The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

Objective:To compare the efficacy of 68Ga-Pentixafor PET/CT in guiding surgical treatment of primary aldosteronism (PA) with that of adrenal venous sampling (AVS), and to explore its value in the diagnosis and management of PA. Methods:A total of 83 patients (62 males, 21 females, age (48.5±10.4) years) who received unilateral adrenal PA based on 68Ga-Pentixafor PET/CT or AVS results at the Department of Urological Surgery of Peking Union Medical College Hospital from January 1, 2021 to May 31, 2023 were retrospectively enrolled. Clinical data of patients were collected and the postoperative benefit rates guided by the two examination methods were compared according to the international multi-center PA surgical outcome standard. Quantitative indexes in 68Ga-Pentixafor PET/CT (SUV max, ratio of lesion SUV max to normal-adrenal-tissue SUV mean (LAR), ratio of lesion SUV max to normal-liver SUV mean (LLR), and ratio of lesion SUV max to contralateral SUV max (LCR)) were obtained for comparative analysis in patients with different surgical outcomes. Mann-Whitney U test, χ2 test or Fisher′s exact test were used to analyze the data. Results:Among 83 patients, 35 underwent AVS-guided surgery and 48 underwent 68Ga-Pentixafor PET/CT-guided surgery, with no significant difference of surgical benefit rates (85.7%(30/35) vs 85.4%(41/48); χ2=0.01, P=0.970). There was no significant difference of surgical benefit rates between 2 methods in such subgroups: <35 years (2/3 vs 7/8), ≥35 years (87.5%(28/32) vs 85.0%(34/40)), males (85.2%(23/27) vs 88.6%(31/35), females (7/8 vs 10/13), patients with unilateral lesions indicated by CT results (13/15 vs 83.9%(26/31)), patients with bilateral lesions indicated by CT results (85.0%(17/20) vs 15/17) (all χ2<0.01, all P>0.05). In PET/CT group, the LCR of patients who benefited from surgery ( n=41) was higher than that of patients who did not benefit ( n=7; 3.19(2.24, 4.90) vs 1.89(1.59, 2.88); Z=-2.09, P=0.036), and other quantitative indicators also tended to be higher. Conclusions:The positive results of 68Ga-Pentixafor PET/CT and AVS have the same clinical value in guiding PA patients to receive unilateral adrenal surgery. 68Ga-Pentixafor PET/CT is expected to become a non-invasive examination method to guide the treatment decisions in PA patients.

In the latest fifth edition of the World Health Organization（WHO）classification of endocrine and neuroendocrine tumors released in 2022，updates were made to the categorization of adrenocortical nodular diseases. The new term “sporadic nodular adrenocortical disease” has replaced “adrenocortical nodular hyperplasia” to describe incidentally detected，non-functional benign adrenocortical nodules with the diameter < 1 cm. This revised nomenclature reflects a deeper understanding of the nature of such conditions，emphasizing a shift from "hyperplastic changes" to the recognition of these nodules as independent monoclonal proliferative lesions. Sporadic nodular adrenocortical disease typically does not require surgical intervention and is managed primarily through outpatient follow-up and monitoring. Clinicians are advised to conduct systematic and standardized evaluations for patients presenting with adrenocortical nodules to avoid unnecessary surgeries or overtreatment. This article provides a concise commentary on the clinical implications of the conceptual updates to sporadic nodular adrenocortical disease outlined in the new WHO classification.

