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Objective:To investigate an accurate and quantitative method to measure the eyeball morphological parameters of guinea pigs through a method that combines programmed digital techniques and mathematical geometric principles.Methods:Twenty-two three-week-old clean-grade male tricolor guinea pigs were selected and sacrificed by anesthesia overdose.Eyeballs were enucleated.The horizontal and sagittal images of the eyeball were taken with the high-speed photographic model of 13 million pixels macro meter, and the pictures were imported into pycharm programming software.Using the pre-written analysis program of Python 3.9, the conversion coefficient between the photo pixel and the actual length was obtained by a scale, and then the corneal surface was fitted by arc fitting and conic curve fitting.The results of arc fitting were converted to calculate the corneal radius of curvature.The corneal eccentricity was calculated according to the general conic equation (Ax 2+ Bxy+ Cy 2+ Dx+ Ey+ F=0). The corneal asphericity was evaluated by curve fitting between the central 3-mm and the whole cornea.The use and care of the animals complied with Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.The study protocol was approved by the Ethics Committee of Shanghai Public Health Clinical Center (No.2022-A009-01). Results:The digital method of Python programming can show the corneal contour of guinea pigs completely and clearly.In the transverse plane, there was no significant difference in the corneal curvature measurements among the digital fitting in central 3-mm cornea, digital fitting in whole cornea and curvature meter ( F=1.693, P=0.190). In the sagittal plane, there was a significant difference in the corneal curvature measurements among the three methods ( F=3.500, P=0.030), and the corneal curvature measurements of the whole cornea measured by the curvature meter were significantly greater than those measured by the digital fitting ( P<0.05). There was no statistical significance in the measurements of corneal curvature radius among the three methods in the transverse plane and the sagittal plane ( F=1.817, P=0.170; F=2.050, P=0.133). The horizontal and sagittal corneal eccentricity measured by digital fitting in central 3-mm cornea were 0.55±0.15 and 0.53±0.17, which were lower than 0.66±0.10 and 0.64±0.14 measured by digital fitting in whole cornea, and the differences were statistically significant ( t=-4.860, -5.210; both at P<0.01). Conclusions:It is feasible to use Python programming digital method to measure the corneal curvature and eccentricity of guinea pigs.
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Intensive cancer treatment with drug combination is widely exploited in the clinic but suffers from inconsistent pharmacokinetics among different therapeutic agents.To overcome it,the emerging nanomedicine offers an unparalleled opportunity for encapsulating multiple drugs in a nano-carrier.Herein,a two-step super-assembled strategy was performed to unify the pharmacokinetics of a pep-tide and a small molecular compound.In this proof-of-concept study,the bioinformatics analysis firstly revealed the potential synergies towards hepatoma therapy for the associative inhibition of exportin 1(XPO1)and ataxia telangiectasia mutated-Rad3-related(ATR),and then a super-assembled nano-pill(gold nano drug carrier loaded AZD6738 and 97-110 amino acids of apoptin(AP)(AA@G))was con-structed through camouflaging AZD6738(ATR small-molecule inhibitor)-binding human serum albumin onto the AP-Au supramolecular nanoparticle.As expected,both in vitro and in vivo experiment results verified that the AA@G possessed extraordinary biocompatibility and enhanced therapeutic effect through inducing cell cycle arrest,promoting DNA damage and inhibiting DNA repair of hepatoma cell.This work not only provides a co-delivery strategy for intensive liver cancer treatment with the clinical translational potential,but develops a common approach to unify the pharmacokinetics of peptide and small-molecular compounds,thereby extending the scope of drugs for developing the advanced com-bination therapy.
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Myocardial dysfunction is the most serious complication of sepsis. Sepsis-induced myocardial dysfunction (SMD) is often associated with gastrointestinal dysfunction, but its pathophysiological significance remains unclear. The present study found that patients with SMD had higher plasma gastrin concentrations than those without SMD. In mice, knockdown of the gastrin receptor, cholecystokinin B receptor (Cckbr), aggravated lipopolysaccharide (LPS)-induced cardiac dysfunction and increased inflammation in the heart, whereas the intravenous administration of gastrin ameliorated SMD and cardiac injury. Macrophage infiltration plays a significant role in SMD because depletion of macrophages by the intravenous injection of clodronate liposomes, 48 h prior to LPS administration, alleviated LPS-induced cardiac injury in Cckbr-deficient mice. The intravenous injection of bone marrow macrophages (BMMs) overexpressing Cckbr reduced LPS-induced myocardial dysfunction. Furthermore, gastrin treatment inhibited toll-like receptor 4 (TLR4) expression through the peroxisome proliferator-activated receptor α (PPAR-α) signaling pathway in BMMs. Thus, our findings provide insights into the mechanism of the protective role of gastrin/CCKBR in SMD, which could be used to develop new treatment modalities for SMD.
