Gene Set-Based Functionome Analysis of Pathogenesis in Epithelial Ovarian Serous Carcinoma and the Molecular Features in Different FIGO Stages.

Serous carcinoma (SC) is the most common subtype of epithelial ovarian carcinoma and is divided into four stages by the Federation of Gynecologists and Obstetrics (FIGO) staging system. Currently, the molecular functions and biological processes of SC at different FIGO stages have not been quantified. Here, we conducted a whole-genome integrative analysis to investigate the functions of SC at different stages. The function, as defined by the GO term or canonical pathway gene set, was quantified by measuring the changes in the gene expressional order between cancerous and normal control states. The quantified function, i.e., the gene set regularity (GSR) index, was utilized to investigate the pathogenesis and functional regulation of SC at different FIGO stages. We showed that the informativeness of the GSR indices was sufficient for accurate pattern recognition and classification for machine learning. The function regularity presented by the GSR indices showed stepwise deterioration during SC progression from FIGO stage I to stage IV. The pathogenesis of SC was centered on cell cycle deregulation and accompanied with multiple functional aberrations as well as their interactions.

Gene Set-Based Integrative Analysis Revealing Two Distinct Functional Regulation Patterns in Four Common Subtypes of Epithelial Ovarian Cancer.

Clear cell (CCC), endometrioid (EC), mucinous (MC) and high-grade serous carcinoma (SC) are the four most common subtypes of epithelial ovarian carcinoma (EOC). The widely accepted dualistic model of ovarian carcinogenesis divided EOCs into type I and II categories based on the molecular features. However, this hypothesis has not been experimentally demonstrated. We carried out a gene set-based analysis by integrating the microarray gene expression profiles downloaded from the publicly available databases. These quantified biological functions of EOCs were defined by 1454 Gene Ontology (GO) term and 674 Reactome pathway gene sets. The pathogenesis of the four EOC subtypes was investigated by hierarchical clustering and exploratory factor analysis. The patterns of functional regulation among the four subtypes containing 1316 cases could be accurately classified by machine learning. The results revealed that the ERBB and PI3K-related pathways played important roles in the carcinogenesis of CCC, EC and MC; while deregulation of cell cycle was more predominant in SC. The study revealed that two different functional regulation patterns exist among the four EOC subtypes, which were compatible with the type I and II classifications proposed by the dualistic model of ovarian carcinogenesis.

[Bio-mechanical evaluation of extreme lateral transforaminal lumbar interbody fusion].

OBJECTIVE: To evaluate the feasibility and post-operation stability of extreme lateral transforaminal lumbar interbody fusion (E-TLIF) and other traditional surgical approach via bio-mechanical test. METHODS: There were 24 normal lumbar spine segment of swine were divided into the following four groups: control group, standard group (internal fixed with pedicle screws only), transforaminal lumbar interbody fusion (TLIF) group and E-TLIF group. The specimen in anteflect, hypsokinesis, lateral flexion and rotate movements were tested respectively with bio-mechanical devices to study on the load-straining changes and biomechanics index. RESULTS: After TLIF or E-TLIF, specimen turned out more steady than normal control group (t = 4.17 - 4.53, P < 0.01). Compared with TLIF group [linear displacement (3.98 ± 0.22) mm, angular displacement 3.03° ± 0.18°], specimen after E-TLIF [linear displacement (3.40 ± 0.09) mm, angular displacement 2.57° ± 0.12°] were more stable in biomechanics index on lateral flection movement (t = 2.61, P < 0.05), but no difference on axial or rotational movements (P > 0.05). CONCLUSION: E-TLIF is a safe and more efficient operation approach.

Characterization of waste cell biomass derived glutamate decarboxylase for in vitro γ-aminobutyric acid production and value-addition.

