OBJECTIVE: To identify and validate novel biomarkers for the early diagnosis of sepsis-associated acute kidney injury (SA-AKI) and precise continuous renal replacement therapy (CRRT) using proteomics. METHODS: A prospective multicenter cohort study was conducted. Patients with sepsis admitted to five hospitals in Taizhou City of Zhejiang Province from April 2019 to December 2021 were continuously enrolled, based on the occurrence of acute kidney injury (AKI). Sepsis patients were divided into SA-AKI group and non-SA-AKI group, and healthy individuals who underwent physical examinations during the same period were used as control (NC group). Peripheral blood samples from participants were collected for protein mass spectrometry analysis. Differentially expressed proteins were identified, and functional enrichment analysis was conducted on these proteins. The levels of target proteins were detected by enzyme linked immunosorbent assay (ELISA), and the predictive value of target protein for SA-AKI were evaluated by receiver operator characteristic curve (ROC curve). Additionally, sepsis patients and healthy individuals were selected from one hospital to externally verify the expression level of the target protein and its predictive value for SA-AKI, as well as the accuracy of CRRT treatment. RESULTS: A total of 37 patients with sepsis (including 19 with AKI and 18 without AKI) and 31 healthy individuals were enrolled for proteomic analysis. Seven proteins were identified with significantly differential expression between the SA-AKI group and non-SA-AKI group: namely cystatin C (CST3), β ₂-microglobulin (β ₂M), insulin-like growth factor-binding protein 4 (IGFBP4), complement factor I (CFI), complement factor D (CFD), CD59, and glycoprotein prostaglandin D2 synthase (PTGDS). Functional enrichment analysis revealed that these proteins were involved in immune response, complement activation, coagulation cascade, and neutrophil degranulation. ELISA results demonstrated specific expression of each target protein in the SA-AKI group. Additionally, 65 patients with sepsis (38 with AKI and 27 without AKI) and 20 healthy individuals were selected for external validation of the 7 target proteins. ELISA results showed that there were statistically significant differences in the expression levels of CST3, β ₂M, IGFBP4, CFD, and CD59 between the SA-AKI group and non-SA-AKI group. ROC curve analysis indicated that the area under the curve (AUC) values of CST3, β ₂M, IGFBP4, CFD, and CD59 for predicting SA-AKI were 0.788, 0.723, 0.723, 0.795, and 0.836, respectively, all exceeding 0.7. Further analysis of patients who underwent CRRT or not revealed that IGFBP4 had a good predictive value, with an AUC of 0.84. CONCLUSIONS Based on proteomic analysis, CST3, β ₂M, IGFBP4, CFD, and CD59 may serve as potential biomarkers for the diagnosis of SA-AKI, among which IGFBP4 might be a potential biomarker for predicting the need for CRRT in SA-AKI patients. However, further clinical validation is required.
Humans ; Sepsis/complications* ; Acute Kidney Injury/blood* ; Proteomics ; Prospective Studies ; Biomarkers/blood* ; Male ; Female ; beta 2-Microglobulin/blood* ; Middle Aged ; Cystatin C/blood* ; Aged

Objective: To investigate the distribution of HPA, HLA alleles and haplotypes among apheresis platelet donors in Changsha, China, and to establish an apheresis platelet donor database. Methods: High-resolution genotyping of HLA-A and -B was performed using PCR sequence based typing (SBT) and next generation sequencing (NGS). HPA genotyping was conducted using quantitative PCR (Q-PCR). The allele frequency, haplotype frequency and linkage disequilibrium parameters were calculated using the direct counting method, the maximum likelihood method and Arlequin software (V 3.1). Results: A total of 41 HLA-A alleles and 82 HLA-B alleles were detected, and 457 HLA-A-B haplotypes were found, of which 25 showed strong linkage disequilibrium (RLD>0.50). The HPA-3 and HPA-15 had the highest HPA polymorphism and antigen mismatch rate in apheresis platelet donor database in Changsha, and the dual antigen mismatch rate was 0.3704 and 0.3743, respectively. Conclusion: The polymorphism of apheresis platelet donor database in Changsha is complex and has strong regional characteristics. Establishing a high-resolution donor database will strongly support the provision of genetically matched platelets for clinical use, facilitating precise platelet transfusion therapy.

