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1.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 1443-1449, 2021.
Article in Chinese | WPRIM | ID: wpr-923814

ABSTRACT

Objective To systematically review the effects of exercise on systemic inflammation of chronic obstructive pulmonary disease (COPD) and skeletal muscle dysfunction. Methods The literature about the effect of exercise on COPD systemic inflammation and skeletal muscle dysfunction were retrieved from PubMed, Web of Science, CNKI, VIP and Wanfang data, until June, 2021, supplemented by reference review and manual retrieval. Results A total of 192 literatures were retrieved and eight were included, involving 245 subjects. The comprehensive results showed that exercise could decrease the level of pro-inflammatory factors and increase the level of anti-inflammatory factors. Exercise can improve the motor ability and skeletal muscle structure of patients with COPD. Exercise can improve systemic inflammation of COPD, which is related to the mode, intensity and duration of exercise. Exercise may affect ubiquitin-protease, insulin-like growth factors -1/phosphatidylinositol 3 kinase/Akt and other pathways by regulating the inflammatory response, and improve skeletal muscle dysfunction. Conclusion Exercise has certain effect on reducing systemic inflammation and improving skeletal muscle dysfunction.

2.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 1443-1449, 2021.
Article in Chinese | WPRIM | ID: wpr-923798

ABSTRACT

Objective To systematically review the effects of exercise on systemic inflammation of chronic obstructive pulmonary disease (COPD) and skeletal muscle dysfunction. Methods The literature about the effect of exercise on COPD systemic inflammation and skeletal muscle dysfunction were retrieved from PubMed, Web of Science, CNKI, VIP and Wanfang data, until June, 2021, supplemented by reference review and manual retrieval. Results A total of 192 literatures were retrieved and eight were included, involving 245 subjects. The comprehensive results showed that exercise could decrease the level of pro-inflammatory factors and increase the level of anti-inflammatory factors. Exercise can improve the motor ability and skeletal muscle structure of patients with COPD. Exercise can improve systemic inflammation of COPD, which is related to the mode, intensity and duration of exercise. Exercise may affect ubiquitin-protease, insulin-like growth factors -1/phosphatidylinositol 3 kinase/Akt and other pathways by regulating the inflammatory response, and improve skeletal muscle dysfunction. Conclusion Exercise has certain effect on reducing systemic inflammation and improving skeletal muscle dysfunction.

3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 32-41, 2021.
Article in Chinese | WPRIM | ID: wpr-906016

ABSTRACT

Objective:To explore the effects of Huatan Tongluo Decoction (HTTLD) on the morphology and function of brain tissues and intestine in rats with cerebral ischemia/reperfusion based on the gut-brain axis. Method:Sixty SPF male rats were randomly divided into a sham operation group, a model group, high- (28.66 g·kg<sup>-1</sup>), medium- (14.33 g·kg<sup>-1</sup>), and low-dose (7.16 g·kg<sup>-1</sup>) HTTLD groups, and an edaravone (4 g·kg<sup>-1</sup>)+<italic>Clostridium butyricum</italic> (5.0×10<sup>8</sup> cfu·mL<sup>-1</sup>) group. The model was established by focal cerebral ischemia/reperfusion in rats. The drugs were administered by gavage. The brain tissue injury was determined by neurological deficit score and 2,3,5-triphenyl tetrazolium chloride (TTC) staining. The effect of cerebral ischemia/reperfusion on intestinal motility was assessed by the propulsion rate of small intestine. The intestinal mucosal cell damage was evaluated by the pathomorphological examination of the duodenal mucosa. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of <italic>D</italic>-lactate (<italic>D</italic>-LAC), diamine oxidase (DAO), and bacterial endotoxin (lipopolysaccharide, LPS) in serum. Western blot was used to detect the expression of Occludin, Claudin-5, and zonula occludens 1 (ZO-1) in the duodenum. Result:After cerebral ischemia/reperfusion, rats developed neurological deficit symptoms. The neurological deficit score in the model group was higher than that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the high- and medium-dose HTTLD groups could relieve the symptoms of neurological deficits and lower neurological deficit scores (<italic>P<</italic>0.01). The results of TTC staining showed that the model group presented obvious infarcts in brain tissues compared with the sham operation group (<italic>P<</italic>0.01). The cerebral infarction volumes of HTTLD groups were reduced compared with that in the model group (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. Furthermore, the propulsion rate of small intestine in the model group was significantly reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, HTTLD groups could increase propulsion rates of small intestine (<italic>P<</italic>0.01), especially the high-dose HTTLD group, and the effect was dose-dependent. After cerebral ischemia/reperfusion, obvious duodenal mucosal damage could be observed, which was relieved after the administration of HTTLD. Western blot results showed that the protein expression of ZO-1, Occludin, and Claudin-5 in the model group was reduced compared with that in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups could up-regulate the expression of ZO-1, Occludin, and Claudin-5 to varying degrees (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. ELISA showed that the serum <italic>D</italic>-LAC, DAO, and LPS of the model group were elevated compared with those in the sham operation group (<italic>P<</italic>0.01). Compared with the model group, the HTTLD groups showed reduced <italic>D</italic>-LAC and DAO (<italic>P<</italic>0.05, <italic>P<</italic>0.01), and the medium- and high-dose HTTLD groups showed reduced LPS (<italic>P<</italic>0.05, <italic>P<</italic>0.01), especially the high-dose HTTLD group. Conclusion:After cerebral ischemia/reperfusion, the rats showed damaged brain tissues, neurological dysfunction, intestinal mucosal injury, weakened intestinal motility, and destroyed the intestinal mucosal barrier. HTTLD can protect against brain-gut axis injury after cerebral ischemia/reperfusion by reducing the damage on brain tissues and gastrointestinal mucosa, relieving the symptoms of neurological deficits, promoting gastrointestinal motility, improving intestinal barrier function, and reducing the release of intestinal bacterial metabolites or poisons.

