Structural atlas of human primary microRNAs generated by SHAPE-MaP.

MicroRNA (miRNA) maturation is critically dependent on structural features of primary transcripts (pri-miRNAs). However, the scarcity of determined pri-miRNA structures has limited our understanding of miRNA maturation. Here, we employed selective 2'-hydroxyl acylation analyzed by primer extension and mutational profiling (SHAPE-MaP), a high-throughput RNA structure probing method, to unravel the secondary structures of 476 high-confidence human pri-miRNAs. Our SHAPE-based structures diverge substantially from those inferred solely from computation, particularly in the apical loop and basal segments, underlining the need for experimental data in RNA structure prediction. By comparing the structures with high-throughput processing data, we determined the optimal structural features of pri-miRNAs. The sequence determinants are influenced substantially by their structural contexts. Moreover, we identified an element termed the bulged GWG motif (bGWG) with a 3' bulge in the lower stem, which promotes processing. Our structure-function mapping better annotates the determinants of pri-miRNA processing and offers practical implications for designing small hairpin RNAs and predicting the impacts of miRNA mutations.

Robot-assisted total preservation of the pelvic autonomic nerve with extended systematic lymphadenectomy as part of nerve-sparing radical hysterectomy for cervical cancer.

OBJECTIVE: To evaluate our short-term clinical outcomes of robot-assisted autonomic nerve-sparing extended systematic lymphadenectomy as part of nerve-sparing radical hysterectomy. METHODS: Between March 2011 and June 2012, we observed prospectively 28 consecutive patients who underwent robot-assisted autonomic nerve-sparing extended systematic lymphadenectomy, including the superior and inferior gluteal, presacral (subaortic), common iliac, and lower para-aortic nodes. RESULTS: The predominant International Federation of Gynecology and Obstetrics stage was IB1 (15 patients), followed by IB2 (5 patients), IA2 (3 patients), IIA1 (3 patients), and IIA2 (2 patients). The mean ± SD total operating time was 308.8 ± 54.9 minutes, and the mean ± SD console time was 280.0 ± 46.0 minutes. The mean ± SD blood loss was 102.7 ± 153.8 mL. The mean ± SD acquired pelvic lymph node was 27.1 ± 9.3, the mean ± SD extended lymph node was 19.2 ± 9.6, and the mean ± SD total lymph node was 46.3 ± 14.5. A total of 10 patients (35.7%) had nodal metastasis; among them, 6 patients (21.4%) had single pelvic nodal metastasis, 3 patients (10.7%) had concurrent pelvic and extended nodal metastasis, and one patient (3.6%) had single extended nodal metastasis. No intraoperative complications that required treatment occurred; however, ureterovaginal fistula was identified in 4 patients (14.3%) and ureter stricture in 4 patients (14.3%) after radiotherapy. After a median follow-up of 10 months (range, 1-16 months), there was no pelvic recurrence; however, one patient had recurrence at transposition site of ovary. CONCLUSIONS: With the advantage of delicate movement of robot instrument, robot-assisted systematic extended lymphadenectomy with total preservation of pelvic autonomic nerves did not compromise the radicality, and its surgical technique was feasible and safe. By using this approach, we could harvest more lymph nodes and have a high rate of metastatic nodes without disturbing voiding function; however, there was increased rate of urological complications. Moreover, long-term survival benefit after an extended systematic lymphadenectomy must be evaluated.

Robot versus laparoscopic nerve-sparing radical hysterectomy for cervical cancer: a comparison of the intraoperative and perioperative results of a single surgeon's initial experience.

