Objective·To establish a transgenic zebrafish line Tg(amh:mCherry)for studying gonadal development and related diseases,and to trace and analyze the developmental process of zebrafish gonads.Methods·A recombinant transgenic expression vector pTol2-amh-mCherry was constructed by using the upstream promoter sequence of the zebrafish anti-Müllerian hormone(amh)gene coding region and the gene coding sequence of the red fluorescent protein,and was validated by using Sanger sequencing.The recombinant transgenic expression vector and mRNA of Tol2 transposase were co-microinjected into zebrafish embryos,and fluorescent zebrafish were selected by using fluorescence microscope to screen for a stable inherited transgenic zebrafish line Tg(amh:mCherry).In situ hybridization was used to detect whether the location of the fluorescence signals was consistent with the location of endogenous expression of amh.The red fluorescence of transgenic zebrafish from generation F3 was observed by fluorescence microscopy,and the process of gonadal development was tracked and analyzed.Results·The recombinant transgenic expression vector pTol2-amh-mCherry was successfully constructed,and a stable inherited F3 generation transgenic zebrafish line Tg(amh:mCherry)was successfully established.The results of in situ hybridization showed that the location of the red fluorescence signals in the transgenic zebrafish Tg(amh:mCherry)was consistent with the location of endogenous expression of amh.Through observation and analysis of the fluorescence of transgenic zebrafish from F3 generation,five developmental patterns of zebrafish gonads were found.Conclusion·The successful construction of Tg(amh:mCherry)transgenic zebrafish line facilitates the tracer analysis of zebrafish gonadal development,which provides a better experimental model for the study of gonad-related diseases.

Objective·To establish a transgenic zebrafish line Tg(amh:mCherry)for studying gonadal development and related diseases,and to trace and analyze the developmental process of zebrafish gonads.Methods·A recombinant transgenic expression vector pTol2-amh-mCherry was constructed by using the upstream promoter sequence of the zebrafish anti-Müllerian hormone(amh)gene coding region and the gene coding sequence of the red fluorescent protein,and was validated by using Sanger sequencing.The recombinant transgenic expression vector and mRNA of Tol2 transposase were co-microinjected into zebrafish embryos,and fluorescent zebrafish were selected by using fluorescence microscope to screen for a stable inherited transgenic zebrafish line Tg(amh:mCherry).In situ hybridization was used to detect whether the location of the fluorescence signals was consistent with the location of endogenous expression of amh.The red fluorescence of transgenic zebrafish from generation F3 was observed by fluorescence microscopy,and the process of gonadal development was tracked and analyzed.Results·The recombinant transgenic expression vector pTol2-amh-mCherry was successfully constructed,and a stable inherited F3 generation transgenic zebrafish line Tg(amh:mCherry)was successfully established.The results of in situ hybridization showed that the location of the red fluorescence signals in the transgenic zebrafish Tg(amh:mCherry)was consistent with the location of endogenous expression of amh.Through observation and analysis of the fluorescence of transgenic zebrafish from F3 generation,five developmental patterns of zebrafish gonads were found.Conclusion·The successful construction of Tg(amh:mCherry)transgenic zebrafish line facilitates the tracer analysis of zebrafish gonadal development,which provides a better experimental model for the study of gonad-related diseases.

Methylmalonic acidemia (MMA) is a series of rare inherited organic acid metabolic disorders with variable and nonspecific clinical manifestations, in particular neurological symptoms such as vomiting, lethargy, etc. Even with timely treatment, patients may still have various degrees of neurological complications and can even die. The prognosis is mainly related to the type of genetic variants, level of metabolites, newborn screening, onset of disease and early initiation of treatment. This article has reviewed the prognosis of patients with various types of MMA and factors that may affect it.
Infant, Newborn ; Humans ; Amino Acid Metabolism, Inborn Errors/complications* ; Prognosis ; Mutation ; Neonatal Screening ; Propionic Acidemia

Objective:To construct a combined diet and exercise program for puerperal women with Yin-deficiency constitution based on traditional Chinese medicine (TCM) constitution identification, and evaluate the effect of this program.Methods:This study was guided by the complex intervention development framework of the UK Research Council, and based on the TCM constitution identification, and determined the combined diet and exercise program for puerperal women with Yin-deficiency constitution based on TCM constitution identification through literature review, case analysis, group discussion, and Delphi method. From October to December 2022, convenience sampling was used to select 76 delivery women with Yin-deficiency constitution who delivered at Shenzhen Hospital of Southern Medical University as the research subject. According to the delivery time, the delivery women were divided into a control group ( n=37) and an experimental group ( n=39). The control group adopted routine postpartum nursing. On this basis, the experimental group implemented the combined diet and exercise program for puerperal women with Yin-deficiency constitution based on TCM constitution identification. We compared the Yin-deficiency constitution score, duration of lochia, and incidence of puerperium complications between two groups of delivery women before and after intervention. Results:The final constructed program included 6 primary indicators, 20 secondary indicators, and 50 tertiary indicators. In the two rounds of consultation, the positivity coefficients of experts were all 100%, the authority coefficients of experts were 0.827 and 0.843, and the Kendall harmony coefficients were 0.419 and 0.502, respectively with a statistical difference ( P<0.01). After intervention, the score of Yin-deficiency constitution, duration of lochia, and incidence of puerperium complications in the experimental group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The combined diet and exercise program for puerperal women with Yin-deficiency constitution based on TCM constitution identification is scientific, comprehensive, and feasible, which can provide reference for future development of TCM constitution management during the puerperium.

