Objective:To investigate the role of ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50) in renal tubular epithelial-mesenchymal transition (EMT) in diabetic nephropathy (DN).Methods:Renal tissue specimens from 24 DN patients (DN group) confirmed by medical history, clinical laboratory tests, and pathological diagnosis at the Third Hospital of Hebei Medical University and 15 adjacent normal renal tissues (control group) from 2018 to 2023 were collected. Immunohistochemical staining was performed to detect the expression of EBP50, E-cadherin, and Vimentin proteins. Additionally, 10 healthy volunteers were enrolled as a normal group, and fasting blood glucose, serum creatinine, blood urea nitrogen, and 24 h urinary protein levels were compared between DN patients and the normal group.Results:Fasting blood glucose, serum creatinine, blood urea nitrogen, and 24 h urinary protein levels were significantly higher in the DN group than in the normal group (all P<0.01). Periodic acid-schiff (PAS) staining revealed significant thickening of the glomerular basement membrane, increased extracellular matrix, vacuolar degeneration of renal tubules, and elevated extracellular matrix in the renal interstitium in the DN group. Immunohistochemistry showed higher expression of EBP50 and E-cadherin proteins in the control group than in the DN group (all P<0.01), while Vimentin expression was lower in the control group ( P<0.01). Conclusions:EBP50 is involved in the EMT process of renal tubules in DN and is associated with tubular injury.

Objective:To examine the postoperative morphological changes in outer retinal wave-like changes (ORC) in eyes with rhegmatogenous retinal detachment (RRD).Methods:A retrospective clinical study. From March 2020 to March 2024, 64 eyes of 64 RRD patients diagnosed at The Second Affiliated Hospital of Zunyi Medical University were included. The cohort included 39 males (39 eyes) and 25 females (25 eyes), with a mean age of (45.3±15.7) years and a mean retinal detachment duration of (16.6±13.5) days. Macular involvement was observed in 51 eyes. Scleral buckling surgery (SB), pars plana vitrectomy (PPV), and SB+PPV were performed in 17, 44, and 3 eyes respectively. Best corrected visual acuity (BCVA) and spectral domain optical coherence tomography examinations were performed in all affected eyes. BCVA examination was performed using the standard logarithmic visual acuity chart, and the visual acuity was converted to logarithm of the minimum angle of resolution (logMAR) during statistics. Combined with the ORC situation before the operation, the morphological repositioning of outer retinal folds (ORF) after the operation was classified into types Ⅰ, Ⅱ and Ⅲ. One week and one and three months after the operation, relevant examinations were performed using the same equipment and methods as before the operation. The structural characteristics of ORC and the morphology of ORF after surgery were observed. The comparison between groups was conducted using the independent sample t test or the Mann-Whitney U test. Results:Among the 64 eyes, preoperative ORC was present in 46 eyes (71.9%, 46/64) and absent in 18 eyes (28.1%, 18/64). The 18 eyes without ORC had retinal detachment durations of either ≤4 days or ≥42 days. Postoperatively, ORF was observed in 51 eyes (79.7%, 51/64) and absent in 13 eyes (20.3%, 13/64). Among the 51 eyes with macular holes involved, 24 eyes (47.1%, 24/51) had ORF after the operation and 27 eyes (52.9%, 27/51) had no ORF. Among the 46 and 18 eyes with and without ORC, the ORF after surgery was 28 (60.9%, 28/46) and 1 (5.6%, 1/18) eyes, respectively. There was a statistically significant difference in the reduction rate of ORF after surgery between eyes with and without ORC ( χ2=15.974, P<0.001). Among the 46 eyes with ORC, the proportions of ORF to types Ⅰ, Ⅱ and Ⅲ after surgery were 24 (52.2%, 24/46), 20 (43.5%, 20/46), and 2 (4.3%, 2/46) eyes, respectively. There was no statistically significant difference in the incidence of ORF after different surgical methods in eyes with ORC ( P>0.05). One week and one month after the surgery, the logMAR BCVA of the affected eyes with and without ORF was 0.97±0.47, 0.69±0.34 and 0.85±0.32, 0.54±0.21, respectively. The BCVA of those without ORF was better than that of those with ORF, but the differences were not statistically significant ( t=0.237, 0.408; P>0.05). Conclusions:The occurrence of ORC in RRD eyes has a certain relationship with the time of retinal detachment. For RRD eyes with ORC before the operation, the repositioning morphology after the operation is more likely to show changes in ORF. The transformation from ORC to ORF after ORC surgery has no correlation with the surgical method.

