Association of Concomitant MRI-Determined Anterolateral Complex Injury With Quantitative Measurements of Altered Rotational Tibiofemoral Position on MRI in Patients With ACL Injury.

Background: Reduced graft failure rates have been reported after anterior cruciate ligament (ACL) reconstruction combined with anterolateral complex (ALC) augmentation. However, the preoperative diagnosis of concomitant ALC injury remains a clinical challenge. Purpose: To identify the altered rotational tibiofemoral position on magnetic resonance imaging (MRI) in ACL-injured patients with concomitant ALC injury. Study Design: Cross-sectional study; Level of evidence, 3. Methods: Based on the evaluation of ALC abnormalities on MRI scans by experienced surgeons, 123 patients with nonchronic (<3 months) ACL injury confirmed by arthroscopy were included. The patients were divided into 2 groups-an ALC-injured group (n = 57) and an ALC-intact group (n = 66). The altered rotational tibiofemoral position was evaluated and compared by quantitatively measuring internal rotational tibial subluxation (IRTS) and axial internal tibial rotation (ITRa) on MRI. Multivariate logistic regression and receiver operating characteristic (ROC) analyses were performed to identify the factors associated with concomitant MRI-determined ALC injury. Results: The ALC-injured group showed significantly increased IRTS (P < .001), ITRa (P < .001), lateral anterior tibial subluxation (ATS) (P < .001), and global ATS (GATS) (P = .002) compared with the ALC-intact group, while no significant difference in medial ATS (P = .810) was observed. A strong positive correlation was identified between IRTS and ITRa (rP = 0.809; P < .001). Multivariate analyses revealed that IRTS (P < .001) and GATS (P = .016) were associated factors for the presence of concomitant MRI-determined ALC injury. IRTS (area under the curve [AUC] = 0.734) was more strongly associated with the outcome than GATS (AUC = 0.658) in ROC analyses, suggesting a more significant internal rotational subluxation than anterior subluxation of the tibia. An IRTS threshold of 3.1 mm demonstrated a specificity of 84.2% for indicating the presence of concomitant MRI-determined ALC injury. Conclusion: The presence of concomitant MRI-determined ALC injury in ACL-injured patients was associated with a significant increase in IRTS and ITRa compared with those with intact ALC, indicating that these MRI measurements of the altered rotational tibiofemoral position could serve as potential quantifiable indicators for identifying concomitant ALC injury in clinical practice.

Acute abdomen caused by spontaneous rupture of degenerative hysteromyoma during pregnancy: A case report.

BACKGROUND: Hysteromyoma is not a rare tumor among pregnant women. During pregnancy, the symptoms caused by hysteromyoma can be improved through conservative treatment in most cases. However, in order to ensure the safety of mothers and children, surgeries are necessary in some special cases. CASE SUMMARY: We report a case of pregnancy complicated with hysteromyoma red degeneration. The patient had peritonitis after sudden abdominal pain during the 20th week of pregnancy. Laparoscopic exploration suggested rupture and bleeding of hysteromyoma, which were improved after drainage and an anti-inflammatory treatment. A cesarean section was performed after full term. This case shows the complications of rupture after red degeneration of hysteromyoma during pregnancy. CONCLUSION: We should be alert to rupture of hysteromyoma during pregnancy, and active laparoscopic exploration is essential to improve the prognosis of such patients.

Glucocorticoids promote lung metastasis of pancreatic cancer cells through enhancing cell adhesion, migration and invasion.

