OBJECTIVE: To assess the clinical value of digital PCR (dPCR) for the prenatal diagnosis of common fetal aneuploidies. METHODS: A dPCR-based assay was developed for detecting trisomies 21, 18, and 13. A retrospective analysis was carried out on 173 amniotic fluid samples collected by the Prenatal Diagnosis Center of Taizhou Hospital between January 2017 and December 2023. By using chromosomal karyotyping as the gold standard, the diagnostic performance of the multiplex dPCR system was evaluated in a double-blind manner. This study has been approved by the Ethics Committee of Taizhou Hospital (Ethics No. K20250339). RESULTS: Chromosomal karyotyping has identified 59 cases of trisomy 21, 5 cases of trisomy 18, 2 cases of trisomy 13, 6 cases with chromosomal structural abnormalities or mosaicisms, and 101 cases with a normal karyotype. The dPCR results (Z-score cutoff = 4.0, CI = 99.997%) showed full concordance with karyotyping (sensitivity = 100%, specificity = 100%, Kappa = 1). Among the 6 structurally abnormal or mosaicism samples, dPCR has accurately detected 4 cases, but mis-classified 2 cases of trisomy 21 with very low-level mosaicisms (3.3%, 6.9%, respectively) as normal. CONCLUSION The established multiplex dPCR system demonstrated high diagnostic accuracy for common chromosomal aneuploidies, with results available within 24 hours. It can serve as an efficient supplementary tool to conventional chromosomal karyotyping, providing reliable support for time-sensitive clinical decision-making in prenatal diagnosis.
Humans ; Female ; Pregnancy ; Aneuploidy ; Prenatal Diagnosis/methods* ; Karyotyping ; Retrospective Studies ; Polymerase Chain Reaction/methods* ; Chromosome Disorders/genetics* ; Adult ; Trisomy 13 Syndrome/diagnosis* ; Trisomy 18 Syndrome/genetics* ; Down Syndrome/genetics*

OBJECTIVE: To assess the clinical value of non-invasive prenatal testing (NIPT) for identifying maternal malignant tumors. METHODS: A retrospective analysis was carried out on pregnant women undergoing Non-invasive prenatal testing (NIPT) at Taizhou Hospital in Zhejiang Province from January 2018 to December 2022. The criteria included maternal copy number variations for at least two chromosomes. Clinical follow-up data were obtained for the high-risk population of maternal malignant tumors through telephone follow-up and review of electronic medical records. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: K20250339). RESULTS: Among 45 141 NIPT samples, 6 (0.013%) were suggested to have maternal malignant tumors. Follow-up information was available for 5 patients (83.3%). Two cases were diagnosed with maternal malignant tumors, including 1 myelodysplastic syndrome and 1 pelvic malignant tumor. Two cases were found to have multiple uterine fibroids and 1 was lost during follow-up. CONCLUSION The abnormal copy number indicated by NIPT may serve as an early signal for maternal malignant tumors. To establish a systematic follow-up protocol and multidisciplinary collaboration are conducive to achieving early diagnosis of tumors and improving the prognosis of patients. Based on the results of this study, it is recommended that for pregnant women with unexplained copy number variations and suspected maternal tumors by NIPT, targeted tumor screening program should be implemented to optimize their clinical management.
Humans ; Female ; Pregnancy ; Adult ; Retrospective Studies ; DNA Copy Number Variations/genetics* ; Follow-Up Studies ; Neoplasms/diagnosis* ; Noninvasive Prenatal Testing/methods* ; Pregnancy Complications, Neoplastic/diagnosis* ; Prenatal Diagnosis/methods*

