1.Advances in the clinical management of primary hyperaldosteronism
Huasheng LIAO ; Yizhao WU ; Cai DENG ; Zijian AO ; Lichao ZHANG
Journal of Modern Urology 2025;30(9):797-802
Primary hyperaldosteronism(PHA),the most common cause of secondary hypertension,can lead to cardiovascular and renal damage events.Early diagnosis and accurate typing of PHA is crucial to the choice of treatment options,and its diagnostic modalities usually include prone test screening,captopril test characterization and adrenal vein blood sampling for localization and typing diagnosis.However,with the development of imaging,molecular biology and morphology technologies,the use of nuclear medicine PET-CT,genetic testing and pathological diagnostic methods can also be used to accurately typify PHA.New technologies such as adrenal artery embolization and adrenal radiofrequency ablation are also increasingly being used in the treatment of PHA,which have the advantages of shorter and less costly surgery,fewer postoperative complications,and quicker recovery.This article provides a review of the developments in the screening,diagnosis and treatment of PHA with a view to informing clinical practice.
2.Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application
Meizhen OU ; Yongfeng LI ; Sha WEN ; Zhouxiang LIAO ; Xuejing HUANG ; Lichao YANG ; Min HE
Laboratory Animal and Comparative Medicine 2025;45(5):611-622
Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90%,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.
3.Advances in the clinical management of primary hyperaldosteronism
Huasheng LIAO ; Yizhao WU ; Cai DENG ; Zijian AO ; Lichao ZHANG
Journal of Modern Urology 2025;30(9):797-802
Primary hyperaldosteronism(PHA),the most common cause of secondary hypertension,can lead to cardiovascular and renal damage events.Early diagnosis and accurate typing of PHA is crucial to the choice of treatment options,and its diagnostic modalities usually include prone test screening,captopril test characterization and adrenal vein blood sampling for localization and typing diagnosis.However,with the development of imaging,molecular biology and morphology technologies,the use of nuclear medicine PET-CT,genetic testing and pathological diagnostic methods can also be used to accurately typify PHA.New technologies such as adrenal artery embolization and adrenal radiofrequency ablation are also increasingly being used in the treatment of PHA,which have the advantages of shorter and less costly surgery,fewer postoperative complications,and quicker recovery.This article provides a review of the developments in the screening,diagnosis and treatment of PHA with a view to informing clinical practice.
4.Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application
Meizhen OU ; Yongfeng LI ; Sha WEN ; Zhouxiang LIAO ; Xuejing HUANG ; Lichao YANG ; Min HE
Laboratory Animal and Comparative Medicine 2025;45(5):611-622
Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90%,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.
5.Establishment and validation of a prediction model of the right adrenal dominant secretory side in patients with primary hyperaldosteronism
Huasheng LIAO ; Yizhao WU ; Cai DENG ; Lichao ZHANG
Journal of Chinese Physician 2025;27(3):382-387
Objective:To explore the independent risk factors for predicting the right adrenal dominant secretory side in patients with primary hyperaldosteronism (PHA), so as to construct and verify the prediction model of the dominant secretory side, and provide evidence for the diagnosis and treatment of PHA patients.Methods:Clinical data of 82 PHA patients undergoing adrenal vein sampling (AVS) admitted to the Second Affiliated Hospital of Guangzhou Medical University from January 2015 to July 2023 were retrospectively analyzed, and divided into the training set (58 cases) and the verification set (24 cases) according to the ratio of 7∶3. Single factor and multiple factor logistic regression analysis were used to screen out the risk factors for predicting the right adrenal dominant secretory side, and then combined with the risk factors to build the prediction model of the dominant secretory side. The predictive efficiency and clinical applicability of the predominance secretory nomogram prediction model were verified by drawing receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis (DCA curve).Results:(1) Univariate and multivariate logistic analysis showed that the concentration of aldosterone (ALD2) in inferior vena cava and the results of adrenal CT imaging suggested that the right adrenal unilateral nodule (RJJ) were independent risk factors for the occurrence of right adrenal dominant secretion in PHA patients undergoing AVS (all P<0.05). (2) The area under ROC curve (AUC) corresponding to ALD2 variable was 0.898(95% CI: 0.735 7-0.986 1), and the sensitivity and specificity were 95.2% and 81.1%, respectively. The corresponding AUC of RJJ variable was 0.750(95% CI: 0.683 1-0.976 3), and the sensitivity and specificity were 90.5% and 86.7%, respectively. The AUC of the two variables in the verification set were 0.926 and 0.778, respectively. (3) Calibration curve and DCA curve showed that the prediction model had good consistency and clinical applicability, and can produce good clinical benefits. Conclusions:ALD2 and adrenal CT results suggest that RJJ is an independent risk factor for the prediction of the right adrenal dominant secretory side, and the prediction model based on these risk factors has good predictive efficacy and clinical applicability.
