Chien-Nan Lee

Interleukin-6 promotes cervical tumor growth by VEGF-dependent angiogenesis via a STAT3 pathway.

Interleukin-6 (IL-6) has received particular attention in the pathogenesis of cervical cancer, although the underlying mechanism remains elusive. This study revealed that IL-6 promotes in vivo tumor growth of human cervical cancer C33A cells, but does not substantially alter their in vitro growth kinetics. The in vivo angiogenic assays showed that IL-6 increases angiogenic activity in human cervical cancer cells, an effect that is specifically associated with upregulation of vascular endothelial growth factor (VEGF). Also, using anti-VEGF antibody to block VEGF function significantly inhibited IL-6-mediated angiogenesis and tumor growth in nude mice, strongly supporting the critical role of VEGF in the IL-6-mediated cervical tumorigenesis. Accordingly, the signaling pathway downstream of IL-6/IL-6R responsible for the regulation of VEGF was investigated. Notably, pharmacological inhibition of PI3-K or MAPK failed to inhibit IL-6-mediated transcriptional upregulation of VEGF. Meanwhile, blocking STAT3 pathway with dominant-negative mutant STAT3D effectively abolished IL-6-induced VEGF mRNA. In transient transfections, a luciferase reporter construct containing the full-length 1.5-kb VEGF promoter or a 1.2-kb fragment lacking the known hypoxic-response element also exhibited the same degree of response to IL-6. Additionally, transient transfection of STAT3D downregulated the 1.2-kb VEGF promoter luciferase reporter stimulated by IL-6. Based on the above phenomenon combined with the concomitant increased tumor expression of IL-6 and VEGF in cervical cancer tissues, we conclude that IL-6 may promote cervical tumorigenesis by activating VEGF-mediated angiogenesis via a STAT3 pathway.

Role of three‐dimensional power Doppler in the antenatal diagnosis of placenta accreta: comparison with gray‐scale and color Doppler techniques

To assess the role of three‐dimensional (3D) power Doppler in the antenatal diagnosis of placenta accreta and compare its diagnostic performance with gray‐scale and color Doppler ultrasonography.

Clinical utility of array comparative genomic hybridisation for prenatal diagnosis: a cohort study of 3171 pregnancies

Sir, We read with great interest your recent publication by Lee et al. on the use of array comparative genomic hybridisation (CGH) for prenatal diagnosis: a cohort study of 3171 pregnancies. Such a large cohort is an important contribution to the existing literature. It is important then to compare and contrast the results of Lee’s study with those of other recently published cohorts. The abstract states that when array CGH is performed for a fetus with a congenital anomaly apparent on prenatal ultrasound, the test was effective in identifying submicroscopic genomic imbalances (i.e. that are unidentifiable by conventional G-band karyotyping) in 33 of 194 (17%) cases. However, on reviewing data within table 2 (and later on in the abstract) this rate is recorded as 16/194 (8.2%). We attempted to extract data on submicroscopic copy number changes from table 2, but there appeared to have been formatting errors (e.g. in the row ‘pathogenic karyotyping reports’ the columns appear to be shifted a space to the right). It is unclear within the table how the calculated total of pathogenic karyotype and array CGH reports are constructed. We assume that pathogenic karyotype reports would be made up of those with numerical abnormality plus those with large chromosomal deletions and duplications, whereas pathogenic array reports would be made up of the former plus microscopic chromosomal deletions and duplications, plus variants of unknown significance. This, however, is not made clear. It is not certain whether there may have been instances where the array CGH test failed to identify a chromosomal anomaly detected by conventional karyotyping, and indeed whether both types of karyotype testing were performed for all fetuses included in the study. The authors state within the discussion that ‘when array CGH was performed alone no significantly pathological results were missed’, but without a reference test how could one be sure? Two array CGH platforms were used within this study: a 1-Mb bacterial artificial chromosome (BAC) and a 60-kb oligonucleotide array. It appears that there was a switch to the higher resolution 60-kb array towards the end of the cohort study. However, table 3 is not transparent to which array was used to detect which abnormal result. When extrapolating using the coordinates of the oligonucleotide probes in comparison with those of BAC probes aligned to the reference genome in ensembl, and in BlueGnome’s bluefuse multi database software, we predict that only one of the abnormal results detected by the higher resolution 60-kb array within Lee et al.’s cohort would have been missed by the 1-Mb BAC. This point should be commented on, as the lower resolution BAC would be predicted to produce less variants of unknown significance (VOUS), an important point to consider when using this technology on prenatal samples. The authors did not however state which array contributed to the five VOUS recorded. Finally, the authors conclude that there is a 0.52% baseline risk of submicroscopic genomic imbalance, even in women with an uneventful prenatal ultrasound examination. The authors conclude that prenatal array CGH is a recommended additional test in pregnant women undergoing invasive testing, but do not comment on the additional VOUS results that such testing could reveal, and the problems of interpretation and counselling of these variants in the prenatal setting. Before we can think about using this technology, even as an adjunct to conventional karyotypic testing, this issue must be addressed and the additional costs to overall pregnancy health care must be considered. j

