Bone Neoplasms ; Diagnosis, Differential ; Humans ; Osteoblastoma/surgery*

Salivary glands are important organs in the oral and maxillofacial region. Environment and genetic factors may cause salivary gland tumors or non-neoplastic diseases, but the mechanisms of those diseases are still unclear. One of the important reasons is the short of researching media and model. As a new technique and research model, organoids have been widely used in the research of various diseases. Organoid culture plays a bridging role between two-dimensional cell culture and living animal models, and it is also the most promising translational research model that could connect the clinical research to basic research. This review will discuss the recent development of organoid techniques in the culture of normal salivary glands and salivary gland tumors, also their applications and challenges in tissue engineering, etiological research, and tumor therapy.
Animals ; Cell Culture Techniques ; Organoids ; Salivary Gland Neoplasms ; Salivary Glands ; Tissue Engineering

OBJECTIVE: To explore the feasibility of preparing gastric floating formulations by fused de-position modeling (FDM) 3D printing technology, to evaluate the in vitro properties of the prepared FDM 3D printed gastric floating formulations, and to compare the influence of different external shapes of the formulation with their in vitro properties. METHODS: Verapamil hydrochloride and polyvinyl alcohol (PVA) were used as the model drug and the excipient, respectively. The capsule-shaped and hemisphere-shaped gastric floating formulations were then prepared by FDM 3D printing. The infill percentages were 15%, the layer heights were 0.2 mm, and the roof or floor thicknesses were 0.8 mm for both the 3D printed formulations, while the number of shells was 3 and 4 for capsule-shaped and hemisphere-shaped formulation, respectively. Scanning electron microscopy (SEM) was used to observe the morpho-logy of the surface and cross section of the formulations. Gravimetric method was adopted to measure the weights of the formulations. Texture analyzer was employed to evaluate the hardness of the formulations. High performance liquid chromatography method was used to determine the drug contents of the formulations. The in vitro floating and drug release behavior of the formulations were also characterized. RESULTS: SEM showed that the appearance of the FDM 3D printed gastric floating formulations were both intact and free from defects with the filling structure which was consistent with the design. The weight variations of the two formulations were relatively low, indicating a high reproducibility of the 3D printing fabrication. Above 800.0 N of hardness was obtained in two mutually perpendicular directions for the two formulations. The drug contents of the two formulations approached to 100%, showing no drug loss during the 3D printing process. The two formulations floated in vitro without any lag time, and the in vitro floating time of the capsule-shaped and hemisphere-shaped formulation were (3.97±0.41) h and (4.48±0.21) h, respectively. The in vitro release of the two formulations was significantly slower than that of the commercially available immediate-release tablets. CONCLUSION The capsule-shaped and hemisphere-shaped verapamil hydrochloride gastric floating formulations were prepared by FDM 3D printing technology successfully. Only the floating time was found to be influenced by the external shape of the 3D printed formulations in this study.
Drug Liberation ; Excipients ; Printing, Three-Dimensional ; Reproducibility of Results ; Tablets

The transplants of the two-year-old Glycyrrhiza uralensis were subjected to four concentration of brassinolide (BR 0.1, 0.4, 0.7, 1.0 mg•L⁻¹) in July. The morphological characters ( plant height, stem diameter, nodes number, internode length and root length , root thick, root fresh weight and root dry weight ) were measured and seven kinds of chemical constituents (glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin, liquiritin apioside, isoliquiritin apioside) were determined by HPLC with the aim of increasing sinter output and improving quality of G. uralensis. Then the long-term dynamic changes of these morphological characters and chemical compositions' content were analyzed. The results showed that morphological characters of plant height, stem diameter, root length , root thick, root fresh weight and root dry weight increased remarkably with the 0.7 mg•L⁻¹ BR stimulating 2 months later,the increase rates were： 15.09%,6.15%,16.52%,8.46%,21.90%,29.41%, respectively. The content of glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, liquiritin apioside, isoliquiritin apioside were increased 20.16%,45.31%,53.56%,27.66%,23.54%,8.46% with the 0.7 mg•L⁻¹ BR stimulating 2 months later. The best effects were achieved in 2 months after brassinolide stimulating. The conclusions prove that morphological characters and the main chemical constituents accumulation of G. uralensis could be effected by exogenous BR stimulation in certain case.

BACKGROUNDDespite extensive research, the mechanism of immature dendritic cells (DCs) induced immune hyporesponsiveness remains incompletely understood.

