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Objective To screen out specific aldosterone(ALD)antibodies using phage display technology and recom-binant antibody technology,providing raw materials for the research and development of ALD diagnostic kits.Methods Five healthy and clean New Zealand white rabbits were selected and immunized for the first time against the diluted ALD-keyhole limpet hemocyanin antigen(2 mg·L-1)using a multi-point injection method on the back,with a dose of 1 mg per rabbit.Immunization was administered again every 2 weeks,with a 50%reduction in dose.Starting from the third immunization,the ear vein blood of the rabbits was collected one week after each immunization.A chemiluminescent plate coated with 0.25 mg·L-1 ALD-bovine serum albumin antigen was used to measure serum titers via indirect and competitive methods.After the 5th immunization,the rabbit with high serum titers and good specificity was selected,and its spleen and bone marrow were removed.The spleen tissue was grinded,and RNA was extracted using TRIzol reagent in one step to obtain gene sequences in the variable region of light chain(VL)and the variable region of heavy chain(VH).The single-chain variable fragment(ScFv)was connected through the linker and constructed into the bacteriophage vector Pcomb3xss;then,it was carried to Escherichia coli TG1 through electrotransformation,and the ALD ScFv phage display library was constructed accordingly.Three to five rounds of enrichment screening were performed against the library.Monoclonal clones,identified by enzyme-linked immunosorbent assay(ELISA)competitive method,were selected for phage supernatant preparation,and a highly competitive clone sequence was obtained.The screened clone sequence was inserted into the pCMV3 expression vector,and the HEK293 cell was transfected using the transient transfection method after the plasmid was extracted.One week later,the supernatant was collected,and its purity and expression were identified by affinity chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Results After the 5th immunization,the serum titers of 5 rabbits were indirectly tested,and the results showed that the serum titers of 4# and 5# white rabbits were still greater than 10,000 after being diluted by 32,000 times.The test results based on the competitive method showed that the ratio of low to high values in the plasma sample of 5#white rabbit was 2∶1,superior to that of other white rabbits.The 5# white rabbit was selected for phage library construction.The VL and VH gene fragments were amplified by conventional polymerase chain reaction,and then bridged into ScFv(VL+VH).The agar gel electrophoresis analysis showed that the size of the band was about 750 bp,which was consistent with the size of the originally designed fragment.ScFv was cleaved and electroporated into Escherichia coli TG1 to construct a phage library with a storage capacity of 4.73 × 108 cfu·mL-1.After 3 rounds of washing,300 monoclonal clones were selected from the outbound petri dishes to prepare monoclonal bacteriophages.The ELISA results showed a positive rate of 100%among the 300 clones,and 42 clones were tested positive for calibration competition,with a screening rate of 14%.The 42 positive clones were further subjected to clinical sample competition testing,and 16 monoclonal strains that met the requirements were screened.The 16 strains were retested,and the results of the two tests were consistent.After sequencing,6 antibody sequences were selected for construction and expression.After purification,SDS-PAGE reduced gel electrophoresis results showed that there were bands at positions 50,000 on the heavy chain and 25,000 on the light chain.Six highly affinitive and competitive rabbit ALD monoclonal antibodies were obtained.Conclusion Six highly affinitive and competitive rabbit ALD antibodies are successfully screened using phage display technology,which provides a reference method for the discovery of small molecule antibodies.The screened AD1 85 and AD277 antibodies show a competitive advantage twice that of the positive control in the competition of calibration and clinical samples,providing a possibility for the development of raw materials for ALD detection kits.
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Objective To study the effects of different pellet feed hardness on the growth and reproduction, feed utilization rate, and environmental dust in laboratory mice.Methods One hundred of fifty 50 3-week-old SPF-grade C57BL/6JGpt and 150 ICR laboratory mice were randomly divided into three groups, with an equal number of males and females. They were fed diets with different hardness of 18.62 kg, 23.15 kg, and 27.89 kg. Body weight, feed utilization rate, and dust levels in cages were recorded and calculated for mice aged 3-10 weeks. Forty-five 6-week-old male mice and ninety 4-week-old female mice from each strain were randomly divided into three groups and fed pellet feeds with three different hardness levels. After 2 weeks of adaptation to the same hardness feed, the mice were paired at a 1∶2 male-to-female ratio and monitored for reproductive data for 3 months.Results At the age of 4 weeks, the body weight of male C57BL/6JGpt mice in 23.15 kg group was significantly higher than that in the 18.62 kg and 27.89 kg groups (P<0.01), and the body weight of females in the 18.62 kg group was significantly higher than that in the 27.89 kg group (P<0.05). There was no significant difference in body weight among ICR mice aged 3-10 weeks across different feed hardness groups (P>0.05). For both strains, feed utilization rate for males was higher than that for females across different feed hardness groups at all weeks of age (P<0.01). Compared to the 27.89 kg group, both the 18.62 kg and 23.15 kg groups showed a significant increase in the 50-mesh dust levels in cages for both strains aged 4-8 weeks (except for 7-week-old C57BL/6JGpt mice) (P<0.05). For both C57BL/6JGpt and ICR mice, there was no significant difference in basic reproductive performance such as interval between the first litter and the monthly production index among the three feed hardness groups during the experimental period (P>0.05). However, the monthly production index of C57BL/6JGpt mice first increased and then decreased with the increase of feed hardness, while that of ICR mice increased with increasing feed hardness, though these differences were not statistically significant (P>0.05).Conclusion Different strains and genders had different tolerance to feed hardness. C57BL/6JGpt mice are more adapted to lower hardness feeds, while ICR mice are better suited to slightly higher hardness feeds.
