Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

Objective To analyze the postoperative effect of probiotics combined with Peitu Shengjin moxibustion and cupping on adenomatous colorectal polyps (ACP).Methods Patients with ACP diagnosed and treated in Zhejiang Provincial Hospital of Traditional Chinese Medicine from August 2021 to May 2022 were recruited as study subjects,and were divided into control group(probiotics) and test group (probiotics combined with Peitu Shengjin moxibustion and cupping).The main outcome measure of this study was the 1-year recurrence rate of ACP.The secondary outcome measures were the treatment efficiency and the traditional Chinese medicine (TCM) syndrome scores at 4 weeks and 1 year after ACP surgery.Results The study finally included 199 patients,with 100 in the control group and 99 in the test group.The 1-year postoperative treatment efficiency of the test group was higher than that of the control group (P=0.012);the recurrence rate was lower than that of the control group (P<0.05).There was no statistically significant difference in the comparison of polyp size and number between the two groups of patients with recurrence (P>0.05).Before treatment,there was no statistically significant difference between the two groups in the comparison of stool habit score,stool nature score,blood in stool score,stool frequency score,abdominal pain score and total score (P>0.05).The stool habit score,stool nature score,stool blood score,stool frequency score,abdominal pain score and total score of the patients in both groups were significantly lower than those of the preoperative period at 4 weeks and 1 year after the operation (P<0.05).Stool habit score,stool nature score,stool blood score,stool frequency score,abdominal pain score and total score were lower in the test group than in the control group at 4 weeks and 1 year postoperatively (P<0.05).Conclusion Probiotics combined with Peitu Shengjin moxibustion and cupping can effectively improve the TCM syndrome scores after ACP surgery and reduce the recurrence rate of ACP.

ACGS Best Practice Guidelines for Variant Classification in Rare Disease 2020, a supplementary practical guidelines, is based on the Standards and Guidelines for the Interpretation of Sequence Variations issued by the American Society for Medical Genetics and Genomics (ACMG) and the Association of Molecular Pathology (AMP) in 2015 by the British Medical Genetics Society under the Clinical Genomics Society (ACGS), and has integrated the detailed rules of standards developed by the ClinGen Sequence Variant Interpretation (SVI) Working Group by 2020. The further development of the ACMG/AMP guidelines is currently undertaken by the ClinGen SVI working group in the United States, which focuses on the classification of high penetrance and protein coding variants. ClinGen has established many expert panels on variants for specific diseases which required various evidence thresholds and is currently developing disease/gene specific guidelines. The British Medical Genetics Society has collected and integrated information on the guidelines for sequence variation classification and their extended rules, forming its own "2020 ACGS Best Practice Guidelines for Rare Disease Variation Classification" and is regularly updating it. The author has translated and summarized it for the reference of Chinese Medical Genetics Practitioners.
Humans ; Genetic Testing ; Genetic Variation ; Genome, Human ; Rare Diseases/genetics* ; China