METHODSRecipient DCs from C3H mouse bone marrow cells were incubated with donor antigen from splenic lymphocytes of C57BL/6 mouse; these DCs were transfected with CD80/86 specific siRNA using lentiviral vectors. Flow cytometry was used to evaluate expression of CD80/86 on the antigen-pulsed recipient DCs. Immune regulatory activity was examined by mixed lymphocyte reaction, in which irradiated DCs were cultured with C3H spleen T cells. After the reaction, interleukin (IL)-2, IL-4, IL-10, and interferon (INF)-γ levels of mixed lymphocyte reaction culture supernatant were measured by enzyme-linked immunosorbent assay. The apoptotic T lymphocytes were identified by Annexin V and CD3 staining.

RESULTSThere was a significant inhibition of CD80/86 expression in DCs transfected with CD80/86 lentiviral vectors compared with the control groups (P < 0.05), indicating the specificity of RNA interference. Enzyme-linked immunosorbent assay results showed a significant reduction of INF-γ, IL-2 and IL-10 in the CD80/86 lentivirus transfected group compared to the control groups (P < 0.05). There was no significant difference in IL-4 levels between the groups (P > 0.05). We also showed that CD80/86 low DCs loaded with alloantigen (1) stimulated low T cell proliferative responses via the indirect recognition pathway and (2) enhanced apoptotic activity (P < 0.05) in co-cultured T cells.

CONCLUSIONSLentiviral vector transfection can effectively and specifically knock down target genes in DCs. The CD80/86 low DCs may show tolerogenic activity via induction of T-cell apoptosis, thereby modulating the activity of recipient-derived DCs. The use of this approach may potentially be clinically applicable.

Animals ; Apoptosis ; B7-1 Antigen ; genetics ; physiology ; B7-2 Antigen ; genetics ; physiology ; Dendritic Cells ; immunology ; Lentivirus ; genetics ; Lymphocyte Activation ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; RNA Interference ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo study the antifertility and anti-inflammatory effects of compound Kucen gel in vivo.

METHODSAs antifertility experiment, we randomly divided 60 female SD rats into six groups of equal number: normal saline, blank gel, low-, medium- and high-dose compound Kucen gel (0.05, 0.10 and 0.15 g/g), and positive control (4% nonoxynol gel) to receive intravaginal administration of 200 microl of respective agent, followed by copulation with male rats in a 1:1 ratio. At 12 days after successful mating, the female rats were dissected for calculation of the embryos and the rate of contraception. As an anti-inflammatory trial, we established a mouse model of inflammation by applying xylene to the pinna, and equally randomized 60 Kunming mice to six groups as in the former experiment. We determined the degrees and average rates of swelling inhibition in the left ear.

RESULTSHigh-dose compound Kucen gel achieved a fertility-inhibition rate of 100% in the female rats, the number of embryos significantly lower than in the normal saline group (0.00 +/- 0.00 vs 11.00 +/- 2. 00, P < 0.05), but with no statistically insignificant difference from that of the positive control (0.00 +/- 0.00, P > 0.05). High-dose compound Kucen gel also markedly suppressed swelling in the left ear of the mice, with an inhibition rate of 52.3%, the average swelling degree significantly lower than in the normal saline group (10.17 +/- 2.56 vs 21.32 +/- 3.17, P < 0.01), but not remarkably different from that of the positive control (8.53 +/- 1.89, P > 0.05).

CONCLUSIONCompound Kucen gel, with its strong antifertility and anti-inflammatory effects, deserves further study and clinical application.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Contraceptive Agents ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gels ; Inflammation ; Male ; Mice ; Mice, Inbred Strains ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the spermicidal effect of alcohol extracts from different ratios of Sophora flavescens Ait/Chinese Bulbul in vitro.

METHODSSemen samples aseptically obtained by masturbation and prepared by density gradient centrifugation from 15 healthy men were incubated in the alcohol extracts from 9 different ratios of Sophora flavescens Ait/Chinese Bulbul for 20 seconds, 2 minutes and 4 minutes. Then the motility and movement parameters of the sperm were detected by computer-assisted semen analysis, and the minimal effective concentrations of the instant spermicidal effect of the extracts were determined.

RESULTSAt the ratio of 3:1, the extract at 0.5 mg/ml significantly inhibited the sperm motility and other sperm movement parameters VCL, VSL, VAP, ALH, WOB and MAD, as compared with the control group. The minimal effective concentration of the instant spermicidal effect of the extracts was 3.5 mg/ml at 3:1.

CONCLUSIONThe alcohol extracts from Sophora flavescens Ait and Chinese Bulbul at the ratio of 3:1 have the best spermicidal effect in vitro.

Adult ; Humans ; Male ; Plant Extracts ; pharmacology ; Pulsatilla ; Semen Analysis ; Sophora ; Sperm Motility ; drug effects ; Spermatocidal Agents ; pharmacology ; Spermatozoa ; drug effects ; Young Adult

OBJECTIVETo investigate the early changes of some immunological function of T-cell in chromate workers.