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In order to explore the effect of peptidoglycan hydrolase on the viable cell counts of Bacillus amyloliquefaciens and the yield of alkaline protease, five peptidoglycan hydrolase genes (lytC, lytD, lytE, lytF and lytG) of B. amyloliquefaciens TCCC111018 were knocked out individually. The viable cell counts of the bacteria and their alkaline protease activities before and after gene deletion were determined. The viable cell counts of the knockout mutants BA ΔlytC and BA ΔlytE achieved 1.67×106 CFU/mL and 1.44×106 CFU/mL respectively after cultivation for 60 h, which were 32.5% and 14.3% higher than that of the control strain BA Δupp. Their alkaline protease activities reached 20 264 U/mL and 17 265 U/mL, respectively, which were 43.1% and 27.3% higher than that of the control strain. The results showed that deleting some of the peptidoglycan hydrolase genes effectively maintained the viable cell counts of bacteria and increased the activity of extracellular enzymes, which may provide a new idea for optimization of the microbial host for production of industrial enzymes.
Subject(s)
Bacillus amyloliquefaciens/genetics , Bacterial Proteins , Cell Count , Endopeptidases/genetics , N-Acetylmuramoyl-L-alanine Amidase/geneticsABSTRACT
Older adults have decreased control of body balance with aging and are prone to fall. As the primary point of contact between human body and ground, footwear is critical for stability of older adults. The relationship between shoe characteristics and stability of older adults was systematically reviewed to determine the effect of footwear characteristics on stability of older adults. The results show that wearing shoes with shoelaces or velcro, low heels, wide outsole and appropriate soles, or using vibrating insoles and arch support insoles can help older adults improve their stability. Excessive sole spring and low sole hardness may have adverse effects on stability of older adults. This study can provide theoretical references for older adults to choose shoes reasonably and for the manufacturers to design and make shoes to prevent falls.
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Objective:To investigate the features of median arcuate ligament syndrome (MALS) in computed tomography angiography (CTA).Methods:The retrospective and descriptive study was conducted. The clinical and imaging data of 56 patients with MALS who were admitted to Ruijin Hospital, Shanghai JiaoTong University School of Medicine from November 2019 to October 2020 were collected. There were 30 males and 26 females, aged from 19 to 78 years, with a median age of 54 years. All 56 patients underwent CTA. Observation indicators: (1) CTA examination; (2) correlation analysis; (3) surgical situations. Measurement data with normal distribution were represented as Mean± SD. Count data were described as absolute numbers or percentages. Spearman correlation analysis with test level of 0.05 was used to analyze the correlation between celiac trunk stenosis and the distance between the original location of celiac trunk and original location of superior mesenteric artery and the minimum distance of celiac trunk and superior mesenteric artery. Results:(1) CTA examination. ① Celiac trunk stenosis: results of cross sectional examina-tion of CTA showed that of 56 patients, there were 2 cases of celiac trunk occlusion, 10 cases of severe stenosis, 9 cases of moderate stenosis and 35 cases of mild stenosis. Results of sagittal examination of CTA showed that of 56 patients, there were 2 cases of celiac trunk occlusion, 21 cases of severe stenosis, 15 cases of moderate stenosis and 18 cases of mild stenosis. ② Examination of patients with different degree of vascular stenosis: according to the results of sagittal examination of CTA, the 2 cases with celiac trunk occlusion were negative for aneurysms, aortic dissection or vascular variations but positive for compensated varix of the anterior and posterior pancreatico-duodenal arches. Of the 21 cases with celiac trunk severe stenosis, 2 cases were positive for aneurysms, 1 case was positive for aortic dissection, 7 cases were positive for compensated varix of the anterior and posterior pancreaticoduodenal arches, 8 cases were positive for collateral circula-tions of anterior and posterior pancreaticoduodenal archs and 8 cases were positive for vascular variation. Of the 15 cases with celiac trunk moderate stenosis, 2 cases were positive for aneurysms, 3 cases were positive for aortic dissection, 2 cases were positive for compensated varix of the anterior and posterior pancreaticoduodenal arches, 4 cases were positive for collateral circulations of anterior and posterior pancreaticoduodenal archs and 7 cases were positive for vascular variation. Of the 18 cases with celiac trunk mild stenosis, 1 case was positive for aneurysms, 2 cases were positive for aortic dissection, 7 cases were positive for collateral