Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Depression is a serious mental illness with a high incidence. At present, we do not fully understand the specific pathological mechanisms of depression, and the efficacy of drug treatments is very limited. Recent studies have shown that epigenetic changes that occur in specific brain regions may be a key mechanism by which environmental factors to interact with individuals to influence the risk of depression. Therefore, drugs that target epigenetic regulation may become a new direction for the development of antidepressants. Histone deacetylase inhibitors (HDACi) are a class of compounds that inhibit histone deacetylase activity, which has been reported to be associated with depression; this article addresses the use of HDACi in preclinical studies, and their potential therapeutic role and limitations of use in depression.

OBJECTIVETo compare the effects of different types of feeder cells on supporting undifferentiation and high proliferation of human embryonic stem cells (hESC).

METHODShESC were seeded on mouse embryonic fibroblasts (MEF), human marrow stromal cells (hMSC), and human foreskin fibroblasts (hFF), respectively. Colony number, cell quantity after digestion, and survival rate were observed by alkaline phosphatase (AP) staining and Trypan blue, and the biological properties of hESC after 5 passages were observed by immunofluorescence staining.

RESULTSAlthough all the three feeder layers could support the formation of hESC colonies and maintain pluripotency, the morphology of colonies on different feeder layers remarkably varied. The stage-specific embryonic antigen-3 and AP staining were positive on three types of feeders. The number of colonies, number of cells produced, and cell survival rates were significantly higher on MEF than on human feeder cells (P < 0.01). Furthermore, the number of AP-positive colonies and cell quantity were also significantly higher on hMSC than on hFF (P < 0.01).

CONCLUSIONSAll three types of feeder cells are able to support the growth of hMSC, although MEF are more favourable for the proliferation. Two types of human feeder cells lay the foundation for the removal of animal-derived hESC culture system. hMSC is superior to hFF in supporting the proliferation of hESC.

Animals ; Antigens, Tumor-Associated, Carbohydrate ; metabolism ; Coculture Techniques ; methods ; Embryonic Stem Cells ; Feeder Cells ; Fibroblasts ; Humans ; Mice ; Stage-Specific Embryonic Antigens ; metabolism

In this paper the system structure The automatic biochemistry analyzer is a necessary instrument for clinical diagnostics. First of is analyzed. The system problems description and the fundamental principles for dispatch are brought forward. Then this text puts emphasis on the modeling for the automatic biochemistry analyzer control system. The objects model and the communications model are put forward. Finally, the implementation method is designed. It indicates that the system based on the model has good performance.
Autoanalysis ; instrumentation ; methods ; Biochemistry ; instrumentation ; methods ; Equipment Design ; Models, Theoretical

OBJECTIVETo explore the feasibility of transfecting recombinant Sp1 into hypertrophic scar fibroblasts and investigate the proliferation and collagen I, III synthesis in the transfected cells.

METHODSRecombinant human Sp1 was transfected into hypertrophic scar fibroblasts with the karyocyte expressive vector. The expression of Sp1, collagen I, III mRNA was tested by real time PCR. The change of cell proliferation was observed with CCK8 colorimeter.

RESULTSAbout 30% of transfected hypertrophic scar fibroblasts showed green fluorescence positive. The relative expression of Sp1 mRNA in transfected cells, empty-vector cell or untransfected cells group was 5.26 +/- 0.76, 1.08 +/- 0.18, 1.09 +/- 0.15, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group (P <0.01, n = 5). Expression of collagen I, III mRNA was 2.49 +/- 0.40 and 1.88 +/- 0.30 in transfected cells, 0.96 +/- 0.18 and 0.95 +/- 0.18 in empty-vector cell, and 0.97 +/- 0.15 and 0.93 +/- 0.13 in untransfected cells, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group (P < 0.01, n = 5).

CONCLUSIONSThe hypertrophic scar fibroblasts could be as the target cells of Sp1 gene transfection. Sp1 gene may play an important role in abnormal collagen metabolism in hypertrophic scar.

