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BACKGROUND:A large number of studies have confirmed that tissue engineering scaffolds can almost completely repair osteochondral defects.However,when osteochondral defects are complicated with infection,even after thorough debridement in the early stage,the repair effect of simple osteochondral tissue engineering scaffolds is often unsatisfactory. OBJECTIVE:To prepare fibroin/chitosan/nano-hydroxyapatite scaffold loaded with vancomycin hydrochloride sustained release microspheres,and to investigate the repair effect on infected osteochondral defect in distal femur of rabbit. METHODS:(1)Vancomycin hydrochloride sustained release microspheres were prepared by emulsified solvent evaporation method.The sustained-release microspheres of different weights(7.5,10,and 12.5 mg)were mixed with fibroin protein-chitosan nanohydroxyapatite solution,and the scaffolds of fibroin protein/chitosan/nano-hydroxyapatite were prepared by chemical crosslinking method.The porosity,water absorption and expansion rate,hot water loss rate of the scaffolds,and drug sustained-release in vitro were characterized.(2)Forty-five New Zealand white rabbits were randomly divided into blank group,control group,and experimental group,with 15 rabbits in each group.The osteochondral defect and infection model of the distal femur of the right hind limb was established in both groups.The blank group was not treated,and the control group was implanted with fibroin protein-chitosan-nano-hydroxyapatite scaffold.Vancomycin hydrochloride sustained-release microspheres(10 mg)of fibroin/chitosan/nano-hydroxyapatite scaffold were implanted in the defect of the experimental group.The levels of C-reactive protein and leukocytes in blood samples were detected 1 week after operation.At 4,8 and 12 weeks after operation,the tissue of the operative area was taken for gross observation and pathological observation. RESULTS AND CONCLUSION:(1)With the increase of sustained-release microspheres content,the porosity of scaffolds decreased,and there was significant difference among groups(P<0.05).There were no significant differences in the pore size,water absorption expansion rate and hot water loss rate among the three groups(P>0.05).Vancomycin hydrochloride was released sustainably in vitro for more than 30 days in all three groups of scaffolds.(2)The levels of C-reactive protein and leukocytes in blood samples of the experimental group were lower than those of the blank group and control group(P<0.05).The repair of gross cartilage in the experimental group was significantly better than that in the blank group and the control group.Hematoxylin-eosin,Masson,Alcian blue and type Ⅱ collagen immunohistochemical stainings showed that the osteochondral repair effect of the experimental group was significantly better than that of the blank group and the control group at each time point.(3)The results showed that fibroin/chitosan/nano-hydroxyapatite scaffolds loaded with vancomycin hydrochloride sustained-release microspheres could effectively promote the repair of open osteochondral defects.
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The parabrachial nucleus (PBN) integrates interoceptive and exteroceptive information to control various behavioral and physiological processes including breathing, emotion, and sleep/wake regulation through the neural circuits that connect to the forebrain and the brainstem. However, the precise identity and function of distinct PBN subpopulations are still largely unknown. Here, we leveraged molecular characterization, retrograde tracing, optogenetics, chemogenetics, and electrocortical recording approaches to identify a small subpopulation of neurotensin-expressing neurons in the PBN that largely project to the emotional control regions in the forebrain, rather than the medulla. Their activation induces freezing and anxiety-like behaviors, which in turn result in tachypnea. In addition, optogenetic and chemogenetic manipulations of these neurons revealed their function in promoting wakefulness and maintaining sleep architecture. We propose that these neurons comprise a PBN subpopulation with specific gene expression, connectivity, and function, which play essential roles in behavioral and physiological regulation.
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Núcleos Parabraquiales/fisiología , Vigilia/fisiología , Neuronas/fisiología , Emociones , SueñoRESUMEN
OBJECTIVE@#To retrospectively analyze the results of chromosomal microarray analysis (CMA) and parental origins of unbalanced translocations among 17 patients, so as to provide reference for their genetic counseling.@*METHODS@#The results of CMA for 7 001 samples tested in Chengdu Women and Children's Central Hospital from January 2019 to January 2022 were retrospectively reviewed. Unbalanced reciprocal translocation was defined as two non-homologous chromosomes with lost and gained segments respectively or both with gained segments, and their parental origins were identified by parental chromosomal karyotyping and/or fluorescence in situ hybridization (FISH).@*RESULTS@#In total 17 unbalanced translocations were identified. In three cases, two non-homologous chromosomes both had gained segments, which constituted a derivative chromosome, with the total number of chromosomes being 47. In the remaining 14 cases, there was a terminal deletion on one chromosome and a terminal duplication on the other, 10 of which were confirmed by karyotyping, with the total number of chromosomes being 46. In the derivative chromosome, the lost segment was replaced by a gained segment from another chromosome. Among 15 cases undergoing parental origin analysis, 12 had paternal or maternal chromosomal abnormalities, including 11 balanced translocations and 1 unbalanced translocation. The unbalanced gametes therefore may form through meiosis. In 3 cases, the parental chromosomes were normal, indicating a de novo origin.@*CONCLUSION@#Discovery of terminal duplication and deletion or gained segments on two non-homologous chromosomes by CMA is suggestive of parental balanced translocation, which can facilitate genetic counseling and assessment the recurrence risk for subsequent pregnancies.
