italic>γ-Aminobutyric acid (GABA) is a crucial inhibitory neurotransmitter found in various cells in the human body. While the GABAergic system is typically associated with the nervous system, recent research has revealed that immune cells and tumor cells also express components of this system. In the tumor microenvironment (TME), GABA is secreted to act extracellularly on other cells. GABA is metabolized via the GABA shunt and is involved in the tricarboxylic acid (TCA) cycle by generating succinate, which can provide energy for tumor cells. Activation of GABA receptors (GABARs) is a major pathway through which GABA participates in the regulation of antitumor immune responses. The activation of GABA type A receptors (GABA_ARs) can inhibit the activation and proliferation of T cells, elicit anti-inflammatory macrophages, and promote tumor cell growth and migration, while activation of GABA type B receptors (GABA_BRs) is generally considered to inhibit cancer cell migration and induce cancer cell apoptosis. In general, receptor activation inhibits immune cells, but the effect on tumor cells varies. Additionally, the downregulation of the expression levels of GABA transporters (GATs) is involved in tumor progression. Although antagonists of GABA metabolism and drugs that act on GABA receptors are considered therapeutic drugs for tumors, there have been few clinical studies conducted on them.

Objective To evaluate the impact of total parenteral nutrition(TPN)on nutrition status and inflammatory markers in hospitalized fasted patients with inflammatory bowel disease(IBD).Methods A retrospective study was performed and 82 hospitalized fasted IBD patients [male/female=58/24,(39.4±14.5)years] who received TPN entered the study.Among them,38 patients had ulcerative colitis(UC)and 44 patients suffered from Crohn`s disease(CD).Clinical data(gender,age,duration of disease,history of disease,prednisone,immuno-suppressor,and antibiotics)were obtained from medical records.Nutritional parameters,C-creative protein(CRP),and erythrocyte sedimentation rate(ESR)before and after TPN were also obtained.Average caloric supplementation by TPN was(4 437.3±1 199.1)kJ/d and the nitrogen amount was(9.9±1.7)g/d.Median PN length was 15 days(7-54 days).67 IBD patients received a TPN formula with glutamine(≥14 d,25 patients vs.0-14 d,42 patients)and 15 IBD subjects received TPN without glutamine.Malnutrition was diagnosed by body mass index(BMI)and serum albumin level.Results The prevalence of undernutrition was 90.2％(74/82)in the study population.CD patients had a significantly longer history of disease [84(3-288)months vs.24(1-324)months,P<0.001] and a significantly lower BMI [(15.6±1.8)kg/m2 vs.(19.1±3.5)kg/m2,P<0.001] compared with those in UC patients.TPN improved nutritional parameters [serum albumin:(28.7±6.6)g/L before TPN vs.(31.7±5.8)g/L after TPN,P<0.001;pre-albumin:(174.1±85.5)mg/L before TPN vs.(227.2±82.8)mg/L after TPN,P<0.001].Conclusions TPN improves nutritional status in hospitalized fasted IBD patients.However,prospective randomized controlled trials are required to estimate the role of low-to-middle dosage of glutamine in IBD patients.

BACKGROUNDInvasive pulmonary aspergillosis (IPA) is a severe and frequently fatal disease in patients receiving treatment with immunosuppressive agents such as cyclophosphamide. Aspergillus fumigatus (A. fumigatus) now is a leading cause of IPA. Dectin-1 and Toll-like receptor 2 (TLR2) are important pattern recognition receptors involved in immune responses to A. fumigatus in vitro. However, the expression of the two receptors during the infection of A. fumigatus in vivo is not completely understood. The effects of cyclophosphamide treatment on the expression of the receptors need to be further studied.

METHODSWe established different immune status in mice models with or without A. fumigatus infection. On days 1, 3 and 5 post inoculation, pulmonary tissues from mice of the different groups were harvested. Dectin-1 and TLR2 mRNA expression in the lungs of the mice were investigated by real-time PCR. The pulmonary fungal burden in the mice with A. fumigatus infection was also evaluated.

RESULTSIn the immunocompetent mice, three days after A. fumigatus inoculation, dectin-1 and TLR2 expression increased markedly compared with the normal control group. Cyclophosphamide inhibited the clearance of pathogens and the expression of dectin-1 with or without A. fumigatus infection in the lungs as well. There was no statistical difference in TLR2 expression between the different immune status groups.

CONCLUSIONSOur results suggest that in vivo, dectin-1 and TLR2 are activated during the experimental period which would provide a broad range of possibilities for a specific and effective inflammatory response to kill A. fumigatus. Inhibition of dectin-1 expression may be one of the mechanisms of cyclophosphamide in the development of IPA.

Animals ; Aspergillosis ; immunology ; microbiology ; Aspergillus fumigatus ; immunology ; physiology ; Cyclophosphamide ; pharmacology ; Immunosuppressive Agents ; pharmacology ; Lectins, C-Type ; Lung ; drug effects ; metabolism ; microbiology ; Male ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins ; genetics ; Polymerase Chain Reaction ; Toll-Like Receptor 2 ; genetics

OBJECTIVETo detect the expression of B7-H1 gene in bone marrow mononuclear cells (BMMNCs) from leukemia patients and explore its clinical implications.