Glucocorticoids (GCs) are the important stress hormones and widely prescribed as drugs. Although stress has been suggested as a promoter of tumor progression, the direct influence of GCs on metastasis of tumor is not fully understood. Metastasis is a major cause of death in pancreatic cancer patients. In the present study, we investigated the effect of GCs on progression of pancreatic cancer and elucidated the underlying mechanism. It was found that GCs significantly promote cell adhesion, migration, and invasion of pancreatic cancer cells in vitro and their lung metastasis in vivo. Further mechanistic studies showed that GCs notably up-regulate the expression of a trans-membrane glycoprotein, mucin 1 (MUC1) and increase the activation of AKT. Inhibiting MUC1 expression not only attenuates the activation of AKT, but also significantly reduces the promoting effects of GCs on cell adhesion, migration, invasion, and lung metastasis of pancreatic cancer cells. Moreover, GCs not only significantly up-regulate expression of Rho-associated kinase 1/2 (ROCK1/2) and matrix metalloproteinase 3 and 7 (MMP3/7), but also activate ROCK2, which are also involved in the pro-migratory and pro-invasive effects of GCs in pancreatic cancer cells. Taken together, our findings reveal that GCs promote metastasis of pancreatic cancer cells through complex mechanism. MUC1-PI3K/AKT pathway, ROCK1/2 and MMP3/7 are involved in the promoting effect of GCs on cell migration, invasion and metastasis in pancreatic cancer cells. These results suggest the importance of reducing stress and GCs administration in patients with pancreatic cancer to avoid an increased risk of cancer metastasis.

Comparisons of diagnostic performance and the reliability in identifying ACL injury between two measuring protocols of anterior tibial subluxation on MR images.

OBJECTIVES: To evaluate the diagnostic performance in identifying an anterior cruciate ligament (ACL) injury and the reliability between two measuring protocols of anterior tibial subluxation (ATS). MATERIALS AND METHODS: A total of 165 patients with ACL injury and 157 ACL-intact patients were included in this study. Two different measuring protocols of ATS were performed on sagittal MR images, including the modified protocol using the longitudinal tibial axis (axis protocol) and the established protocol using a line perpendicular to the tibial plateau (plateau protocol). Receiver-operating characteristic (ROC) curves were calculated to evaluate the diagnostic performance in identifying an ACL injury, and areas under the curves (AUCs) were compared between the two protocols. Intra- and interobserver reliability tests were performed to evaluate the reliability of the measurements. RESULTS: Lateral ATS (P < 0.001) and medial ATS (P < 0.001) were increased in patients with ACL injury under both protocols. To identify an ACL injury, ATS measured under the axis protocol showed higher AUC values than the plateau protocol, including lateral ATS (AUC 0.828 vs. 0.688, P < 0.001), medial ATS (AUC 0.829 vs. 0.789, P = 0.013), and the combined indicator of lateral and medial ATS (AUC 0.885 vs. 0.810, P < 0.001). Reliability tests showed that both protocols were reliable. CONCLUSIONS: ATS measured under the modified protocol using the longitudinal tibial axis showed superior diagnostic performance in identifying an ACL injury compared to the established protocol, indicating that the modified protocol may better reflect the characteristics of an ACL-deficient knee.

Anterior tibial subluxation measured under a modified protocol is positively correlated with posterior tibial slope: a comparative study of MRI measurement methods.