Objective:To explore the effect of Rain Classroom combined with situational teaching in clinical clerkship of internal medicine.Methods:A total of 72 undergraduate students with clinical majors of Batch 2015 were selected and randomly divided into a control group and an experimental group using computer scrolling pause method, with 36 students in each group. The control group adopted traditional teaching methods, and the research group adopted Rain Classroom combined with situational teaching. The teaching effect evaluation was carried out through a 10-point questionnaire based on the internal medicine theory examination, clinical skill assessment and learning interest, knowledge mastery and application, and clinical thinking and practical ability. SPSS 20.0 was used to conduct t-test. Results:The average scores of the students in the experimental group including the theoretical test (87.94±5.68) and the skill test scores (88.08±5.73) were significantly higher than those in the control group (83.00±5.89) and (83.86±4.80)( P<0.01). The results of the questionnaire showed that students' interest in learning improvement, self-learning ability, new knowledge acquisition, application of knowledge, clinical thinking, clinical practice, problem solving, self-expression, teamwork, doctor-patient communication ability in the study group were better than those in the control group ( P < 0.01). Conclusion:The Rain Classroom combined with situational teaching method is helpful to improve students' knowledge mastery and application, improve clinical thinking and practical ability, improve learning interest and clinical clerkship teaching effect.

Objective:To apply the best evidence of perioperative enhanced recovery nursing for lung cancer patients to clinical practice, and to evaluate the effect of practice.Methods:Following the 5 steps of the Stetler model of research utilization, an evidence-based protocol was formed through literature search, quality assessment, evidence synthesis and revision by expert group meetings. From January to July 2021, convenience sampling was used to select 160 lung cancer patients admitted to the Thoracic Surgery of Shanghai Chest Hospital as the research object. Patients admitted from January to April 2021 were set as the control group ( n=80) for baseline review. Patients admitted from May to July 2021 were set as the intervention group ( n=80) , and an evidence-based protocol was applied. Results:Finally, one guideline and 7 consensus articles were included. The evidence-based protocol covered 10 modules of preoperative nursing assessment, nutritional management, dietary preparation, health education, postoperative dietary management, functional exercise, pipeline management, pain management, venous thrombosis management and symptom management, with a total of 22 recommendations. There were significant differences in the time of first ambulation, the completion of ambulation on the day of operation, the pain score of first ambulation, the time for the first eating, and the time for the first exhausting between the two groups ( P<0.05) . There were no complications and venous thrombosis in the two groups during hospitalization. Conclusions:The development of the evidence-based practice project is beneficial to accelerate the postoperative recovery of lung cancer patients, and is conducive to standardizing the nursing path, thereby promoting the improvement of nursing quality.

Objective:To explore the clinical characteristics, genotypes and long-term outcomes of individuals with 3-methylglutaconic aciduria.Methods:The clinical features, biochemical data, genetic test results and treatment outcomes of six children with 3-methylglutaconic aciduria admitted to the Department of Endocrinology, Genetics and Metabolism, Xinhua Hospital from February 2017 to February 2019 were retrospectively analyzed and the Gesell developmental diagnosis schedule was performed to evaluate the development of four patients.Results:Among 6 children with 3-methylglutaconic aciduria 2 were males and 4 were females.Four cases had 3-methylglutaconic aciduria type Ⅰ and 2 cases had 3-methylglutaconic aciduria with deafness,encephalopathy, and Leigh-like syndrome. Five of 6 patients were detected by newborn screening among whom 4 remained asymptomatic, and only one had a postmortem diagnosis. Among them, 4 patients remained asymptomatic, while two presented with clinical symptoms such as jaundice and dyspnea and the age of disease onset was 1 and 2 days respectively. The concentration of 3-methylglutaconic acid in urine of all affected individuals was between 22.38 and 77.09 mmol/molCr, which was above the normal value. Genetic tests were performed for all patients. Eleven variants were identified in 2 genes, of which 10 variants were novel and only c.442C>T p.(R148X) has been previously reported; Seven variants (c.656-2delA, EX5-EX6 Del, c.942+3A>G, c.373C>T p.(R125W), c.895-3C>G, c.667C>T p.(R223X) and c.894+5G>A) were in AUH gene. The others (c.548G>A p.(R138Q), c.442C>T p.(R148X), c.1339C>T p.(R447X) and c.973dupA p.(M325Nfs*5) were in SERAC1 gene. After being treated with leucine diet restriction and L-carnitine, 4 patients with AUH gene variation who were from asymptomatic phase developed normally, whereas those 2 patients with SERAC1 gene variation had a poor prognosis. During the follow-up, 2 patients exhibited varying degrees of psychomotor retardation, the rest had normal course of development.Conclusions:There are significant clinical heterogeneities among individuals with 3-methylglutaconic aciduria. The most common pathogenic variants are splicing variations, followed by nonsense, missense and frameshift mutations. Leucine-free diet and oral L-carnitine therapy are effective for some patients. Newborn screening is essential for early diagnosis and improvement of prognosis.

