2.Ginsenoside Rb1 affects the proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro**★
Zhijun LUO ; Hongmian LI ; Hegeng WANG ; Dalie LIU ; Hua NAN
Chinese Journal of Tissue Engineering Research 2013;(32):5799-5805
BACKGROUND:Various factors can affect the osteogenic differentiation of human adipose-derived stem cel s, and the osteoinductive factor of traditional Chinese medicine is very important for the research of human adipose-derived stem cel s. OBJECTIVE:To investagate the effects of ginsenoside Rb1 on the proliferation and osteogenic differentiation of human adipose-derived stem cel s in vitro. METHODS:The human adipose-derived stem cel s were isolated and cultured in vitro. After passaqed to the third generation, human adipose-derived stem cel s at 2×103/wel were incubated in a 96-wel plate, and treated with 200μL of 0.5, 1.0, 2.0, 4.0,6.0μmol/L ginsenoside Rb1 medium. The human adipose-derived stem cel s in the control group were treated with an equal volume of Dulbecco’s modified Eagle medium. Growth curves were examined by 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide T colorimetric assay. Alkaline phosphatase activity and osteocalcin content were detected by alkaline phosphatase kit and radio-immunity method, respectively. Calcified nodules were observed using alizarin red O staining. RESULTS AND CONCLUSION:The proliferation viability of human adipose-derived stem cel s was significantly increased after cultured with 0.5μmol/L ginsenoside Rb1. With the increasing of the concentration of ginsenoside Rb1, the mitogenic activity of the cel s was decreased. The 6.0μmol/L ginsenoside Rb1 showed a depressant effect on proliferation. Ginsenoside Rb1 could promote alkaline phosphatase activity and osteocalcin expression in human adipose-derived stem cel s and showed a dose-dependent manner. Calcified nodule formation induced by 4.6 and 6.0μmol/L ginsenoside Rb1 were better when compared with 0.5, 1.0 and 2.0μmol/L ginsenoside Rb1. Ginsenoside Rb1 can promote the proliferation of human adipose-derived stem cel s cultured in vitro in a certain concentration, and in the high concentration, the ginsenoside Rb1 can promote the osteogenic differentiation of human adipose-derived stem cel s. So ginsenoside Rb1 can be used as an osteoinductive factor.
3.Glutamine and tumor metabolism and their applications in nuclear medicine
Fei LIU ; Hua ZHU ; Xinfeng LIN ; Zheng LUO ; Nan LI ; Zhi YANG
Journal of International Oncology 2015;42(2):122-125
Many tumors depend on glutamine for energy.Glutamine metabolism is recognized as a distinctive feature in addition to Warburg effect in tumors.Nuclear medicine molecular tracer techniques represented by positron emission tomography (PET) provide a good means for early diagnosis and prognosis of tumors.PET imaging can detect glutamine metabolism region in tumors noninvasively,which can provide new diagnosis and therapeutics for glutamine-addicted tumors.
4.Promotive effect of recombinant human BIGH3 protein eye drops on the corneal epithelial healing in rabbit
Xin, LUO ; Hong-yan, GE ; Da-xi, XUE ; Nan, XIAO ; Dong-hua, QI ; Pei, TIAN ; Ping, LIU
Chinese Journal of Experimental Ophthalmology 2013;32(11):1006-1010
Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25% or 0.5% recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25% and 0.5% recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25%,0.5% recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5%recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25% recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25% and 0.5%recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25% and 0.5% recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.