PURPOSE: Anatomic factors, such as posterior tibial slope (PTS) and anterior tibial subluxation (ATS) obtained by quantitative measurement, have been proposed as predictors for clinical outcomes of anterior cruciate ligament (ACL) reconstruction. However, the correlation between PTS and ATS is controversial, and the method for quantitative ATS measurement remains unsettled. This study aimed to identify the correlation between PTS and ATS in patients with injured and intact ACLs and compare the two ATS measuring protocols. METHODS: This study included 128 ACL-injured and 176 ACL-intact patients with no concomitant ligament injuries. PTS and ATS were measured on sagittal MRI. ATS was measured using two measuring protocols, including the modified protocol using the longitudinal tibial axis (axis protocol) and the established protocol using a line perpendicular to the tibial plateau (plateau protocol). Correlation analyses between PTS and ATS and between PTS and the difference in the ATS value measured under the two protocols (ATSdiff) were performed. The difference between the two ATS measuring protocols was further analyzed by trigonometric analysis. Intra- and interobserver reliability tests were performed for the axis protocol. RESULTS: Under the axis protocol, ATS was positively correlated with PTS in both the ACL-injured and ACL-intact groups (p < 0.001). Under the plateau protocol, no correlation was observed in the ACL-injured group. In the ACL-intact group, no correlation was observed for lateral ATS, and a negative correlation was observed for medial ATS (p = 0.001). ATSdiff was positively correlated with PTS (p < 0.001), indicating that the two protocols varied greatly in those with a steep PTS. Trigonometric analysis showed that a steep PTS influenced the measurement of ATS under the plateau protocol but not the axis protocol. Intra- and interobserver reliability tests showed good-to-excellent strength of reliability for the ATS measurement under the axis protocol. CONCLUSION: ATS measured under the axis protocol was positively correlated with PTS, indicating that a steep PTS was associated with a worse anatomic tibiofemoral relationship. The axis protocol for ATS measurement is a promising method for clinical use since it is not influenced by PTS and reflects the global position of the tibia. LEVEL OF EVIDENCE: III.

Anterior and rotational tibial subluxation in the setting of anterior cruciate ligament injuries: An MRI analysis.

BACKGROUND: The relationship between preoperative tibiofemoral position and failure of anterior cruciate ligament (ACL) reconstruction has been widely discussed. Most established methods for measuring tibiofemoral position on magnetic resonance imaging (MRI) mainly focus on anterior tibial subluxation (ATS), while a quantitative measuring method for rotational tibial subluxation (RTS) is still undetermined. Moreover, there are still controversies about the related factors for ATS. The aim of this study was to quantitatively describe preoperative ATS and RTS in ACL-injured and ACL-intact knees and identify the related factors for ATS and RTS based on MRI images. METHODS: Demographic data and preoperative MRIs of 104 ACL-injured patients were retrospectively analyzed. ACL-intact knees were 1:1 matched as control group. ATS was measured using longitudinal tibial axis, and RTS was determined by the difference between lateral and medial ATS. Related factors for ATS and RTS were examined. RESULTS: Increased lateral ATS (P < 0.0001), medial ATS (P < 0.0001) and RTS (P = 0.0479) were observed in ACL-injured knees compared with the control group. Increased posterior tibial slope (PTS), Beighton Score ≥ 4, presence of meniscal injury and long injury-to-MRI time were identified as being correlated with the increase of ATS. Factors for the increase of RTS were increased lateral PTS, Beighton score ≥ 4, presence of lateral meniscal injury, and left side. CONCLUSIONS: In ACL-injured knees, tibia not only subluxated anteriorly in both lateral and medial compartments, but also rotated internally. During preoperative planning, attentions should be paid to the factors that are correlated with altered tibiofemoral position.

[Absence of BAX differentially affects astrocyte density in the mouse cortex and hippocampus].

Astrocytes are a heterogenous group of macroglia present in all regions of the brain and play critical roles in many aspects of brain development, function and disease. Previous studies suggest that the B-cell lymphoma-2 associated X protein (BAX)-dependent apoptosis plays essential roles in regulating neuronal number and achieving optimal excitation/inhibition ratio. The aim of the present paper was to study whether BAX regulates astrocyte distribution in a region-specific manner. Immunofluorescence staining of SOX9 was used to analyze and compare astrocyte density in primary somatosensory cortex, motor cortex, retrosplenial cortex and hippocampus in heterozygous and homozygous BAX knockout mice at age of six weeks when cortical development has finished and glia development has reached a relatively steady state. The results showed that astrocyte density varied significantly among different cortical subdivisions and between cortex and hippocampus. In contrast to the significant increase in GABAergic interneurons, the overall and region-specific astrocyte density remained unchanged in the cortex when BAX was absent. Interestingly, a significant reduction of astrocyte density was observed in the hippocampus of BAX knockout mice. These data suggest that BAX differentially regulates neurons and astrocytes in cortex as well as astrocytes in different brain regions during development. This study provided important information about the regional heterogeneity of astrocyte distribution and the potential contribution of BAX gene during development.