Waste biomass of Lactobacillus brevis obtained from in vivo γ-aminobutyric acid (GABA) production was used for value-addition. This study aims to extract glutamate decarboxylase (GAD) and characterize it for in vitro GABA production. Extracted GAD showed an excellent activity for in vitro GABA production. 52 W ultrasonic output was best in crude GAD extraction which was purified by Q HP anion-exchange column followed by Superdex-200 colloid separation column. The molecular weight of the purified GAD was determined to be ~53 kDa, and the Km value for L-glutamic acid was calculated ~7.65 mM. Pyridoxal 5'-phosphate (PLP) acted as the best cofactor for GAD. Optimum temperature and PLP dosing were deferring for crude and purified enzyme forms which respectively exhibited at 45°C, 55°C, 200 µmol and 20 µmol whereas optimum pH was the same at 4.5. GAD finds applications in food industries hence its detailed characterization would be promising for commercial exploitations.

Small supernumerary marker chromosome originating from chromosome 10 associated with an apparently normal phenotype.

Small supernumerary marker chromosomes (sSMC) originating from chromosome 10 are rare. Only seven cases have been documented, and among those three cases were diagnosed prenatally. We reported on another prenatal diagnosis of a de novo mosaic sSMC in an apparently normal female fetus whose mother had conceived with assisted reproductive technology (ART) procedures. G-banding analysis of amniotic cells was performed. Spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) studies with chromosome 10-specific alphoid satellite DNA probe were used to identify the chromosome 10 origin of the sSMC. Further FISH study with telomeric sequence probes showed that the sSMC lacked a hybridization signal, suggesting that the marker could be a ring chromosome. FISH studies using BAC clone probes specific for the regions within 10p11.2, 10q11.1, and 10q11.2 showed that the short arm breakpoint was located between 29.8 and 30.7 Mb from the 10p telomere, and that the long arm breakpoint was located less than 43.6 Mb from the 10p telomere. The karyotype of the fetus was 47,XX,+mar. ish der(10)(SKY+ CEP 10+, CTD-2130I7+, RP11-89J23-)/46,XX. Oligonucleotide microarray-based copy number variations (CNV) analysis was also performed and showed a 6.7 Mb duplication from 10p11.2 to 10q11.2 (36.2-42.9 Mb) with Affymetrix SNP-array 6.0 genotype: arr cgh. 10p11.2q11.2(CN_519687 --> CN_541524) X 3. At the 1-year follow-up, the baby did not have any findings of the trisomy 10p syndrome. This observation provided further credence to the concept that additional chromosome material of proximal 10p11.2 may not contribute to the trisomy 10p syndrome phenotype.

Low plasma levels of decoy receptor 3 (DcR3) in the third trimester of pregnancy with preeclampsia.

OBJECTIVE: The pathophysiology of preeclampsia, a major threat during pregnancy characterized by excessive inflammatory status, remains unclear. Decoy receptor 3 (DcR3), a soluble member of the tumor necrosis factor receptor (TNFR) superfamily, is capable of inducing anti-apoptosis via binding with TL1A and anti-inflammation by driving Th2 immune reactions. DcR3 may, therefore, play a role in immune modulation during pregnancy. The purpose of this study is to explore the role of DcR3 in normal and preeclamptic pregnancies. MATERIALS AND METHODS: Plasma samples from 104 normal pregnant women (26, 42, and 36 in the first, second, and third trimester, respectively) and 10 patients with preeclampsia in the third trimester were collected. Plasma DcR3 levels were determined by using commercial ELISA kits. ANOVA and linear regression analysis were performed to analyze the relationship between gestational age and DcR3 levels. After adjusting for gestational days, the levels of plasma DcR3 in preeclamptic and non-preeclamptic women in the third trimester were compared. RESULTS: The plasma levels of DcR3 gradually decreased as the gestational days increased during pregnancy (p < 0.05). In the third trimester, pregnant women with preeclampsia had significantly lower plasma DcR3 levels compared to non-preeclamptic women (p < 0.05). CONCLUSIONS: We found that plasma DcR3 levels gradually decreased as gestation progressed. The levels of plasma DcR3 in preeclamptic women were significantly lower than those of normal pregnant women, suggesting that a potential involvement of DcR3 in normal pregnancy and decreased levels of DcR3 may be related to preeclampsia.

Hyperbaric oxygen therapy for cesarean section wound in diabetes mellitus gravida.