Cell Proliferation ; Cells, Cultured ; Cicatrix, Hypertrophic ; genetics ; metabolism ; pathology ; Collagen ; metabolism ; Escherichia coli ; genetics ; Fibroblasts ; metabolism ; pathology ; Humans ; RNA, Messenger ; genetics ; Recombinant Proteins ; genetics ; Skin ; metabolism ; Sp1 Transcription Factor ; genetics ; Transfection

OBJECTIVETo study the alteration of thymus matrix lymphocyte generator (TSLP) and change of the Th factor in the course of disease development, and to analyze the curative effect of inhalation of Vitamin A (VA) with corticosteroid for the treatment of asthmatic pneumonia.

METHODSAsthmatic pneumonia models were prepared by challenging rats with inhalation of ovalbumin for 4 weeks, and rested for 1 week. The treatment with VA and corticosteroid inhalation for 1 week was followed. The rat thymus and lung specimen were examen by histochemical and immunofluorescence staining.

RESULTSAfter 4 - 5 weeks of stimulation, there were more TSLP-positive cells and alveolar macrophages (AM) found in thymus and lung tissue of asthmatic group, the cell proliferation in spleen and thymus was obvious, and blood Th factors elevated. The inflammation within the lung tissue aggravated gradually. In VA group, the expression of TSLP and Th2 factors were all lowered at the 4th week. The TSLP expression slightly increased at the 5th week, and the cell proliferation within T-cell zone of spleen and thymus was strong at first and weakened later. Alveolar microphages (AM) increased significantly and the inflammation in the lung subsided gradually at the 5th week. In the hormone group, TSLP and Th2 factors expression in both thymus and lung were decreased at the 5th week, while the cell proliferation in thymus and lung was gradually increased. The quantity of AM was decreased, whereas the inflammation of the lung was increased gradually at the 5th week.

CONCLUSIONDuring asthmatic period elevated TSLP expression was accompanied by Th2 type responses while VA and corticosteroid both suppressed TSLP and Th2 factors expression. VA alone promoted T lymphocyte proliferation as well as the antigen elimination function by AM, after ceasing the usage, the lung inflammation abated gradually. In contrast, after ceasing the use of corticosteroid, inflammation aggravated.

Administration, Inhalation ; Adrenal Cortex Hormones ; administration & dosage ; therapeutic use ; Animals ; Asthma ; complications ; Beclomethasone ; administration & dosage ; therapeutic use ; Cytokines ; metabolism ; Pneumonia ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vitamin A ; administration & dosage ; therapeutic use

OBJECTIVETo investigate the fine needle aspiration cytology (FNAC) features and differential diagnosis of eyelid sebaceous gland carcinoma.

METHODSFour cases of eyelid sebaceous gland carcinoma diagnosed by FNAC were reported and confirmed by biopsy. Three of the cases were in early stages with tumor sizes smaller than 10 mm in diameter and without metastasis. The smears were stained by routine H & E and SudanIII methods. The cytologic findings were described and compared to corresponding histological features, and moreover, compared to chalazion, pilomatrixoma and eyelid basal cell carcinoma.

RESULTSNeither hemorrhage nor infection were found after the examination. Abundant cells were observed in the sebaceous carcinoma FNAC smears. Two types of tumor cells were found: one showed tumor cells differentiating toward sebaceous gland, with large pale cells and vacuolated cytoplasm, the other demonstrated poorly-differentiated cell with dark and irregular nuclei. Numerous vacuoles with inequality of size were found in cytoplasm or in background in all four cases, and the SudanIII stain showed that these vacuoles contained lipid. Some smears demonstrated cells with basaloid, fusiform or squamous features, corresponding to various histopathological types. In contrast, smears of chalazion displayed inflammatory granuloma, containing several types of inflammatory cells without malignant cells. Smears of pilomatrixoma were cellular with three cell populations, which included bland sheets of basaloid cells, nucleated basophilic cells and anucleated keratinized "ghost cells", along with calcific debris. The smears of basal cell carcinoma were typically less cellular, more tightly cohesive and had smaller clusters of uniform hyperchromatic basaloid cells without vacuolization in cytoplasm or background. Overall, the cytological features of eyelid sebaceous carcinoma were distinct from those of chalazion, pilomatricoma and basal cell carcinoma.

CONCLUSIONSFNAC is a safe and effective approach for the diagnosis of eyelid sebaceous carcinoma and lipid stain is useful in differential diagnosis. The application of FNAC may be important in reaching an early diagnosis and initial treatment of eyelid nodule.

