BACKGROUND:As a degradable scaffold material for bone tissue engineering,lactic acid is widely used in tissue regeneration and repair research,and plays an important role in promoting tissue healing,new bone formation and angiogenesis. OBJECTIVE:To observe the effect of lactic acid degradation products on osteoclasts and to investigate the effects of lactic-interfered osteoclast conditioned medium on the proliferation,migration and tube-forming capacity of human umbilical vein endothelial cells. METHODS:(1)The mouse monocyte macrophage cell line RAW264.7 at logarithmic growth period was selected,and adherent cells were cultured in the osteoclast induction medium(DMEM medium with nuclear factor-κB receptor-activating factor ligand and 10%fetal bovine serum)containing different concentrations of lactic acid(0,5,10,20 mmol/L).After 5 days of culture,tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining were conducted.After 24 hours of culture,RT-PCR was used to detect the mRNA expression of tartrate-resistant acid phosphatase 5.(2)RAW264.7 cells at logarithmic growth period were selected and adherent cells were divided into two groups.Control group was cultured in the osteoclast induction medium,while experimental group was cultured in the osteoclast induction medium containing 10 mmol/L lactic acid.After 5 days of culture,the medium in each group was removed and the cells in the two groups were cultured in the serum-free DMEM medium for another 24 hours.Cell supernatant was then collected and used as the conditioned medium after mixed with an equal volume of DMEM medium containing 10%fetal bovine serum.Human umbilical vein endothelial cells at the logarithmic growth phase were taken and separately co-cultured with the conditioned medium of the control and experimental groups.The proliferation,migration and tube-forming ability of human umbilical vein endothelial cells were observed by cell counting kit-8 assay,migration assay,scratch assay and tube-forming assay.The mRNA and protein expression of angiogenesis-related genes and proteins were observed by RT-PCR and western blot. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining showed that 5 and 10 mmol/L lactic acid promoted osteoclastic differentiation of RAW264.7 cells and the promoting effect of 10 mmol/L lactate was more significant.RT-PCR results showed that the expression of tartrate-resistant acid phosphatase-5 mRNA of osteoclast-related genes was the highest when the lactic acid concentration was 5,10,and 20 mmol/L(P<0.05),especially 10 mmol/L.Compared with the control group,the proliferation,migration and tube-forming abilities of human umbilical vein endothelial cells were significantly increased in the experimental group(P<0.05).Compared with the control group,the expression levels of vascular endothelial growth factor and angiogenin 1 mRNA and protein were increased in the experimental group(P<0.05).To conclude,lactate-induced osteoclast conditioned medium could promote the angiogenesis of endothelial cells,and the mechanism may be related to the promotion of the expression of vascular endothelial growth factor and angiogenin 1.

BACKGROUND:Three-dimensional(3D)printing is an emerging technology in the field of dentistry.It utilizes a layer-by-layer manufacturing technique to create scaffolds suitable for periodontal tissue engineering applications.Tissue scaffolds produced through 3D printing can possess controlled characteristics,including internal structure,porosity,and interconnectivity,making it an ideal strategy for periodontal tissue engineering. OBJECTIVE:To review the applications of 3D printed scaffolds in periodontal regeneration. METHODS:English search terms were"3D printing,periodontal tissue engineering,additive manufacturing,regenerative medicine,bioengineering,scaffold,bioprinting,periodontitis".Chinese search terms were"3D printing,additive manufacturing,periodontal tissue engineering,scaffolds,bio-inks,bioprinting,tissue engineering".Relevant literature published from 2000 to 2023 in PubMed and CNKI databases was retrieved and included in the review. RESULTS AND CONCLUSION:Over the past few decades,3D printing technology has made significant progress and breakthroughs in tissue engineering and biomedical fields.3D printing technology can provide highly personalized treatment programs,improve the suitability and therapeutic effect of therapeutic stents,and has broad application prospects in periodontal tissue engineering.In periodontal tissue engineering,3D printing applications can better mimic the complex structures of biological tissues and manufacture biocompatible scaffold materials with suitable mechanical and rheological properties.The layer-by-layer construction of tissue engineering scaffolds through 3D printing not only enables the creation of precise and intricate scaffold models for personalized treatment of periodontal disease but also facilitates the incorporation of complex microstructures and channels within the scaffolds to promote cell growth and tissue regeneration.

