Improving the Accuracy and Efficiency of Abnormal Cervical Squamous Cell Detection With Cytologist-in-the-Loop Artificial Intelligence.

Population-based cervical cytology screening techniques are demanding and laborious and have relatively poor diagnostic accuracy. In this study, we present a cytologist-in-the-loop artificial intelligence (CITL-AI) system to improve the accuracy and efficiency of abnormal cervical squamous cell detection in cervical cancer screening. The artificial intelligence (AI) system was developed using 8000 digitalized whole slide images, including 5713 negative and 2287 positive cases. External validation was performed using an independent, multicenter, real-world data set of 3514 women, who were screened for cervical cancer between 2021 and 2022. Each slide was assessed using the AI system, which generated risk scores. These scores were then used to optimize the triaging of true negative cases. The remaining slides were interpreted by cytologists who had varying degrees of experience and were categorized as either junior or senior specialists. Stand-alone AI had a sensitivity of 89.4% and a specificity of 66.4%. These data points were used to establish the lowest AI-based risk score (ie, 0.35) to optimize the triage configuration. A total of 1319 slides were triaged without missing any abnormal squamous cases. This also reduced the cytology workload by 37.5%. Reader analysis found CITL-AI had superior sensitivity and specificity compared with junior cytologists (81.6% vs 53.1% and 78.9% vs 66.2%, respectively; both with P < .001). For senior cytologists, CITL-AI specificity increased slightly from 89.9% to 91.5% (P = .029); however, sensitivity did not significantly increase (P = .450). Therefore, CITL-AI could reduce cytologists' workload by more than one-third while simultaneously improving diagnostic accuracy, especially compared with less experienced cytologists. This approach could improve the accuracy and efficiency of abnormal cervical squamous cell detection in cervical cancer screening programs worldwide.

Inhibition of invasion and metastasis in choriocarcinoma by migration and invasion inhibitory protein.

INTRODUCTION: Choriocarcinoma is a highly invasive gynaecologic malignancy. Molecular mechanism of metastasis in choriocarcinoma is poorly understood. Migration and invasion inhibitory protein (MIIP) regulates cell migration and invasion. Therefore, we aimed to elucidate the function of MIIP in choriocarcinoma. METHODS: Choriocarcinoma cell lines, JAR and JEG-3, were transfected with lentivirus carrying the MIIP-interfering RNA (to downregulate MIIP expression) or left untransfected (negative control). Cell migration and invasion were studied using transwell migration assays and scratch assays. In vivo tumour burden was studied using tumour xenograft models in specific-pathogen-free nude mice and live imaging. We elucidated possible molecular signalling pathways using western blotting. RESULTS: In transwell migration and scratch assays MIIP-downregulated JAR and JEG-3 cells migrated and invaded faster compared to their respective negative control cells. Migration and invasion by the MIIP-upregulated SWAN cells was slower than that by negative control SWAN cells. Live imaging revealed that bioluminescence values were higher in MIIP-downregulated tumours than in the negative control tumours. Mice with MIIP-downregulated tumours had higher serum human chorionic gonadotropin (HCG) levels than those with negative control tumours. The MIIP expression was negatively correlated with that of histone deacetylase (HDAC6) and positively correlated with that of acetylated α-tubulin. DISCUSSION: Thus, MIIP-by inhibiting cellular motility in choriocarcinoma-acts as a tumour suppressor gene. This highlights a potential therapeutic target for refractory choriocarcinoma. Additionally, HDAC6 and acetylated α-tubulin may be involved in the regulatory effects of MIIP on the biobehaviour of choriocarcinoma cells.

Evaluation and simplification of risk factors in FIGO 2000 scoring system for gestational trophoblastic neoplasia: a 19-year retrospective analysis.