METHODSThe B7-H1 mRNA expression levels of BMMNCs from 74 newly diagnosed leukemia patients and 10 normal volunteers were detected by real-time quantitative PCR. At the same time, BMMNCs from 12 patients in complete remission (CR) after chemotherapy and 5 in relapse were followed up. The correlation between the clinical features of 74 de novo leukemia patients and the expression level of B7-H1 gene was analyzed.

RESULTSThe mRNA expression level of B7-H1 gene in BMMNCs from de novo leukemia patients (RQ = 0.125) was lower than that from normal control (RQ=1). When patients achieved CR the gene expression level (RQ = 69.07) was significantly higher than that before CR (P = 0.001). After relapsed, its level (RQ=4) was still higher than that before CR (P > 0.05). No clinical parameters such as gender, age, peripheral white blood count, blast cells ratio in BM, CD34 positive cells were significantly correlated with the expression level of B7-H1 except the response to therapy. The initial expression level of B7-H1 gene in non CR patients after therapy was significantly higher than that in CR patients (RQ = 26. 91, P = 0.005).

CONCLUSIONThe mRNA expression level of B7-H1 gene in newly diagnosed leukemia patients is lower than that in normal controls, and is higher in CR patients than in newly diagnosed patients. There is a correlation between the gene expression level and responsiveness to therapy.

Adult ; Antigens, CD ; metabolism ; B7-H1 Antigen ; Female ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVESTo obtain high therapeutic effect and low toxicity single-chain immunotoxin against hepatocellular carcinoma (HCC).

METHODSHuman mutant tumor necrosis factor-alpha (mTNFalpha) was linked with the 3' end of humanized single-chain Fv against HCC (hscFv25) in pGEX4T-1 vector. The anti-HCC immunotoxin was expressed in Escherichia coli and identified by western blot. The primary tumor regression trial in nude mice bearing HCC was evaluated the targeting therapeutic value of hscFv25-mTNFalpha. The tumor tissues were stained by immunohistochemical with TNFalpha antibody.

RESULTSThe expression of single-chain immunotoxin hscFv25-mTNFalpha was 12% of total bacteria proteins. The tumor regression trials of hscFv25-mTNFalpha showed 5/5 effective. It had 2/5 completely remission and 3/5 partly remission. The therapeutic result of hscFv25-mTNFalpha was better than that of mTNFalpha (F=8.70, 0.05). The HCC tissue treated by hscFv25-mTNFalpha expressed TNFalpha positive reaction. The positive granule mainly existed in HCC cytoplasm.

CONCLUSIONThe single-chain immunotoxin hscFv25-mTNFalpha has high therapeutic effect and low toxicity. It has potentialities for clinical application.

Animals ; Immunoglobulin Fragments ; therapeutic use ; Immunohistochemistry ; Immunotoxins ; therapeutic use ; Liver Neoplasms, Experimental ; therapy ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; therapeutic use

BACKGROUND: Elastic intramedullary nail is commonly used in the treatment of fractures of children, but few studies concern the elastic intramedullary nail for treating fractures in adults. OBJECTIVE: To investigate the repair effect of elastic intramedullary nail in the treatment of 22-A fracture in forearm of adults. METHODS: From January 2015 to April 2016, a total of 21 adult patients with the 22-A fracture (35 fractures) were treated with manipulative reduction and elastic intramedullary nail fixation in the First Affiliated Hospital of Guangzhou University of Chinese Medicine. The follow-up time was 12-18 months. Radiographs were taken and the guidance of the limb function training was given at regular intervals. The Andserson scoring system was used to evaluate the patients' forearm limb function. Fracture healing, elbow, wrist joint activity and forearm rotation were recorded. The satisfactory questionnaires were recorded. The patients were divided into three grades as satisfaction, general satisfaction and dissatisfaction; simultaneously, reasons were recorded. RESULTS AND CONCLUSION: (1) The Andserson scoring was satisfactory in 16 cases accounted for 76%, general satisfaction in 3 cases accounted for 14%, dissatisfaction in 2 cases accounted for 10%. (2) Wrist joint activity increased from (172±4)° before the operation to (181±3)° at the end of the follow-up. Elbow joint activity increased from (102±18)° before the operation to (124±13)° at the end of the follow-up. Forearm rotation activity increased from (84±11)° before the operation to (155±13)° at the end of the follow-up (P < 0.05). (3) In the follow-up of the 21 patients, 13 patients were satisfied with the result of surgery; 5 patients were generally satisfied; 2 patients were dissatisfied because the limited limb functions; and 1 patient was dissatisfied because of the nail irritability; the dissatisfaction rate accounted for 14%. (4) Elastic intramedullary nail can obtain affirmative effect in the treatment of adult 22-A fracture of the forearm; and clinical application should be based on the type of fracture.

