Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.

Objective:This study aims to evaluate the performance of digital PCR (dPCR) detecting multiple and single copies genes of the Epstein-Barr virus (EBV) for nucleic acid quantification and explore their applicability in clinical settings.Methods:Compared the sensitivity, specificity, precision, lower limit of detection (LoD), and linearity for multicopy BamHI-W dPCR and single-copy EBNA1 dPCR systems. Linear regression analysis using the least squares method was employed to evaluate the linearity. Additionally, we analyzed plasma samples from 182 patients with suspected EBV-related diseases between January and July 2022 at the Southern Medical University Southern Hospital, using both dPCR and quantitative PCR (qPCR) for EBV DNA quantification. Linear regression analysis using the least squares method was conducted to assess their quantitative correlation.Results:The dPCR systems for both multicopy and single-copy genes showed excellent linearity ( R 2 values of 0.992 and 0.997, respectively, both P<0.001). The LoD were 188 IU/ml for BamHI-W gene and 358 IU/ml for EBNA1 gene dPCR systems. The logarithmic coefficient of variation ( CV) values for high-concentration samples (1 000 000 IU/ml) were 0.34% and 0.21% for the BamHI-W gene and EBNA1 gene dPCR assays, respectively, while for low-concentration samples (5 000 IU/ml) were 0.98% and 0.64%, respectively. In the detection of seven common clinical infectious pathogens and EBV positive samples, only EBV-positive samples yielded positive signals in the dPCR detection system, with no cross-reaction with other pathogens. In 182 samples, the positive detection rates were 47.80% (87/182) for BamHI-W gene and 35.16% (64/182) for EBNA1 gene dPCR, compared to 43.41% (79/182) for qPCR. Linear correlation analysis with qPCR showed R2 values of 0.837 for BamHI-W gene and 0.763 for EBNA1 gene dPCR (both P<0.001). The BamHI-W gene copy number ranged from 3 to 18 copies per clinical sample, with patient-specific variations. There was a high consistency in viral load trends between the multicopy BamHI-W gene and single-copy EBNA1 gene dPCR systems within individual patients. Conclusions:The dPCR methods detecting EBV multiple and single copies genes showed high sensitivity, specificity, precision, and quantitative accuracy, suitable for clinical sample analysis. The multicopy BamHI-W gene dPCR method notably enhances detection sensitivity and can be used as a supplement to current EBV DNA load detection methods, especially in low-concentration samples. For within-patient EBV DNA monitoring, the multicopy gene method proves more effective, while inter-patient comparisons might necessitate single-copy gene methods or normalize them using the same standard.

Objective:To investigate the etiology, clinical manifestations, clinicopathological features of cystic neutrophil granulomatous mastitis (CNGM).Methods:From Jan 2019 to Dec 2020, 95 CNGM cases diagnosed by biopsy pathology at Chongqing Hospital of Traditional Chinese Medicine and Chongqing Liangping District Hospitol of Traditional Chinese Medicine were reviewed.Results:There were 95 female patients, aged 21 to 50 years, with a median age of 32 years. Laboratory examination showed that 56% (53/95) cases had elevated rheumatoid antibody level, 27 % (26/95) had increased level of serum thyroid antibody, 15% (14/95) had elevated antineutrophil antibody, 35% (33/95) had increased ESR, 38% (36/95) had increased C-reactive protein. The positive rate of Gram-stained bacilli was 82% (78/95). Histology: pyogenic granuloma with lobule of breast as the center, the center of granuloma was cystic vacuole.Immunohistochemistry showed that the inflammatory cells in and around granuloma were mainly CD3 + cells, and CD4 + cells were more than CD8 + cells. Conclusions:The cystic neutrophilie granulo matous mastitis is a rare type of idiopathic granulomatous mastitis. The diagnosis of CNGM is dependent on its specific pathological features.

