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1.
China Journal of Chinese Materia Medica ; (24): 2360-2367, 2023.
Article in Chinese | WPRIM | ID: wpr-981312

ABSTRACT

This study explored the effect and underlying mechanism of Stellera chamaejasme extract(SCE) on multidrug resistance of breast cancer. The chemotherapy-sensitive breast cancer cell line MCF-7 and adriamycin(ADR)-resistant cell line MCF-7/ADR were used as experimental subjects. MTT assay was used to detect cell proliferation activity. Pi staining was used to detect the cell cycle. 4',6-Diamidino-2-phenylindole, dihydrochloride(DAPI) staining and flow cytometry were used to detect apoptosis. Dansylcadaverine(MDC) staining and GFP-LC3B-Mcherry adenovirus transfection were used to detect autophagy. The protein expression of Bcl-2, Bax, caspase-9, caspase-3, LC3B, p62, and Beclin-1 was detected by Western blot. The results showed that SCE could significantly inhibit the proliferation of both sensitive and resistant breast cancer cell lines. The drug resistance factor was 0.53, which was significantly lower than 59 of ADR. Meanwhile, the proportion of sensitive/resistant cells in the G_0/G_1 phase increased significantly after SCE treatment. In addition, DAPI staining showed that a series of apoptosis phenomena such as nuclear pyknosis, staining deepening, and nuclear fragmentation appeared in sensitive/resistant cell lines after SCE administration. Moreover, the results of flow cytometry double staining showed that the proportion of apoptotic cells in sensitive/resistant cell lines increased significantly after SCE administration. Besides, Western blot showed that the protein expression levels of caspase-3, caspase-9, and Bcl-2 significantly decreased and the expression level of Bax protein significantly increased in both breast cancer cell lines after SCE administration. Furthermore, SCE could also increase the positive fluorescent spots after MDC staining and yellow fluorescent spots after GFP-LC3B-mcherry transfection, and up-regulate the expression levels of autophagy-related proteins LC3B-Ⅱ, p62, and Beclin-1 in breast cancer cells. In summary, SCE may play the role of anti-multidrug resistance by blocking the cell cycle of breast cancer multidrug-resistant cells, blocking autophagy flow, and ultimately interfering with the apoptosis resistance of drug-resistant cells.


Subject(s)
Humans , Female , Breast Neoplasms/metabolism , MCF-7 Cells , Caspase 3/metabolism , Caspase 9/metabolism , Beclin-1/pharmacology , Apoptosis , Proto-Oncogene Proteins c-bcl-2/metabolism , Cell Line, Tumor , Drug Resistance, Neoplasm , Cell Proliferation
2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 124-131, 2021.
Article in Chinese | WPRIM | ID: wpr-905997

ABSTRACT

Objective:To establish an ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) for determining the plasma concentrations of 10 active ingredients in Wujiwan at different time points after oral administration, and to compare the pharmacokinetic characteristics between normal rats and rats with chronic visceral hypersensitive irritable bowel syndrome (CVH-IBS). Method:CVH-IBS rat model was prepared by the neonatal rat colon percutaneous transluminal coronary angioplasty (PTCA) balloon stimulation method. After intragastric administration of Wujiwan (0.245 g·kg<sup>-1</sup>), blood was collected from the jugular vein at different time points, and the plasma concentrations of 10 active ingredients (berberine hydrochloride, palmatine hydrochloride, coptisine hydrochloride, jatrorrhizine hydrochloride, epiberberine, dihydroberberine, evodiamine, evodine, paeoniflorin, albiflorin) in Wujiwan was detected simultaneously by UPLC-MS/MS, the pharmacokinetic parameters of each component in normal rats and CVH-IBS rats were calculated. Result:The established UPLC-MS/MS could sensitively and accurately detect the plasma concentrations of 10 active ingredients of Wujiwan in rats. Compared with the normal group, the absorption rates of these 10 active ingredients of Wujiwan in the blood of CVH-IBS rats all decreased to a certain extent, and the peak time (<italic>t</italic><sub>max</sub>) was prolonged. Among them, the <italic>t</italic><sub>max</sub> of berberine hydrochloride and jatrorrhizine hydrochloride were significantly prolonged from 54 minute and 39 minute to 90 minute, respectively (<italic>P</italic><0.05, <italic>P</italic><0.01). Area under the plasma concentration-time curve (AUC<sub>0-</sub><italic><sub>t</sub></italic>) of each component increased, and evodiamine and paeoniflorin were significantly different (<italic>P</italic><0.05,<italic> P</italic><0.01). The clearance rates (CL/<italic>F</italic>) of these 10 active ingredients were all decreased, among which berberine hydrochloride, palmatine hydrochloride and evodiamine had significant differences (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:There are significant differences in the pharmacokinetic behavior of the active ingredients in Wujiwan between normal rats and CVH-IBS rats, which may be related to the destruction of microstructure of intestinal epithelial cells and the change of activity of liver enzymes under the pathological state of IBS.

