Objective:To carry out prenatal diagnosis for a fetus with Meckel syndrome(MKS) and explore its genetic basis.Methods:A pregnant woman presented at Suzhou Municipal Hospital in February 2018 was selected as the study subject. Clinical data was collected. Muscle tissue sample from the abortus and peripheral blood samples from the couple were collected. Genomic DNA was extracted and subjected to chromosomal microarray analysis (CMA) and whole exome sequencing. Candidate variant was verified by Sanger sequencing.Results:The fetus was found to have microcephaly, oligohydramnios, polycystic kidneys and banana-shaped cerebellum at 18 weeks of gestation. After induction of labor, it was found to have encephalocele, renal cysts and polydactyly. CMA has found no abnormality. Whole exome sequencing revealed novel compound heterozygous variants c. 296delA (p.Lys99SerfsTer6) and c. 1243G>A (p.Val415Met) in the TMEM67 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 296delA variant was predicted to be pathogenic (PVS1+ PM2_Supporting+ PP4), whilst the c. 1243G>A variant was predicted to be likely pathogenic (PM2_Supporting+ PM3+ PP3_Moderate+ PP4). Conclusion:The c. 296delA and c. 1243G>A compound heterozygous variants of the TMEM67 gene probably underlay the MKS in this fetus.

Objective:To investigate the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of SRF-rearranged cellular perivascular myoid tumor.Methods:Two cases of SRF-rearranged cellular perivascular myoid tumor diagnosed in the Department of Pathology, Fudan University Shanghai Cancer Center from October 2021 to March 2022 were collected. Immunohistochemical staining, fluorescence in-situ hybridization (FISH) and next-generation sequencing (NGS) were performed, and the literature was reviewed.Results:Case 1, a 3-month-old boy presented with a painless tumor of the scalp, measuring about 2 cm in diameter. Case 2, a 3-year-old girl complained with a painless tumor of the knee, measuring approximately 1.5 cm in diameter. Microscopically, the tumor had a clear boundary and showed multinodular growth. The tumor was mainly composed of spindle cells arranged in long intersecting fascicles associated with thin, slit-like or branching ectatic vessels, focally forming hemangiopericytoma-like appearance. The tumor cells were abundant, but there was no obvious atypia. Mitotic figures (3-4/10 HPF) were noted. H-caldesmon and SMA were positive in both cases. Case 1 showed diffuse and strong positivity for Desmin, and focally for CKpan. Ki-67 proliferation index was 20% and 30%, respectively. FISH displayed NCOA2 gene translocation in case 1 and the RELA gene translocation in case 2. NGS detected the SRF-NCOA2 gene fusion in case 1 and the SRF-RELA gene fusion in case 2. Both patients underwent local excisions. During the follow-up of 5-14 months, case 1 had no local recurrence, while case 2 developed local recurrence 1 year post operatively.Conclusions:SRF-rearranged cellular perivascular myoid tumor is a novel variant of perivascular cell tumor, which tends to occur in children and adolescents. The tumor forms a broad morphologic spectrum ranging from a pericytic pattern to a myoid pattern, and include hybrid tumors with a mixture of pericytic and myoid patterns. Due to its diffuse hypercellularity and increased mitotic figures and smooth muscle-like immunophenotype, the tumor is easy to be misdiagnosed as myogenic sarcomas. The tumor usually pursues a benign clinical course and rare cases may locally recur.

Purpose To investigate the effect of MYD88 gene overexpression on the proliferation and apoptosis of human diffuse large B cell lymphoma(DLBCL)cells,and to prelimi-narily explore the mechanism of MYD88 gene action.Methods PEGFP-C2-MYD88 overexpressing MYD88 L265P gene was transfected into DLBCL cells by plasmid transfection.The exper-iment was divided into blank control group,negative control group and MYD88 L265P overexpression group.The fluores-cence expression of MYD88 L265P after overexpression was ob-served under inverted fluorescence microscope.RT-PCR and Western blot were used to detect the mRNA and protein expres-sion of MYD88 L265P,IRAK4,NF-κB and BCL2 in DLBCL cells before and after overexpression of MYD88 L265.CCK8 method was used to detect DLBCL cells proliferation and Ho-echst staining was used to detect DLBCL cells apoptosis.Re-sults After overexpression of MYD88 L265P,compared with the blank control group(0.670 4±0.017 5)and the negative control group(0.715 3±0.019 6),the MYD88L265P overex-pression group(1.157 2±0.010 2)increased significantly,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(0.69 ±0.04)and the negative control group(0.81±0.07),the MYD88L265P overexpression group(0.48±0.05)was signifi-cantly decreased,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(mRNA:1.0158±0.0115,0.987 3±0.010 2,1.007 6±0.015 3,protein:0.183 4±0.058 9,0.096 8± 0.015 7,0.147 5±0.0418)and negative control group(mR-NA:0.9132±0.0098,1.0032±0.0156,0.9327± 0.011 2,protein:0.187 9±0.042 3,0.088 9±0.0513,0.134 8±0.050 1),the mRNA(3.243 2±0.013 6,2.976 6 ±0.0213,1.585 9±0.019 8)and protein expressions(0.452 7±0.052 4,0.218 9±0.047 5,0.301 4±0.059 8)of IRAK4,NF-κB and anti-apoptosis protein BCL2 in MYD88L265P overexpression group were significantly increased,which was statistically significant(all P＜0.05).Conclusion After overexpression of MYD88 L265P,the apoptosis rate of DLBCL cells decreased and the cell proliferation rate increased.The mechanism may be related to the mutation of MYD88 L265P gene,activation and amplification of NF-κB pathway,and pro-motion of the overexpression of antiapoptotic protein BCL2.

