1.Exploration of Rat Fetal Lung Tissue Fixation Methods
Liyu LIU ; Bo JI ; Xiaoxuan LIU ; Yang FANG ; Ling ZHANG ; Tingting GUO ; Ye QUAN ; Hewen LI ; Yitian LIU
Laboratory Animal and Comparative Medicine 2025;45(4):432-438
ObjectiveThis study explores the methods of lung tissue extraction and fixation required for pathological studies of fetal rats, based on the unique physiological structure of fetal rat lung tissue and existing lung tissue fixation techniques for adult rats. MethodsSix pregnant adult SD rats at 20.5 days of gestation were subjected to cesarean section to obtain fetal rats. Four healthy fetal rats with similar body weight, vital signs, and respiratory status were selected from each pregnant rat, and they were randomly divided into the following groups using a random number table: direct lung infiltration group, lung infiltration group after intratracheal infusion, whole-body infiltration group of fetal rats, and whole-body infiltration group after intratracheal infusion of fetal rats. To systematically compare and analyze the anatomical morphology under different fixation methods, lung tissues from four groups of fetal rats were harvested, perfused, and fixed, and the gross morphology of lung tissues in each group was observed. Paraffin sections were prepared and stained with Hematoxylin-Eosin (H&E). The histological morphology of the whole lung, alveoli, and bronchi was further examined under optical microscopy. ResultsIn the direct lung infiltration group, the hilar structures were unclear, lung lobation was indistinct, the shape was irregular, lung cavities were small, and alveoli and bronchi were shrunken. In the lung infiltration group after intratracheal infusion, the hilar structures were clear, lobation was pronounced, the shape was regular, lung cavities were large, and alveoli and bronchi were full. Both the whole-body infiltration group and whole-body infiltration group after intratracheal infusion of fetal rats exhibited visible lungs, hearts, skins, and other organs. The lung tissues of both groups showed obvious lobulation, irregular shape, and damage at the margins of lung lobes. In the whole-body infiltration group, the thoracic cavities of the fetus were flattened, lung cavities were small, and alveoli and bronchi were shrunken. In the whole-body infiltration group after intratracheal infusion of fetal rats, the fetal thoracic cavities were full, lung cavities were large, and alveoli and bronchi were relatively full. ConclusionThe lung infiltration after intratracheal infusion method for fetal rat lung tissue fixation outperforms direct lung infiltration, whole-body infiltration of fetal rats, and whole-body infiltration after intratracheal infusion of fetal rats in terms of preservation of the lung tissue's original morphology, paraffin sectioning, staining, and pathological observation and analysis. The embedding, sectioning, and staining processes are also simple and save consumables. Therefore, intratracheal infusion followed by lung infiltration method is recommended for fixation in histopathological observation of fetal rat lung tissue.
2.The Scientific Connotation of"Heart and Brain Co-Dominating the Mind"and Its Application in the Treatment of Insomnia by Acupuncture
Yuting ZHANG ; Shan QIN ; Xiaoqiu WANG ; Liyu LIN ; Mufeng LI ; Wenzhong WU ; Chengyong LIU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):181-188
The paper discusses the scientific connotation of"heart and brain co-dominating the mind",including the connotation of"heart and brain co-dominating the mind"in traditional Chinese medicine and the physiological and pathological relationship be-tween heart and brain in modern medicine.Based on the understanding of"heart and brain co-dominating the mind",this paper be-lieves that"restlessness of mind"is the core pathogenesis of insomnia,and regulating mind is the key to the acupuncture and moxibus-tion treatment of insomnia.The autonomic nervous system(ANS)is a bridge connecting the heart and brain,and is closely related to the central anatomy,neurotransmitters and physiological rhythms of sleep.The imbalance of ANS is highly consistent with the patho-genesis of"heart and brain restlessness"in traditional Chinese medicine.The Governor Vessel runs through the brain and the heart,and the"Governor Vessel regulating spirit acupuncture"can communicate the heart and brain,regulate ANS function,and thus im-prove sleep quality.