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Rehabilitation and multidisciplinary team management are very important to improve the prognosis of children with prolonged mechanical ventilation(PMV). The current status of rehabilitation intervention including physical therapy, neuromuscular electrical stimulation and inspiratory muscle training in adult patients with PMV were discussed, aiming to provide some evidence for the implementation of rehabilitation in children with PMV.
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Objective:To explore the effects of external diaphragm electrical stimulation on the diaphragm thickness and function in mechanically ventilated children.Methods:A randomized controlled trial was conducted in children who were admitted to PICU at Children′s Hospital of Fudan University and received mechanical ventilation between June 2021 and April 2022.The control group was given the routine treatment of mechanical ventilation, and the intervention group was given external diaphragm electrical stimulation in the early stage of mechanical ventilation in addition to routine treatment.Diaphragm thickness was continuously measured by bedside ultrasound every day for one week after mechanical ventilation, and the changing trend of diaphragm thickness was observed, and the diaphragmatic thickening fraction (DTf) and the incidence of ventilator-induced diaphragmtic dysfunction(VIDD) were calculated at the same time.Results:A total of 32 valid samples were included, including 15 cases in intervention group (10 males) and 17 cases in control group (11 males). The median age of the patients was 33 (10, 77) months, and the median duration of mechanical ventilation was 12 (8, 21) days.The reasons for mechanical ventilation in children included respiratory insufficiency in ten cases, brain dysfunction in ten cases, heart failure in eight cases, and postoperative surgery in four cases.The diaphragm end-expiratory thickness (DTe) in intervention group and the control group showed a gradually decreasing trend from the 1st day to the 7th day.The left thickness was reduced by 11% on the 7th day compared to 1st day in intervention group, which was reduced by 18% in control group; the average daily DTe was reduced by 2% per day in intervention group and by 3% per day in control group.The trends on the right and left were similar.The DTe thickness in the intervention group was greater than that in control group, among which, the mean DTe thickness in the left side of the intervention group on the 7th day was (0.110 7±0.023 7)cm, which was greater than that in control group (0.093 5±0.016 9)cm, and the difference was statistically significant ( t=-2.372, P<0.05); On the second day, the mean DTe thickness on the right side in the intervention group was (0.1267±0.0277) cm, which was greater than that in control group (0.104 7±0.018 1)cm, and the difference was statistically significant ( t=-2.688, P<0.05). DTf in the intervention group was lower than that in control group at 7th day, but the difference was not statistically significant(left DTf: adjusted mean difference was -0.117, P=0.088; right DTf: adjusted mean difference was -0.065, P=0.277). The incidence of VIDD in the intervention group was lower than that in control group(33.3% vs.41.2%), but the difference was not statistically significant ( χ2=0.005, P=0.946). Conclusion:External diaphragmatic electrical stimulation may be helpful for alleviating diaphragmatic atrophy in mechanically ventilated children.However, whether the improvement of diaphragm atrophy is beneficial to clinical outcome still needs further study.
