A one-stage deep learning based method for automatic analysis of droplet-based digital PCR images.

Droplet-based dPCR offers many advantages over chip-based dPCR, such as lower processing cost, higher droplet density, higher throughput, while requiring less sample. However, the stochastic nature of droplet locations, uneven illuminations, and unclear droplet boundaries make automatic image analysis challenging. Most methods currently used to count a large amount of microdroplets rely on flow detection. Conventional machine vision algorithms cannot extract all information of the targets from complex backgrounds. Some two-stage methods, which first locate and then classify droplets according to their grayscale values, require high-quality imaging. In this study, we addressed these limitations by improving a one-stage deep learning algorithm named YOLOv5 and applying it to the detection task to realize one-stage detection. We introduced an attention mechanism module to increase the detection rate of small targets and used a new loss function to speed up the training process. Furthermore, we employed a network pruning method to facilitate the deployment of the model on mobile devices while preserving its performance. We validated the model with captured droplet-based dPCR images and found that the improved model accurately identified negative and positive droplets in complex backgrounds with an error rate of 0.65%. This method is characterized by its fast detection speed, high accuracy, and ability to be used on mobile devices or cloud platforms. Overall, the study presents a novel approach for detecting droplets in large-scale microdroplet images and provides a promising solution for accurate and efficient droplet counting in droplet-based dPCR.

vNOTES Hysterectomy with Sentinel Lymph Node Mapping for Endometrial Cancer: Description of Technique and Perioperative Outcomes.

STUDY OBJECTIVE: To explore the technique and clinical value of transvaginal natural orifice transluminal endoscopic surgery (vNOTES) in hysterectomy and sentinel lymph node (SLN) mapping for endometrial cancer by comparing its perioperative outcomes with those of laparoscopic staging. DESIGN: Retrospective cohort study. SETTING: Department of gynecology at a tertiary medical center. PATIENTS: All women diagnosed with endometrial cancer who underwent minimally invasive surgery at our center between August 2017 and May 2020. INTERVENTIONS: Both vNOTES and laparoscopic approaches were used for hysterectomy and SLN mapping. The success of SLN detection as well as perioperative outcomes were subsequently analyzed. MEASUREMENTS AND MAIN RESULTS: This study included 74 patients; 23 patients underwent vNOTES surgery, whereas 51 underwent standard laparoscopic surgery. The total successful SLN detection was 95.7% in the vNOTES group and 92.2% in the laparoscopy group (p >.05), whereas the bilateral success rates were 87.0% and 90.2%, respectively. No difference in SLN detection was observed between the 2 groups in terms of the side-specific mapping efficacy quotient (91.3% vs 91.2%, p = .47). The number of harvested SLNs, operative time, estimated blood loss, and intraoperative complications in the 2 groups were similar. One (4.3%) postoperative complication occurred in the vNOTES group vs 4 (7.9%) in the laparoscopy group (p = .029), and the median postoperative hospital stay was 3 days vs 4 days (p = .003). CONCLUSION: This study suggests that the vNOTES procedure is feasible, with a potentially decreased postoperative hospital stay, faster recovery, and better cosmetic results. However, prospective research is needed to validate its broader clinical application.

[Pathologic features of fallopian tubal fimbriae in patients with endometrial serous carcinoma].

