1.Acute isolation of caudate nucleus neurons with patch-clamp technique in neonate rats
Congde LI ; Yongli LU ; Hongwei YANG ; Min ZHOU
Chinese Journal of Tissue Engineering Research 2007;0(37):-
BACKGROUND: Single scattered neurons are needed in most studies on ion channels. However, artificial primary cultured neurons, for the influence from the environment, have a lot of changes in their characteristics. Comparatively, acutely isolated neurons can keep intact physiological characteristics. OBJECTIVE: To establish an ideal method for acute isolation of caudate nucleus neurons and to study their ion channels and signal transduction mechanism with patch clamp technique for the benefit of deeply understanding functions of caudate nucleus. DESIGN, TIME AND SETTING: An observational experiment on cells was performed at the Central Laboratory, Medicine Science College of China Three Gorges University from February to December in 2008. MATERIALS: Twelve 7- to 10-day-old neonatal Wistar rats, of either gender, were selected for this study. METHODS: Scattered single caudate nucleus neuron was obtained from 7- to 10-day-old neonatal Wistar rats with a combination of enzymatic and mechanical separation method. Voltage-dependent calcium currents were determined with whole-cell patch clamp technique MAIN OUTCOME MEASURES: Observation of morphology and electrophysiological characteristics of acutely isolated caudate nucleus neurons with whole-cell patch clamp. RESULTS: The caudate nucleus neuron acutely isolated with protease digestion and mechanical isolation method was found to have regular configuration, intact cell membrane, long neuraxons, excellent morphology and physiological nature. Their L-calcium ion channel current observation showed electrical parameters within normal physiological range, and activity of voltage-dependent calcium ion channel was well preserved. CONCLUSION: As neurons isolated in this way have normal morphology and long neuraxons, and activities of primary iron channels are well preserved, we conclude that an acute isolation method of caudate nucleus neurons from neonate rats with patch clamp technique is successfully established.
2.Correction of virilization of external genitalia in children with congenital adrenal hyperplasia
Liguang XIA ; Weiwu PAN ; Weizhong ZHOU ; Weite QIAN ; Xiaokun LIN ; Zhongrong LI ; Congde CHEN
Chinese Journal of Plastic Surgery 2022;38(8):873-880
Objective:To summarize and share the experience of one-stage feminization surgery in children with congenital adrenal hyperplasia (CAH).Methods:The clinical data of CAH female children in the Department of Pediatric Urology, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from January 2018 to December 2020 were retrospectively analyzed. According to the Prader classification standard, the external genital virilism of children was graded. The cases of Prader grades Ⅱ to Ⅳ were underwent clitoroplasty and labiaplasty, while the cases of Prader Ⅱ and Ⅳ grades were required further vaginoplasty.Partial urogenital sinus mobilization(PUM) or total urogenital sinus mobilization(TUM )was performed according to the length of the urethrovaginal junction to the vulvar opening during the procedure. The postoperative follow-up was included the recovery of the external genitalia wound, the child’s ability to control urination, and the family members’ satisfaction with the child’s external genitalia appearance. Six months after the operation, the satisfaction with the appearance of the external genitalia was evaluated in the outpatient clinic, including four aspects: clitoris, labia majora, labia minora, and vaginal appearance.Results:A total of 18 female children were enrolled, ranging in age from 2 to 9 years old, with an average age of 4.5 years. There were 5 cases of Prader grade Ⅱ, 9 cases of grade Ⅲ, and 4 cases of grade Ⅳ. Ten cases (9 cases of Prader grade Ⅲ and 1 case of grade Ⅳ) underwent PUM, and 3 cases of Prader grade Ⅳ underwent TUM. All patients were followed up for 6-12 months after the operation, and all the wounds of the external genitalia of the children were healed in one stage. The satisfaction of family members to the appearance of external genitalia of children: 15 cases were satisfied, 3 cases were not satisfied; 3 patients were found to have overactive bladder two weeks after the operation, and no special treatment was given, the symptoms disappeared three months after the procedure, and the other children had no abnormal urination control ability. Six months after the process, the external genitalia was evaluated in the physician’s clinic: 17 cases were satisfied, and one was dissatisfied.Conclusions:The surgical method can be selected for better results according to the degree of external genitalia virilism and the common channel length in CAH female children.
3.Correction of virilization of external genitalia in children with congenital adrenal hyperplasia
Liguang XIA ; Weiwu PAN ; Weizhong ZHOU ; Weite QIAN ; Xiaokun LIN ; Zhongrong LI ; Congde CHEN
Chinese Journal of Plastic Surgery 2022;38(8):873-880
Objective:To summarize and share the experience of one-stage feminization surgery in children with congenital adrenal hyperplasia (CAH).Methods:The clinical data of CAH female children in the Department of Pediatric Urology, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from January 2018 to December 2020 were retrospectively analyzed. According to the Prader classification standard, the external genital virilism of children was graded. The cases of Prader grades Ⅱ to Ⅳ were underwent clitoroplasty and labiaplasty, while the cases of Prader Ⅱ and Ⅳ grades were required further vaginoplasty.Partial urogenital sinus mobilization(PUM) or total urogenital sinus mobilization(TUM )was performed according to the length of the urethrovaginal junction to the vulvar opening during the procedure. The postoperative follow-up was included the recovery of the external genitalia wound, the child’s ability to control urination, and the family members’ satisfaction with the child’s external genitalia appearance. Six months after the operation, the satisfaction with the appearance of the external genitalia was evaluated in the outpatient clinic, including four aspects: clitoris, labia majora, labia minora, and vaginal appearance.Results:A total of 18 female children were enrolled, ranging in age from 2 to 9 years old, with an average age of 4.5 years. There were 5 cases of Prader grade Ⅱ, 9 cases of grade Ⅲ, and 4 cases of grade Ⅳ. Ten cases (9 cases of Prader grade Ⅲ and 1 case of grade Ⅳ) underwent PUM, and 3 cases of Prader grade Ⅳ underwent TUM. All patients were followed up for 6-12 months after the operation, and all the wounds of the external genitalia of the children were healed in one stage. The satisfaction of family members to the appearance of external genitalia of children: 15 cases were satisfied, 3 cases were not satisfied; 3 patients were found to have overactive bladder two weeks after the operation, and no special treatment was given, the symptoms disappeared three months after the procedure, and the other children had no abnormal urination control ability. Six months after the process, the external genitalia was evaluated in the physician’s clinic: 17 cases were satisfied, and one was dissatisfied.Conclusions:The surgical method can be selected for better results according to the degree of external genitalia virilism and the common channel length in CAH female children.
