Implication of androgen receptor gene dysfunction in human Müllerian duct anomalies.

BACKGROUND: Müllerian duct anomalies (MDAs) are congenital developmental disorders exhibiting as a variety of malformations of female reproductive tract. The identified etiology of MDAs is limited. The present study aimed to unravel the underlying genetic causes of MDAs. METHODS: Rare variants in androgen receptor (AR) were called from the cohort consists of patients with MDAs and underwent whole exome sequencing (WES) at Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China. Sanger sequencing was used to confirm the causative genetic mutations. In silico analysis were used to classify the pathogenicity of each variant. Molecular modeling and simulations were conducted to investigate the conformational changes between the wild-type (WT) and mutant proteins. RESULTS: A total of 3 rare heterozygous variants in AR from the MDAs cohort in our institution were identified, with unknown effects. All variants were missense mutations, including c.173A > T, c.558C > A and c.1208C > T, and were absent or rare in East Asian populations in Genome Aggregation Database and the Exome Aggregation Consortium Database. According to the American College of Medical Genetics and Genomics guidelines, c.1208C > T variant was classified as likely pathogenic, while the other two were variants of uncertain significance. During molecular dynamics simulations, WT and mutant proteins all reached stable status according to root-mean-square variance. Values of radius of gyration showed that Q58L and S186R protein would be more compact than WT, while the structure of A403V became looser. Despite, in comparison with WT, the number of hydrogen bonds increased in Q58L, while decreased in the other two variants. Furthermore, the solvent-accessible surface area diminished in Q58L and A403V while enlarged in S186R proteins, when compared with WT. CONCLUSIONS: To our knowledge, this is the first report regarding the association of AR mutation and MDAs. The identification of these variants, predicted to damage the structure and function of AR protein, not only expanded the mutational spectrum of causative genes of MDAs, but provide novel molecular genetic reference for future studies.

Variants in genes related to development of the urinary system are associated with Mayer-Rokitansky-Küster-Hauser syndrome.

Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome, also known as Müllerian agenesis, is characterized by uterovaginal aplasia in an otherwise phenotypically normal female with a normal 46,XX karyotype. Previous studies have associated sequence variants of PAX8, TBX6, GEN1, WNT4, WNT9B, BMP4, BMP7, HOXA10, EMX2, LHX1, GREB1L, LAMC1, and other genes with MRKH syndrome. The purpose of this study was to identify the novel genetic causes of MRKH syndrome. Ten patients with MRKH syndrome were recruited at Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China. Whole-exome sequencing was performed for each patient. Sanger sequencing confirmed the potential causative genetic variants in each patient. In silico analysis and American College of Medical Genetics and Genomics (ACMG) guidelines helped to classify the pathogenicity of each variant. The Robetta online protein structure prediction tool determined whether the variants affected protein structures. Eleven variants were identified in 90% (9/10) of the patients and were considered a molecular genetic diagnosis of MRKH syndrome. These 11 variants were related to nine genes: TBC1D1, KMT2D, HOXD3, DLG5, GLI3, HIRA, GATA3, LIFR, and CLIP1. Sequence variants of TBC1D1 were found in two unrelated patients. All variants were heterozygous. These changes included one frameshift variant, one stop-codon variant, and nine missense variants. All identified variants were absent or rare in gnomAD East Asian populations. Two of the 11 variants (18.2%) were classified as pathogenic according to the ACMG guidelines, and the remaining nine (81.8%) were classified as variants of uncertain significance. Robetta online protein structure prediction analysis suggested that missense variants in TBC1D1 (p.E357Q), HOXD3 (p.P192R), and GLI3 (p.L299V) proteins caused significant structural changes compared to those in wild-type proteins, which in turn may lead to changes in protein function. This study identified many novel genes, especially TBC1D1, related to the pathogenesis of MRKH syndrome. The identification of these variants provides new insights into the etiology of MRKH syndrome and a new molecular genetic reference for the development of the reproductive tract.

Renal agenesis-related genes are associated with Herlyn-Werner-Wunderlich syndrome.