Efficacy of maternal tenofovir disoproxil fumarate in interrupting mother-to-infant transmission of hepatitis B virus

The efficacy and safety of maternal tenofovir disoproxil fumarate (TDF) in reducing mother‐to‐infant hepatitis B virus (HBV) transmissions is not clearly understood. We conducted a prospective, multicenter trial and enrolled 118 hepatitis B surface antigen (HBsAg)– and hepatitis B e antigen–positive pregnant women with HBV DNA ≥7.5 log10 IU/mL. The mothers received no medication (control group, n = 56, HBV DNA 8.22 ± 0.39 log10 IU/mL) or TDF 300 mg daily (TDF group, n = 62, HBV DNA 8.18 ± 0.47 log10 IU/mL) from 30‐32 weeks of gestation until 1 month postpartum. Primary outcome was infant HBsAg at 6 months old. At delivery, the TDF group had lower maternal HBV DNA levels (4.29 ± 0.93 versus 8.10 ± 0.56 log10 IU/mL, P < 0.0001). Of the 121/123 newborns, the TDF group had lower rates of HBV DNA positivity at birth (6.15% versus 31.48%, P = 0.0003) and HBsAg positivity at 6 months old (1.54% versus 10.71%, P = 0.0481). Multivariate analysis revealed that the TDF group had lower risk (odds ratio = 0.10, P = 0.0434) and amniocentesis was associated with higher risk (odds ratio 6.82, P = 0.0220) of infant HBsAg positivity. The TDF group had less incidence of maternal alanine aminotransferase (ALT) levels above two times the upper limit of normal for ≥3 months (3.23% versus 14.29%, P = 0.0455), a lesser extent of postpartum elevations of ALT (P = 0.007), and a lower rate of ALT over five times the upper limit of normal (1.64% versus 14.29%, P = 0.0135) at 2 months postpartum. Maternal creatinine and creatinine kinase levels, rates of congenital anomaly, premature birth, and growth parameters in infants were comparable in both groups. At 12 months, one TDF‐group child newly developed HBsAg positivity, presumably due to postnatal infection and inefficient humoral responses to vaccines. Conclusions: Treatment with TDF for highly viremic mothers decreased infant HBV DNA at birth and infant HBsAg positivity at 6 months and ameliorated maternal ALT elevations. (Hepatology 2015;62:375–386

Vascular endothelial growth factor and prognosis of cervical carcinoma

Objective To evaluate vascular endothelial growth factor (VEGF) as a marker for predicting lymph node metastasis and an independent prognostic factor of early-stage cervical carcinoma. Methods One hundred thirty-five women with stage IB–IIA cervical carcinoma had radical abdominal hysterectomies and pelvic lymph node dissections. Intratumoral cytosol VEGF concentrations were assayed with enzyme immunoassay. Histopathologic items and cytosol VEGF-influencing clinical outcomes were compared. Results Twenty-two women (16.3%) who had disease recurrence had higher levels of cytosol VEGF (1020 versus 112 pg/mg protein, P < .001) than those without recurrence. Using a cutoff value of 400 pg/mg protein resulted in best sensitivity of 75%, best specificity of 70%, positive predictive value of 41%, and negative predictive value of 92%. Only overexpressed cytosol VEGF (hazard ratio 6.44, P < .001) was an independent prognostic factor of disease-free survival. The overexpressed cytosol VEGF (hazard ratio 4.50, P = .021) and positive lymphovascular emboli (hazard ratio 4.11, P = .045) were independent prognostic factor of overall survival. Conclusion Cytosol VEGF might be a biomarker for the status of pelvic lymph nodes in early-stage cervical carcinoma and an independent prognostic indicator of its outcome.

A whole genome methylation analysis of systemic lupus erythematosus: hypomethylation of the IL10 and IL1R2 promoters is associated with disease activity

The etiology of systemic lupus erythematosus (SLE) involves a complex interaction of genetic and environmental factors. Investigations have shown that environmentally driven epigenetic changes contribute to the etiology of SLE. Here, we hypothesize that aberrant DNA methylation may contribute to the activation of the immune machinery and trigger lupus disease activity. A whole genome methylation array was applied to investigate the DNA methylation changes between 12 pairs of active SLE patients and healthy controls. The results were further confirmed in 66 SLE patients, 102 healthy controls. The methylation statuses of the IL10 and IL1R2 genes were significantly reduced in the SLE patient samples relative to the healthy controls (age-adjusted odds ratios, 64.2 and 16.9, respectively, P<0.0001). There was a trend toward SLE patients having hypomethylated IL10 and IL1R2 genes accompanied by greater disease activity. We observed that the methylation degree of IL10 and IL1R2 genes were reduced in the rheumatoid arthritis (RA) patients as well but the hypomethylation change was more significant in IL1R2 genes than in the IL10 genes in RA patients. This study demonstrated that DNA hypomethylation might be associated with SLE. Hypomethylated IL10 and IL1R2 genes may provide potential epigenetic markers as clinical predictors for autoimmune diseases.