5.The effect of intensive trunk muscle training on balance and walking in hemiplegic patients
Liang-Hua LIAO ; Xing-Mei JIANG ; Lin-Po LUO ; Zhi-Wei YE ; Bu-Zhi HUANG ; Nan-Yan XU ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(08):-
Objective To study the effect of intensive trunk muscle training on balance and walking in pa- tients with hemiplegia caused by stroke.Methods A total of 90 stroke patients were recruited and randomly divid- ed into a treatment group(45 cases)and a control group(45 cases).All the patients were given conventional reha- bilitation training.Meanwhile,intensive trunk muscle training was also administered for those in the treatment group as well.The trunk control function,balance ability and walking ability were assessed by using the trunk control test, Berg Balance Scale and the balance subscale of the Fugl-Meyer physical performance test,and Holden's functional ambulation classification,respectively,before and after 6 weeks of training.Results It was found that all the pa- tients scored better with the trunk control test,Berg's balance scale,the balance subscale of the Fugl-Meyer physical performance test and Holden's ambulation classification after treatment,and there were significant differences between the two groups after treatment(P
6.Association of CXCR4 and SDF-1alpha with organ-specific metastasis of nasopharyngeal carcinoma.
Hua-nan LUO ; Xiang-ping LI ; Xiong LIU ; Guo-hua ZHANG ; You-li LIU
Chinese Journal of Oncology 2009;31(4):260-264
OBJECTIVETo explore the role of CXCR4/SDF-1alpha axis in organ-specific metastasis of nasopharyngeal carcinoma (NPC) by assessment of CXCR4 expression in NPC cells and SDF-1alpha expression in distant target organs of NPC.
METHODSThirty patients with NPC and fifteen normal subjects were recruited in this study. The expressions of CXCR4 in NPC and normal cases were identified by RT-PCR and immunohistochemistry (IHC), then the relationship between CXCR4 expression and clinicopathological factors was analyzed. IHC was also used to analyze the SDF-1alpha protein expression in normal cervical lymph nodes (including normal superior and inferior deep cervical lymph nodes), bone marrow, lung, liver, kidney and colon tissues of NPC patients (5 cases/each group).
RESULTSThe relative expression level of CXCR4 mRNA in NPC (0.71 +/- 0.22) was significantly higher than that of normal nasopharynx tissues (0.14 +/- 0.07, F = 27.94, P < 0.05). The relative expression level of CXCR4 protein in NPC (1.58 +/- 0.59) was significantly higher than that of normal nasopharynx tissues (0.51 +/- 0.22, F = 17.75, P < 0.05). The high expression levels of CXCR4 mRNA and protein in NPC were closely related to clinical stage, cervical lymph node metastasis and cancer cell differentiation (P < 0.05). SDF-1alpha protein was strongly expressed in normal superior deep cervical lymph nodes, bone marrow, lung and liver (2.35 +/- 0.67), but absent or very poor expression in inferior deep cervical lymph nodes, kidney and colon tissues (0.68 +/- 0.23), and the differences between them were statistically significant (t = 10.13, P < 0.01).
CONCLUSIONCXCR4 is closely correlated to metastasis of nasopharyngeal carcinoma. CXCR4/SDF-1alpha axis may play an important role in organ-specific metastasis of NPC.
Adult ; Aged ; Bone Marrow ; metabolism ; Cell Differentiation ; Chemokine CXCL12 ; genetics ; metabolism ; Female ; Humans ; Liver ; metabolism ; Lung ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Receptors, CXCR4 ; genetics ; metabolism ; Young Adult
7.Alternative donor HSCT for 109 children with acquired severe aplastic anemia: a single center retrospective analysis.