OBJECTIVE: To explore the genetic causes of Herlyn-Werner-Wunderlich syndrome (HWWS) using whole-exome sequencing. DESIGN: Retrospective genetic study. SETTING: Academic medical center. PATIENT(S): Twelve patients with HWWS. INTERVENTION(S): Whole-exome sequencing was performed for each patient. Sanger sequencing was used to confirm the potential causative genetic variants. In silico analysis and American College of Medical Genetics and Genomics guidelines were used to classify the pathogenicity of each variant. MAIN OUTCOME MEASURE(S): Rare sequence variants associated with müllerian duct development and renal agenesis were identified and included in subsequent analyses. RESULT(S): A total of 11 variants were identified in 10 of 12 patients (83.3%) and were considered to constitute a molecular genetic diagnosis of HWWS. These 11 variants were related to 9 genes: CHD1L, TRIM32, TGFBR3, WNT4, RET, FRAS1, FAT1, FOXF1, and PCSK5. All variants were heterozygous and confirmed by Sanger sequencing. The changes included one frameshift variant, one splice-site variant, and eight missense variants. All of the identified variants were absent or rare in Genome Aggregation Database East Asian populations. One of the 11 variants (9.1%) was classified as a pathogenic variant according to the American College of Medical Genetics and Genomics guidelines, and 8 of the 11 variants (72.7%) were classified as variants of uncertain significance. CONCLUSION(S): To our knowledge, this is the first report of the genetic causes of HWWS. Renal agenesis-related genes, such as CHD1L, TRIM32, RET, and WNT4, may be associated with HWWS. Identification of these variants can not only help us understand the etiology of HWWS and the relationship between reproductive tract development and urinary system development, but additionally improve the level of genetic counseling for HWWS.

Whole-Exome Sequencing Identified a TBX6 Loss of Function Mutation in a Patient with Distal Vaginal Atresia.

STUDY OBJECTIVE: The purpose of this study was to determine if there are any genetic changes with whole-exome sequencing associated with distal vaginal atresia. DESIGN: This was a retrospective genetics study of 5 patients who presented with distal vaginal atresia who were recruited between 2017 and 2018. Whole-exome sequencing was performed in each subject with distal vaginal atresia. Sanger sequencing was used to confirm the potential causative genetic mutation. SETTING: Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China. PARTICIPANTS AND MAIN OUTCOME MEASURES: The main outcome measure was the rare mutations potentially associated with distal vaginal atresia in 5 patients. RESULTS: A truncating mutation c.266delC (p.P89Rfs*5) in the T-box transcription factor 6 (TBX6) gene, which is highly expressed in the human vagina, was identified in 1 patient using whole-exome sequencing. The deletion of the 16p11.2 region containing the TBX6 locus has also been reported previously to have the clinical feature of Müllerian agenesis. This mutation was paternally inherited by the patient. This truncating mutation was absent from all of the databases we checked, suggesting that the variant is rare and pathogenic. CONCLUSION: We showed, to our knowledge, for the first time, that the mutation in TBX6 might be associated with human distal vaginal atresia.

An Enantioselective Potentiometric Sensor for 2-Amino-1-Butanol Based on Chiral Porous Organic Cage CC3-R.

Porous organic cages (POCs) have attracted extensive attention due to their unique structures and tremendous application potential in numerous areas. In this study, an enantioselective potentiometric sensor composed of a polyvinyl chloride (PVC) membrane electrode modified with CC3-R POC material was used for the recognition of enantiomers of 2-amino-1-butanol. After optimisation, the developed sensor exhibited enantioselectivity toward S-2-amino-1-butanol ( log K S , R P o t = -0.98) with acceptable sensitivity, and a near-Nernstian response of 25.8 ± 0.3 mV/decade within a pH range of 6.0⁻9.0.

Intraoperative ultrasound-assisted enucleation of residual fibroids following laparoscopic myomectomy.

AIM: To investigate if intraoperative ultrasounds by laparoscopic and transvaginal ultrasonography (LUS and TVS) could improve enucleating the residual fibroids following laparoscopic myomectomy (LM). METHODS: From March to December 2016, 78 women with uterine fibroids underwent LM, LUS and TVS were applied to detect residual fibroids and to guide surgeons to enucleate them after the visible fibroids were removed during LM operation. RESULTS: The total number of residual fibroids found by LUS was 140, and the total number found by TVS was 127 following LM (P = 0.03). LUS is statistically superior to TVS in the detection of residual fibroids in the anterior wall (P = 0.004), in the detection of intramural fibroids (P = 0.002), and in the detection of fibroids with a diameter ranging from 0.5 to 1 cm (P = 0.002). According to the total number of enucleated fibroids by LM, patients were divided into three groups (Group 1: 2 to 4, Group 2: 5 to 7 and Group 3: ≥8 fibroid counts). The percentages of patients in each group with residual fibroids at the end of surgery were 22.2%, 51.9% and 66.7% respectively. CONCLUSIONS: Both LUS and TVS are beneficial to surgical treatment of fibroids by assisting enucleation of residual fibroids following LM, while LUS is more effective in localizing residual fibroids than TVS.

A chiral, porous, organic cage-based, enantioselective potentiometric sensor for 2-aminobutanol.

A new enantioselective potentiometric sensor containing R-type chiral porous organic cage CC9 as the chiral selector was designed for the assay of 2-aminobutanol. Optimized membrane electrodes displayed a linear dynamic range from 10-3  ~ 10-1  mol·L-1 with a detection limit of 2.5 × 10-4  mol·L-1 and a Nernstian response of 27 ± 0.5mV·decade-1 toward S-2-aminobutanol within the pH range 7.0-10.0. The potentiometric enantioselectivity coefficient ( logKS,RPot) of this sensor was -1.333, indicating that the porous organic cage-based electrode exhibited good discrimination toward S-2-aminobutanol over R-2-aminobutanol.