OBJECTIVE: To discuss the potential relationship between endometrial serous carcinoma (ESC) and tubal epithelial lesions by pathologic examination of fallopian tubes with ESC. METHODS: A total of 30 cases of typical ESC were reexamined and chosen by the pathologist. In each case, bilateral fallopian tubes were submitted to examination of pathologic morphology and immunostaining for p53, annexin IV (ANX-IV), human epidermal growth factor receptor 2 (HER2)/neu, and high-mobility group protein A2 (HMGA2). RESULTS: Fallopian tubal epithelial lesions were found in 15 cases, including 9 cases tubal serous carcinoma, 2 cases serous tubal intraepithelial carcinoma (STIC) and 2 cases epithelial hyperplasia. Both sides of tubal serous carcinoma and STIC were found in 1 case. The results showed the positive expression for p53 in 26 (87%) out of 30 endometrial malignant specimens tissues and 9 (30%) tubal tissues samples (P > 0.05). Twenty-five (83%) endometrial malignant specimens tissues and 6 (20%) tubal tissues samples showed the positive expression of ANX-IV. Twenty-one (70%) endometrial malignant tissues and 7 (23%) tubal tissues showed the positive expression of HER2/neu. Twenty-five (83%) endometrial malignant tissues and 6 (20%) tubal tissues showed the positive expression of HMGA2. While, there were significant differences among the expression of three proteins between endometrium and the fallopian tube site (all P < 0.05). CONCLUSIONS: STIC may be associated with the occurrence of ESC. The expression of p53 was positively correlated between the fallopian tube and the endometrium. ANX-IV, HER2/neu and HMGA2 were extensively expressed in ESC.

[Relationship between the expression of dual specificity phosphatase-1 and the prognosis of endometrioid adenocarcinoma].

OBJECTIVE: To clarify the relationship between the expression of dual specificity phosphatase-1 (DUSP1) and the prognosis of endometrioid adenocarcinoma. METHODS: The expression of DUSP1 was determined by immunohistochemical staining in specimens from 81 patients with endometrial carcinoma undergoing surgical resection. The relationship between DUSP1 expression, clinicopathological factors and prognosis were further evaluated. RESULTS: In 81 endometrioid carcinoma samples, 59 (72.84%) cases were positive while 22 negative.Except for lymph node metastasis, the expression of DUSP1 was correlated with FIGO stage, tumor grade, myometrial invasion and the expressions of estrogen receptor (ER) and progesterone receptor (PR) (P < 0.05) .Kaplan-Meier analysis showed that patients with a positive DUSP1 expression had significantly prolonged 5-year disease-free survival rates of 98.2% and 76.0% respectively (P < 0.05). And COX regression analysis revealed that the expressions of DUSP1 and PR were independent prognostic indicators of endometrioid carcinoma, the HR (hazard ratio) of DUSP1 negative expression was 21.2.Spearman analysis further showed that the expression of DUSP1 was positively correlated with PR (r = 0.256, P < 0.05). CONCLUSION: DUSP1 may be a potential negative prognostic indicator for endometrioid adenocarcinoma.

[Significance of prognostic evaluation of International Federation of Gynecology and Obstetrics 2009 staging system on stage I endometrioid adenocarcinoma].

