[摘 要] 目的：探讨环状RNA hsa_circ_0046701在胶质瘤组织中的表达及其对胶质瘤U251细胞增殖、迁移与侵袭的影响。方法：收集2022年6月至2023年3月期间在同济大学附属普陀人民医院接受手术治疗的52例胶质瘤患者的瘤组织标本及临床资料，另收集30例正常脑组织标本作为对照。通过qPCR法检测胶质瘤组织中hsa_circ_0046701表达水平，分析其与患者临床特征间的关系，通过Kaplan-Meier法分析hsa_circ_0046701水平与生存预后的关系。利用RNA干扰技术，分别将circ_0046701过表达及空载体（vector）、siRNA-circ_0046701及阴性对照（si-NC）质粒转染到胶质瘤U251细胞中，实验分为si-circ_0046701组、si-NC组、circ_0046701 OE组、Vector组。应用CCK-8法、Transwell小室实验检测各组细胞的增殖、迁移及侵袭能力，WB法检测各组细胞中vimentin、Snail、E-cadherin和cyclin D1蛋白的表达。结果：胶质瘤组织中hsa_circ_0046701表达显著高于正常脑组织（P < 0.01）。hsa_circ_0046701高表达组患者WHO脑胶质瘤分级（Ⅲ~Ⅳ级）占比显著高于低表达组（P < 0.01），其高表达组患者术后生存期显著短于低表达组。与si-NC组相比，si-circ_0046701组U251细胞的增殖能力显著降低（P < 0.05或P < 0.01）、迁移及侵袭细胞数均显著减少（均P < 0.01），细胞中vimentin、Snail、cyclin D1蛋白表达均明显降低（均P < 0.01）、E-cadherin蛋白表达明显增高（P < 0.01）；与Vector组相比，circ_0046701 OE组U251细胞的增殖能力显著升高（P < 0.01）、迁移及侵袭细胞数均显著增多（均P < 0.01），细胞中vimentin、Snail、cyclin D1蛋白表达均显著增高（均P < 0.01）、E-cadherin蛋白表达明显降低（P < 0.01）。结论：环状RNA hsa_circ_0046701在胶质瘤组织中呈高表达，并与患者的不良预后密切相关；敲低hsa_circ_0046701表达能够抑制脑胶质瘤U251细胞的增殖、迁移及侵袭能力。

Objective: To explore the clinical features, treatment and prognosis of emphysematous pyelonephritis (EPN), so as to enhance the clinical awareness of this disease. Methods: A retrospective analysis was conducted on the clinical data of one EPN patient at The Second Affiliated Hospital, Guangzhou University of Chinese Medicine, and a literature review was performed on articles published in the China National Knowledge Infrastructure and PubMed databases from Jan.1, 2015 to Dec.31, 2024. Results: The patient, a 62-year-old male with a 5 years' history of type 2 diabetes, was admitted due to left flank pain for 4 days, with a temperature of 39.4 ℃.Laboratory tests indicated significantly elevated inflammatory markers, decreased platelet count, and abnormal coagulation function.Preoperative blood and urine cultures showed positivity for Escherichia coli.Computed tomography (CT) revealed complete erosion of the left kidney, with gas in the left ureter and surrounding effusion, as well as multiple free gas in the abdominal cavity, bilateral ureteral stones, right renal lower calyx stones.After a multidisciplinary consultation, he underwent emergency phase Ⅰ left pyeloplasty and perirenal drainage with ureteral stenting.After discharge, the patient received maintenance hemodialysis once every two days in the outpatient clinic.One week after-discharge, the patient was readmitted due to polypnea.Following symptomatic management, vital signs stabilized.Approximately 2 months after the first-stage surgery, ureteroscopic stone extraction was successfully performed.One month after the stone extraction procedure, a follow-up CT showed normalization of the left kidney, renal pelvis and calyces, leading to phase Ⅱ laparoscopic left nephrectomy via the abdominal approach, with postoperative pathology indicating renal necrosis.Among 89 EPN patients reported in 35 articles, the median age was 58(24-92) years old；there were 59（66.3%） females and 30（33.7%） males；fever was the most common clinical symptom (60.7%)；73(82.0%) had diabetes, 12 (13.5%) had urinary tract obstruction；55 (61.8%) were infected with Escherichia coli, and 7 (7.9%) were infected with Klebsiella pneumoniae; 13 died due to ineffective treatmen. Conclusion: EPN presents acutely and progresses rapidly, often leading to misdiagnosis due to the lack of specific early symptoms.Abdominal CT is the preferred imaging modality for rapid diagnosis, and proactive interdisciplinary intervention can improve survival rates, reduce the need for nephrectomy, and enhance prognosis.