circulations of the anterior and posterior pancreaticoduodenal arches, 6 cases were positive for vascular variation. All 56 patients were negative for ischemia of liver, spleen and stomach. ③ Original location of celiac trunk: of 56 patients, there were 43 cases had celiac trunk originated horizontally from the lower edge of T12 vertebral body, 2 cases had celiac trunk originated from the middle of T12 vertebral body, 1 case had celiac trunk originated from the upper part of T12 vertebral body, 7 cases had celiac trunk originated from the upper part of L1 vertebral body, 1 case had celiac trunk originated from the middle of L1 vertebral body and 2 cases had occluding celiac trunk. (2) Correlation analysis: results of sagittal observation on CTA examination showed the distance between the original location of celiac trunk and original location of superior mesenteric artery of the 56 patients was (6.0±4.0) mm. The distance between the original location of celiac trunk and original location of superior mesenteric artery of patients with celiac trunk mild, moderate or severe stenosis were (6.2±2.8)mm, (8.1±4.4)mm and (5.3±3.2)mm respectively. There were 23 cases of the 56 patients had the distance between the original location of celiac trunk and original location of superior mesenteric artery <5 mm. Results of correlation analysis showed that the degree of celiac trunk stenosis was not related to the distance between the original location of celiac trunk and original location of superior mesenteric artery ( r=?0.205, P>0.05). Results of sagittal observation on CTA examination showed the shortest distance between celiac trunk and superior mesenteric artery of the 56 patients was (3.8±2.4)mm. The shortest distance between celiac trunk and superior mesenteric artery of patients with celiac trunk mild, moderate or severe stenosis were (4.2±2.0)mm, (4.4±3.3)mm and (3.0±1.9)mm, respectively. There were 45 cases of the 56 patients had the shortest distance between celiac trunk and superior mesenteric artery <5 mm. Results of correlation analysis showed that the degree of celiac trunk stenosis was not related to the shortest distance between celiac trunk and superior mesenteric artery ( r=?0.249, P>0.05). (3) Surgical situations: of 56 patients, 2 cases were positive for clinical symptoms of abdominal pain, 54 cases were negative for clinical symptoms, 4 cases under-went surgical treatment and 52 cases not underwent surgical treatment. Of the 4 cases undergoing surgical treatment, 2 cases with abdominal pain were diagnosed as MALS by upper abdominal CTA. Celiac trunk of the 2 cases were severe stenosis and stents implantation under celiac arteriography were performed. Results of postoperative CTA showed celiac trunk was negative for obvious stenosis. The other 2 cases who were negative for clinical symptoms of MALS were planed to pancreaticoduo-denectomy for pancreatic head tumor. Results of preoperative CTA showed severe stenosis of celiac trunk and arterial bypass grafting was performed for the 2 cases during pancreaticoduodenectomy to alleviate liver ischemia caused by gastroduodenal artery ligation which would avoid the incidence of postoperative MALS associated complications. Result of postoperative CTA three-dimensional reconstruction showed the bypass vessel was unobstructed. Conclusion:Based on sagittal result of CTA examination, the stenosis of celiac trunk, the anatomical relationship between celiac trunk and arcuate ligament and the original location of celiac trunk of MALS patients can be evaluated.
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Rapid detection and response to visual threats are critical for survival in animals. The amygdala (AMY) is hypothesized to be involved in this process, but how it interacts with the visual system to do this remains unclear. By recording flash-evoked potentials simultaneously from the superior colliculus (SC), lateral posterior nucleus of the thalamus, AMY, lateral geniculate nucleus (LGN) and visual cortex, which belong to the cortical and subcortical pathways for visual fear processing, we investigated the temporal relationship between these regions in visual processing in rats. A quick flash-evoked potential (FEP) component was identified in the AMY. This emerged as early as in the LGN and was approximately 25 ms prior to the earliest component recorded in the SC, which was assumed to be an important area in visual fear. This quick P1 component in the AMY was not affected by restraint stress or corticosterone injection, but was diminished by RU38486, a glucocorticoid receptor blocker. By injecting a monosynaptic retrograde AAV tracer into the AMY, we found that it received a direct projection from the retina. These results confirm the existence of a direct connection from the retina to the AMY, that the latency in the AMY to flashes is equivalent to that in the sensory thalamus, and that the response is modulated by glucocorticoids.