Receptors, Purinergic ; Signal Transduction/physiology*

Oxidative Phosphorylation ; Acetylglucosamine ; Uridine Diphosphate N-Acetylglucosamine/metabolism*

Objective:To study the risk factors of axillary lymph node metastasis by analyzing the acoustic image characteristics of the automated breast volume scanner (ABVS) of breast cancer masses.Methods:The imaging features of ABVS of 212 patients with breast cancer, unilaterally and singly, confirmed by pathological examination admitted in Hangzhou First People’s Hospital from Jan. 2016 to Dec. 2018 were retrospectively analyzed. There were 83 cases with axillary lymph node metastasis (metastatic group) and 129 cases without (non-metastatic group) . The correlation of clinical and the imaging features of ABVS with axillary lymph node metastasis was analyzed using univariate analysis and multivariate logistic regression. ROC curve was used to analyze the cut-off value of the maximum diameter of the mass in predicting the breast cancer axillary lymph node metastasis. The sensitivity, specificity, positive predictive value, and negative predictive value of each risk factor were analyzed for predicting breast cancer axillary lymph node metastasis.Results:The retraction phenomenon and micro-calcification of breast cancer in the metastatic group (60.2%, 65.1%) were higher than those in the non-metastatic group (43.4%,37.2%) ( P=0.017 vs P<0.001) . The maximum diameter of the breast cancer in the metastatic group was bigger than in the non-metastatic group ( Z=2.18, P=0.029) . Multivariate logistic regression analysis showed that the micro-calcification of breast cancer ( OR=2.522, P=0.003) was an independent predictor of lymph node metastasis in breast cancer. The area under the curves and the cut-off value of the maximum diameter of the mass in predicting the breast cancer axillary lymph node metastasis were 0.589 and 2.85 cm. The sensitivity was 34.9%, the specificity was 82.9%, the positive predictive value was 56.9%, and the negative predictive value was 66.5%. The sensitivities of micro-calcification and retraction phenomenon to predict the occurrence of axillary lymph node metastasis in breast cancer patients were 65.1% and 60.2%, specificities were 62.8% and 56.6%, positive predictive values were 52.9% and 47.2%, and negative predictive values were 73.6% and 68.9%. Conclusion:The study suggests that the maximum diameter, micro-calcification, and retraction phenomenon of masses are associated with the occurrence of the axillary lymph node metastasis in breast cancer.

Objective To investigate the value of the automated breast volume scanning (ABVS) combined with virtual touch tissue imaging quantification (VTIQ) in the differential diagnosis of benign and malignant breast lesion.Methods Five hundred and seven patients with a total of 675 breast nodules were detected using ABVS and VTIQ technique.Of them,coronal plane imaging,SWVmax,SWVmin,and SWVmean (in m/s) for each nodule was acquired three days before operation or core needle biopsy.According to pathological results,the receiver operating characteristic (ROC) curve analysis was performed to evaluate the utility of the ABVS alone (retraction phenomenon on coronal plane),VTIQ alone and their combination in the diagnosis of breast lesions.Results Among all nodules,504 lesions were benign,and 171 were malignant.The rate of retraction phenomenon on coronal plane in malignant lesions was significantly higher than that in benign lesions (x2=279.89,P ＜ 0.001).The value of SWVmax,SWVmin,SWVmean (6.79± 1.71 m/s,5.03 ± 1.24 m/s,5.74± 1.36 m/s) in malignant nodules were higher than that of benign nodules [(3.41 ±1.51) m/s,(2.46± 0.87) m/s,(2.65 ± 1.23) m/s],the differences were statistically significant (t=32.43,33.85,26.77,all P ＜ 0.001).The AUC of malignant nodules for SWV (maximum,minimum,and average) were 0.922,0.934 and 0.937,respectively.With cut-off value of 4.045 m/s,SWVmean showed the best diagnostic performance.The sensitivity,specificity and accuracy of the retraction phenomenon on coronal plane,SWVmean and their combination in diagnosing malignant breast lesion were (39.65％,94.39％,95.78％),(97.50％,94.39％,95.53％) and (82.84％,93.51％,94.25％),respectively.And the SWVmean showed significant higher sensitivity than that of retraction phenomenon.The diagnostic capacity significantly improved when the two approaches were combined.Conclusion Both the retraction phenomenon on coronal plane and VTIQ had high diagnostic value,combining the two methods can improve the differential diagnosis ability for breast lesions.

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .