Adenocarcinoma ; pathology ; Adult ; Aged ; Female ; Humans ; Lasers ; Male ; Middle Aged ; Neuroendocrine Cells ; pathology ; Polymerase Chain Reaction ; methods ; Stomach Neoplasms ; pathology

Objective To observe non-displaceable binding potential (BPND) changes of striatal dopamine D2 receptors(SDDR) in patients with first-episode major depressive disorder (MDD) using 11C-Raclopride PET/CT,and to analyze the relationship between BPND and Hamilton rating scale for depression (HAM-D).Methods From December 2014 to December 2015,patients with first-episode MDD and age/gender-matched healthy controls underwent brain MRI and 11C-Raclopride PET/CT in this prospective study.BPND of bilateral SDDR was calculated by molecular imaging and kinetic analysis toolbox (MIAKAT).BPND changes of bilateral SDDR and their relationship with HAM-D score were analyzed.Paired t test,two-sample t test and Pearson correlation analysis were used.Results A total of 20 MDD patients (8 males,12 females,average age: (32.80±9.76) years) and 20 healthy controls (9 males,11 females,average age:(29.25±6.93) years) were enrolled in this study.The 11C-Raclopride uptake in brain tissue of the MDD group and control group were mainly distributed in bilateral striatum,and very few 11C-Raclopride was distributed in bilateral cerebral cortex and cerebellum.In MDD group,the BPND level of bilateral SDDR had no statistical differences(t values: 0.69,0.35,both P>0.05),and similar results were found in the control group(t values: 0.28,0.24,both P>0.05).Compared with the control group,however,the MDD group had lower BPND level of bilateral SDDR(t values: 3.13-4.41,all P<0.05).The BPND of bilateral caudate nucleus and/or putamen D2 receptors was correlated with HAM-D total score,anxiety/somatization factor score,cognitive impairment factor score,retardation factor score and sleep disturbance factor score(r values: from-0.688 to-0.453,all P<0.05).Conclusions The binding potential of SDDR in patients with first-episode MDD is declined,and the BPND level of SDDR is correlated with symptoms of depression.The abnormality of SDDR may be an important molecular mechanism of the abnormality of midbrain-striatal dopamine reward circuits in MDD patients.

OBJECTIVETo observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate.

METHODSImmature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay.

RESULTSThe DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading.

CONCLUSIONBoth methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; biosynthesis ; immunology ; Carcinoembryonic Antigen ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; therapy ; Dendritic Cells ; immunology ; metabolism ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Interleukin-12 ; metabolism ; Transfection

OBJECTIVENeonatal hypoxic-ischemic brain damage (HIBD) causes acute death and chronic nervous system sequelae in newborn infants and children. Whereas there have been no specific treatment towards it up to now. Studies have shown that bone marrow mesenchymal stem cells (MSCs) have the therapeutic potential in many nervous system diseases and the authors previously found that retinoid acid (RA), which plays an important role in brain development, could enhance the neural differentiation of rat MSCs (rMSCs) in vitro. This study aimed to examine effects of rMSCs and RA-preinduced rMSC on learning and memory functional recovery after HIBD in neonatal rats in order to explore a new treatment strategy for clinical application, and explore the mechanism of action of rMSCs.

METHODSRat MSCs were isolated and purified from the whole bone marrow of juvenile Wistar rats by removing the non-adherent cells in primary and passage cultures. Neonatal hypoxic-ischemic brain damage rat models were built according to the methods described by Rice: the right carotid artery of 7-day-postnatal Wistar rats was ligated under anesthesia, and then the rats were exposed to 8% - 9% O2 in a container. At 5 days after hypoxia-ischemia, the HIBD neonatal rats were randomly divided into 3 groups and respectively transplanted with saline, BrdU marked rMSCs (1 - 2 x 10(5)) or RA-preinduced rMSCs (1 - 2 x 10(5)) into their lateral cerebral ventricle. Immunohistochemistry for nestin, neuron-specific enolase (NSE), neurofilament protein-heavy chain (NF-H) and glial fibrillary acidic protein (GFAP) were used to identify cells derived from rMSCs at 14 days and 42 days after transplantation. Shuttle box test was performed to evaluate the condition of learning and memory functional recovery when animals were 7 weeks old. Neurotrophin and receptors cDNA microarray were also employed at 14 days after transplantation to investigate the underlying action mechanisms of rMSCs treatment. Real-time PCR was used to confirm some of the remarkably changed genes.