Glutamate-ammonia ligase (GLUL, also known as glutamine synthetase) is a crucial enzyme that catalyzes ammonium and glutamate into glutamine in the ATP-dependent condensation. Although GLUL plays a critical role in multiple cancers, the expression and function of GLUL in gastric cancer remain unclear. In the present study, we have found that the expression level of GLUL was significantly lower in gastric cancer tissues compared with adjacent normal tissues, and correlated with N stage and TNM stage, and low GLUL expression predicted poor survival for gastric cancer patients. Knockdown of GLUL promoted the growth, migration, invasion and metastasis of gastric cancer cells in vitro and in vivo, and vice versa, which was independent of its enzyme activity. Mechanistically, GLUL competed with β-Catenin to bind to N-Cadherin, increased the stability of N-Cadherin and decreased the stability of β-Catenin by alerting their ubiquitination. Furthermore, there were lower N-Cadherin and higher β-Catenin expression levels in gastric cancer tissues compared with adjacent normal tissues. GLUL protein expression was correlated with that of N-Cadherin, and could be the independent prognostic factor in gastric cancer. Our findings reveal that GLUL stabilizes N-Cadherin by antagonizing β-Catenin to inhibit the progress of gastric cancer.

Denosumab, a human monoclonal antibody, is used for the prevention of malignant tumor-related bone events and the treatment of osteoporosis with high fracture risk. Since its approval in China in 2019, denosumab-related osteonecrosis of the jaw (DRONJ) has attracted increasing attention. DRONJ, similar to bisphosphonate-related osteonecrosis of the jaw, often occurs after tooth extraction and manifests as exposed bone necrosis, purulent discharge, facial swelling and pain, severely impacting patients′ quality of life. However, the perioperative management strategies for DRONJ differ from those for bisphosphonate-related osteonecrosis of the jaw. This article summarizes the perioperative management strategies for high-risk DRONJ patients from aspects such as oral hygiene care, antibiotic use, drug discontinuation during the perioperative period, and surgical strategy selection, aiming to provide guidance for oral surgeons in managing tooth extraction in denosumab-treated patients.

Objective: To explore the impact of smart physical education assignment on physical health of male university students, so as to provide theoretical support and practical references for physical health improvement of male university students and implementing smart sports assignments. Methods: From September 2023 to January 2024, 317 sophomore male students from six Taekwondo elective classes at Hunan Institute of Engineering were selected and were randomly divided into an intervention group (n=157) and a control group (n=160). The intervention group was given sports assignments twice a week through smart means with an intervention duration of 15 weeks, each time for 25-35 minutes, in addition to the teaching according to the public course syllabus, while the control group was taught according to the public course syllabus. The physical and health indicators of both groups were tested before and after intervention,then the differences in various physical health indicators between two groups of students before and after intervention were compared through ttest and Mann-Whitney U test. Results: After the intervention, the vital capacity, 50 m run, sitandreach, 1 000 m run, and pullup scores of the intervention group significantly improved compared to those before intervention. The scores improved from (3 918.27±737.34)mL, 7.88(7.53,8.45)s, 9.80(2.70,15.75) cm, 4.30(4.12,4.50) min and 3.00(0.00,7.50) times to (4 574.19±800.61) mL, 7.65(7.37,8.12)s, 17.20(11.80,21.55)cm, 4.13(3.58,4.31)min and 5.00(1.00,10.00) times,respectively (t/Z=-7.60, 2.61, -8.39, 5.62, -2.72, P<0.05). Before intervention, there was no statistically significant difference in physical health indicators between the intervention group and the control group (P>0.05).After intervention,the scores of the intervention group on the vital capacity,50 m run,sitandreach,1 000 m run and pullup, were significantly higher than those of the control group [(4 310.97±808.90)mL, 7.75(7.40,8.30)s, 14.10(8.42,17.87)cm, 4.29(4.08,4.45)min and 4.00(1.00,7.00) times] (t/Z=2.91, -4.55, -4.75, -4.15, 2.58, P<0.05). Conclusions Having 25-35 min smart physical education assignment twice a week can effectively improve physical health level of male college students. It is recommended to assign appropriate amount of smart sports homework to improve physical health level of college students, while ensuring the amount and intensity of physical activity in public physical education courses.