Purpose: The unique chromosomal rearrangements of endometrial stromal sarcoma (ESS) make it possible to distinguish high-grade ESS (HGESS) and low-grade ESS (LGESS) from the molecular perspective. Analysis of ESS at the genomic and transcriptomic levels can help us achieve accurate diagnosis of ESS and provide potential therapy options for ESS patients. Materials and Methods: A total of 36 ESS patients who conducted DNA- and/or RNA-based next-generation sequencing were retrospectively enrolled in this study. The molecular characteristics of ESS at genomic and transcriptomic levels, including mutational spectrum, fusion profiles, gene expression and pathway enrichment analysis and features about immune microenvironment were comprehensively explored. Results: TP53 and DNMT3A mutations were the most frequent mutations. The classical fusions frequently found in HGESS (ZC3H7B-BCOR and NUTM2B-YWHAE) and LGESS (JAZF1-SUZ12) were detected in our cohort. CCND1 was significantly up-regulated in HGESS, while the expression of GPER1 and PGR encoding estrogen receptor (ER) and progesterone receptor (PR) did not differ significantly between HGESS and LGESS. Actionable mutations enriched in homologous recombination repair, cell cycle, and phosphoinositide 3-kinase/AKT/mammalian target of rapamycin pathways were detected in 60% of HGESS patients. Genes with up-regulated expression in HGESS were significantly enriched in five immune-related pathways. Most HGESS patients (85.7%) had positive predictors of immunotherapy efficacy. Moreover, immune microenvironment analysis showed that HGESS had relatively high immune infiltration. The degree of immune infiltration in HGESS patients with ZC3H7B-BCOR fusion was relatively higher than that of those with NUTM2B-YWHAE fusion. Conclusion This study investigated the molecular characteristics of ESS patients at the genomic and transcriptomic levels and revealed the potentially high sensitivity of targeted therapy and immunotherapy in a subset of HGESS with specific molecular features, providing a basis for guiding decision-making of treatment and the design of future clinical trials on precision therapy.

Objective:To explore the effect of health management based on constitution identification in perimenopausal women.Methods:From November 2020 to April 2021, 244 perimenopausal women from Doumen Street, Yuecheng District, Shaoxing were selected as the research subject by convenience sampling method, and divided into the control group (121 cases) and the observation group (123 cases) according to their respective communities. The control group adopted routine health management, while the observation group implemented health management based on constitution identification on the basis of routine health management. Three months after intervention, the perimenopausal symptom scores between the two groups were compared. One year after intervention, the number of individuals with biased constitution and quality of life scores between the two groups were compared.Results:Three months after intervention, the scores of hot flashes, sweating, insomnia, dizziness, fatigue, muscle and joint pain, headache, palpitations, and total score of perimenopausal symptom of the observation group were lower than those of the control group, with statistically significant differences ( P<0.05). Three months after intervention, there were no significant differences in scores of paraesthesia, mood swings, depressive suspicion, skin ant sensation, sexual difficulties, urinary tract infection between the observation group and the control group ( P>0.05). One year after intervention, the number of individuals with biased constitution of the observation group was less than that of the control group, and the difference was statistically significant ( P<0.05). One year after intervention, the scores of physical health, mental health, social relationship, and total score of quality of life of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05). One year after intervention, there was no statistically significant difference in the score of the surrounding environment between the observation group and the control group ( P>0.05) . Conclusions:Health management based on constitution identification can improve the perimenopausal symptoms of perimenopausal women, gradually change their biased constitution towards a calm constitution, and improve their quality of life.