RESULTS(1) The neonatal rat model of HIBD was successfully established. (2) Immunohistochemistry showed rMSCs-derived cells survived, migrated into the hypoxic-ischemic brain tissue and a few of them expressed protein characteristic of neurons and astrocytes (NF-H and GFAP) in RA-preinduced group 14 days and 42 days after transplantation, while no positive expression of nestin and NSE were detected. (3) The shuttle box test showed that the average learning times in rats transplanted with saline, rMSC and RA-preinduced rMSCs were (94.10 +/- 38.18), (74.60 +/- 29.21) and (47.90 +/- 21.13), respectively. The difference between the former two was not significant (P > 0.05), while the latter one exhibited significant improvement (P < 0.05). (4) The cDNA microarray analysis showed that compared with normal control group, IL-6, Fas and BDNF genes of the saline control group significantly up-regulated (the ratios of the three genes were 11.4, 2.4 and 6.6 respectively). Compared with saline group, the three genes in rMSC group were down-regulated (the ratios were all 0.1), while the levels of IL-6 and Fas genes (the ratios were 0.3 and 0.4 respectively) in RA-preinduced rMSCs group were higher than rMSCs group after down-regulating, but the level of BDNF remained at the saline group level. Real-time PCR analysis suggested that the results of IL-6 and Fas genes were at equal level with microarray results on the whole, while the level of BDNF gene in RA-preinduced rMSC group was significantly down-regulated (with ratio of 0.34), but higher than rMSCs group (the ratio was 0.25) as well.

CONCLUSIONTransplantation of rMSC and RA-preinduced rMSCs into lateral cerebral ventricle can improve learning and memory functional recovery after HIBD in neonatal rats, especially RA-preinduced rMSCs. Regulating the levels of IL-6, Fas and BDNF in the brain to maintain at reasonable levels may be the mechanism.

Animals ; Animals, Newborn ; Cell Differentiation ; Cells, Cultured ; Hypoxia-Ischemia, Brain ; psychology ; therapy ; Memory ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent.

METHODSHuman DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test.

RESULTSAn exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection.

CONCLUSIONMicrobubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.

Adaptor Proteins, Signal Transducing ; genetics ; Cells, Cultured ; Contrast Media ; administration & dosage ; pharmacology ; Cytoskeletal Proteins ; genetics ; Dendritic Cells ; drug effects ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; Microbubbles ; Plasmids ; Transfection ; Ultrasonics

Objective:To prepare 68Ga-2-(4, 7-bis(carboxymethyl)-1, 4, 7-triazonan-1-yl)pentanedioic acid (NODAGA)-YHWYGYTPQNVI (GE11) and evaluate its feasibility of PET imaging for pancreatic cancer. Methods:GE11 peptide was conjugated with NODAGA and then labeled with 68Ga. The labeling yield, radiochemical purity, hydrophilicity, stability and specificity in vitro were determined. Human pancreatic cancer BxPC3 nude mice models ( n=9) were established. MicroPET imaging was then obtained after 30 and 90 min, and mice were sacrificed at 90 min to acquire the radioactivity distribution of main organs and tumors. Pair t test was used to analyze the data. Results:The labeling yield was (73.5±5.4)% and radiochemical purity was more than 98%. After incubation 120 min in mouse serum at 37 ℃, radiochemical purity was more than 92%. The uptake was specific in BxPC3 cell lines. MicroPET images showed that 68Ga-NODAGA-GE11 could accumulate quickly in tumor. Value of tumor uptake was significantly higher than that of normal pancreas at 90 min ((1.38±0.25) vs (0.49±0.07) %ID/g; t=12.67, P<0.05), and the radio-uptake of blood, muscle and bone was lower than that of tumor. Conclusions:68Ga-NODAGA-GE11 is easy to be prepared with high radiochemical purity and good stability, and can specifically target BxPC3 xenograft tumor. However, due to the high uptake in the kidneys and liver, the value of 68Ga-NODAGA-GE11 in PET imaging for pancreatic tumor needs further study.

OBJECTIVETo construct a recombinant adenovirus vector carrying soluble extracellular region of tumor necrosis factor alpha receptor I-IgGFc (sTNFRI-IgGFc) and express the fusion protein in human bronchial epithelial HBE135-E6E7 cells.

METHODSsTNFRI-IgGFc fusion gene was subcloned into the adenovirus shuttle plasmid pDC316, which was co-transfected with helper plasmid pBHGloxPE1,3Cre into HEK293 cells. The recombinant adenovirus (Ad-sTNFRI-IgGFc) was generated by homologous recombination of the 2 plasmids in HEK293 cells. After identification with PCR, Ad-sTNFRI-IgGFc was amplified and purified, and its titer measured using TCID50 assay. The transcription and expression of sTNFRI-IgGFc gene in the transfected HBE135-E6E7 were detected by RT-PCR and immunohistochemistry.