6.Establishment and validation of a prediction model of the right adrenal dominant secretory side in patients with primary hyperaldosteronism
Huasheng LIAO ; Yizhao WU ; Cai DENG ; Lichao ZHANG
Journal of Chinese Physician 2025;27(3):382-387
Objective:To explore the independent risk factors for predicting the right adrenal dominant secretory side in patients with primary hyperaldosteronism (PHA), so as to construct and verify the prediction model of the dominant secretory side, and provide evidence for the diagnosis and treatment of PHA patients.Methods:Clinical data of 82 PHA patients undergoing adrenal vein sampling (AVS) admitted to the Second Affiliated Hospital of Guangzhou Medical University from January 2015 to July 2023 were retrospectively analyzed, and divided into the training set (58 cases) and the verification set (24 cases) according to the ratio of 7∶3. Single factor and multiple factor logistic regression analysis were used to screen out the risk factors for predicting the right adrenal dominant secretory side, and then combined with the risk factors to build the prediction model of the dominant secretory side. The predictive efficiency and clinical applicability of the predominance secretory nomogram prediction model were verified by drawing receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis (DCA curve).Results:(1) Univariate and multivariate logistic analysis showed that the concentration of aldosterone (ALD2) in inferior vena cava and the results of adrenal CT imaging suggested that the right adrenal unilateral nodule (RJJ) were independent risk factors for the occurrence of right adrenal dominant secretion in PHA patients undergoing AVS (all P<0.05). (2) The area under ROC curve (AUC) corresponding to ALD2 variable was 0.898(95% CI: 0.735 7-0.986 1), and the sensitivity and specificity were 95.2% and 81.1%, respectively. The corresponding AUC of RJJ variable was 0.750(95% CI: 0.683 1-0.976 3), and the sensitivity and specificity were 90.5% and 86.7%, respectively. The AUC of the two variables in the verification set were 0.926 and 0.778, respectively. (3) Calibration curve and DCA curve showed that the prediction model had good consistency and clinical applicability, and can produce good clinical benefits. Conclusions:ALD2 and adrenal CT results suggest that RJJ is an independent risk factor for the prediction of the right adrenal dominant secretory side, and the prediction model based on these risk factors has good predictive efficacy and clinical applicability.
7.Application of micro-lecture system based on ASSURE model in clinical practice teaching of dermatology
Lichao LIAO ; Juan WANG ; Qiong LIU ; Ruixue CHANG ; Siping ZHANG
Chinese Journal of Medical Education Research 2021;20(10):1165-1168
Objective:To explore the application of micro-lecture system based on ASSURE model in clinical practice teaching of dermatology.Methods:Sixty clinical medicine undergraduate intern doctors in Batch 2014 of Anhui Medical University were randomized into 2 groups, with thirty interns in both experimental group and control group. Micro-lecture system based on ASSURE model teaching method was adopted in experimental group, while traditional teaching method was adopted in control group. At the end of practice, theory test scores and medical record writing was used to evaluate the clinical ability of interns in two groups, and questionnaire survey was conducted to evaluate the satisfaction of interns with the teaching method of micro-lecture system based on ASSURE model. SPSS 16.0 was used for t test. Results:①Theory test scores were higher in experimental group than in control group, with statistically significant differences between the two groups ( P<0.05); ②The scores of history of illness, physical examination, treatment plan and the medical record writing were higher in experimental group than in control group ( P<0.05); ③At the end of internship, the questionnaire showed that the interns in the experimental group were more recognized for this new way of teaching. Conclusion:Micro-lecture system based on ASSURE model teaching method has achieved better results in the teaching of dermatology, which is worthy of future research and promotion.