Objective:To investigate cognitive function changes and their influential factors in patients with ischemic stroke and leukoaraiosis.Methods:A total of 500 patients with ischemic stroke who received treatment in Yiwu Central Hospital from January 2018 to October 2019 were included in this study. They were divided into simple ischemic stroke group ( n = 200) and ischemic stroke complicated by leukoaraiosis group (combination group, n = 300). The infarct location and the degree of leukoaraiosis in the combination group were analyzed. An additional 150 volunteers who concurrently underwent the Cognitive Function Test in the same hospital were selected as controls. Cognitive function was evaluated using the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA). Patients in the combination group were divided into cognitive impairment group (MoCA score ≥ 26 points) and non-cognitive impairment group (MoCA score < 26 points) according to MoCA score. The risk factors of cognitive impairment in patients with ischemic stroke and leukoaraiosis were analyzed. Results:The scores of the MMSE, MoCA, Clock Drawing Test (CDT), Verbal Fluency Test (VFT), and Digit Span Test (DST) in the control group were (28.93 ± 2.70) points, (28.35 ± 2.74) points, (4.69 ± 1.14) points, (4.94 ± 0.42) points, and (14.33 ± 1.66) points respectively. They were (26.92 ± 2.18) points, (25.02 ± 3.52) points, (3.61 ± 1.60) points, (4.77 ± 0.46) points, and (11.73 ± 1.16) points, respectively in the simple ischemic stroke group and (24.91 ± 2.79) points, (20.70 ± 3.06) points, (2.87 ± 1.23) points, (4.07 ± 0.85) points, and (10.82 ± 0.93) points respectively in the combination group. There were significant differences in the scores of the MMSE, MoCA, CDT, VFT, and DST among the three groups ( F = 124.50, 318.50, 93.43, 112.60, 428.60, all P < 0.001). Significant differences in the scores of the MMSE, MoCA, CDT, VFT, and DST were observed between patients with different degrees of leukoaraiosis ( F = 69.09, 102.40, 20.98, 60.90, 57.00, all P < 0.001). Spearman correlation analysis results showed that the scores of the MMSE, MoCA, CDT, VFT, and DST were negatively correlated with the degree of leukoaraiosis ( r = -0.61, -0.69, -0.43, -0.56, -0.44, all P < 0.05). Logistic regression analysis results showed that age, history of smoking and drinking, history of diabetes, history of stroke, and infarct location were the independent risk factors for cognitive impairment in patients with ischemic stroke and leukoaraiosis. Education level was a protective factor against ischemic stroke and leukoaraiosis. Conclusion:The degree of cognitive impairment in patients with ischemic stroke and leukoaraiosis is related to the degree of leukoaraiosis. Age, history of smoking and drinking, history of diabetes, history of stroke, infarction location, and education level are the influential factors of cognitive impairment.

Objective:To illustrate the effectiveness of Micro-Teachermate platform combined with problem-based learning (PBL) in gynecology and obstetrics teaching.Methods:Students from different classes were randomly divided into experimental group and control group, and the general data of the two groups were compared. The two groups respectively adopted Micro-Teachermate platform combined with PBL and traditional lecture teaching methods. The class attendance rate and the scores of relevant chapters in the final examination were analyzed through statistical methods for assessing the enthusiasm of students and teaching quality in class in terms of the same content and the same class hours. A questionnaire survey was conducted to know students' evaluation of this teaching mode, evaluating its teaching effect and satisfaction comprehensively. SPSS 17.0 was used for t test and chi-square test. Results:There was no difference in general information between the two groups. The class attendance rate of the experimental group (274/279) was significantly higher than that of the control group (61/90), while the scores of the relevant chapters in the final exam were also significantly higher ( P<0.01). Questionnaire survey results among 93 students in the experimental group were taken back 93 valid questionnaires (the effective rate, 100%), showing that most of the students were very satisfied with this teaching mode (90/93) and hoped to apply it to more subjects teaching (89/93). Conclusion:The application of Micro-Teachermate platform combined with PBL in gynecology and obstetrics class teaching contributes to improve students' participating enthusiasm and can effectively improve the quality and efficiency of teaching. Most students highly recognize the teaching method, which is worth applying and popularizing.

Objective:To study the clinicopathological features of metaplastic breast cancer (MBC) , and its prognostic factors.Methods:Data of 49 MBC cases and 30 cases of invasive ductal carcinoma (IDC) during the same period as matched control were collected. The immunohistochemistry staining of CK5/6, CK, P63, ER, HER2, Ki67 was performed in all MBC samples.Results:In MBC cases, the median age was 55 years. Median tumor size was 2.5 cm (range, 0.6-19 cm). Fifteen cases were classified as metaplastic carcinoma with heterologous mesenchymal differentiation (8 as matrix-producing carcinoma), 12 as spindle cell carcinoma, 7 as squamous cell carcinoma, 2 as low-grade adenosquamous carcinoma, 2 as fibromatosis-like metaplastic carcinoma, and 11 as mixed metaplastic carcinoma. The 5-, 10-year overall survival rate was 50%, 41%, respectively, lower than those of IDC (76%,63%) (all P<0.05). Lymph node metastasis rate, and expression of ER, PR, HER2 in MBC were lower than those in IDC (all P<0.05). Triple-negative cases in MBC were more than those of IDC ( χ2=26.244, P=0.000). The proliferative index of Ki67 was statistically different between the two groups ( t=2.624, P=0.011). Conclusions:MBC is a rare and heterogenous breast cancer. Compared to IDC, MBCs are usually larger, lower in lymph nodes metastasis, higher in proliferative index of Ki67, more triple-negative, hence with a poorer prognosis.