3.Comparison of electroacupuncture and dexamethasone on lung development impairment in offspring rats induced by prenatal nicotine exposure
Tingting GUO ; Bo JI ; Yang FANG ; Yitian LIU ; Hewen LI ; Xiaoxuan LIU ; Liyu LIU ; Ling ZHANG ; Wenxuan LIU ; Ye QUAN ; Bohan MA ; Sakurai REIKO ; Virender Kumar REHAN
Journal of Beijing University of Traditional Chinese Medicine 2025;48(8):1167-1175
Objective To investigate and compare the effects of electroacupuncture and dexamethasone on lung function and histomorphology and behavior in the offspring of perinatal nicotine exposure(PNE)rats.Methods Twenty-four pregnant Sprague-Dawley rats were randomly divided into control,model,electroacupuncture shallow needling,and dexamethasone groups using the random number table method(n=6 rats per group).A pulmonary dysplasia model in offspring rats with PNE was prepared by cervicodorsal subcutaneous injection with nicotine,and successful establishment of the model was confirmed by demonstrating statistically significant differences in growth parameters,lung function,and lung morphology compared to the control group.From the sixth day of maternal conception,the control group received cervicodorsal subcutaneous injection with 0.9%sodium chloride solution(1 mg/kg once per day),and the remaining groups were similarly injected with nicotine(1 mg/kg once per day).Concurrent with the nicotine injections,the electroacupuncture shallow needling group received electroacupuncture interventions at the"Zusanli"(ST 36)acupoint on both sides once a day for 20 min.In contrast,the offspring rats in the dexamethasone group received intraperitoneal dexamethasone injections from postnatal day 8(1.5 mg/kg once per day for 3 days),with a later dose of 0.75 mg/kg once per day for 4 days.Following successful model establishment,four offspring per rat were retained in each group using the random number table method.Until the 21st day after the birth of the offspring,using the random number table method,one offspring randomly selected from the four retained offspring per rat in each group was assigned to growth parameter assessment including body weight,lung weight,and lung index;simultaneously,one offspring was randomly selected for pulmonary function which was detected by a small animal pulmonary function machine,with the parameters of forced vital capacity(FVC),functional residual capacity(FRC),dynamic lung compliance(Cdyn),peak expiratory flow(PEF),peak inspiratory flow(PIF),and airway resistance(RL);concurrently,one offspring was randomly selected for hematoxylin and eosin staining to observe the histomorphology of the lung tissue,alveolar numbers,and mean alveolar septal thickness;additionally,one offspring was randomly selected for Morris water maze testing to evaluate the effects of the two intervention methods on learning and memory capabilities in offspring rats with PNE.Results Compared with the control group,the body weight,lung weight,lung index,FVC,PEF,FRC,Cdyn,alveolar number,platform crossing times,target quadrant time percentage,and target quadrant path percentage in the Morris water maze experiment were significantly decreased in the offspring rats of the model group(P<0.01).In contrast,PIF,RL,average thickness of alveolar septa,and latency of the Morris water maze experiment were significantly increased in the offspring rats of the model group(P<0.01).Compared with the model group,both electroacupuncture shallow needling group and dexamethasone group exhibited significant increases in body weight,lung weight,lung index,FVC,PEF,FRC,Cdyn,and alveolar numbers(P<0.05,P<0.01),along with significant decreases in PIF,RL values,and mean alveolar septal thickness(P<0.01).However,in the Morris water maze experiment,compared with the model group,the electroacupuncture shallow needling group demonstrated significantly more platform crossing times and a higher target quadrant time percentage and target quadrant path percentage(P<0.01),coupled with reduced latency period(P<0.01),whereas the dexamethasone group showed no significant differences compared to the model group.Compared to the electroacupuncture shallow needling group,the dexamethasone group showed significantly less platform crossing times and a lower target quadrant time percentage and target quadrant path percentage,coupled with increased latency period(P<0.01).Conclusion Both electroacupuncture and dexamethasone demonstrated protective effects on lung development in PNE offspring rats by ameliorating lung histomorphology and pulmonary function.However,offspring receiving late,small-dose,short-course dexamethasone exhibited inferior learning and memory capabilities,whereas the electroacupuncture group showed significantly superior cognitive performance compared to the dexamethasone group.