8.Supratentorial primitive neutoectodermal tumors in adults:imaging findings and analysis on misdiagnosis
Shukun LIAO ; Xiaodan LI ; Liuji GUO ; Lichao MA ; Jie DING ; Yikai XU ; Yuankui WU
Journal of Practical Radiology 2018;34(2):176-179
Objective To investigate the CT and MRI features of supratentorial primitive neuroectodermal tumors (sPNET)in adults,and to analyze the reasons of misdiagnosis.Methods The CT and MRI features of 1 5 patients with sPNET confirmed by pathology were analyzed retrospective.13 of 15 patients were underwent plain and contrast enhanced MRI,4 patients were also performed CT scan and the other 2 patients were underwent CT plain scan only.Results Nine of 1 5 lesions were located in the temporal and/or frontal lobe,with clear margin in 12 lesions,and mild to moderate peritumoral was noted in most cases.The solid portion of the lesions were hyperdense on CT,isointense or slightly hypointense on T1WI,and isointense or hyperintense on T2WI compared to the gray matter.Calcification (2/6),necrosis or cystic degeneration(1 3/1 5),hemorrhage (8/1 5)and flow void signal (1 1/1 3)were seen.Twelve lesions showed significantly heterogeneous enhancement and 7 lesions showed irregular ring-enhancement.The solid parts in 5 lesions showed hyperintensity on diffusion-weighted imaging. Preoperatively,the lesions were misdiagnosed as glioma in 10 cases,ependymoma in 2 cases,meningioma in 1 case and germ cell tumor in 1 case, respectively;and 1 case was not diagnosed definitely.Conclusion Supratentorial PNET in adults has characteristic CT and MRI features,and the main reasons accounting for misdiagnosis are its extraordinary low incidence and that radiologists do no master its imaging features.
9.Clinical analysis of 18 cases of neurosyphilis
Faxing JIANG ; Bai HU ; Qiqiang TANG ; Zhenglong ZHAO ; Aili WU ; Siping ZHANG ; Lichao LIAO ; Jinli LIU ; Hailin ZHOU ; Qianqiu WANG
Chinese Journal of Dermatology 2010;43(5):301-304
Objective To analyze the clinical features, diagnosis, treatment and prognosis of neurosyphilis. Methods Clinical data on and laboratory findings in 18 cases with neurosyphilis collected in the Affiliated Provincial Hospital, Anhui Medical University from 2006 to 2008 were retrospectively studied.Results Among the 18 patients, 3 sufferred from asymptomatic neurosyphilis, 1 from meningeal syphilis, 7 from meningovascular syphilis, 5 from paralytic dementia, and 2 from intracranial space-occupation. Toluidine red unheated serum reagin test (TRUST) and Treponema pallidum particle agglutination test (TPPA) of sera were positive in all the patients; cerebrospinal fluid (CSF) TRUST was positive in 16 patients, and CSF TPPA in all patients. An increase was observed in CSF leukocyte count in 7 patients and in CSF protein in 13 patients.The findings on cerebral magnetic resonance imaging (MRI) mainly included demyelination, brain atrophy,cerebral infarction, etc. All the patients, except 2 with a TRUST titer of 1:4, experienced a 4-fold decrease in TRUST titer within a 3-month follow up. Clinical symptoms of neurosyphilis improved in all patients except 1 with paralytic dementia. Conclusions The diversity of clinical manifestations usually leads to the misdiagnosis of neurosyphilis, which should be diagnosed based on comprehensive analysis of clinical characteristics as well as laboratory and imaging findings. Early diagnosis and treatment are beneficial to its prognosis.
10.Application of immunohistochemistry in adrenal cortical neoplasm and hyperplasia
Lichao ZHANG ; Weilie HU ; Qiuling LIAO ; Xiaodong CHEN
Journal of Medical Postgraduates 2003;0(08):-
Objective:To study the characteristics of chromogranin(CgA),synaptophysin(SYN) and neuron-specific enolase(NSE) in the cortex of adrenal gland.Methods:Immunohistochemical technique was used to detect the expressions of CgA,SYN and NSE in 56 routinely processed tissue specimens from human adrenal cortical neoplasm and hyperplasia.Results: CgA immunoreactivity was regularly detected in the cortex of adrenal neoplasm,but not in the hyperplasia and normal adrenal gland.The immunoreactive materials appeared in the cytoplasm and in the form of vacuole or grains.Adrenal cortical neoplasm and hyperplasia showed NSE positive cells in the cortex,but not in the normal adrenal gland.SYN positive materials were shown in the cortex of hyperplasia and adrenocorticoadenoma,but not in the adrenocorticoadenocarcinoma and normal adrenal gland.Conclusion: The distributive characteristics of CgA,SYN and NSE in the adrenal cortex help the differential diagnosis of adrenal hyperplasia.The adrenal cortex is closely connected with medulla.Besides the classical hypothalamus-pituitary-adrenal axis(HPAA) and rennin-angiotensin-aldosterone system(RAAS),the adrenal cortex and medulla are mutually regulated on the basis of anatomy and endocrinology.

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