This study was aimed to investigate various factors influencing long-term survival in patients with acute promyelocytic leukemia. A single institutional retrospective study with long-term follow-up was performed to better define the prognostic factors and a rationale for the use of ATRA, chemotherapy, and As(2)O(3) in the treatment of newly diagnosed APL patients. Newly diagnosed patients with APL entering complete remission (CR) were followed up for 6 to 185 months (n = 170) from January 1990 to December 2004. Univariate and multivariate analysis of 8 potential factors influencing survival and prognosis were carried out with Log-Rank and Cox regression method, including sex, age, initial WBC count, the level of lactic hydrogenase (LDH), first induction regimen, time from induction therapy to CR, post-remission therapy, negative or positive rate of PML-RAR alpha and follow-up of reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the estimated 5-year overall survival (OS) and relapse-free survival (RFS) were 80.9% +/- 4.0% and 71.0% +/- 4.0% respectively. The 23 patients relapsed at the median time of 15 months (6 - 70) after CR. Univariate analysis revealed that initial WBC count, first induction regimen, time from induction therapy to CR, type of post-remission therapy and persistent negative RT-PCR in remission were important prognostic factors for long-term survival. Multivariate study demonstrated that only type of post-remission therapy was associated with RFS and OS. It is concluded that the post-remission treatment combining ATRA, As(2)O(3) and chemotherapy would significantly improve the long-term survival of APL patients entering CR(1).
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Arsenicals ; administration & dosage ; Child ; Female ; Follow-Up Studies ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; mortality ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; metabolism ; Oxides ; administration & dosage ; Prognosis ; Proportional Hazards Models ; Remission Induction ; Retrospective Studies ; Survival Analysis ; Survival Rate ; Tretinoin ; administration & dosage

To investigate the related factors affecting the isolation of multipotent non-hematopoietic adult stem cells (MNASCs) from human umbilical cord blood in low serum (2%) condition, the isolation conditions were optimized and the yield of MNASCs was improved. MNASCs from human umbilical cord blood samples were isolated, and the effects of medium component, medium exchange time and initial plating density for isolation of MNASCs were studied. Then, the MNASCs were isolated and cultured in optimal condition, the surface antigen expression and differentiation potential of MNASCs were detected. The result showed that the medium of DMEM/F12 was better than IMDM and DMEM-LG for MNASCs culture in low serum condition. The optimal yield of MNASCs was obtained when mononuclear cells were cultured at a initial plating density of 1 x 10(6) cells/cm2 and the medium was exchanged to remove the nonadherent cells after 72 hours of inoculation. MNASCs isolated and cultured under the above-mentioned conditions maintained a homogenous morphology, high potential ability of expansion and differentiation. It is concluded that culture conditions with low serum defined in this study is optimal for the successful isolation and expansion of umbilical cord blood MNASCs with high numbers for subsequent cellular therapeutic approaches.
Cell Culture Techniques ; methods ; Cell Differentiation ; physiology ; Cell Separation ; methods ; Embryonic Stem Cells ; cytology ; physiology ; Fetal Blood ; cytology ; Humans ; Multipotent Stem Cells ; cytology ; physiology

To compare the expansion efficiency and function of dendritic cells derived from CB-CD34+ cells and MPB-CD34+ cells by using two-step culture method, enriched CB-CD34+ cells or MPB-CD34+ cells with immunoadsorption were primarily cultured in the presence of FL, SCF, TPO, GM-CSF for 10 days, and then further cultured with a combination of GM-CSF, IL-4, TNF-alpha, CD40Ab and PGE2 to induce DC. The DC phenotypes were detected by flow cytometry, the expansion efficiency and cell function were evaluated by mix-lymphocyte reaction (MLR), IL-12 level was detected by using ELISA and the chemotactic function mediated by secondary lymphoid tissue chemokine (SLC) was determined with Transwell plate. The results indicated that after 10 days of expansion, there were no significant difference in the percentage of CD14+CD1a- cells between CB and MPB [(40.48 +/- 16.85)% vs (28.07 +/- 23.19)%, P > 0.05], but the expansion of total cells in CB was higher than that in MPB (388.88 +/- 84.63-fold vs 79.67 +/- 10.32-fold, P < 0.01), so the yield of CD14+CD1a- cells from CB was significantly higher than that from MPB too (189.42 +/- 25.02-fold vs 28.74 +/- 23.27-fold, P < 0.01). The percentage of CD83+ DCs cultured with CD40Ab/PGE2 derived from CB were higher than those cultured with TNF-alpha derived from MPB respectively [(34.52 +/- 11.22)% vs (3.70 +/- 2.27)% and (36.69 +/- 13.36)% vs (7.34 +/- 3.364)% respectively, P < 0.01]. In the same circumstance, the yield of CD83+ DCs derived from CB was much more than that from MPB (198.72 +/- 117.53 times vs 33.95 +/- 6.19 times, P < 0.01). There were no difference in stimulating capacity, IL-12 secretion and migration capacity between DCs derived from CB and MPB. It is concluded that DCs induced from CB-CD34+ cells by two-step culture possess similar functions with that from MPB-CD34+ cells, but the yield of DCs from CB CD34+ cells is much more than that from MPB CD34+ cells.
Antigens, CD34 ; analysis ; Cell Culture Techniques ; methods ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; Hematopoietic Stem Cell Mobilization ; Humans