Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Neovascularization, Pathologic ; pathology ; Tryptases ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

Objective To study the effects of anti -cancer drug NSC319726 on the gene expression of ovarian cancer.Methods The data that NSC319726 acted on the ovarian cancer cell lines TOV112D was down-loaded from the GEO database and then the bioinformatical analysis was conducted .Results NSC319726 in-creased 246 genes including MMP3,CXCR4 et al and decreased 198 genes such as PBX1 in ovarian cancer cell lines.GO analysis indicated that six GO items were related to metabolism .Pathway analysis revealed that NSC319726 could affect the circadian clock and MMP signaling pathway .Conclusion NSC319726 could affect the expression of multiple functional genes in ovarian cancer cell lines and might be a potential drug targeting to the precursor-B cell acute lymphoblastic leukemia .Meanwhile , it should be noted that NSC 319726 may have effects on the tumor metastasis and human sleep .Therefore,further research needs to be conducted before the clinical use .

According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

Objective To investigate the risk factors for postoperative arytenoid dislocation.
Methods From September 2003 to August 2013, the records of 16 patients with a history of postoperative arytenoid dislocation were reviewed. Patients matched in terms of date and type of procedures were chosen as the controls (n=16). Recorded data for all patients were demographics, smoking status, alcoholic status, preoperative physical status, airway evaluation, intubation procedures, preoperative laboratory test results, anesthetic consumption and intensive care unit stay. For arytenoid dislocation cases, we further analyzed the incidences of the left and right arytenoid dislocation, and the outcomes of surgical repair and conservative treatment. Categorical variables were presented as frequencies and percentages, and were compared using the chi-squared test. Continuous variables were expressed as means±SD and compared using the Student’s unpaired t-test. To determine the predictors of arytenoid dislocation, a logistic regression model was used for multivariate analysis.
Results Sixteen patients with postoperative arytenoid dislocation were enrolled, with a median age of 52 years. Most postoperative arytenoid dislocation patients (15/16, 93.75%) received surgical repair, except one patient who recovered after conservative treatment. None of the postoperative arytenoid dislocation patients were smokers. Red blood cell (P=0.044) and hemoglobin (P=0.031) levels were significantly lower among arytenoid dislocation cases compared with the controls.
Conclusions Non-smoking and anemic patients may be susceptible to postoperative arytenoid dislocation. However, neither of them was independent risk factor for postoperative arytenoid dislocation.

Objective To evaluate the changes in epithelial thickness profile and its relationship with diopter following small incision lenticule extraction (SMILE) for moderate and high myopia.Methods Together 46 myopia or myopic astigmatism (92 eyes) who underwent SMILE were included under the informed consent from January 2016 to March 2017 and were decided into 2 groups according to the diopter:moderate myopia group (58 eyes)and high myopia group (34 eyes).Epithelial thickness profile was measured using spectral-domain optical coherence tomography at different corneal zones (0-2 mm,＞ 2-5 mm and ＞ 5-6 mm cornea) preoperatively before surgery and 1 month and 6 months after surgery for observing the changes in epithelial thickness and its correction with diopter.Results The mean epithelial thickness in the central zone was (55.68 ± 3.61) μm before surgery,and,6 months after surgery,it was thickened by (3.85-±3.99),(3.46 ±3.29) and (2.85 ±3.18) μm in the 0-2 mm,＞2-5 mum and ＞5-6 mm cornea respectively,and the difference was statistically significant among the three zones (P ＜ 0.01).After surgery for 6 months,the epithelial thickness in the high myopia group was thickened more obviously compared with the moderate myopia group (t =1.440,P =-0.047).And no correlation was found between changes in the epithelial thickness at 0-2 mm cornea and diopter after surgery(moderate myopia group:r =0.219,P=0.633;high myopia group:r =0.197,P =0.585).Conclusion Significant epithelial thickening was observed after SMILE,presenting the thickened epithelium.The higher the diopter was,the more thickened the epithelium was.The epithelial changes does not appear to affect the diopter after SMILE.