Optical resolution and mechanism using enantioselective cellulose, sodium alginate and hydroxypropyl-ß-cyclodextrin membranes.

Chiral solid membranes of cellulose, sodium alginate, and hydroxypropyl-ß-cyclodextrin were prepared for chiral dialysis separations. After optimizing the membrane material concentrations, the membrane preparation conditions and the feed concentrations, enantiomeric excesses of 89.1%, 42.6%, and 59.1% were obtained for mandelic acid on the cellulose membrane, p-hydroxy phenylglycine on the sodium alginate membrane, and p-hydroxy phenylglycine on the hydroxypropyl-ß-cyclodextrin membrane, respectively. To study the optical resolution mechanism, chiral discrimination by membrane adsorption, solid phase extraction, membrane chromatography, high-pressure liquid chromatography ultrafiltration were performed. All of the experimental results showed that the first adsorbed enantiomer was not the enantiomer that first permeated the membrane. The crystal structures of mandelic acid and p-hydroxy phenylglycine are the racematic compounds. We suggest that the chiral separation mechanism of the solid membrane is "adsorption - association - diffusion," which is able to explain the optical resolution of the enantioselective membrane. This is also the first report in which solid membranes of sodium alginate and hydroxypropyl-ß-cyclodextrin were used in the chiral separation of p-hydroxy phenylglycine.

Homochiral metal-organic framework used as a stationary phase for high-performance liquid chromatography.

Metal-organic frameworks are promising porous materials. Chiral metal-organic frameworks have attracted considerable attention in controlling enantioselectivity. In this study, a homochiral metal-organic framework [Co(2) (D-cam)(2) (TMDPy)] (D-cam = D-camphorates, TMDPy = 4,4'-trimethylenedipyridine) with a non-interpenetrating primitive cubic net has been used as a chiral stationary phase in high-performance liquid chromatography. It has allowed the successful separation of six positional isomers and six chiral compounds. The good selectivity and baseline separation, or at least 60% valley separation, confirmed its excellent molecular recognition characteristics. The relative standard deviations for the retention time of run-to-run and column-to-column were less than 1.8 and 3.1%, respectively. These results demonstrate that [Co(2) (D-cam)(2) (TMDPy)] may represent a promising chiral stationary phase for use in high-performance liquid chromatography.

Single-walled carbon nanotubes for improved enantioseparations on a chiral ionic liquid stationary phase in GC.

In order to investigate whether the use of single-walled carbon nanotubes can improve enantioseparations on an ionic liquid stationary phase, a chiral ionic liquid, (R)-N,N,N-trimethyl-2-aminobutanol-bis(trifluoromethanesulfon)imidate, was synthesized. Two capillary columns, one containing the chiral ionic liquid and the other containing the single-walled carbon nanotubes and the chiral ionic liquid, were then prepared for GC. The results of the separations achieved with these columns show that coating the chiral ionic liquid stationary phase onto the capillary column containing single-walled carbon nanotubes improves the enantioselectivety of the chiral ionic liquid. This work indicates that using single-walled carbon nanotubes in this manner enables the application range of such GC chiral separations to be extended.

[Effects of Spasfon on course of labor].

OBJECTIVE: To observe the effects of Spasfon on improving dilatation of cervix and promoting the progression of labor. METHODS: Ninety seven normal primiparae with cervical edema were randomly divided into Spasfon group (group A, n = 46) and atropine group (group B, n = 51) when the cervix dilated 2 - 3 cm. Group A was given 80 mg of Spasfon intravenously, and group B was injected atropine 0.5 mg into the cervix. RESULTS: (1) The mean time period from drug administration to full dilation of the cervix was (3.1 +/- 0.3) h in group A, and (4.4 +/- 0.4) h in group B (P < 0.01). (2) The disappearance ratio of cervical edema 2 h after drug administration in group A was 95.6%, while in group B it was 90.2% (P > 0.05); the mean dilatation of cervix between the 2 hours in group A was (4.3 +/- 0.2) cm, while in group B it was (2.5 +/- 0.3) cm (P < 0.01). (3) There were no obvious side effects in group A. While eight women in group B complained of thirst and 22 women had increased heart rate accompanied with elevated baseline FHR, which all recovered in about 60 minutes. (4) Vaginal delivery rate in group A was 95.7%, and 90.2% in group B (P > 0.05). (5) There was no statistically significant difference in the color of amniotic fluid, suffocation state and weight of the newborns between the two groups (P > 0.05). (6) There was no statistically significant difference in postpartum hemorrhage between the two groups, either (P > 0.05). CONCLUSION: Spasfon can effectively improve cervical dilatation during labor and it is well tolerated by both mother and newborn.