Based on the teaching advantages of "Internet +", we integrated the ATDE(asking, thinking, doing, evaluation) teaching model to explore the online teaching mode of neurology during the COVID-19 epidemic, and chose the ischemic cerebrovascular disease as a representative case of specific neurological diseases. The practical ATDE teaching in the teaching of neurology was elaborated by introducing, inspiring doubt, setting suspense, and raising questions, and leading students gradually to think deeply. Finally, the innovation and thinking of the online teaching of neurology brought by "Internet +ATDE" teaching model are discussed.

OBJECTIVE：To simultaneo usly determine the contents of atractylenolide Ⅱ ，β-eudesmol，atractyloxin and atractylone in Atractylodes chinensis ，and to evaluate the quality of A. chinensis with different growth years combined with color difference principle. METHODS ：HPLC method was adopted. The determination performed on Agilent Eclipse XDB-C 18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid （gradient elution ）at the flow rate of 1.0 mL/min；the detection wavelengths were set as 208 nm（atractylenolide Ⅱ，β-eudesmol），340 nm（atractyloxin）and 220 nm（atractylone）；the sample size was 15 μ L. Using atractyloxin as reference，QAMS was adopted to establish relative correction factors （RCFs） of atractylenolideⅡ，β-eudesmol and atractylone ；the content of each component in A. chinensis with different growth years were calculated. The contents of above 4 components were determined by external standard method and then compared with the results of QAMS. The color difference values of A. chinensis powder were measured based on color difference principle. The correlation analysis of above 4 components content with color was carried out by Pearson correlation analysis. RESULTS ：The separation degree of atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis was higher than 1.5. The linear range were 1.01-10.10，3.30-33.00，4.40-44.00，5.34-53.40 μg/mL，respectively. RSDs of precision ，reproducibility and stability tests were all lower than 2％，while the average recovery rates were 101.34％-104.67％（RSD＜1.5％，n＝6）. Using atractyloxin as reference ， RCFs of atractylenolide Ⅱ，β-eudesmol and atractylone were 3.896 7，5.928 2，9.727 9，with RSD of 0.35％，2.89％，0.36％ （n＝6），respectively. Relative deviation of 3 components （except for atractyloxin ） in 24 batches of A. chinensis ranged 0.03％-1.45％ between QAMS and external standard method ，which indicated that the results of two methods were consistent ，and the content of each component increased with the increase of growth years. Atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis had significant negative correlation with its color shade （L*），total color difference （E*ab）（P＜0.01）， and significant positive correlation with color red-green direction （a*）， color yellow-blue direction （b*）（P＜0.01）. CONCLUSIONS：The established HPLC-QAMS method can be used for the determination of atractylenolide Ⅱ，β-eudesmol， atractyloxin and atractylone in A. chinensis . The longer the growth period is ，the higher each component content is. The color of A. chinensis is closely related to the content of each component ，and the content of effective components is higher in A. chinensis with dark yellowish brown color.