OBJECTIVE: The aim of the study was to compare the initial surgical outcomes and learning curve of nerve-sparing robotic radical hysterectomy (RRH) with nerve-sparing total laparoscopic radical hysterectomy (TLRH) for the treatment of early-stage cervical cancer in the first 50 cases. METHODS: Between January 2008 and March 2012, 50 consecutive patients underwent nerve-sparing RRH. These patients were compared with a historic cohort of the first 50 consecutive patients who underwent nerve-sparing TLRH. RESULTS: Both groups were similar with respect to patients and tumor characteristics. The mean operating time in the RRH group was significantly longer than that in the TLRH group (230.1 ± 35.8 vs 211.2 ± 46.7 minutes; P = 0.025). The mean blood loss for the robotic group was significantly lower compared with the laparoscopic group (54.9 ± 31.5 vs 201.9 ± 148.4 mL; P < 0.001). There was no significant difference in the mean pelvic lymph nodes between the 2 groups (25.0 ± 9.9 vs 23.1 ± 10.4; P = 0.361). The mean days to normal residual urine were 9.6 ± 6.4 in RRH and 11.0 ± 6.2 in TLRH (P = 0.291). The incidence of intraoperative complication was profoundly lower in RRH compared with that of TLRH (0% vs 8%; P = 0.041). Moreover, no intraoperative transfusion was required in RRH, whereas 4 (8%) were required in TLRH (P = 0.041). In both groups, we found no evidence of a learning effect during the first 50 cases. CONCLUSIONS: During the first 50 cases, surgical outcomes and complication rates of nerve-sparing RRH were found to be comparable to those of nerve-sparing TLRH. Moreover, the mean blood loss and intraoperative complication rate in the robotic group were significantly lower than those in the laparoscopic group. Surgical skills for nerve-sparing TLRH easily and safely translated to nerve-sparing RRH in case of experienced laparoscopic surgeon.

Multivalent nanofibers of a controlled length: regulation of bacterial cell agglutination.

Control of the size and shape of molecular assemblies on the nanometer scale in aqueous solutions is very important for the regulation of biological functions. Among the well-defined supramolecular structures of organic amphiphiles, one-dimensional nanofibers have attracted much attention because of their potential applications in biocompatible materials. Although much progress has been made in the field of self-assembled nanofibers, the ability to control the fiber length remains limited. The approach for control of the fiber length presented herein overcomes this limitation through the coassembly of amphiphilic rod-coil molecules in which the crystallinity of the aromatic segment can be regulated by π-π stacking interactions. The introduction of carbohydrate segments into the fiber exterior endows the nanofibers with the ability to adhere to bacterial cells. Notably, the fiber length systematically regulates the agglutination and proliferation of bacterial cells exposed to these fibers.

Multifunctional synthetic nano-chaperone for peptide folding and intracellular delivery.

Artificial, synthetic chaperones have attracted much attention in biomedical research due to their ability to control the folding of proteins and peptides. Here, we report bio-inspired multifunctional porous nanoparticles to modulate proper folding and intracellular delivery of therapeutic α-helical peptide. The Synthetic Nano-Chaperone for Peptide (SNCP) based on porous nanoparticles provides an internal hydrophobic environment which contributes in stabilizing secondary structure of encapsulated α-helical peptides due to the hydrophobic internal environments. In addition, SNCP with optimized inner surface modification not only improves thermal stability for α-helical peptide but also supports the peptide stapling methods in situ, serving as a nanoreactor. Then, SNCP subsequently delivers the stabilized therapeutic α-helical peptides into cancer cells, resulting in high therapeutic efficacy. SNCP improves cellular uptake and bioavailability of the anti-cancer peptide, so the cancer growth is effectively inhibited in vivo. These data indicate that the bio-inspired SNCP system combining nanoreactor and delivery carrier could provide a strategy to expedite the development of peptide therapeutics by overcoming existing drawbacks of α-helical peptides as drug candidates.

Progression rate from new-onset pre-hypertension to hypertension in Korean adults.

BACKGROUND: There are limited studies conducted in Asia to investigate the progression rate to hypertension (HTN). This study was done to estimate the progression rate of new-onset pre-HTN (PreHTN) to HTN during an 8-year follow-up period, and to compare the impact of PreHTN on progression to HTN. METHODS AND RESULTS: A total of 49,228 participants, aged 30 to 54 years with new-onset PreHTN at baseline (1994-1996) from a biennial national medical exam were enrolled and followed up every 2 years until 2004. The incidence rate recorded at each interval and the cumulative incidence rate of HTN were analyzed. Hazard ratio of high-normal and high blood pressure (BP) in men and women was calculated. The cumulative incidence rate for high-normal BP was 27.6% and 26.4% at 2-year follow-up, increased to respectively 64.1% and 55.8% in men and women at the 8-year follow-up. Compared to optimal BP, hazard ratios for men with high-normal BP across all age groups were 3- to 4-fold higher at 2-year, and 2- to 3-fold higher at 8-year follow-up. Hazard ratios for women were about 6-fold higher at 2-year and around 4-fold higher at 8-year follow-up. CONCLUSIONS: New PreHTN was a significant predisposing factor for future HTN, in young adults and the effect is more prominent in women.