4.Correction of virilization of external genitalia in children with congenital adrenal hyperplasia
Liguang XIA ; Weiwu PAN ; Weizhong ZHOU ; Weite QIAN ; Xiaokun LIN ; Zhongrong LI ; Congde CHEN
Chinese Journal of Plastic Surgery 2022;38(8):873-880
Objective:To summarize and share the experience of one-stage feminization surgery in children with congenital adrenal hyperplasia (CAH).Methods:The clinical data of CAH female children in the Department of Pediatric Urology, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from January 2018 to December 2020 were retrospectively analyzed. According to the Prader classification standard, the external genital virilism of children was graded. The cases of Prader grades Ⅱ to Ⅳ were underwent clitoroplasty and labiaplasty, while the cases of Prader Ⅱ and Ⅳ grades were required further vaginoplasty.Partial urogenital sinus mobilization(PUM) or total urogenital sinus mobilization(TUM )was performed according to the length of the urethrovaginal junction to the vulvar opening during the procedure. The postoperative follow-up was included the recovery of the external genitalia wound, the child’s ability to control urination, and the family members’ satisfaction with the child’s external genitalia appearance. Six months after the operation, the satisfaction with the appearance of the external genitalia was evaluated in the outpatient clinic, including four aspects: clitoris, labia majora, labia minora, and vaginal appearance.Results:A total of 18 female children were enrolled, ranging in age from 2 to 9 years old, with an average age of 4.5 years. There were 5 cases of Prader grade Ⅱ, 9 cases of grade Ⅲ, and 4 cases of grade Ⅳ. Ten cases (9 cases of Prader grade Ⅲ and 1 case of grade Ⅳ) underwent PUM, and 3 cases of Prader grade Ⅳ underwent TUM. All patients were followed up for 6-12 months after the operation, and all the wounds of the external genitalia of the children were healed in one stage. The satisfaction of family members to the appearance of external genitalia of children: 15 cases were satisfied, 3 cases were not satisfied; 3 patients were found to have overactive bladder two weeks after the operation, and no special treatment was given, the symptoms disappeared three months after the procedure, and the other children had no abnormal urination control ability. Six months after the process, the external genitalia was evaluated in the physician’s clinic: 17 cases were satisfied, and one was dissatisfied.Conclusions:The surgical method can be selected for better results according to the degree of external genitalia virilism and the common channel length in CAH female children.
5.Correction of virilization of external genitalia in children with congenital adrenal hyperplasia
Liguang XIA ; Weiwu PAN ; Weizhong ZHOU ; Weite QIAN ; Xiaokun LIN ; Zhongrong LI ; Congde CHEN
Chinese Journal of Plastic Surgery 2022;38(8):873-880
Objective:To summarize and share the experience of one-stage feminization surgery in children with congenital adrenal hyperplasia (CAH).Methods:The clinical data of CAH female children in the Department of Pediatric Urology, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University from January 2018 to December 2020 were retrospectively analyzed. According to the Prader classification standard, the external genital virilism of children was graded. The cases of Prader grades Ⅱ to Ⅳ were underwent clitoroplasty and labiaplasty, while the cases of Prader Ⅱ and Ⅳ grades were required further vaginoplasty.Partial urogenital sinus mobilization(PUM) or total urogenital sinus mobilization(TUM )was performed according to the length of the urethrovaginal junction to the vulvar opening during the procedure. The postoperative follow-up was included the recovery of the external genitalia wound, the child’s ability to control urination, and the family members’ satisfaction with the child’s external genitalia appearance. Six months after the operation, the satisfaction with the appearance of the external genitalia was evaluated in the outpatient clinic, including four aspects: clitoris, labia majora, labia minora, and vaginal appearance.Results:A total of 18 female children were enrolled, ranging in age from 2 to 9 years old, with an average age of 4.5 years. There were 5 cases of Prader grade Ⅱ, 9 cases of grade Ⅲ, and 4 cases of grade Ⅳ. Ten cases (9 cases of Prader grade Ⅲ and 1 case of grade Ⅳ) underwent PUM, and 3 cases of Prader grade Ⅳ underwent TUM. All patients were followed up for 6-12 months after the operation, and all the wounds of the external genitalia of the children were healed in one stage. The satisfaction of family members to the appearance of external genitalia of children: 15 cases were satisfied, 3 cases were not satisfied; 3 patients were found to have overactive bladder two weeks after the operation, and no special treatment was given, the symptoms disappeared three months after the procedure, and the other children had no abnormal urination control ability. Six months after the process, the external genitalia was evaluated in the physician’s clinic: 17 cases were satisfied, and one was dissatisfied.Conclusions:The surgical method can be selected for better results according to the degree of external genitalia virilism and the common channel length in CAH female children.