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Objective:To explore the action mechanism of suppressing expression of mitogen- activated protein kinase 14（MAPK14）to alleviate glutamate excitatory toxicity and its neuronal protection effect.Methods:Lentivirus-mediated MAPK14 interference vector was synthetized by Shanghai Jikai Gene Chemical Technology Co.，Ltd. Astrocytes were obtained from SD rats 48 hours after birth，which were cultured in vitro and transfected by lentivirus-mediated transfection. According to the random number table，the cells were divided into three groups：（1）un-transfected group（normal group）with normal astrocytes and the cells were cultured in regular medium composed of Dulbecco's?modified Eagle's?medium（DMEM）；（2）negative control group with astrocytes transfected by MAPK14 no-loaded interference vector；（3）lentivirus transfected group with astrocytes transfected by MAPK14 interference vector. Seventy-two hours after transfection，astrocytes were co-cultured with neurons for 48 hours，and then they were cultured in a medium containing glutamate for 2 hours. The detection indexes included the optimal multiplicity of infection（MOI）value for astrocytes transfected by lentivirus vector，mRNA levels of MAPK14 and glial glutamate transporter 1（GLT-1）detected by rPCR 72 hours after transfection，protein levels of MAPK14 and GLT-1 detected by Western blot 72 hours after transfection，level of lactate dehydrogenase（LDH）and mortality of neurons measured by spectrophotometry and flow cytometry 2 hours after culturing in the medium with glutamate. Results:（1）The optimal MOI value for lentivirus transfecting astrocytes was 30，and astrocytes grew well after transfection.（2）Seventy-two after transfection，the mRNA level of MAPK14 in lentivirus transfected group（0.005 7±0.000 6）was significantly decreased as compared with un-transfected group（0.013 1±0.001 1）and negative control group（0.013 9±0.001 0）（ P<0.01），the mRNA level of GLT-1 in lentivirus transfected group（0.009 1±0.001 2）was not significantly changed as compared with un-transfected group（0.008 7±0.000 3）and negative control group（0.008 9±0.001 1）（ P>0.05）.（3）Seventy-two hours after transfection，the protein level of MAPK14 in lentivirus transfected group（0.29±0.04）was significantly decreased as compared with non-transfected group（0.61±0.05）and negative control group（0.63±0.01）（ P<0.01），the protein level of GLT-1 in lentivirus transfected group（0.73±0.06）was significantly increased as compared with un-transfected group（0.20±0.03）and negative control group（0.23±0.09）（ P<0.01）.（4）After astrocytes were co-cultured with neurons and subsequently cultured in the medium containing glutamate for 2 hours，the level of LDH in lentivirus transfected group［（109.67±2.40）U/L］was significantly lower than that in un-transfected group［（141.52±3.88）U/L］and negative control group［（141.29±3.61）U/L］（ P<0.01）. The mortality of neurons in lentivirus transfected group［（38.72±0.26）%］was significantly lower than that in un-transfected group［（52.94±1.36）%］and negative control group［（54.30±1.23）%］（ P<0.01）. Conclusions:The transfection with lentivirus-mediated MAPK14 interference vector can increase expression of GLT-1 in astrocytes to increase glutamate re-uptake and relieve the glutamate excitatory toxicity in neurons，which may provide a new experimental basis for future use of astrocyte gene regulation to alleviate neuronal injury caused by glutamate excitatory toxicity after traumatic brain injury.