Influence of the universal two-child policy on obstetric issues.

OBJECTIVE: The universal two-child policy was implemented in January 2016 in China. The objective of this study was to compare the influence of change in fertility policy on obstetric issues. STUDY DESIGN: 2016 was taken as the cut-off point, and a retrospective study was conducted on data of patients who delivered in the China-Japan Friendship Hospital from January 2014 to December 2018. Maternal characteristics, mode of delivery, and pregnancy outcomes were studied in detail. RESULTS: Total 8931 babies were delivered from 2014 to 2018. There was a marked increase in the birth rate after the two-child policy. The percentage of elderly pregnant women and rate of cesarean sections increased significantly in 2017 and 2018. The primary cesarean section rates in 2017 and 2018 were significantly lower than those in 2014, 2015, and 2016. Increased incidence of placenta previa and postpartum hemorrhage were observed; however, no significant differences were seen in the rates of hypertensive disorders, gestational diabetes mellitus, and neonatal asphyxia within these five years. CONCLUSION: The implementation of two-child policy has changed the mode of obstetrics and has presented great challenges. With hierarchical management of high-risk patients and control of the cesarean section rate, we can ensure the safety of pregnant women and newborns.

Functional follow-up after endoscopic calcaneoplasty for Haglund's deformity using the biodex isokinetic muscle testing system: A case series.

Isokinetic muscle strength test implemented by the Biodex system is a method used for evaluating muscle function that has been applied clinically in the field of sports and rehabilitation medicine. However, information on its application on Haglund's deformity remain insufficient. Therefore, the present study examined the effectiveness of the muscle strength test using the Biodex system in evaluating the recovery of athletic capacity in patients with Haglund's deformity following endoscopic surgery. In total, 34 patients treated by the authors from June 2012 to November 2018 at Peking University Third Hospital (Beijing, China) were included. To compare muscle strength before surgery, then 3 and 6 months after surgery, using the uninjured side as the control, the Biodex system test was conducted in parallel to the collection of the American Orthopaedic Foot and Ankle Score values and visual analog scale scores. The Biodex system test results showed that Haglund's deformity mainly hinders plantar flexion strength. Patients recovered daily living capacity within 3 months and athletic capacity within 6 months following surgery, which matched the AOFAS values, VAS scores and the self-assessments of the patients. These findings suggest that the Biodex system can dynamically reflect the degree of postoperative recovery in Haglund's deformity.

MFGE8, ALB, APOB, APOE, SAA1, A2M, and C3 as Novel Biomarkers for Stress Cardiomyopathy.

BACKGROUND: Stress cardiomyopathy (SCM) is a transient reversible left ventricular dysfunction that more often occurs in women. Symptoms of SCM patients are similar to those of acute coronary syndrome (ACS), but little is known about biomarkers. The goals of this study were to identify the potentially crucial genes and pathways associated with SCM. METHODS: We analyzed microarray datasets GSE95368 derived from the Gene Expression Omnibus (GEO) database. Firstly, identify the differentially expressed genes (DEGs) between SCM patients in normal patients. Then, the DEGs were used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Finally, the protein-protein interaction (PPI) network was constructed and Cytoscape was used to find the key genes. RESULTS: In total, 25 DEGs were identified, including 10 upregulated genes and 15 downregulated genes. These DEGs were mainly enriched in ECM-receptor interaction, dilated cardiomyopathy (DCM), human papillomavirus infection, and focal adhesion, whereas in GO function classification, they were mainly enriched in the extracellular region, positive regulation of the multicellular organismal process, establishment of localization, and intracellular vesicle. CONCLUSION: Seven hub genes contained APOE, MFGE8, ALB, APOB, SAA1, A2M, and C3 identified as hub genes of SCM, which might be used as diagnostic biomarkers or molecular targets for the treatment of SCM.