OBJECTIVE： To establish the method for the content determination of gentiopicrin and loganic acid in Gentiana scabra， and to investigate the correlation of their contents with appearance traits and quality gradation criterion. METHODS： HPLC method was adopted. The determination was performed on Ascentis Express C18 column with mobile phase consisted of 0.1％ phosphoric acid-acetonitrile （gradient elution） at the flow rate of 0.4 mL/min. The column temperature was 30 ℃， and detection wavelength was set at 240 nm. The sample size was 1 μL. Taking the length， number and diameter of fibrous roots as indexes， the appearance and morphological characteristics of G. scabra were studied. The relationship of gentiopicrin and loganic acid content with the appearance property of medicinal material was analyzed by SPSS 21.0 software. k-mean clustering analysis was carried out by using SPSS 21.0 software， and gradation standard for G. scabra was established preliminarily. RESULTS： The linear range of gentiopicrin and loganic acid were 0.5-3.0 μg/mL （r＝0.999 9） and 0.05-0.50 μg/mL （r＝0.999 9）. The limit of quantification of gentiopicrin and loganic acid were 0.295， 0.289 μg/mL； the detection limit were 0.082， 0.081 μg/mL； RSDs of precision， stability， repeatability tests were all lower than 2％； the recovery rates were 97.56％- 102.23％ （RSD＝1.56％， n＝6） and 97.58％-102.67％ （RSD＝1.86％， n＝6）. Correlation results showed that there was a significant positive correlation of the length of G. scabra， the number of roots， root diameter， with the contents of gentiopicrin and loganic acid. The order of affecting content was the number of roots ＞length ＞root diameter. k-means clustering analysis showed that 54 batches of G. scabra was divided into two categories； S4-S6，S13，S17-S23，S25，S28，S31-S34 were clustered into a category； S1-S3， S7-S12， S14-S16， S24， S26，S27，S29， S30，S35-S54 were clustered into the other category. The results of gradation showed that 54 batches of G. scabra could be divided into two grades， and the results were consistent with the cluster analysis. CONCLUSIONS： Established method is simple and stable， and can be used for simultaneous determination of gentiopicrin and loganic acid in G. scabra. The more fibrous roots， the longer the length， the thicker the root， the higher the content of gentiopicroside and loganic acid， the better the quality of G. scabra.

Objective To compare and analyze the short-term effect of play-based communication and behavior intervention (PCBI) and applied behavioral analysis (ABA) on the treatment of toddlers with autism spectrum disorder (ASD).Methods Seventy-four ASD toddlers aged from 19 to 30 months were recruited in Nanjing Brain Hospital Affiliated to Nanjing Medical University from November 2017 to May 2018.The toddlers who participated in this study were randomly assigned into PCBI group and ABA group,then they were intervened weekly by PCBI or ABA for a total of 12 weeks.Portage Early Development Checklist and autism treatment evaluation checklist (ATEC) were used to estimate the toddlers' developmental level and the treatment efficacy respectively.The t-test was used to reveal whether there was significant difference between the 2 groups before and after intervention.Results (1) Compared with the ABA group,there was a significant increase in cognitive scores (△PCBI =9.03 scores,△ABA =4.27 scores,t =3.997) and a significant decrease in social behavior scores (△PCBI =8.87 scores,△ABA =16.91 scores,t =-4.022) of the Portage Early Development Checklist after 12 weeks of intervention in the PCBI group,and there were statistically significant differences(all P ＜ 0.001);while the total score and the scores of language,movement,and self-care subscale of Portage Early Development Checklist were also increased after PCBI intervention but without significant differences (all P ＞ 0.05).(2)Compared with the ABA group,after 12 weeks of PC BI intervention,the scores of social contact,perception,behavior of ATEC were decreased,but the difference was not significant (P ＞ 0.05);the total score of ATEC scale(△PCBI total =14.89 scores,△ABA total =22.22 scores,t =2.209)and the scores of language subscale(△PCBI language =2.89 scores,△ ABA language =6.43 scores,t =2.515) were decreased significantly,and there were statistically significant differences (all P ＜ 0.05).Conclusions After 12 weeks of the very early intervention of PCBI,the ASD toddlers all improved in clinical symptoms and developmental level.Compared with ABA intervention,PCBI very early intervention with parental guidance was comparable in short term efficacy,and PCBI was not taking up as much medical rehabilitation resources as ABA.