Objective:To explore the value of metanephrine, normetanephrine, and some steroid hormones in the assessment of adrenal venous sampling (AVS).Methods:This retrospective study enrolled 101 patients with primary aldosteronism who underwent AVS at Peking Union Medical College Hospital between June 1, 2021, and October 1, 2024. Multiple hormones, including aldosterone, cortisol, metanephrine, normetanephrine and steroid hormone profiles, were measured in samples from the inferior vena cava and bilateral adrenal veins during AVS. Selectivity index and lateralization index were calculated based on the levels of different hormones to determine successful AVS cannulation (selectivity index≥2) and aldosterone hypersecretion lateralization (lateralization index≥2). Patients who underwent unilateral adrenalectomy were followed for at least 6 months. Clinical and biochemical outcomes were assessed according to the Primary Aldosteronism Surgical Outcome (PASO) criteria, with biochemical remission defined as achieving complete or partial biochemical remission postoperatively. The efficacy of different hormones relative to cortisol for calculating selectivity index and lateralization index was evaluated for subtype classification.Results:The age at diagnosis of the enrolled patients was (50.5±9.6) years, including 77 males. Regarding the selectivity index, five hormones including metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone, and dehydroepiandrosterone demonstrated significantly higher selectivity index compared to cortisol (all P<0.05). Based on the cortisol-derived selectivity index, AVS cannulation was unsuccessful in 8 patients; using the five indices, unsuccessful cannulation occurred in 2, 2, 3, 4, and 5 patients, respectively. Based on postoperative follow-up, 55 patients were identified as having unilateral surgically relievable primary aldosteronism. In identifying these patients, the performance of metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone, and dehydroepiandrosterone was non-inferior to cortisol, correctly identifying 95% (52/55), 93% (51/55), 91% (50/55), 87% (48/55), and 89% (49/55) of cases, respectively. However, among these patients, there were no statistically significant differences in the success rate of intubation in AVS and the ability to identify patients with unilateral primary aldosteronism between the five indicators and cortisol (all P>0.05). Using cortisol-based lateralization as the reference standard, androstenedione and dehydroepiandrosterone both achieved an accuracy of 90% (84/93) for determining the lateralized side, while 17α-hydroxypregnenolone, normetanephrine, and metanephrine achieved accuracies of 89% (83/93), 81% (74/93), and 80% (73/93), respectively. Conclusion:Metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone and dehydroepiandrosterone could increase the success rate of intubation in AVS, with a high ability to identify patients with unilateral primary aldosteronism, and are expected to replace cortisol as new indicators of AVS.

Urothelial carcinoma (UC), including bladder urothelial carcinoma and upper tract urothelial carcinoma (UTUC), is the most common malignant tumor in the urinary system. Traditional cell line models fall short in simulating its tumor microenvironment and in vivo behavior. Patient-derived organoid (PDO) models offer a new way to overcome these shortcomings. This paper reviews the construction techniques of PDO models in UC, their biological simulation capabilities, and their applications in preclinical research. It also analyzes the technical limitations of these models. PDO models can retain the histological, genomic, and transcriptomic features of the parent tumor and accurately simulate the tumor microenvironment and biological behavior. They have been widely used in bladder cancer research, providing a precise platform for drug screening, personalized treatment, and immunotherapy evaluation. However, their use in UTUC research is still in its infancy. In the future, through technological optimization, PDO models are expected to enhance their value in UC research, advancing precision medicine research and clinical translation.

The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

Objective:To compare the efficacy of 68Ga-Pentixafor PET/CT in guiding surgical treatment of primary aldosteronism (PA) with that of adrenal venous sampling (AVS), and to explore its value in the diagnosis and management of PA. Methods:A total of 83 patients (62 males, 21 females, age (48.5±10.4) years) who received unilateral adrenal PA based on 68Ga-Pentixafor PET/CT or AVS results at the Department of Urological Surgery of Peking Union Medical College Hospital from January 1, 2021 to May 31, 2023 were retrospectively enrolled. Clinical data of patients were collected and the postoperative benefit rates guided by the two examination methods were compared according to the international multi-center PA surgical outcome standard. Quantitative indexes in 68Ga-Pentixafor PET/CT (SUV max, ratio of lesion SUV max to normal-adrenal-tissue SUV mean (LAR), ratio of lesion SUV max to normal-liver SUV mean (LLR), and ratio of lesion SUV max to contralateral SUV max (LCR)) were obtained for comparative analysis in patients with different surgical outcomes. Mann-Whitney U test, χ2 test or Fisher′s exact test were used to analyze the data. Results:Among 83 patients, 35 underwent AVS-guided surgery and 48 underwent 68Ga-Pentixafor PET/CT-guided surgery, with no significant difference of surgical benefit rates (85.7%(30/35) vs 85.4%(41/48); χ2=0.01, P=0.970). There was no significant difference of surgical benefit rates between 2 methods in such subgroups: <35 years (2/3 vs 7/8), ≥35 years (87.5%(28/32) vs 85.0%(34/40)), males (85.2%(23/27) vs 88.6%(31/35), females (7/8 vs 10/13), patients with unilateral lesions indicated by CT results (13/15 vs 83.9%(26/31)), patients with bilateral lesions indicated by CT results (85.0%(17/20) vs 15/17) (all χ2<0.01, all P>0.05). In PET/CT group, the LCR of patients who benefited from surgery ( n=41) was higher than that of patients who did not benefit ( n=7; 3.19(2.24, 4.90) vs 1.89(1.59, 2.88); Z=-2.09, P=0.036), and other quantitative indicators also tended to be higher. Conclusions:The positive results of 68Ga-Pentixafor PET/CT and AVS have the same clinical value in guiding PA patients to receive unilateral adrenal surgery. 68Ga-Pentixafor PET/CT is expected to become a non-invasive examination method to guide the treatment decisions in PA patients.