We report the use of hyperbaric oxygen (HBO) therapy to treat the complication of necrotizing fasciitis following Cesarean section in a postpartum gravida with diabetes mellitus. Our patient was a 25-year-old, gravida 1, para 1, woman with a history of type 1 diabetes mellitus since the age of 18. The patient experienced preterm labor at 31+1 gestational weeks and was treated with magnesium sulfate for tocolytic therapy. The patient then went into labor at 39+6 gestational weeks. She received Cesarean section due to prolonged labor associated with non-reassuring fetal status of both smooth baseline and fetal tachycardia. An ultrasound scan of the lower abdomen on the 4th postoperative day revealed fluid collection measuring 4 mm over the rectus fascia and edematous change of the surrounding soft tissues under the Cesarean section incision site. The patient eventually received HBO for a total of 7 days. Following HBO, the condition of the surgical wound improved dramatically. The results of this case showed that HBO has the potential to be a cost-effective way to enhance the healing of necrotizing fasciitis in diabetes mellitus gravida.

Lamivudine therapy in the treatment of chronic hepatitis B with acute exacerbation during pregnancy.

We report a case of chronic hepatitis B carrier gravida who had acute exacerbation during pregnancy. She had been taking lamivudine 100 mg/qd for 17 months when hepatitis B virus (HBV) DNA in the YMDD region of the polymerase gene (YMDD motif) mutant was noted. After discontinuing lamivudine, she became pregnant. HBeAg became positive again and liver enzymes were elevated during the first trimester of pregnancy. She received the hepatoprotective agent silymarin 150 mg bid at 13+2 gestational weeks. Serum aspartate aminotransferase (AST) dropped to 757 U/L at 15+0 gestational weeks, but serum alanine aminotransferase (ALT) flared up to 2,230 U/L and AST to 2,250 U/L at 17+1 gestational weeks. Serum HBV-DNA test revealed serum HBV-DNA concentration of 7.31 x 10(8) copies/mL. Lamivudine 100 mg/qd and silymarin 150 mg/bid were initiated at 17+1 gestational weeks. Liver function showed gradual decline to ALT 341 U/L and AST 91 U/L at 21+0 gestational weeks, while HBeAg(+) converted to (-) and anti-HBe(-) converted to (+). Further treatment with lamivudine 100 mg/qd continued for 3 months. Serum HBV-DNA concentrations decreased to 3.19 x 10(2) copies/mL at 36+6 gestational weeks. Spontaneous delivery of a male baby weighing 3314 g occurred at 38+3 gestational weeks. The neonatal physical check-up revealed no congenital anomalies, and fetal growth was within normal reference ranges, suggesting that lamivudine may be safely used in the treatment of chronic hepatitis B with acute exacerbation during the second trimester of pregnancy.

Prenatal diagnosis of pulmonary sequestration by ultrasound and magnetic resonance imaging.

A 36-year-old multigravida, G2P1, underwent routine ultrasound scan at 22+1 weeks of gestation, which revealed a single normally growing fetus with left intrathoracic mass and left displacement of the cardiac apex. The left intrathoracic wedge-shaped hyperechogenic mass, measuring 32 x 25 mm in size, was situated at the lower portion of the left lung. A combination of color and power Doppler ultrasound allowed visualization of a vessel arising from the descending aorta, which supplied the mass. The diagnosis of extralobar pulmonary sequestration was made. Magnetic resonance imaging (MRI) was also performed and revealed a well-defined mass with homogeneous high-signal intensity when compared with normal lung tissue in the left upper lung field, which was compatible with pulmonary sequestration. The pulmonary mass was followed up by color and power Doppler every 2 weeks. The peak velocity of 11.85 cm/sec and the diameter of the feeding artery of 1.19 mm gradually decreased and disappeared 8 weeks later. The intrathoracic mass disappeared 10 weeks later at 32+1 gestational weeks. Repeat MRI also revealed spontaneous regression of the mass in favor of resorption of sequestration. The fetus was delivered at 38+1 gestational weeks. A male newborn weighing 2,520 g was spontaneously delivered with an Apgar score of 8 at 1 minute and 9 at 5 minutes. In our patient, it is suggested that progressive decreases in the peak velocity of the feeding vessel heralded the spontaneous regression of pulmonary sequestration not associated with hydrops/hydrothorax.

Prenatal diagnosis of dextrotransposition of the great arteries.