OBJECTIVE: To explore the impact of 2009 International Federation of Gynecology and Obstetrics (FIGO) staging system alteration for stage I endometrioid adenocarcinoma on its' prognosis assessing. METHODS: A retrospective study was carried out on 244 cases with endometrial carcinoma admitted in Peking University People's Hospital from Jan.1995 to Feb.2008. RESULTS: (1) All 244 patients were divided into FIGO 2009 Ia group (n = 200) and FIGO 2009 Ib group (n = 44) according to FIGO 2009 staging system, while they were divided into FIGO 1988 Ia group (n = 34), FIGO 1988 Ib group (n = 156) and FIGO 1988 Ic group (n = 29). The others 25 cases were stage IIa (n = 16) and stage IIIa with merely positive abdominal cytology (n = 9) according to FIGO 1988 staging system.(2) The higher percentage of low-grade in FIGO 1988 Ia group than that in FIGO 2009 Ia group (P = 0.003). Compared with FIGO 2009 Ia group, the age of the patients, surgery extent, the percentage of lymph node excision and received chemotherapy and radiotherapy, there were no difference in FIGO 1988 Ia and Ib group, respectively (P > 0.05). There were 5.9% (2/34) and 6.7% (10/150) found relapse among FIGO 1988 Ia group and FIGO 1988 Ib group, and there were 2.9% (1/34) and 2.7% (4/150) for the two groups died of carcinoma. Compared with FIGO 2009 Ia group, there were not significant difference [7.5% (13/200) vs. 3.0% (6/200); P > 0.05]. The 5 years and 10 years progression-free survival (PFS) of FIGO 1988 Ia group and Ib group were (97.0 ± 3.0)%, (90.9 ± 6.5)% and (95.3 ± 2.1)%, (90.2 ± 3.6)%, respectively, in which there were not significant difference compared with that in FIGO 2009 Ia group [(96.1 ± 1.6)%, (89.6 ± 3.2)%; P > 0.05]. The 5 years and 10 years overall survival (OS) in FIGO 1988 Ia group and Ib group were 100%, (93.8 ± 6.0)% and (96.9 ± 1.8)%, (95.2 ± 2.5)%, respectively, in which there were did not significant difference with that in FIGO 2009 Ia group [(97.9 ± 1.2)%, (93.4 ± 2.8)%; P > 0.05].(3) There were not significant difference between FIGO 1988 Ic group and FIGO 2009 Ib group (P > 0.05) for the age of the patients, grade, surgery extent, lymph node excision, the percentage of received chemotherapy and radiotherapy. Between FIGO 1988 Ic group and FIGO 2009 Ib group, there were 3.4% (1/29) and 6.8% (3/44) cases found relapse, respectively. And there were 0 and 2.3% (1/44) cases died of carcinoma in the two groups, in which there were not differ much either (P > 0.05). The 5 years and 10 years PFS in FIGO 1988 Ic group were all 100%, while they were 100% and (90.9 ± 6.2)% in FIGO 2009 Ib group. The 5 years and 10 years OS in FIGO 1988 Ic group were all 100%, but were 100% and (95.0 ± 4.9)% in FIGO 2009 Ib group, in which they all did not significantly differ much (P > 0.05). (4) The patients in FIGO 2009 Ia group were younger than those in FIGO 2009 Ib group (P < 0.01). The percentage of low grade in FIGO 2009 Ia group were higher than that in FIGO 2009 Ib group (P = 0.029). The percentages of received chemotherapy and radiotherapy in FIGO 2009 Ia group were lower than that in FIGO 2009 Ib group remarkably (P < 0.01). But there were not significant difference in the uterine excision extent and the percentage of lymph node excision between the two groups (P > 0.05). There were not significantly differ in the relapse rates and the death rates between the FIGO 2009 Ia group and FIGO 2009 Ib group (P > 0.05). There were also not significant difference in PFS and OS between the two groups (P > 0.05). CONCLUSIONS: There were not significant difference in the prognosis between FIGO 2009 stage Ia and FIGO 1988 stage Ia and Ib. There were also not significant difference in the prognosis between FIGO 2009 stage Ia and FIGO 2009 stage Ib, which may be due to received more chemotherapy and radiotherapy in FIGO 2009 stage Ib patients.

Chopstick technique versus cross technique in LESS hysterectomy (CCLEH study): a prospective randomized controlled trial.

BACKGROUND: The traditional cross technique can be used to complete most laparoendoscopic single-site surgery (LESS) procedures, but some relatively precise operations, such as vaginal stump suturing, are challenging. In practice, we have introduced a novel technique named the chopstick technique and applied it to more complex operations, such as cervical cancer operations, and found that it contributes to performing delicate surgery. The efficacy and safety of two different surgical techniques in LESS hysterectomy remain to be validated. METHODS: Patients who undergo total hysterectomy will be enrolled in this RCT. Stratified randomization will be performed according to uterine size (< 10 cm, 10-15cm, ≥ 15 cm). The participants will be divided into the chopstick technique group or cross technique group to undergo laparoendoscopic single-site total hysterectomy (LESS-TH), and then the perioperative and postoperative data, including the total operation time and other times, transfer rates, estimated blood loss, surgeon fatigue, intraoperative and postoperative complications (within 8 weeks after surgery), health-related quality of life (EQ-5D) scores, postoperative hospital stay, and hospitalization expenses, will be evaluated. The primary outcome is the operating time for total hysterectomy under LESS, and the other outcomes are secondary outcomes. DISCUSSION: It is expected that the efficacy of the two techniques in LESS, the chopstick technique vs. the cross technique, will be compared and accumulate safety data on the new techniques will be accumulated. TRIAL REGISTRATION: ChiCTR2000040843, registered on June 16,2020 Protocol version: Version 2.0; Date: 2020.05.10.