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Niño , Embarazo , Humanos , Femenino , Hibridación Fluorescente in Situ , Estudios Retrospectivos , Translocación Genética , Análisis por Micromatrices , CromosomasRESUMEN
Laparoscopic hepatectomy has been routinely used in all tumor types of the liver, but for tumors in the Ⅵ, Ⅶ and Ⅷ segments of the liver (referred to as tumors in a special position of the liver in this paper), laparoscopic hepatectomy has some difficulties in aspect of vision and operation due to the deep anatomical position. In the process of continuous experience accumulation, hepatobiliary surgeons have explored different surgical positions, surgical methods and surgical paths to solve the encountered difficulties. For tumors located in a special position of the liver, conventional laparoscopy, change of surgical posture, thoracoscopic or combined laparoscopy and retroperitoneal laparoscopy are effective and feasible. All of them have their own advantages.
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In order to explore a suitable pathway for the scientific and technological achievement transformation at prefecture-level public hospitals, in October 2020, a prefecture-level tertiary hospital carried out the " 3+ 3+ 3" mode for scientific and technological innovation and achievement transformation. A three-level management structure was established, consisting of the ministry of science and education, the working group on the transformation of scientific and technological achievements, and the leading group. Three management systems were improved and formulated, including the " measures for reimbursement and rewards of scientific and technological achievements ", the " measures for intellectual property management", and the " implementation plan for promoting the transfer and transformation of scientific and technological achievements". The management measures for the three stages of intellectual property protection, incubation of scientific and technological achievements, and transfer and transformation of achievements were improved. These measures provided organizational support, institutional support, and feasible paths for this practice. After nearly three years of practice, hospitals had reduced the number of low-quality patent applications, and the number and amount of scientific and technological achievements transformed increased from 1 achievement and 10 000 yuan in 2020 to 9 achievements and 320 000 yuan in 2022. This exploration could provide a reference for the transfer and transformation of professional scientific and technological achievements in prefecture-level public hospitals in China.
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The triple-negative breast cancer has a relatively poor prognosis with limited therapeutic options. This subtype is highly immunogenetic and exhibits rich tumor infiltrated lymphocytes in the tumor microenvironment. However, immunotherapy alone is less effective as compared with the doublet of chemotherapy and immunotherapy in TNBC. The efficacy in early recurrence settings of mTNBC exceeds that in heavily treated subgroups. Meanwhile, other combinations including anti-angiogenesis, immune modulators, and PARPi elicit a promising effect. Herein, this paper reviews the progress of efficacy, safety, and the outlook in the immunotherapy of TNBC disease.
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Purpose@#RIOK1 has been proved to play an important role in cancer cell proliferation and migration in various types of cancers—such as colorectal and gastric cancers. However, the expression of RIOK1 in breast cancer (BC) and the relationship between RIOK1 expression and the development of BC are not well characterized. In this study, we assessed the expression of RIOK1 in BC and evaluated the mechanisms underlying its biological function in this disease context. @*Materials and Methods@#We used immunohistochemistry, western blot and quantitative real-time polymerase chain reaction to evaluate the expression of RIOK1 in BC patients. Then, knockdown or overexpression of RIOK1 were used to evaluate the effect on BC cells in vitro and in vivo. Finally, we predicted miR-204-5p could be a potential regulator of RIOK1. @*Results@#We found that the expression levels of RIOK1 were significantly higher in hormone receptor (HR)–negative BC patients and was associated with tumor grades (p=0.010) and p53 expression (p=0.008) and survival duration (p=0.011). Kaplan-Meier analysis suggested a tendency for the poor prognosis. In vitro, knockdown of RIOK1 could inhibit proliferation, invasion, and induced apoptosis in HR-negative BC cells and inhibited tumorigenesis in vivo, while overexpression of RIOK1 promoted HR-positive tumor progression. MiR-204-5p could regulate RIOK1 expression and be involved in BC progression. @*Conclusion@#These findings indicate that RIOK1 expression could be a biomarker of HR-negative BC, and it may serve as an effective prognostic indicator and promote BC progression.