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Objective:To study and analyze the application effect of intervention plan based on unplanned readmission risk model (LACE) in the rehabilitation of patients with acute myocardial infarction (AMI) complicated with cardiac shock (CS) after percutaneous coronary intervention (PCI).Methods:Ninety-three patients with AMI complicated with CS who received PCI in Tianjin Union Medical Center from January 2019 to December 2020 were enrolled. The patients were divided into LACE intervention group ( n = 46) and routine intervention group ( n = 47) according to the different nursing intervention methods. The patients in the routine intervention group received routine interventions, including drug care, diet care, psychological care, health education and telephone follow-up, while those in the LACE intervention group were assessed for the risk of LACE, and then intervention measures were formulated according to the score of LACE index, including strengthening risk awareness, life behavior, daily life ability, self-care ability, health recovery and health needs. The follow-up period in both groups was 3 months. The changes of cardiac function, incidence of adverse cardiac events, readmission rate, self-management ability after intervention and quality of life level before and after intervention were compared between the two groups. Results:There was no significant difference in cardiac function or quality of life before intervention between the two groups. After intervention for 3 months, the cardiac function and quality of life in the two groups were improved as compared with those before intervention. The left ventricular ejection fraction (LVEF) in the LACE intervention group was significantly higher than that in the routine intervention group (0.533±0.076 vs. 0.492±0.072, P < 0.05), the left ventricular end diastolic diameter (LVEDD) was significantly lower than that in the routine intervention group (mm: 47.09±7.01 vs. 53.23±7.15, P < 0.01), and the World Health Organization Quality of Life-brief (WHOQOL-BREF) score was also significantly higher than that in the routine intervention group (66.32±6.19 vs. 55.79±7.26, P < 0.01). The scores of self-management ability in the coronary heart disease self-management scale (CSMS) after intervention in the LACE intervention group were significantly higher than those in the routine intervention group (adverse hobbies score: 17.37±3.24 vs. 14.21±2.73, symptoms score: 14.82±3.11 vs. 10.56±2.65, emotional cognition score: 16.17±2.83 vs. 12.95±2.41, first aid score: 11.85±1.94 vs. 10.62±1.56, disease knowledge score: 15.58±2.73 vs. 12.68±2.61, daily life score: 17.80±2.61 vs. 14.33±2.36, treatment compliance score: 11.47±1.84 vs. 8.56±1.23, all P < 0.01). The incidence of adverse cardiac events and readmission rate in the LACE intervention group were significantly lower than those in the routine intervention group (10.87% vs. 29.79%, 4.35% vs. 17.02%, both P < 0.05). Conclusion:The intervention plan based on LACE risk model can effectively promote postoperative rehabilitation of patients with AMI complicated with CS after PCI, and also help to improve patients' self-management ability and quality of life, which is worthy of clinical promotion and application.
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Objective:To analyze the dimensional structure and the item characteristics of the Chinese version of the cerebral palsy quality of life questionnaire for children (CPQOL-child).Methods:Multi-dimensional and single-dimensional item response models were used. Ninety children aged 4 to 12 with cerebral palsy were evaluated using the CPQOL-child. The internal consistency was determined by analyzing the correlations among the different dimensions using a multi-dimensional response model. Dimensions were merged or split based on a one-dimensional Rasch analysis of the primary and secondary dimensions to revise the main and additional questionnaires.Results:The multi-dimensional item response model analysis found greater than 0.90 correlation among the four dimensions of social well-being and tolerance, function, participation ability and physical health, and emotional health and self-esteem, showing that they have good internal consistency and could be analyzed as the primary dimension. The correlation coefficients relating the other three dimensions were relatively low and could be analyzed as secondary dimensions. A single-dimension Rasch analysis was used to form a revised CPQOL main questionnaire consisting of 40 items, with three additional questionnaires for pain and disability impact consisting of 8 items, for access to services consisting of 6 items, and for family health consisting of 3 items. Compared with the original 65 items, the 8 items deleted were mainly about relationships between children and with siblings, providing respite services, as well as community and school services.Conclusions:Applying the Chinese version of the CPQOL-Child in Shanghai requires appropriate adjustments based on the children′s cultural background.
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Objective:To explore the reproducibility of ultrasound measurements of children′s anterior, middle and posterior diaphragm motions.Methods:Thirty children admitted to a pediatric intensive care unit were positioned supine and a 5MHz ultrasound probe was placed over the intersection of their right midclavicular line with the costal margin. M-mode ultrasound was used to record the excursion and contraction velocity of the anterior, middle and posterior diaphragm during respiration. The observations were duplicated so the repeatability of the measurements could be evaluated using intra-group correlation coefficients calculated for the diaphragm excursions and the contraction velocities. Analysis of variance was used to explore the differences in excursion and contraction velocity among different parts of the diaphragm.Results:The intra-group correlation coefficients calculated for the anterior, middle and posterior diaphragm were 0.89, 0.95 and 0.90 respectively. The corresponding values for the contraction velocities were 0.90, 0.94 and 0.95 respectively. Both variables measured by ultrasound showed high repeatability. The average anterior, middle and posterior diaphragm excursion values (in mm) were 8.1±3.1, 7.4±3.0 and 5.5±2.3, and the corresponding average contraction velocities (in mm/s) were 12.5±4.8, 11.5±6.3 and 8.9±4.0.Conclusions:Measurements of children′s diaphragm motions using ultrasound show high repeatability. The excursions and contraction velocities of the anterior, middle and posterior diaphragm differ in children. The motion of one part of the diaphragm cannot represent the functioning of the entire diaphragm.