Oncologic results and surgical morbidity of laparoscopic nerve-sparing radical hysterectomy in the treatment of FIGO stage IB cervical cancer: long-term follow-up.

OBJECTIVES: The aim of this study was to evaluate a long-term follow-up data for oncologic results and surgical morbidity of a laparoscopic nerve-sparing radical hysterectomy (NSRH) in the treatment of FIGO stage IB cervical cancer. METHODS: This was a retrospective study that comprised consecutive 125 patients with cervical cancer stage IB1 (n = 105) and IB2 (n = 20) who underwent a laparoscopic NSRH (Piver type III) by a gynecologic oncologist without selecting patients from January 1999 to December 2007. RESULTS: In regression analysis, the operating time (R linear = 0.311, P < 0.001) and estimated blood loss (R linear = 0.261, P < 0.001) were decreased, whereas the number of harvested pelvic lymph nodes (R linear = 0.250, P < 0.001) was increased. Seventeen patients (13.6%, 17/125) were found to have pelvic node metastasis. Para-aortic node metastasis had occurred in 2 patients (5.1%, 2/39). There were high urological complications (13/125, 10.4%) related to radical surgery. Forty-one patients (33%) needed transfusions. Positive surgical margins did not exist. Patients were able to self-void at a mean of 10.3 days postoperatively. The return rates to normal voiding function at postoperative 14 and 21 days were 92.0% and 95.2%, respectively. Thirteen patients (IB1 n = 9, IB2 n = 4) experienced a recurrence postoperatively. Six patients (IB1 n = 3, IB2 n = 3) died of recurrent disease. Five-year disease-free survival rates of cervical cancer IB1 and IB2 were 92% and 78%, respectively (P = 0.1772). Five-year overall survival rates of cervical cancer IB1 and IB2 were 96% and 83%, respectively (P = 0.0437). CONCLUSIONS: A laparoscopic NSRH for FIGO stage IB cervical cancer was comparable to open NSRH in terms of early recovery of bladder function. It did not compromise surgical radicality, but revealed high urological complications, long operating time, and much blood loss, compared with conventional radical hysterectomy. However, these surgical morbidities were corrected with increase in experiences.

Robotic uterine artery preservation and nerve-sparing radical trachelectomy with bilateral pelvic lymphadenectomy in early-stage cervical cancer.

OBJECTIVE: The aim of the study was to evaluate the safety and feasibility of robotic uterine artery preservation and nerve-sparing radical trachelectomy with pelvic lymphadenectomy using the da Vinci surgical system. METHODS: Three patients who were diagnosed with early-stage cervical cancer underwent robotic uterine artery preservation and nerve-sparing radical trachelectomy with bilateral lymphadenectomy from January 2010 to March 2010. The data were compared with those of 4 cases of total laparoscopic nerve-sparing radical trachelectomy that were performed from July 2004 to May 2005 and were previously reported. RESULTS: In the robotic group, the mean console time was 275 minutes (range, 240-305 minutes). The mean postoperative hemoglobin change was 0.4 g/dL (range, 0.2-0.6 g/dL). The mean estimated blood loss was 23 mL (range, 15-40 mL), which is less than that of the laparoscopic group. There were no metastases detected in any of the cases, and the resection margins were negative in both groups. CONCLUSIONS: The robotic uterine artery preservation and nerve-sparing radical trachelectomy with pelvic lymphadenectomy were efficient in reducing blood loss and feasible methods such as other approaches.

Prenatal diagnosis of scalp cyst with spontaneous regression.

Fetal scalp cysts are rare, and they may be easily misdiagnosed as meningoceles or encephaloceles. A 38-year-old pregnant woman was referred to our hospital for ultrasound examination, which revealed a small cyst above the skull with no defect in the skull. The cyst could not be seen on follow-up ultrasound examinations. After delivery, a three-dimensional CT scan of the head revealed no defect in skull and scalp. Fetal scalp cysts usually require surgical excision after birth, but can regress without any treatment.