OBJECTIVES: The International Federation of Gynecology and Obstetrics (FIGO) 2000 scoring system classifies gestational trophoblastic neoplasia (GTN) patients into low- and high-risk groups, so that single- or multi-agent chemotherapy can be administered accordingly. However, a number of FIGO-defined low-risk patients still exhibit resistance to single-agent regimens, and the risk factors currently adopted in the FIGO scoring system possess inequable values for predicting single-agent chemoresistance. The purpose of this study is therefore to evaluate the efficacy of risk factors in predicting single-agent chemoresistance and explore the feasibility of simplifying the FIGO 2000 scoring system for GTN. METHODS: The clinical data of 578 GTN patients who received chemotherapy between January 2000 and December 2018 were retrospectively reviewed. Univariate and multivariate logistic regression analyses were carried out to identify risk factors associated with single-agent chemoresistance in low-risk GTN patients. Then, simplified models were built and compared with the original FIGO 2000 scoring system. RESULTS: Among the eight FIGO risk factors, the univariate and multivariate analyses identified that pretreatment serum human chorionic gonadotropin (hCG) level and interval from antecedent pregnancy were consistently independent predictors for both first-line and subsequent single-agent chemoresistance. The simplified model with two independent factors showed a better performance in predicting single-agent chemoresistance than the model with the other four non-independent factors. However, the addition of other co-factors did improve the efficiency. Overall, simplified models can achieve favorable performance, but the original FIGO 2000 prognostic system still features the highest discrimination. CONCLUSIONS: Pretreatment serum hCG level and interval from antecedent pregnancy were independent predictors for both first-line and subsequent single-agent chemoresistance, and they had greater weight than other non-independent factors in predicting single-agent chemoresistance. The simplified model composed of certain selected factors is a promising alternative to the original FIGO 2000 prognostic system, and it shows comparable performance.

Lectin galactoside-binding soluble 3 binding protein mediates methotrexate resistance in choriocarcinoma cell lines.

Choriocarcinoma is one of the most aggressive gestational trophoblastic neoplasias (GTN). Methotrexate (MTX) resistance is the main cause of treatment failure in choriocarcinoma. However, the mechanism of MTX resistance in choriocarcinoma is poorly known. This study aims to explore the function of Lectin galactoside-binding soluble 3 binding protein (LGALS3BP) in MTX-resistance in choriocarcinoma cells. Gradual dose escalation of MTX was used to establish MTX-resistant choriocarcinoma cells (JAR-MTX and JEG3-MTX cell lines). RNA-sequencing was used to explore the differentially expressed genes. Plasmids or SiRNA transfection was used to regulate the expression of LGALS3BP. ELISA was used to detect the concentrations of LGALS3BP in the serum of MTX-sensitive and MTX-resistant patients. qRT-PCR, Western blot, and CCK-8 assay were used to determine the effects of LGALS3BP on MTX-resistance in JAR and JEG3 cells. The results showed the relative resistance index (RI) of MTX is 791.50 and 1040.04 in JAR-MTX and JEG3-MTX, respectively. LGALS3BP was up-regulated in MTX-resistant cells compared to original cells in both RNA and protein level. The concentrations of LGALS3BP were higher in the sera of MTX-resistant patients than in MTX-sensitive patients. Knocking down LGALS3BP can reverse the MTX-resistance in JAR-MTX and JEG3-MTX cells. In summary, we preliminarily established two MTX-resistant cells, and performed RNA-sequencing, and found LGALS3BP may play important role in MTX-resistance. Our work not only provides a research tool (MTX-resistant cells) for other researchers, but gives some hint on how MTX resistance is regulated.

Intramuscular Near-Infrared Spectroscopy for Muscle Flap Monitoring in a Porcine Model.

BACKGROUND: Current near-infrared spectroscopy (NIRS)-based systems for continuous flap monitoring are limited to flaps which carry a cutaneous paddle. As such, this useful and reliable technology has not previously been applicable to muscle-only free flaps where other modalities with substantial limitations continue to be utilized. METHODS: We present the first NIRS probe which allows continuous monitoring of local tissue oxygen saturation (StO2) directly within the substance of muscle tissue. This probe is flexible, subcentimeter in scale, waterproof, biocompatible, and is fitted with resorbable barbs which facilitate temporary autostabilization followed by easy atraumatic removal. This novel device was compared with a ViOptix T.Ox monitor in a porcine rectus abdominus myocutaneous flap model of arterial and venous occlusions. During these experiments, the T.Ox device was affixed to the skin paddle, while the novel probe was within the muscle component of the same flap. RESULTS: The intramuscular NIRS device and skin-mounted ViOptix T.Ox devices produced very similar StO2 tracings throughout the vascular clamping events, with obvious and parallel changes occurring upon vascular clamping and release. The normalized cross-correlation at zero lag describing correspondence between the novel intramuscular NIRS and T.Ox devices was >0.99. CONCLUSION: This novel intramuscular NIRS probe offers continuous monitoring of oxygen saturation within muscle flaps. This experiment demonstrates the potential suitability of this intramuscular NIRS probe for the task of muscle-only free flap monitoring, where NIRS has not previously been applicable. Testing in the clinical environment is necessary to assess durability and reliability.