METHODSA total of 115 workers exposed to different levels of soluble chromate were enrolled in exposed group; while 90 non-exposure workers who lived far away from the chromate plant were enrolled as control. The air concentration of soluble chromate was determined by atomic absorption spectrometry. CD3(+), CD3(+)CD4(+), CD3(+)CD8(+) and CD4(+)/CD8(+) of T-cell were determined by flow cytometry analysis.

RESULTSThe individual air chromate concentration in the exposed group was (27.51 +/- 33.25) microg/m(3), and the control group was (0.16 +/- 0.15) microg/m(3). The significant difference between the two groups was observed (z = 8.045, P < 0.01). The levels of the lymphocyte subsets (CD3(+), CD3(+)CD4(+), CD3(+)CD8(+) and CD4(+)/CD8(+)) in exposed group were (30.08 +/- 17.75)%, (1.04 +/- 1.73)%, (11.94 +/- 9.78)%, 0.10 +/- 0.14. While, those of control group were (63.00 +/- 13.57)%, (30.51 +/- 5.16)%, (14.82 +/- 4.59)%, 2.17 +/- 0.53, higher than that of the exposed group (z values were 4.484, 5.227, 1.976, -5.218, respectively, P < 0.05).

CONCLUSIONOn the basis of individual air monitoring, the cellular immune function affected by soluble chromate is mainly based on T lymphocyte inhibition. The indicators CD3(+)CD4(+) mentioned above may be considered as efficient biomarkers in further research.

Adult ; CD4-CD8 Ratio ; Case-Control Studies ; Chromates ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; T-Cell Antigen Receptor Specificity ; drug effects ; immunology ; T-Lymphocytes ; drug effects ; immunology

OBJECTIVETo investigate the expression of gene TRAIL driven by human telomerase reverse transcriptase (hTERT) promoter in SACC-83 cell tumor necrosis factor related apoptosis in dncing ligand.

METHODSAfter pACTERT-TRAIL plasmid transfected was into SACC-83 and HEL cells through liposome, the expression of TRAIL was examined using RT-PCR technique, the cells' survival rate by methyl thiazolyl tetrazolium (MTT) method and apoptosis rate by flow cytometry.

RESULTSExpression of extrinsic TRAIL gene driven by hTERT promoter was detected in SACC-83 cells, and not detected in human embryonic lung fibroblast (HEL) cells. After transfection of pACTERT-TRAIL, the proliferation of SACC-83 cells was significantly inhibited, and its apoptotic rate was promoted, whereas no inhibited effect was observed on HEL cells and its apoptotic rate showed little change.

CONCLUSIONShTERT promoter can be used to induce tumor-specific expression of TRAIL gene and apoptosis in SACC-83 cells.

Apoptosis ; Carcinoma, Adenoid Cystic ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Gene Targeting ; Genetic Vectors ; Humans ; Salivary Gland Neoplasms ; genetics ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; Telomerase ; genetics ; Transfection

OBJECTIVETo study the apoptotic effect on the squamous cell carcinoma cell line TCa83 induced by recombined adenovirus vector containing TRAIL gene and CMV promoter.

METHODSThe TCa83 cell line was firstly infected with different titre of AdCMV-EGFP containing enhanced green fluorescence protein gene (EGFP) as control, and investigated the transducing rate through fluorescence to obtain the definite titre. Then TCa83 cell line was infected with AdCMV-TRAIL in proper titre, and TRAIL gene was detected by means of RT-PCR. After TCa83 cell line was infected with AdCMV-TRAIL and AdCMV-EGFP at day 1, 3, 5, 7, the activity of TCa83 cell line were evaluated by MIT and the apoptosis were detected by flow cytometer.

RESULTSProper titre was of 1,000 particles/cell, and TCa83 cell line could be infected 100% in this titre. TRAIL gene was detected by RT-PCR after infected with AdCMV-TRAIL. The activity of TCa83 decreased in both groups, but the AdCMV-TRAIL group decreased more sharply than AdCMV-EGFP group (P < 0.001). Both AdCMV-TRAIL and AdCMV-EGFP could lead to apoptosis of TCa83 cells, but the AdCMV-TRAIL, function stronger than AdCMV-EGFP. Especially there was remarkable statistic difference between two groups (P < 0.0001).

CONCLUSIONAdCMV-TRAIL could effectively decrease the activity of TCa83 cell line and induce apoptosis.

Adenoviridae ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Line ; Genetic Vectors ; Green Fluorescent Proteins ; Humans ; Promoter Regions, Genetic