Dextrotransposition of the great arteries (DTGA) is a common cardiac cause of cyanosis in newborn infants that can cause acidosis and death within a short period of time unless there is a large atrial-level shunt or a patent ductus arteriosus. Here, we report a case of prenatal diagnosis of DTGA at 24+1 gestational weeks. In a tilted 4-chamber view, the pulmonary trunk branched to the left and the right pulmonary, with its root connected to the left ventricle outflow tract. In the short-axis view, the pulmonary trunk was shown to be parallel with the ascending aortic root. Cesarean section was performed due to the nonreassuring fetal status at 38+5 gestational weeks. The male neonate appeared to have mild cyanotic symptoms and weighed 3,108 g. Apgar scores were 8 and 9 at 1 and 5 minutes, respectively. Neonatal echocardiography was performed immediately after birth and the findings confirmed DTGA associated with atrial septal defect secundum. Postnatally, angiography confirmed the echocardiographic diagnosis of DTGA with a large atrial septal defect secundum and a large patent ductus arteriosus. Jatene arterial switch operation and atrial septal defect closure with Gore-Tex patch were performed. The neonate withstood the operation well and was discharged 27 days after birth weighing 2,950 g and in a stable condition. Prenatal diagnosis of DTGA can greatly aid to prepare the patient's family and the surgeon and significantly improve the outcome of complex heart disease in the neonatal period.

Magnetic resonance imaging as a diagnostic tool in pregnancy with appendiceal abscess.

Abdominal pain presenting itself during pregnancy may be multifactorial, requiring immediate attention and care. In cases of intractable pain without obstetrical condition, surgical abdominal exploration is widely advised. However, we present a case where a 30-year-old, gravida 1, para 0, female complained of persistent right abdominal pain during her 25th week of pregnancy. Ultrasound revealed a right upper quadrant cystic mass and magnetic resonance imaging was arranged with compatible findings. Final impression of appendiceal abscess was determined. Broad-spectrum antibiotics were administered and the patient was discharged in stable condition after 10 days of conservative treatment. She delivered a healthy baby boy at her 40th week of gestation uneventfully.

Fertility preservation with treatment of immature teratoma of the ovary.

Fertility preservation for a patient with advanced immature teratoma of the ovary is reported. The patient, a 29-year-old woman, delivered a healthy baby after having had ovarian immature teratoma, grade 3, uncertain stage, at 13 years of age. She was initially treated with unilateral salpingo-oophorectomy and a contralateral wedge resection for tumor invasion, followed by a 6-course cisplatin+vinblastine+bleomycin regimen, a second operation, and an additional 6-course etoposide and cisplatin regimen with complete remission. The patient delivered a healthy baby 16 years after the initial treatment. Based on this successful case, intensive fertility-preserving surgery followed by chemotherapy, even in advanced-stage immature teratomas of the ovary, may be effective in preserving the reproductive function of women with malignant immature teratomas of the ovary.

Dynamic alterations of the levels of tumor necrosis factor-α, interleukin-6, and interleukin-1ß in rat primary motor cortex during transhemispheric functional reorganization after contralateral seventh cervical spinal nerve root transfer following brachial plexus avulsion injuries.

The transfer of a contralateral healthy seventh cervical spinal nerve root (cC7) to the recipient nerve in the injured side is considered a reliable and effective procedure for restoration of the physiological functions of an injured hand after brachial plexus root avulsion injury (BPAI). Growing evidence shows that the transhemispheric cortical reorganization is induced after cC7 nerve transfer surgery. However, little is known about the underlying molecular mechanism. Proinflammatory cytokines reportedly play an important role in the neural plasticity. We hypothesize that proinflammatory cytokines are involved in the transhemispheric functional reorganization after cC7 transfer. In the present study, we investigated the level of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in the rat primary motor cortex after cC7 transfer following BPAI by enzyme-linked immunosorbent assay. The results showed that, in the sham group, no statistical significance was observed between the level of TNF-α, IL-6, and IL-1ß at each time point after the operation compared with that at day 0, respectively. However, in the unrepaired and repaired groups, the level of TNF-α, IL-6, and IL-1ß changed dynamically. The study is the first to provide evidence for the involvement of proinflammatory cytokines in transhemispheric functional reorganization after cC7 transfer following BPAI, which are useful for understanding the underlying mechanism.