A case of uterine pseudoaneurysm combined with gestational trophoblastic disease in a 30-year-old woman.

Uterine artery pseudoaneurysm is a rare disease and it can be diagnosed using conventional doppler ultrasongraphy. Damaged uterine arteries from cesarean section, myomectomy, dilatation & curettage, etc. are known as causes of the disease. Massive bleeding in the rupture can cause fatal result. We observed an increase in ß-hCG and uterine artery pseudoaneurysm a year after the performance of dilatation & curettage for hydatidiform mole and treated it with arterial embolization and chemotherapy. We report the case and give a brief review of the literature.

Laparoscopic second look operation for ovarian cancer: single center experiences.

We compared the clinical outcomes of laparoscopic surgery with those of laparotomy in second-look operations for ovarian cancer. We retrospectively reviewed the medical records of 35 patients treated between January 2000 and December 2005. They were categorized into two groups: Laparoscopy versus laparotomy group. Among 35 patients, 18 patients (51.4%) were treated with laparoscopy, and 17 patients (48.6%) were treated with laparotomy. There were no statistically significant differences except for hospital stay (laparoscopy vs. laparotomy five days vs. nine days, p < 0.05). Eight patients (44.4%) in the laparoscopy group and six (35.3%) in the laparotomy group were found to be pathologically positive after the second look operation. There were two recurrences (20.0%) in the laparoscopy group and four (36.4%) in the laparotomy group pathologically negative in the second-look operation (p = 0.557). There were no differences in disease-free survival rates (p = 0.705) between the two groups. The second look operation is an ideal method for histologic comfirmation of recurrent or persistent ovarian cancer. Laparoscopic surgery seems a safe and accurate method for the second-look examination, and can also reduce patient discomfort, hospital stay and morbidity associated with laparotomy.

Vaginal evisceration after total laparoscopic radical hysterectomy in cervical cancer.

A 50-year-old woman came to the emergency department with vaginal evisceration that occurred 7 months after a total laparoscopic radical hysterectomy. Vaginal evisceration was repaired by a laparoscopic-vaginal approach without a laparotomy. This is the first report of vaginal evisceration after a total laparoscopic radical hysterectomy.

The effect of nonperitonization and laparoscopic lymphadenectomy for minimizing the incidence of lymphocyst formation after radical hysterectomy for cervical cancer.

OBJECTIVES: To determine the clinical effect of nonperitonization and laparoscopic lymphadenectomy using bipolar electrocautery after radical hysterectomy for cervical cancer compared with peritonization and open lymphadenectomy using monopolar electrocautery. MATERIALS AND METHODS: This was a retrospective study of 180 patients who underwent a radical hysterectomy and pelvic lymph node dissection (PLND) for cervical cancer from August 1998 to August 2007. The patients were composed of the Peritonization + Laparotomy group (group A; n = 98, 196 PLND) and the Nonperitonization + Laparoscopy group (group B; n = 82, 164 PLND). Group B left the peritoneum in front of the PLND open. Two closed-suction drains were placed at each side of the PLND. RESULTS: Group B statistically showed a lesser total amount of lymphatic drainage (P = 0.001), shorter duration of inserted drains (P < 0.001), and shorter length of hospital stay (P < 0.001), compared with group A. The formation of lymphocysts occurred in 30 patients (41 lymphocysts) of group A and 5 patients (5 lymphocysts) of group B. Lymphocyst formation of group B had a statistically lower incidence than that of group A (P < 0.001). Lymphocyst formation was associated with an increase in the total amount of lymphatic drainage in group A (P = 0.090) and group B (P = 0.041) and a pathologic type of adenocarcinoma in group B (P = 0.016). Surgical experiences were not correlated with lymphocyst formation. CONCLUSIONS: The omission of peritonization and laparoscopic lymphadenectomy using bipolar electrocautery in early-stage cervical cancer were more effective than peritonization and open lymphadenectomy using monopolar electrocautery in minimizing the incidence of lymphocyst formation.

Immunohistochemical expression of YKL-40 in peritoneal endometriosis.

