BACKGROUND:Osteoblasts with high purity and activity are essential for bone metabolism research. OBJECTIVE:To explore a simple and effective culturing method of primary osteoblasts. METHODS:Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope;the cells was counted to draw the growth curve;the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining. RESULTS AND CONCLUSION:The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day, and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts.

Objective To investigate the methods of nursing safety management for indwelling catheters. Methods The nursing safety management measures included the establishment of three-level monitoring network of safety management for indwelling catheters,standardization of 25 kinds of catheter identification cards,formulation of prevention procedure and reporting system of catheter extrusion,design of catheter management handbook,organization of training and examination on the relative skills for nurses. Results Nurses' safety awareness of indwelling catheters was strengthened. And nurses could distinguish different catheters promptly and provide efficacious and safe nursing services. There was no nursing error,nursing dispute and patient complaints towards nursing care of indwelling catheters. Conclusion The implementation of nursing safety management for indwelling catheters can ensure medical and nursing safety,and promote the continuous improvement of quality of nursing.

Objective:To summarize the nursing care of a super old patient with right limb movement disorder accompanied by mixed aphasia and cardiac and renal insufficiency who had placed PICC catheter on the same side of pacemaker for multiple ectopic adjustments.Method:Full evaluation before catheterization and comprehensive nursing after catheterizationResult:No complications such as catheter blockage, thrombus, swelling and infection were found in the patients who were treated with PICC catheter for 80days.Conclusion:Before placing PICC catheter on the same side of the pacemaker, comprehensive evaluation and targeted nursing should be given to ensure the safe retention of PICC and the normal operation of the pacemaker.

To summarize the nursing experience of a child with severe cardiopulmonary failure caused by fulminant myocarditis who underwent autologous vascular bypass grafting after severe lower limb ischemia during extracorporeal membrane oxygenation therapy.The key points of nursing include:①keeping the distal perfusion tube unobstructed before surgery to avoid aggravation of ischemia of the affected limb;physical precautions for deep vein thrombosis are strictly implemented to ensure the patency of the saphenous vein.②intraoperative nursing cooperation for the conversion of venous-arterial-venous extracorporeal membrane oxygenation to venous-venous extracorporeal membrane oxygenation.③on the premise of ensuring systemic perfusion after surgery,the blood pressure should be strictly controlled to prevent the rupture of the grafted vessel;to strengthen the management of coagulation to prevent bleeding and thrombosis;different position management of both lower limbs to keep the grafted blood vessels patency;to develop individualized exercise programs to enhance nutrition and promote rapid recovery;to give individualized psychological nursing care including childlike fun and sense of security,and eliminate children's negative emotions;to develop systematic health guidance and follow-up to improve the quality of family care for children.After treatment and nursing care,the postoperative grafted blood vessels were well anastomosed with no limb ischemia,and there was no swelling of the limb in the saphenous vein donor area.After 41 days of the treatment,the extracorporeal membrane oxygenation was withdrawn.The child recovered and discharged after 122 days;after 2 years of follow-up,the child could walk normally and recovered well.

Objective To explore the influence of family-integrated transition care on the daily living ability of discharged patients with stroke.Methods Seventy-eight patients with stroke who were admitted to Renmin Hospital of Wuhan University from May 2016 to October 2017 were selected by convenience sampling and were divided into a control group and a family-integrated transition care group (hereinafter referred to as transition care group).The patients in the control group received routine neurological health education,while those in the transition care group received a family-integrated transition care intervention in addition to routine neurological health education.The family-integrated transition care included team building,skills training for family members,family-integrated guidance for discharged patients,and regular visits.The scores of the modified Barthel index were compared between the two groups of patients at discharge,three months after intervention,and six months after intervention.Results Among the 71 patients that were finally included,35 cases were included in the control group,of which 17 cases were men (49％),18 cases were women (51％),and their mean age was (70.1±3.7) years;the transition care group comprised 36 cases,of which 18 cases were men (50％),18 cases were women (50％),and their mean age was (69.8±4.5) years.The baseline scores of the control group and transition care group on the day of discharge were (49.1 ± 7.5) and (49.7 ± 7.9),respectively,with no significant difference (P＞0.05).In terms of time effects,the scores of the patients in the two groups had statistically significantly improved at six months after discharge (P＜0.05).In the group comparison,the scores of the patients in the transition care group after the intervention were significantly higher compared to the scores of those in the control group (P＜0.05).In terms of time and inter-group effects,there was an interaction (P＜0.05),and therefore,the influence of time effects was excluded and the same timepoint was compared between the two groups.The scores at three months (63.9±8.8) and six months (76.9± 10.1) in the transition care group were higher than those in the control group (58.1 ±8.1 and 66.0 ±9.3,respectively).The difference was statistically significant (P ＜ 0.05).Conclusion Family-integrated transition care can effectively improve daily living ability and isworthy of promoting.