Objective:To compare the changes of alveolar bone thickness and height in patients with bimaxillary dentoalveolar protrusion after the treatment with labial straight wire appliance and personalized lingual appliance for the retraction of anterior teeth.Methods:31 cases of labial fixed orthodontic treatment and 31 of personalized lingual orthodontic treatment were included.CBCT images were taken before and after treatment,the thickness and height of the alveolar bone on both sides of the upper anterior teeth were measured and statistically analyzed.Results:After the upper anterior teeth were retracted with labial aligner,the thick-ness of the alveolar bone in the labial root middle 1/2 and apical region of the upper incisors increased,the thickness of the alveolar bone in the cervical region of the labial side of the upper canine increased(P＜0.05),whereas the thickness and height of the alve-olar bone in the palatal side of the upper anterior teeth decreased,and the total thickness of the alveolar bone in the apical region decreased(P＜0.05).After the anterior teeth retracted with lingual aligners,the alveolar bone thickness of the labial cervical of the upper middle incisors and lateral incisors decreased,the alveolar bone thickness of the apical region of the upper middle incisors and upper canine increased,and the thickness and height of the palatal alveolar bone of the upper anterior teeth decreased,with the total thickness of the apical region decreased(P＜0.05).The reduction of alveolar bone thickness of the labial neck and root middle 1/2 of the upper incisors and alveolar bone height of the palatal side of the upper canine in the lingual appliance group was more than that in the laial group(P＜0.05).Conclusion:Labial straight wire appliance and personalized lingual appliance are effective in the retraction of the anterior teeth and the alveolar bone remodeling based on bone resorption,labial appliance is more effective.

Objective:To compare the changes of alveolar bone thickness and height in patients with bimaxillary dentoalveolar protrusion after the treatment with labial straight wire appliance and personalized lingual appliance for the retraction of anterior teeth.Methods:31 cases of labial fixed orthodontic treatment and 31 of personalized lingual orthodontic treatment were included.CBCT images were taken before and after treatment,the thickness and height of the alveolar bone on both sides of the upper anterior teeth were measured and statistically analyzed.Results:After the upper anterior teeth were retracted with labial aligner,the thick-ness of the alveolar bone in the labial root middle 1/2 and apical region of the upper incisors increased,the thickness of the alveolar bone in the cervical region of the labial side of the upper canine increased(P＜0.05),whereas the thickness and height of the alve-olar bone in the palatal side of the upper anterior teeth decreased,and the total thickness of the alveolar bone in the apical region decreased(P＜0.05).After the anterior teeth retracted with lingual aligners,the alveolar bone thickness of the labial cervical of the upper middle incisors and lateral incisors decreased,the alveolar bone thickness of the apical region of the upper middle incisors and upper canine increased,and the thickness and height of the palatal alveolar bone of the upper anterior teeth decreased,with the total thickness of the apical region decreased(P＜0.05).The reduction of alveolar bone thickness of the labial neck and root middle 1/2 of the upper incisors and alveolar bone height of the palatal side of the upper canine in the lingual appliance group was more than that in the laial group(P＜0.05).Conclusion:Labial straight wire appliance and personalized lingual appliance are effective in the retraction of the anterior teeth and the alveolar bone remodeling based on bone resorption,labial appliance is more effective.