  Objective  To explore the effect of esketamine on working memory impairment in neuropathic mice and its underlying mechanism.  Methods  Fifty clean grade male C57BL/6J mice (2 months) were divided into 5 groups by random number table method: sham+saline (SN group), CCI+saline (CN group), CCI+esketamine(CE group), CCI+ANA-12 (CA group), CCI+ANA-12+esketamine (CAE group), with 10 mice in each group. Chronic constriction injury (CCI) was employed to establish a neuropathic pain model. On the 16th day after modeling, CE group and CAE group were administered ketamine (10 mg/kg), CAE group received ANA-12 (0.5 mg/kg) half an hour before ketamine injection, CA group was only given ANA-12, and SN group and CN group received an equivalent volume of saline. The administration was done through intraperitoneal injection for 5 consecutive days. The open-field test (OFT), paw withdrawal threshold (PWT), paw withdrawal latency (PWL) and Y-maze test were performed from day 21 after surgery. Bromo-2-deoxyUridine (BrdU) was dissolved in saline and intraperitoneally injected into the mice on days 21 to 23 after the surgery. Western blot was performed to determine the expression of Brain-derived neurotrophic factor(BDNF) in hippocampus, the immunofluorescence was performed to determine the number of BrdU and doublecortxin (DCX) positive cells in the dentate gyrus (DG) area of hippocampal.  Results  Compared with SN group, the other four groups showed significant reductions in both PWT and PWL on day 21 after surgery (all P<0.05); There was no significant difference in the total distance travelled by the 5 groups of mice (P=0.142); In the Y maze test, compared to SN group, the accurate percentage of spontaneous alternation in CN group showed significant reductions (P<0.001), which was reversed by esketamine administration (P<0.001); compared with CE group, there was a significant reduction of the accurate percentage of spontaneous alternation in CAE group (P=0.004). The expression of BDNF protein in CN group was lower than that in SN group (P=0.021) and CE group (P=0.030), and compared with CE group, the expression of BDNF was significant decreased in CEA group (P=0.043). The number of BrdU positive and DCX positive cells significantly reduced in the DG area of the hippocampus in CN group compared to SN group (P=0.025) and CE group (P=0.003). The number of BrdU positive and DCX positive cells in CAE group significantly reduced in the DG region of the hippocampus compared to CE group (P=0.014).  Conclusion  Esketamine maybe improve working memory impairment in neuropathic mice and the neurogenesis in dentate gyrus area of hippocampal through the BDNF-TrkB pathway.

Objective:To evaluate the relationship between the mechanism underlying the antidepressant effect of S-ketamine and hippocampal gamma-aminobutyric acid B receptor (GABA BR) in mice. Methods:A total of 54 male C57BL/6(B6) mice, aged 8 weeks, weighing 25-30 g, were used in this study. Forty mice were selected to develop the depression model by chronic social defeat stress. Twenty-six depression-susceptible mice were screened out by social avoidance test at day 11 after developing the model and divided into 2 groups ( n=13 each) by a random number table method: depression-susceptible group (Sus group) and depression-susceptible + S-ketamine group (Sus + S-ket group). The remaining 14 mice served as control group (C group). Starting from day 12 after developing the model, S-ketamine 10 mg/kg was intraperitoneally injected every day for 3 consecutive days in Sus+ S-ket group, while the equal volume of normal saline was given instead in C group and Sus group. The open field test was performed at 1 h after the last administration, and the total distance of movement was recorded. The forced swimming test was performed at 1 day after the open field test, and the immobile time was recorded. The sucrose preference test was performed to calculate the proportion of sucrose consumption at 1 day after the forced swimming test. One hour after the end of behavioral test, mice were sacrificed, and the hippocampal tissues were removed. Western blot was used to detect the expression of GABA BR1, GABA BR2, mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), brain-derived neurotrophic factor (BDNF), tyrosine kinase receptor B (TrkB), phosphorylated TrkB (p-TrkB), glutamate receptor 1 (GluR1) and postsynaptic dense protein 95 (PSD95). The p-mTOR/mTOR ratio and p-TrkB/TrkB ratio were calculated. The fluorescence intensity of BDNF in hippocampal CA1 region was detected by immunofluorescence. The number of dendritic spines in hippocampal CA1 region was measured by Golgi staining. Results:In the open field test, no statistically significant difference in the total distance was detected among the three groups ( P>0.05). Compared with C group, the immobile time in the forced swimming test was significantly prolonged, the proportion of sucrose consumption was decreased, the expression of hippocampal GABA BR1, GABA BR2, BDNF, GluR1 and PSD95 was down-regulated, and the ratios of p-mTOR/mTOR and p-TrkB/TrkB were decreased, the fluorescence intensity of BDNF and total number of dendritic spines in the hippocampal CA1 region were decreased in Sus group ( P<0.05), and no significant change was found in the parameters mentioned above in Sus+ S-ket group ( P>0.05). Compared with Sus group, the immobile time in the forced swimming test was significantly shortened, the proportion of sucrose consumption was increased, the expression of hippocampal GABA BR1, GABA BR2, BDNF, GluR1 and PSD95 was up-regulated, the ratios of p-mTOR/mTOR and p-TrkB/TrkB were increased, and the fluorescence intensity of BDNF and total number of dendritic spines in the hippocampal CA1 region were increased in Sus+ S-ket group ( P<0.05). Conclusions:The mechanism underlying the antidepressant effect of S-ketamine may be related to up-regulation of hippocampal GABA BR expression, activation of mTOR-BDNF signaling pathway, and improvement in synaptic plasticity in mice.

ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ²=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ²=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ²=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4⁺， and CD4⁺/CD8⁺ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19^-， HLA/DR⁺， and CD25⁺ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4⁺， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.