Dental stem cells (DSCs), a distinct subset of mesenchymal stem cells (MSCs), are isolated from dental tissues, such as dental pulp, exfoliated deciduous teeth, periodontal ligament, and apical papilla. They have emerged as a promising source of stem cell therapy for tissue regeneration and autoimmune disorders. The main types of DSCs include dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHED), periodontal ligament stem cells (PDLSCs), and stem cells from apical papilla (SCAP). Each type exhibits distinct advantages: easy access via minimally invasive procedures, multi-lineage differentiation potential, and excellent ethical acceptability. DSCs have demonstrated outstanding clinical efficacy in oral and maxillofacial regeneration, and their long-term safety has been verified. In oral tissue regeneration, DSCs are highly effective in oral tissue regeneration for critical applications such as the restoration of dental pulp vitality and periodontal tissue repair. A defining advantage of DSCs lies in their ability to integrate with host tissues and promote physiological regeneration, which render them a better option for oral tissue regenerative therapies. Beyond oral applications, DSCs also exhibit promising potential in the treatment of systemic diseases, including type Ⅱ diabetes and autoimmune diseases due to their immunomodulatory effects. Moreover, extracellular vesicles (EVs) derived from DSCs act as critical mediators for DSCs' paracrine functions. Possessing regulatory properties similar to their parental cells, EVs are extensively utilized in research targeting tissue repair, immunomodulation, and regenerative therapy-offering a "cell-free" strategy to mitigate the limitations associated with cell-based therapies. Despite these advancements, standardizing large-scale manufacturing, maintaining strict quality control, and clarifying the molecular mechanisms underlying the interaction of DSCs and their EVs with recipient tissues remain major obstacles to the clinical translation of these treatments into broad clinical use. Addressing these barriers will be critical to enhancing their clinical applicability and therapeutic efficacy. In conclusion, DSCs and their EVs represent a transformative approach in regenerative medicine, and increasing clinical evidence supports their application in oral and systemic diseases. Continuous innovation remains essential to unlocking the widespread clinical potential of DSCs.
Humans ; Dental Pulp/cytology* ; Translational Research, Biomedical ; Mesenchymal Stem Cells/cytology* ; Periodontal Ligament/cytology* ; Stem Cells/cytology* ; Regeneration ; Tooth, Deciduous/cytology* ; Cell Differentiation ; Tissue Engineering/methods* ; Regenerative Medicine

Objective: To establish and validate a whole blood (WB) supply model, thereby providing practical experience for the clinical application of WB in domestic trauma emergency care and informing the development of a wartime blood supply system for the military. Methods: A “10×24” WB supply model was established by formulating blood collection protocols, storage standards, and transfusion criteria. Multiple WB samples were tested under specific storage conditions to assess key indicators at different time points, including red blood cell (RBC), white blood cell (WBC), and platelet counts, hemoglobin concentration, coagulation parameters (PT, APTT, TT, FIB), coagulation factor activity, thromboelastography (TEG) parameters, and electrolyte levels. Additionally, clinical data from hemorrhagic patients who met the criteria for WB transfusion and were admitted between March and July 2024 were analyzed to evaluate WB transfusion volume. Results: RBC counts and hemoglobin levels remained stable in WB stored at 4℃ for up to 10 days. However, platelet counts and coagulation function (PT, APTT) significantly declined with prolonged storage, while potassium levels increased. From March to July 2024, the model was successfully applied to 23 patients with acute hemorrhage, with a median WB transfusion volume of 543 mL. A detailed case study of a severe traumatic hemorrhagic shock patient was reported, who was successfully treated with 5.5 units of refrigerated WB combined with component blood. Conclusion: The “10×24” WB supply model demonstrated acceptable changes in critical quality parameters under strict management and a 10-day rotation cycle. This model effectively supports the treatment of acute hemorrhage and holds promise for integration into the future wartime blood supply system of the military.