MicroRNA-194 inhibits PRC1 activation of the Wnt/ß-catenin signaling pathway to prevent tumorigenesis by elevating self-renewal of non-side population cells and side population cells in esophageal cancer stem cells.

Esophageal cancer (EC) is a leading cause of cancer-related deaths worldwide. Recent studies highlight roles for microRNAs (miRNAs) in EC. Microarray analysis identified miR-194 as downregulated in EC. However, little is known about the role of miR-194 in regulating self-renewal or other biological properties of EC stem cells. RT-qPCR and Western blot confirmed the downregulation of miR-194 in EC stem cells and revealed the upregulation of protein regulator of cytokinesis 1 (PRC1) in EC. Dual-luciferase reporter assay confirmed miR-194 targeting of PRC1 resulting in its downregulation. MiR-194 overexpression or PRC1 silencing reduced PRC1 expression, preventing the activation of the Wnt/ß-catenin signaling pathway. Inhibition of the Wnt/ß-catenin signaling pathway prevented the proliferation, invasion, and self-renewal of EC stem cells while promoting apoptosis. Furthermore, overexpressing miR-194 or silencing PRC1 in nude mice decreased the tumor formation ability of EC stem cells in vivo. Taken together, miR-194 prevents the progression of EC by downregulating PRC1 and inactivating the Wnt/ß-catenin signaling pathway.

Endoscopic Esophageal Submucosal Tunnel Dissection for Cystic Lesions Originating from the Muscularis Propria of the Gastric Cardia.

OBJECTIVE: To analyze the types and properties of cystic lesions originating from the muscularis propria of the gastric cardia (CLMPGC), explore the growth pattern and anatomical characteristics, and evaluate the safety, feasibility, and clinical efficacy of endoscopic esophageal submucosal tunnel dissection (ESTD). METHODS: From September 2013 to July 2018, we treated 6 patients with CLMPGC whom we had diagnosed using endoscopy, endoscopic ultrasound (EUS), and Computed Tomography (CT) before the operations. ESTD was the best option for treatment for all these patients. Postoperative observation and follow-ups were performed, and the operational, clinical data, and treatment results are analyzed. RESULTS: The mean age of the patients was 50.67 ± 11.59 years (male : female = 1 : 1). The only clinical manifestations the patients exhibited were upper abdominal discomfort. The diameter of the masses was 2.05 ± 0.73 (1.1-3.0) cm. The duration of the ESTD operation was 93.5 (82-256) mins, and the length of hospital stay was 7.50 ± 1.38 days. Postoperative pathology showed 4 cases of an epithelioid cyst, and 2 cases of mucocele with xanthogranuloma. There were no complications, such as hemorrhage, pneumothorax, and pleural effusion during and after the operation. No recurrence during the follow-ups was observed. CONCLUSION: The CLMPGC were mainly mucocele and epidermoid cyst, in an expansive growth pattern, and these lesions had no distinct borders with the muscularis propria. The muscularis propria formed a complete wall of the lesion. There was no direct blood supply to the lesions from big blood vessels. Endoscopic esophageal submucosal tunnel dissection was a safe, feasible, and effective treatment for CLMPGC.

Endoscopic Submucosal Dissection of the Angiolipoma at Hypopharynx-Esophageal Introitus.