The mechanisms and significance of up-regulation of RhoB expression by hypoxia and glucocorticoid in rat lung and A549 cells.

Small guanosine triphosphate (GTP)-binding protein RhoB is an important stress sensor and contributes to the regulation of cytoskeletal organization, cell proliferation and survival. However, whether RhoB is involved in the hypoxic response and action of glucocorticoid (GC) is largely unknown. In this study, we investigated the effects of hypoxia or/and GC on the expression and activition of RhoB in the lung of rats and human A549 lung carcinoma cells, and further studied its mechanism and significance. We found that hypoxia and dexamethasone (Dex), a synethic GC, not only significantly increased the expression and activation of RhoB independently but also coregulated the expresion of RhoB in vitro and in vivo. Up-regulation of RhoB by hypoxia was in part through stabilizing the RhoB mRNA and protein. Inhibiting hypoxia-activated hypoxia-inducible transcription factor-1α (HIF-1α), c-Jun N-terminal kinase (JNK) or extracellular signal-regulated kinase (ERK) with their specific inhibitors significantly decreased hypoxia-induced RhoB expression, indicating that HIF-1α, JNK and ERK are involved in the up-regulation of RhoB in hypoxia. Furthermore, we found that knockdown of RhoB expression by RhoB siRNA not only significantly reduced hypoxia-enhanced cell migration and cell survival in hypoxia but also increased the sensitivity of cell to paclitaxel (PTX), a chemotherapeutic agent, and reduced Dex-enhanced resistance to PTX-chemotherapy in A549 cells. Taken together, the novel data revealed that hypoxia and Dex increased the expression and activation of RhoB, which is important for hypoxic adaptation and hypoxia-accelerated progression of lung cancer cells. RhoB also enhanced the resistance of cell to PTX-chemotherapy and mediated the pro-survival effect of Dex.

A Marked Increase in Obstetric Hysterectomy for Placenta Accreta.

BACKGROUND: Obstetric hysterectomy (OH) as a lifesaving measure to manage uncontrolled uterine hemorrhage appears to be increasing recently. The objective of this study was to determine the etiology and changing trends of OH and to identify those at particular risk of OH to enhance the early involvement of multidisciplinary intensive care. METHODS: A retrospective study was carried out in patients who had OH in China-Japan Friendship Hospital from 2004 to 2014. Maternal characteristics, preoperative evaluation, operative reports, and prenatal outcomes were studied in detail. RESULTS: There were 19 cases of OH among a total of 18,838 deliveries. Comparing the study periods between 2004-2010 and 2011-2014, OH increased from 0.8/1000 (10/12,890) to 1.5/1000 (9/5948). Indications for OH have changed significantly during this study period with uterine atony decreasing from 50.0% (5/10) to 11.1% (1/9) (P < 0.05), and placenta accreta as the indication for OH has increased significantly from 20.0% (2/10) to 77.8% (7/9) (P < 0.05). Ultrasonography and magnetic resonance imaging (MRI) have been used to make an exact antepartum diagnosis of placenta accreta. A multidisciplinary management led to improved outcomes for patients with placenta accreta. CONCLUSION: As the multiple cesarean delivery rates have risen, there has been a dramatic increase in OH for placenta accreta. An advance antenatal diagnosis of ultrasonography, and MRI, and a multidisciplinary teamwork can maximize patients' safety and outcome.

The mechanism and significance of synergistic induction of the expression of plasminogen activator inhibitor-1 by glucocorticoid and transforming growth factor beta in human ovarian cancer cells.