OBJECTIVE Programmed death ligand-1 (PD-L1) and indoleamine 2, 3-dioxygenase 1 (IDO1) are immune checkpoints which can be induced by interferon-γ(IFN-γ) in the tumor microenvironment, leading to immune escape of tumors. Myricetin (MY) is a flavonoid distributed in many edible and medicinal plants. The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells. METHODS Expressions of PD-L1 and major histocompatibility complex-I (MHC-I) were evaluated by flow cytometry and Western blotting, and the expression of IDO1 was measured by Western blotting. qRT-PCR was used to detect their mRNA levels. The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1. Molecular docking analysis, Western blotting and immunofluorescence were used for mechanism study. RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells, while didn't show obvious effect on the expression of MHC-I. In addition, MY restored the survival, proliferation, CD69 expression and interleukin-2 (IL-2) secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system. Mechanistically, IFN-γ up-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis, which was targeted and inhibited by MY. CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression, supporting the potential of MY in anti-tumor immunotherapy.

This study was aimed to analyze the biological characteristics of rabbit bone marrow mesenchymal stem cells (rBM-MSCs) and their response to different growth factors. Rabbit BM-MSCs were separated from bone marrow mononuclear cells by using adherent cultivation. Biological characteristics were investigated by optical and electron microscopy. Immunophenotype of rBM-MSCs was measured by flow cytometry. The expression of collagen was detected by RT-PCR. Differentiation potential was identified by specific staining and RT-PCR. The response of rBM-MSCs to IL-1, 3, 8 and HGF with different concentrations were tested by MTT. The results showed that the rBM-MSCs gave rise to a population of adherent cells characterized by the presence of a predominant cell type with a typical fibroblast-like morphology and could be cultured for over 15 passages. CD44 was highly expressed on F5 rBM-MSCs (32%) and CD45 was lowly expressed (4.7%). Type I collagen was highly expressed, while type II collagen was lowly expressed and type X collagen was not detected on rBM-MSCs using RT-PCR method. In various conditions inducting differentiation, rBM-MSCs could differentiate into the osteoblast, chondrocyte, adipocyte and neuron-like cells. The rBM-MSCs were sensitive to IL-3, even low concentration (10 ng/ml) of IL-3 could promote the proliferation of rBM-MSCs effectively (>32%, P < 0.01), whereas high concentration IL-3 inhibited it significantly. It is concluded that rabbit BM-MSCs were successfully isolated and culture-expanded. The biological characteristics of rabbit BM-MSCs are similar to those of human and rhesus BM-MSCs. IL-3 with low concentration can promote the proliferation of rBM-MSCs effectively, but high concentration of IL-3 can inhibit their proliferation.
Animals ; Bone Marrow Cells ; cytology ; physiology ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Cytokines ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; physiology ; Rabbits ; Recombinant Proteins

BACKGROUNDTo clarify the features of gene variation among epidemic strains of human rotavirus NSP4 in China.

METHODSSP4 genes from 27 epidemic strains of human rotavirus isolated in different area of China in recent years were amplified with RT-PCR, the resulted cDNAs were cloned and sequenced. The sequences of full length cDNAs were compared with 10 rotavirus NSP4 sequences available in the GenBank using the Clustal x 1.8 TreeView32 and DNA Star softwares. The G serotype of VP7 was analysed by PCR.

RESULTSThe homology of the amino acid among the 27 rotavirus strains isolated in China was 81.7%-99.4%. Based on the variation of amino acid sequence, the virus strains can be divided into two groups, represented by Wa and KUN with the homology of 92.0%-99.4% and 92.0%-98.9% within each group, respectively. The diversity between the two groups were 16.6%-21.0%. The Wa group could further be separated into three subgroups, according to the diversity between those strains and the characterization in the highly variable domain. The association between VP7 serotype and NSP4 genotype was not strong.

CONCLUSIONSThe NSP4 gene of human rotavirus epidemic strains in China can be divided into Wa and KUN two groups, Wa group is the main group and contains three subgroups possessing characteristic amino acid sites. Samples isolated in the same year but not in the same area shared higher homology.

Antigens, Viral ; Capsid Proteins ; genetics ; DNA, Complementary ; analysis ; DNA, Viral ; analysis ; Diarrhea ; virology ; Genetic Variation ; Genotype ; Glycoproteins ; genetics ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Toxins, Biological ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo investigate the effect of hTERT antisense oligodeoxynucleotide (ASODN) on the oncogenicity and the inductive apoptosis of HL-60 cells.