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Objective@#To develop the early respiratory rehabilitation techniques in pediatric intensive care unit(PICU) for children with mechanical ventilation.@*Methods@#A respiratory rehabilitation team was established, and early respiratory rehabilitation intervention methods for critically ill children were also developed.Early bedside rehabilitation were implemented after the vital signs of mechanical ventilation children were stable for 24 hours, and the rehabilitation effect and adverse events were observed.@*Results@#According to the patient′s condition, 23 patients in PICU received early respiratory rehabilitation after incubations, including 10 males and 13 females, aged 57.0(17.5, 91.5) months.There was no adverse event in 729 rehabilitation practices.The average consulting time was 8.0(4.0, 18.0) days after incubations.The average incubation time was 693.5(345.2, 1 027.1)hours of all the patients.As to 16 patients who were weaned before charged out, the average incubation time was 566.2(243.4, 928.7)hours, and the average incubation time from respiratory rehabilitation performance to weaning was 168.0(6.0, 564.0) hours.In 19 patients who only received limbs massage and passive movements, the average incubation time was 381.7(309.6, 541.4) hours and the average incubation time from respiratory rehabilitation performance to weaning was 187.2(81.5, 382.9) hours.However the difference between these who received respiratory rehabilitation and only received limbs massage and passive movements was not statistically significant.@*Conclusion@#It′s safe and feasible to perform the early respiratory rehabilitation in PICU for critically ill children with mechanical ventilation.
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Objective:To develop the early respiratory rehabilitation techniques in pediatric intensive care unit(PICU) for children with mechanical ventilation.Methods:A respiratory rehabilitation team was established, and early respiratory rehabilitation intervention methods for critically ill children were also developed.Early bedside rehabilitation were implemented after the vital signs of mechanical ventilation children were stable for 24 hours, and the rehabilitation effect and adverse events were observed.Results:According to the patient′s condition, 23 patients in PICU received early respiratory rehabilitation after incubations, including 10 males and 13 females, aged 57.0(17.5, 91.5) months.There was no adverse event in 729 rehabilitation practices.The average consulting time was 8.0(4.0, 18.0) days after incubations.The average incubation time was 693.5(345.2, 1 027.1)hours of all the patients.As to 16 patients who were weaned before charged out, the average incubation time was 566.2(243.4, 928.7)hours, and the average incubation time from respiratory rehabilitation performance to weaning was 168.0(6.0, 564.0) hours.In 19 patients who only received limbs massage and passive movements, the average incubation time was 381.7(309.6, 541.4) hours and the average incubation time from respiratory rehabilitation performance to weaning was 187.2(81.5, 382.9) hours.However the difference between these who received respiratory rehabilitation and only received limbs massage and passive movements was not statistically significant.Conclusion:It′s safe and feasible to perform the early respiratory rehabilitation in PICU for critically ill children with mechanical ventilation.
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Survivors from pediatric intensive care unit are with high risk of physical, neurocognitive and psychosocial sequelae as results of critical illness.Prolonged immobility is associated with significant short- and long-term morbidities in critically ill children.The present practice on early rehabilitation, especially early mobilization, was briefly summarized in this paper.The definition, content, safety and feasibility, safe criteria, goal setting standard, as well as protocol bundles of early mobilization were all included.The safety and feasibility of early rehabilitation have been demonstrated.Stratified activity goals could be set according to the assessing results.Bringing the early mobilization into ABCDEFGH bundles may improve the recovery and outcomes for critically ill children.Possible effective ways to promote the implementation of early rehabilitation include establishing the practice guidelines, improving the physicians′ awareness of the importance and lightening their concerns on the safety of early rehabilitation, and also involving the families.
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Objective ToexploretherelationshipbetweenSchizasgradeofthenerverootwithintheduralsacandtheduralsac cross-sectionalarea(DSCA)ofthelumbarspineaswellastheclinicalsignificance.Methods 3.0T MRIexaminationofthelumbar spineof89patientswithlunbarspinestenosis(LSS)from May2016toSeptember2017intheaffiliatedhospitalofNantongUniversitywere collected.Twoexperienceddoctorsindependently measuredthekyphosisdegreeofthethoracolumbarspine,theDSCAofthe2-5 lumbarlevels,vDSCA,dDSCA,andevaluatedSchizasgradeofthenerverootforfourdegradsofA,B(gradeB1:DCSA≥100 mm2, gradeB2 :DCSA<100 mm2 ),CandDaccordingtozygopophysisconnectingline,andfinallyconductedthetestof Kappa consistency.DSCA wasdividedintothreegroupsof≤75 mm2,76-99 mm2and≥100 mm2,andχ2 wasadoptedtoexaminetherateineachSchizas grade.Schizasgradewithd/vvalue(dDSCA/vDSCA)andthekyphosisdegreeofthethoracolumbarspinewerecomparatedbyttest. Forthecorrelationcoefficient,S pear m an analysis wasadopted.Results In89cases with173lumbarlevels,schizasgradeofthenerve rootwere52,51,32and38levelsforgradeA-DrespectivelyI.nDSCA≤75mm2group,SchizasCandDwere18.5%and21.9%respectively, whichweresignificantlyhigherthanthoseforgradeAandB(0% and3.5%,P<0.01);InDSCA=76-99mm2group,Schizasgrade AandBwere8.7% and17.9%,whichweresignificantlyhigherthanthoseofgradeCandD (0% and0%,P<0.05and0.01);In DSCA≥100mm2group,therewere0% and0%forSchizasgradeCandD,whichweresignificantlylowerthanthoseforgradeAand B(21.4% and8.1%,P<0.0SchizasgradesofA-Dgroups,d/vaveragevalueswere0.64±0.29,0.48±0.22,0.42±0.20and0.34±0.11 respectively,in whichgradeCand D weresignificantlylower thanthoseofgradeAandB(P<0.01).Thecorrelationcoefficientof SchizasgradewiththeDSCAandd/vvalueswere0.83and0.87 respectively(P<0.01).Thekyphosisdegreeofthethoracolumbar spinewas(158.7±15.9)°inSchizasgradeB1,and (167.8±11.2)°inothergrades(t=4.37,P<0.05).Conclusion Theclassification ofnerverootSchizasgradeishighlyrelatedtoDCSA,andbothofthemaretheindicatorsforjudgingwhetherthelumbarspinalis stenosisornormal.TheSchizasgradeismoreconvenientandquicker;InordertoavoidconflictwithDCSA,SchizasBshouldbedividedintoB1 andB2 Whenitisusedtodeterminewhetherhavestenosis.