4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 73-79, 2021.
Article in Chinese | WPRIM | ID: wpr-905959

ABSTRACT

Objective:To investigate the effect of astragaloside Ⅳ(AST Ⅳ)and Notoginseng total saponins (NTS) combined with bone marrow mesenchymal stem cell (BMSC) transplantation on neural repair and angiogenesis in rats with cerebral ischemia. Method:The rats were randomly divided into a sham operation group, a model group, low- and high-dose AST Ⅳ + NTS groups, a BMSC infusion group, and low- and high-dose BMSC infusion+AST Ⅳ (10 and 20 mg·kg<sup>-1</sup>) + NTS group (25, 50 mg·kg<sup>-1</sup>). BMSCs were isolated and purified by whole bone marrow adherent culture. The positive expression of surface markers of BMSCs (CD29, CD90, CD34, and CD45) was detected by flow cytometry. The focal cerebral ischemia model was established by middle cerebral artery occlusion (MCAO). The PKH26-labeled BMSCs were injected into the tail vein of rats in the BMSC infusion group, once a day. The rats in the combination groups received BMSC injection once a day and intragastric administration of drugs twice a day. Other groups were administered twice a day by gavage. The sham operation group and the model group received the same amount of normal saline. Symptoms and signs of neurological deficits were assessed by the Longa method and the cerebral infarction rate was determined by TTC staining. The survival and vascularization [double positive expression of PKH26/vascular endothelial growth factor (VEGF)] after transplantation of BMSCs were observed by the immunofluorescence method. The protein expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was measured by Western blot. Result:BMSCs were properly isolated and cultured. The identification of surface markers CD29, CD90, CD34, and CD45 was consistent with the characteristics of BMSCs. The neurological deficit score and cerebral infarction rate of the model group were significantly increased (<italic>P</italic><0.01). All drugs and cell transplantation could alleviate the above pathological changes in varying degrees. The strongest effect was observed in high-dose BMSC infusion+AST Ⅳ+NTS group (<italic>P</italic><0.01), which was superior to those in the AST Ⅳ+NTS groups or the BMSC infusion group. BMSC injection helped cells survive in the ischemic brain tissues and promoted angiogenesis, and this effect could be enhanced by the combination with drugs. After cerebral ischemia, the expression of Ang1 and TGF-<italic>β</italic><sub>1</sub> was increased, and the effect in the BMSC infusion+AST Ⅳ+NTS groups was the strongest (<italic>P</italic><0.01). Conclusion:AST Ⅳ combined with NTS can promote the survival of transplanted BMSCs and facilitate angiogenesis after target repair of damaged blood vessels after cerebral ischemia. The mechanism may be related to the improvement of the local microenvironment in the brain after cerebral ischemia and the promotion of the survival and differentiation of transplanted stem cells.

5.
Shanghai Journal of Preventive Medicine ; (12): 793-2020.
Article in Chinese | WPRIM | ID: wpr-862455

ABSTRACT

Objective To evaluate the implementation process, and cost-effectiveness of fluoride varnish in preventing primary caries project for children aged 3-6 years in Huangpu District.This would provide the basis for the popularization of fluoride varnish project in preschool children as a basic measure of caries prevention in community health service. Methods From 2016 to 2017, children aged 3-6 years old from 12 kindergartens in Huangpu District were varnished by fluoride twice a year.Three-years-old children in 12 kindergartens were divided into intervention group and control group according to the baseline survey results.The intervention group was implemented according to the established norms, while the control group was implemented according to the routine requirements.Follow-up examinations were conducted after 4 interventions to monitor primary caries increment among these children. Results Caries rate, average caries index and SiC in children aged 3, 4, 5 and 6 years were all dropped after fluorination in year of 2018.The caries rate of 5-year-old deciduous teeth decreased from 58.8% in 2015 to 45.0% in 2018.The dental caries rate in the intervention group was lower than that in the control group, and the frequency of fluoride application per capita was higher than that in the control group, showing a statistically significant difference (P < 0.05), but there was no significant difference in the rate of fluoride application and average caries index between the two groups (P > 0.05).The direct input-output ratio of the project is 1 : 5.16. Conclusion Fluorinated caries prevention project using fluoride varnish for preschool children can effectively reduce the incidence of dental caries in deciduous teeth and has good economic benefits.The application of standard fluorine coating on preschool children′s deciduous teeth can affect caries prevention.