A preoperative scoring system to predict the probability of laparoendoscopic single-site extracorporeal cystectomy in patients with benign ovarian cysts.

Objective: To develop a preoperative scoring system (PSS) to predict whether laparoendoscopic single-site extracorporeal (LESS-E) cystectomy can be performed in patients with benign ovarian cysts. Method: We reviewed data on patients who underwent LESS cystectomy between August 2016 and October 2019 at the first Affiliated Hospital, Army Medical University. The independent predictors of LESS-E cystectomy in patients with benign ovarian cysts were identified using multivariate logistic regression analyses. A nomogram for predicting LESS-E cystectomy in patients with benign ovarian cysts was developed, and to simplify the score, we establish a preoperative scoring system to guide the choice of surgical approach in patients with highly probable benign ovarian cysts. Results: Our analysis showed that age, BMI, height and the diameter of ovarian cysts were independent predictors of LESS-E cystectomy. A nomogram was developed based on these four factors, which had a concordance index of 0.838 and R 2 = 0.415. To simplify the score, the predicted indicators in the regression model were scored by dividing the beta coefficient by the absolute value of the minimum beta coefficient, and the sum of each predictor score established a PSS. In the total set, the selected cutoff value according to the maximum point of the Youden index was 8, and a preoperative score ≥ 8 identified patients undergoing LESS-E cystectomy with a positive predictive value of 67.4% and a negative predictive value of 88.6%. Conclusion: A PSS to predict the chances of LESS-E cystectomy was established. This system could be helpful for selecting the appropriate surgical strategy for patients with benign ovarian cysts.

Learning curve for laparoendoscopic single-site radical hysterectomy using the "chopstick" technique: a retrospective cohort study.

Background: Laparoendoscopic single-site surgery (LESS), as a promising minimally invasive surgery, is confronted with the obstacle of the "chopstick effect" which limits its further application. The "chopstick" technique is characterized by the usage of instruments of parallel and equal length, and in operations relying on double-fulcrum and unique surgeon position can play a key role in overcoming the above disadvantage effect. This study sought to explore the learning curve for the use of the novel "chopstick" technique in laparoendoscopic single-site radical hysterectomy (LESS-RH) and evaluate the technique's practicability. Methods: Consecutive cervical cancer patients who underwent LESS-RH with the "chopstick" technique by a surgeon with rich experience in laparoscopy from November 2016 to September 2018 were included in the study. The learning curve of his surgeries with the "chopstick" technique was evaluated using the cumulative summation (CUSUM) method by analyzing operation time (OT) which was the surrogate indicator of surgical ability. The patients were divided into Phase I and Phase II based on the learning curve peak, whose demographic and perioperative characteristics, such as tumor Federation International of Gynecology and Obstetrics (FIGO) stage and histology, operation time, blood loss, and complications were collected and analyzed. Results: The mean OT was 231.5 min (range, 115-355 min). The division of the learning curve based on OT occurred after the first 15 cases were finished, dividing Phase I and Phase II. The mean OT for Phase I (259 min) was significantly longer than that of Phase II (219 min) (P=0.02). Only 1 intraoperative complication occurred in Phase I, and none occurred in Phase II. Major postoperative complications occurred more frequently in Phase I (N=3) than in Phase II (N=0). No significant differences were observed in terms of lymph nodes, blood loss, or pathological features. Conclusions: The "chopstick" technique may help surgeons obtain stable LESS surgical performance through a relatively short learning curve, even in some complex surgeries, such as radical hysterectomy.

LIFGO: A modular laser-induced fluorescence detection system based on plug-in blocks.