4.Comparison of electroacupuncture and dexamethasone on lung development impairment in offspring rats induced by prenatal nicotine exposure
Tingting GUO ; Bo JI ; Yang FANG ; Yitian LIU ; Hewen LI ; Xiaoxuan LIU ; Liyu LIU ; Ling ZHANG ; Wenxuan LIU ; Ye QUAN ; Bohan MA ; Sakurai REIKO ; Virender Kumar REHAN
Journal of Beijing University of Traditional Chinese Medicine 2025;48(8):1167-1175
Objective To investigate and compare the effects of electroacupuncture and dexamethasone on lung function and histomorphology and behavior in the offspring of perinatal nicotine exposure(PNE)rats.Methods Twenty-four pregnant Sprague-Dawley rats were randomly divided into control,model,electroacupuncture shallow needling,and dexamethasone groups using the random number table method(n=6 rats per group).A pulmonary dysplasia model in offspring rats with PNE was prepared by cervicodorsal subcutaneous injection with nicotine,and successful establishment of the model was confirmed by demonstrating statistically significant differences in growth parameters,lung function,and lung morphology compared to the control group.From the sixth day of maternal conception,the control group received cervicodorsal subcutaneous injection with 0.9%sodium chloride solution(1 mg/kg once per day),and the remaining groups were similarly injected with nicotine(1 mg/kg once per day).Concurrent with the nicotine injections,the electroacupuncture shallow needling group received electroacupuncture interventions at the"Zusanli"(ST 36)acupoint on both sides once a day for 20 min.In contrast,the offspring rats in the dexamethasone group received intraperitoneal dexamethasone injections from postnatal day 8(1.5 mg/kg once per day for 3 days),with a later dose of 0.75 mg/kg once per day for 4 days.Following successful model establishment,four offspring per rat were retained in each group using the random number table method.Until the 21st day after the birth of the offspring,using the random number table method,one offspring randomly selected from the four retained offspring per rat in each group was assigned to growth parameter assessment including body weight,lung weight,and lung index;simultaneously,one offspring was randomly selected for pulmonary function which was detected by a small animal pulmonary function machine,with the parameters of forced vital capacity(FVC),functional residual capacity(FRC),dynamic lung compliance(Cdyn),peak expiratory flow(PEF),peak inspiratory flow(PIF),and airway resistance(RL);concurrently,one offspring was randomly selected for hematoxylin and eosin staining to observe the histomorphology of the lung tissue,alveolar numbers,and mean alveolar septal thickness;additionally,one offspring was randomly selected for Morris water maze testing to evaluate the effects of the two intervention methods on learning and memory capabilities in offspring rats with PNE.Results Compared with the control group,the body weight,lung weight,lung index,FVC,PEF,FRC,Cdyn,alveolar number,platform crossing times,target quadrant time percentage,and target quadrant path percentage in the Morris water maze experiment were significantly decreased in the offspring rats of the model group(P<0.01).In contrast,PIF,RL,average thickness of alveolar septa,and latency of the Morris water maze experiment were significantly increased in the offspring rats of the model group(P<0.01).Compared with the model group,both electroacupuncture shallow needling group and dexamethasone group exhibited significant increases in body weight,lung weight,lung index,FVC,PEF,FRC,Cdyn,and alveolar numbers(P<0.05,P<0.01),along with significant decreases in PIF,RL values,and mean alveolar septal thickness(P<0.01).However,in the Morris water maze experiment,compared with the model group,the electroacupuncture shallow needling group demonstrated significantly more platform crossing times and a higher target quadrant time percentage and target quadrant path percentage(P<0.01),coupled with reduced latency period(P<0.01),whereas the dexamethasone group showed no significant differences compared to the model group.Compared to the electroacupuncture shallow needling group,the dexamethasone group showed significantly less platform crossing times and a lower target quadrant time percentage and target quadrant path percentage,coupled with increased latency period(P<0.01).Conclusion Both electroacupuncture and dexamethasone demonstrated protective effects on lung development in PNE offspring rats by ameliorating lung histomorphology and pulmonary function.However,offspring receiving late,small-dose,short-course dexamethasone exhibited inferior learning and memory capabilities,whereas the electroacupuncture group showed significantly superior cognitive performance compared to the dexamethasone group.