Objective:To investigate the effect of dexmedetomidine (DEX) on reducing urethral stimulation in patients undergoing laparoscopic gastrointestinal surgery.Methods:From January 2019 to February 2020, 90 patients undergoing elective laparoscopic gastrointestinal surgery under general anesthesia in the Third Affiliated Hospital of Sun Yat-sen University were selected. They were randomly divided into 3 groups: catheterization before induction (group A), catheterization during induction (group B), and catheterization after induction (group C). In group A, patients received general anesthesia after awake catheterization. In group B, intravenous injection of DEX 0.5 μg/kg was pumped for 10 minutes, followed by catheterization and induction. In group C, patients received general anesthesia and then catheterization. Visual analogue scale (VAS) score of urethral stimulation, morphine dosage and the incidence of agitation during resuscitation were recorded. The heart rate and mean arterial pressure of the three groups were compared at the time of entering the room, catheterization, tracheal intubation, entering postanesthesia care unit (PACU), about extubation and 30 minutes after extubation.Results:The fluctuation of blood pressure and heart rate in group B was significantly less than that in group A and group C at the time of extubation and 30 minutes after extubation ( P<0.05). VAS of urethral stimulation in group B [(2.9±0.9)point] was significantly lower than that in group A [(4.4±1.8)point] when catheter was indwelling ( P<0.05). After extubation, VAS in group B [(2.8±1.1)point] was significantly lower than that in group A [(3.2±1.2)point] and C [(5.2±1.8)point] ( P<0.05). The utilization rate of morphine within 24 hours after surgery in group B (10%) was significantly lower than that in the other two groups (40%, 57%), and the incidence of postoperative agitation in group A and B was lower than that in group C within PACU ( P<0.05). The satisfaction of patients in group B (86.7%) was higher than that in group A (70%) and C (46.7%). The satisfaction of PACU personnel in group A (76.7%) and B (80%) was significantly higher than that in group C (43.3%). Conclusions:Sedation with dexmedetomidine during urethral catheterization can reduce urethral stimulation during resuscitation and improve patients' and PACU staffs' satisfaction.

Objective:To illustrate the effectiveness of Teachermate platform combined with problem-based learning (PBL) in gynecology and obstetrics teaching.Methods:Students from different classes were randomly divided into experimental group and control group, and the general data of the two groups were compared. The two groups respectively adopted Teachermate platform combined with PBL and traditional lecture teaching methods. The class attendance rate and the scores of relevant chapters in the final examination were analyzed through statistical methods for assessing the enthusiasm of students and teaching quality in class in terms of the same content and the same class hours. A questionnaire survey was conducted to know students' evaluation of this teaching mode, evaluating its teaching effect and satisfaction comprehensively.Results:There was no difference in general information between the two groups. The class attendance rate of the experimental group (274/279) was significantly higher than that of the control group (61/90) ( P=0.00), while the scores of the relevant chapters in the final exam were also significantly higher ( P=0.00). Questionnaire survey results among 93 students in the experimental group were taken back 93 valid questionnaires (the effective rate, 100%), showing that most of the students were very satisfied with this teaching mode (90/93) and hoped to apply it to more subjects teaching (89/93). Conclusion:The application of Teachermate platform combined with PBL in gynecology and obstetrics class teaching contributes to improve students' participating enthusiasm and can effectively improve the quality and efficiency of teaching. Most students highly recognize the teaching method, which is worth applying and popularizing.