6.
Chinese Traditional and Herbal Drugs ; (24): 1649-1656, 2019.
Article in Chinese | WPRIM | ID: wpr-851236

ABSTRACT

Objective To investigate whether borneol can promote the bioactive components of the combination of astragaloside IV (AST IV) and Panax notoginseng saponins (PNS) into the blood-brain barrier of rats with middle cerebral artery occlusion (MCAO)/reperfusion. Methods Using the model of MCAO/reperfusion, rats were randomly divided into sham-operation group, model group, borneol group, AST IV group, PNS group, AST IV + PNS group and borneol + AST IV + PNS group, and the content of AST IV and the bioactive components of PNS (ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1) in the cerebral cortex and the cerebellum of the affected side and the healthy side were determined by liquid chromatography-mass spectrometry (LC-MS/MS). Results AST IV, whether used alone or combined with PNS and borneol, was mainly distributed in the cerebral cortex after oral administration, especially in the affected cerebral cortex. Borneol combined with AST IV and PNS significantly increased the content of AST IV in the affected and the healthy cerebral cortex. The bioactive components of PNS such as ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 was mainly distributed in the affected side of the cerebellum when PNS was used alone. Borneol combined with AST IV + PNS significantly increased the content of ginsenoside Rb1 in the cerebral cortex, especially in the affected cortex, increased the content of Rg1 in the healthy and the affected cortex, and increased the content of notoginsenoside R1 in the cerebral cortex, especially in the affected cortex, as well as in the cerebellum. Conclusion AST IV and the bioactive components of PNS such as ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 have a certain distribution in the cerebral cortex and the cerebellum after cerebral ischemia-reperfusion in rats. AST IV was mainly distributed in the cerebral cortex when it was used alone, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 were mainly distributed in the cerebellum when PNS was used alone. The combination of borneol combined with AST IV and PNS can promote the gather of AST IV, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 to the cerebral cortex, especially to the cortex of the ischemia-reperfusion side; Moreover, it can promote the absorption of AST IV, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 in the cerebral cortex to varying degrees, especially in the affected cortex.

7.
Chinese Traditional and Herbal Drugs ; (24): 5795-5801, 2019.
Article in Chinese | WPRIM | ID: wpr-850674

ABSTRACT

Objective: To observe the effects of astragaloside IV (AST IV) combined with Panax Notoginseng saponins (PNS) on proliferation, apoptosis, migration and neuronal differentiation of oxygen glucosedeprivation/reoxygenation model rat bone marrow mesenchymal stem cells (BMSCs). Methods: BMSCs were isolated, cultured, amplified and purified by the whole bone marrow adherent method. The positive expression rates of BMSCs surface markers, CD29, CD90, CD34, and CD45 were detected by flow cytometry. The third generation of BMSCs was pretreated with AST IV and PNS doses of high (100 μmol/L + 60 μmol/L), medium (50 μmol/L + 30 μmol/L), and low (25 μmol/L + 15 μmol/L) for 24 h. The model of ischemia-reperfusion injury was established by OGD/R. Meanwhile, the normal group (BMSCs were cultured normally) and the model group (OGD/R was used to establish an ischemia reperfusion injury model) were established. The cell increment rate was detected by CCK-8 assay. Cell apoptosis was detected by flow cytometry. Transwell assay was used to detect the migration of BMSCs. The condition of BMSCs differentiation into neurons and astrocytes was observed by Nestin/NSE and Nestin/GFAP immunofluorescence double labeling. Results: BMSCs were successfully cultured and separated, and the positive rates of CD29 and CD90 detected by flow cytometry were 94.23% and 94.69%, while the positive rates of CD34 and CD45 were 5.76% and 5.31%. Compared with the normal group, the survival rate of the model group was reduced significantly and the apoptosis rate was increased significantly (P < 0.05). Compared with the model group, the combination of different doses of AST IV and PNS could promote the proliferation of BMSCs (P < 0.05, 0.01) and inhibit the apoptosis (P < 0.05, 0.01). Compared with the normal group, the model group and the AST IV and PNS group at different doses could promote the migration of BMSCs (P < 0.05). Compared with the model group, the number of migrated cells in the AST IV and PNS groups at different doses was increased significantly (P < 0.05). Compared with the normal group, the model group and the AST IV and PNS groups at different doses could promote the differentiation of BMSCs into neurons and astrocytes (P < 0.01). Compared with the model group, the positive expression rates of Nestin/NSE and Nestin/GFAP in the AST IV and PNS groups at different doses were increased significantly (P < 0.01). Conclusion: AST IV combined with PNS can promote the proliferation and migration of BMSCs of ischemia-reperfusion model in vitro, inhibit the apoptosis, and induce their directional differentiation into neurons and astrocytes.