Cheng Juan LUO ; Jing CHEN ; Jian Min WANG ; Xia QIN ; Bing Hua ZHANG ; Hua ZHU ; Xi Nan WANG ; Jiao Yang CAI ; Chang Ying LUO
Chinese Journal of Hematology 2020;41(2):128-131
Objective: To investigate the efficacy of alternative donor (AD) in the treatment of aplastic anemia (AA) in children. Methods: The clinical data of AA children who received AD HSCT in our center from Apr. 2010 to Dec. 2016 were retrospectively analyzed. The overall survival (OS) rate, implant success rate, incidence of acute and chronic graft-versus-host disease (GVHD) were statistically analyzed. Results: A total of 109 children with acquired AA, including 64 severe AA (SAA) , 32 very severe AA (VSAA) and 13 transfusion dependent non-severe AA (NSAA) , were recruited in this retrospective AD HSCT study, the median age was 6 (0.8-18) years old. Of them, 44 patients with 10/10 matched unrelated donor (MUD) , 44 patients with mismatched unrelated donor (MMUD) and 21 patients with mismatched related donor (MMRD) . All patients did not receive ATG before HSCT and the active infection was excluded. Except 3 patients suffered from a second graft failure (2 of them rescued by second HSCT) , 106/109 (97.2%) were engrafted with neutrophil and platelet recovery occurring at a median of 13 days (range, 9-19) and 16 days (range, 10-81) post-transplant. Until day 100 post transplantation, the incidence was 74.3% (81/109) for acute GVHD (aGVHD) and 39.4% (43/109) for grade Ⅱ-Ⅳ aGVHD, 30.7% (31/101) and 9.9% (10/101) for overall chronic GVHD (cGVHD) and moderate cGVHD, respectively, and nobody developed an extend cGVHD. After median follow up of 39 (0.7-103) months for all patients, 13 of 109 patients died. The estimated 5-year overall survival (OS) of the entire cohort was 88.1% (95%CI 81.1%-91.4%) with no difference among the MUD, MMUD and MMRD cohort (93.2%, 84.1% and 85.7%, respectively, P=0.361) . Conclusion: These excellent outcomes suggest that unmanipulated AD PBSC is a good HSCT source for children with SAA. It's reasonable to consider AD HSCT as first line therapy for SAA children without matched sibling donor. Better strategies are required to prevent GVHD.
Adolescent
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Anemia, Aplastic
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Child
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Child, Preschool
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Graft vs Host Disease
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Hematopoietic Stem Cell Transplantation
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Humans
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Infant
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Retrospective Studies
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Tissue Donors
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Treatment Outcome
8.MiR-20 Regulates Myocardiac Ischemia by Targeting KATP Subunit Kir6.1
NIE LI ; ZHAO YA-NAN ; LUO HONG-YAN ; HU XIN-WU ; ZHANG LIANG-PIN ; LIANG HUA-MIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):486-490
This study aimed to examine the functional role of microRNA-20 (miR-20) and its potential target,Kir6.1,in ischemic myocardiocytes.The expression of miR-20 was detected by real-time PCR.Myocardiocytes were stained with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) reagent for apoptosis evaluation.Western blotting was used to detect the Kit6.1 protein in ischemic myocardiocytes transfected with miR-20 mimics or inhibitors.Luciferase reporter gene assay was performed to confirm the targeting effect of miR-20 on KCNJ8.The results showed that miR-20 was remarkably down-regulated,while the KATP subunit Kir6.1 was significantly up-regulated,during myocardial ischemia.The miR-20 overexpression promoted the apoptosis of ischemic myocardiocytes,but showed no such effect on normal cells.Under ischemic condition,myocardiocytes transfected with miR-20 mimics expressed less Kir6.1.On the contrary,inhibiting miR-20 increased the expression of Kir6.1 in the cells.Co-transfection of miR-20 mimics with the KCNJ8 3’-UTR plasmid into HEK293 cells consistently produced less luciferase activity than transfection of the plasmid alone.It was concluded that miR-20 may regulate myocardiac ischemia by targeting KATP subunit Kir6.1 to accelerate the cell apoptosis.Therefore miR-20 may serve as a therapeutic target for myocardial ischemic disease.
9.Detection of new mutations in the dystrophin gene by denaturing high-performance liquid chromatography.
Ya-nan CHEN ; Xin ZHOU ; Chun-lian JIN ; Yan XU ; Chang-kun LIN ; Li-hua CAO ; Ning LI ; Xue ZHANG ; Yang LUO
Chinese Journal of Pediatrics 2007;45(6):413-416
OBJECTIVEDuchenne muscular dystrophy (DMD) is an X-linked recessive disease caused by dystrophin gene mutations; 55%-65% of these pathogenic mutations are large deletion and duplication mutations that can be detected by multiplexed polymerase chain reaction. However, finding the remaining micro-mutations (substitutions, deletions or insertions of one or several nucleotides) cannot be achieved in this way. The aim of the present study was to detect mutations of the dystrophin gene in individuals with Duchenne muscular dystrophy (DMD) by denaturing high-performance liquid chromatography (DHPLC) and to establish a rapid and sensitive screening platform for micro-mutations leading to DMD.