AIMS: To evaluate the relationship between the immunohistochemical expression of YKL-40 and peritoneal endometriosis by using paraffinized peritoneal tissue blocks. METHODS AND MATERIALS: We retrospectively collected data from 27 patients whose pathologic reports indicated invasion of the peritoneum by endometriosis. A conventional peroxidase staining technique was performed using rabbit polyclonal antibody (Quidel corporation, Santa Clara, CA) on peritoneal tissue blocks; their histology was then reviewed by a pathologist, and data were analyzed by nonparametric and Mann-Whitney tests. RESULTS: YKL-40 was detected immunohistochemically in 17 (63%) of 27 cases. The number of biopsies which were invaded by endometriosis (P = 0.015), with a score based on the revised classification of endometriosis (American Society for Reproductive Medicine) had statistical significance (P = 0.001). The obstetric history, age, body mass index, severity, and occurrence of dysmenorrhea, menstrual phase, preoperative CA 125, erythrocyte sedimentation rate, and white blood cell count had no statistical significance. YKL-40 immunoreactivity was recognized as brown staining, localized to the cytoplasm of epithelial cells of endometrial gland in peritoneal endometriosis. There was no positive staining on endometrial stromal cells or smooth muscle cells. CONCLUSIONS: YKL-40 is related to severity of peritoneal endometriosis. However why a different expression level of immunohistochemical staining should occur is not known and needs further investigation.

Clinical significance of serum albumin level in pregnancy-related hypertension.

AIM: To evaluate the usefulness of serum albumin level as a marker of severity in pregnancy-related hypertension. METHODS: Of 454 patients with pregnancy-related hypertension who were admitted to Kyungpook National University Hospital between May 1999 and April 2008, the medical records and laboratory tests of 354 patients who met the inclusion criteria for the current study were reviewed. A comparison of the characteristics of each hypertension group and the correlation between serum albumin levels and the time to delivery, 24-h urine protein, and/or pregnancy outcomes were statistically analyzed using SPSS 12.0 (SPSS Korea, Korea). RESULTS: Serum albumin level had a negative correlation with 24-h urine protein (Pearson's correlation coefficient = -0.481) and a positive correlation with time to delivery (= 0.389). Serum albumin ≤3.0 g/dL was highly associated with severe proteinuria (>2 g/day). There were significant differences in maternal or perinatal morbidity as a function of serum albumin level. If serum albumin level fell below 2.5 g/dL, the risks of ascites, hemolysis elevated liver enzyme low platelet (HELLP) syndrome and perinatal mortality significantly increased (odds ratio [OR] and 95% confidence interval [CI]: 3.5 [1.5-8.1], 12 [3.1-45], and 6.1 [1.7-22], respectively). CONCLUSION: Serum albumin level in pregnancy-related hypertension is a significant determinant of disease severity and may be considered as a useful marker for predicting time to delivery, severe proteinuria, and pregnancy outcomes.

Laparoscopic pelvic anatomy of nerve-sparing radical hysterectomy.

Many reports regarding nerve-sparing radical hysterectomy have been published. However, most reports have been based on systematic descriptions via laparotomy or cadaver dissection. The aim of this work was to describe the pelvic anatomy of nerve-sparing radical hysterectomy via laparoscopy, with specific focus on the inferior hypogastric plexus. This study is based on 125 patients with FIGO stage IB cervical cancer who had undergone laparoscopic nerve-sparing radical hysterectomies since 1999. The inferior hypogastric plexus was demonstrated via laparoscopy and was comprised of afferent fibers from the sacral root (S2, S3, and S4), sacral sympathetic ganglion, and hypogastric nerve, and efferent fibers forming its vesical, uterovaginal, and rectal branches. During the dissection of the posterior leaf of the vesicouterine ligament, various vesical veins were identified. If the cut edge of an inferior vesical vein was pulled medially with upward traction, the vesical branches of the inferior hypogastric plexus were exposed and these were divided into medial and lateral branches. The magnified view of laparoscopy made it possible to dissect nerves and vessels meticulously and to secure a clear resection margin during the dissection of the deep part of the cardinal ligament, uterosacral ligament, and posterior leaf of the vesicouterine ligament.

Learning curve of laparoscopic radical hysterectomy with pelvic and/or para-aortic lymphadenectomy in the early and locally advanced cervical cancer: comparison of the first 50 and second 50 cases.