8.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 1070-1074, 2019.
Article in Chinese | WPRIM | ID: wpr-905124

ABSTRACT

Objective:To explore the clinical effect of group occupational training based on cognitive behavior on patients with post-stroke cognitive impairment. Methods:From May, 2018 to March, 2019, 56 patients with post-stroke cognitive impairment were randomly divided into control group (n = 29) and experimental group (n = 27). Both groups received routine rehabilitation, and the experimental group received group occupational therapy, three times a week, for twelve times. They were assessed with Montreal Cognition Assessment (MoCA), modified Barthel Index (MBI) and the Short Form of Health Survey (SF-36) before and after intervention. Results:The scores of MoCA, MBI and SF-36 increased in both groups after intervention (t > 2.275, P < 0.05), and their difference before and after intervention was more in the experimental group than in the control group (t > 2.835, P < 0.01). Conclusion:Group occupational training can improve the cognitive function of patients with post-stroke cognitive impairment and improve their activities of daily living and quality of life.

9.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 410-415, 2018.
Article in Chinese | WPRIM | ID: wpr-702507

ABSTRACT

Neuromuscular electrical stimulation(NMES)is one of the electrotherapy that exerts a low-frequency current over the targeted nerves and muscles to induce skeletal muscle contractions to treat skeletal muscle dysfunction in patients.As an alternative rehabilitation modality,NMES is essential for patients with chronic obstructive pulmonary disease(COPD) who are unable to participate in conventional rehabilitation program.NMES could effectively improve the skeletal mus-cle strength,endurance,exercise capacity and quality of life of the patients with COPD.It may be related to remodeling skeletal muscle structure,reducing oxidative stress in skeletal muscle,and improving balance of skeletal muscle protein metabolism.

10.
Chinese Journal of Current Advances in General Surgery ; (4): 757-760, 2017.
Article in Chinese | WPRIM | ID: wpr-703761

ABSTRACT

Objective:To purpose of this study is to introduce how peripheral blood neutrophil/lymphocyte ratio (NLR) before operations influences the prognosis of patients with breast cancer.Methods:Review of systems were used to analyze patients who suffered from breast cancer and accepted modified radical mastectomy of breast cancer according to the clinical data of 180 cases of Shenyang Military Region General Hospital between January 2002 and January 2005.All the patients were classified into two groups according to the NLR with the critical value at 6.0.2 statistics were used to evaluate the relationship between NLR of two groups and clinical pathological characteristic.Kaplan-Meier survival analysis and Cox's proportional hazards regression model were used to analyze the relationship among NLR of two groups,other clinical pathologic characteristic and prognosis of patients.Results:The high level of NLR is related with the size of patients' tumor,lymph node metastasis and TNM stages (P<0.05).Kaplan-Meier survival curves indicated the group of high level of NLR's overall survival (OS) and disease-free survival (DFS) was significantly lower than the low level NLR group (P<0.05).Single factor and multivariate cox's proportional hazards regression model indicated the high level of NLR before operations,the size of tumor,lymph node metastasis and TNM stages were significantly related with the OS and DFS (P<0.05).Conclusion:The high level of NLR before operations is an independent risk factor to influence the OS and DFS of the patients who accepted modified radical mastectomy of breast cancer.

11.
Journal of Experimental Hematology ; (6): 896-903, 2017.
Article in Chinese | WPRIM | ID: wpr-271898

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of human umbilical cord blood-derived mesenchymal stem cells(HUCMSC) on the leukemic cell line HL-60 and acute lymphoblastic leukemia cell line Jurkat as well as the role of CXCL12/CXCR4.</p><p><b>METHODS</b>HL-60 cells and Jurkat cells were co-cultured with human umbilical cord blood mesenchymal stem cell (HUCMSC), and the model was treated with G-CSF, AMD3100 and their combination. The cell viability and cell cycle were measured by Cell Counting Kit-8 (CCK-8), the apoptosis and the cell-cycle analysis were assessed by flow cytometry with the Annexin V/PI double staining. The expression of surface CXCR4 protein and total CXCR4 protein of leukemic cells were detected by flow cytometry and Western blot respectively.</p><p><b>RESULTS</b>HUCMSC could decrease the viability of HL-60 cells and Jurkat cells, as well as the percentage of apoptotic cells, they could also increase the number of G/Gcells, while G-CSF and AMD3100 could reduce the proliferation of HL-60 cells and Jurkat cells in HUCMSC co-culture model, destructed the anti-apoptotic effect of HUCMSC on HL-60 cells and Jurkat cells, and the combination of 2 drugs resulted in a synergistic effect. The G-CSF could reduce the expression of surface CXCR4 protein and total CXCR4 protein in leukemic cells, while AMD3100 could only decrease the expression of surface CXCR4 protein of leukemia cell membrane, having no effect on the expression of CXCR4 protein in cytoplasm.</p><p><b>CONCLUSION</b>Human umbilical cord blood mesenchymal stem cells can inhibit the proliferation and apoptosis of acute leukemia cells and increase the number of G/Gphase cells in leukemic cells. The AMD3100 can decrease the expression of surface CXCR4 protein in leukemia cells, G-CSF can decrease expression of total CXCR4 protein as well as membrane CXCR4 protein. Both of them can block the CXCL12/CXCR4 signal axis, weakening the relationship between leukemia cells and microenvironment. And on the basic of HUCMSC influenced leukemia cells' growth and proliferation, the cell viability will be weakened, its apoptosis will be promoted, and the percentage of G/Gphase cells in leukemia cells will be decreased.</p>