Angiolipoma in the region of the hypopharynx-esophageal introitus is a rare occurrence. Surgical treatment was performed in the few cases reported in the literature. Endoscopic submucosal dissection (ESD) is a minimally invasive treatment for hypopharyngeal and esophageal lesions. Our objective was to evaluate the feasibility, safety, and efficacy of ESD for treatment of angiolipoma at the hypopharynx-esophageal introitus. The patients with submucosal tumors at the hypopharynx-esophageal introitus were diagnosed as angiolipoma by preoperative evaluation with endoscopy, endoscopic ultrasonography, and computed tomography (CT). The patients who were diagnosed with angiolipoma agreed to undergo endoscopic submucosal dissection. Under general anesthesia and endotracheal intubation, ESD was used to remove the lesions. Preoperative, intraoperative, and postoperative data were collected and analyzed to evaluate the feasibility, safety, and effectiveness of endoscopic submucosal dissection. From January 2013 to December 2018, 6 cases of angiolipoma were treated with ESD with a success rate of 100%. The average operation time was 107.0 ± 69.4 minutes. Intraoperative blood loss is the main risk. Endoscopic thermocoagulation successfully stopped bleeding in all cases. Pharyngeal pain and painful swallowing were the main clinical signs. There was no stricture at the hypopharynx-esophageal introitus after the operation. ESD treatment of angiolipoma at hypopharynx-esophageal introitus is feasible, safe, and effective.

Knockdown of ST7-AS1 inhibits migration, invasion, cell cycle progression and induces apoptosis of gastric cancer.

Role of ST7-AS1 in the malignant progression of gastric cancer (GC) and its molecular mechanisms were investigated. ST7-AS1 level in GC tissues and matched normal tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Its level in GC patients presenting different tumor stages and tumor sizes was determined. Subsequently, ST7-AS1 level in epithelial cells of gastric mucosa and GC cell lines was examined. Cellular behavior of GC cells, including viability, apoptosis, migration, invasion and cell cycle, influenced by ST7-AS1 was evaluated. The interaction between ST7-AS1 and EZH2 was assessed by RNA immunoprecipitation (RIP) assay. The involvement of EZH2 in the progression of GC mediated by ST7-AS1 was identified. ST7-AS1 was upregulated in GC tissues and cell lines. Its level was positively correlated to tumor stage and tumor size of GC. Knockdown of ST7-AS1 attenuated proliferative, migratory and invasive abilities, arrested cell cycle progression and induced apoptosis of GC cells. EZH2 was identified to interact with ST7-AS1, which attenuated the regulatory effects of ST7-AS1 on migratory and invasive abilities of GC cells. Upregulated ST7-AS1 in GC accelerated proliferation, migration and invasion, and inhibited apoptosis, thus aggravating the progression of GC.

shRNA-induced silencing of Ras-related C3 botulinum toxin substrate 1 inhibits the proliferation of colon cancer cells through upregulation of BAD and downregulation of cyclin D1.

Ras-related C3 botulinum toxin substrate 1 (RAC1) is a member of the Rho family of small GTPases. Recent studies have reported that RAC1 serves an important role in colon cancer cell proliferation. The present study aimed to investigate the effects of RAC1 knockdown on cell proliferation, cell cycle progression and apoptosis of colon cancer cells. Lentivirus­mediated short hairpin RNA (shRNA) was used to knockdown RAC1 expression in colon cancer cell lines, and cell proliferation, apoptosis, cell cycle progression were evaluated by MTT assays and flow cytometry. The differences in mRNAs expression were identified between RAC1-knockdown cells and control cells using a mRNA microarray, following which quantitative PCR (qPCR) and western blot were employed to confirm the results of the mRNA microarray. The proliferative ability of colon cancer cells was significantly decreased following RAC1 knockdown, and RAC1 knockdown increased the apoptotic rate and enhanced cell cycle arrest at G1 phase in colon cancer cells. In addition, >1,200 known genes were demonstrated to be involved in RAC1­associated tumorigenic functions in SW620 colon cancer cells, as determined by gene chip analysis; these genes were associated with cell proliferation, cell cycle, apoptosis and metastasis. Furthermore, western blot analysis indicated that cyclin D1 was downregulated, whereas B­cell lymphoma 2­associated agonist of cell death (BAD) was upregulated following RAC1 knockdown in colon cancer cells. In conclusion, RAC1 silencing may suppress the proliferation of colon cancer cells by inducing apoptosis and cell cycle arrest. In addition, a large number of genes were revealed to be involved in the process, including BAD, which was upregulated and cyclin D1, which was downregulated. Further studies on these differentially expressed genes may provide a better understanding of the potential roles of RAC1 in colon carcinogenesis.