OBJECTIVE: To investigate the clinical characteristics and genetic etiology of a male with a normal phenotype and SRY gene-positive 46,XX/46,XY tetrazoospermia chimerism. METHODS: A male patient with an abnormal peripheral blood chromosomal karyotype detected at the Infertility Center of Taizhou Hospital of Zhejiang Province on December 2, 2013 was selected as the study subject. Peripheral venous blood samples were collected from the proband and his family members, together with a semen sample from the proband. Chromosomal karyotype analysis, red blood cell blood group identification, chromosomal microarray analysis (CMA), fluorescence in situ hybridization (FISH), sex-determining region Y (SRY) gene detection, and short tandem repeat (STR) microsatellite marker analysis were performed on the peripheral venous blood sample from the proband. Routine semen analysis, sperm FISH, and STR testing were also conducted. STR verification was performed on both parents. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: k20201009). RESULTS: The proband, a 37-year-old male, had normal secondary sexual characteristics and external genitalia development. The chromosomal karyotype of his peripheral blood sample was 46,XX[94]/46,XY[6]. ABO blood group typing was positive for Rh(D) type O and negative for Rh(D) type A, indicating the presence of two red blood cell populations. CMA result was arr[GRCh37](1-22)×2,(XX)×1. Autosomal and X chromosome SNP genotypes were BB-BB, AB-AB, and AA-AA, making it impossible to identify homozygous/heterozygous chimerism. FISH detection of interphase nuclei showed nuc ish XX[92]/XY[8]. Testing of the SRY gene was positive. STR analysis showed a single X peak (no Y peak) at the AMEL locus, 10/12 at the Penta D locus, and no third allele at other loci. Routine semen analysis were normal. Sperm FISH detection showed haploid nuclei nuc ish X[53]/Y[47]. Sperm STR analysis revealed an X/Y bimodal distribution at the AMEL locus and a 9/14 distribution at the Penta D locus, with no third allele observed at other loci. Above results suggested that the proband's blood and germ cell lines had originated from a heterozygous chimera formed by the fusion of two different zygotes. CONCLUSION Combined genetic techniques confirmed that the proband's peripheral blood AMEL genotype is X/X, while the sperm is X/Y. The Penta D locus showed a bi-allelic heterozygous pattern of 10/12 in the peripheral blood sample and 9/14 in the sperm sample, suggesting that the proband is a tetrazygotic chimera resulted from the fusion of 46,XX/46,XY zygotes.
Humans ; Male ; Adult ; Chimerism ; Microsatellite Repeats ; Sex-Determining Region Y Protein/genetics* ; Phenotype ; Genes, sry ; In Situ Hybridization, Fluorescence ; Karyotyping

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.

Objective:To investigate the safety and effectiveness of emergency one-stop hybrid operation in ruptured and hemorrhagic high-grade brain arteriovenous malformations (BAVMs).Methods:Twelve patients with ruptured and hemorrhagic high-grade BAVMs who underwent emergency one-stop hybrid operation were chosen. The clinical data and efficacy of these patients were analyzed retrospectively.Results:All patients completed surgery successfully. Embolization combined with BAVMs resection was performed in 4 patients, BAVMs resection under digital subtraction angiography (DSA) was performed in 4 patients, embolization combined with balloon-assisted BAVMs resection was performed in 1 patient, balloon-assisted BAVMs resection under DSA was performed in 1 patient, embolization combined with BAVMs resection and aneurysm clipping was performed in 1 patient, and BAVMs resection combined with aneurysm clipping under DSA was performed in 1 patient. Intraoperative DSA showed residual vascular malformation in 1 patient and complete resection in other 11 patients. No surgical complications occurred. Two patients presented symptoms of delayed epilepsy after discharge. Six months after operation, 8 patients had good prognosis (Glasgow Outcome Scale [GOS] scores≥4), 4 had poor prognosis (GOS scores of 1-3), and no death occurred. CTA or DSA in all patients 6 months after operation found no residual or recurrent vascular lesions.Conclusion:Emergency one-stop hybrid operation has high complete occlusion rate and low surgical complication incidence in ruptured and hemorrhagic high-grade BAVMs, which is worthy of clinical application.

