Pancreatic cancer is a kind of highly malignant tumor with a low survival rate and poor prognosis. The effectiveness of gemcitabine as a first-line chemotherapy drug is limited; however, it can activate dendritic cells and improve antigen presentation which increase the sensitivity of tumor cell to immunotherapy. Although immunotherapy has made some advancements in cancer treatment, the therapeutic benefit of programmed cell death receptor 1/programmed death receptor-ligand 1 (PD-1/PD-L1) blockade therapy remains relatively low. The chemokine C-X-C chemokine ligand 12 (CXCL12) contributes to an immunosuppressive tumor microenvironment by recruiting immunosuppressive cells. The receptor C-X-C motif chemokine receptor 4 (CXCR4), highly expressed in various tumors including pancreatic cancer, plays a crucial role in tumor development and progression. In this study, the anti-tumor immune response of human peripheral blood mononuclear cell (hPBMC) was enhanced using the combination of BsNb PX4 (anti-PD-L1&CXCR4 bispecific nanobody) and gemcitabine. In a co-culture system of gemcitabine-pretreated hPBMCs with tumor cells, the BsNb PX4 synergized gemcitabine to improve the cytotoxic activity of hPBMCs against tumor cells. Flow cytometry analysis confirmed increased ratio of CD8⁺ to CD4⁺ T cells in combination treatment. In NOD/SCID mice bearing pancreatic cancer, the combination treatment exhibited more infiltration of CD8⁺ T cells into tumor tissues, contributing to an effective anti-tumor response. This study presents potential new therapies for the treatment of pancreatic cancer. Ethical approval was obtained for collection of hPBMC samples from the Local Ethics Committee of Shanghai Jiao Tong University. All animal experiments were approved by the Animal Ethic Committee of Shanghai Jiao Tong University (authorizing number: A2024246).

Fifty-six patients with Graves' ophthalmopathy(GO)were treated with antithyroid drug and oral prednisone for three months,TSH receptor antibody(TRAb)level was reduced,GO activity and severity of some patients were ameliorated but still positively associated with TRAb.It suggests that TRAb not only triggers off GO but also plays a possible role in the maintenance of the autoimmune process in GO.

In this study, wheat germ agglutinin (WGA), tomato lectin (TL) and asparagus pea lectin (AL) were covalently coupled to conventional poly lactic-co-glycolic acid (PLGA) nanoparticles using a carbodiimide method to take the bioadhesive properties. The influences of the amounts of activating agents and lectins, as well as the activating time and incubating time on the effect of lectin conjugating were investigated to optimize the preparation conditions. The mean diameters of the performed nanoparticles with or without lectin conjugation ranged from (140.7 +/- 5.7) nm to (245.6 +/- 18.3) nm. The yields of lectin conjugating and the lectin surface concentrations on nanoparticles were determined by Lowry's methods, and were calculated to be (18.97 +/- 2.9)% - (20.15 +/- 2.4)% and (9.46 +/- 1.45)--(10.05 +/- 1.19) microg x mg(-1), respectively. The in vitro bioadhesive activities of nanoparticles were evaluated by pig gastric mucin (PM) binding experiments. After incubation at room temperature for 60 min, the equilibria of binding between nanoparticles and PM reached. The percentages of the bulk PM which had interacted with different lectin-conjugated PLGA nanoparticles were 15.5%, 12.1% and 11.8%, respectively. The conjugation of lectin enhanced the interaction about 2.4 - 3.2 fold compared with that of the non-conjugated one. A mathematical model was used based on the Langmuir equation, and the rate constants of interaction (k) were calculated to be 2.373 x 10(-3), 1.536 x 10(-3) and 1.714 x 10(-3) (microg x min/mL)(-1), respectively. These interactions could be competitively inhibited by their corresponding sugars of lectins. The results suggested that lectin-conjugated PLGA nanoparticles greatly promoted the interaction with PM in vitro compared with the conventional PLGA nanoparticles, thus would improve the bioadhesion on gastrointestinal mucosa after oral administration resulting in a prolonged residence time in the gastrointestinal tract.
Adhesiveness ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Gastric Mucins ; metabolism ; Lactic Acid ; chemistry ; metabolism ; Nanoparticles ; Plant Lectins ; chemistry ; metabolism ; Polyglycolic Acid ; chemistry ; metabolism ; Protein Binding ; Wheat Germ Agglutinins ; chemistry ; metabolism

AIMUnderstanding the response of mesenchymal stem cells (MSCs) to mechanical strain and their consequent gene expression patterns will broaden our knowledge of the mechanobiology of distraction osteogenesis.

