BACKGROUND:Erythropoietin has anti-inflammatory,anti-apoptotic,and pro-bone defect repair effects.To date,fewer studies have been conducted on its effects and molecular mechanism underlying restorative dentin formation after pulp injury. OBJECTIVE:To explore the effect of erythropoietin on restorative dentin formation after pulp injury. METHODS:(1)Animal experiment:Thirty-two rats were randomly divided into control group(n=16)and experimental group(n=16).In the experimental group,collagen sponges containing erythropoietin were used to directly cap the pulp at the pulp injury,and in the control group,collagen sponges containing PBS were used to directly cap the pulp at the exposed pulp injury.The cavity was then closed with glass ionomer adhesive.After 2 and 4 weeks of treatment,the maxillary bones of the two groups were collected,and the expression of nestin in dentin was detected by immunohistochemistry,and the reparative dentin production was observed by hematoxylin-eosin staining.The maxillae of four Sprague-Dawley rats were taken for immunohistochemical detection of erythropoietin expression in molar and incisor teeth.(2)Cell experiment:Human dental pulp cells,human periodontal ligament cells and human gingival fibroblasts were obtained from human dental tissue,periodontal ligament,and gingival tissue.Real-time reverse transcription PCR(RT-PCR)was used to detect the mRNA expression of erythropoietin.Erythropoietin,dentin sialophosphoprotein,dentin matrix protein 1,and nestin mRNA levels in human pulp cells were detected by RT-PCR under induced or uninduced odontoblastic differentiation.After down-regulation of erythropoietin expression or exogenous administration of erythropoietin intervention under induced or uninduced differentiation odontoblastic differentiation,the relative mRNA expression of dentin sialophosphoprotein and dentin matrix protein 1 in human pulp cells was detected by RT-PCR,and the formation of mineralized nodules was detected by alizarin red S staining,and mRNA and protein expressions of bone morphogenetic protein 2 were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:(1)Animal experiment:Compared with the control group,the restorative dentin production and nestin expression were higher in the experimental group after 2 and 4 weeks of treatment.The expression of erythropoietin was weakly positive in pulp,odontoblastic cell layer and periodontal membrane of the rat's first maxillary molar,and strongly positive in odontoblasts.(2)Cell experiment:The mRNA expression of erythropoietin was higher in human dental pulp cells than in the other two types of cells.The mRNA expressions of dentin sialophosphorin,dentin matrix protein 1,nestin,erythropoietin and bone morphogenetic protein 2 in human pulp cells increased and the formation of mineralized nodules during odontoblastic differentiation under induction compared with non-induction conditions.The mRNA expression of dentin sialophosphoprotein,dentin matrix protein 1,nestin,bone morphogenetic protein 2 and the formation of mineralized nodules were decreased in human pulp cells after downregulation of erythropoietin under induced odontoblastic differentiation,and the protein expression of bone morphogenetic protein 2 was also decreased.After exogenous erythropoietin intervention,the expression of the above indexes in human dental pulp cells increased.To conclude,erythropoietin can promote the formation of dentin to some extent.

OBJECTIVES: To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease (DKD) induced by advanced glycation endproducts (AGEs). METHODS: Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline, Guijianyu extract at 3 doses or irbesartan for 12 weeks, and the changes in renal pathology and structure were observed using transmission electron microscopy, and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry. In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h, the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay; Western blotting was performed to detect the protein expressions of RAGE, VEGFA, TNF-α, NF-κB(p65), IL-6 and caspase-3, and the mRNA expressions of RAGE, NF-κB (p65), VEGFA and IL-6 were detected with RT-qPCR. RESULTS: In mouse models of DKD, treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE, VEGFA, NF-κB(p65), and IL-6 proteins and the mRNA expressions of RAGE, NF-κB, and IL-6. In MPC-5 cells, exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE, NF‑κB (p65), VEGFA, TNF-α, IL-6 and caspase-3 (P<0.05) and mRNA expressions of RAGE, NF-κB (p65), VEGFA, and IL-6. Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE, NF-κB(p65), VEGFA, TNF-α, IL-6, and caspase-3. Furthermore, Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE, VEGFA, TNF-α, IL-6, and caspase-3 and mRNA expressions of RAGE, NF-κB (p65), IL-6, and VEGFA in MPC-5 cells. CONCLUSIONS Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.
Animals ; Glycation End Products, Advanced ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Signal Transduction/drug effects* ; Podocytes/pathology* ; Diabetic Nephropathies/drug therapy* ; Receptor for Advanced Glycation End Products ; Vascular Endothelial Growth Factor A/metabolism* ; Interleukin-6/metabolism* ; Male

