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.
Objective To research in vitro biocompatibility of silicon containing micro-arc oxidation (MAO) coated magnesium alloy ZK60 with osteoblasts. Methods The surface microstructure of silicon containing MAO coated magnesium alloy ZK60 was observed by a scanning electron microscopy (SEM), and chemical composition of the coating surface was determined by energy dispersive spectrum analysis. The experiments were divided into 4 groups: silicon containing MAO coated magnesium alloy ZK60 group (group A), uncoated magnesium alloy ZK60 group (group B), titanium alloy group (group C), and negative control group (group D). Extracts were prepared respectively with the surface area to extraction medium ratio (1.25 cm2/ mL) according to ISO 10993-12 standard in groups A, B, and C, and were used to culture osteoblasts MC3T3-E1. The α-MEM medium supplemented with 10% fetal bovine serum was used as negative control in group D. The cell morphology was observed by inverted phase contrast microscopy. MTT assay was used to determine the cell viability. The activity of alkaline phosphatase (ALP) was detected. Cell attachment morphology on the surface of different samples was observed by SEM. The capability of protein adsorption of the coating surface was assayed, then DAPI and calcein-AM/ethidium homodimer 1 (calcein-AM/EthD-1) staining were carried out to observe cell adhesion and growth status. Results The surface characterization showed a rough and porous layer with major composition of Mg, O, and Si on the surface of silicon containing MAO coated magnesium alloy ZK60 by SEM. After cultured with the extract, cells grew well and presented good shape in all groups by inverted phase contrast microscopy, group A was even better than the other groups. At 5 days, MTT assay showed that group A presented a higher cell proliferation than the other groups (P lt; 0.05). Osteoblasts in groups A and C presented a better cell extension than group B under SEM, and group A exhibited better cell adhesion and affinity. Protein adsorption in group A [ (152.7 ± 6.3) µg/mL] was significantly higher than that of group B [(96.3 ± 3.9) µg/mL] and group C [ (96.1 ± 8.7) µg/mL] (P lt; 0.05). At each time point, the adherent cells on the sample surface of group A were significantly more than those of groups B and C (P lt; 0.05). The calcein-AM/EthD-1 staining showed that groups A and C presented better cell adhesion and growth status than group B. The ALP activities in groups A and B were 15.55 ± 0.29 and 13.75 ± 0.44 respectively, which were significantly higher than those in group C (10.43 ± 0.79) and group D (10.73 ± 0.47) (P lt; 0.05), and group A was significantly higher than group B (P lt; 0.05). Conclusion The silicon containing MAO coated magnesium alloy ZK60 has obvious promoting effects on the proliferation, adhesion, and differentiation of osteoblasts, showing a good biocompatibility, so it might be an ideal surface modification method on magnesium alloys.
Cell migration is defined as the directional movement of cells toward a specific chemical concentration gradient, which plays a crucial role in embryo development, wound healing and tumor metastasis. However, current research methods showed low flux and are only suitable for single-factor assessment, and it was difficult to comprehensively consider the effects of other parameters such as different concentration gradients on cell migration behavior. In this paper, a four-channel microfluidic chip was designed. Its characteristics were as follows: it relied on laminar flow and diffusion mechanisms to establish and maintain a concentration gradient; it was suitable for observation of cell migration in different concentration gradient environment under a single microscope field; four cell isolation zones (20 μm width) were integrated into the microfluidic device to calibrate the initial cell position, which ensured the accuracy of the experimental results. In particular, we used COMSOL Multiphysics software to simulate the structure of the chip, which demonstrated the necessity of designing S-shaped microchannel and horizontal pressure balance channel to maintain concentration gradient. Finally, neutrophils were incubated with advanced glycation end products (AGEs, 0, 0.2, 0.5, 1.0 μmol·L−1), which were closely related to diabetes mellitus and its complications. The migration behavior of incubated neutrophils was studied in the 100 nmol·L−1 of chemokine (N-formylmethionyl-leucyl-phenyl-alanine) concentration gradient. The results prove the reliability and practicability of the microfluidic chip.
Reduced chemotactic migration of polymorphonuclear neutrophil (PMN) in sepsis patients leads to decreased bacterial clearance and accelerates the progression of sepsis disease. Quantification of PMN chemotaxis in sepsis patients can help characterize the immune health of sepsis patients. Microfluidic microarrays have been widely used for cell chemotaxis analysis because of the advantages of low reagent consumption, near-physiological environment, and visualization of the migration process. Currently, the study of PMN chemotaxis using microfluidic chips is mainly limited by the cumbersome cell separation operation and low throughput of microfluidic chips. In this paper, we first designed an inertial cell sorting chip to achieve label-free separation of the two major cell types by using the basic principle that leukocytes (mainly granulocytes, lymphocytes and monocytes) and erythrocytes move to different positions of the spiral microchannel when they move in the spiral microchannel under different strength of inertial force and Dean's resistance. Subsequently, in this paper, we designed a multi-channel cell migration chip and constructed a microfluidic PMN inertial label-free sorting and chemotaxis analysis platform. The inertial cell sorting chip separates leukocyte populations and then injects them into the multi-channel cell migration chip, which can complete the chemotaxis test of PMN to chemotactic peptide (fMLP) within 15 min. The remaining cells, such as monocytes with slow motility and lymphocytes that require pre-activation with proliferative culture, do not undergo significant chemotactic migration. The test results of sepsis patients (n=6) and healthy volunteers (n=3) recruited in this study showed that the chemotaxis index (CI) and migration velocity (v) of PMN from sepsis patients were significantly weaker than those from healthy volunteers. In conclusion, the microfluidic PMN inertial label-free sorting and chemotaxis analysis platform constructed in this paper can be used as a new tool for cell label-free sorting and migration studies.
ObjectiveTo investigate the anesthesia management of transcatheter ultrasound-guided percutaneous interventional therapy for pediatric patients with congenital heart disease at a mobile surgical platform. Methods From March to July 2023, 13 patients in remote areas underwent interventional surgery on the mobile truck operating platform. The patients undergoing general anesthesia using non-tracheal intubation were collected. ResultsFinally, 8 patients received monitored anesthesia care (MAC) with local anesthesia-assisted sedation and analgesia drugs under the supervision of anesthesiologists (general anesthesia using non-tracheal intubation), due to the patients having difficulty cooperating with the surgery (young age, nervous mood, and crying), including 5 males and 3 females with an average age of 6.95±3.29 years and an average weight of 19.50±6.04 kg. There were 6 patients diagnosed with atrial septal defect, 1 patient with residual shunt after patent ductus arteriosus ligation, and 1 patient with severe pulmonary stenosis by transthoracic ultrasonography. The surgical process was smooth, analgesia was perfect, anesthesia and surgical effect were satisfactory, postoperative recovery was satisfactory, and there were no surgical or anesthesia complications. The anesthesia time was 41.53±8.62 min, the operation time was 39.88±8.52 min, and the recovery time was 41.50±14.56 min. Conclusion Transthoracic ultrasound-guided interventional surgery is a minimally invasive approach for congenital heart disease, offering the advantages of zero radiation exposure. Non-tracheal general anesthesia preserved spontaneous breathing can be safely and effectively administered to pediatric patients who cannot cooperate in mobile operating platform.