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Objective@#To compare the clinical and imaging characteristics of traumatic acute subdural hematoma acute subdural hematoma with rapid resolution and those without rapid resolution.@*Methods@#A retrospective case-control analysis was conducted on the clinical data of 60 traumatic acute subdural hematoma patients with hematoma thickness≥5 mm admitted to Second Affiliated Hospital of Shantou University Medical College from January 2011 to May 2018. There were 37 males and 23 females, aged 18-80 years [(47.0±16.9)years]. There were 27 patients in the rapid resolution group and 33 patients in the non-rapid resolution group. Coagulation function [prothrombin time (PT) and international normalized ratio (INR)] on admission, hospital stay, Glasgow outcome scale (GOS), and brain CT results were compared between the two groups.@*Results@#The PT and INR values in the rapid resolution group were (11.9±2.1)s and 1.1±0.2 respectively, while those in the non-rapid resolution group were (10.8±1.0)s and 1.0±0.1 respectively, with significant differences (P<0.05). There was no significant difference in hospital stay and GOS between the two groups (P>0.05). The thickness of subdural hematoma of the two groups in the first CT scanning was (8.2±2.3)mm and (7.3±1.8)mm, respectively, with no statistically significant difference (P>0.05). In the second CT scanning, the hematoma thickness in the rapid resolution group was significantly lower than that in the non-rapid resolution group [(2.7±1.9)mm vs. (6.6±2.1)mm] (P<0.01). The incidence of low density zone between the hematoma and intracranial plate was statistically higher in rapid resolution group than that in non-rapid resolution group (93% vs. 36%) (P<0.01). The incidence of subarachnoid hemorrhage(SAH) increase was significantly higher in rapid resolution group than that in non-rapid resolution group in the second CT scan (74% vs. 15%) (P<0.01). The conversion rate of acute subdural hematoma to subacute or chronic subdural hematoma was 4% in the rapid resolution group, which was significantly lower than 18% in the non rapid resolution group (P<0.05).@*Conclusions@#The abnormal coagulation function and low density zone indicated by CT are two important indicators of rapid resolution in patients with traumatic acute subdural hematoma. The risk of conversion from acute into subacute or chronic subdural hematoma is lower in rapid resolution of traumatic acute subdural hematoma, suggesting that rapid resolution may be one of the prognostic indicators of ASDH patients.
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Objective To compare the clinical and imaging characteristics of traumatic acute subdural hematoma acute subdural hematoma with rapid resolution and those without rapid resolution.Methods A retrospective case-control analysis was conducted on the clinical data of 60 traumatic acute subdural hematoma patients with hematoma thickness ≥ 5 mm admitted to Second Affiliated Hospital of Shantou University Medical College from January 2011 to May 2018.There were 37 males and 23 females,aged 18-80 years [(47.0±16.9)years].There were 27 patients in the rapid resolution group and 33 patients in the non-rapid resolution group.Coagulation function [prothrombin time(PT)and international normalized ratio(INR)] on admission,hospital stay,Glasgow outcome scale(GOS),and brain CT results were compared between the two groups.Results The PT and INR values in the rapid resolution group were(11.9±2.1)s and 1.1±0.2 respectively,while those in the non-rapid resolution group were(10.8±1.0)s and 1.0±0.1 respectively,with significant differences(P＜0.05).There was no significant difference in hospital stay and GOS between the two groups(P＞0.05).The thickness of subdural hematoma of the two groups in the first CT scanning was(8.2±2.3)mm and(7.3±1.8)mm,respectively,with no statistically significant difference(P＞0.05).In the second CT scanning,the hematoma thickness in the rapid resolution group was significantly lower than that in the non-rapid resolution group [(2.7±1.9)mm vs.(6.6±2.1)mm](P＜0.01).The incidence of low density zone between the hematoma and intracranial plate was statistically higher in rapid resolution group than that in non-rapid resolution group(93％vs.36％)(P＜0.01).The incidence of subarachnoid hemorrhage(SAH)increase wassignificantly higher in rapid resolution group than that in non-rapid resolution group in the second CT scan(74％vs.15％)(P＜0.01).The conversion rate of acute subdural hematoma to subacute or chronic subdural hematoma was 4％in the rapid resolution group,which was significantly lower than 18％in the non rapid resolution group(P＜0.05).Conclusions The abnormal coagulation function and low density zone indicated by CT are two important indicators of rapid resolution in patients with traumatic acute subdural hematoma.The risk of conversion from acute into subacute or chronic subdural hematoma is lower in rapid resolution of traumatic acute subdural hematoma,suggesting that rapid resolution may be one of the prognostic indicators of ASDH patients.