METHODSApoptosis of HL-60 cells was detected by flow cytometry (FCM) and agarose gel electrophoresis. Both treated and untreated HL-60 cells were collected and transplanted into 5 BALB/c nude mice respectively, the formation of transplanted neoplasm and its morphologic change were observed. After the transplanted neoplasms were uniform with the ameliorated method in another 10 BALB/c nude mice, they were divided into 2 groups and injected ASODN and PBS into the neoplasm respectively. Seven days later, the tumor were measured, its morphology were observed, and the apoptotic cells were detected with a TUNEL kit.

RESULTSAfter 72 h treatment there were DNA ladders and early apoptosis peak in hTERT ASODN treated HL-60 cells but was none in SODN treated and blank control cells. In tumor formation experiment, neoplasms were formed in ASODN treated group at 16-17 d and untreated group at 12-13 d. Neoplasm was formed in 2 of 5 ASODN treated mice and 4 of 5 untreated mice respectively. In untreated mice tumor tissues were rich in blood vasa and stromal tissue compared with that in ASODN treated mice. In tumor therapy experiment, before treatment, there was no difference in the average neoplasm physical volume between ASODN treated group [(100.9 +/- 24.6) mm3] and PBS treated group [(98.4 +/- 23.1) mm3] (P > 0.05). After treatment, the neoplasm volume in ASODN treated group [(422.7 +/- 326.4) mm3] was smaller than that in PBS treated group [(786.4 +/- 357.6) mm3] (P < 0.05). Histologically, there were many apoptosis cells in ASODN treated group, but was seldom seen in PBS treated group. The TUNEL positive cells in ASODN treated group were much more than that in PBS treated group (P < 0.05).

CONCLUSIONThe hTERT ASODN induces apoptosis of HL-60 cells in vitro, reduces the tumor formation in BALB/c nude mice and inhibits the growth of the transplanted neoplasm.

Animals ; Apoptosis ; drug effects ; HL-60 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Telomerase ; genetics ; Transfection ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the influence of the prostaglandin E2 on the proliferation of the melanocytes in the full-thickness skin graft.

METHODSSixty-eight guinea-pigs were divided into experimental-1 group (skin graft), experimental-2 group (skin graft + diclofenac), and control groups. After the full-thickness skin graft, the dynamic changes of the prostaglandin E2 were measured and the proliferation of the melanocyte with its density was also evaluated by using histochemical and autoradiographic methods.

RESULTSIn the experimental-1 group, the content of PGE2 was increasing in seven days after the operation, continued to the one month, and then returned to the base level. The labelling indices of 3H-MC-TdR of the group was also increasing postoperatively between the second day and the fourteenth day, and reach a second peak after one month, then came to the normal level. The density of the melanocytes was decreasing rapidly 3 days after the surgery, then began to increase and exceeded over the normal level 21 days after the operation. However, in the experimental-2 group, the content of PGE2 decreased in two days after the surgery, and then showed the inclination similar to the experimental-1 group with the different points in narrower range. The number of melanocytes labelled by 3H-TdR began to increase at the first day after the surgery, which appeared earlier than the experimental-1 group and was similar in the changing tendency with a less extent. The density of MC showed the similar tendency to the experimental-1 group in a narrower changing range with both of increasing and decreasing. The density of the MC was much lower in 21 after the operation than the experimental-1 group and normal control group.

CONCLUSIONThe increased PGE2 in the earlier stage of the skin grafting could enhance the inflammatory reaction to the tissue, as well as the melanocytes. It may stimulate the proliferation of the MC with the result of increasing their density. The use of the diclofenac might reduce the inflammation and suppress the proliferation of melanocytes, and result in the skin with light color due to decreasing the number of MC in the epidermis of the graft.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Cell Count ; Cell Proliferation ; Diclofenac ; pharmacology ; Dinoprostone ; metabolism ; Epidermis ; Guinea Pigs ; Melanocytes ; cytology ; Skin ; Skin Transplantation ; Time Factors