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Bovine ephemeral fever virus (BEFV) causes bovine ephemeral fever, which can produce considerable economic damage to the cattle industry. However, there is limited experimental evidence regarding the underlying mechanisms of BEFV. Annexin A2 (AnxA2) is a calcium and lipid-conjugated protein that binds phospholipids and the cytoskeleton in a Ca 2+ -dependent manner, and it participates in various cellular functions, including vesicular trafficking, organization of membrane domains, and virus proliferation. The role of the AnxA2 gene during virus infection has not yet been reported. In this study, we observed that AnxA2 gene expression was up-regulated in BHK-21 cells infected with the virus.Additionally, overexpression of the AnxA2 gene promoted the release of mature virus particles, whereas BEFV replication was remarkably inhibited after reducing AnxA2 gene expression by using the small interfering RNA (siRNA). For viral proteins, overexpression of the Matrix (M) gene promotes the release of mature virus particles. Moreover, the AnxA2 protein interaction with the M protein of BEFV was confirmed by GST pull-down and coimmunoprecipitation assays. Experimental results indicate that the C-terminal domain (268– 334 aa) of AxnA2 contributes to this interaction. An additional mechanistic study showed that AnxA2 protein interacts with M protein and mediates the localization of the M protein at the plasma membrane. Furthermore, the absence of the AnxA2-V domain could attenuate the effect of AnxA2 on BEFV replication. These findings can contribute to elucidating the regulation of BEFV replication and may have implications for antiviral strategy development.
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Objective To evaluate the effects of preoperative prophylactic infusion of cephalosporins antibiotics on electrocardiogram(ECG)during anesthesia induction with propofol. Methods Fifty female patients,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 20-50 yr,weighing 43-75kg,scheduled for elective surgery,were divided into cefuroxime sodium group(group CEFU)and cefathiamidine group(group CEFA),with 25 patients in each group. After cefuroxime sodium was infused in group CEFU or cefathiamidine was infused in group CEFA,propofol was given by target-controlled infusion(plasma concentration 4 μg/ml).Before infusion of cefuroxime sodium or cefathiamidine,after completion of infusion and at 5min after propofol reached the target concentration,12-lead ECG was recorded,and mean arterial pressure and heart rate were simultaneously recorded. Tp-e interval and Tp-e/QT ratio were calculated.Results Compared with the baseline before infusion of cefuroxime sodium or cefathiamidine,Tp-e interval was significantly prolonged after completion of infusion and at 5min after propofol reached the target concentration in group CEFU,and Tp-e interval was significantly prolonged,and Tp-e/QT ratio was enlarged in group CEFA(P0.05).Conclusion Preoperative prophylactic infusion of cefuroxime sodium produces less ECG interference than preoperative prophylactic infusion of cefathiamidine during anesthesia induction with propofol.
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Objective To discover more novel bat viruses and molecularly characterize bat-borne bocavirus diversity in Yunnan.Methods Twenty-six Aselliscus stoliczkanus were sampled in Jinghong , Yunnan, and subjected to viral metagenomic analysis.Specific PCR was used to detect any bocavirus in these samples based on the metagenomic result , while full genome was amplified and compared with other bocaviruses .Results and Conclusion Totally, 3 of the 26 (11.5%) bats were positive for bocavirus, the full genome of which contained 5203 nucleotides and could encode NS1, NP and VP1/VP2 proteins.Phylogenetic analysis showed that this virus shared up to 58.7% and 53.3% amino acid identities with canine bocavirus 1 and canine minute virus .According to ICTV criteria (85%amino acid sequence identity ) on a new species of bocavirus , this virus could be a novel species within genus Bocaparvovirus .This study provides important data to better understand viral diversity in bats and to uncover the relationship between bocavirus and its hosts .