Currently, transcatheter intervention has emerged as a first-line treatment for coarctation of the aortic. Due to the radiation exposure associated with catheter interventional therapy, there are numerous restrictions, which harms both patients and medical personnel and is dependent on sizable radiation apparatus. Here, we report for the first time a case of echo-guiding percutaneous aortic stent implantation for a 27 years female patient of reproductive age. After discharge, the patient's aortic coarctation pressure decreased to 18 mm Hg, and the surgical results were satisfactory.

Objective:To explore the characteristics of phenylalanine hydroxylase ( PAH) gene variants and prenatal diagnosis for 43 Chinese pedigrees affected with Phenylketonuria (PKU). Methods:Forty three PKU pedigrees diagnosed at the First Affiliated Hospital of Zhengzhou University between 2019 and 2021 were selected as the study subjects. Variants of the PAH gene of the probands were screened by high-throughput sequencing, and candidate variants were verified by Sanger sequencing. Negative cases were further analyzed by multiplex ligation-dependent probe amplification (MLPA) to detect large fragment deletions and duplications of the PAH gene. For 43 women undergoing subsequent pregnancy, Sanger sequencing, MLPA, combined with short tandem repeats (STR) sequence-based linkage analysis, were carried out for prenatal diagnosis. Results:Among the 86 alleles carried by the 43 probands, 78 nucleotide variants (90.70%) and 3 large deletions (3.49%) were found based on high-throughput sequencing and MLPA. The 81 mutant alleles had included 21 missense variants, 5 splice site variants, 4 nonsense variants, 2 microdeletions, 1 insertional variant and 2 large fragment deletions. Relatively common variants have included p. Arg243Gln (23.26%), p. Arg111Ter (8.14%), EX6-96A>G (6.98%), p. Val399Val (5.81%) and p. Arg413Pro (4.65%). Most of the variants were located in exons 7, 11, 3, 6 and 12. For the 43 families undergoing prenatal diagnosis, 9 fetuses (20.45%) were diagnosed with PKU, 20 (45.45%) were heterozygous carriers, and 15 (34.09%) did not carry the same pathogenic allele as the proband. All neonates were followed up till 6 months old, and the accuracy of prenatal diagnosis was 100%.Conclusion:The combination of high-throughput sequencing, Sanger sequencing, MLPA and linkage analysis can increase the diagnostic rate of PKU and attain accurate prenatal diagnosis.

Objective·In view of the low funding rate of the Young Scientists Fund of National Natural Science Foundation of China in the medical field,this research conducted an in-depth investigation of the applicants in a medical school and quantitatively analyzed the influencing factors of project establishment,so as to provide reference for further improving the quality of project application and strengthening the training of young medical talents.Methods·A cross-sectional survey was used to select applicants from secondary schools of Shanghai Jiao Tong University School of Medicine and its 13 affiliated hospitals who had experience in applying for the fund from 2020 to 2022.Logistic stepwise regression model was applied to modeling and analysis by using the project application result as the dependent variable and the applicants'basic information and application status as independent variables.Results·The analysis results of 921 applicants showed that educational background(OR=1.86,95%CI 1.14?3.04),graduate university(OR=2.45,95%CI 1.47?4.08),sufficient preliminary work foundation(OR=4.22,95%CI 2.44?7.29),average impact factor of representative works(OR=1.10,95%CI 1.04?1.17),and total self-evaluation score of application documents(OR=1.06,95%CI 1.04?1.08)were the main influencing factors for project approval.The average impact factor and the highest impact factor of the approved representative works showed an increasing trend year by year.In addition,the influencing factors for the approval of young doctors and full-time researchers were different.The longer the graduation time of young doctors,the lower the approval rate.Conclusion·The funding rate of Young Scientists Fund is low,and the requirements for scientific research achievements are increasing year by year.Attention should be paid to early accumulation and improving the quality of application writing.Further measures should be taken to strengthen the early training of young medical talents,improve cultivation measures and consolidate research foundations.