RESULTSAd-sTNFRI-IgGFc was successfully constructed with a viral titer of 3 x 10(10) TCID50/ml. The expression of sTNFRI-IgGFc mRNA and protein was confirmed in the transfected HBE135-E6E7 cells.

CONCLUSIONThe constructed Ad-sTNFRI-IgGFc can effectively infect HBE135-E6E7 cells for efficient expression of sTNFRI-IgGFc protein, which antagonizes the cytolytic effect of TNFalpha in L929 cells, suggesting the potential of adenovirus expressing sTNFRI-IgGFc for local treatment of asthma.

Adenoviridae ; genetics ; Bronchi ; cytology ; Epithelial Cells ; cytology ; metabolism ; Genetic Vectors ; genetics ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor, Type I ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo assess the value of (18)F-fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography ((18)F-FDG PET-CT) in ultrasound-guided local ablation of malignant liver tumors.

METHODSThirteen patients with 35 local residual tumor foci following previous tumor ablation underwent (18)F-FDG PET-CT and ultrasound-guided local ablation with intratumoral alcohol injection.

RESULTSAfter the second local ablation guided by (18)F-FDG PET-CT and ultrasound, radioactive defects were detected in the corresponding location in 31 of the 35 residual foci, and after the third local ablation, the other 4 foci also showed radioactive defects.

CONCLUSION(18)F-FDG PET-CT can sensitively and accurately identify tissue necrosis and residual tumors, and serves as an excellent approach for ultrasound-guided local ablation of local residual tumors.

Ablation Techniques ; adverse effects ; Adult ; Aged ; Female ; Fluorodeoxyglucose F18 ; Humans ; Liver Neoplasms ; diagnosis ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Positron-Emission Tomography ; Postoperative Complications ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

OBJECTIVETo study the feasibility of transfecting breast cancer BA46 gene into dendritic cells (DCs) using adeno-associated virus (AAV) to induce specific cellular immunity.

METHODSMononuclear cells (DC precursor) were isolated from the peripheral blood of healthy donors by density gradient centrifugation and infected with rAAV/BA46/Neo virus stock (transfection group) or pulsed with 293 cell lysate (control group). In both groups, maturation of the DC precursor was induced by granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha(TNF-alpha). On day 7, the DCs were collected and mixed with T cells at the ratio of 1 to 20 to induce cytotoxic T lymphocytes (CTL). The capacity of the DCs in stimulating T lymphocyte proliferation was assessed using (3)H-thymidine incorporation assay. The expressions of interferon-gamma (IFN-gamma), IL-4, CD4, CD8, CD25 and CD69 in the CTLs were analyzed with cytometry, and the cytotoxicity of the CTLs was evaluated with (51)Cr-release assay using BA46-positive breast cancer cell line Hs578T as the target.

RESULTSThe DCs transfected with BA46 gene exhibited potent capacity to stimulate T lymphocyte proliferation. The CTL population induced by the transfected DCs expressed high levels of CD8, CD69 and IFN-gamma, and showed strong cytotoxicity against BA46-positive breast cancer cell line Hs578T, which was BA46 antigen-specific and MHC-limited.

CONCLUSIONThe success in BA46 gene transfer in the DCs that induce specific cellular immunity provides the experimental basis for breast cancer immunotherapy using genetically modified cells.

Antigens, Surface ; genetics ; metabolism ; Breast Neoplasms ; genetics ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; metabolism ; Dependovirus ; genetics ; metabolism ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Immunity, Cellular ; Immunotherapy ; Interleukin-4 ; pharmacology ; Milk Proteins ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection

Objective To analysis the influencing factors on underweight and stunting among children aged 0-3 years in rural areas from ten provinces in China.Methods Children under study were identified by multi-stage stratified cluster from rural areas of ten provinces in China.The ascertainment methods mainly included questionnaire and anthropometric measurements.Results There were 58 926 children under investigation,with 50.91％ were boys.The overall rates on underweight and stunting were 5.05％ and 10.49％ respectively.The rate in the 6 month-olds (1.97％,3.79％ ) was the lowest,while the highest were in the 24 month-olds (7.80％) and the 36 month-olds (16.83％).Age,sex,birth weight,gestational weeks as well as maternal education and fathers' schooling were factors significantly related to childhood underweight and stunting (P＜0.0001).Conclusion The status of underweight and stunting among children aged 0-3 years in rural areas was impressive,with birth weight was the key factor influencing the growth of children.Perinatal health care should be improved to promote the growth of children.