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Objective To observe the effects of Wenyang Baidu Yin on the serum liver DeRitis ratio [aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio] and the level of renal neutrophil gelatinase-associated lipocalin (NGAL) in septic rats. Methods One hundred and twenty healthy male sprague-dawley (SD) rats were divided into normal control group, model group, western medicine treatment group, traditional Chinese medicine (TCM) treatment group and combined medicine group by random number table method, with 24 rats in each group. Rat sepsis model was established by intra-peritoneal injection of E.coli endotoxin; after modeling for 2 hours, normal control group and model group were given normal saline intragastric administration, western medicine treatment group was given dexamethasone 2.5 mg/kg intra-gastric administration, TCM treatment group was given Wenyang Baidu Yin intragastric administration (honeysuckle 30 g, dandelion 30 g, aconite 30 g, mixed with water and after two times of boiling, the obtained liquid was concentrated and made into dry ointment for reserve, when the ointment was used, it was dissolved in warm water to 8 g/kg, administered by gavage, twice a day), the combined medicine group was given both the above TCM decoction and dexamethasone dosages by gavage. After modeling for 24, 48 and 72 hours, the differences of DeRitis ratio and NGAL levels were compared among various groups. Results The DeRitis ratio and NGAL level in the model group were significantly higher than those in the normal control group at 24, 48, 72 hours after modeling (DeRitis ratio: 4.14±0.53 vs. 2.08±0.37, 4.78±0.07 vs. 1.73±0.61, 4.83±0.31 vs. 1.05±0.20 respectively, NGAL was 2 412.73±368.03 vs. 837.52±142.98, 1 735.16±290.76 vs. 817.38±102.08, 1 838.43±295.44 vs. 847.82±140.14 respectively, all P < 0.05). The DeRitis ratios and NGAL levels of western medicine treatment group, TCM treatment group and combined medicine group were significantly lower than those of model group at each time point, and the decrease degree of combined medicine group was more significant, there were statistical differences 48 hours after modeling between combined medicine group and Western medicine treatment group and between combined group and TCM treatment group [DeRitis ratio: 3.09±0.15 vs. 3.68±0.44, 3.27±0.62, NGAL (ng/L):1 105.32±270.27 vs. 1 206.35±143.82, 1 526.53±289.02], and reached the lowest level until 72 hours [DeRitis ratio: 2.27±0.26 vs. 2.93±0.38, 2.61±0.54, NGAL (ng/L): 903.47±105.37 vs. 9 316.05±116.23, 1 025.14±152.68, all P < 0.05], showing the usage of combined therapeutic method in the aspect of improving DeRitis ratio and NGAL of septic rats was prominently superior than using either western medicine or TCM decoction alone. Conclusion Wenyang Baidu Yin can effectively reduce the liver DeRitis ratio and the kidney NGAL level in endotoxin-induced septic rats.