Purpose To examine the clinicopathologic,immunohistochemical,and molecular genetic characteristics and differential diagnosis of low-grade oncocytic tumor(LOT)of kidney with CK7 positive and CD117 negative,so as to enhance the understanding of this tumor among pathologists.Methods A total of five cases of renal LOT from the First Affiliated Hospi-tal of Soochow University between December 2016 and February 2023 were included in this analysis.The clinicopathological fea-tures,immunophenotype,genetic characteristics,and prognosis were evaluated using HE staing,immunohistochemical staining,and Sanger sequencing.Additionally,relevant literature was re-viewed to supplement the findings.Results Among the cohort of five patients,four were female and one was male,aged 57-70 years with a median age of 65 years and an average age of 64.8 years.Clinical presentation revealed that only the first case exhibited frequent urination accompanied by intermittent lumba-go,while the remaining cases were asymptomatic and incidental-ly discovered.Imaging studies demonstrated space-occupying le-sions with clear boundaries and even internal echoes on B-ultra-sonography,and patchy low-density shadows in the center with obvious edge enhancement on CT.Grossly,the tumor was nodu-lar,with a maximum diameter ranging from 2.1 to 7.6 cm,and an average diameter of 4.04 cm,with a solid section.Micro-scopically,the boundary of LOT was consistently well-defined,with a thick capsule.The arrangement of tumor cells was ob-served to be both dense and sparse,accompanied by fresh bleed-ing foci and proteinoid secretions.Focal lymphocyte aggrega-tion,hepatic plate like and hepatic sinusoid like structures,thick-walled blood vessels,false nodules of tumor cells,and old hemorrhage were also noted.The tumor cells exhibited uniformi-ty in shape,appearing round or polygonal,with eosinophilic and fine granular cytoplasm.The nuclei were of similar size and shape,appearing round or oval with a clear nuclear membrane.The histological features of the tumor included 2-grade small nu-cleoli,perinuclear halos,binuclear cells,and nuclear shrink-age,but no mitotic figures were detected.The immunophenotyp-ic analysis revealed strongly diffuse expression of CK7 in the tumor cells,while CD117 was negative.The Ki67 proliferation index was low.Sanger sequencing identified mutations in mTORC1 pathway genes in four cases,including three mutations in MTOR and one mutation in RHEB.The patients underwent either local or radical nephrectomy,and were followed up for a period ranging from 2 to 52 months,during which all patients re-mained free of recurrence.Conclusion The low-grade,eosino-philic,and rare renal tumor known as LOT exhibits inert behav-ior.At present,the follow-up results show that complete resec-tion of local operation is sufficient,and the prognosis is good.It is important to distinguish this lesion from other eosinophilic re-nal tumors.

Objective To explore the possible mechanisms of myocardial injury induced by simulated weightlessness in space flight and the recovery of myocardium in rats by aerobic exercise intervention.Methods The body weight of rats was weighed using an electronic balance;the cardiac function indexes of rats in each group were detected using echocardiography and isolated cardiac perfusion;and the expression of proteins related to the AMPK/ULK1/Beclin1 pathway in the myocardial tissues of rats in each group was detected using Western Blot.Results(1)When compared with the C1 group,the T1 group not statistically significant in body weight(P?>?0.05),and the wet weight of the flounder muscle,the wet weight-to-weight ratio of the flounder muscle,and the heart weight were all significantly decreased(P??0.05).After 4 weeks of aerobic exercise,when compared to group C2,group R showed a significant rise in the expression of AMPK and ULK1(P??0.05),and a rise in the expression of Beclin1,but not statistically significant(P?>?0.05);and group A showed a slight increase in the expression of AMPK,ULK1 and Beclin1 expression decreased slightly but not statistically significant(P?>?0.05),p-AMPK expression decreased significantly(P??0.05),and a significant decrease in the expression of ULK1(P??0.05);the expression of p-ULK1 decreased significantly(P?