Objective:To explore the effect of pelvic floor muscle functional exercise based on Snyder hope theory in patients after prophylactic stoma retraction.Methods:74 patients with low rectal cancer who underwent prophylactic stoma retraction from July 2019 to June 2021 were randomly divided into intervention group and control group. The patients in the control group received routine nursing and pelvic floor muscle functional exercise. The patients in the intervention group received functional exercise intervention based on Snyder′s hope theory on the basis of the control group. The hope level and self-care ability of the patients in the two groups were evaluated before the intervention and 3 months after stoma restitution. The anal function of the patients in the two groups was evaluated 1 month and 3 months after stoma restitution.Results:Before the intervention, there was no significant difference in the score of hope level and self-care ability between the two groups ( P>0.05). Three months after the operation, the score of hope level in the observation group was 36.20 ± 3.82, which was higher than that in the control group (31.26 ± 5.03) ( t = 4.63, P<0.05). Three months after the operation, the self-care ability score of the observation group was 123.57 ± 10.82, which was higher than that of the control group (108.23 ± 9.48) ( t = 6.31, P<0.05). One month and three months after stoma retraction, the anal function scores of the observation group were 12.03 ± 3.94, 5.91 ± 2.05 respectively, which were lower than those of the control group (13.86 ± 2.19, 7.26 ± 1.74) ( t = 2.40, 2.99, both P<0.05). Conclusion:Pelvic floor muscle functional exercise based on Snyder′s hope theory can improve the hope level of patients after stoma retraction, improve their anal function and improve their self-care ability.

This review summarized the pathogenesis of coagulation dysfunction, high-risk factors for bleeding and thrombosis, monitoring indicators for anticoagulation, drug therapy for coagulation dysfunction, assessment and nursing interventions for coagulation dysfunction in patients supported by extracorporeal membrane oxygenation, with the aim of providing evidence-based strategies for anticoagulation management in extracorporeal membrane oxygenation patients.

To investigate the effects of ORF9 gene of porcine circovirus type 2(PCV2)on PK-15,eu-karyotic expression plasmid was constructed and transfected into PK-15 cells,and the effects of overexpression of ORF9 on proliferation,apoptosis and immunization of PK-15 cells were exam-ined by flow cytometry and qRT-PCR.The results showed that ORF9 gene overexpression signifi-cantly up-regulated the expression levels of the ER stress marker gene GRP78,increased the num-ber of S phase cells,accelerated cell cycle progression,increased the apoptosis rate of PK-15 cells,up-regulated the expression levels of apoptosis-related genes caspase-3,caspase-8,caspase-9,p53 and Bax(P＜0.01),down-regulated the expression levels of apoptosis-related genes Bcl-2,up-reg-ulated the expression levels of immune-related genes 1L-8,IL-10,NF-κB and TNF-α(P＜0.01),and down-regulated the expression levels of immune-related genes IL-2,IFN-β and IL-12(P＜0.01).The above results indicate that ORF9 gene may promote the proliferation and apoptosis of PK-15 cells and play a role in the escape process of PK-15 cells.

In order to understand the genomic characteristics and genetic variation and strain type of infectious bursal disease virus(IBDV)isolate GZGY2022,which caused the death of chickens in Guizhou farm,primers were designed to amplify the whole genome of the isolate,and genetic evo-lution and strain type analysis were performed after cloning and sequencing.The results showed that the A and B segments of IBDV genome were 3 260,2 827 bp,respectively,encoding VP2-VP5 and VP1 genes.The nucleotide sequence homology between the A and B segments of this strain and the VvIBDV were 96.2％-98.7％and 87.7％-98.9％,respectively,which is the highest with NN1172 strain,83.1％-94.7％and 90.1％-91.0％with other strains.The results of genetic evolution and strain type study showed that IBDV strains can be divided into 6 branches according to antigen and virulence,and the A and B segments of the strain were clustered in the evolutionary branch of VvIBDV,and the strain was A3B3 genotype according to the new genotype classification method.The results of amino acid sequence analysis showed that there were 3 and 7 unique amino acid site variations in the A and B segments of the strain,respectively,and 13 unique characteristic amino acid sites in the coding region of the full-length genome were consistent with VvIBDV.The VP2 sequence of segment A has 19 characteristic amino acid identical with VvIBDV,among which hyper variable regions 222A,242I,253Q,256I,279D,284A,294I and 299S were characteristic ami-no acid sites of the VvIBDV,and the heptapeptide region sequence SWSASGS was consistent with the virulent strain.The VP1 sequence of segment B has 10 characteristic amino acid identical with VvIBDV,among which 61I,145T and 287A were the characteristic amino acid sites of the VvIB-DV.In addition,the nucleotide sequence GGTGCC of 777-782 did not form the restriction endo-nuclease site of Kpn Ⅰ,and combined with the triplet site 145/146/147(TEG),the segment B was consistent with the NN1172 strain,showed that its virulence was slightly weaker than that of the B2 strain of VvIBDV.The results of recombination analysis showed that there were no breaks and recombination sites in the sequence of the strain,and no recombination event occurred.In summa-ry,this study found that GZGY2022 strain belonged to the A3B3 genotype non-recombinant VvIB-DV strain,and its special amino acid sites were consistent with the molecular characteristics of VvIBDV.This study lays the foundation for further exploring the genomic characteristics and path-ogenicity of VvIBDV.

Endo-periodontal lesions(EPLs)involve both the periodontium and pulp tissue and have complicated etiologies and pathogenic mechanisms,including unique anatomical and microbiological characteristics and multiple contributing factors.This etiological complexity leads to difficulties in determining patient prognosis,posing great challenges in clinical practice.Furthermore,EPL-affected teeth require multidisciplinary therapy,including periodontal therapy,endodontic therapy and others,but there is still much debate about the appropriate timing of periodontal therapy and root canal therapy.By compiling the most recent findings on the etiology,pathogenesis,clinical characteristics,diagnosis,therapy,and prognosis of EPL-affected teeth,this consensus sought to support clinicians in making the best possible treatment decisions based on both biological and clinical evidence.