In the latest fifth edition of the World Health Organization（WHO）classification of endocrine and neuroendocrine tumors released in 2022，updates were made to the categorization of adrenocortical nodular diseases. The new term “sporadic nodular adrenocortical disease” has replaced “adrenocortical nodular hyperplasia” to describe incidentally detected，non-functional benign adrenocortical nodules with the diameter < 1 cm. This revised nomenclature reflects a deeper understanding of the nature of such conditions，emphasizing a shift from "hyperplastic changes" to the recognition of these nodules as independent monoclonal proliferative lesions. Sporadic nodular adrenocortical disease typically does not require surgical intervention and is managed primarily through outpatient follow-up and monitoring. Clinicians are advised to conduct systematic and standardized evaluations for patients presenting with adrenocortical nodules to avoid unnecessary surgeries or overtreatment. This article provides a concise commentary on the clinical implications of the conceptual updates to sporadic nodular adrenocortical disease outlined in the new WHO classification.

Urothelial carcinoma (UC), including bladder urothelial carcinoma and upper tract urothelial carcinoma (UTUC), is the most common malignant tumor in the urinary system. Traditional cell line models fall short in simulating its tumor microenvironment and in vivo behavior. Patient-derived organoid (PDO) models offer a new way to overcome these shortcomings. This paper reviews the construction techniques of PDO models in UC, their biological simulation capabilities, and their applications in preclinical research. It also analyzes the technical limitations of these models. PDO models can retain the histological, genomic, and transcriptomic features of the parent tumor and accurately simulate the tumor microenvironment and biological behavior. They have been widely used in bladder cancer research, providing a precise platform for drug screening, personalized treatment, and immunotherapy evaluation. However, their use in UTUC research is still in its infancy. In the future, through technological optimization, PDO models are expected to enhance their value in UC research, advancing precision medicine research and clinical translation.

Objective:To explore the value of metanephrine, normetanephrine, and some steroid hormones in the assessment of adrenal venous sampling (AVS).Methods:This retrospective study enrolled 101 patients with primary aldosteronism who underwent AVS at Peking Union Medical College Hospital between June 1, 2021, and October 1, 2024. Multiple hormones, including aldosterone, cortisol, metanephrine, normetanephrine and steroid hormone profiles, were measured in samples from the inferior vena cava and bilateral adrenal veins during AVS. Selectivity index and lateralization index were calculated based on the levels of different hormones to determine successful AVS cannulation (selectivity index≥2) and aldosterone hypersecretion lateralization (lateralization index≥2). Patients who underwent unilateral adrenalectomy were followed for at least 6 months. Clinical and biochemical outcomes were assessed according to the Primary Aldosteronism Surgical Outcome (PASO) criteria, with biochemical remission defined as achieving complete or partial biochemical remission postoperatively. The efficacy of different hormones relative to cortisol for calculating selectivity index and lateralization index was evaluated for subtype classification.Results:The age at diagnosis of the enrolled patients was (50.5±9.6) years, including 77 males. Regarding the selectivity index, five hormones including metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone, and dehydroepiandrosterone demonstrated significantly higher selectivity index compared to cortisol (all P<0.05). Based on the cortisol-derived selectivity index, AVS cannulation was unsuccessful in 8 patients; using the five indices, unsuccessful cannulation occurred in 2, 2, 3, 4, and 5 patients, respectively. Based on postoperative follow-up, 55 patients were identified as having unilateral surgically relievable primary aldosteronism. In identifying these patients, the performance of metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone, and dehydroepiandrosterone was non-inferior to cortisol, correctly identifying 95% (52/55), 93% (51/55), 91% (50/55), 87% (48/55), and 89% (49/55) of cases, respectively. However, among these patients, there were no statistically significant differences in the success rate of intubation in AVS and the ability to identify patients with unilateral primary aldosteronism between the five indicators and cortisol (all P>0.05). Using cortisol-based lateralization as the reference standard, androstenedione and dehydroepiandrosterone both achieved an accuracy of 90% (84/93) for determining the lateralized side, while 17α-hydroxypregnenolone, normetanephrine, and metanephrine achieved accuracies of 89% (83/93), 81% (74/93), and 80% (73/93), respectively. Conclusion:Metanephrine, normetanephrine, androstenedione, 17α-hydroxypregnenolone and dehydroepiandrosterone could increase the success rate of intubation in AVS, with a high ability to identify patients with unilateral primary aldosteronism, and are expected to replace cortisol as new indicators of AVS.