3.
China Journal of Chinese Materia Medica ; (24): 3402-3409, 2021.
Article in Chinese | WPRIM | ID: wpr-887991

ABSTRACT

This study aimed to investigate the effect and the possible mechanism of Shenlian( SL) extract on tumor necrosis factor-α( TNF-α)-induced ECV304 injury. After the establishment of TNF-α-induced ECV304 cells injure model,MTT assay was used to detect cell viability and the level of reactive oxygen species( ROS) was measured by flow cytometry. The contents of superoxide dismutase( SOD),malondialdehyde( MDA),nitric oxide( NO),endothelin-1( ET-1) and interleukin-1β( IL-1β) in the supernatant were detected by biochemical method and enzyme linked immunosorbent assay( ELISA). The expression levels of apoptosis-related proteins B-lymphoma-2 gene( Bcl-2),Bcl-2 associated X protein( Bax),caspase-3,caspase-9 and nuclear factor E2 associated factor2( Nrf2)/Kelch like epichlorohydrin associated protein-1( Keap1) signaling pathway related proteins Nrf2,Keap1,quinone oxidoreductase( NQO1) and heme oxygenase 1( HO-1) were detected by Western blot. The results showed that 50 μg·L-1 TNF-α significantly damaged ECV304 cells,induced the impairment of cell viability( P<0. 01),the increase of ROS production,the decrease of SOD activity,and the increase of MDA,NO,ET-1 and IL-1β( P<0. 01),meanwhile,it caused the up-regulation of Keap1,caspase-9 and Bax protein expression,and down-regulation of NQO1 and Bcl-2 protein expression( P<0. 05) compared with the control group.Compared with the model group,SL extract reduced the damage of ECV304 cells induced by TNF-α,improved cell viability,reduced ROS production,increased SOD activity and decreased MDA,NO,ET-1,IL-1β content( P<0. 01 or P<0. 05). In addition,SL extract also down-regulated the protein expression levels of Keap1,caspase-3,caspase-9 and Bax,and increased the protein expressions of Nrf2,NQO1,HO-1 and Bcl-2( P<0. 01 or P<0. 05). The above results indicate that SL extract can provide protective effect on ECV304 cells injury induced by TNF-α,alleviate oxidative stress injury,inflammation and apoptosis,and its mechanism may be related to regulating Nrf2/Keap1 signaling pathway.


Subject(s)
Apoptosis , Kelch-Like ECH-Associated Protein 1/metabolism , NF-E2-Related Factor 2/metabolism , Oxidative Stress , Plant Extracts , Signal Transduction , Tumor Necrosis Factor-alpha/genetics
4.
Journal of Experimental Hematology ; (6): 259-264, 2021.
Article in Chinese | WPRIM | ID: wpr-880064

ABSTRACT

OBJECTIVE@#To investigate the difference in the therapeutic effect of plasma exchange and continuous renal replacement therapy (PE+CRRT) combined with chemotherapy in the treatment of children with severe Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) and non-EBV-HLH.@*METHODS@#The clinical data of 21 cases of all children with severe HLH treated by PE+CRRT combined with chemotherapy from January 2017 to January 2020 were collected and retrospectively analyzed. According to the presence of EBV infection, the children were divided into EBV@*RESULTS@#Among the 21 children, 14 were divided into the EBV@*CONCLUSION@#PE+CRRT combined with chemotherapy can reduce serum ferritin quickly, then improve organ function, and increase the overall survival rate of severe HLH, and it is a good effect on children with severe EBV-HLH and non-EBV-HLH.


Subject(s)
Child , Humans , Continuous Renal Replacement Therapy , Epstein-Barr Virus Infections/complications , Herpesvirus 4, Human , Lymphohistiocytosis, Hemophagocytic , Plasma Exchange , Retrospective Studies
5.
Chinese Journal of Contemporary Pediatrics ; (12): 744-748, 2020.
Article in Chinese | WPRIM | ID: wpr-828673

ABSTRACT

OBJECTIVE@#To study the clinical features of children with severe adenovirus pneumonia (SAP) and hemophagocytic syndrome (HPS).@*METHODS@#A retrospective analysis was performed from the chart review data of 30 children with SAP and HPS who were admitted from January 2014 to June 2019. According to the prognosis, the children were divided into a good prognosis group (n=18) and a poor prognosis group (n=12).@*RESULTS@#Among the 30 children with SAP and HPS, the ratio of male to female was 2:1. The median age of onset was 1 year and 3 months (range 3 months to 5 years), and the mean course of fever was 19±7 d. Of the 30 children, 28 (93%) experienced disease onset in January to June. High-throughput gene detection of serum pathogens showed that 16 (53%) children were positive for human adenovirus type 7 (HAdV-7), and the other 14 (47%) children were positive for HAdV antigen based on immunofluorescence assay for throat swab, with unknown type. Of all 30 children, 29 (97%) had respiratory complications, 24 (80%) had cardiovascular complications, 16 (53%) had gastrointestinal complications, and 9 (30%) had toxic encephalopathy. Eighteen children (60%) improved or recovered and 12 (40%) did not recover (3 died). Compared with the good prognosis group, the poor prognosis group had a significantly longer course from onset to diagnosis of HPS (P<0.05), significantly higher levels of fibrinogen and tumor necrosis factor-α (P<0.05), and a significantly lower level of interferon-γ (P<0.05). The mean follow-up time was 6±2 months; 11 (41%) children recovered, 1 (4%) experienced recurrence of HPS, and 15 (56%) had the sequela of post-infectious bronchiolitis obliterans (PIBO).@*CONCLUSIONS@#HPS may be observed in children with SAP, and PIBO is the most common sequela of SAP.


Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Adenoviridae , Adenoviridae Infections , Lymphohistiocytosis, Hemophagocytic , Pneumonia, Viral , Retrospective Studies
6.
Journal of Experimental Hematology ; (6): 29-33, 2020.
Article in Chinese | WPRIM | ID: wpr-781492

ABSTRACT

OBJECTIVE@#To explore whether BAX plays a role in the development of Philadelphia chromosome-positive leukemia and related mechanisms.@*METHODS@#Target-gene knockout mice were used as bone marrow cell donors. Retrovirus over-expressing BCR-ABL were packaged. BCR-ABL-induced B-ALL mouse model was established through donor's B cells transfected by the retrovirus and the B cells over-expressing BCR-ABL were given to the receptor mice by tail vein injection. Western blot was used to detect the protein express and flow cytometry was used to analyze the B cell subpopulations in BAX and WT mouse bone marrows. Kaplan-Meier analysis was used to estimate the survival of diseased mice.@*RESULTS@#BAX deletion caused faster development of BCR-ABL-induced leukemia in vitro and in vivo. BCR-ABL increased BCL-2 expression and enhanced BCL-2/BAX heterodimer formation.@*CONCLUSION@#The BAX deletion can accelerate the disease progression of BCR-ABL induced B-ALL.

7.
Journal of Experimental Hematology ; (6): 539-544, 2019.
Article in Chinese | WPRIM | ID: wpr-771923

ABSTRACT

OBJECTIVE@#To explore whether the high risk factors possibly leading to hypercoagulative status and thrombosis exist in Thalassemia patients of Guangxi region through detecting plasma tissne factor-bearing microparticles (TFMP), procoagulatima activity, coagulation and anticoagulation function, fibrinolytic function, endothelial function and platelet count.@*METHODS@#The TFMP procoagulation activity was detected by chromogenic saubstract method, the levels of tissue factors (TF), tissue factor pathway inhibitor(TFPI), protein C (PC), protein S (PS), antithrombin Ⅲ(AT-Ⅲ), tissue plasminogen activator (tPA), thrombin-activated fibrinolysis inhibitor (TAFI), soluble E-selectin (sE-sel), intercellular adhesion molecule-1 (ICAM-1) and thrombomodulin (TM) were detected by ELISA in thalassemia group (n=71) and control group (n=20 heathy persons).@*RESULTS@#Compared with control group, the AT-Ⅲ level decreased in β-thalastemia major group (TM) (P<0.05), the AT-Ⅲ level in TM group independeutly posstiody correlated with plt count (r=0.37, P<0.05); the levels of TF and sICAM in α-thalassenia intermediate group (TA) significantly decteased (P<0.05), the procoagulatim activity of TFMP in β-thalassemia intermediate group (TI) increased sngnificantly (P<0.05), moreover positively corretated with AT-Ⅲ level (r=0.77, P<0.05). The TF and sICAM-1 levels in normal liver functim group of Thalassemia patients were lower tham those in control group (P<0.01 and P<0.05, respectively), the TFMP activity between normal and abnormal liver function was significantly different (P<0.05), while there were no significant difference in other correspoding indexes beween thalassemia group and control group as well as between each thalassemia groups.@*CONCLUSION@#The damage of liver function and reduction of anticoagylation substances exist in patients with β-thalassenia major in Guangxi region, the procoagulation activity of plasma TFMP in patients with β-thalassemia intermedia abnormally increases. All the above-mentioned factors may increase the risk of high coagulation status or thrombosis is thalassemia patients, the decrease of TF and SICAM-1 levels in patients with α-thalassemia intermedia may be factor against thrombosis.