METHODOLOGYIn this study, a single period of cyclic mechanical stretch (0.5 Hz, 2,000 microepsilon) was performed on rat bone marrow MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity was examined. The mRNA expression of six bone-related genes (Ets-1, bFGF, IGF-II, TGF-beta, Cbfa1 and ALP) was detected using real-time quantitative RT-PCR.

RESULTSThe results showed that mechanical strain can promote MSCs proliferation, increase ALP activity, and up-regulate the expression of these genes. A significant increase in Ets-1 expression was detected immediately after mechanical stimulation, but Cbfa1 expression became elevated later. The temporal expression pattern of ALP coincided perfectly with Cbfa1.

CONCLUSIONThe results of this study suggest that mechanical strain may act as a stimulator to induce differentiation of MSCs into osteoblasts, and that these bone-related genes may play different roles in the response of MSCs to mechanical stimulation.

Alkaline Phosphatase ; analysis ; Animals ; Antigens, Surface ; analysis ; Biomechanical Phenomena ; Bone Marrow Cells ; physiology ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Core Binding Factor Alpha 1 Subunit ; analysis ; Fibroblast Growth Factor 2 ; analysis ; Insulin-Like Growth Factor II ; analysis ; Mesenchymal Stromal Cells ; physiology ; Osteoblasts ; physiology ; Osteogenesis, Distraction ; Pluripotent Stem Cells ; physiology ; Proto-Oncogene Protein c-ets-1 ; analysis ; Rats ; Stress, Mechanical ; Transforming Growth Factor beta ; analysis ; Up-Regulation ; physiology

OBJECTIVETo investigate the resistance to cyclic fatigue of maxillary incisors with flared canals restored with different post-and-core materials.

METHODSThirty human maxillary central incisors were assigned randomly to two main groups (non-ferrule and 1-mm ferrule) of 15. Each main group was then divided into 3 groups of 5 specimens and restored with custom cast post-and-core (MPC), resin composite post-and-core (RCP), and resin composite core combined with prefabricated carbon fiber post (FRC), respectively. Every group was subjected to a cyclic fatigue test, and recorded the numbers of load cycles that occurred tooth fracture. All data were analyzed with two-way ANOVA and the Tukey HSD test at alpha = 0.05.

RESULTSFRC revealed significantly higher fatigue strength than the other groups (P < 0.05). Preparing a dentin ferrule increased significantly fatigue resistance (P < 0.05). The favorable (retrieval) fracture pattern of the tested specimens was discovered only in RCP.

CONCLUSIONSUsing FRC may get a long fatigue life in restoring pulpless teeth with flared canals. Dentin ferrule preparation is necessary to enhance resistance of the restorations to cyclic fatigue.

Carbon ; Chromium Alloys ; Composite Resins ; Dental Materials ; Dental Pulp Cavity ; pathology ; Humans ; Materials Testing ; Post and Core Technique ; Tooth, Nonvital ; therapy

OBJECTIVETo observe the effects of mesenchymal stem cells (MSCs)on cranial suture under mechanical strain in growing goats.

METHODS10 growing goats were used in this study. A customized distractor was used for distraction of the coronal suture at a rate of 0.4 mm/day for 8 days. The experimental group(5 goats) was injected with autologous MSCs into the distracted region, whereas the control group (5 goats) with injection of physiological saline. All animals were killed at 4 weeks after the end of distraction. Scanning electron microscopy and histological analysis were taken to observe the samples.

RESULTS4 weeks after the end of distraction, the cranial sutures in all animals were separated successfully. The new bone formation at the edge of suture in the experimental group was superior to that in the control group.

CONCLUSIONAutologous MSCs transplantation may promote the cranial suture distraction osteogenesis in the growing goats.

Animals ; Cranial Sutures ; Goats ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Osteogenesis, Distraction ; Skull

OBJECTIVETo construct recombinant plasmid pEGFP-BMP7 and determine its expression in rat bone marrow mesenchymal stem cells (MSCs) in vitro.

METHODScDNA of target gene was obtained from neonatal rat kidney by RT-PCR. After sequencing the target gene, the cDNA was subcloned into a eukaryote plasmid pEGFP-N1 by directed cloning and then digested with two restrictive endonucleases to verify the correctiveness of the recombinant plasmid pEGFP-BMP7. Rat bone marrow MSCs were transiently transfected with the pEGFP-BMP7 and transfection efficiency of the Green Fluorescent Protein (GFP) was determined. RT-PCR and immunocytochemical analysis were also performed to detect the expression of BMP7 in rat MSCs.

RESULTS1 311 bp cDNA fragment was obtained by RT-PCR and sequence analysis showed it matched perfectly with that of rat BMP7 gene except a single nucleotide change at 756 bp from T to A. Digestion of the recombinant plasmid showed two 1.3 kb and 4.7 kb fragments and their size were same as those of BMP7 and pEGFP. This indicated that BMP7 cDNA was successfully subcloned into pEGFP. Transient transfection showed an efficiency of 33% at day 2 in rat MSCs. After transfection, transcription of BMP7 was detected in MSCs and expression of BMP7 protein was also verified.

CONCLUSIONRecombinant eukaryote plasmid pEGFP-BMP7 was successfully constructed and expressed in rat bone marrow MSCs. This procedure may provide a unique method for stimulation of callus formation in distraction osteogenesis and reconstruction of craniofacial bone defects.

Animals ; Bone Marrow Cells ; Bone Morphogenetic Protein 7 ; Genetic Vectors ; Green Fluorescent Proteins ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Plasmids ; Rats ; Transfection

Glutamate (Glu) is the major afferent excitatory neurotransmitter in the auditory system, and excessive Glu may play an important role in cochlear dysfunction. It is unclear how excessive Glu plays roles in cochlear dysfunction in cochlear organotypic cultures. In this study neonatal rat cochlear organotypic cultures were prepared, and then the cochlear tissues were incubated with a new medium containing specific concentrations of Glu (0.1, 0.5, 1, 10 or 20 mmol/L) for 24 h, or incubated with the medium containing a concentration of 20 mmol/L Glu for 6, 12, 24 or 72 h, respectively. It was found that when the cochlear tissues were cultured for 24 h, the inner hair cells (IHCs) were damaged at the concentration of 0.5 mmol/L Glu, and with the increases of the concentrations, the injury was gradually aggravated, and 20 mmol/L Glu resulted in the significant loss of IHCs. In the 20 mmol/L Glu groups, the stereocilia bundles were missing or disarrayed on a few IHCs after culture for 6 h and the damage effect was time-dependent. The missing of IHCs was more significant in the basal turn of the cochlea than in the middle turn of the cochlea under the same concentration of Glu exposure. These results suggest that excessive exogenous Glu affects the morphology of IHCs, but not affects the outer hair cells (OHCs) in cochlear organotypic cultures, and the excitotoxic effects are different on IHCs of different parts of the cochlea under the same concentration of Glu exposure.
Animals ; Cochlea ; drug effects ; Dose-Response Relationship, Drug ; Glutamic Acid ; toxicity ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Most studies suggest tooth whitening can create a coarse enamel surface which is likely to attract exogenous dyeing material and form the phenomenon of color relapse. After dental bleaching procedure, the use of resin surface treatment agent is expected to make the enamel surface smooth and alleviate tooth color relapse. OBJECTIVE: To unravel the efficacy of Adper Single Bond 2 Adhesive on color relapse phenomenon after dental bleaching treatment by cold light (xenon laser) whitening.METHODS: Twenty-four extracted teeth were collected and coated with 35% hydrogen peroxide followed by xenon laser whitening. The test group was made up of 12 samples coated with light-cured resin surface treatment agent and the left samples as control group were treated with no-coating agent after tooth whitening. Then the two groups were divided into two subgroups which were soaked in distilled water and tea for aging test respectively. The color differences (ΔE) which provided comparative values for statistical analysis was recorded at 1, 3, 5, 7, 14, 21, 28, 35 and 42 days after the aging test. The enamel surface microstructure of the samples was observed before and after bleaching, after resin agent coating, and 28 days after the aging test.RESULTS AND CONCLUSION: (1) Results from color differences observation: there was no significant difference between samples soaked in distilled water before and after whitening treatment (P > 0.05). Similarly, there was no statistical difference between test group and control group soaked in distilled water (P > 0.05). However, after soaking in tea, the color differences in the test group at 1-42 days showed statistically significant differences from those in the control group (P < 0.05). (2) Results from scanning electron microscope observation: after the tooth whitening producers, the enamel surface was damaged in the presence of cavities. After coated with resin surface treatment agent, the enamel surface became smooth and had few cavities. After soaking in tea, gradient smooth surface, some crack, inconspicuous flake dyeing color layer decomposition could be seen in the test group while rough surface with big holes and mottled dyeing layer were clearly visible in the control group. All samples soaked in distilled water had only a small amount of block dyeing layer, holes, weaker roughness than those soaked in tea. To conclude, Adper Single Bond 2 Adhesive light-cured resin surface treatment agent could weaken color relapse phenomenon after tooth whitening, achieving a smooth enamel surface and reducing dyeing material adhesion.