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Objective To investigate the influence of low tidal volume (VT ) combined with low level positive end-expiratory pressure(PEEP) on the respiratory function of elderly patients in general anesthesia .Methods 46 elderly patients scheduled for up-per abdominal surgery in our hospital were randomly divided into two groups ,23 cases in each group .The control group received the conventional tidal volume (VT ) ,while the observation group was given low VT combined with 3cm H2 O of PEEP ,both groups were given intermittent positive pressure ventilation (IPPV) .The arterial blood was collected for conducting the blood gas analysis before operation(T0 ) ,at 30 min after anesthetic intubation(T1 ) and 15 min after extubation(T2 ) .The changes of the indexes of PaO2 ,PaCO2 ,Ppeek and A-aDO2 at various time points were compared between the two groups .Results PaO2 at T2 in the observa-tion group was(83 .58 ± 2 .43) mmHg ,which was significantly higher than that in the control group ,while A-aDO2 in the observa-tion group was(21 .50 ± 2 .12) ,which was significantly lower than that in the control group(P<0 .05);Ppeek at T1 in the observa-tion group was(11 .27 ± 1 .22) cm H2O ,which was significantly lower than that in the control group(P<0 .05) .Conclusion The application of low VT combined with low PEEP in general anesthesia can effectively improve postoperative hypoxemia in elderly pa-tients and is conducive to the respiratory function recovery ,which has better clinical application value .
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Objective To explore the feasibility and clinical value of the trans-umbilical single-port laparoscopic operation with routine apparatus in treatment of hepatobiliary diseases. Methods From Mar 2010 to Sep 2012, 415 patients with hepatobiliary disease, splenic disease, abdominal cavity disease or combine hysteromyoma were performed by trans-umbilical single-port laparoscopic operation with routine apparatus. The clinical data of 415 cases were analyzed respectively.Result All patients except one patients who was added one port for hysterectomy, were performed successfully trans-umbilical single-port laparoscopic operation with routine apparatus without conversion to laparotomy or conventional laparoscopic surgery. Conclusions The trans-umbilical single-port laparoscopic operation with routine apparatus is safe and effective in treatment of hepatobiliary diseases. Limited by the anatomy position, the trans-umbilical single-port laparoscopic operation with routine apparatus can cure most hepatobiliary diseases. So the operator must have the experience both in single-port laparoscopic operation and traditional laparoscopic operation. With the improvement of rotatable equipment, the practical procedure is expected to be generalized in the future.
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Objective To explore a new technique to treat the benign stricture at hilar bile duct of plastic anastomosis, so as to obtain the theoretical basis and the feasibilities of practical application in treating the benign stricture at hilar bile duct with plastic anastomosis through the animal experiments. Methods 30 miniature pigs were randomly divided into three groups. Group A (control group):2 centimeters of the bile duct above the duodenum was isolated;Group B:The same separation of bile duct with group A, then making physical injury on it with the clamps and electric heat;Group C:Making bile duct injury model in group C as group B,and then monitoring of the diet, feces, urine, ALT, AST and bilirubin etc. When the bile duct stricture was formed, taking the plastic anastomosis operation in this group. After all the operations, we observed the diet, mental state and the color of the urine of animals in all the three groups, and tested ALT,AST,T-BIL and D-BIL levels on the pre- and post-day and every 7 days after surgery respectively. After three months of the surgery,we executed all the pigs,picked up part of the liver tissue,then preserved them by liquid nitrogen for pathological examination. Results From the day before operative-day to the 30 days after operation, there was no significant change in ALT, AST, T-BIL and D-BIL in group A,while the relative indicators of group B and C had obvious changes ( <0.05) . The indicators of group B were significantly increased 21 days after surgery, compared with pre-operation and 14 days after operation respectively ( <0.05) . In group C, the indicators were significantly declined 14 to 21 days after the plastic anastomosis compared with pre-operation (<0.05) . Conclusions Treating benign stricture at hilar bile duct of miniature pigs by plastic anastomosis is feasible and practicable. This study provides an experimental basis for clinical application of plastic anastomosis in treatment of benign stricture at hilar bile duct.
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ObjectiveTo study the best extractive process for Vaccinium Fragile Franch. Methods Single factor and orthogonal experiments were designed to determine the extractive rate as the index of evaluation and to optimize the extraction technology. ResultsThe experiments showed that adding 9 times of 70% methanol, refluxing for 2 times, each times 2 hours at the 65℃C was the best extractive process.ConclusionThe process was stable and reasonable, which formed the scientific and reasonable basis for developing the active parts from Vaccinium Fragile Franch.
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Objective To explore the optimal conditions for extraction of Polygala fallax Hemsl. Methods Various trials were conducted to extract P. Fallax Hemsl. With L_(16) (4~5) orthogonal experiments. Results The optimal experimental parameters for extraction were 95% ethenol, dosage-liquor ration at 1:5, three-times extraction at 75℃, 1.5 hours for each extraction, and extraction rate at 11.89%. Conclusion Orthogonal test is convenient and accurate in extracting P. Fallax Hemsl.