Objective: Children with autism spectrum disorder(ASD) who participated in the " Play-based Communication and Behavior Intervention(PCBI)" were studied in order to evaluate the impact of caregiver factors on the efficacy of the PCBI very early intervention model systematically. Methods: Seventy-six children with ASD aged 8-30 months were recruited in this study at Brain Hospital Affiliated to Nanjing Medical University form December 2017 to June 2018, and they were trained for PCBI very early intervention.The Autism Treatment Evaluation Checklist (ATEC) and the Gesell Developmental Schedules were used to evaluate the efficacy of the PCBI very early intervention.Parenting Stress Index Short Form, General Self-Efficacy Scale and the self-developed Caregiver Training Course Evaluation Scale and the Homework Completion Level Scale were used to assess the level of parental stress, self-efficacy, curriculum satisfaction and acceptance.A pair of sample t-test was used to analyze the ASD symptoms, developmental level and caregiver factors of ASD children before and after intervention.Multivariate regression analysis was used to analyze the caregiver factors and the effects of PCBI very early intervention. Results: (1) At the end of 12 weeks of PCBI very early intervention, the score of the ATEC (17.36±15.98) was lower than that before intervention (78.43±22.84), and the difference was statistically significant(P<0.05); the adaptive developmental quotient(5.90±15.45), the fine action developmental quotient(5.13±19.89), the language developmental quotient (12.43±20.60) and the personal social developmental quotient(4.79±16.02) scores of the Gesell Developmental Schedules were higher than those before intervention (73.08±20.54, 77.33±23.63, 52.24±19.12, 71.79±20.81), and the differences were statistically significant (all P<0.05). (2) At the end of PCBI very early intervention, there was a significant decrease in the scores of Parenting Stress Index Short Form, and the differences were statistically significant (4.84±14.69 vs.94.05±29.67, P<0.05). (3) Acceptance of the intervention model, and mastery of the skills in the caregiver factors had a positive predictive effect on the effect of PCBI very early intervention (t=3.068, 10.468, all P<0.05). Conclusions PCBI very early intervention can effectively reduce the parenting pressure of the caregivers, and in the PCBI very early intervention, the better the mastery of skills and the completion of homework, the better the efficacy of intervention.

Objective: To explore the application values of prenatal ultrasound, vascular cast in the diagnosis of fetal aortic arch and its branches anomalies and to analyze the genetic characteristics by gene detection. Methods: Twenty-two cases of the vascular cast specimens of the fetal aortic arch and its branches anomalies were analyzed and studied by comparing with their prenatal ultrasonography. Then the characteristics of each type of fetal aortic arch and its branches anomalies, the missed diagnosis and misdiagnosis were summarized and the results of their gene detection were also analyzed. Results: The 22 cases of fetal aortic arch and its branches anomalies were as follows: 2 cases of double aortic arch showed the ascending aorta was divided into two branches after converging as the descending aorta. Three cases were left aortic arch with aberrant right subclavian artery. Twelve cases were right aortic arch: 8 cases were right aortic arch with mirror-image branching, 3 cases were right aortic arch with aberrant left subclavian artery, 1 case was right aortic arch with isolated left subclavian artery. Of the 8 right aortic arch with mirror-image branching, 3 cases of left arterial duct showed the vertical walking between the fusion site of the left innominate artery and the pulmonary artery. Right aortic arch with aberrant left subclavian artery with arterial duct showed "U" shaped vascular ring. Five cases were other types, including 2 cases of the coarctation of aortic arch, 1 case of interrupted aortic arch, 1 case of pulmonary artery sling, and 1 case of abnormal origin of right pulmonary artery. The ultrasonic missed diagnosis were the 6 deformities: 3 cases of arterial duct and 3 cases of aberrant subclavian artery. The ultrasonic misdiagnosis were the 5 deformities: 2 cases of arterial duct location, 1 case of aberrant subclavian artery, 1 case of isolated left subclavian artery, and 1 case of the coarctation of aortic arch. Genetic test results: In the 18 cases of the genetic detection, 2 cases were positive, 1 case was 22q11.2 microdeletion syndrome and 1 case was carrying KMT2D gene variant. Conclusions There are various kinds of fetal aortic arch and its branches anomalies, which are often associated with intracardiac malformations and venous branches variation. And prenatal ultrasound is of great value in diagnosing them.Vascular cast can visually display their characteristic changes, which is helpful to improve the differential diagnosis of the different aortic arch and its branches anomalies. The detailed genetic detection can improve the further understanding of its etiology.

Objective To explore the effect of blood oxygen saturation on the hemodynamics in fetuses with complete transposition of the great artery ( CTGA ) without outflow tract obstruction in the second trimester of pregnancy . Methods The brain biometry and hemodynamics parameters were retrospectively analyzed in 38 fetuses with CTGA from multiple medical centers .All the fetuses with CTGA were divided into two groups ,ventricular septal defect ( VSD) group( 14 cases) and no VSD group( 24 cases) according to the VSD . Twenty-four gestational age (GA)-matched fetuses were chosed as control group . The biparietal diameter (BPD) and head circumference ( HC) ,pulsatility indexes ( PI) in the middle cerebral artery (MCA) and the umbilical artery ,the cerebroplacental ratio (CPR) were compared among different groups . Results There were no significant differences in GA ,BPD ,HC ,MCA-PI ,UA-PI and CPR among control group ,CTGA with VSD group and CTGA without VSD group in the second trimester of pregnancy ( P ＞0 .05) . Conclusions The altered oxygen saturation of the cerebral circulation in CTGA does not affect the brain biometry and hemodynamics in fetuses in the mid-gestational age .