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Objective To improve clinically the recognition of fungal infection associated with hemophagocytic lymphohistiocytosis (HLH) in children. Methods Clinical data of 3 children with fungal infection complicated with HLH in our hospital was retrospectively analyzed. Results All the 3 cases complained of recurrent fever, 2 cases with cough and one case with vomiting. Hepatosplenomagaly and lymphadenectasis were found in the medical examination. The time of diagnosis of fungal infection through etiological examination was 5 days after admission. It was further diagnosed as hemophagocytic lymphohistiocytosis after failure of effective antifungal therapy. Routine blood test showed the counts of leukocytes were increased in early stage, while the number of platelets and hemoglobin decreased in different degrees. The recovery is not satisfactory using antifungal therapy alone, and 2 of them are gradually aggravated and treated with mechanical ventilation. On the basis of antifungal therapy, 2 cases were treated under HLH-2004 regimen, 1 received dexamethasone treatment. All the 3 cases received intravenous immune globulin, and showed improvement. Conclusions Fungal infection complicated with HLH in childhood is rare. The effect of simple antifungal therapy on the progression is limited. However, increasing immunosuppressive therapy based on effective antifungal therapy can improve the prognosis.
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Objective Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening condition characterized by excessive inflammation, with a high incidence in children and a death rate of 40%.This study was to analyze the gene expression profile in child-hood HLH and explore the important pathways of childhood HLH using bioinformatic methods . Methods The childhood HLH gene ex-pression profile data GSE26050 were obtained from the Gene Expression Omnibus (GEO) database of the National Center for Biotechnolo-gy Information.Differentially expressed genes were identified with the GEO 2R online analysis tools released recently .The key pathways of the differentially expressed genes were investigated using the Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway enrichment a-nalysis. Results A total of 184 differentially expressed genes were identified , 126 upregulated and the other 58 downregulated .They were enriched in 3 pathways, including cytokine-cytokine receptor interaction , hematopoietic cell lineage and NOD-like receptor signaling pathways. Conclusion Bioinformatic tools allow the identification of the key genes and pathways associated with the development and progression of childhood HLH and point out the potential directions for researches on the mechanisms of childhood HLH .
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Objective To investigate the effect of allgeneic hematopoietic stem cell transplantation (allo-HSCT) for β-thalassemia major. Methods Twenty-four β-thalassemia major patients with median age of 4 years (range: 2～15 years), 18 boys and 6 girls, received allo-HSCT.They were classified into class Ⅱ-Ⅲ according to Pesaro thalassemia classification. Twenty-three transplantations were from sibling donor and 1 was from mother, either HLA-identical (n = 23) or HLA-mismatched (5/6) (n = 1). Fifteen patients received bone marrow transplantation (BMT) plus peripheral blood stem cell transplantation (PBSCT), and 9 were subjected to umbilical cord blood transplantation (UCBT). The conditioning regimen consisted of busalphan, cyclophosphamide,fludarabine, plus hydroxyurea before transplantation. Graft-versus-host disease (GVHD) prophylaxis included CsA, methotrexate, antilymphpcute globulin, and mycophenolate mofetil. The median follow-up period was 13 months (range: 3～69). Results Of 24 patients, there were 21 cases (87. 5 %) of disease-free survival, 1 (4. 2 %) transplantation-related death, and 2 cases (8. 3 %) of rejection. Three-year overall survival and disease-free survival rate was 91.7 % and 87. 5 %respectively. The cumulative incidence of grade Ⅱ -Ⅳ acute GVHD and chronic GVHD was 16. 7 %and 20. 3 %, particularly cumulative extensive chronic GVHD was 5. 0 %. Conclusion The sibling donor BMT plus PBSCT is an effective and safe way to treat β-thalassemia major. Cord blood is an important source of hematopoietic stem cells for HSCT. The protocol GVHD prophylaxis of CsA,MTX, ATG with a low-dose and short course of MMF can effectively reduce the incidence of severe acute GVHD, improve the outcome of thalassemia transplantation.