Plasminogen activator inhibitor-1 (PAI-1) plays a key role in tissue remodeling and tumor development by suppression of plasminogen activator function. Glucocorticoids (GCs) and transforming growth factor beta (TGF-ß) signal pathways cross-talk to regulate gene expression, but the mechanism is poorly understood. Here we investigated the mechanism and significance of co-regulation of PAI-1 by TGF-ß and dexamethasone (DEX), a synthetic glucocorticoid in ovarian cancer cells. We found that TGF-ß and DEX showed rapidly synergistic induction of PAI-1 expression, which contributed to the early pro-adhesion effects. The synergistic induction effect was accomplished by several signal pathways, including GC receptor (GR) pathway and TGF-ß-activated p38MAPK, ERK and Smad3 pathways. TGF-ß-activated p38MAPK and ERK pathways cross-talked with GR pathway to augment the expression of PAI-1 through enhancing DEX-induced GR phosphorylation at Ser211 in ovarian cancer cells. These findings reveal possible novel mechanisms by which TGF-ß pathways cooperatively cross-talk with GR pathway to regulate gene expression.

Haplotype-based approach for noninvasive prenatal tests of Duchenne muscular dystrophy using cell-free fetal DNA in maternal plasma.

PURPOSE: This study demonstrates noninvasive prenatal testing (NIPT) for Duchenne muscular dystrophy (DMD) using a newly developed haplotype-based approach. METHODS: Eight families at risk for DMD were recruited for this study. Parental haplotypes were constructed using target-region sequencing data from the parents and the probands. Fetal haplotypes were constructed using a hidden Markov model through maternal plasma DNA sequencing. The presence of haplotypes linked to the maternal mutant alleles in males indicated affected fetuses. This method was further validated by comparing the inferred single-nucleotide polymorphism (SNP) genotypes to the direct sequencing results of fetal genomic DNA. Prenatal diagnosis was confirmed with amniocentesis, and those results were interpreted in a blinded fashion. RESULTS: The results showed an average accuracy of 99.98% for the total inferred maternal SNPs. With a mean depth of 30× achieved in the 10-Mb target region of each sample, the noninvasive results were consistent with those of the invasive procedure. CONCLUSION: This is the first report of NIPT for DMD and the first application of a haplotype-based approach in NIPT for X-linked diseases. With further improvements in accuracy, this haplotype-based strategy could be feasible for NIPT for DMD and even other X-linked single-gene disorders.

Differential patterns of insulin secretion and sensitivity in patients with type 2 diabetes mellitus and nonalcoholic fatty liver disease versus patients with type 2 diabetes mellitus alone.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) often coexist and have adverse outcomes. The aim of our study was to elucidate metabolic abnormalities in patients with DM-NAFLD versus those with T2DM alone. METHODS: Patients were divided into two groups: 26 T2DM patients with NAFLD and 26 gender-, age-, and body mass index-matched patients with T2DM alone. Patients took a 75-g oral glucose tolerance test (OGTT), which measured serum insulin and C-peptide (C-p) levels at baseline (0 min), 30 min, 60 min, and 120 min after glucose challenge. RESULTS: Patients with DM-NAFLD or T2DM alone had similar blood glucose levels. ß-cell hypersecretion was more obvious in patients with DM-NAFLD. In addition, fasting, early-phase, and late-phase C-peptide levels were significantly increased in patients with DM-NAFLD (ΔC-p 0-30 min, P < 0.05; Area Under the Curve (AUC) C-p/PG 30-120 min ratio, P < 0.01; and AUC C-p 30-120 min, P < 0.01). Hepatic and extrahepatic insulin resistance during the OGTT did not differ significantly between groups. Hepatic insulin sensitivity independently contributed to the early phase (0-30 min) of the OGTT in patients with T2DM and NAFLD, whereas a significant deficit in late insulin secretion independently contributed to the 30-120 min glucose status in patients with T2DM only. CONCLUSIONS: In patients with similar levels of insulin resistance and hyperglycemia, DM-NAFLD was associated with higher serum insulin levels than T2DM alone. Hyperinsulinemia is caused mainly by ß-cell hypersecretion. The present study demonstrates pathophysiological differences in mechanisms of insulin resistance in patients with DM-NAFLD versus T2DM alone.