Objective To explore the efficacy of moderate dose bromocriptine by one-off oral administration in treatment of patients with non-acute prolactin (PRL) type pituitary adenoma.Methods Forty-three patients with definite diagnosis of non-acute prolactin type pituitary adenoma,admitted to our hospital from January 2016 to December 2017,were chosen in our study.All patients were administrated with 5 mg bromocriptine at 18 pm;the serum PRL levels were examined at 2,3 and 12 h after bromocriptine administration.The curative effects of these patients were judged by decline rate of PRL,and the curative effects of patients with different genders,different initial PRL levels and different ages were compared.Results In these 43 patients,bromocriptine showed excellent effect in 20 patients,obvious effect in 18 and invalid effect in 5.There were statistically significant differences in the efficacy of bromocriptine between different genders (P＜0.05).There were statistically significant differences in efficacy of patients with different genders after taking bromocriptine (P＜0.05).The female patients had obviously higher rate of obvious effect than the male patients.Patients with different initial PRL values had statistically significant differences in efficacy after taking bromocriptine (P＜0.05).Patients with PRL initial value of 40-200 ng/mL had significantly higher rate of obvious effect than patients with PRL initial value of ＞200 ng/mL.There was no significant difference in efficacy between patients ＜ 45 years old and patients＞45 years old (P＞0.05).Conclusion The treatment ofhyperprolactin caused by prolactin type pituitary adenoma with moderate dose bromocriptine by one-off oral administration is effective;female patients have obviously better effect than male patients;and for patients with initial prolactin＞200 ng/mL,the effect is better.

Objective To explore prevention and treatment of complications following resection of pineal lesions via the modified Poppen approach.Methods From January 2008 to December 2017,62 patients received resection of pineal region lesions via the modified Poppen approach at Department of Neurosurgery,Anhui Provincial Hospital.Their clinical data and outcomes were reviewed.The incidences of postoperative complications were compared between the patients with differences in lesion diameter,tentorial angle and preoperative hydrocephalus.Results Of this cohort,total tumor resection was performed in 48,subtotal resection in 10 and partial resection in 4.Follow-ups from 3 to 12 months revealed 2 cases of hemianopia which were recovered 3 months later,7 cases of occipital lobe contusion,2 cases of monoparesis and 2 cases of intracranial infection.Of the 7 cases of occipital lobe contusion,3 had intracranial hematoma which had been removed by secondary surgery and 4 had ataxia.The incidences of postoperative hemianopia and monoparesis for the patients with a lesion ＞4.5 cm in diameter were significantly higher than for those with a lesion ≤4.5 cm in diameter (P＜0.05).The incidence of postoperative hemianopia for the patients with a tentorial angle ＞60° was significantly higher than for those with a tentorial angle ≤ 60° (P＜0.05).Conclusions Size and tentorial angle of the lesion may be closely associated with the complications following resection of pineal lesions via the modified Poppen approach.The key to reducing the postoperative complications lies in comprehensive preoperative assessments,familiarity with clinical anatomy of the Poppen approach,sophisticated surgical skills and an individualized surgical protocol.

Objective To conclude the application experience of intraoperative ultrasound added with neuronavigation in resection of intracranial gliomas.Methods A retrospective analysis was performed on the clinical data of 60 patients with intracranial gliomas,performed resection in our hospital from January 2010 to June 2013 by intraoperative ultrasound added with neuronavigation; 32 of them had high-grade gliomas and 28 low-grade ones.Ultrasonoscopy of different grade gliomas was studied.Results The accuracy of localization was 100％ in 60 patients with intracranial gliomas.Ultrasonic image of high-grade gliomas enjoyed obvious hyperechogenicity and the boundaries were clear usually; ultrasonic image of low-grade ones were slightly hyperechogenic or isoechogenic and the boundaries were indistinct usually.Inhomogeneous hyperechogenic of the ultrasonoscopy of intracranial gliomas before resection and hypoechogenic of residual cavity after resection were noted,respectively.Total removal was achieved in 50 patients,subtotal resection in 6 and partial resection in 4.Brain swelling happened postoperatively in 1 and was remitted by decompressive craniectomy.Hemiplegia happened in 2,and no death occurred postoperatively.Conclusions Intraoperative ultrasound is important to the correct of brain shift,and has clear and important value to the choice of operation approach and incising cortex.Ultrasonic image of high-grade gliomas and calcified lesions enjoys the best view.Intraoperative ultrasound added with neuronavigation can raise resection rate of intracranial gliomas.