Objective:To apply mass spectrometry as well as bioinformatics techniques to analyze HBV peptides derived from Pol and X proteins presented by human immortalized B lymphocytes(BLCLs),and to screen for effective T-cell epitopes,which lays the foundation for the development of therapeutic vaccines.Methods:The group has constructed a genome-wide expression plasmid containing 1.2-fold HBVC2 isoforms and transfected it into BLCLs by electro-transfection,isolated and identified HBV peptides by LC-MS/MS,bioinformatically predicted peptide sequences in terms of sensitization,antigenicity,toxicity,HLA molecular affinity,and the ability of peptide HLA-class Ⅰ complex to bind to specific T cells,and retrieved reported sequences.Results:HBV peptides from lysates of five immortalized B cells(BLCLs-1 to BLCLs-5)carrying HBVC2 subtype-expressing recombinants were analyzed,and 141 peptides with sequence lengths of no less than 8 amino acid residues(≥8 aa)were successfully isolated and identified,of which 133 were derived from Pol,and 8 from X proteins.The 141 HBV peptides were analyzed for sensitization,antigenicity and toxicity,and 50 antigenic,non-toxic and sensitizing HBV peptides(47 from Pol and 3 from X protein)were screened for affinity analysis with HLA-class Ⅰ and Ⅱ molecules and for prediction of the binding ability of the peptide HLA-class Ⅰ complexes to specific T cells.Among these peptides,37 had affinity to the corresponding genotypes of HLA-class Ⅰ and Ⅱ molecules.Finally,we obtained 37 peptides with antigenic,nontoxic,and sensitizing properties with IC50<500 nmol/L to HLA-class Ⅰ and Ⅱ molecules,which have affinitied to the corresponding genotypes but have not been reported,and can be continued to be explored as potential epitopes.Finally,15 HBV peptide hotspot core regions were formed by intra-and inter-strain common,contained,overlapping or neighboring sequence analysis of 141 peptides,with 7 core region sequences with antigenic,nontoxic,and sensitizing properties restricted to the corresponding geno-types.Peptides with affinity IC50<500 nmol/L had an affinity core sequence overlap of no less than 8 amino acids,and can be priori-tized for candidate T cell epitopes for subsequent studies.Conclusion:Peptides derived from Pol and X proteins have been successfully isolated and identified,and potential T cell epitopes have been screened.

Objective To study the effects and the possible mechanism of the hearing loss of offspring mice with chronic intrauterine hypoxia.Methods Ten healthy SD pregnant mice with SPF level were selected to establish an intrauterine hypoxia model as the hypoxia group,while another ten healthy SD pregnant mice without hypoxia treatment were selected as the control group.Arterial blood gas analysis was performed on pregnant mice and audi-tory brainstem response(ABR)testing was performed on their offspring after birth,and the results of each group were compared.Results The PaO2 and SaO2 of the hypoxic group were significantly lower than those of in the con-trol group.The hearing loss rate and wave Ⅲ response threshold of the offspring mice of the hypoxia group were significantly higher than those of in the control group at 21 and 56 days,but over time,the hearing loss rate and wave Ⅲ response threshold of the hypoxia group improved compared to before.Although some offspring mice in the hypoxic group at 21 day and 56 day had normal Ⅲ wave response thresholds,their wave Ⅱ and Ⅲ latency,as well as the latency between Ⅰ-Ⅱ,Ⅱ-Ⅲ,and Ⅰ-Ⅲ,were still significantly higher,while the amplitude of wave Ⅲ and the ratio of Ⅲ/Ⅰ were significantly lower than those of in the control group during the same period.For hypoxic off-spring mice with abnormal wave Ⅲ response threshold at 21 days and normal wave Ⅲ response threshold at 56 days,the latency of waves Ⅱ and Ⅲ,as well as the latency between Ⅰ-Ⅱ,Ⅱ-Ⅲ,and Ⅰ-Ⅲ were significantly higher,while the amplitude of wave Ⅲ and the ratio of Ⅲ/Ⅰ were significantly lower than those of in the control group and hypoxic group with normal wave Ⅲ response thresholds at 21 days and 56 days.The hearing loss of offspring mice in the hypoxia group was mainly bilateral.Conclusion Chronic intrauterine hypoxia can lead to varying degrees of hearing loss,but this type of hearing loss has a certain degree of retrogression in the early stages,and the body can undergo limited developmental compensation and repair.

ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ （AngⅡ） and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment， AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h， and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 （CCK-8） to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of B-type natriuretic peptide （BNP） in cell culture supernatant， and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently， the experiment was divided into a blank group （20% blank serum）， a model group （20% blank serum + 5×10^-5 mol·L^-1 AngⅡ）， low-， medium-， and high-dose （5%， 10% and 20%） serum containing Zhenwutang groups， an autophagy inhibitor group （1×10^-4 mol·L^-1 3-MA）， and autophagy inducer group （1×10^-7 mol·L^-1 rapamycin）. The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe （Lyso Tracker） and mitochondrial probe （Mito Tracker） co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect Caspase-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， and cytochrome C （Cyt C） in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase （Parkin）， phosphatase and tensin homolog （PTEN）-induced kinase 1 （PINK1）， and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3， cleaved Caspase-9， Bax， Bcl-2， and Cyt C in apoptosis-related pathways， phosphorylated ubiquitin ligase （p-Parkin）， phosphorylated PTEN-induced kinase 1 （p-PINK1）， p62， and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways， and the change of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10^-5 mol·L^-1， the cell activity was the lowest， and there was no cytotoxicity. At this concentration， the surface area of cardiomyocytes was significantly increased （P<0.01）， and the content of BNP in the supernatant of culture medium was significantly increased （P<0.05）. Therefore， AngⅡ with a concentration of 5×10^-5 mol·L^-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group， the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate （P<0.01） and significantly decreased mitochondrial membrane potential （P<0.01）. The results of immunofluorescence co-localization showed that compared with the blank group， the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group， the relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 in the model group was significantly up-regulated （P<0.01）， while the relative mRNA expression of Bcl-2， Parkin， and PINK1 was significantly down-regulated （P<0.01）. In addition， the relative protein expression of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 was significantly up-regulated （P<0.01）. The LC3Ⅱ/Ⅰ was significantly decreased， and the relative protein expression of Bcl-2， p-Parkin， p-PINK1， and Beclin1 was significantly down-regulated （P<0.01）. Compared with the model group， the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate （P<0.01）， and the decrease ratio of mitochondrial membrane potential is significantly lowered （P<0.01） in a dose-dependent manner. Additionally， both red and green fluorescence spots became more in these groups. In the 3-MA group， the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 was significantly down-regulated （P<0.05， P<0.01）， while that of Bcl-2， Parkin， and PINK1 was significantly up-regulated （P<0.01）. In the serum containing Zhenwutang groups， the relative protein expression levels of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 were significantly down-regulated （P<0.05，P<0.01）. The LC3Ⅱ/Ⅰ was significantly increased， and the relative protein expression levels of Bcl-2， p-Parkin， p-PINK1， and Beclin1 were significantly up-regulated （P<0.01）. ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.

Objective:To compare the dosimetric differences between a 3D-printed non-coplanar multi-channel applicator and traditional single-channel/co-planar multi-channel applicators in postoperative vaginal brachytherapy for early-stage endometrial cancer.Methods:CT scan data of 66 patients with stage I endometrial cancer, encompassing 100 3D brachytherapy CT imaging datasets, admitted to Department of Gynecologic Oncology of the Fourth Hospital of Hebei Medical University from December 2021 to June 2024 were retrospectively analyzed. Based on CT images and delineated structures, offline reconstructions of radiotherapy plans were performed for single-channel, coplanar multi-channel, and 3D-printed non-coplanar multi-channel applicators. These 3 radiotherapy plans were optimized, and the high-risk clinical target volume (HR-CTV) coverage (V 100 ≥90%) and doses to organs at risk (rectum, bladder) were compared. The prescription dose was standardized at 600 cGy, with constraints of rectal D 2 cm3 ≤420 cGy and bladder D 2 cm3 ≤480 cGy. Comparison among multiple groups was conducted by ANOVA. Bonferroni method was used to correct P-values for comparison between two groups. Results:When defined as HR-CTV D 90%≥600 