ObjectiveTo systematically summarize the molecular mechanisms, multimodal localization, and genetic stratification-based management strategies of primary hyperparathyroidism (PHPT), and to offer theoretical references for the precision diagnosis and treatment of PHPT. MethodRelevant literature on the molecular mechanisms, multimodal localization, and genetic stratification-based treatment of PHPT was retrieved from domestic and international databases and systematically reviewed. ResultsThe pathogenesis of PHPT involves cell cycle dysregulation driven by MEN1 and CCND1, impaired negative feedback of calcium homeostasis resulting from mutations in the CaSR (calcium-sensing receptor) and GNA11 (guanine nucleotide-binding protein subunit alpha-11), and the risks of malignant transformation associated with CDC73 mutations. Combined ultrasonography and radionuclide imaging can significantly enhance preoperative localization accuracy. Four-dimensional computed tomography (CT) yields high localization success rates for solitary parathyroid adenomas, whereas 18F-fluorocholine PET-CT exhibits excellent diagnostic performance in patients with negative 99mTc-MIBI (technetium-99m methoxyisobutylisonitrile) findings or multiglandular disease. Germline genetic testing is recommended for patients with early-onset PHPT, recurrent or multiglandular parathyroid disease, or suspected parathyroid carcinoma to guide the extent of surgical resection and avoid unnecessary bilateral neck exploration. Minimally invasive parathyroidectomy is the gold standard for solitary parathyroid adenomas. Ablation therapy is a viable option for patients who are unfit for surgery, whereas pharmacotherapy should be tailored according to whether hypercalcemia or skeletal disease is the predominant feature. ConclusionsPrecision management of PHPT requires the integration of four core pillars: biochemical characterization, molecular mechanisms, multimodal imaging, and genetic stratification. A stratified diagnostic framework coupled with a tiered clinical pathway can overcome the traditional “one-size-fits-all” empirical treatment model and establish a standardized closed-loop system of “screening–referral–follow-up”, thereby improving overall diagnostic yield and therapeutic outcomes.
Objective To compare the characteristic differences between apoptotic extracellular vesicle (ApoEV) and non-apoptotic cell-derived extracellular vesicle (NCEV) from bone marrow mesenchymal stem cells (BMSCs), and to explore their therapeutic effects on inflammatory macrophages and cisplatin-induced acute kidney injury (AKI) in mice. Methods Staurosporine (STS) was used to induce apoptosis in BMSCs, and the apoptotic model was verified by cell morphology observation, Annexin V/propidium iodide double-staining flow cytometry, and Western blot. NCEV and ApoEV were isolated via differential centrifugation combined with ultracentrifugation, and characterized using transmission electron microscopy, nanoparticle tracking analysis, and Western blot. The particle yield and protein yield of extracellular vesicles (EV) per unit donor cell were compared between the two groups. EVs were labeled with DiD, and the uptake of EV by RAW264.7 macrophages was detected using laser scanning confocal microscopy and flow cytometry. Lipopolysaccharide (LPS) was applied to stimulate RAW264.7 cells to establish an in vitro inflammatory cell model. The mRNA expression levels of interleukin 10 (IL-10), arginase 1 (Arg-1), IL-1β, IL-6, IL-18, and tumor necrosis factor α (TNF-α) were determined by real-time fluorescence quantitative PCR. The intracellular reactive oxygen species (ROS) level was measured using 2’, 7’-dichlorodihydrofluorescein diacetate fluorescent probe with laser scanning confocal microscopy and flow cytometry. EVs were labeled with cyanine 7 N-hydroxysuccinimide ester to observe their distribution in major organs and renal tissues of normal mice. A cisplatin-induced AKI mouse model was established, followed by intervention with NCEV or ApoEV. Serum creatinine (Scr), blood urea nitrogen (BUN), renal histopathological injury, kidney injury molecule 1 (Kim-1), cell apoptosis, macrophage infiltration, and DNA damage-related indicators were detected. Based on previously published proteomic data of EV derived from mouse BMSCs, differentially expressed proteins were screened and gene ontology enrichment analysis was performed. Results After STS treatment, BMSCs exhibited typical apoptotic morphology with increased early and late apoptotic rates, alongside upregulated expression of Cleaved Caspase-3 and Bcl-2-associated X protein (Bax). Both NCEV and ApoEV presented spherical or quasi-spherical structures enclosed by a lipid bilayer membrane, with similar particle sizes and Zeta potentials. Bax and tumor susceptibility gene 101 protein were detected in both vesicles, while Cleaved Caspase-3 and Bax were detectable in ApoEV. The particle yield and protein yield