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This paper summarized the recent researches of chemistry compounds of Luoxiao and pointed out that Oxyria digyna (L.) Hill and Oxyria sinensis Hemsl.Flavonoid were the main components of it, with Flavonoid compounds had extensive pharmacological activity.
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<p><b>OBJECTIVE</b>To study the chemical constituents of the roots of Osbeckia opipara.</p><p><b>METHOD</b>Repeated column chromatography over silica gel, RP-18 and Sephadex LH-20, and preparative thin layer chromatography(PTLC) were used to isolate the compounds, whose structures were determined by spectroscopic methods by direct comparing spectral data with those reported references.</p><p><b>RESULT</b>From the MeOH extract of the roots O. opipara, twelve compounds were isolated and identified as follows: lasiodiplodin (1) , de-O-methyllasiodiplodin (2), 2, 3- dihydro-2-hydroxy-2, 4-dimethyl-5-trans-propenylfuran-3-one (3), integracin (4), 5alpha, 8alpha-epidioxy-(22E, 24R)-ergosta-6, 22-dien-3beta-ol (5), 3, 3', 4'-tri-O-methylellagic acid (6), 5-hydroxymethyl furaldehyde (7), vomifolio (8) , betulintic acid (9), 2alpha-hydroxyursolic acid (10), (24R)-stigmast-4-ene-3-one (11), and eugenitin (12).</p><p><b>CONCLUSION</b>Compounds 1-12 were isolated from O. opipara for the first time.</p>
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Colestenonas , Química , Fermentación , Cuerpos Fructíferos de los Hongos , Química , Melastomataceae , Química , Estructura Molecular , Extractos Vegetales , Química , Raíces de Plantas , Química , Análisis Espectral , Triterpenos , QuímicaRESUMEN
A xylosidase gene, labeled as BH1068 in genome of Bacillus halodurans C-125, was successfully cloned and overexpressed in Escherichia coli JM109. The purified enzyme was thoroughly characterized and its xylosidase function was unambiguously confirmed. It has maximum activities in neutral condition and is stable over a wide range of pH (4.5-9.0). The enzyme has a broad temperature optimal (35 degrees C-45 degrees C) and is quite stable at temperature up to 45 degrees C. The unique pH and temperature profiles of the enzyme should allow a wide range of xylanolytic operational conditions. With high specific activity of 174 mU/mg protein for its artificial substrate (p-nitrophenyl-beta-xylose) and low xylose inhibition (inhibitor constant Ki = 300 mmol/L), this enzyme is among the most active and high tolerant bacterial xylosidase to xylose inhibition. Its high synergy with commercial xylanase has been demonstrated with beechwood xylan hydrolysis, achieving a hydrolysis yield of 40%. Its neutral pH optimal and high tolerance to product inhibition complements well with its fungal counterparts that are only optimal at acidic pH and susceptible to xylose inhibition. In conclusion, this enzyme has high potential in the saccharification of xylan and xylan-containing polysaccharides.
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Secuencia de Aminoácidos , Bacillus , Clasificación , Genética , Clonación Molecular , Escherichia coli , Genética , Metabolismo , Hidrólisis , Datos de Secuencia Molecular , Proteínas Recombinantes , Genética , Metabolismo , Especificidad por Sustrato , Xilosa , Metabolismo , Xilosidasas , Genética , MetabolismoRESUMEN
Polygonum bistorta L.in China was used as traditional Chinese medicine.The chemical constituents and biological activity studies on this species were summarized and reviewed.
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Objective To express and purify pET-p53 fusion protein and investigate the effects of the protein on the proliferation of human leukemia cell line K562. Methods The fragment of human wild type p53 cDNA was amplified by PCR and the expression plasmid of pET-p53 was constructed. Recombinant plasmids were transformed into E.coli BL21,then induced by IPTG at 1mmol/L. The protein was purified by the column of Ni-NAT and analyzed by SDS-PAGE. After treated with purified P53 protein with different concentrations,the proliferation of K562 was tested by MTT assay, clone formation and FCM. Results Prokaryotic expression vectors of pET-p53 were constructed correctly. pET-p53 fusion protein was successfully expressed and purified. With its increasing concentrations, P53 protein reversed its effect on K562 significantly. The ratio of inhibition had linear relation to the concentration of P53 protein when the concentration of the protein was between 0.1?g/ml and 100?g/ml. And its IC50 was 6.74?g/ml. Conclusion The obtained pET-p53 fusion protein might induce the leukemia cell line K562 apoptosis.