Subject(s)
Humans , Anticoagulants , Antithrombin III , China , Thalassemia , Thromboplastin , Tissue Plasminogen Activator
8.
Journal of Experimental Hematology ; (6): 134-140, 2019.
Article in Chinese | WPRIM | ID: wpr-774346

ABSTRACT

OBJECTIVE@#To detect the expression of miR-99a-5p in myelodysplastic syndrome (MDS), to predict the target genes and to analyze its function by using bioinformatics.@*METHODS@#The expression levels of bone marrow miR-99a-5p in MDS patients were detected by qRT-PCR, and the correlation of miR-99a-5p expression with clinical pathological characteristics, percentage of marrow blasts , chromosome karyotype and peripheral blood hemogram were analyzed. The target genes of miR-99a-5p were predicted by Targetscan, Miranda and Microcosm, and the intersection of the predicted results of 3 softwares was used as a potential target gene for miR-99a-5p.@*RESULTS@#The expression level of bone marrow miR-99a-5p in MDS patients was significantly higher than that of healthy controls (P<0.01); According to the prognostic score of IPSS, MDS patients were divided into relatively low risk group (including low risk group and intermediate risk group 1) and relatively high risk group (including intermediate risk group 2 and high risk group). Analysis showed that the expression level of bone marrow miR-99a-5p in relatively low risk group was 2.40 times higher than that in healthy control group (P<0.01), the expression level of bone marrow miR-99a-5p in relatively high risk group was 6.66 times higher than that in healthy control group (P<0.01), the expression level of miR-99a-5p in relatively high risk group was 2.80 times higher than that in the relatively low risk group ( P<0.01) ; the expression level of bone marrow miR-99a-5p in t-MDS group was 6.35 times higher than that in healthy control group (P<0.001). There was a significant positive correlation between the expression level of miR-99a-5p and the percentage of marrow blasts (P<0.01), but the expression of miR-99a-5p did not correlate with chromosome karyotype and peripheral blood hemogram; In this study it was obtained that ST5 might participate in pathological mechanism of MDS by bioinformatic analyses and references.@*CONCLUSION@#The expression levels of miR-99a-5p is up-regulated in MDS patients, and its expression showed a rising tendency which may be involved in the pathogenesis of MDS by regulating target gene ST5.


Subject(s)
Humans , Bone Marrow , Computational Biology , Karyotyping , MicroRNAs , Myelodysplastic Syndromes
9.
Journal of Experimental Hematology ; (6): 1749-1753, 2019.
Article in Chinese | WPRIM | ID: wpr-781402

ABSTRACT

OBJECTIVE@#To investigate the effect of BAX gene deletion on the sensitivity of BCR-ABL-induced B-ALL cells of mice to imatinib and the related mechanism.@*METHODS@#The target gene-knock out (BAX) mice were used as bone marrow cell donors; the wild type bone marrow cells(B6BM) and BAX bone marrow cells(B6BM-BAX) of mice were transfected by using reverse transcription virus, then the BCR-ABL transfected B6BM cells and B6BM-BAX cells were treated with imatinib at different concentration (0,0.5, 1.0 and 2.0 μmol/L) for 48 hours. The number of viable cells was detected by trypan blue, the flow cytometry was used to detect the cell apoptosis, the Western blot was used to detect the changes of BAX, Caspase expression.@*RESULTS@#In BCR-ABL transfected bone marrow cells treated with imatinib, the numbers of viable cells of BAX deletion group was significantly higher than that of wild type groups with statristcal difference(P<0.05), and effect- and dose-dependency(r=-0.9533 for BAX deletion group, and r=-0.9812 for wild type group). The flow cytometry showed that the cell apoptosis in BAX deletion group signifincantly decreased, compared with wild type group(P<0.05). The Western blot showed that the expression of apoptotic protein Caspase 3 in BAX deletion group was significantly higher than that in wild type group(P<0.05).@*CONCLUSION@#BAX deletion can reduce the sensitivity of BCR-ABL-induced B-ALL cells to imatinib.


Subject(s)
Animals , Mice , Apoptosis , Drug Resistance, Neoplasm , Fusion Proteins, bcr-abl , Gene Deletion , Imatinib Mesylate , Piperazines , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma , Genetics , bcl-2-Associated X Protein
10.
Chinese Traditional Patent Medicine ; (12): 379-382, 2018.
Article in Chinese | WPRIM | ID: wpr-710199

ABSTRACT

AIM To study the chemical constituents from Zanthoxylum bungeanum Dahongpao.METHODS The 95% ethanol extract from Z.bungeanum Dahongpao was purified by silica column and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as β-sitosterol (1),hydroxy-β-sanshool (2),daucosterol (3),quercetin-3-O-galactoside (4),3-methoxyquercetin (5),succinic acid (6),mannitol (7),arbutin (8),7-O-α-Dglucosyl-3,8-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)-5-methoxy-4H-1-benzopyran-4-one (9).CONCLUSION Compounds 5 and 7 are isolated from genus Zanthoxylum for the first time,compounds 1,3-9 are first isolated from this plant.