BACKGROUND: To compare the surgical and oncological outcomes and morbidity of the first 50 cases treated by laparoscopic radical hysterectomy with those of the second 50 cases. METHODS: Between October 1994 and January 2004, we retrospectively reviewed the charts of 100 consecutive patients (International Federation of Gynecology and Obstetrics stages IA2 [n = 12], IB1 [n = 56], IB2 [n = 15], IIA [n = 15], and IIB [n = 2]) who underwent laparoscopic radical hysterectomy with pelvic and/or para-aortic lymphadenectomy. One hundred patients were divided into the first 50 cases (group 1) and second 50 cases (group 2). RESULTS: Operating time, length of hospital stay, time to normal residual urine, and transfusion rate significantly decreased, and the acquired number of pelvic nodes significantly increased when comparing group 1 with group 2. The intraoperative and postoperative complication rates profoundly decreased in group 2 as compared with group 1. After a median follow-up of 66.5 months, 10 patients had a recurrence, 9 of whom died. The 5-year overall survival rates were 96% in group 1 and 90% in group 2, and 5-year disease-free survival rates were 92% in group 1 and 90% in group 2. CONCLUSIONS: Laparoscopic radical hysterectomy is a feasible and safe treatment modality in early and even locally advanced cervical cancer without decreasing survival. Surgical outcome was improved with experience, and the complication rate related to operation of group 1 was higher than that of group 2. There was no significant difference in survival between the 2 groups.

Total laparoscopic nerve-sparing radical trachelectomy.

OBJECTIVES: To evaluate the feasibility and operative morbidity of a total laparoscopic nerve-sparing radical trachelectomy with pelvic lymphadenectomy. METHODS: Between July 2004 and May 2005, this procedure had been undergone to four consecutive young patients with early cervical cancer (one with stage IA2, three with stage IB1). It was distinguished from the conventional radical trachelectomy in that it consisted of a reanastomosis of uterine corpus and upper vagina via laparoscopic approach, and an autonomic nerve-sparing dissection under the magnified laparoscopic view. RESULTS: The mean operative time and blood loss was 250 minutes and 185 mL, respectively. The pathologic diagnosis was all squamous cell carcinoma. There were no node metastasis and cancer-free cervical margin. Any wound defect and leakage from a stump were not found. There were no neurologic impairments such as bladder hypotonia. The mean follow-up period was 34 months. Only one patient (Case 1) received concurrent chemo-radiation due to recurrence, followed by no further sign of progression. Unfortunately, there has not been any pregnancy yet. CONCLUSION: A total laparoscopic nerve-sparing radical trachelectomy is feasible and without any increase of morbidity, and has the potential to improve surgical outcomes, compared with a conventional radical trachelectomy.

High-throughput chemical screening to discover new modulators of microRNA expression in living cells by using graphene-based biosensor.

MicroRNAs (miRNAs) are important regulatory RNAs that control gene expression in various biological processes. Therefore, control over the disease-related miRNA expression is important both for basic research and for a new class of therapeutic modality to treat serious diseases such as cancer. Here, we present a high-throughput screening strategy to identify small molecules that modulate miRNA expression in living cells. The screen enables simultaneous monitoring of the phenotypic cellular changes associated with the miRNA expression by measuring quantitative fluorescent signals corresponding to target miRNA level in living cells based on a novel biosensor composed of peptide nucleic acid and nano-sized graphene oxide. In this study, the biosensor based cellular screening of 967 compounds (including FDA-approved drugs, enzyme inhibitors, agonists, and antagonists) in cells identified four different classes of small molecules consisting of (i) 70 compounds that suppress both miRNA-21 (miR-21) expression and cell proliferation, (ii) 65 compounds that enhance miR-21 expression and reduce cell proliferation, (iii) 2 compounds that suppress miR-21 expression and increase cell proliferation, and (iv) 21 compounds that enhance both miR-21 expression and cell proliferation. We further investigated the hit compounds to correlate cell morphology changes and cell migration ability with decreased expression of miR-21.

A robust and quantitative assay platform for multiplexed, high throughput screening of protein kinase inhibitors.

We present a new platform for multiplexed protein kinase activity assay using TiO2 decorated graphene oxide (GO), which is applicable to high throughput inhibitor screening. On the basis of the strong affinity of TiO2 for the phosphate group and the fluorescence quenching capability of GO, phosphorylation of substrates by protein kinases was quantitatively measured in a short time.