Overexpression of claudin-4 may be involved in endometrial tumorigenesis.

To clarify the role of claudin-4 in endometrial tumorigenesis and to explore whether claudin-4 could be a potentially useful agent in the treatment of endometrial carcinoma, the expression of claudin-4 in endometrial carcinoma was investigated. The relationship between therapy with anti-neoplastic agents and the expression of claudin-4 was also analyzed using an endometrial carcinoma xenograft model. The expression of claudin-4 in endometrial endometrioid adenocarcinoma (EEC) and normal human endometrial tissue was determined using immunohistochemistry and real-time PCR. Ninety female BALB/c nu/nu mice were transplanted with Ishikawa endometrial cancer cells. The mice were divided into three groups with different intraperitoneal treatments: cisplatin, paclitaxel or saline solution. After the observation period tumors were extracted and stained with monoclonal antibody against claudin-4. The mRNA expression of claudin-4 was also detected using real-time PCR. Expression of claudin-4 was significantly increased at both protein and mRNA levels in the EEC group compared with the group of normal cyclic endometrium. In the study of Ishikawa xenografts, no significant changes in tumor volume and claudin-4 expression were shown in the paclitaxel group compared with the control group. A significant reduction of tumor growth and a significant decrease in claudin-4 expression were observed in the cisplatin group. These results demonstrate that claudin-4 is strongly expressed in EEC. Claudin-4 is a useful biomarker in the treatment of patients with endometrial carcinoma.

[Study of claudin-4 in the diagnosis and treatment of endometrial carcinoma].

OBJECTIVE: To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma. METHODS: The expression of claudin-4 in 62 endometrioid endometrial carcinoma (EEC), 30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and real-time PCR. Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells, which were divided into three groups with different intraperitoneal treatments with cisplatin, paclitaxel and saline solution. After the observation period, the tumors were extracted and stained with monoclonal antibody against claudin-4. The messenger RNA expression of claudin-4 was also detected using real-time PCR. RESULTS: Among the EEC samples, 34% (21/62) showed medium staining for claudin-4 and 66% (41/62) showed intense staining. In atypical hyperplasia group, 27% (8/30) showed weak staining, 53% (16/30) showed medium staining and 20% (6/30) showed intense staining for claudin-4. Of the normal endometrial tissue, 47% (28/60) showed weak staining and 53% (32/60) showed no staining for claudin-4. According to real-time PCR, the relative quantity of claudin-4 was 170 ± 12 in EEC group, 89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium. Compared with those in atypical hyperplasia group and normal endometrium group, the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P < 0.05). In the study of Ishikawa xenografts, no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group. Nevertheless, a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group. After cisplatin treatment, the tumor volume was significantly decreased [(0.51 ± 0.21) versus (0.73 ± 0.12) cm(3)], and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27). CONCLUSION: These results demonstrate that claudin-4 is strongly expressed in EEC, which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

[Regulation of claudin-4 gene expression in endometrial adenocarcinoma Ishikawa cell line by progesterone].