METHODSTwenty patients negative for large deletions in the dystrophin gene by multiplex PCR were selected for further screening by DHPLC and 20 normal male without DMD family history as the control cohort. Dystrophin exons and their flanking sequences were individually amplified by genomic PCR and the amplicons showing abnormal DHPLC profile were directly sequenced to identify the position and the type of the mutations.
RESULTSAfter screening 68 exons covering the two deletion hotspots and 3'UTR region, four pathogenic mutations, including c.6808_6811del TTAA, c.4959_4960insA, c.8656C > T and c.8608C > T, were found in four DMD patients. Moreover, c.6808_6811del TTAA, c.4959_4960ins and c.8656C > T have not been reported previously. The first two frameshift mutations were predicted to produce premature stop codons, p.Leu2270MetfsX9 and p.Ser1654LysfsX5, respectively. The remaining two were nonsense mutations, leading to p.R2886X and p.R2870X, respectively.
CONCLUSIONThree novel and one recurrent dystrophin mutations have been identified in Chinese DMD patients. This study has demonstrated that DHPLC is an effective screening method for micro-mutation associated with DMD.
Chromatography, High Pressure Liquid ; methods ; trends ; DNA Mutational Analysis ; Dystrophin ; genetics ; Humans ; Infant ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Sequence Deletion
10.Effects of low level manganese exposure on the serum neuroendocrine hormones in the welders.
Chan WANG ; Ji-pei LU ; Yue-ming JIANG ; Nan-hua MA ; Wei-ping QIN ; Hai-lan LUO ; Yan-ni HUANG ; Hai-bin CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(2):94-97
OBJECTIVETo study the effects of low level manganese (Mn) exposure on the serum neuroendocrine hormones levels of the welders.
METHODSThe exposure group consisted of 41 male welders, 40 male workers without exposing to harmful agents served as controls. The serum contents of prolactin (PRL), luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (TST) and thyroid stimulating hormone (TSH) of 81 subjects were detected by chemiluminescence immunoassay.
RESULTSThe geometric mean value of airborne Mn concentrations was 0.03 mg/m(3) (0.003 - 0.519 mg/m(3)) in the welding circumstances. The levels of Mn in red blood cells (RBCs) and urinary Mn of the exposure group were significantly higher than those of control group (P < 0.01). The contents of serum LH and TSH of the exposure group were 2.89 ± 0.69 mIU/ml and 1.45 ± 0.56 uIU/ml, which were significantly lower than those (3.82 ± 1.61 mIU/ml and 2.19 ± 1.28 µIU/ml) of control group (P < 0.01). The serum contents of LH, FSH and TSH of the group exposed to Mn for < 5 years were significantly lower than those of the control group, The serum TST level of the group exposed to Mn for < 5 years was significantly higher than those of the control group and group exposed to Mn for 5 ∼ years, the serum FSH level of the group exposed to Mn for < 5 years was significantly lower than that of the group exposed to Mn for 10 years (P < 0.05 or P < 0.01). The serum contents of LH and TSH of the group exposed to Mn for 5 ∼ years were significantly lower than those of the control group (P < 0.05 or P < 0.01). The serum contents of PRL, LH and TSH of the group exposed to Mn for 10 years were significantly lower than those of the control group (P < 0.05). There was negative correlation between blood (RBC) Mn and urinary Mn (r = -0.310, P < 0.05), also there was negative correlation between serum PRL and serum TST (r = -0.409, P < 0.01), the positive correlation between serum LH and serum FSH was observed (r = 0.361, P < 0.05).
CONCLUSIONThe results of present study showed that the long exposure to low level of Mn may decrease the levels of serum PRL, LH and TSH in workers occupationally exposed to Mn, which can influence the metabolism of neuroendocrine hormones to certain extent.
Adult ; Air Pollutants, Occupational ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Manganese ; Occupational Exposure ; Prolactin ; blood ; Testosterone ; blood ; Thyrotropin ; blood ; Welding