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OBJECTIVE: To explore the current situation of work stress,mental resilience and mental health in the local policemen,and to explore the association among these three variables. METHODS: A total of 260 local policemen in 3 districts of a city in Hebei Province were selected as the study subjects using cluster sampling method. The Chinese version of Perceived Stress Scale,Connor-Davidson Resilience Scale and General Health Questionnaire were used to evaluate their work stress,mental resilience and mental health. RESULTS: There was a negative correlation between work stress and mental resilience( P < 0. 01). Positive correlation was found between work stress and mental health( P < 0. 01). The mental resilience was negatively correlated with mental health( P < 0. 01). The mental resilience could negatively predict mental health( β =-0. 31,P < 0. 01). Work stress could positively predict mental health( β = 0. 56,P < 0. 01). Work stress had a negative prediction on mental resilience( β =-0. 47,P < 0. 01). Mental resilience played a partial mediating role between work stress and mental health( β = 0. 41,P < 0. 01). The mediating effect accounted for 26. 0% of the total effect. CONCLUSION: Mental resilience plays a partial mediating role in influencing work stress on mental health. Reducing work stress and increasing resilience play an important protective role in mental health of local policemen.
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Background Xenotransplantation is arousing more attention of researchers because of the lack of corneal donors.Biological corneal scaffolds constructed by porcine corneal acellular stroma appears to have an acceptable biocompatibility.However,its clinical effects and the histomorphological features in the corneal tissue of receipts' are still unclear.Objective This study was to evaluate the viability of bioengineered corneas as a new material of human lamellar keratoplasty and observe the in viva biological features after human keratoplasty under the laser confocal microscope.Methods A prospective serial cases observational study was carried out.Fifteen eyes of 15 patients with infectious keratitis were enrolled in Henan Eye Hospital from February to August 2014 under the approval of Ethic Committee of Henan Eye Hospital and informed consent of each patient, including 4 eyes of fungal keratitis,1 eye of bacterial keratitis, 9 eyes of mixed infectious keratitis and 1 eye of leucoma.Corneal lamellar transplantation was performed on the eyes with the bioengineered corneas as grafts and the follow-up time was one year.The survival of grafts was assessed and scored, and the indices including corneal transparency,neovascularization, graft lysis or inflammatory recurrence and visual acuity (logMAR) were evaluated.The morphology and density of corneal epithelial cells and endothelial cells, corneal stroma and subepithelial neural fibers were examined by laser confocal microscope 3,6,9 and 12 months after surgery.Results Postoperative inflammatory response was seen 3 days and disappeared 7 days after surgery.The grafts were clear 1 month after surgery, and no corneal dissolution was found during the follow-up duration.Glaucoma occurred in 1 eye at 6 months and graft rejection occurred in another eye 12 months after surgery.The logMAR, corneal transparency scores and corneal neovascularization scores improved after surgery in comparison with before surgery,with a significant difference among various time points (x2 =92.63,59.37,10.50 ,all at P<0.05).Complete epithelization of grafts was seen.Compared with the contralateral eyes,the morphology of epithelial cells was similar and endothelial cells were enlarged in the operated eyes.In addition,no stromal cell structure was seen in the corneal stroma in the operative eyes.Subepithelial nerve fibers appeared in 8 eyes at 6 months, but the fiber density was lower in the operated eyes than that in the contralateral eyes.Significant difference was found in epithelial cell density among different time points (F=1.48, P =0.22).The endothelial cell densities were (2 542 ± 119), (1 895 ± 129), (1 869 ± 135), (1 854 ± 101) and (1 844 ± 103)/mm2 before surgery and 3, 6, 9 and 12 months after surgery, showing significant differences between preoperation and postoperative time points (all at P<0.05).The subepithelial nerve fibers densities were (1.26± 0.13),(3.62±0.81) and (5.98±0.44) mm/mm2 at 6,9 and 12 months after surgery,with significant differences between adjacent two time points (t'=-8.16 ,-7.24;both at P=0.00).Conclusions Bioengineered corneal grafts survive well in human eye after lamellar transplantation,which can reconstruct ocular surface and improve the visual acuity.Biological cornea can replace human corneal materials.
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Objective To determine the reliability of Chinese-version of Cerebral Palsy Quality of Life Questionnaire for Children (CPQOL) between self-raters. Methods Family members of 21 children with cerebral palsy from rehabilitation centers or hospitals, 2 mem-bers of the same one, and 18 teenagers with cerebral palsy from a special school along with one of their parents, answered CPQOL. The in-traclass correlation coefficient (ICC) between family members, as well as teenagers and their parents were calculated, and the differences of scores of each dimension between parents and teenagers were compared. Results The reliability between family members was acceptable (ICC=0.49-0.86) in all the dimension of CPQOL, which was good in the function and acquired service dimensions, and was moderate in par-ticipation and body health dimensions. The reliability was weaker between parents and teenagers (ICC=0.29-0.73), which was acceptable in the dimensions of function, and participation and body health;and very low in the dimension of welfare and acceptance. The teenagers often rated better than their parents did, especially in the dimension of participation and body health (P<0.05). Conclusion The reliability of CPQOL is acceptable when rated by family members of cerebral palsy children, especially in the dimensions of function and service ac-quired. It is weaker between parents and children, especially in the dimension of welfare and acceptance.