12.
China Journal of Chinese Materia Medica ; (24): 3786-3794, 2017.
Article in Chinese | WPRIM | ID: wpr-335783

ABSTRACT

The aim is to study the effect of astragaloside Ⅳ (AST Ⅳ) combined with Panax notoginseng saponins (PNS) on cerebral ischemia-reperfusion injury, and to probe the synergistic mechanism through the pharmacokinetics of the four major components such as AST Ⅳ, ginsenoside Rg₁ (Rg₁), ginsenoside Rb₁ (Rb₁), notoginsenoside R₁ (R₁) in cerebral ischemia-reperfusion rats. Following the establishment of cerebral ischemia/reperfusion model in rats by modified suture method, neurological function score, cerebral infarction area and pathomorphology were used to evaluate the pharmacological effect that the combination of AST Ⅳ and PNS antagonized cerebral ischemia-reperfusion injury; the contents of AST Ⅳ, Rg₁, Rb₁, R₁ in rat plasma of different time points were determined with ultra performance liquid chromatography tandem massspectrometry (UPLC-MS/MS), pharmacokinetic parameters were calculated and pharmacokinetics changes of the main effective components were analyzed. The results showed that AST Ⅳ, PNS alone and their combination could reduce the cerebral infarction area of rats, relieve the behavioral scores of neurologic deficit, improve the pathological changes after cerebral ischemia, the effects of the combination were better. Among AST Ⅳ, Rg₁, Rb₁, R₁, the area under the curve (AUC) was significantly increased, the mean residence time of (MRT0-t) was delayed, the peak concentration (Cmax) was significantly raised, the apparent volume of distribution (Vz/F) was reduced, and the clearance rate in vivo was significantly slowed. It suggested that AST Ⅳ combined with PNS has synergistic enhancement on anti-cerebral ischemia/reperfusion injury, moreover, make the pharmacokinetic behavior of the main effective components change, the mechanism may be associated with prolonging the retention time of the effective components in cerebral ischemia condition, elevating the bioavailability.

13.
Journal of Experimental Hematology ; (6): 326-331, 2016.
Article in Chinese | WPRIM | ID: wpr-360091

ABSTRACT

<p><b>OBJECTIVE</b>To investigate HOXB4, PRDM16 and HOXA9 gene expression in patients with acute myeloid leukemia (AML) and its clinical significance.</p><p><b>METHODS</b>Real-time quantitative PCR (RT-qPCR) with SYBR Green assay was used to detect the expression of HOXB4, PRDM16 and HOXA9 gene in AML patients (40 cases), the patients with complete remission (9 cases) and patients with non-malignant hematologic diseases as control (10 cases). The relationship between the expression levels of gene HOXB4, PRDM16, HOXA9 and clinical features was investigated by statistical analysis.</p><p><b>RESULTS</b>The gene expression levels of HOXB4, PRDM16, HOXA9 in newly diagnosed or relapsed AML patients were significantly higher than those in patients with non-malignant hematologic disease (P < 0.05). It was observed that the expression of HOXB4 gene in newly diagnosed or relapsed patients positively correlates with leukemic blasts in bone marrow (r = 0.39). The expression levels of HOXB4, PRDM16 and HOXA9 positively correlate with each other. There was statistical significance among gene expressions in different phases (newly diagnosed, relapse, remission). No correlation was observed between expression levels of HOXB4, PRDM16, HOXA9 and chromosome risk status. It was noticed that expression levels of HOXB4, PRDM16, HOXA9 genes were lower in the patients achieved remission after two courses of chemotherapy than those in the other. And high expression group of each gene had a lower remission rate than that in the low expression group.</p><p><b>CONCLUSION</b>The expression level of HOXB4, PRDM16, HOXA9 genes and leukemic blasts somewhat correlate with curative effect and prognosis. The expression of HOXB4, PRDM16, HOXA9 genes is higher in newly diagnosed and relapsed leukemia patients, and lower in the patients acquired CR/PR. High expression of HOXB4, PRDM16, HOXA9 genes predicts an adverse prognosis.</p>