Expression profile of circular RNAs in human gastric cancer tissues.

Circular RNAs (circRNAs) represent a newly identified class of non­coding RNA molecules, which interfere with gene transcription by adsorbing microRNAs (miRNAs). CircRNAs serve important roles in disease development and have the potential to serve as a novel class of biomarkers for clinical diagnosis. However, the role of circRNAs in the occurrence and development of gastric cancer (GC) remains unclear. In the present study, the expression profiles of circRNAs were compared between GC and adjacent normal tissues using a circRNA microarray, following which quantitative polymerase chain reaction (qPCR) was used to confirm the results of the circRNA microarray. Compared with the adjacent, normal mucosal tissues, 16 circRNAs were upregulated and 84 circRNAs were downregulated in GC. A total of 10 circRNAs were selected for validation in three pairs of GC and adjacent noncancerous tissues. The qPCR results were consistent with the findings of the microarray­based expression analysis. Of the circRNAs studied, only circRNA­0026 (hsa_circ_0000026) exhibited significantly different expression in GC (2.8­fold, P=0.001). Furthermore, online Database for Annotation, Visualization and Integrated Discovery annotation was used to predict circRNA­targeted miRNA­gene interactions. The analysis revealed that circRNA­0026 may regulate RNA transcription, RNA metabolism, gene expression, gene silencing and other biological functions in GC. In conclusion, differential expression of circRNAs may be associated with GC tumorigenesis, and circRNA­0026 is a promising biomarker for GC diagnosis and targeted therapy.

Increased NOD1, but not NOD2, activity in subcutaneous adipose tissue from patients with metabolic syndrome.

OBJECTIVE: Nucleotide-binding oligomerization domain (NOD) protein, as cytoplasmic receptor of the innate immune response, plays an important role in adipose inflammation and insulin resistance in obesity. Our objective was to examine adipose tissue (AT) NOD in nascent metabolic syndrome (MetS) patients and to investigate its association with MetS features. METHODS: Thirty-four MetS subjects and 31 controls were recruited. Fasting blood was collected, and abdominal subcutaneous AT was obtained by biopsy for NOD1/NOD2 expression and activity. RESULTS: MetS subjects showed significantly increased expression for NOD1 on adipose depots as compared to controls. In addition to increased expression of downstream signaling mediators RIPK2 and NF-κB p65 nuclear translocation, there was remarkably higher release of monocyte chemotactic protein1 (MCP-1), interleukin (IL)-6, and IL-8 in MetS versus controls following priming of the isolated adipocytes with NOD1 ligand iE-DAP. With regard to NOD2, the differences between the two groups were not significant in either basal state or after activation. Increased NOD1 positively correlated with waist circumference. NOD1 was also correlated with HbA1c and HOMA-IR. NOD1 positively correlated with serum levels of IL-6, MCP-1, and NF-κB activity. CONCLUSIONS: Activation of the innate immune pathway via NOD1 may be partially responsible for the increased systemic inflammation and insulin resistance in MetS.

Monitoring thermal-induced changes in tumor blood flow and microvessels with laser speckle contrast imaging.

Laser speckle contrast imaging (LSCI) was used to monitor thermal-induced changes in the blood flow and the diameter of mesenteric microvessels of normal and tumor bearing mice under 60 min treatment at different constant temperatures between 41 degrees C and 45 degrees C. The results show that the blood flow and the diameter increase at the beginning and then reach a plateau and finally start to decrease. The lower the temperature, the longer the plateau stays. A t-test indicates that there is no significant difference in plateau values of relative blood flow and relative diameter for the same group. For normal mice, the relative increases in the blood flow and the diameter are 1.26 and 1.41, respectively, while for tumor-bearing mice they are 1.08 and 1.13, respectively. At higher treatment temperature or under longer heat treatment, there are decreases in the blood flow and the diameter, such changes in tumor-bearing mice are more obvious than those in normal mice, which means tumor microvessels are more sensitive to heat than normal. Moreover, thermal induced shrink of microvessel usually occurs sooner than the decrease in blood flow, and the relative change in diameter is larger than that in blood flow. Therefore we may conclude that deformation of vessel is a main factor for changing the blood perfusion of a microvessel.