By measuring the relative expression level of miR-1825 in serum of pre-operative and post-operative patients with breast cancer and healthy subjects, the clincal value of miR-1825 for pre-operative and post-operative breast cancer patients was evaluated.The serum of pre-operative breast cancer patients( n=92), post-operative breast cancer patients( n=64) and healthy subjects( n=60) were collected from General Hospital of Southern Theatre Command of PLA from October 2018 to March 2021. Real-time quantitative PCR was used to detect the relative expression of miR-1825 in the serum of breast cancer patients and healthy controls. The clinicopathological data were used to analyze the correlation between the expression level of miR-1825 and serum tumor markers level. The receiver operating characteristic curve (ROC) was used to evaluate the diagnosis value of breast cancer with miR-1825, CA15-3. Mann-Whitney U test was used for comparisons between two groups,and Kruskal-Wallis H test was used for multiple group comparisons. The correlation between miR-1825 and CEA, CA15-3, CA-125 expression were analyzed using Spearman correlation test.The relative expression level of miR-1825 in serum of pre-operative patients with breast cancer 1.290(0.705, 1.793) was significantly higher than that of healthy controls 0.18(-0.876, 0.725), but decreased after surgery and chemotherapy -0.080(-0474, 0.405). The analysis of clinicopathological characteristics found that the expression level of miR-1825 was higher in patients with stage Ⅲ-Ⅳ, low degree of tissue differentiation, and tumor larger than 2 cm[stageⅠ-Ⅱ:0.975(0.458, 1.380), stageⅢ-Ⅳ: 1.955(1.663, 2.535), U=98.000, P<0.001;low degree of tissue differentiation:1.685(1.448, 2.143), high/medium degree of tissue differentiation:0.700(0.395, 0.898), U=15.500, P<0.001; tumor smaller than 2 cm:0.935(0.438, 1.370), tumor larger than 2 cm:1.915(1.580, 2.288), U=215.500, P<0.001].Spearman analysis result showed that the expression of serum miR-1825 in breast cancer patients was linearly correlated with the expression of CEA ( r=0.274, P=0.008) and CA15-3 ( r=0.587, P<0.001); ROC curve result showed that miR-1825 was able to distinguish preoperative breast cancer patients from healthy people and postoperative patients. When using one biomarker to discriminate pre-operation and post-operation patients,miR-1825 had the best diagnostic efficiency,with an area under the ROC curve(AUC) of 0.914(95 %CI: 0.872-0.956). miR-1825 may become a potential serum marker for the diagnosis of breast cancer and monitoring of therapeutic efficacy.

By measuring the relative expression level of miR-1825 in serum of pre-operative and post-operative patients with breast cancer and healthy subjects, the clincal value of miR-1825 for pre-operative and post-operative breast cancer patients was evaluated.The serum of pre-operative breast cancer patients( n=92), post-operative breast cancer patients( n=64) and healthy subjects( n=60) were collected from General Hospital of Southern Theatre Command of PLA from October 2018 to March 2021. Real-time quantitative PCR was used to detect the relative expression of miR-1825 in the serum of breast cancer patients and healthy controls. The clinicopathological data were used to analyze the correlation between the expression level of miR-1825 and serum tumor markers level. The receiver operating characteristic curve (ROC) was used to evaluate the diagnosis value of breast cancer with miR-1825, CA15-3. Mann-Whitney U test was used for comparisons between two groups,and Kruskal-Wallis H test was used for multiple group comparisons. The correlation between miR-1825 and CEA, CA15-3, CA-125 expression were analyzed using Spearman correlation test.The relative expression level of miR-1825 in serum of pre-operative patients with breast cancer 1.290(0.705, 1.793) was significantly higher than that of healthy controls 0.18(-0.876, 0.725), but decreased after surgery and chemotherapy -0.080(-0474, 0.405). The analysis of clinicopathological characteristics found that the expression level of miR-1825 was higher in patients with stage Ⅲ-Ⅳ, low degree of tissue differentiation, and tumor larger than 2 cm[stageⅠ-Ⅱ:0.975(0.458, 1.380), stageⅢ-Ⅳ: 1.955(1.663, 2.535), U=98.000, P<0.001;low degree of tissue differentiation:1.685(1.448, 2.143), high/medium degree of tissue differentiation:0.700(0.395, 0.898), U=15.500, P<0.001; tumor smaller than 2 cm:0.935(0.438, 1.370), tumor larger than 2 cm:1.915(1.580, 2.288), U=215.500, P<0.001].Spearman analysis result showed that the expression of serum miR-1825 in breast cancer patients was linearly correlated with the expression of CEA ( r=0.274, P=0.008) and CA15-3 ( r=0.587, P<0.001); ROC curve result showed that miR-1825 was able to distinguish preoperative breast cancer patients from healthy people and postoperative patients. When using one biomarker to discriminate pre-operation and post-operation patients,miR-1825 had the best diagnostic efficiency,with an area under the ROC curve(AUC) of 0.914(95 %CI: 0.872-0.956). miR-1825 may become a potential serum marker for the diagnosis of breast cancer and monitoring of therapeutic efficacy.

