Objective To investigate the sensitivity of 5 kinds of chemotherapeutic drugs on human colorectal cancer in vivo. Methods Xenografts in nude mice were set up by tumor tissues from 9 patients with colorectal cancer and nude mice were divided into 6 groups randomly， receiving saline （control group）， 5-fluorouracil （5-FU group）， doxorubicin（ADM group）， mitomycin （MMC group）， oxaliplatin （LOHP group）， and irinotecan （CPT-11 group）， respectively. The inhibitive rates （IR） of xenografts in 5 groups for each patient were calculated. Results The lowest and highest IR of 5 groups were 23.6% and 54.9% in 5-FU group， 23.7% and 69.5% in LOPH group， 23.6% and 82.6% in CPT-11group， 24.1% and 48.1% in MMC group， 5.8% and 20.7% in ADM group， respectively. The IR exceeded 40.0% in 7 patients of LOHP group， 6 patients of CPT-11 group， 4 patients of 5-FU group， and 1 patient of MMC group， respec-tively. Of 9 patients， the IR exceeded 40.0% to 3 kinds of drugs in 3 patients， to 2 kinds of drugs in 4 patients， the IR didn’t exceed 30.0% to 4 kinds of drug （IR was 82.6% to CPT-11） in 1 patient， and the IR didn’t exceed 31.0% to all 5 kinds of drugs in 1 patient. There were statistical differences on the IR of 5 kinds of drugs （H=24.061 2， P=0.000 1）. IR of ADM group was statistical lower than 5-FU group， MMC group， LOHP group， and CPT-11 group （P＜0.05），but there were no statistical differences between 5-FU group， MMC group， LOHP group， and CPT-11 group （P＞0.05）. Conclusions The xenografts from same patient have different sensitivity to different chemotherapy drugs， and the same chemotherapy drug corresponds to different IR in different patients. The IR of LOHP and CPT-11 are the highest， following by 5-FU and MMC.
Near-infrared fluorescence imaging technology, which possesses superior advantages including real-time and fast imaging, high spatial and temporal resolution, and deep tissue penetration, shows great potential for tumor imaging in vivo and therapy. Ⅰ-Ⅲ-Ⅵ quantum dots exhibit high brightness, broad excitation, easily tunable emission wavelength and superior stability, and do not contain highly toxic heavy metal elements such as cadmium or lead. These advantages make Ⅰ-Ⅲ-Ⅵ quantum dots attract widespread attention in biomedical field. This review summarizes the recent advances in the controlled synthesis of Ⅰ-Ⅲ-Ⅵ quantum dots and their applications in tumor imaging in vivo and therapy. Firstly, the organic-phase and aqueous-phase synthesis of Ⅰ-Ⅲ-Ⅵ quantum dots as well as the strategies for regulating the near-infrared photoluminescence are briefly introduced; secondly, representative biomedical applications of near-infrared-emitting cadmium-free quantum dots including early diagnosis of tumor, lymphatic imaging, drug delivery, photothermal and photodynamic therapy are emphatically discussed; lastly, perspectives on the future directions of developing quantum dots for biomedical application and the faced challenges are discussed. This paper may provide guidance and reference for further research and clinical translation of cadmium-free quantum dots in tumor diagnosis and treatment.
ObjectiveTo evaluate the most efficient method for transfection of human umbilical cord mesenchymal stem cells (HUMCSs) in vivo. MethodsHUCMSCs were isolated from human umbilical cord and cultured, which were labelled by PKH26 and lentivirus-GFP, then were observed by using a fluorescence microscope. Sixty SD rats were randomly divided into PKH26 transfection group and lentivirus-GFP transfection group. The right hepatic lobe of rat was resected, then the transfected stem cells were injected into portal vein. The rats were sacrificed on day 3, 8, and 13 after transfection. The liver specimens were observed by using a fluorescence microscope. Flow cytometry was used to evaluate the percentage of transfected stem cells and the apoptotic stem cells. ResultsThe third generation of HUCMSCs labelled by PKH26 and lentivirus-GFP were spindle shaped. PKH26 red dye was evenly distributed in the cell membrane of HUCMSCs and could be clearly labelled. The HUCMSCs labelled by lentivirus-GFP were green fluorescence under the fluorescence microscope, and it was clear and stable. The HUCMSCs were clear and could be clearly distinguished on day 3 after transfection by two methods in vivo. As the time went by, red was faded and blurred, then was gradually disappeared on day 13 after transfection in the HUCMSCs stansfected by PKH26; but the color in the HUCMSCs stansfected by lentivirus-GFP were clear at all the time points. The transfection rate of the lentivirus-GFP was significantly higher that that of the PKH26 (P < 0.05), the rate of apoptotic stem cells had no significant differences at all the time points between these two groups (P > 0.05). ConclusionLentivirus-GFP transfection is a higher efficient method for stem cell labelling in vivo, it could be used to observe transplantation cells for a long time in future.