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Objective To evaluate X-ray and MRI features of limbs in childhood acute leukemia.Methods Thirteen children with acute leukemia in our pediatric hematology ward were recruited.Allpatients were pathologically diagnosed by bone marrow aspiration and complained of bone or joint pain in the first visit.ConventionaI X-ray and MRI examinations of algesic sites were performed before clinical treatment and after complete remission.MR images were obtained with SE-T1WI,SE-T2WI and T2WI-fat suppressed sequences and symmetria bilateralis was requested while scanning.X-ray and MRI manifestations were evaluated and compared.Resuits All 13 patients had received X-ray examinations.Among them,6 had normal X-ray findings,whereas the other 7(14 sites)showed various abnormalities including radiolucent metaphyseal bands(5 sites),periosteal reaction(3 sites),osteapenia(2 sites),mixed lesions(lysissclerosis,1 site),and permeative pattern(3 sites).The number of patients for MRI examinations was 8(11 sites).Among them,6(9 sites)showed bone marrow infiluration and bone marrow necrosis accompanied by normal X-ray findings,another 2(2 sites)showed bone marrow infiltration associated with radiographic abnormalities of periosteal reaction and radiolucent metaphyseal bands.Four cases were followed up within 1 week when reached complete remission by chemotherapy.MR images features included reduced sizes of bone marrow infiltration lesions associated with increased signal intensity on T1WI,and disappearance of double-line sign on bone marrow necrosis accompanied by signal homogenization.However,the radiograph before and after treatment in the same cases did not differ significantly.Conclusions MRI was earlier and more comprehensive in showing limbs bone marrow abnormality than radiogram in acute leukemia children with chief complaint of osteoarticular pains.MRI might be one of indicators in following up therapeutic effect for AL children with osteoarticular disorder.
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Objective: To investigate the efficacy of specific point acupuncture and manipulative adjustment on muscle strength of the knee osteoarthritis. Method: One hundred and five patients were randomly allocated to two groups. Sixty-eight cases in the treatment group were treated by specific point acupuncture and manipulative adjustment on muscle strength of the knee joint. While 37 cases in the control group were treated with specific point acupuncture alone. Result: The total effective rate was significantly higher in the treatment group than in the control group (P＜0.05). Conclusion: Specific point acupuncture and manipulative adjustment on muscle strength of the knee joint has a satisfactory curative effect on knee osteoarthritis.
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@#ObjectiveTo evaluate the assistant effct of electroacupuncture on acute incomplete spinal cord injury treated with routine therapy.Methods9 cases who accepted routine therapy, including operation and medicine, were in control group, while other 7 cases who accepted electroacupuncture other than routine therapy were in treating group.ResultsThere were 2 cases were effective, 4 cases were improved and 3 cases were not improved in control group, while 6 cases were clinical recovery, 1 case was effective in treating group.ConclusionElectroacupuncture can improve the effect of routine therapy on acute incomplete spinal cord injury.