11.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 684-690, 2018.
Article in Chinese | WPRIM | ID: wpr-737255

ABSTRACT

U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries.These fractures are highly unstable and frequently cause neurological deficits.The majority of surgeons have limited experience in management of U-shaped sacral fractures.No standard treatment protocol for U-shaped sacral fractures has been available till now.This study aimed to examine the management of U-shaped sacral fractures and the early outcomes.Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed.Demographics,fracture classification,mechanism of injury and operative treatment and deformity angle were assessed.All the patients were treated with lumbopelvic fixation or (and) sacral decompression.EQ-5d score was applied to evaluate the patients' quality of life.Of the 15 consecutive patients with U-shaped sacral fracture,the mean age was 28.8 years (range:15-55 years) at the time of injury.There were 6 females and 9 males.The mean followup time was 22.7 months (range:9-47 months) and mean full weight-bearing time was 9.9 weeks (range:8-14 weeks).Ten patients received lumbopelvic fixation and sacral decompression,one lombosacral fixation,and 4 merely sacral decompression due to delayed diagnosis or surgery.The post-operation deformity angle (mean 27.87°,and range:8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67;range:15-90) with no significance difference noted.At the latest follow-up,all patients obtained neurological recovery with different extents.Visual analogue score (VAS) was reduced from preoperative 7.07 (range:5-9) to postoperetive 1.93 (range:1-3).All patients could walk without any aid after treatment.Eight patients were able to care for themselves and undertook some daily activities.Five patients had returned to work full time.In conclusion,lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed.Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery.Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.

12.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 684-690, 2018.
Article in Chinese | WPRIM | ID: wpr-735787

ABSTRACT

U-shaped sacral fractures are rare and often difficult to diagnose primarily due to the difficulty in obtaining adequate imaging and the severe associated injuries.These fractures are highly unstable and frequently cause neurological deficits.The majority of surgeons have limited experience in management of U-shaped sacral fractures.No standard treatment protocol for U-shaped sacral fractures has been available till now.This study aimed to examine the management of U-shaped sacral fractures and the early outcomes.Clinical data of 15 consecutive patients with U-shaped sacral fracture who were admitted to our trauma center between 2009 and 2014 were retrospectively analyzed.Demographics,fracture classification,mechanism of injury and operative treatment and deformity angle were assessed.All the patients were treated with lumbopelvic fixation or (and) sacral decompression.EQ-5d score was applied to evaluate the patients' quality of life.Of the 15 consecutive patients with U-shaped sacral fracture,the mean age was 28.8 years (range:15-55 years) at the time of injury.There were 6 females and 9 males.The mean followup time was 22.7 months (range:9-47 months) and mean full weight-bearing time was 9.9 weeks (range:8-14 weeks).Ten patients received lumbopelvic fixation and sacral decompression,one lombosacral fixation,and 4 merely sacral decompression due to delayed diagnosis or surgery.The post-operation deformity angle (mean 27.87°,and range:8°-90°) of the sacrum was smaller than that pre-operation (mean 35.67;range:15-90) with no significance difference noted.At the latest follow-up,all patients obtained neurological recovery with different extents.Visual analogue score (VAS) was reduced from preoperative 7.07 (range:5-9) to postoperetive 1.93 (range:1-3).All patients could walk without any aid after treatment.Eight patients were able to care for themselves and undertook some daily activities.Five patients had returned to work full time.In conclusion,lumbopelvic fixation is an effective method for stabilization of U-shaped sacral fractures with fewer complications developed.Effective reduction and firm fixation are the prerequisite of early mobilization and neurological recovery.Sacral decompression effectively promotes neurological recovery even in patients with old U-shaped sacral fractures.

13.
Journal of Clinical Pediatrics ; (12): 486-488, 2016.
Article in Chinese | WPRIM | ID: wpr-496373

ABSTRACT

Objective To explore the function of autonomic nerves system in children with congenital heart disease (CHD) combined with paroxysmal supraventricular tachycardia (PSVT). Methods Fifty children having PSVT and no CHD (PSVT group), 30 children with both PSVT and CHD (CHD group), and 50 cases of healthy children (control group) were selected. The difference of 24-hour heart rate variability (HRV) among three groups was analyzed retrospectively. Results There were statistical differences in the long-range time domain HRV indexes, including SDNN, SDANN, SDNN Index, pNN 50 , and RMSSD among three groups (F=80.32-?263.18, P all??0.05). The SDNN, SDANN, SDNN Index, pNN50, and RMSSD were signiifcantly decreased in CHD group compared with PSVT group (P all?

14.
Acta Physiologica Sinica ; (6): 50-56, 2016.
Article in English | WPRIM | ID: wpr-331683

ABSTRACT

Myocytes in the pulmonary veins (PV) play a pivotal role in the development of paroxysmal atrial fibrillation (AF). It is therefore important to understand physiological characteristics of these cells. Studies on these cells are, however, markedly impeded by the fact that single PV myocytes are very difficult to obtain due to lack of effective isolation methods. In this study, we described a novel PV myocyte isolation method. The key aspect of this method is to establish a combination of retrograde heart perfusion (via the aorta) and anterograde PV perfusion (via the pulmonary artery). With this simultaneous perfusion method, a better perfusion of the PV myocytes can be obtained. As results, the output and viability of single myocytes isolated by simultaneous heart and PV perfusion method were increased compared with those in conventional retrograde heart perfusion method.