In this work, a laser-induced fluorescence (LIF) detection system built in a modular assembling mode was developed based on commercial LEGO blocks and 3D printed blocks. We designed and fabricated a variety of 3D printed building blocks fixed with optical components, including laser light source, filters, lens, dichroic mirror, photodiode detector, and control circuits. Utilizing the relatively high positioning precision of the plug-in blocks, a modular construction strategy was adopted using the flexible plug-in combination of the blocks to build a highly sensitive laser-induced fluorescence detection system, LIFGO. The LIFGO system has a simple structure which could be constructed by inexperienced users within 3 h. We optimized the structure and tested the performance of the LIFGO system, and its detection limits for sodium fluorescein solution in 100 µm i.d. and 250 µm i.d. capillaries were 7 nM and 0.9 nM, respectively. Based on the LIFGO system, we also built a simple capillary electrophoresis (CE) system and applied it to the analysis of DNA fragments to demonstrate its application possibility in biochemical analysis. The separation of 7 fragments in DL500 DNA markers were completed in 600 s. Because of the features of low cost (less than $100) and easy-to-build construction, we introduced the LIFGO system to the experimental teaching of instrumental analysis for undergraduate students. The modular construction form of the LIF detection system greatly reduces the threshold of instrument construction, which is conducive to the popularization of the LIF detection technique in routine laboratories as well as the reform of experimental teaching mode.

cDNA microarray analysis and immunohistochemistry reveal a distinct molecular phenotype in serous endometrial cancer compared to endometrioid endometrial cancer.

PURPOSE: The main objective of this study was to refine more precisely the gene expression patterns used to distinguish serous from endometrioid endometrial carcinoma. METHODS: A low-density cDNA microarray containing 492 genes was designed and constructed. The gene expression profiles of 32 endometrioid and 5 serous endometrial cancer tissue samples were compared. The expression of 5 differentially expressed genes: NDC80, BUB1, FUT8, ANXA4 and BBC3 in endometrioid and serous adenocarcinoma samples was further evaluated by quantitative real-time PCR and immunohistochemistry. RESULTS: Unsupervised cluster analysis revealed that the 5 serous adenocarcinomas clustered together. These were separated from the endometrioid adenocarcinomas which were further sorted into 3 additional clusters. A comparative analysis indicated that there was a significant difference in FIGO stage with no significant difference in depth of myometrial invasion among the 4 clusters. The FIGO ternary grading system could not distinctly separate the 3 clusters of endometrioid adenocarcinomas, but a binary grading system was able to do so. Using a supervised analysis, we have identified 46 genes exhibiting >2-fold differences that can be used to statistically differentiate serous adenocarcinomas from endometrioid adenocarcinomas. The directions of gene and protein expression change of five differentially expressed genes estimated by real-time PCR and immunohistochemistry are consistent with those estimated from microarray. CONCLUSIONS: Serous adenocarcinoma exhibits distinct gene expression profiles, compared with those of endometrioid adenocarcinoma. These differences make it feasible to validate microarray data by immunohistochemistry, and they will ultimately allow us to identify tumors according to their immunohistochemical phenotype. The accuracy of classifying endometrial tumors using a system based on their gene expression patterns is much higher than the accuracy of the FIGO grading system. Thus, this gene expression pattern-based system may prove to be crucial in developing novel treatment strategies for endometrial cancers at the molecular level in future.

[Relationships between the molecular biomarkers and the clinicopathologic features and prognosis in endometrial carcinoma].