ObjectiveThis study construct a pulmonary fibrosis model in vivo to study anti-pulmonary fibrosis effect of ampelopsis.MethodsWe constructed a pulmonary fibrosis model by bleomycin in BALB/c mice. The mice were divided by weight random number table into a blank control group, a model control group, a dexamethasone treatment group (intervened with dexamethasone in a dose of 2.5 mg/kg), and three ampelopsis treatment groups intervened with ampelopsis in dose of 200, 100, and 50 mg/kg, respectively. Bleomycin solution (3 mg/kg) was intratracheally injected respectively on 1st and 14th day, except the blank group. Twenty-eight days later, the relevant indicators were collected, including respiratory function (airway resistance, dynamic lung compliance, maximal ventilator volume), level of hydroxyproline and histopathological changes in the lungs.ResultsAfter 28 days, the model control group mice had severe respiratory resistance, dynamic lung compliance and maximal ventilator volume were decreased. The high dose ampelopsis treatment could enhance respiratory function (P<0.05). Lung coefficient was lower in the treatment groups than that in the model control group (P<0.05). The hydroxyproline of the treatment groups was less than that of the model control group (P<0.05). Histopathological examination showed that the degree of fibrosis increased in the model control group (P<0.05), but decreased in the treatment groups (P<0.05).ConclusionAmpelopsis can resist bleomycin-induced pulmonary fibrosis in mice, relieve the symptoms of respiratory failure, reduce the formation of collagen, and produce anti-pulmonary fibrosis effect.
ObjectiveTo design a method for observing pulmonary microcirculation in vivo in a native tissue environment, due to the high incidence of lung diseases and the advantages of animal experiments in vivo.MethodsTen BALB/c male mice were randomly divided into group A and group B, with five mice in each group. A self-made apparatus was used to control the movement towards local lung tissues in order to get a stabilized observation plane, and then a 5-minute video was shot with laser confocal scanning microscope. During the filming, the mice in group A were injected with fluorescein isothiocyanate-dextran via the tail vein, and the mice in group B were injected with green fluorescent protein-platelets (extracted from the blood of tie2-cre&rosa26-tomato-EGFP transgenic black C57 male mice). The data of group A was used to observe the lungs perfusion and the damage to tissue by this method, and the data of group B was used to observe the movement of platelets.ResultsImage of lung structure obtained by this method was clear and stable. Mean areas of alveolus in an instant and at the 30th, 60th, 120th, 180th, and 300th second were (1 603±181), (1 588±183), (1 528±363), (1 506±353), (1 437±369), (1 549±307) μm2, respectively, and there were no significant differences between each time point (P>0.05). The video was smooth, the rapid movement of platelets was recorded and the particles were clear and without tailing; after the observation, hematoxylin-eosin staining showed no obvious damage to the lung tissue.ConclusionThe method can be used for the observation and research of the lung microcirculatory system in the living state of the mouse, and provides a methodological basis for studies of other lung diseases in vivo.
Objective To review the in vivo imaging research progress of two-photon microscopy (TPM) in spinal cord. Methods The recent literature concerning in vivo two-photon imaging of axon, microglia, and calcium in transgenic mice spinal cord was extensively consulted and reviewed. Results In vivo two-photon imaging of spinal cord provide dynamic information about axonal degeneration and regeneration, microglial accumulation, and calcium influx after spinal cord injury. Conclusion TPM in vivo imaging study on spinal cord will provide theoretical foundation for pathophysiologic process of spinal cord injury.