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

Objective Genotypes(G)1,3,and 5 of the Japanese encephalitis virus(JEV)have been isolated in China,but the dominant genotype circulating in Chinese coastal areas remains unknown.We searched for G5 JEV-infected cases and attempted to elucidate which JEV genotype was most closely related to human Japanese encephalitis(JE)in the coastal provinces of China. Methods In this study,we collected serum specimens from patients with JE in three coastal provinces of China(Guangdong,Zhejiang,and Shandong)from 2018 to 2020 and conducted JEV cross-neutralization tests against G1,G3,and G5. Results Acute serum specimens from clinically reported JE cases were obtained for laboratory confirmation from hospitals in Shandong(92 patients),Zhejiang(192 patients),and Guangdong(77 patients),China,from 2018 to 2020.Seventy of the 361 serum specimens were laboratory-confirmed to be infected with JEV.Two cases were confirmed to be infected with G1 JEV,32 with G3 JEV,and two with G5 JEV. Conclusion G3 was the primary infection genotype among JE cases with a definite infection genotype,and the infection caused by G5 JEV was confirmed serologically in China.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.

Objective To investigate the effect of preoperative oral midazolam on postoperative de-lirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.Methods Eighty elderly patients undergoing laparoscopic surgery for radical resection of colorectal cancer,32 males and 48 females,aged 65-79 years,BMI 21-27 kg/m2,ASA physical status Ⅱ or Ⅲ,the state-trait anxiety inventory(STAI-S)≥38 scores at admission were selected.Patients were divided into two groups using random number method:control group and midazolam group,40 patients in each group.The midazolam group were administrated midazolam 7.5 mg per night till one day before surgery,while the placebo was administrated in the control group.The incidence of delirium 3 days after surgery and the STAI-S scores of one day before surgery were evaluated.The HR and MAP at entry,30 minutes after an-esthesia induction,1 hour,2 hours after anesthesia induction,and 30 minutes after extubation were recor-ded.The total dose of propofol,remifentanil and dexmedetomidine and the using rate of metaraminol were recorded.The visual analog scale scores 30 minutes after extubation,24 and 72 hours after surgery,the u-sing rate of tramadol,and the extubation time were recorded.Results Compared with the control group,the STAI-S scores of one day before surgery,and the incidence of postoperative delirium,the rate of using metaraminol,the VAS scores 30 minutes after extubation and 24 hours after surgery,the rate using of tram-adol were significantly decreased in the midazolam group(P＜0.05).There were no significant differences in total dose of propofol,remifentanil,and dexmedetomidine,extubation time between the two groups.Conclusion Preoperative oral midazolam can effectively reduce the incidence of postoperative delirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.

Objective:To explore the relationship between the expressions of P21,P27 and proliferating cell nuclear antigen(PCNA)in glomerular mesangial tissue and poor renal prognosis in patients with immunoglobulin A(IgA)nephropathy.Methods:A total of 145 patients with IgA nephropathy treated in Xiaogan Central Hospital from April 2017 to August 2019 were selected as the research object.The expressions of P21,P27 and PCNA in glomerular mesangial tissue were detected by immunohistochemistry.All patients were followed up for 24 months,and the prognosis were counted.The expressions of P21,P27 and PCNA in glomerular mesangial tissue of patients with different prognosis were compared and the influencing factors of poor prognosis in patients with IgA nephropathy were analyzed by Logistic regression analysis.Results:The expression rates of P21,P27 and PCNA positive cells in glomerular mesangial tissue of patients with IgA nephropathy were(38.69±6.83)%,(55.94±8.08)%,(33.47±5.72)%,respectively.The incidence rete of poor prognosis in patients with IgA nephropathy was 17.24%,and the expression rates of P21 and PCNA positive cells in glomerular mesangial tissue of patients with poor prognosis were higher than those in good prognosis group(P<0.05),while the expression rate of P27 positive cells was lower than that in good prognosis group(P<0.05).Logistic multiple regression analysis showed that elevated diastolic blood pressure,increased 24 h proteinuria,mesangial cell proliferation,segmental glomerulosclerosis,renal tubular atrophy/interstitial fibrosis,crescentic body,increased expression rates of P21 and PCNA positive cells and decreased expression rate of P27 positive cells were all risk factors affecting the poor prognosis of patients with IgA nephropathy(P<0.05).Conclusion:There are positive expressions of P21,P27 and PCNA in glomerular mesangial tissue of IgA nephropathy.The expression rates of P21 and PCNA positive cells in glomerular mesangial tissue of of patients with poor prognosis of IgA nephropathy are higher than those with good prognosis,while the expression rate of P27 protein positive cells is lower than those with good prognosis,which are risk factors for poor prognosis of patients with IgA nephropathy.