Sphingosine-1-phosphate improves endothelialization with reduction of thrombosis in recellularized human umbilical vein graft by inhibiting syndecan-1 shedding in vitro.

Sphingosine-1-phosphate (S1P) has been known to promote endothelial cell (EC) proliferation and protect Syndecan-1 (SDC1) from shedding, thereby maintaining this antithrombotic signal. In the present study, we investigated the effect of S1P in the construction of a functional tissue-engineered blood vessel by using human endothelial cells and decellularized human umbilical vein (DHUV) scaffolds. Both human umbilical vein endothelial cells (HUVEC) and human cord blood derived endothelial progenitor cells (EPC) were seeded onto the scaffold with or without the S1P treatment. The efficacy of re-cellularization was determined by using the fluorescent marker CellTracker CMFDA and anti-CD31 immunostaining. The antithrombotic effect of S1P was examined by the anti-aggregation tests measuring platelet adherence and clotting time. Finally, we altered the expression of SDC1, a major glycocalyx protein on the endothelial cell surface, using MMP-7 digestion to explore its role using platelet adhesion tests in vitro. The result showed that S1P enhanced the attachment of HUVEC and EPC. Based on the anti-aggregation tests, S1P-treated HUVEC recellularized vessels when grafted showed reduced thrombus formation compared to controls. Our results also identified reduced SDC1 shedding from HUVEC responsible for inhibition of platelet adherence. However, no significant antithrombogenic effect of S1P was observed on EPC. In conclusion, S1P is an effective agent capable of decreasing thrombotic risk in engineered blood vessel grafts. STATEMENT OF SIGNIFICANCE: Sphingosine-1phosphate (S1P) is a low molecular-weight phospholipid mediator that regulates diverse biological activities of endothelial cell, including survival, proliferation, cell barrier integrity, and also influences the development of the vascular system. Based on these characters, we the first time to use it as an additive during the process of a small caliber blood vessel construction by decellularized human umbilical vein and endothelial cell/endothelial progenitor. We further explored the function and mechanism of S1P in promoting revascularization and protection against thrombosis in this tissue engineered vascular grafts. The results showed that S1P could not only accelerate the generation but also reduce thrombus formation of small caliber blood vessel.

Onset of systemic lupus erythematosus during pregnancy.

When systemic lupus erythematosus (SLE) is first suspected during pregnancy, though rare, the diagnostic criteria are not different from those for nonpregnant women. The pregnancy outcome is good if treatment with adequate immunosuppressive agents starts as soon as the diagnosis is made. There are 4 cases in this report who had SLE onset during pregnancy. Although 2 of them suffered from preeclampsia, all 4 pregnancies resulted in favorable outcomes after the lupus was controlled by medical treatment.

[Scanning aneugen and clastogen by micronuclei analysis using flow cytometry].

OBJECTIVE: To explore a flow cytometry (FCM)-based method for discriminating aneugen- or clastogen-induced micronuclei. METHODS: Cells were stained with anti-CD71-FITC and PI, and the PI fluorescent signal intensity of micronucleated reticulocyte (MN-RET) in the peripheral blood of NIH mouse treated with COL or CP was detected by flow cytometry. RESULTS: The ratio of the median of the intensity of MN-RET fluorescent signals to that of nucleated cell was low in the cyclophosphamide treated mouse, while the median was high in the colchicine treated mouse. CONCLUSION: The flow cytometry-based micronucleus assay can be used to discriminate primarily smaller MN induced by the clastogen exposure from the larger MN induced by an aneugen.

Gene set-based integrative analysis of ovarian clear cell carcinoma.