Adult ; Aged ; Biopsy, Needle ; Diagnosis, Differential ; Eyelid Neoplasms ; diagnosis ; pathology ; Female ; Humans ; Male ; Middle Aged ; Sebaceous Gland Neoplasms ; diagnosis ; pathology

This study was purposed to explore the inhibitory effect of Curcumin on growth of retinoic acid-resistant acute promyelocytic leukemia (APL) cells and its mechanism. The NB4-R1, an APL cell line resistant to retinoic acid, was used as a model. The growth level of NB4-R1 was detected by MTT assay, the morphologic features of cells were observed by light microscopy, the mitochondrial transmembrane potential was determined by flow cytometry, the expressions of apoptosis-related proteins procaspase 3, caspase 3, PARP and BCL-XL were measured by Western blot. The results indicated that the sensitivity of NB4-R1 to Curcumin was consistent with NB4 though NB4-R1 was resistant to retinoic acid, Curcumin displayed inhibitory effect on growth of NB4-R1 in time-and concentration-dependent manners. The morphologic observation showed existence of apoptotic bodies in NB4-R1 cells treated with 20 micromol/L of Curcumin. The flow cytometry indicated that the mitochondrial transmembrane potential in NB4-R1 cells treated with 20 micromol/L of Curcumin obviously decreased. The Western blot detection revealed that expressions of pro-caspase 3 and BCL-XL were down-regulated, expressions of caspase 3 and sheared PAPP were up-regulated in NB4-R1 cells treated with 20 micromol/L of Curcumin. It is concluded that the Curcumin can inhibit the growth and induce the apoptosis of NB4-R1.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Curcumin ; pharmacology ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Tretinoin ; pharmacology ; bcl-X Protein ; metabolism

Objective To observe the effect of intravenous nutrition on blood serum triglyceride (TG),humoral immunity and cellular immunity function in premature infants.Methods Sixty premature infants were randomly divided into 3 groups:amino acid group (group A),amino acid plus medium-long chain fatty group (group B) and amino acid plus long chain fatty group (group C).Amino acid and the fatty were used on them from 24 hours after their birth,started from 1.0 g/(kg?d),increased progressively 0.5 g/(kg?d) until 3.0 g/(kg?d),totally 7 days.TG and immunoglobulin IgA,IgM,IgG,complement C3,C4 and T-lymphocyte subsets CD3,CD4,CD4/CD8 and NK cell were checked in the first day before treatment and the ninth day after treatment.Results Compared with before treatment,TG of 3 groups were elevated(Pa0.05).Compared with group A and B,NK cell in group C were decreased obviously(Pa

Objective To investigate the effects of 3 different ventilation methods,including conventional mechanical ventilation (CMV),high frequency oscillatory ventilation(HFOV) and partial liquid ventilation (PLV),on the changes of inflammatory factors and pulmonary surfactant associated protein A (SP-A) in bronchoalveolar lavage fluid (BALF) in newborn piglets with acute lung injury(ALI).Methods Twenty-four newborn piglets,no more than 3 days old,were enrolled.After ALI made with saline lavage(38 ℃,35 mL/kg),newborn piglets were randomly assigned to 4 groups:control group (n =6,no ventilation),CMV group(n =6),HFOV group(n =6),and PLV group(n =6).Piglets were sacrificed after being ventilated for 24 h.Tumor necrosis factor-α (TNF-α),interleukin-8 (IL-8),interleukin-1 (IL-1) and SP-A in BALF were measured quantitatively by using enzyme-linked immunosorbent assay.Results In 3 groups using different ventilation methods,the population mean of TNF-o,IL-8,IL-1 and SP-A were statistically different (all P =0.000).SP-A in PLV group and HFOV group were higher than that in CMV group (all P ＜ 0.05),while IL-8,IL-1 and TNF-α in PLV group were lower than those in CMV group (all P ＜ 0.05),IL-8 and TNF-α in PLV group were lower than those in HFOV group (all P ＜ 0.05),IL-8 and TNF-α in HFOV group were lower than those in CMV group (all P ＜ 0.05).Conclusions Pulmonary inflammatory reaction was different in 3 ventilation groups.Compared with CMV and HFOV,PLV attenuated inflammatory reaction,so it could increase the expression of SP-A and decrease the degradation of SP-A.