of ApoEV were (5.626±0.302)-fold and (2.972±0.124)-fold higher than those of NCEV, respectively (P<0.05). Compared with NCEV, RAW264.7 cells displayed significantly elevated uptake ratio and mean fluorescence intensity for ApoEV (P<0.05). Under LPS stimulation, ApoEV markedly upregulated the mRNA expressions of IL-10 and Arg-1 to levels higher than those in the NCEV treatment group (P<0.05), whereas NCEV significantly suppressed the mRNA expressions of IL-1β and IL-6. Both NCEV and ApoEV reduced intracellular ROS levels, and flow cytometry revealed that the mean 2’, 7’-dichlorofluorescein fluorescence intensity in the ApoEV group was significantly lower than that in the NCEV group (P<0.05). Ex vivo organ imaging demonstrated that the strongest fluorescent signals of both types of EVs appeared in the liver, and obvious signals were also observed in the lung, spleen, and kidney. No significant difference in relative radiation efficiency of each organ was found between the two groups (P>0.05). In cisplatin-induced AKI mice, NCEV significantly reduced serum Scr and BUN levels, while ApoEV significantly decreased serum Scr levels. Both EV preparations alleviated renal tubular histopathological damage and lowered the levels of Kim-1, Bax, TUNEL-positive cells, F4/80-positive macrophages, and serine 139-phosphorylated histone H2AX. No significant intergroup differences in the above renal injury indicators were observed between the two EV intervention groups (P>0.05). Proteomic analysis identified 50 significantly upregulated proteins and 35 significantly downregulated proteins in ApoEV relative to NCEV. The upregulated proteins in ApoEV were mainly enriched in biological processes including extracellular matrix organization, classical complement pathway activation, proteolysis, and angiogenesis. Conclusion ApoEV possess higher production yield, enhanced macrophage uptake capacity and superior anti-inflammatory regulatory activity compared with NCEV. Both ApoEV and NCEV can alleviate renal injury in cisplatin-induced AKI, providing experimental evidence supporting the translational application of ApoEV for AKI treatment.
After more than 10 years of development and construction, the continuous renal replacement therapy subspecialty of the Department of Nephrology of West China Hospital of Sichuan University has made outstanding achievements in the aspect of continuing education. This article discusses and summarizes the current training measures for continuous renal replacement therapy subspecialized refresher physicians in the Department of Nephrology of West China Hospital of Sichuan University, and introduces the training goals, training measures and training results. The purpose is to provide a summary of experience for the continuing education of continuous renal replacement therapy subspecialized refresher physicians, provide a reference for hospitals that plan to develop continuous renal replacement therapy subspecialized refresher physicians training, and assists in promoting the development of continuous renal replacement therapy subspecialized continuing education in China.
To investigate the microsurgical management of cranionasal tumors and the method of the reconstruction of the skull base. Methods From June 2005 to October 2007, 20 patients with cranionasal tumor were treated. There were 10 males and 10 females, aged between 13 and 77 years (median 49 years). The disease course was 2 months to 13 years.The cranionasal tumors, proved by MRI and CT scans, located in the anterior skull base, paranasal sinus, nasal and/or orbit cavity. And their cl inical presentations were l isted as follows: dysosphresia in 14 patients, headache in 11 patients, nasal obstruction in 9 patients, epistaxis in 8 patients, visual disorder in 4 patients, exophthalmos in 4 patients and conscious disturbance in 2 patients. All 20 patients underwent transbasal surgery combined with transnasal surgery, and tumors were resected by one-stage operation. The skull base was reconstructed by surgical technique “Pull Down Sandwich” with pedicle periosteum flap. Results Tumors were resected by one-stage operation, and the anterior skull bases were reconstructed. Pathological examination showed 8 cases of mal ignant tumors and 12 cases of benign tumors. The total surgical excision was complete in 16 patients, and 4 patients with subtotal excision. There was no operative death. Eighteen patients were followed up 3 months to 2 years and 6 months. Transient cerebrospinal fluid rhinorrhea was found in 2 cases which were cured by lumbar drainage. And recurrence of tumor was observed in 5 patients 3 months to 2 years after operation. Conclusion Microsurgical operation via subfrontal approach assisted bytransnasal endoscopy is an effective method in management of cranionasal tumors, with the advantages of econstruction of the skull base with pedicle periosteum flap or “Pull Down Sandwich” and low compl ication rate.