OBJECTIVE: The pathogenesis of ovarian clear cell carcinoma is still poorly understood; therefore, we conducted a gene set-based analysis by integrating datasets downloaded from publicly available microarray gene expression databases to investigate the pathogenesis of clear cell carcinoma, which was based on the regularity of functions defined by gene ontology or canonical pathway databases. MATERIALS AND METHODS: The gene expression profiles of 80 clear cell carcinomas and 136 normal ovarian controls were downloaded from the National Center for Biotechnology Information Gene Expression Omnibus database. The gene expression profiles were converted to the gene set regularity (GSR) indexes computed using the modified differential rank conservation, an algorithm measuring the degree of gene expression ranking change in a gene set. Then the differences of GSR indexes between clear cell carcinomas and normal ovarian controls were analyzed. RESULTS: Machine learning can accurately recognize and classify the patterns of functional regularities containing the GSR indexes between the clear cell carcinomas and normal controls with an accuracy of 99.3%. The significant aberrations included oxidoreductase activity, binding, transport, channel activity, cell adhesion, immune response, chromosome assembly, and the deregulated signaling molecules, such as guanyl nucleotide exchange factors, phosphoinositide 3-kinase-activating kinase, receptor tyrosine kinase B, and protein tyrosine kinase. CONCLUSION: Our pioneering works using the functionome, which was converted from microarray gene expression profiles for integrative analysis, showed a clear distinction of functional changes between the clear cell carcinomas and normal ovarian controls. This approach might provide a comprehensive view of the deregulated functions of clear cell carcinomas for further investigation.

Quantitative analysis of normal fetal medulla oblongata volume and flow by three-dimensional power Doppler ultrasound.

OBJECTIVE: Assessment of the fetal medulla oblongata volume (MOV) and blood flow might be important in the evaluation of fetal brain growth. We used three-dimensional power Doppler ultrasound (3DPDUS) to assess the fetal MOV and blood flow index in normal gestation. The relationships between these parameters were further analyzed. METHODS: We assessed the total volume and blood flow index of the fetal MO in normal pregnancies using a 3DPDUS (Voluson 730 Expert). The true sagittal plane over the fetal occipital area was measured by a 3D transabdominal probe to scan the fetal MO under the power Doppler mode. Then, we quantitatively assessed the total volume of the fetal MOV, mean gray area (MG), vascularization index (VI), and flow index (FI). RESULTS: A total of 106 fetuses, ranging from 19 weeks to 39 weeks of gestation, were involved in our study. The volume of the fetal MO was highly positively correlated with gestational age [correlation coefficient (r) = 0.686, p < 0.0001]. The MG was negatively correlated with gestational age [r = -0.544, p < 0.0001). VI and FI showed no significant correlation with gestational age (p = 0.123 and p = 0.219, respectively). CONCLUSION: 3DPDUS can be used to assess the fetal MOV and blood flow development quantitatively. Our study indicated that fetal MOV and blood flow correlated significantly with the advancement of gestational age. This information may serve as reference data for further studies of the fetal brain and blood flow under abnormal conditions.

Vaginal birth after cesarean section: 10 years of experience in a tertiary medical center in Taiwan.

OBJECTIVE: Because of the increased risk of uterine rupture and other morbidities, instances of trial of labor after cesarean (TOLAC) have decreased in number each year. Nevertheless, under careful assessment and advanced medical care, TOLAC is still a safe option for delivery. The objective of this study is to find the factors that impact the success rate for TOLAC and to compare the results with Taiwan national registry data. MATERIALS AND METHODS: A longitudinal cohort study that includes a total of 254 cases of women receiving TOLAC in a tertiary medical center over a period of 10 years. RESULTS: A total of 254 participants who underwent TOLAC, which accounts for 1.67% of total labor instances (254/15,166), were enrolled for analysis. The success rate of TOLAC was found to be 80.70% (205/254), including 146 (57.5%) normal deliveries, 45 (17.7%) vacuum-assisted deliveries, and 14 (5.5%) forceps-assisted deliveries. The conversion rate to cesarean section was 19.3%. There were no uterine rupture cases in our study, and there were only two suspected cases, which turned out to have no actual rupture. When analyzing the factors affecting the results of TOLAC, we found that a successfully spontaneously delivered baby had a lower birth weight than the failed TOLAC cases that were converted to cesarean delivery (mean, 2989 g vs. 3379 g; p < 0.001). Among the patients who were converted to cesarean section, the most common reason was dysfunctional labor (79.6%), followed by fetal distress (14.3%). CONCLUSION: Under intensive care and observation, TOLAC section may still be a feasible choice. Nevertheless, the body weight of the baby has been shown to be a factor that can influence the success rate.