Objective:To investigate the effect and mechanism of non-selective histone deacetylase (HDAC) inhibitor sodium butyrate (NaB) on neuropathic pain and pain-induced memory impairment in mice.Methods:Forty clean grade male C57BL/6J mice were were divided into 4 groups by random number table method ( n=10 in each group): sham + saline, sham + NaB, chronic constriction injury (CCI)+ saline and CCI + NaB.The mouse CCI model was established by sciatic nerve ligation. Non-selective HDAC inhibitors NaB(300 mg/kg) was intraperitoneally injected into the mice in Sham+ NaB group and CCI+ NaB group once a day 15-28 days after modeling, while the mice in Sham+ saline group and CCI+ saline group were intraperitoneally injected with the same volume of saline. On the 14th and 28th day after operation, the athletic ability was measured by open field test (OFT), the pain behavior was measured by paw withdrawal threshold (PWT) and paw withdrawal latency (PWL), and the memory function was measured by Y-maze. After the behavioral experiment, hippocampus and spinal dorsal horn tissues were taken for the activity of HDAC measurement, and hippocampus tissues were taken for the expression levels of BDNF and PSD95 measurement. SPSS 25.0 software was used for statistical analysis. The data were compared by repeated measurement ANOVA and one-way ANOVA. Results:After treatment with NaB, the interaction effects of the accuracy of spontaneous alternation of PWT, PWL and Y maze in mice were significant( F=21.07, 6.98, 7.79, all P<0.05). Compared with the Sham + saline group, the PWT((0.83±0.30)g, (0.25±0.22)g, (0.24±0.11)g; both P<0.05), the PWL((14.97±4.02)s, (5.99±1.51)s, (6.87±0.90)s; both P<0.05) and the spontaneous alternation in Y maze(71.57±2.80)%, (56.96±0.60)%, (62.86±4.94)%; both P<0.05) in CCI+ Saline group and CCI+ NaB group were lower. After treatment with NaB, compared with CCI + saline group, PWT((0.22±0.13)g, (0.62±0.23)g; P<0.05), PWL((5.62±2.00)s, (8.82±2.13)s; P<0.05)and the accuracy of spontaneous alternation of Y maze were significantly higher ((56.54±7.50)%, (66.35±8.20)%; P<0.05), the HDAC activity in hippocampus((173.40±7.38)%, (122.70±8.40)%; P<0.05)and in spinal cord ((153.40±10.58)%, (111.40±11.40)%; P<0.05)were significantly lower, and the expression of BDNF((0.65±0.06), (0.87±0.43); P<0.05)and PSD95((0.70±0.40), (0.87±0.04); P<0.05)were significantly higher in CCI + NaB group. Conclusion:NaB can improve neuropathic pain and pain-induced memory impairment.The mechanism may be related to the inhibition of HDAC activity and the up-regulation of BDNF and PSD95 expression in hippocampus.

Objective To explore the application of urodynamics in the diagnosis and treatment of nonmonosymp-tomatic nocturnal enuresis(NMNE)in children,and to provide some evidence for its clinical treatment. Methods Patients who were clinically diagnosed as NMNE at Department of Pediatrics Kidney Rheumatism and Pediatric Urology of Shengjing Hospital of China Medical University from January 2015 to August 2016 were collected. Sixty - six children with NMNE were included in the urodynamic study,and the different treatments were selected according to the results of the urodynamics and the efficacy was evaluated. Results The urodynamic results of 61 patients(92. 43％)were abnor-mal,and 5 cases(7. 57％)had no abnormalities. There were 35 cases with overactive bladder(OAB)in abnormal re-sults,with a remarkable decrease in bladder pressure capacity in 7 cases,detrusorsphincter discoordination(DSD)in 1 case,and OAB with other abnormal results in 18 cases. OAB was found in 53 cases,and the detection rate was 86. 89％(53 / 61 cases),accounting for 80. 3％(53 / 66 cases)of the total samples. The desmopressin acetate combined with Tolterodine tartrate was used to treat OAB patients,and its total effective rate was 94. 34％(50 / 53 cases). The biofeed-back treatment had effect on DSD child. The patients with a marked decrease in bladder pressure capacity and with the normal urodynamics received desmopressin acetate joint behavior training treatment,and the efficiency of the therapy was 71. 43％(5 / 7 cases)and 80. 00％(4 / 5 cases),respectively. Conclusions The urodynamics study can uncover the pathophysiological mechanism of NMNE,so this examination is essential for NMNE children to improve the treat-ment. Since the rate of OAB in NMNE is very high,it is suggested that anticholinergic medications combined with desmopressin can be used in the initial treatment of the NMNE patients to improve the rate of cure and prognosis.