Objective To investigate the effect of fibulin-3 on the senescence of intervertebral disc nucleus pulposus cells(NPCs)through the regulation of tissue inhibitor of metalloproteinases 3(TIMP-3)expression and to elucidate the molecular mechanisms involved.Methods 1).The nucleus pulposus tissues and imaging data of 37 patients who had undergone intervertebral disc surgery were collected.The degree of degeneration of the intervertebral discs were classified according to the Pfirrmann grading system.The senescence degree of NPCs was determined using senescence-associated β-galactosidase(SA-β-gal)staining.Fibulin-3 expression levels were determined using Western blot and ELISA.The relationship between fibulin-3 and disc degeneration and NPCs senescence was investigated.2).Human intervertebral disc NPCs were cultured in vitro.The proliferation and senescence of NPC across continuous passage were observed via CCK-8 assay and SA-β-gal staining,respectively.Fibulin-3 expression levels and the expression of inflammatory cytokines and matrix metalloproteinases were assessed.Exogenous fibulin-3 was added to verify its effect on the proliferation and senescence of NPCs.3).The effect of fibulin-3 on the apoptosis and proliferation of NPCs was verified through gene overexpression,which was used in combination with an apoptosis inhibitor for bidirectional verification.4).Bioinformatics analysis was performed to explore the relationship between fibulin-3 and the TIMP family.Experiments overexpressing fibulin-3 and silencing the TIMP-3 gene were performed to verify their role in NPCs senescence.Results 1).The intervertebral disc degeneration samples from 37 patients were classified according to the Pfirrmann grading system.The higher the degeneration grade,the lower fibulin-3 expression.Spearman correlation analysis showed that the disc grade was negatively correlated with the NPC senescence grade(r=-0.87,P＜0.001)and fibulin-3 expression(r=-0.79,P＜0.001).2).As the passage number of NPCs increased,fibulin-3 expression gradually decreased,cell proliferation ability weakened,and the expression of inflammatory cytokines and matrix metalloproteinases increased.After exogenous fibulin-3 was added,cell morphology and growth status were maintained,cell senescence was significantly inhibited,and the expression of inflammatory cytokines and matrix metalloproteinases was markedly reduced.3).Gene overexpression experiments showed that fibulin-3 reduced NPC apoptosis and promoted cell proliferation,thereby inhibiting NPC senescence.4).Bioinformatics analysis revealed a significant association between fibulin-3 and TIMP-3 of the TIMP family.Further experiments confirmed that overexpressing fibulin-3 enhanced TIMP-3 expression,while silencing the TIMP-3 gene significantly weakened the inhibitory effect of fibulin-3 on NPCs senescence.This indicates that,through regulating TIMP-3,fibulin-3 inhibits the activity of matrix metalloproteinases,affects the synthesis and degradation of the extracellular matrix,and ultimately inhibits NPCs senescence.Conclusion This study demonstrates that fibulin-3 plays a crucial role in inhibiting the senescence of intervertebral disc NPCs by regulating TIMP-3.The specific mechanisms involved are as follows,fibulin-3 upregulates TIMP-3 expression,inhibits matrix metalloproteinase activity,and reduces extracellular matrix degradation,thereby promoting extracellular matrix synthesis.Additionally,fibulin-3 inhibits NPCs senescence by reducing apoptosis and promoting cell proliferation.Therefore,fibulin-3 and TIMP-3 have potential therapeutic significance in maintaining intervertebral disc health and delaying degeneration.

Antibody-drug conjugate (ADC) has the characteristics of low toxicity and high efficiency, and plays an important role in cancer treatment. However, due to the complexity of its structure, it brings difficulties in pharmacokinetic (PK) bioanalysis. This study established an analytical method for the detection of ADC (RC108) in cynomolgus monkey plasma by ligand-binding assay (LBA) and liquid chromatography tandem mass spectrometry (LC-MS/MS), which was used to analyze and quantify the total antibody, bound antibody and free drug in cynomolgus monkey plasma. Based on the LBA method, rabbit anti-RC108 Fab and mouse anti-MMAE (monomethyl auristatin E) mAb were pre-coated in 96-well plates as the total antibody and antibody binding reagents, respectively. The samples to be tested were added, and then the detection reagents were added in turn. Goat anti-human IgG (H+L)-HRP, chromogenic solution tetramethylbenzidine (TMB), H₂SO₄ terminate the reaction, read data at 450 nm/630 nm wavelength of microplate reader; LC-MS/MS analysis method quantifies MMAE concentration, and refer to relevant regulations for methodological validation. The analytical method for quantifying total antibody, bound antibody and free drug of RC108 drug obtained good accuracy and precision, and the selectivity, dilution linearity, hook effect, parallelism and stability were verified. Meet the requirements of biological analysis. Finally, a bioanalytical method for the determination of the concentration of the test substance RC108 (total antibody, conjugated antibody, free MMAE) in cynomolgus monkey plasma with high sensitivity and high throughput was established by LBA and LC-MS/MS method. Subsequent non-clinical research on PK research in cynomolgus monkeys will provide technical support.