OBJECTIVE: To explore the relationships between the expressions of estrogen receptor (ER), progestin receptor (PR), phosphatase and tension homology deleted on chromosome ten (PTEN), p53, Ki-67 and the clinicopathologic features and prognosis in endometrial carcinoma. METHODS: The data of clinical characteristics, pathological types, histological grades, follow-ups and the expressions of molecular markers detected by immunohistochemistry, and collected from 200 patients with primary endometrial carcinoma, were analyzed. RESULTS: (1) In the cases of endometrial carcinoma, the expression rates of ER, PR, PTEN, p53 were 86.5%, 85.5%, 82.1%, and 49.2%, respectively. The expression level of Ki-67 in the tumor tissues was 46.9% ± 24.7%. (2) A negative correlation was observed between the gravidity and the expression of PR (r=-0.191, P=0.007). On the other hand, age and parturition time were in positive correlation with the expression of p53 (r=0.184, P=0.041; r=0.255, P=0.004). (3) The expression rates of ER, PR and p53 in the endometrioid carcinoma exhibited significant differences comparing with other types (P<0.01). (4) A negative correlation was found between the expression of ER and the FIGO staging (r=-0.176, P=0.013). The positive rate of ER in the cases with Stage I was higher than that in cases with Stage II and above (P=0.015). (5) A negative correlation was found between the histological grade and the expressions of ER and PR (r=-0.217, P=0.002; r= -0.317, P=0.000), however, a positive correlation was detected between the grade with the expressions of p53 and Ki-67 (r=0.327, P=0.000; r=0.465, P=0.000). Compared with the grade 3 tumors, the other grades exhibited significant different expression levels of ER, PR, p53, and Ki-67 (P<0.01). (6) A negative correlation was observed between the depth of myometrial invasion and the positive rate of ER (r=-0.142, P=0.047). There were statistically significant different expression rates of ER and PR between the cases whether the cancer invaded the deep myometrium or not (P<0.05). (7) Multivariate survival analysis showed that patients with PR (+) had longer overall survival than those with PR (-) (P=0.011). CONCLUSION: The immunohistochemical study of endometrium samples obtained from dilatation and curettage of uterine will be beneficial to the understanding of the clinicopathologic features of the endometrial carcinoma before the operation. The value of estimating the prognosis using the expressions of ER, PTEN, p53 and Ki-67 was negative, except for the expression of PR.

[Clinical analysis on the lymph nodes metastasis characters and their relation with the prognosis of the endometrial carcinoma patients].

OBJECTIVE: To explore the lymph nodes (LN) metastasis characters of the endometrial carcinoma and its relation with the patients' prognosis. METHODS: A retrospective study was carried out on 227 cases of endometrial carcinoma who admitted to our department and underwent LN excision from Jul.2000 to Feb.2008. RESULTS: Among 227 cases who underwent pelvic LN excision, there were 22 cases (9.7%) presented LN metastasis. There were 12 cases with positive external iliac LN from 20 cases of patients with data in LN grouping. Para-aortic LN excision was carried out on 138 patients. There were 6 cases with positive para-aortic LN, 5 cases of them together with pelvic LN metastasis. Those patients with cervix involvement, annex metastasis, deep myometrium infiltration, grade 2-3 and negative estrogen receptor occurred pelvic LN metastasis more frequently than the others (P < 0.05). Among the 6 cases with positive para-aortic LN, there were 3 cases (3/6) with deep myometrium infiltration. For those whose para-aortic LN was negative, it was only 16.7% (22 cases). But there were no difference statistically between them (P > 0.05). There were significant difference in 3 years disease-free survival rate between patients with positive pelvic LN or negative pelvic LN [(81.8 ± 8.2)% vs (97.4 ± 1.2)%, P = 0.004]. While there were not significant difference in 3 years disease-free survival rate between patients with positive para-aortic LN or negative para-aortic LN [100% vs (96.7 ± 1.6)%, P > 0.05]. Single factor analysis showed that the age more than 50 years, annex metastasis and pelvic LN metastasis related with the recurrence (P < 0.01). But cervix involvement, deep myometrium infiltration, para-aortic LN metastasis, pathology type, tumor grade and estrogen receptor did not relate with the recurrence (P > 0.05). Cox regression analysis showed that annex metastasis and the age of patients were independent risk factors affecting the recurrence (P = 0.011, P = 0.025). CONCLUSIONS: The most common site of pelvic LN metastasis is the external iliac LN for endometrial carcinoma patients. The patients with positive para-aortic LN always accompanied pelvic LN metastasis. Those patients with cervical involvement, annex metastasis, deep myometrium infiltration, poor differentiation and negative estrogen receptor be more likely exist pelvic LN metastasis. Pelvic LN metastasis may affect the prognosis of endometrial carcinoma patients.