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@# 目的观察神经外科与针刺联合治疗急性脊髓不完全损伤的疗效。方法神经外科治疗组根据指征确定是否进行手术,药物治疗均给予激素、脱水、抗去甲肾上腺素类药物、神经营养药物,高压氧。神经外科与针刺联合治疗组除上述神经外科治疗外,采用针刺治疗,针刺处方:电针刺激夹脊穴,根据不同辨证配合远端取穴。结果神经外科治疗组9例,持续治疗 3—6个月,出院后随访6个月,6例好转,肌力有所改善,最好者可达到肌力IV—IV+级,3例无效,甚至退步,瘫痪进一步加重。神经外科与针刺联合治疗组病人5例,经针刺及电针治疗10—80天时间后,4例临床治愈,1例经针刺治疗好转后,因转入骨科手术治疗腰椎压缩性骨折时脱离针刺治疗。结论神经外科与针刺联合治疗急性脊髓损伤效果好,并能抑制脊髓损伤的进一步恶化。
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Objective To investigate the effects of acupuncture on gene expression profile of neurotrophin and its receptors in cerebral cortex of neonatal rats after cerebral hypoxic-isehemic injury,and to explore the molecu- lar mechanism of acupuncture in treatment of neonatal hypoxic-ischemic encephalopathy.Methods The model of cerebral hypoxic-ischemic injury was established with 10 neonatal Sprague-Dawley(SD)rats,who were subjected to acupuncture once daily for 14 days.The animals were sacrificed on the next day of the last acupuncture and their brain cortex was sampled for examination of gene expression,using GEArray Q series neurotrophin and receptors gene array.Results After 14 days of acupuncture,it was found that 48 genes(50% of total genes on the microarray) were expressed differently between the two groups,of which 40 genes(83.3% of differently expressed genes),such as those of the brain-derived neurotrophic factor(BDNF),ciliary neurotrophic factor(CNTF),ciliary neurotrophic factor receptor(CNTFR),basic fibroblast growth factor(bFGF)and fibroblast growth factor receptor type 1 (FGFR1)were up-regulated,and 8 genes(16.7% of differently expressed genes),such as genes of neuregulin-1 (Nrg1),neuregulin-4(Nrg4)and tyrosine kinase C(TrkC)were down-regulated.Conclusion Acupuncture can regulate the expression of various genes of neurotrophin and receptors in cerebral cortex of neonatal rats after cerebral hypoxic-ischemic injury,which might be the mechanism of acupuncture facilitating the recovery of the rats from hy- poxic-ischemic encephalopathy.
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Purpose To study the reversal effect on multidrug resistance (MDR) by TNF-α gene combined with verapamil (VRP) or tamoxifen (TAM). Methods By using recombinant retrovirus vector, TNF-α gene was transfected into multidrug-resistant human breast cancer cell line MCF7/ADR. The TNF-α secreting cell clone MCF7/ADR-TNF was obtained by G418 screening. The integrating and secreting of TNF-α were analyzed by PCR and ELISA. MTT assay and formula"I = d/D1 + d/D2" were used to evaluate the reversal effect of multidmg resistance with TNF-α gene combined with verapamil or tamoxifen. ResultsThe level of TNF-α secreted by MCF7/ADR-TNF was 1 737 pg/ml (106cells/48 h). Compared with control,the resistance to ADR of MCF7/ADR-TNF was reversed by 1.6 times. The reversal effect produced by combination of TNF-α gene and VRP was antagonistic. The combination of TNF-α gene and TAM produced synergic effect (interaction index I = 0.64). ConclusionsTNF-α gene combined with TAM has synergic effect on reversing MDR.
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Paraffin - embedded tissue of skin biopsy specimens taken retrospectively from 24 patients with cutaneous malignant lymphomas(CML) and 8 patients with cutaneous benign lymphoid infiltrates (CLI) and other dermatoses were estimated with PC10 immunostaining. Our results show a statistical significant difference between PC10 indices for cutaneous genuine histiocytic lymphoma(CGHL), cutaneous germinal center cell-derived lymphomas(CGCCL), cutaneous peripheral T-cell lymphomas(CPTL) other than mycosis fungoides(MF) and Sezary's syndrome (SS),and MF when compared with those for CLI. There is a linear relationship between PC10 index and square root of PC10 density, both of which seem to have a parallel relationship to the seventy of malignancy in CML. The nuclear volume of the positive tumor cell or lymphocyte with PC10 immunostaining may be also useful in differentiating CML from CLI.