In recent years, monotherapy and combination therapy with immune checkpoint inhibitors (ICIs) have achieved good efficacy in a variety of malignancies from solid tumors to lymphomas and have become a standardized and systematic treatment modality for many cancers. However, there is still a lack of studies on the safety of ICIs in hepatitis B virus (HBV)-infected patients with malignancies, and early studies have reported HBV reactivation due to ICI antitumor therapy in clinical practice. With reference to related literature, this article reviews the recent clinical trials and application of ICIs in cancer patients with chronic viral infection and clarifies the efficacy and safety of ICIs in this special population, in order to provide a reference for clinical medication.

Objective    To explore the short-term efficacy and safety of pembrolizumab combined with chemotherapy in the neoadjuvant treatment of non-small cell lung cancer. Methods    The clinical data of 11 male patients with non-small cell lung cancer who underwent pembrolizumab combined with neoadjuvant chemotherapy in the Department of Thoracic Surgery, the First Affiliated Hospital of Xi'an Jiaotong University from December 2019 to June 2021 were retrospectively analyzed. The average age of the patients was 52.0-79.0 (62.0±6.9) years. The imaging data and pathological changes before and after neoadjuvant treatment were compared, and adverse reactions during neoadjuvant treatment were recorded. Objective remission rate (ORR) and main pathological remission rate (MPR) and pathological complete remission rate (pCR) were the main observation endpoints. Results    After preoperative neoadjuvant therapy with pembrolizumab combined with platinum or paclitaxel, all patients successfully underwent thoracoscopic radical resection of lung cancer. The ORR was 72.7%, and the MPR was 81.8%. Among them, 45.5% of patients achieved pCR. The main adverse reactions were hypoalbuminemia, decreased appetite and nausea. The mortality rate within 30 days after surgery was 0, and no tumor metastasis was observed. Conclusion    Pembrolizumab combined with neoadjuvant chemotherapy is safe and feasible to treat non-small cell lung cancer, and the short-term efficacy is beneficial.

Objective     To investigate the safety and feasibility of day surgery for patients with palmar hyperhidrosis based on the principles of enhanced recovery after surgery (ERAS). Methods     We retrospectively reviewed the medical records of consecutive patients who underwent endoscopic thoracic sympathicotomy (ETS) in the First Affiliated Hospital of Xi'an Jiaotong University from March 2020 to December 2021. Patients were divided into a day surgery group and a conventional group according to their perioperative management methods. The patients in the day surgery group underwent an optimized perioperative procedure under the guidance of ERAS, and were ventilated with a laryngeal or face mask during the operation. The patients in the conventional group completed the preoperative examination, operation and postoperative observation according to the conventional procedures, and were intubated with a single-lumen endotracheal tube. The demographic characteristics, operation time, hospital stay, postoperative complications, and hospitalization cost were compared between the two groups. Results     Finally 172 patients were collected, including 90 males and 82 females, with an average age of 25.97±7.43 years. There were 86 patients in each group. All patients ceased suffering from palmar sweating after surgery. No patient experienced massive bleeding or conversion to thoracotomy. There was no statistical difference in operation time between the two groups (P=0.534). Patients in the day surgery group were discharged within 24 hours. The average hospital stay in the conventional group was 2.09±0.41 days. Incidence of postoperative respiratory complications, and the hospitalization cost of the day surgery group were significantly lower than those of the conventional group (P<0.001). The satisfaction rate in both groups was greater than 95%. Conclusion     Day surgery for patients with palmar hyperhidrosis based on the principles of ERAS is safe and feasible, which can reduce postoperative complications, shorten the length of hospital stay and save the cost of hospitalization.