5.The Scientific Connotation of"Heart and Brain Co-Dominating the Mind"and Its Application in the Treatment of Insomnia by Acupuncture
Yuting ZHANG ; Shan QIN ; Xiaoqiu WANG ; Liyu LIN ; Mufeng LI ; Wenzhong WU ; Chengyong LIU
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):181-188
The paper discusses the scientific connotation of"heart and brain co-dominating the mind",including the connotation of"heart and brain co-dominating the mind"in traditional Chinese medicine and the physiological and pathological relationship be-tween heart and brain in modern medicine.Based on the understanding of"heart and brain co-dominating the mind",this paper be-lieves that"restlessness of mind"is the core pathogenesis of insomnia,and regulating mind is the key to the acupuncture and moxibus-tion treatment of insomnia.The autonomic nervous system(ANS)is a bridge connecting the heart and brain,and is closely related to the central anatomy,neurotransmitters and physiological rhythms of sleep.The imbalance of ANS is highly consistent with the patho-genesis of"heart and brain restlessness"in traditional Chinese medicine.The Governor Vessel runs through the brain and the heart,and the"Governor Vessel regulating spirit acupuncture"can communicate the heart and brain,regulate ANS function,and thus im-prove sleep quality.
6.Prediction of invasiveness in follicular variant of papillary thyroid carcinoma using nomograms based on ultrasonic features
YuXin ZHENG ; Yajiao ZHANG ; Liyu CHEN ; Kefeng LU ; Jiangyan LOU ; Junping LIU ; Dong XU
Chinese Journal of Ultrasonography 2024;33(9):800-806
Objective:To explore the relationship between ultrasound characteristics and invasiveness in the follicular variant of papillary thyroid carcinoma (FVPTC), and to integrate multiple ultrasound parameters for visual assessment of predictive outcomes by using Nomogram.Methods:A total of 312 FVPTC patients who were pathologically confirmed through surgery in Zhejiang Cancer Hospital and Zhejiang Provincial People′s Hospital from January 2013 to December 2023 were retrospectively collected.Based on defined criteria, FVPTC patients were categorized into high-invasion and low-invasion groups. The dataset was divided into a training set and a validation set in a ratio of 7 to 3. Clinical information and ultrasound feature parameters were collected. Univariate and multivariate Logistic regression analyses were performed on the training set. A predictive model for FVPTC invasiveness was constructed based on ultrasound features. The model′s discriminative ability and calibration were evaluated in the validation set, and a nomogram was generated.Results:The training set included a total of 218 patients with FVPTC, among which 131 were classified as high invasive.The validation set consisted of 94 patients, with 53 cases of high invasive FVPTC patients. Multivariate logistic regression analysis on the training set revealed that tumor multifocality ( OR=6.505, P=0.016), hypoechoic ( OR=3.235, P=0.103), shape ( OR=0.521, P=0.049), and microcalcifications ( OR=2.479, P=0.004) were independent influencing factors for predicting invasiveness in FVPTC. In the training set, the area under the curve (AUC) of the ultrasound predictive model was 0.704 (95% CI=0.634-0.771), and in the validation set, the AUC was 0.650 (95% CI=0.531-0.770), indicated good discriminative ability.The calibration curve showed good alignment with the ideal curve, demonstrating favorable calibration performance. Conclusions:Ultrasound features provide valuable information for assessing the invasiveness of FVPTC, and the model constructed by combining ultrasound features demonstrates good predictive efficacy for the invasiveness of FVPTC.
7.The safety and feasibility of peripheral vascular intervention via the ipsilateral transulnar access due to failure of transradial artery puncture
Weilin TIAN ; Xiaoxi MENG ; Huaqiang LIAO ; Hongchao LIU ; Yafeng GU ; Liyu HUANG ; Weihua DONG ; Hailin JIANG
Journal of Interventional Radiology 2024;33(7):723-727
Objective To investigate the safety and feasibility of peripheral vascular intervention via the ipsilateral transulnar access(TUA)due to failure of transradial access(TRA)puncture.Methods The clinical data of 2546 peripheral vascular interventions via TRA,which were performed at authors'hospital between January 2019 and December 2021,were retrospectively analyzed.Among the 2546 interventions,TRA puncture failed in 37 procedures,and in 27 of these patients the ipsilateral TUA puncture had to be adopted.The puncture success rate,surgical success rate and puncture approach-related complications of TUA of the 27 patients receiving ipsilateral TUA puncture were analyzed.Results The success rate of ipsilateral TUA puncture after TRA puncture failed was 96.3%(26/27),and in one patient transfemoral access(TFA)puncture had to be substituted because of the ulnar artery spasm.The total success rate of interventional procedures was 96.3%(26/27).No serious complications occurred,and the incidence of minor complications was 19.2%(5/26).Conclusion Preliminary results indicate that for the experienced TRA operators,using ipsilateral TUA puncture due to failure of TRA puncture is a safe and feasible strategy choice.