Objective To investigate the in vivo degradable properties of new calcium phosphate cement (CPC) containing poly lactic-co-glycolic acid (PLGA) so as to lay a foundation for the future clinical application. Methods A novel CPC containing PLGA (CPC/PLGA) was prepared according to a ratio of 45% dicalcium phosphate anhydrous ∶ 45% partially crystallized calcium phosphates ∶ 10% PLGA. Thirty-two adult New Zealand rabbits (weighing 2.2-3.0 kg, male or female in half) were divided into the experimental group (n=17) and the control group (n=15). The bone defect models of the bilateral femoral condyles (4.5 mm in diameter and 1.5 cm in depth) were made by drilling hole. Defect at the right side was repaired with CPC/ PLGA in the experimental group and with CPC in the control group, while defect at the left side was not treated as blank control. The general condition of rabbits was observed after operation; the histological observation and bone histomorphometric analysis were performed at 2, 4, 8, 16, and 24 weeks; and scanning electronic microscope (SEM) observation was performed at 8 and 16 weeks after operation. Results All rabbits survived to the end of experiment. The histological observation showed: CPC/PLGA degraded gradually, and the new-born bone trabecula ingrew; bone trabeculae became rough and b; and CPC/PLGA almost biodegraded at 24 weeks in the experimental group. The CPC degradation was much slower in the control group than in the experimental group. The total bone tissue percentage was 44.9% ± 23.7% in the experimental group, and 25.7% ± 10.9% in the control group, showing significant difference between 2 groups (t=3.302, P=0.001); and the bone tissue percentage showed significant difference between 2 groups at 8, 16, and 24 weeks (P lt; 0.05). The results of SEM observation showed that the pore size was 100-300 μm at 8 weeks after operation, new-born bone trabecula grew into the pores and combined bly with residual cement in the experimental group. Conclusion Novel CPC/PLGA has good in vivo degradable properties, and it can be an ideal bone substitute in future clinical application.
ObjectiveTo explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. MethodsBreast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. ResultsExcept for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. ConclusionsCAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.
ObjectiveTo review the application and research progress of in vivo bioreactor as vascularization strategies in bone tissue engineering. MethodsThe original articles about in vivo bioreactor that can enhance vascularization of tissue engineered bone were extensively reviewed and analyzed. ResultsThe in vivo bioreactor can be created by periosteum, muscle, muscularis membrane, and fascia flap as well as biomaterials. Using in vivo bioreactor can effectively promote the establishment of a microcirculation in the tissue engineered bones, especially for large bone defects. However, main correlative researches, currently, are focused on animal experiments, more clinical trials will be carried out in the future. ConclusionWith the rapid development of related technologies of bone tissue engineering, the use of in vivo bioreactor will to a large extent solve the bottleneck limitations and has the potential values for clinical application.
The human femur is in a relatively complex mechanical environment, subject to the combined effects of multiple factors such as mechanical loads from movement and weight-bearing, as well as changes in the body fluid environment in daily life. In in vitro testing cases of the femur (e.g., testing of distal femoral fractures), changes in load conditions usually significantly affect the mechanical properties of the overall structure. However, there is currently no systematic evaluation standard for in vitro mechanical performance testing of the femur. Therefore, this paper established four human femur models (model A~model D) constructed based on computed tomography (CT) under different load environments, as well as two artificially synthesized femur models (the finite-element model and the experimental model) under the same load environment. Among them, for the human femur models, model A was configured to apply hip joint contact forces together with all muscle forces to approximate the real in vivo mechanical environment, model B was applied with hip joint contact force and abductor muscle force, model C was only applied with hip joint contact force, and model D was subjected to an equivalent resultant force. For the artificially synthesized femur models, both the finite-element model and the experimental model were applied with the same equivalent resultant force as model D. Comparative analyses revealed that model D exhibited femoral head displacement and stress-strain distributions similar to Model A, indicating its suitability as an equivalent in vitro test model. Further comparison between the finite-element and experimental synthetic femur models yielded consistent mechanical responses, thereby validating the equivalent model. In summary, it is hoped that the findings of this study will provide a reference for establishing a systematic, tiered preclinical evaluation system for hip prostheses/implants in the future.