Objective To investigate effects of angelica polysaccharide on learning and memory abilities, Ach, ChAT, AChE, SOD, MDA in serum, APP and Aβ1-42 in hippocampus in model rats with Alzheimer disease (AD); To explore the mechanism of angelica polysaccharide for the treatment of AD. Methods Seventy SPF Wistar rats were selected for learning and memory ability by water maze. 10 rats were randomly selected (half female and half male) as sham-operation group, and the others were injected with Aβ25-35 by stereotatic techniques, copying AD model rats. 50 rats for learning and memory ability by water maze were successfully divided into model group, positive group, angelica polysaccharide low-, medium-, and high-dose groups, with 10 rats in each group. Rats in model group and sham-operation group were given normal saline for gavage, while rats in medication groups were given relevant medicine for gavage, 2 mL/(100 g?d), for 28 d. The learning and memory ability of rats in each group was tested by Morris water maze during 25-28 days, and the contents of Ach, ChAT, AChE, SOD, MDA in serum and APP and Aβ1-42 in hippocampus were determined. Results Compared with the sham-operation group, the escape latent period of model group was significantly prolonged in place navigation experiment; the target quadrant time was shortened; the latent time for the first time to reach the original escape platform was longer in spatial probe test; the residence time of crossing the original platform position and the target quadrant was shorter; the levels of Ach, the activity of ChAT and SOD in serum decreased; the levels of MDA, the activity of AChE in serum increased; the levels of APP and Aβ1-42 in hippocampus increased, with statistical significance (P<0.05, P<0.01). Compared with model group, the escape latent period of each medication group was shortened in different degrees after the intervention treatment; the residence time of target quadrant was prolonged; the latent time for the first time to reach the original escape platform was shortened; the number of cross platform increased; the levels of Ach, the activity of ChAT and SOD in serum increased; the levels of MDA and the activity of AChE in serum decreased; the levels of APP and Aβ1-42 in hippocampus significantly decreased, with statistical significance (P<0.05, P<0.01). Conclusion Angelica polysaccharide may effectively improve the learning and memory of ability of AD model rats to improve anti-free radical oxidation and promote Aβ metabolism and promote learning and memory ability of AD model rats, which have some preventive and therapeutic effects on AD.