Since the outbreak of COVID-19 pandemic, the detection capability has been improving and the detection techniques have been evolving with innovations. qRT- PCR and mNGS, which represent the current mainstay diagnostic technologies, play key roles in disease diagnosis and monitoring of virus variation. The detection technologies based on serum and plasma IgM and IgG antibodies are important for auxiliary diagnosis. RT-LAMP is highly specific for a diagnostic purpose. Digital PCR could quantitatively detect nucleic acid and SHERLOCK has a higher sensitivity. These techniques all have great potential for future development and application for pathogen detection. In this review the authors summarize the basic rationales, technical characteristics and the current application of the SARS-CoV-2 detection techniques.
Antibodies, Viral ; Betacoronavirus ; Clinical Laboratory Techniques ; Coronavirus Infections ; diagnosis ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Pandemics ; Pneumonia, Viral ; diagnosis

Objective    To summarize the experience of minimally invasive anterior mediastinal tumor resection in our center, and compare the Da Vinci robotic and video-assisted thoracoscopic approaches in the treatment of mediastinal tumor. Methods    A retrospective cohort study was conducted to continuously enroll 102 patients who underwent minimally invasive mediastinal tumor resection between September 2014 and November 2019 by the single medical group in our department. They were divided into two groups: a robotic group (n=47, 23 males and 24 females, average age of 52 years) and a thoracoscopic group (n=55, 29 males and 26 females, average age of 53 years). The operation time, intraoperative blood loss, postoperative thoracic drainage volume, postoperative thoracic drainage time, postoperative hospital stay, hospitalization expense and other clinical data of two groups were compared and analyzed. Results    All the patients successfully completed the surgery and recovered from hospital, with no perioperative death. Myasthenia gravis occurred in 4 patients of the robotic group and 5 of the thoracoscopic group. The tumor size was 2.5 (0.8-8.7) cm in the robotic group and 3.0 (0.8-7.7) cm in the thoracoscopic group. Operation time was 62 (30-132) min in the robotic group and 60 (29-118) min in the thoracoscopic group. Intraoperative bleeding volume was 20 (2-50) mL in the robotic group and 20 (5-100) mL in the thoracoscopic group. The postoperative drainage volume was 240 (20-14 130) mL in the robotic group and 295 (20-1 070) mL in the thoracoscopic group. The postoperative drainage time was 2 (1-15) days in the robotic group and 2 (1-5) days in the thoracoscopic group. There was no significant difference between the two groups in the above parameters and postoperative complications (P>0.05). The postoperative hospital stay were 3 (2-18) days in the robotic group and 4 (2-14) in the thoracoscopic group (P=0.014). The hospitalization cost was 67 489 (26 486-89 570) yuan in the robotic group and 27 917 (16 817-67 603) yuan in the thoracoscopic group (P=0.000). Conclusion    Compared with the video-assisted thoracoscopic surgery, Da Vinci robot-assisted surgery owns the same efficacy and safety in the treatment of mediastinal tumor, with shorter postoperative hospital stay, but higher cost.

BACKGROUND: Primary lung cancer and metastatic lung cancer are common malignant tumors of the lung and are the main cause of cancer-related death. Advanced lung cancer and lung metastatic cancer are mainly treated by systemic therapy, and local treatment is also an effective treatment for the refractory or recurrent lesions in the lungs after systemic treatment. ¹²⁵I radioactive particle implantation, as an efficient conformal radiotherapy, has a certain control effect on localized lung cancer. The purpose of this study was to investigate the clinical efficacy and safety of computed tomography (CT) guided percutaneous puncture of lung tissue with ¹²⁵I radioactive particle implantation in the treatment of advanced lung cancer and lung metastatic cancer. METHODS: Continuouslycollectedthe clinical and pathological data of 105 patients with advanced lung cancer and metastatic lung cancer treated by ¹²⁵I radioactive seed implantation in the Department of Thoracic Surgery, First Affiliated Hospital of Zhejiang University School of Medicine from January 1, 2014 to November 30, 2018. The patients were followed until March, 2019. The clinical efficacy and complications of seedimplantation were analyzed. RESULTS: A total of 105 patients were included in the study, 78 patients with advanced lung cancer and 27 patients with lung metastases. The median survival time after seed therapy was 395 days. The 1-year survival rate was about 78.1%, and the 2-year survival rate was about 56.1%. Seed implantation for advanced lung cancer is equivalent to lung metastasis. Seed combined with radiofrequency ablation, microwave ablation, and chemotherapy did not improve seed treatment. However, particle combined with external radiation therapy has a significant survival disadvantage compared with simple seed therapy. CONCLUSIONS The CT-guided ¹²⁵I radioactive seed implantation has controllable complications and can be used as a safe and effective treatment for advanced lung cancer and lung metastases.