Subject(s)
Humans , Bone Marrow , Case-Control Studies , DNA-Binding Proteins , Genetics , Metabolism , Gene Expression , Homeodomain Proteins , Genetics , Metabolism , Leukemia, Myeloid, Acute , Genetics , Metabolism , Prognosis , RNA, Messenger , Metabolism , Real-Time Polymerase Chain Reaction , Recurrence , Remission Induction , Transcription Factors , Genetics , Metabolism
14.
Journal of Experimental Hematology ; (6): 1578-1584, 2013.
Article in Chinese | WPRIM | ID: wpr-264972

ABSTRACT

This study was purposed to investigate the difference of nucleated cell (NC) count, CD34(+) cell ratio and expansion multiple, cell cycle and colony formation capability in in vitro expanded human umbilical cord blood CD34(+) cells from HOXB4-transfecting directly and HOXB4-transfected human umbilical cord mesenchymal stem cells (HUCMSC) by means of prepared feeder layers of HUCMSC. The HUCMSC were divided into 2 groups:first group, in which HOXB4 gene was transfected into HUCMSC by using lentiviral vecfor, and feeder layers were set up; and second group in which feeder layers for HUCMSC of non-transfected HOXB4 gene were set up. The CD34(+) cells were separated from HUCB by magmatic activated cell sorting(MACS). After culture in medium with cytokines for 2 days, CD34(+) cells were divided into 5 groups, including control group and experimental group. The control groups included CD34(+) cells as group A (blank control group) and GFP-CD34(+) cells as group B (negative control group) and experimental groups included HOXB4-CD34(+) cells as group C, HUCMSC+CD34(+) cells as group D, HOXB4-HUCMSC+ CD34(+) cells as group E and cells in all groups were cultured in vitro. The number of nucleated cells were counted at day 6, 10, 14 of culture and CD34 immunophenotypes, cell cycle and colony forming capability were measured at day 10 of culture in different conditions. The results indicated that HOXB4 gene could be transfected into HUCMSC by lentiviral vector and feeder layers were set up successfully. After culture for 14 days, the nucleated cells in 5 groups could be amplified effectively, and the expansion levels in 5 groups were in order HOXB4-HUCMSC+CD34(+) cell group> HOXB4-CD34(+) cell group>HUCMSC+CD34(+) cell group> control groups (P < 0.05). At day 10 of in vitro expansion the CD34(+) cell percentage decreased significantly in all groups, while the number of CD34(+) cell increased in experiment groups, which were in order HOXB4-CD34(+) cells group> HOXB4-HUCMSC+CD34(+) cell group>HUCMSC+CD34(+) cell group>control groups (P < 0.05). The cell cycle detection showed that the percentage of cells in S+G2/M phase in experiment groups were higher than that in control groups (P < 0.05), and percentage of cells in HOXB4-HUCMSC+CD34(+) cells group was higher (41.57%) than that in HOXB4-CD34(+) cells group(37.87%) and HUCMSC+CD34(+) cell group (28.65%) (P < 0.05). There was no statistical difference in the CFU number between HOXB4-HUCMSC+CD34(+) cell group and HOXB4-CD34(+) cell group, which were both higher than that in HUCMSC+CD34(+) cell group and control groups (P < 0.05).It is concluded that the CD34(+) cells cultured on HOXB4-HUCMSC feeder layers can be amplified significantly and kept the characteristics of stem cells, The feeder lager of HOXB4-HUCMSC is relative safe for amplification of CD34(+) cells in vitro, it possesses the potential useful value.


Subject(s)
Humans , Antigens, CD34 , Allergy and Immunology , Cell Separation , Cells, Cultured , Fetal Blood , Cell Biology , Homeodomain Proteins , Genetics , Mesenchymal Stem Cells , Cell Biology , Allergy and Immunology , Transcription Factors , Genetics , Transfection , Umbilical Cord , Cell Biology , Allergy and Immunology
15.
Chinese Journal of Hematology ; (12): 1010-1014, 2013.
Article in Chinese | WPRIM | ID: wpr-295751