Objective To study the protective effects of bone marrow mesenchymal stem cells (BMSCs) of rhesus monkeys on porcine islets from hypoxia/reoxygenation (H/R)-induced injury. Methods BMSCs were isolated and cultured from the marrow of 5 adult rhesus monkeys (weighing, 6-10 kg) by adherent monocytes. Islets were isolated and purified from the pancreas of 5 neonatal porcine (3-5 days old) by collagenase V digestion method, and were cultured with or without BMSCs, and exposed to hypoxia (1%O2) for 12 hours and reoxygenation for 24 or 48 hours, respectively. The experiment was divided into 4 groups: normal islet group (group A), normal islet + BMSCs group (Group B), H/R islet group (group C), and H/R islet + BMSCs group (group D). The survival rate of islets was calculated by fluorescein diacetate/propidium iodide (PI) staining. The viability of the islet cells was detected by cell counting kit 8. Apoptotic rate of islet cells was tested using Annexin V-FITC/PI labeling and flow cytometry. The stimulation index (SI) of islet function was analyzed by glucose-stimulated insulin secretion assay. Results The islet cell cluster of group C was more dispersed than that of groups A and B, and group C had more death cells; and the islet cell cluster of group D was more complete and the survival rate was higher than those of group C. The survival rate of islet was 90.2% ± 9.1%, 88.3% ± 5.9%, 52.3% ± 12.1%, and 71.4% ± 11.5% in groups A, B, C, and D respectively, it was significantly lower in groups C and D than in groups A and B (P lt; 0.05), but it was significantly higher in group D than in group C (P lt; 0.05). After coculture of BMSCs and islet at the ratio of 1 ∶ 10 and 1 ∶ 20 in group D, the viability of islet cells was significantly higher than that in group C (P lt; 0.05). The apoptotic rate was 27.1% ± 3.2%, 24.0% ± 1.0%, 64.3% ± 1.8%, and 46.2% ± 1.4% in groups A, B, C, and D respectively, it was significantly higher in groups C and D than that in groups A and B (P lt; 0.05), but it was significantly lower in group D than in group C (P lt; 0.05). There was no significant difference in SI between groups A and B at each time point (P gt; 0.05), but it was significantly lower in group C than in groups A and B (P lt; 0.05); and it was significantly higher in group D than in group C at 24 and 72 hours (P lt; 0.05). Conclusion BMSCs of rhesus monkeys can protect islet vitality and function from H/R-induced injury.
ObjectiveTo explore the optimal conditions of rat model of hyperuricemia (HUA) induced by different doses of potassium oxanate (PO) combined with adenine, and to provide reference for the treatment of HUA.MethodsMale Sprague-Dawley rats (220-240 g body weight) were divided into normal control group, potassium oxanate (1000, 1500 mg/kg) and adenine (0, 50, 100 mg/kg) combined model groups, with 8 rats in each group. After 5 weeks of intragastric administration, blood were collected from tail vein of rats every week, and serum uric acid, creatinine and blood urea nitrogen level were measured. At the 6th week, the changes of the pathological characteristics, expression of inflammatory and fibrosis-related factors in the kidneys were observed.ResultsIn the 1500 mg/kg potassium oxanate combined with 100 mg/kg adenine group, rats died after 2 weeks of molding, and the survival rate at the 6th week was 62.5%; but there was no significant difference between the other groups and the normal control group in survival rate (P>0.05). Compared with the normal group, the level of serum uric acid in each model group increased significantly after 1 week of molding (P<0.05), but recovered to the pre-model level after stopping intragastric administration in week 6. After 5 weeks, in model groups the levels of serum creatinine and blood urea nitrogen were higher than those in the normal control group; and the inflammation and fibrosis-related factors mRNA and protein expression of kidney tissue in model groups increased with the increase of ademine dose, and there was a significant difference in the PO 1 000 mg/kg with adenine 100 mg/kg group, PO 1 500 mg/kg with Adenine 50 mg/kg group compared to the normal control group (P<0.05). The results of renal anatomy and histology testing in rats showed that with the increased of the dosage of PO and adenine in the model groups, the increase of white deposition of renal medulla, tubulointerstitial fibrosis, and tubular epithelial cell necrosis was found, and the glomerular atrophy aggravated. Compared with the indexes in the normal control group, the expression levels of inflammation and fibrosis related genes and proteins in the 50 mg/kg adenine combined with 1 500 mg/kg PO group were higher, and inflammatory cell infiltration and fibrosis were observed, which was consistent with the clinical manifestation of hyperuricemia induced renal injury.ConclusionPO (1500 mg/kg) combined with adenine (50 mg/kg) can establish a stable hyperuricemic nephropathy model in rats.