[Factors affecting the prognosis of diffuse large B-cell lymphoma in Chinese].

OBJECTIVE: To study the correlation between clinical prognosis and clinicopathologic features, origin and cell proliferous index of diffuse large B-cell lymphoma (DLBCL) in China. METHODS: The data of 60 cases of DLBCL were collected with the study of international prognostic index (IPI) and the follow-up results. Immunohistochemistry stain was used to check the expression of CD(45)RO, CD(3), CD(20), CD(79)a, CD(45)RA, Ki-67, p53 and bcl-6. RESULTS: The ratio of male to female is 1.73 and the average age is 53.1. In 53.3% cases lymph node were involved (32/60), gastric intestinal tract and tonsil were the most common extranodal sites of involvement. 23 patients 38.3% died during follow-up, the longest survival period lasting 108 months.16 died in the first year after establishment of diagnosis (16/23, 69.6%). IPI is an independent prognostic index; the lower the index, the better the prognosis (P = 0.0102). Positive incidence of CD(20), CD(79)a and CD(45)RA was 91.5%, 73.7% and 58.3%. Ki-67, p53, bcl-6 were expressed in some cases (48/53, 90.6%; 26/46, 56.5%; 21/41, 51.2%). The expression of bcl-6 protein was somewhat related with prognosis (P = 0.0049), but the expression of p53 was not (P = 0.5948). CONCLUSIONS: The clinicopathologic features of DLBCL are similar in the East and the West. It is highly aggressive tumor. Most of the cases died in the first year after establishment of diagnosis. IPI can be used to predict the clinical outcome. The expression of bcl-6 protein was somewhat related with clinical prognosis, but that of p53 was not.

[Expression of nm23 and proliferating cell nuclear antigen (PCNA) in human brain gliomas and their significance].

BACKGROUND & OBJECTIVE: Brain gliomas seldom undergo extracranial metastasis. Local recurrence is the main reason of tumor patient's death. Therefore, it is important to detect tumor biological features through determination of gene expression. This study was designed to investigate the expression of nm23 (non-metastasis gene, nm23)and PCNA (proliferating cell nuclear antigen) and evaluate the malignancy, recurrence, and prognosis of the tumor. METHODS: In 50 specimens of different malignant gliomas,the expression of nm23 and PCNA were examined using SP immunohistochemical staining. RESULTS: (1)The label indexes of nm23 and PCNA in low-grade gliomas were 3.40+/-0.27 and 3.60+/-0.05, respectively; while the label indexes of nm23 and PCNA in high-grade gliomas were 1.72+/-0.18 and 6.20+/-0.23, respectively.There was significant difference between the two groups(P< 0.05). (2)The positive rates of nm23 and PCNA were 56% (14 cases) and 64% (16 cases) in 25 cases of low-grade gliomas, while the positive rates of nm23 and PCNA were 12% (3 cases) and 88% (22 cases) in 25 cases of high-grade gliomas. There was significant difference between the two groups (P< 0.05). (3)The positive rates of nm23 and PCNA were 0% (0 cases) and 100% (9 cases) in 9 cases of recurrent gliomas, while the positive rates of nm23 and PCNA were 50%(34 cases) and 50%(4 cases) in 8 cases of non-recurrent gliomas. There was significant difference between the two groups (P< 0.05). (4)The label indexes of nm23 and PCNA in gliomas were inversely correlated (r=-0.5335,P< 0.001). CONCLUSION: (1)The expression of nm23 is inversely correlated with the malignancy of gliomas,i.e.the lower expression indicates the higher malignancy. (2)The expression of PCNA is associated with the increased malignancy. (3)Both nm23 and PCNA may be useful biological markers to evaluate the malignancy and prognosis of patients with gliomas.