Antenatal Depiction of the Fetal Ear With Three-Dimensional Ultrasonography

Objective To evaluate the feasibility of examining the fetal ear with three-dimensional ultrasound. Methods In 125 pregnancies between 19 and 38 weeks of gestation, fetal ears were evaluated by three-dimensional ultrasound. The volume images with surface rendering were analyzed to depict the morphology, lying axis, orientation, and cranial location of the fetal ears. Results Three-dimensional images of one or both ears were successfully reconstructed in 105 fetuses. Among them, 18 fetuses had anomalous ears. The anomalous ears, including microtia, low-set ear with slope axis, abnormal ear orientation, and edematous ear, were confirmed after delivery. Three-dimensional ultrasound consistently displayed fetal ear abnormalities with greater accuracy and clarity. Conclusion Because anomalous ears may be a part of complex fetal malformations, it is important to recognize ear abnormalities. Due to the complexity of the fetal ear, three-dimensional ultrasound offers more important information than two-dimensional ultrasound, which simply gives auricular geometry. We suggest that three-dimensional ultrasound can be used better to examine the fetal ear and may prove to be useful for prenatal diagnosis and genetic counseling.

Molecular and clinical analyses of 84 patients with tuberous sclerosis complex

BackgroundTuberous sclerosis complex (TSC) is an autosomal dominant disease characterized by the development of multiple hamartomas in many internal organs. Mutations in either one of 2 genes, TSC1 and TSC2, have been attributed to the development of TSC. More than two-thirds of TSC patients are sporadic cases, and a wide variety of mutations in the coding region of the TSC1 and TSC2 genes have been reported.MethodsMutational analysis of TSC1 and TSC2 genes was performed in 84 Taiwanese TSC families using denaturing high-performance liquid chromatography (DHPLC) and direct sequencing.ResultsMutations were identified in a total of 64 (76 %) cases, including 9 TSC1 mutations (7 sporadic and 2 familial cases) and 55 TSC2 mutations (47 sporadic and 8 familial cases). Thirty-one of the 64 mutations found have not been described previously. The phenotype association is consistent with findings from other large studies, showing that disease resulting from mutations to TSC1 is less severe than disease due to TSC2 mutation.ConclusionThis study provides a representative picture of the distribution of mutations of the TSC1 and TSC2 genes in clinically ascertained TSC cases in the Taiwanese population. Although nearly half of the mutations identified were novel, the kinds and distribution of mutation were not different in this population compared to that seen in larger European and American studies.

Correlation of Prenatal Ultrasound and Postnatal Outcome in Meconium Peritonitis

Objectives: To study the relationship between prenatal ultrasound features and postnatal course of meconium peritonitis. Methods: Meconium peritonitis was diagnosed by prenatal ultrasound. Fetuses were treated by intrauterine paracentesis of ascites when indicated, and symptomatic newborns received surgery. Results: Totally 17 cases were enrolled. Prenatal ultrasound findings include abdominal calcification (16/17), fetal ascites (12/17), hydramnios (9/17), pseudocyst (7/17) and dilated bowel loop (6/17). Persistent ascites, pseudocyst or dilated bowel loop are most sensitive (92%) to predict postnatal surgery (p = 0.022). The survivors have a higher gestational age at birth (36.4 vs. 33.3 weeks, p = 0.008). Persistent ascites and postnatal persistent pulmonary hypertension of the newborns significantly correlate with neonatal mortality (p = 0.029 and 0.022). Conclusion: Prenatal ultrasound can predict the neonatal outcome in meconium peritonitis.

Fusion Protein Vaccine by Domains of Bacterial Exotoxin Linked with a Tumor Antigen Generates Potent Immunologic Responses and Antitumor Effects

Antigen-specific immunotherapy represents an attractive approach for cancer treatment because of the capacity to eradicate systemic tumors at multiple sites in the body while retaining the requisite specificity to discriminate between neoplastic and nonneoplastic cells. It has been shown that certain domains of bacterial exotoxins facilitate translocation from extracellular and vesicular compartments into the cytoplasm. This feature provides an opportunity to enhance class I and/or II presentation of exogenous antigen to T lymphocytes. We investigated previously whether the translocation domain (domain II) of Pseudomonas aeruginosa exotoxin A with a model tumor antigen, human papillomavirus type 16 E7, in the context of a DNA vaccine could enhance vaccine potency. We then attempted to determine whether this chimeric molecule could also generate strong antigen-specific immunologic responses and enhance the potency of cancer vaccine in the protein format. Our results show that vaccination with the PE(DeltaIII)-E7-KDEL3 fusion protein enhances MHC class I and II presentation of E7, leading to dramatic increases in the number of E7-specific CD8+ and CD4+ T-cell precursors and markedly raised titers of E7-specific antibodies. Furthermore, the PE(DeltaIII)-E7-KDEL3 protein generates potent antitumor effects against s.c. E7-expressing tumors and preestablished E7-expressing metastatic lung tumors. Further, mice immunized with PE(DeltaIII)-E7-KDEL3 protein vaccine also retained long-term immunologic responses and antitumor effects. Our results indicate that retrograde-fusion protein via the delivery domains of exotoxins with an antigen greatly enhances in vivo antigen-specific immunologic responses and represents a novel strategy to improve cancer vaccine potency.