ABSTRACT

<p><b>OBJECTIVE</b>To investigate miR- 125b regulation mechanism by identifying miR-125b target genes and its function in acute myeloid leukemia (AML).</p><p><b>METHODS</b>The bioinformatics software and database were applied to predict and analyze target genes of miR-125b. The vector contained the target gene 3'-UTR portion cloned into a luciferase reporter construct. A luciferase reporter assay was performed following co-transfection of small molecular miR-125b mimics and target gene wild-type or mutant plasmid into HEK-293T cells. Further in leukemia cell lines NB4 and HL-60, the protein level of target gene was measured by Western blot after overexpression miR-125b. Finally, the viabilities of NB4 and HL-60 cells were measured by CCK-8 assay at 24 h, 48 h, 72 h, 96 h after electroporation.</p><p><b>RESULTS</b>Bcl-2-antagonist/killer 1 (Bak1), a pro-apoptotic gene, was a target gene of miR-125b by software predicts. Reporter vector containing the 3'-UTR Bak1 wild and mutation sites were co-transfected with small molecule analogues of miR-125b in HEK-293T cells. Dual luciferase reporter gene assay system showed that miR-125b significantly suppresses the reporter gene activity containing Bak1 3'-UTR by about 53.8% (P<0.05), but it didn't suppresses the reporter gene activity containing 3'-UTR Bak1 mutation. Western blot showed that miR-125b mimics significantly down-regulated the expression of Bak1 in human leukemia cell lines NB4 and HL-60. Meanwhile, the growth rate of cells treated with miR-125b obviously increased compared with that in control by CCK-8 test (P<0.05).</p><p><b>CONCLUSION</b>Our findings strongly indicated that BAK1 was a downstream target gene of miR-125b, and miR-125b promoted proliferation in human AML cells at least partially by targeting Bak1, so we speculated that miR-125b as an oncogene could be a potential therapeutic target for treating AML.</p>


Subject(s)
Humans , Cell Line, Tumor , Cell Proliferation , Genetic Vectors , HEK293 Cells , HL-60 Cells , MicroRNAs , Genetics , Transfection , bcl-2 Homologous Antagonist-Killer Protein , Genetics , Metabolism
16.
National Journal of Andrology ; (12): 235-238, 2012.
Article in Chinese | WPRIM | ID: wpr-238991

ABSTRACT

<p><b>OBJECTIVE</b>To establish a method for internal quality control (IQC) of sperm concentration test in the laboratory.</p><p><b>METHODS</b>We set the concentrations of frozen semen at 20 x 10(6) and 80 x 10(6) as low and high concentrations of putative IQC products, with QC-BEADSTM quality control beads (QCBs) as the control. Using the double-blind method, four technicians determined the sperm concentrations of the IQC products and QCBs by computer-assisted sperm analysis, and drew a quality control chart (Xbar chart and Sbar chart) for each product. Through a month of continuous detection, we calculated and compared the intra- and inter-batch coefficients of variation (CV%) of the quality control products of high and low concentrations.</p><p><b>RESULTS</b>The intra-batch coefficients of variation of the assumed IQC products of high and low concentrations were CV3.5% and CV2.4%, and their inter-batch coefficients of variation were CV10.2% and CV9.6%. The intra-batch coefficients of variation of the QCBs of high and low concentrations were CV5.1% and CV7.1%, and their inter-batch coefficients of variation were CV7.1% and CV8%. The intra-batch coefficients of variation of both IQC products and QCBs of high and low concentrations were <10%, and their inter-batch coefficients of variation were <15%, which conformed to Levey-Jennings quality control principles and achieved IQC purposes. No significant differences were found in either intra- or inter-batch coefficients of variation between the IQC products and QCBs of high and low concentrations (P>0.05), indicating that assumed IQC products can replace QCBs for internal quality control in the laboratory.</p><p><b>CONCLUSION</b>The IQC method we established for determining sperm concentration is simple, feasible and reliable.</p>


Subject(s)
Humans , Male , Double-Blind Method , Quality Control , Semen Analysis , Methods , Reference Standards , Semen Preservation , Sperm Count , Sperm Motility , Spermatozoa
17.
Chinese Journal of Hematology ; (12): 98-102, 2012.
Article in Chinese | WPRIM | ID: wpr-345932

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of bone marrow-derived mesenchymal stem cells (MSC) from a third party donor for secondary poor graft function (PGF) following allogeneic hematopoietic stem cell transplantation(allo-HSCT).</p><p><b>METHODS</b>Five patients with secondary PGF were treated with MSC at a dose of 1 x 10(6)/kg body weight at a median of 47 days (35 to 61) after secondary PGF. MSC were derived from bone marrow (BM) of HLA-disparate third party donors, cultured in vitro and infused without HSC. If absolute neutrophil cell (ANC) and platelet counts (PLT) did not reach the standardization of > 1.5 x 10(9)/L and > 50.0 x 10(9)/L, respectively, within 28-30 days after the first MSC treatment, a second MSC treatment was required.</p><p><b>RESULTS</b>MSC were infused once in one patient and twice in four patients with an interval of 28 to 30 days. All patients obtained ANC and PLT recovery at a median of 34 (25 to 49) days and 47 (26 to 54) days, respectively, without toxic side effects within follow-up periods of median 761 (204-1491) days. Three patients developed Epstein-Barr virus (EBV) reactivation at 42, 48, 108 days after MSC infusion, respectively and two of the three coverted to posttransplant lymphoproliferative disorders (PTLD).</p><p><b>CONCLUSION</b>MSC from a third party donor are effective to patients with secondary PGF following allo-HSCT, whether it might increase the risk of EBV reactivation and EBV-associated PTLD need further observation.</p>