Subject(s)
Animals , Rabbits , Atrial Fibrillation , Cell Separation , Heart , Muscle Cells , Perfusion , Pulmonary Veins
15.
Journal of Experimental Hematology ; (6): 1476-1483, 2016.
Article in Chinese | WPRIM | ID: wpr-332666

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of miR-550a-5p in bone marrow of patients with myelodysplastic syndrome (MDS), and to predict its target genes and function by bioinformatics analyses, so as to provide the evidence to furthre explore the role of miR-550a-5p and its target genes in pathogenesis of MDS.</p><p><b>METHODS</b>Real-time PCR was used to detect the expression of miR-550a-5p in 54 MDS patients, 16 acute myeloid leukemia transformed from MDS (sfAML) and 19 healthy controls, and the correlation between the expression of miR-550a-5p and clinical pathologic characteristics of MDS, including chromosome, percentage of marrow blasts, absolute neutrophil count, platelet count and hemoglobin levels were analyzed. The sequence of miR-550 was searched in miRBase database. Target genes of miR-550a-5p were predicted by Microcosm,Miranda and Targetscan, and the predective results were collected, then the enrichment analyses of target gene function(GO) and signalling pathway(pathway of miR-550a-5p) were carried out by using gene ontology darabase and KEGG database.</p><p><b>RESULTS</b>The expression of miR-550a-5p in bone marrow of all MDS patients was higher than that in controls: the expression level of miR-550a-5p in low risk MDS and middl risk 1 MDS was 1.7 times of controls (P=1.23×10); the expression of miR-550a-5p in midde risk 2 MDS and high risk MDS was 1.9 times of controls (P=1.20×10); the expression of miR-550a-5p in tAML was 2.0 times of controls (P=5.61×10). The miR-550a-5p expression level was up-regulated gradually with the enhancement of disease risk of MDS, but there was no correlation between the expression level of miR-550a-5p and clinical pathologic characteristics of MDS(chromosome: Normal: 1.11±0.19, Abnormal:1.26±0.15, P>0.05; Percentage of Marrow Blasts: r=0.29,P=0.07; absolute neutrophil count: r=-0.02,P=0.89; hemoglobin level: r=0.09,P=0.57; platelet count: r=0.25,P=0.08). The sequence of miR-550 was conservative among different species, and the prediced results indicated that there were 19 target genes in intersection. The functions of target genes were enriched in regulation of stress-activated cascade, MAPK pathway, regulation of muscle organ development, regulation of protein homodimerization activity and other biological processes; they participated in some molecular functions including enzyme activity, combination processes of some molecules as protein, cAMP and domain existed in cell junction, synapse, coated vesicle, dendrite and other cellular components. Two of them-PDLIM2 and PSME1 were selected which might play a role in pathologic mechanism of MDS regulated by miR-550a-5p.</p><p><b>CONCLUSION</b>The expression of miR-550a-5p in bone marrow of MDS patients increases specifically, and miR-550a-5p may play a role in the pathogenesis of MDS through regulation of target genes, PDLIM2 and PSME1.</p>

16.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 541-547, 2016.
Article in English | WPRIM | ID: wpr-285232

ABSTRACT

This study examined the clinical outcomes of one-stage surgical treatment for patients with spinal tuberculosis via a posterior-only approach. Twenty-four patients with thoracic or lumbar spinal tuberculosis whose lesions were confined to adjacent segments were admitted to our hospital and treated. The American Spinal Injury Association (ASIA) impairment scale was used to assess the neurological function. All patients were treated with one-stage surgical treatment via a posterior-only approach. The clinical efficacy was evaluated by the Japanese Orthopaedic Association (JOA) scores and oswestry disability index (ODI) of nerve function. Patients were evaluated preoperatively and postoperatively by measurement of spinal deformity using Cobb angle and radiological examination. All the patients were followed up for 13 to 27 months. They had significantly postoperative improvement in JOA score, ODI and ASIA classification scores. The kyphotic angles were significantly corrected and maintained at the final follow-up. Bone fusion was achieved within 4-12 months. It was concluded that one-stage surgical treatment via a posterior-only approach is effective and feasible for the treatment of spinal tuberculosis.


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bone Transplantation , Debridement , Fracture Fixation, Internal , Plastic Surgery Procedures , Spinal Fusion , Tuberculosis, Spinal , Diagnostic Imaging , General Surgery
17.
Chinese journal of integrative medicine ; (12): 944-948, 2015.
Article in English | WPRIM | ID: wpr-287153