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To study chemical constituents contained in ethanol extracts from roots of Machilus yaoshansis. Fifteen compounds were separated from the roots of M. yaoshansis by using various chromatographic techniques. Their structures were identified on the basis of their physicochemical properties and spectral data as twelve lignans(+)-guaiacin (1), kadsuralignan C (2), (+)-isolariciresinol (3), 5'-methoxy-(+)-isolariciresinol (4), (7'S, 8R, 8'R)-lyoniresinol (5), meso-secoisolariciresinol (6), isolariciresinol-9'-O-beta-D-xylopyranoside (7), 5'-methoxy-isolariciresinol-9'-O-beta-D-xylopyranoside (8), lyoniresinol-9'-O-beta-D-xylopyranoside (9), (2R, 3R) -2, 3-dihydro-2-(4-hydroxy-3-methoxyphenyl)-7-methoxy-3-methyl-5-(E)-propenylbenzofuran (10), 3, 5'-dimethoxy-4', 7-epoxy-8, 3'-neolignan-4, 9, 9'-triol (11), nectandrin B (12), and three flavanes(+)-catechin (13), (-)-epicatechin (14), and bis-8, 8'-catechinylmethane (15). All of the compounds 1-15 were separated from M. yaoshansis for the first time.
Subject(s)
Butylene Glycols , Chemistry , Catechin , Chemistry , Lauraceae , Chemistry , Lignans , Chemistry , Lignin , Chemistry , Naphthols , Chemistry , Plant Roots , ChemistryABSTRACT
Twenty-one non-anthraquinones constituents were isolated for the first time from an ethanol extract of the roots of Knoxia valerianoides by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were identified by their physical-chemical properties and spectroscopic analysis including NMR and MS. The compounds include ten triterpenoids: ursolic acid (1), oleanolic acid (2), 2-oxo pomolic acid (3), pomolic acid (4), maslinic acid (5), rotungenic acid (6), tormentic accid (7), rotundic acid 3,23-acetonide (8), arjungenin (9), and 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (10), four sitosterones: (24R)-24-ethylcholesta-4,22-dien-3-one (11), 3-oxo-4-en-sitosterone (12), 7-oxostigmasterol (13), and 7-oxo-beta-sitosterol (14), two lignans: eudesmin (15) and ciwujiatone (16), one coumarin: cnidilin (17), and four simple aromatic analogues: 5-hydroxymethylenefural (18), 3-hydroxy-4-methoxybenzoic acid (19), benzoic acid (20), and 2-hydroxy-5-methxoycinnamaldehydes (21). In the in vitro assays against human cancer cell lines (HCT-8, Bel7402, BGC-823, A549, and A2780), against deserum and glutamate induced PC12-syn cell damage, and against HIV-1 replication, and inhibiting protein tyrosine phosphatase 1B (PTP1 B), LPS induced NO production in macrophage, and Fe(2+)-cystine induced rat liver microsomal lipid peroxidation, at a concentration of 1 x 10(-5) mol x L(-1), no compound showed activity.
Subject(s)
Animals , Humans , Mice , Cell Line, Tumor , Chromatography, High Pressure Liquid , Lignans , Chemistry , Plant Roots , Chemistry , Rubiaceae , Chemistry , Sitosterols , Chemistry , Pharmacology , Triterpenes , Chemistry , PharmacologyABSTRACT
Two new compounds (1 and 2), together with twenty-one known compounds (3-23), were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, MCI gel, silica gel, and Sephadex LH-20 and reversed-phase HPLC. Their structures were identified by spectroscopic data analysis as 4-hydroxy-3-(4-hydroxybenzyl) benzyl methyl ether (1), 4-( methoxymethyl) phenyl-1-O-beta-D-glucopyranoside (2), hibicutaiwanin (3), 4-(4-hydroxybenzyl)-2-methoxyphenol (4), 4,4'-methylenebis(2-methoxyphenol) (5), L-phenyllactic acid (6) ,4-hydroxy-3-methoxybenzyl ethol ether (7), p-hydroxylbenzyl alcohol (8), p-hydroxylbenzyl methyl ether (9), p-hydroxylbenzyl ethyl ether (10), p-hydroxybenzaldehyde (11), p-hydroxybenzoic acid (12), p-hydroxybenzoic acid (13), gastrodin (14), 4-(ethoxymethyl) phenyl-1-O-beta-D-glucopyranoside (15), 4-(beta-D-glucopyranosyloxy) benzaldehyde (16), p-methylphenyl-1-O-beta-D-glucopyranoside (17 ), methyl-O-beta-D-glucopyranoside (18), 5-hydroxymethl-furan aldehyde (19), parishin (20), parishin B (21), parishin C (22), and diosgenin (23). The 13C-NMR data of compound 4 was first reported.