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Objective:To investigate the radiosensitizing effect of Ta 4C 3-PVP nanosheets on the tumor of 4T1 murine triple-negative breast cancer cells planted in mice. Methods:4T1 tumor-bearing mice model was established by subcutaneous injection of 4T1 cells into the right flank of the female BALB/c mice. The mice were divided into four groups uniformly according to their tumor size: blank control group, Ta 4C 3-PVP group, ionizing radiation (IR) group and Ta 4C 3-PVP plus IR group. A single dose of 8 Gy X-ray local irradiation was given to xenograft tumor at 24 h after tail intravenous injection of Ta 4C 3-PVP (20 mg/kg). The xenograft tumor volume and weight, the pathological changes of tumor tissue, the expression of tumor proliferative marker Ki-67 protein, and the formation of γ-H2AX foci [a DNA double-strand breaks (DSBs) molecular marker] were detected. Tumor growth curve was established, and enhancement factor (EF) and tumor inhibition rate were calculated. Results:Compared with the blank control group, tumor growth was significantly inhibited ( t=5.41, 9.59, P < 0.05) and tumor weight was markedly decreased ( t=2.67, 4.40, P < 0.05) in both IR group and Ta 4C 3-PVP plus IR group at day 16 after IR. The EF in Ta 4C 3-PVP plus IR group was 1.57, and tumor inhibition rate in Ta 4C 3-PVP plus IR group were about 64%, which was much higher than that of IR group alone(42%). Immunohistochemistry and immunofluorescence histochemistry assays showed that the expression of Ki-67 protein was obviously decreased and the amount of γ-H2AX foci was significantly increased in both IR group and Ta 4C 3-PVP plus IR group in comparison with the blank control group ( t=5.73, 8.02, 2.97, 9.86, P < 0.05). Moreover, the inhibition of Ki-67 protein expression and the increase of γ-H2AX foci were much higher in Ta 4C 3-PVP plus IR group than that in IR group ( t=4.75, 4.42, P < 0.05). Conclusions:Ta 4C 3-PVP nanosheets could enhance radiosensitivity of xenograft tumor in 4T1 tumor-bearing mice through increasing the IR-induced DNA DSBs.
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Objective:To assess the radiosensitivity of polyvinylpyrrolidone (PVP) modified tantalum carbide (Ta 4C 3) nanosheets in human triple-negative breast cancer MDA-MB-231 cells in vitro. Methods:PVP-modified Ta 4C 3 nanosheets were synthesized and characterized. The uptake of fluoresceine isothiocyanate(FITC) labeled Ta 4C 3-PVP nanosheets in MDA-MB-231 cells was examined by fluorescent microscope. The toxicity of Ta 4C 3-PVP in the cells was measured by CCK-8 assay.These cells were divided into 4 groups: control group, Ta 4C 3-PVP group, irradiation group alone and Ta 4C 3-PVP plus irradiation group. The colony formation assay was used to evaluate the radiosensitivity. The levels of intracellular reactive oxygen species (ROS) and lipid oxidation production, malondialdehyde (MDA), were detected with an automatic microplate reader.γH2AX foci and mitotic catastrophe were observed by immunofluorescence staining. Results:Ta 4C 3-PVP nanosheets with a single-layer flaky shape were successfully synthesized. The hydrodynamic diameter and thick of Ta 4C 3-PVP nanosheets were about 143.93 nm and 1.35 nm, respectively. The FITC labeled Ta 4C 3-PVP nanosheets were uptaken by MDA-MB-231 cells, which was mainly distributed in the cytoplasm.Treatment of Ta 4C 3-PVP nanosheets at concentrations up to 400 μg/ml was noncytotoxic to MDA-MB-231 cells. The colony formation assay showed that pretreatment with Ta 4C 3-PVP at 50 and 100 μg/ml moved the survival curve of the irradiated cells downward in a concentration-dependent manner, compared to irradiation group alone. The sensitization enhancement ratio (SER D0 ) of Ta 4C 3-PVP at 50 and 100 μg/ml were 1.21 and 1.45, respectively.The intracellular ROS level in Ta 4C 3-PVP plus irradiation group was significantly higher than that in irradiation group alone at 2 and 12 h after irradiation ( q=20.01, 7.193, P<0.05). The percentage of positive cells with more than five γH2AX foci in Ta 4C 3-PVP plus irradiation group was higher than that of irradiation group alone at 1, 4 and 8 h after irradiation ( q=36.78, 14.87, 8.217, P<0.05). The content of MDA in Ta 4C 3-PVP plus irradiation group was significantly higher than that of irradiation group alone at 24 and 48 h after irradiation ( q=14.02, 7.015, P<0.05). The percentage of cells that succumbed to mitotic catastrophe in Ta 4C 3-PVP plus irradiation group was significantly higher than that in irradiation group alone at 72 h after irradiation ( q=16.33, P<0.05). Conclusions:Ta 4C 3-PVP nanosheets could enhance the radiosensitivity of human triple-negative breast cancer MDA-MB-231 cells through an increase of the intracellular ROS induced by irradiation.
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1,3-1,4-β-glucanase (E.C.3.2.1.73) is an important industrial enzyme which cleave β-glucans into oligosaccharides through strictly cutting the β-1,4 glycosidic bonds in 3-O-substituted glucopyranose units. Microbial 1,3-1,4-β-glucanase belongs to retaining glycosyl hydrolases of family 16 with a jellyroll β-sandwich fold structure. The present paper reviews the industrial application and protein engineering of microbial β-glucanases in the last decades and forecasts the research prospects of microbial β-glucanases.