Subject(s)
Adolescent , Female , Humans , Male , Young Adult , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Mesenchymal Stem Cell Transplantation , Methods , Transplantation, Homologous
18.
Chinese Journal of Hematology ; (12): 178-181, 2011.
Article in Chinese | WPRIM | ID: wpr-252001

ABSTRACT

<p><b>OBJECTIVE</b>To screen childhood ALL related microRNAs (miRNAs), analyze association of miRNAs expression profiles with prognosis and relapse in childhood acute lymphoblastic leukemia (ALL) and explore new indicator for predicting relapse and prognosis.</p><p><b>METHODS</b>miRNAs expression profile was analyzed by gene chip in 49 newly diagnosed childhood ALL and 12 primary immune thrombocytopenia (ITP) cases (as control group). Abnormal expression of miRNAs was verified by qRT-PCR. The correlation of miRNAs expression pattern with indicators predicting early prednisone response and relapse within a year was analyzed.</p><p><b>RESULTS</b>Specific expression of miRNAs profiles associated with prednisone response and early relapse in childhood ALL was identified. Eight miRNAs (miR-18a, miR-532, miR-218, miR-625, miR-193a, miR-638, miR-550 and miR-633) could distinguish prednisone sensitive from insensitive. The early relapse of newly diagnosed patients with either high-risk or non-high-risk clinical types had some characteristics of abnormal expression of miRNAs, including miR-7, miR-216 and let-7i upregulated, while miR-486, miR-191, miR-150, miR-487 and miR-342 downregulated.</p><p><b>CONCLUSIONS</b>The initial screening reveals miRNAs differentially expressed from normal in ALL suggesting the potential roles of them in leukemogenesis. MiRNAs expression signatures may be useful for predicting prognosis and relapse in childhood ALL and directing personalized treatment.</p>


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , MicroRNAs , Genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Diagnosis , Genetics , Pathology , Prognosis , Recurrence , Transcriptome
19.
Acta Physiologica Sinica ; (6): 529-534, 2010.
Article in Chinese | WPRIM | ID: wpr-337716

ABSTRACT

In this study, we investigated the mechanism of linoleic acid-stimulated increase in intracellular calcium concentration ([Ca(2+)](i)) in pancreatic islet β-cells. Pancreatic islet cells were primarily isolated from rats and cultured for the experiments. The cells were loaded with Fluo-3/AM, the indicator of [Ca(2+)](i), and the intensity of Fluo-3 was measured using confocal microscope. The islet β-cells were identified by immunocytochemical staining with insulin antibody after recording. The drugs were given by perfusion system. The results showed that linoleic acid (20 μmol/L) stimulated [Ca(2+)](i) increase with the first peak increase and the following plateau increase. Linoleic acid-stimulated [Ca(2+)](i) increase was partly inhibited by removal of extracellular calcium and by transient receptor potential (TRP) channel blocker, La(3+), and it was totally blocked by exhaustion of intracellular calcium stores and inhibition of phospholipase C. It is concluded that linoleic acid stimulates [Ca(2+)](i) increase in islet β-cells through both extracellular calcium influx via TRP channels and calcium release from intracellular calcium stores.


Subject(s)
Animals , Male , Rats , Calcium , Metabolism , Insulin-Secreting Cells , Cell Biology , Metabolism , Linoleic Acid , Pharmacology , Primary Cell Culture , Rats, Sprague-Dawley , Transient Receptor Potential Channels
20.
Journal of Experimental Hematology ; (6): 1019-1022, 2007.
Article in Chinese | WPRIM | ID: wpr-318798

ABSTRACT

This study was aimed to investigate whether mesenchymal stem cells (MSCs) existed in human umbilical cord vein and to establish the methods of isolation and expansion in vitro. The MSCs derived for perfusion of umbilical cord vein (UVMSCs) were collected after parturition of Healthy pregnant women. The morphology of MSCs and their differentiation potential into osteoblast, adipocyte and chondrocyte were observed the phenotype and cell cycle of MSCs were determined by using flow cytometry. The result showed that the mesenchymal stem cells separated from umbilical cord vein gave rise to a population of adherent cells with a typical fibroblast-like morphology. Similarly to bone marrow-derived MSCs, they highly expressed CD29, HLA-ABC, CD166, CD105, CD73 and CD44, and were negative for any hematopoietic and endothelial markers (CD45, CD34, CD14 and CD144). Functionally, they could differentiate into the osteoblast, adipocyte and chondrocyte. It is concluded that MSCs exist in the human umbilical cord vein perfusion. Their read amplification in vitro contribute to clinical applications for cell therapy and tissue engineering.


Subject(s)
Female , Humans , Cell Differentiation , Cell Separation , Methods , Cells, Cultured , Fetal Blood , Cell Biology , Mesenchymal Stem Cells , Cell Biology , Umbilical Veins , Cell Biology
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