cGy, bladder D 2 cm3 was (398.29±76.13)cGy and rectum D 2 cm3 was (402.10±49.77)cGy of the 3D-printed non-coplanar multi-channel group,which were significantly lower than those in the single-channel group [bladder D 2 cm3 (424.09±131.52) cGy, rectum D 2 cm3 (493.11±115.17) cGy] and coplanar group [bladder D 2 cm3 (461.28±134.84) cGy, rectum D 2 cm3 (508.75±119.02) cGy], respectively. When limiting bladder D 2 cm3≤480 cGy, rectal D 2 cm3 was (446.81±78.53 cGy) of the 3D-printed non-coplanar multi-channel group, which was significantly lower than those in the single-channel group [(589.71±153.91) cGy] and the coplanar group [(545.51±122.00) cGy], respectively. Meanwhile, HR-CTV V 100% (94.53%±3.42%) was higher than (91.19%±7.63%) in the coplanar group. When the rectal D 2 cm3 was ≤ 420 cGy, HR-CTV V 100% was (91.92%±4.04%) of the 3D-printed non-coplanar multi-channel group,which was significantly better than (79.23%±13.95%) in the single-channel group and (85.88%±6.86%) in the coplanar group, respectively. Conclusions:The 3D-printed non-coplanar multi-channel applicator significantly reduces bladder and rectal doses while enhancing target coverage, outperforming traditional single-channel and co-planar multi-channel applicators. This innovation provides an optimized solution for individualized precision radiotherapy.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

BACKGROUND:Apoptosis and autophagy imbalance in the hippocampal region of perimenopausal depressed rats are closely related to cognitive decline.Whether aerobic exercise can reduce apoptosis by promoting hippocampal autophagy and thus improve the learning and memory abilities of perimenopausal depressed rats is not clear.OBJECTIVE:To investigate the possible mechanism by which 4-week moderate-intensity treadmill exercise improves learning memory ability in ovariectomized stressed rats.METHODS:Forty Sprague-Dawely rats were randomly divided into four groups,namely,sham operation group(n=10),ovariectomized group(n=10),ovariectomized stress group(n=10)and ovariectomized stress exercise group(n=10).Except for the sham operation group,the ovaries were removed in the other three groups to establish a perimenopausal rat model,and then a depressed rat model was established by chronic unpredictable stress in the latter two groups.The rats in the ovariectomized stress exercise group underwent a 4-week moderate-intensity treadmill exercise.Tail suspension test and sucrose preference test were performed to text depression-like behaviors in rats after exercise and stress.The eight-arm maze experiment was used to test the learning and memory behaviors of rats after exercise and stress.Western blot was used to detect the protein levels of AMP-activated protein kinase/UNC-51 like autophagy activating kinase 1/mammalian target of rapamycin(AMPK/mTOR/ULK1),hippocampus apoptotic factor Caspase-3 and the protein expression of autophagy markers LC-3II/Beclin-1 in the hippocampus.RESULTS AND CONCLUSION:(1)Compared with the sham operation group,rats in the ovariectomized and ovariectomized stress groups had prolonged resting time in the tail suspension test and decreased sugar-water intake and sugar-water preference in the sucrose preference test.(2)Ovary removal reduced the learning memory capacity of rats,as evidenced behaviorally by a significant increase in the number of working memory errors,the number of reference memory errors,and the completion time,and an even more pronounced increase in the above measures in the ovariectomized stress group.(3)Compared with the ovariectomized group,there was a significant reduction in the number of working memory errors,the number of reference memory errors,and the completion time in the ovariectomized stress group.(4)Compared with the sham operation group,in the ovariectomized and ovariectomized stress groups,the expression of hippocampal apoptotic factor Caspase 3 protein was significantly elevated,the expression of autophagy-related factors proteins Beclin-1 and LC3II,as well as the protein expression of AMPK and ULK1,was decreased,whereas the expression of mTOR protein was elevated.Changes in the above indicators were more significant in the ovariectomized stress group.(5)Compared with the ovariectomized stress group,in the ovariectomized stress exercise group,the protein expression of Caspase 3 was significantly decreased,the protein expression of Beclin-1 and LC3II was significantly increased,the protein expression of AMPK and ULK1 was significantly increased,and the protein expression of mTOR was significantly reduced.To conclude,4-week moderate-intensity treadmill exercise may promote cellular autophagy and reduce apoptosis through the AMPK/mTOR/ULK1 autophagy signaling pathway,thereby enhancing the learning and memory capacity of rats with ovariectomized depression