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of gastrodin on isolated thoracic aorta rings of rats and to investigate the potential mechanism.</p><p><b>METHODS</b>A perfusion model of isolated thoracic aorta rings of rats was applied. The effect of cumulative gastrodin (5, 50, 100,150, 200, and 250 μmol/L) on endothelium-intact aorta rings was investigated. The same procedure was applied to observe the effect of gastrodin on endothelium-intact/denuded aorta rings pre-contracted with 10(-6) mol/L phenylephrine hydrochloride (PE). The aorta rings incubated by 200 mmol/L gastrodin in the Ca(2+)-free (K-H) solution was contracted by using PE. The effect of 200 mmol/L gastrodin on endothelium-denuded aorta rings pre-contracted with 60 mmol/L KCl was also observed.</p><p><b>RESULTS</b>Compared with the denuded gastrodin group, the intact gastrodin group could significantly relax the PE-contracted aorta rings (P<0.01). In Ca(2+)-free (K-H) solution KHS, the PE-induced contraction rate of aorta rings pre-incubated by gastrodin was 6.5%±0.7%, which was significantly less than the control group (11.8%±0.9%,P<0.01). However, after 3 mmol/L CaCl2 was added, the Ca(2+)-induced contraction in the gastrodin group (51.7%±2.4%) was similar to that in the control group (49.8%±2.8%). The contractile rate of rings in the KCl-contracted gastrodin group (96.3%±0.6%) was not significantly different from that in the control group (96.8%±1.2%).</p><p><b>CONCLUSIONS</b>Gastrodin has the effect of vasorelaxation on isolated thoracic aorta rings of rats. The mechanism of the vasorelaxation of gastrodin may mainly work through the inhibition of inositol 1, 4, 5-trisphosphosphate receptor on the sarcoplasmic reticulum of the arterial smooth muscle, which leads to the reduction of the Ca(2+) released from the sarcoplasmic reticulum.</p>


Subject(s)
Animals , Female , Male , Rats , Aorta, Thoracic , Physiology , Benzyl Alcohols , Pharmacology , Calcium , Metabolism , Endothelium, Vascular , Physiology , Glucosides , Pharmacology , In Vitro Techniques , Phenylephrine , Pharmacology , Rats, Wistar , Vasodilation
18.
Chinese journal of integrative medicine ; (12): 188-195, 2015.
Article in English | WPRIM | ID: wpr-310913

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of Dan-gua Fang on adenosine 5'-monophosphate (AMP) activated protein kinase (AMPK) α expression in liver and subsequent improvement of glucose and lipid metabolism.</p><p><b>METHODS</b>Forty 13-week-old diabetic Goto-Kakizaki (GK) rats were randomly divided into model, Dan-gua Fang, metformin and simvastatin groups (n=10 for each), and fed high-fat diet ad libitum. Ten Wistar rats were used as normal group and fed normal diet. After 24 weeks, liver expression of AMPKα mRNA was assessed by real-time PCR. AMPKα and phospho-AMPKα protein expression in liver was evaluated by Western blot. Liver histomorphology was carried out after hematoxylin-eosin staining, and blood glucose (BG), glycosylated hemoglobin A1c (HbA1c), food intake and body weight recorded.</p><p><b>RESULTS</b>Similar AMPKα mRNA levels were found in the Dan-gua Fang group and normal group, slightly higher than the values obtained for the remaining groups (P<0.05). AMPKα protein expression in the Dan-gua Fang group animals was similar to other diabetic rats, whereas phospho-AMPKα (Thr-172) protein levels were markedly higher than in the metformin group and simvastatin group (P<0.05), respectively. However, phosphor-AMPKα/AMPKα ratios were similar in all groups. Dan-gua Fang reduced fasting blood glucose with similar strength to metformin, and was superior in reducing cholesterol, triglycerides, high-density lipoprotein cholesterol as well as improving low-density lipoprotein cholesterol in comparison with simvastatin and metformin. Dan-gua Fang decreases plasma alanine aminotransferase (ALT) significantly.</p><p><b>CONCLUSION</b>Dan-gua Fang, while treating phlegm-stasis, could decrease BG and lipid in type 2 diabetic GK rats fed with high-fat diet, and effectively protect liver histomorphology and function. This may be partly explained by increased AMPK expression in liver. Therefore, Dan-gua Fang might be an ideal drug for comprehensive intervention for glucose and lipid metabolism disorders in type 2 diabetes mellitus.</p>


Subject(s)
Animals , Male , AMP-Activated Protein Kinases , Genetics , Metabolism , Blood Glucose , Metabolism , Body Weight , Diabetes Mellitus, Experimental , Blood , Drug Therapy , Metabolism , Drugs, Chinese Herbal , Therapeutic Uses , Feeding Behavior , Glycolipids , Metabolism , Liver , Pathology , Phosphorylation , RNA, Messenger , Genetics , Metabolism , Rats, Wistar , Real-Time Polymerase Chain Reaction , Time Factors
19.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1086-1095, 2014.
Article in Chinese | WPRIM | ID: wpr-294341