Chopstick technique used in laparoendoscopic single site radical hysterectomy for early stage cervical cancer.

Laparoendoscopic single-site surgery (LESS) further minimizes the invasiveness of traditional laparoscopic surgery. However, the "chopstick" effect caused by the parallel arrangement of the instruments in the umbilicus is considered an obstacle indelicate operations. The purpose of this study was to introduce a new technique characterized by a double fulcrum formed by instruments, named the "chopstick" technique, which facilitates the expedient accomplishment of complicated surgeries such as LESS radical hysterectomy (LESS-RH). Seventy-three patients who underwent LESS-RH using the "chopstick" technique were retrospectively analyzed. The procedure was performed successfully in 72 patients. The median operative duration was 225 min, and the median intraoperative blood loss was 200 ml. Among the operations in the first 20 patients, intraoperative vascular injuries and bladder injury occurred in two patients and were repaired by LESS. Patients responded positively regarding minimal postoperative pain control. The score of satisfaction with the cosmetic outcome expressed by the patients was eight at discharge and nine 30 days later. In conclusion, this study presents the feasibility of accomplishing complicated procedures, such as radical hysterectomy, by LESS using the "chopstick" technique. This approach provides more options for both selected patients and surgeons.

Perioperative outcomes of laparoendoscopic single-site extracorporeal versus intracorporeal cystectomy for benign ovarian cysts: a retrospective cohort study based on propensity score matching.

BACKGROUND: Ovarian cysts are the most common gynecological disease, and laparo-endoscopic single-site (LESS) cystectomy is a popular surgical procedure. A new laparoscopic technique combining the advantages of LESS surgery and open surgery has been introduced to treat large ovarian cysts. To our knowledge, no previous research has compared LESS-extracorporeal (LESS-E) cystectomy to LESS-intracorporeal (LESS-I) cystectomy. This study compared the perioperative results of LESS-E cystectomy and LESS-I cystectomy in the treatment of benign ovarian cysts. METHODS: Two hundred eighty-eight cases of cystectomy from our institutional database were retrospectively reviewed. 1:1 propensity score matching (PSM) was performed to minimize bias due to any imbalanced baseline features between the 2 groups, which were matched in terms of age, body mass index, and the largest diameter of ovarian cysts. Seventy-nine cases were then selected from each group, and the perioperative outcomes of the 2 cohorts were analyzed. RESULTS: The mean (standard deviation) largest diameter of ovarian cysts was 8.30 (3.56) cm in the LESS-I group and 9.14 (3.15) cm in the LESS-E group (P=0.118). No statistically significant difference was found between the 2 groups in terms of estimated blood loss, postoperative hemoglobin decline, postoperative pain in 24 hours, postoperative hospital stay, and total hospital costs (P>0.05). However, the mean operation time of the LESS-E group was shorter than that of the LESS-I group (85.01 vs. 104.25 minutes; P=0.001). Additionally, the spillage rate of the LESS-I group was significantly greater than that of the LESS-E group (46.8% vs. 17.7%; P<0.001). The mean pain scores at 6 postoperative hours as measured by a visual analogue scale were significantly greater in the LESS-I group than the LESS-E group (3.85 vs. 3.37; P=0.016). CONCLUSIONS: LESS-E cystectomy is a safe and feasible approach with a shorter operation time and lower spillage rate than LESS-I cystectomy.

Molecular subtypes identified by gene expression profiling in early stage endometrioid endometrial adenocarcinoma.