Subject(s)
Drugs, Chinese Herbal , Chemistry , Gastrodia , Chemistry , Organic Chemicals , Water , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents of the roots of Knoxia valerianoides and their biological activities.</p><p><b>METHOD</b>The anthraquinones were isolated by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Structures of the isolates were identified by their physical-chemical properties and spectroscopic analysis including 2D NMR and MS. Antioxidant, anti-HIV, neuroprotective, and cytotoxic activities were screened by using cell-based models.</p><p><b>RESULT</b>Twenty-two constituents were isolated from an ethanolic extract of the roots of K. valerianoides. Their structures were identified as nordamnacanthal (1), ibericin (2), rubiadin (3), damnacanthol (4), 2-ethoxymethylknoxiavaledin (5), 3-hydroxymorindone (6), knoxiadin (7), 2-formyl knoxiavaledin (8), lucidin (9), xanthopurpurin (10), 1, 3-dihydroxy-2-methoxy-9, 10- anthraquinone (11), lucidin(-methyl ether (12), digiferruginol (13), 3-hydroxy-2-methyl-9,10-anthraquinone (14), rubiadin-1-methyl ether (15), 6-methoxylucidin (-ethyl ether (16), 1,3,6-trihydroxy-2-methyl-9,10-anthraquinone (17), 1,3-dihydroxy-2-hydroxy methyl-6-methoxy-9,10-anthraquinone (18), 1,3,6-trihydroxy-2-methoxymethyl-9,10- anthraquinone (19), 3,6-dihydroxy-2- hydroxymethyl-9,10-anthraquinone (20), and 1,6-dihydroxy-2-methyl-9,10-anthra quinone (21). In the in vitro assays, at a concentration of 1 x 10(-5) mol x L(-1), no compounds were active against human cancer cell lines (HCT-8, Bel7402, BGC-823, A549, and A2780), deserum and glutamate induced PC12-syn cell damage, LPS induced NO production in macrophage, Fe2+-cystine induced rat liver microsomal lipid peroxidation, HIV-1 replication, and protein tyrosine phosphatase 1B (PTP1B).</p><p><b>CONCLUSION</b>Compounds 9-21 were obtained from the roots of K. valerianoides for the first time.</p>
Subject(s)
Animals , Humans , Rats , Anthraquinones , Chemistry , Pharmacology , Cell Line , Drugs, Chinese Herbal , Chemistry , Pharmacology , Molecular Structure , Plant Roots , Chemistry , Rubiaceae , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate chemical constituents of the stems and branches of Adina polycephala and their pharmacological activities.</p><p><b>METHOD</b>The constituents were isolated by a combination of various chromatographic techniques including column chromatography on silica gel, Sephadex LH-20, and C-18, as well as reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis. In vitro cytotoxic, anti-inflammatory, anti-oxidant, anti-HIV, neuroprotective and anti-diabetic activities were screened by using cell-based models.</p><p><b>RESULT</b>Twenty-eight constituents were isolated. Their structures were identified as clemochinenoside B (1), kelampayoside A (2), osmanthuside H (3), 4-hydroxy-3-methoxyphenol-beta-D-[6-O-(4-hydroxy-3,5-dimethoxylbenzoate)]-glucopyranoside (4), and syringic acid beta-D-glucopyranosyl ester (5). Ten iridoidal glycosides: geniposidic acid (6), geniposide (7), 6beta-hydroxygeniposide (8), 6beta-hydroxygeniposide (9), ixoside (10), ixoside 11-methyl ester (11), 11-methyl forsythide (12), 7beta-hydroxysplendoside (13), gardoside (14) and mussaenosidic acid (15), (+) -pinoresinol (16), (+) -medioresinol (17), (+) -syringaresinol (18), (-)-lariciresinol (19), evofolin-B (20), alpha-hydroxyacetovaillone (21), syringic acid (22), vanillin (23), 3, 4, 5-trimethoxyphenol (24), and 2,6-dimethoxy-1, 4-benzoquinone (25), beta-sitosterol (26), mannitol (27), and daucosterol (28). At a concentration of 1.0 x 10(-5) mol x L(-1), these compounds were inactive in the assays, including cytotoxicity against human tumor cell lines (HCT-8, Bel-7402, BGC-823, A549 and A2780), anti-inflammatory activity against the release of beta-glucuronidase in rat polymorphonuclear leukocytes (PMNs) induced by platelet-activating factor (PAF), antioxidant activity in Fe(2+)-cystine-induced rat liver microsomal lipid peroxidation, anti-HIV activity against HIV-1 replication, neuroprotective activity against serum deprivation or glutamate induced neurotoxicity in cultures of PC12 cells, and the inhibitory activity against protein tyrosine phosphatase 1B (PTP1B).</p><p><b>CONCLUSION</b>Compounds 1-20 were obtained from the genus Adina for the first time. The 13C-NMR data of compounds 10 and 11 were reassigned. A further evaluation of pharmacological activity of these compounds is expected.</p>