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of Dangua Recipe (DGR) on glycolipid metabolism, vascular cell adhesion molecule-1 (VCAM-1) and its mRNA expression level of transgenic Apo E(-/-) mouse with spontaneous atherosclerosis, thus revealing its partial mechanism for curing diabetes mellitus (DM) with angiopathy.</p><p><b>METHODS</b>Diabetic model was prepared by peritoneally injecting streptozotocin (STZ) to Apo E(-/-) mouse. Totally 32 modeled mice were stratified by body weight, and then divided into 4 groups referring to blood glucose levels from low to high by random digit table, i.e., the model group (MOD, fed with sterile water, at the daily dose of 15 mL/kg), the DGR group (fed with DGR at the daily dose of 15 mL/kg), the combination group (COM, fed with DGR at the daily dose of 15 mL/kg and pioglitazone at the daily dose of 4.3 mg/kg), and the pioglitazone group (PIO, at the daily dose of 4.3 mg/kg), 8 in each group. Another 8 normal glucose C57 mouse of the same age and strain were recruited as the control group. All interventions lasted for 12 weeks by gastrogavage. The fasting blood glucose (FBG), body weight, food intake, water intake, skin temperature, the length of tail, and the degree of fatty liver were monitored. The hemoglobin A1c (HbA1c), total cholesterol (TC), and LDL-C were determined. Endothelin-1 (ET-1) was determined by radioimmunoassay. Nitrogen monoxidum (NO) was determined by nitrate reductase. The kidney tissue VCAM-1 level was analyzed with ELISA. The expression of VCAM-1 mRNA in the kidney tissue was detected with real time quantitative PCR.</p><p><b>RESULTS</b>Compared with the control group, the body weight and food intake decreased, water intake increased in all the other model groups (P < 0.05). Besides, the curve of blood glucose was higher in all the other model groups than in the control group (P < 0.01). Compared with the model group, the body weight increased; levels of HbAlc, TC, LDL-C, ET-1, and VCAM-1 were significantly lower; and skin temperature was higher in the DGR group (P < 0.05, P < 0.01). Compared with the PIO group, body weight, the increment of body weight, FBG, TC, and LDL-C were lower (P < 0.05, P < 0.01); food intake and water intake increased more and the tail length was longer in the DRG group (P < 0.01). There was no statistical difference in the level of NO among groups. The degree of fatty liver in the model group was significantly severer than that in the control group (P < 0.05). It was obviously alleviated in the DGR group (P < 0.05) when compared with the model group and the PIO group (P < 0.05, P < 0.01). But it was severer in the PIO group than in the model group (P < 0.01). The degree of fatty liver in the combination group ranged between that of the DGR group and the PIO group (P < 0.05). The level of VCAM-1 mRNA expression was significantly lower in the DGR group than in the model group, the PIO group, and the combination group (P < 0.05).</p><p><b>CONCLUSIONS</b>DGR had effect in lowering blood glucose and blood lipids, and fighting against fatty liver of transgenic Apo E(-/-) mouse with spontaneous atherosclerosis. DGR played an effective role in preventing and treating DM with angiopathy by comprehensively regulating glycolipid metabolism and promoting the vascular function.</p>


Subject(s)
Animals , Male , Mice , Apolipoproteins E , Genetics , Blood Glucose , Metabolism , Diabetes Mellitus, Experimental , Blood , Drug Therapy , Diabetic Angiopathies , Drug Therapy , Drugs, Chinese Herbal , Pharmacology , Lipids , Blood , Mice, Knockout , RNA, Messenger , Genetics , Random Allocation , Thiazolidinediones , Pharmacology , Vascular Cell Adhesion Molecule-1 , Genetics , Metabolism
20.
West China Journal of Stomatology ; (6): 263-266, 2014.
Article in Chinese | WPRIM | ID: wpr-231871

ABSTRACT

<p><b>OBJECTIVE</b>This study aimed to observe the postoperative pain rate and degree of pain in preschool children with cleft lip and palate, and investigate the effect of nursing intervention on pain relief.</p><p><b>METHODS</b>A total of 120 hospitalized cases of three- to seven-year-old preschool children with cleft lip and palate were selected from May to October 2011. The subjects were randomly divided into the control group and experimental groups 1, 2, and 3. The control group used conventional nursing methods, experimental group 1 used analgesic drug treatment, experimental group 2 used psychological nursing interventions, and experimental group 3 used both psychological nursing intervention and analgesic drug treatment. After 6, 12, 24, and 48 h, pain self-assessment, pain parent-assessment, and pain nurse-assessment were calculated for the four groups using the pain assessment forms, and their ratings were compared.</p><p><b>RESULTS</b>The postoperative pain rates of the four groups ranged from 50.0% to 73.3%. The difference among the four groups was statistically significant (P < 0.001). The differences among the control group and experimental groups 1 and 2 were not statistically significant (P = 0.871), whereas the differences among experimental group 3 and the other groups were statistically significant (P < 0.001).</p><p><b>CONCLUSION</b>Postoperative pain in preschool children with cleft lip and palate is common. Psychological nursing intervention with analgesic treatment is effective in relieving postoperative pain.</p>


Subject(s)
Child, Preschool , Humans , Cleft Lip , General Surgery , Cleft Palate , General Surgery , Pain, Postoperative
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