± 25.3) × 10(6)/L (range, (0 - 85) × 10(6)/L). After intravitreal injections of ganciclovir, visual acuity was improved and fundus lesions regressed.</p><p><b>CONCLUSIONS</b>CMV retinitis is the most severe and the most common intraocular complication in patients with AIDS. For the patients with yellow-white retinal lesions, hemorrhage and retinal vasculitis without clear cause, human immunodeficiency virus (HIV) serology should be performed. Routine eye examination is also indicated in HIV positive patients.</p>


Subject(s)
Acquired Immunodeficiency Syndrome , Allergy and Immunology , Metabolism , Adult , Antiviral Agents , Pharmacology , CD4-Positive T-Lymphocytes , Metabolism , Cytomegalovirus Retinitis , Drug Therapy , Allergy and Immunology , Metabolism , Female , Fluorescein Angiography , Ganciclovir , Pharmacology , Humans , Male , Middle Aged , Retrospective Studies , Young Adult
13.
Article in Chinese | WPRIM | ID: wpr-384899

ABSTRACT

Stroke is one of the severe perioperative complications.The incidence of perioperative stroke has great difference because of the surgical site,surgical complexity,and different basic diseases of patients.The induced anesthesia and the intraoperative management also have an important influence on the onset of stroke.The mortality of perioperative stroke is as high as 26%.Neturologists must guide the assessment of the potential risks for perioperative stroke,and carefully balance the risk-benefit ratio when making each decision.This article reviews the advances in research on the risk factors for perioperative stroke,prevention,and treatment.

14.
Article in Chinese | WPRIM | ID: wpr-229938

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the incidence of myopic retinopathy and its risk factors.</p><p><b>METHODS</b>The fundus of 1449 patients (2879 eyes) with myopia were retrospectively examined. The clinical relationship between myopic retinopathy and diopter, age, and sex was analyzed.</p><p><b>RESULTS</b>Myopic retinopathy was detected in 413 eyes (14.35%). Posterior pole retinal lesions were detected in 22 eyes (0.76%). Peripheral retinal lesions were found in 396 eyes (13.75%). According to their diopters, the myopic patients were divided into four groups: low, medium, high and super high myopia The incidence of peripheral retinal lesions was 4.18%, 8.72%, 19.18%, and 37.44% in these four groups, which significantly different (chi2 = 178.594, P<0.001). By age these patients were divided into three groups: I group, age <25; II group, age 25-34; III group, age >34. The incidences of peripheral retinal lesions in these three groups were 8.11%, 15.34%, and 24.59%, which were significantly different (chi2 = 76.090, P<0.001). The incidence of retinal lesion in male and female was 9.32% and 16.07%, respectively, which was significantly different (chi2 = 24.886, P<0.001). Posteriorpole retinal lesions were only detected in the highly or super highly myopic patients, all of them were more than 25 years. The incidence of posteriorpole retinal lesions in the highly and super highly myopia group was 0.86% and 6.67% respectively, which was significantly different (chi2 = 31.898, P<0.001). The incidence of posteriorpole retinal lesions in group II and group III was 0.55% and 3.55% respectively, which was significantly different (chi2 = 22.523, P<0.001).</p><p><b>CONCLUSIONS</b>The prevalence of retinal lesions in myopic patients is higher than that of emmetropia. The incidence of peripheral retinal lesions increases in patients with deeper diopters. Posterior pole retinal lesions usually occur in the myopic patients whose age are more than 25 years and diopter more than - 6.00 D. Careful examination of fundus is essential for early detection and timely treatment.</p>


Subject(s)
Adult , Female , Humans , Male , Myopia , Retina , Pathology , Retinal Diseases , Pathology , Retrospective Studies , Young Adult
15.
Article in Chinese | WPRIM | ID: wpr-534029

ABSTRACT

AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

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