OBJECTIVE: To explore the regulation of claudin-4 expression in endometrial adenocarcinoma cell lines by progesterone. METHODS: Ishikawa cells were treated with various concentrations of megestrol acetate (MA: 2, 5, 10, 15, 20 mg/L). After cultured for 24, 48 and 72 hours, cells growth were measured by methyl thiazolyl tetrazolium (MTT). The group of Ishikawa cells incubated with MA at the 50% inhibitory concentration (IC(50)) was selected for cell apoptosis assay by using transmission electron microscopy and flow cytometry method. Real-time PCR and western blot were used for detecting the mRNA and protein expression levels of claudin-4. The localization of claudin-4 was examined by immunofluorescent staining. RESULTS: The inhibitory effects of megestrol acetate on the growth of Ishikawa cells were dose-dependent and time-dependent. IC(50) of MA on Ishikawa cells was 15 mg/L after incubated for 72 hours. After MA treatment, Ishikawa cells showed shrinkage, nuclear chromatin condensation, fractures of nuclear membrane and endoplasmic reticulum expansion, even round apoptotic bodies were found. The apoptosis rate of cells before MA treatment was (0.076 ± 0.024)%, and the rate was (3.934 ± 0.816)% by MA treated for 72 hours, in which there were signicant difference (P < 0.05). The relative quantification of claudin-4 mRNA and protein of the cells before MA treatment were 0.64 ± 0.20 and 0.94 ± 0.18, while they were 0.47 ± 0.15 and 0.62 ± 0.15 after MA treated. The expression of claudin-4 was significantly decreased after MA treatment (P < 0.05). The localization of claudin-4 transferred from cytomembrane to cytoplasm and nucleus after MA treatment. CONCLUSIONS: MA could inhibite the growth of Ishikawa cells, in which the mechanism may be decrease the expression of claudin-4 and the apoptosis of cells. The distribution change of claudin-4 may be related to the anti-cancer effect of progesterone.

Feasibility of laparoscopic extraperitoneal pelvic lymphadenectomy in gynecologic malignancies.

OBJECTIVE: The aim of this study is to evaluate the feasibility of laparoscopic extraperitoneal pelvic lymphadenectomy (LEPL) in gynecologic malignancies. METHODS: Twenty-nine women with cervical, ovarian or endometrial cancer underwent laparoscopic extraperitoneal pelvic lymphadenectomy between July 2008 and December 2010. The operating time, nodal yield, blood loss and complications were recorded. RESULTS: The number of patients with cervical, ovarian and endometrial carcinoma was 14, 3 and 12, respectively. The median age of patients was 48.9±12.6 years. The median body mass index was 25.6±4.8. Conversion to the transperitoneal laparoscopic approach was necessary in 6 patients for peritoneal tears causing CO(2) gas leakage. Among the remaining 23 patients, the median operating time for laparoscopic extraperitoneal pelvic lymphadenectomy was 69 min (range 50-126 min), and the median estimated blood loss was 20 ml (range 5-105 ml). The median total number of resected nodes was 26 (range 14-42), and complications related to the procedure were rare. CONCLUSIONS: Laparoscopic extraperitoneal pelvic lymphadenectomy is a feasible and safe procedure. It can be used in gynecologic malignancies.

Altered expression of claudin-3 and claudin-4 in ectopic endometrium of women with endometriosis.

OBJECTIVE: To investigate the expression of claudin-3 and claudin-4 in the eutopic and ectopic endometrium of women with endometriosis and to evaluate the role of claudin-3 and claudin-4 in the pathogenesis of endometriosis. DESIGN: Cross-sectional measurement of gene expression levels of claudin-3 and claudin-4 on endometriotic tissue. SETTING: Academic. PATIENT(S): Thirty-five patients with endometriosis and 35 healthy women who were free of endometriosis were recruited for the study. INTERVENTION(S): Expression of claudin-3 and claudin-4 were investigated with immunohistochemical analysis, Western blot, and real-time polymerase chain reaction. Morphologic change of tight junction was also observed in different kinds of endometria. MAIN OUTCOME MEASURE(S): The expression levels of claudin-3 and claudin-4 in epithelial cells from 35 ectopic endometrial tissues, 27 eutopic endometrial tissues from women with endometriosis, and 35 normal endometrial tissues from women without endometriosis. RESULT(S): Expression of claudin-3 and claudin-4 was significantly lower in the ectopic endometriotic tissue than in the eutopic endometrium from women with endometriosis and normal controls at both the messenger RNA and protein levels. No significant difference was found between eutopic endometrium from women with endometriosis and normal endometrium from women without endometriosis. CONCLUSION(S): Down-regulated expression of claudin-3 and claudin-4 in ectopic endometrium suggests that claudin-3 and claudin-4 might play a pathogenic role in the formation of endometriosis.