Objective To investigate the significance of auditory brainstem response(ABR)combined with auditory steady-state response(ASSR)for the assessment of mild sensorineural hearing loss in infants.Methods Data from 114 infants with mild sensorineural hearing loss were retrospectively analyzed,and their ABR and ASSR results were collected for rank sum test and correlation analysis.Results In the rank sum test,the difference in thresholds between tone-burst ABR(Tb-ABR)and ASSR at 0.5,1,2,4 kHz was statistically significant(P＜0.05),and they were also correlated at 0.5,1,2,4 kHz(P＜0.05),r=0.613,0.569,0.616,0.71.After grouping by gender and ear,there was a correlation between ABR and ASSR at 0.5,1,2,and 4 kHz,male:r=0.61,0.56,0.671,0.774;female:r=0.581,0.558,0.546,0.608;left ear:r=0.61,0.558,0.576,0.715;right ear:r=0.631,0.581,0.662,0.71.And after grouping by age at diagnosis,only infants diagnosed from 7～12 months of age did not correlate at 0.5 kHz and 1 kHz(P＞0.05),while the rest of the groups had a good correlation(P＜0.05),0～3 months:r=0.686,0.643,0.671,0.742;4～6 months:r=0.671,0.626,0.616,0.693;7～12 months at 2 kHz and 4 kHz:r=0.571,0.706.Conclusion In infants with mild sensorineural hearing loss,ABR and ASSR correlate in assessing hearing thresholds at all frequencies,and the combination of the two tests could provide a more accurate assessment of the subject's true hearing.

Humans ; Proto-Oncogene Proteins p21(ras)/genetics* ; Prognosis ; Biomarkers, Tumor/genetics* ; Mutation/genetics* ; Colorectal Neoplasms/genetics* ; Proto-Oncogene Proteins B-raf/metabolism*

Objective To investigate the role and mechanism of SR8278,a synthetic antagonist of nuclear receptor subfamily 1 group D member 1(NR1D1),in alleviating the structural and functional impairment of the extraorbital lacrimal glands induced by jet lag in mice.Methods Totally 36 healthy wild C57BL/6J mice aged 8-10 weeks were randomly divid-ed into 3 groups(normal group,jet-lag group,and jet-lag+SR8278 group)after adapting to a circadian rhythm chamber under the 12 h light/12 h dark(12 h/12 h LD)cycle for 2 weeks,with 12 mice in each group.Mice in the normal group were fed in a circadian rhythm chamber in a 12 h LD cycle,mice in the jet-lag group were fed in a 12 h/12 h LD cycle with an 8-hour advanced LD schedule,and mice in the jet lag+SR8278 group were fed in a 12 h/12 h LD cycle with an 8-hour advanced LD schedule and received 25 mg·kg-1 SR8278.At the end of 5 days of intervention,locomotor activity,core body temperature and tear secretion of mice in each group were collected,and the weight of lacrimal gland tissues and size of lacrimal gland cells were measured.Immunohistochemical methods were used for histological evaluation of the extraor-bital lacrimal glands in mice.Lacrimal ribonucleic acid(RNA)was extracted for high-throughput RNA-sequencing analysis containing NR1D1,and the obtained transcriptomic data were used for KEGG and GO functional enrichment analysis.Re-sults Compared with the normal group,the jet-lag group had higher daytime activity,lower nighttime activity,higher daytime core body temperature,and lower nighttime core body temperature,with statistically significant differences(all P＜0.05).Compared with the jet-lag group,the jet-lag+SR8278 group had lower daytime activity,higher nighttime activi-ty,lower daytime core body temperature,and higher nighttime core body temperature,with statistically significant differ-ences(all P＜0.05).Compared with the normal group,the jet-lag group showed a decrease in lacrimal gland weight and tear secretion and an increase in size of lacrimal gland cells,with statistical significance(all P＜0.05);compared with the jet-lag group,the jet-lag+SR8278 group had an increase in lacrimal gland weight and tear secretion and a decrease in size of lacrimal gland cells,with statistical significance(all P＜0.05).Compared with the normal group,the jet-lag group showed a higher expression of NR1D1 in the lacrimal gland at night;compared with the jet-lag group,the jet-lag+SR8278 group showed a lower expression of NR1 D1 in the lacrimal gland at night(both P＜0.05).Bioinformatics analysis showed 947 significantly different genes in the jet-lag group and the jet-lag+SR8278 group,of which 43 are significantly upregulated genes,and 904 are significantly downregulated genes.The Notch signaling pathway has the most significant difference.Conclusion SR8278 effectively enhances the tear secretion function of jet-lagged mice by targeting NR1D1 inhibition.This process may be completed through the Notch signaling pathway.

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.