BACKGROUND: Early stage (FIGO stage I-II) endometrioid endometrial adenocarcinoma (EEA) is very common in clinical practice. However, patients with the early stage EEA show various clinical behaviors due to biological heterogeneity. Hence, we aimed to discover distinct classes of tumors based on gene expression profiling, and analyze whether the molecular classification correlated with the histopathological stages or other clinical parameters. METHODS: Hierarchical clustering was performed for class discovery in 28 early stage EEA samples using a special cDNA microarray chip containing 492 genes designed for endometrial cancer. Correlations between clinicopathologic parameters and our classification were analyzed. And the significance analysis of microarrays (SAM) array was used to identify the signature genes according to the tumor grade and myometrial invasion. RESULTS: Three tumor subtypes (subtypes I, II and III) were identified by hierarchical clustering, each subtype had different clinicopathological factors, such as tumor grade, myometrial invasion status, and FIGO stage. Moreover, SAM analysis showed 34 up-regulated genes in high grade tumors, and 38 up-regulated genes and 1 down-regulated in deep myometrial invasive tumors. The overlap genes between these two high-risk factors were markedly up-regulated in subtype I, but down-regulated in subtype III. CONCLUSION: We have identified novel molecular subtypes in early stage EEA. Differential gene signatures characterize each tumor subtype, which could be used for recognizing the tumor risk and providing a basis for further treatment stratification.

Up-regulation of mitochondrial antioxidation signals in ovarian cancer cells with aggressive biologic behavior.

OBJECTIVE: Recently, a high frequency of mutations in mitochondrial DNA (mtDNA) has been detected in ovarian cancer. To explore the alterations of proteins in mitochondria in ovarian cancer, a pair of human ovarian carcinoma cell lines (SKOV3/SKOV3.ip1) with different metastatic potentials was examined. METHODS: Cancer cells SKOV3.ip1 were derived from the ascitic tumor cells of nude mice bearing a tumor of ovarian cancer cells SKOV3. SKOV3.ip1 exhibited a higher degree of migration potential than its paired cell line SKOV3. The proteins in the mitochondria of these two cells were isolated and separated by 2-D gel electrophoresis. The differently expressed proteins were extracted and identified using matrix assisted laser desorption ionisation/time-of-flight/time-of-flight (MALDI-TOF/TOF), and finally a selected protein candidate was further investigated by immunohistochemistry (IHC) method in nude mice bearing tumor tissues of these two cells. RESULTS: A total of 35 spots with different expressions were identified between the two cells using 2D-polyacrylamide gel electrophoresis (PAGE) approach. Among them, 17 spots were detected only in either SKOV3 or SKOV3.ip1 cells. Eighteen spots expressed different levels, with as much as a three-fold difference between the two cells. Twenty spots were analyzed using MALDI-TOF/TOF, and 11 of them were identified successfully; four were known to be located in mitochondria, including superoxide dismutase 2 (SOD2), fumarate hydratase (FH), mitochondrial ribosomal protein L38 (MRPL38), and mRNA turnover 4 homolog (MRTO4). An increased staining of SOD2 was observed in SKOV3.ip1 over that of SKOV3 in IHC analysis. CONCLUSIONS: Our results indicate that the enhanced antioxidation and metabolic potentials of ovarian cancer cells might contribute to their aggressive and metastatic behaviors. The underlying mechanism warrants further study.

Molecular classification of human endometrial cancer based on gene expression profiles from specialized microarrays.

OBJECTIVE: To investigate whether the molecular classification of endometrial cancer based on gene expression profiles can predict the biological behavior of the tumors and inform prognosis. METHODS: An array containing 492 genes was used to generate gene expression profiles from 35 tumor samples. A hierarchical cluster algorithm was used to compare gene expression patterns among the tumor samples. RESULTS: A cluster analysis revealed 3 distinct tumor clusters. A comparative analysis of tumor type, grade, FIGO stage, and depth of myometrial invasion revealed significant differences in grade and stage among the clusters, which appear to group tumors with specific clinical behaviors. Moreover, the cluster analysis initially revealed 2 clusters of differentially expressed genes. One contained 38 genes that were upregulated in most samples of the cluster representing the most advanced disease, and the other contained 27 genes that were upregulated in most samples of the cluster representing the least advanced disease. CONCLUSION: Molecular classification of endometrial cancer based on gene expression profiles obtained by designing specialized microarrays indicated a marked correspondence with the histologic features and clinical behavior of endometrial cancer tumors.