8.Non-targeted Metabolomics Analysis of Fuling Yunhua Granules in Treatment of Type 2 Diabetes Mellitus Rats
Mengyao TIAN ; Keke LUO ; Mengxiao WANG ; Tianbao HU ; Hongmei LI ; Zongyuan HE ; Lixin YANG ; Liyu HAO ; Nan SI ; Yuyang LIU ; Baolin BIAN ; Hongjie WANG ; Yanyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):195-204
ObjectiveBased on non-targeted metabolomics, to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus(T2DM) rats by Fuling Yunhua granules, and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats, half male and half female, were randomly divided into the control group, model group, and high, medium, low dose groups of Fuling Yunhua granules(20.70, 10.35, 5.18 g·kg-1 in raw drug amount) and the positive drug group(pioglitazone hydrochloride tablets, 8.1 mg·kg-1). Except for the control group, other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ) to establish a T2DM rat model. After successful modeling, the treatment groups were administered the corresponding drugs by gavage, and the control group and model group were treated with an equal volume of saline by gavage, once/d, for 28 d. Fasting blood glucose(FBG) and glycosylated hemoglobin A1c(GHbA1c) levels were measured in all groups of rats during the administration period, and hematoxylin-eosin(HE) staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-LTQ-Orbitrap MS), and the data were processed using principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Differential metabolites were identified by the Human Metabolome Database(HMDB) and the Kyoto Encyclopedia of Genes and Genomes(KEGG), and screened for differential metabolites with variable importance in the projection(VIP) value>1, P<0.05, and fold change(FC)<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0, then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic(ROC) curves. ResultCompared with the control group, the FBG level of rats in the model group increased significantly(P<0.01), the GHbA1c content tended to increase, but the difference was not statistically significant, and the pancreatic tissue of rats was obviously damaged, the number of pancreatic islets decreased, and the pancreatic β-cells were obviously reduced, atrophied and enlarged. Compared with the model group, the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration(P<0.05, P<0.01), the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced(P<0.05), and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid, D-amino acid metabolism, steroid hormone biosynthesis, and glycerophospholipid metabolism in rats. Compared with the model group, the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted, and the pathway enrichment analysis found that D-amino acid metabolism, retinol metabolism, glycine, serine and threonine metabolism, tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol, D-piperidinic acid, D-serine, and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.
9.Clinical and Animal Experimental Research on Kidney Deficiency Syndrome: A Review
Guimin LIU ; Jianhui SUN ; Jianliang LI ; Zeyue YU ; Liyu HAO ; Hanhui HUANG ; Zongyuan LI ; Aoao WANG ; Yang YANG ; Hongmei LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):269-280
Kidney deficiency syndrome is a common clinical syndrome in traditional Chinese medicine (TCM). With the progress of science and technology, clinical and animal experiments on kidney deficiency syndrome have made remarkable progress. Research on kidney deficiency and the nature of "kidney" involves a large number of physiological and pathological bases, which are closely related to physiological and pathological links in the human body, among which the neuroendocrine-immune network shares the closest relationship. However, there are still many challenges in modern research on kidney deficiency syndrome, such as expert consensus on clinical diagnostic criteria and evaluation indexes and optimization of animal experimental models. In the past decade, a large number of clinical and animal experiments have been reported in the literature on kidney deficiency syndrome, among which the literature focusing on the combination of disease and syndrome is predominant, and most of them focus on kidney Yang deficiency and kidney Yin deficiency, involving the exploration of many pathological mechanisms. Research on the mechanisms related to kidney deficiency syndrome encompasses multiple signaling pathways and various biochemical indicators, including the phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-erythroid 2-relatedfactor-2(PI3K/Akt/Nrf2) signaling pathway, the Toll-like receptor 4/myeloid differentiation factor88/nuclear factor-κB(TLR4/MyD88/NF-κB) signaling pathway, the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signaling pathway, Osteoprotectin/nuclear factor-κB receptor activator ligand/receptor activator of nuclear factor-κB (OPG/RANKL/RANK) signaling pathway. The biochemical indicators cover the cyclic adenosine monophosphate/cyclic guanosine monophosphate (cAMP/cGMP) ratio, Na+-K+-ATPase activity, Ca2+-Mg2+-ATPase activity, adrenocorticotropic hormone (ACTH), polycorticosterone (CORT), 17-OHCS, and other sex hormone indicators, providing crucial reference values for diagnosing kidney Yang deficiency or kidney Yin deficiency. The literature related to kidney deficiency syndrome over the past decade was collated and excavated, with a view to providing a reference for research on kidney deficiency syndrome.
10.Clinical Manifestations,Molecular Genetics and Gonadal Pathology of 416 Patients with Disorders of Sex Development:A Single-Center Cohort Study
Wanjun LIN ; Cuili LIANG ; Wen FU ; Liyu ZHANG ; Wei JIA ; Jinhua HU ; Wen ZHANG ; Yunting LIN ; Huilin NIU ; Liping FAN ; Zhikun LU ; Duan LI ; Zongcai LIU ; Huiying SHENG ; Xi YIN ; Xiaodan CHEN ; Guochang LIU ; Jing CHENG ; Li LIU
JOURNAL OF RARE DISEASES 2024;3(3):310-317
Objective To investigate the clinical manifestations,molecular genetics and gonadal pathol-ogy characteristics of patients with disorders of sex development(DSD),and to summarize the clinical experi-ence of identifying rare diseases from common symptoms.Methods The clinical data of 416 patients with DSD diagnosed and treated in the multidisciplinary center of DSD of Guangzhou Women and Children's Medical Cen-ter from May 2018 to August 2023 were retrospectively analyzed,summarized and discussed.Results Accord-ing to chromosome karyotype,416 cases of DSD were classified into three types:92 cases(22.1%)of abnormal sex chromosome karyotype,285 cases(68.5%)of 46,XY karyotype and 39 cases(9.4%)of 46,XX karyotype.Among the 92 patients with abnormal sex chromosome karyotype,59 cases were raised as males,18 cases(30.5%)complained of short penis with hypospadias and cryptorchidism.The most common karyotype was 45,X/46,XY(58 cases,63.0%).Among the 285 patients with 46,XY karyotype,238 cases were raised as males,and 63 cases(26.5%)complained of short penis and hypospadias;47 cases were raised as females,and 13 ca-ses(27.7%)complained of inguinal mass.A total of 216 patients with 46,XY karyotype were subjected to whole exome gene detection,and 155 cases(71.8%)were found to have molecular pathogenesis with the clinical phe-notype.Among the 39 patients with 46,XX karyotype,19 cases were raised as males,and 8 cases(42.1%)com-plained of short penis and hypospadias.In the 18 cases of gonad biopsy,17 cases showed testicular tissue in go-nads.Whole exome sequencing was performed in 14 cases.NR5A1 gene heterozygous mutation,SRY gene muta-tion and SOX3 gene mutation were found in 2 cases,respectively(14.3%).Twenty cases were raised as females,and 14 cases(70.0%)complained of clitoral hypertrophy.Gonad biopsy was performed in 8 cases,with 7 cases of ovotestis(87.5%)and 1 case of NR5A1 gene heterozygous mutation(14.3%).Conclusions The etiologies of DSD are complex and diverse,and the clinical manifestations are various,which can be manifested as hypospa-dias,micropenis,cryptorchidism and other common symptoms of the urinary system.Different etiologies have dif-ferent treatment options.Therefore,chromosome karyotype,molecular genetic testing and gonadal pathology can be used to clarify the cause of disease,especially for rare diseases,improve the detection rate,reduce the rate of missed diagnosis,and ensure reasonable treatment,especially sex selection.

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