Anti-miR-141-3p maintains homeostasis between autophagy and apoptosis by targeting Yy1 in the fetal lumbosacral defecation center of rats.

Anorectal malformations (ARMs) are congenital diseases that lead to postoperative fecal incontinence, constipation, and soiling, despite improvements in surgery; however, their pathological mechanisms remain unclear. Here, we report the role of microRNA-141-3p in maintaining homeostasis between apoptosis and autophagy in the lumbosacral defecation center of fetal rats with ARMs. Elevated microRNA-141-3p expression inhibited YIN-YANG-1 expression by binding its 3' UTR, and repressed autophagy and triggered apoptosis simultaneously. Then, adenylate cyclase 3 was screened to be the downstream target gene of YIN-YANG-1 by chromatin immunoprecipitation sequencing experiments, and Yin Yang 1 could positively activate the transcription of adenylate cyclase 3 by directly interacting with the motif GAGATGG and ATGG in its promoter. Intraamniotic microinjection of adeno-rno-microRNA-141-3p-sponge-GFP in fetal rats with ARMs on embryonic day 15 restored apoptosis-autophagy homeostasis. These findings reveal that microRNA-141-3p upregulation impaired homeostasis between apoptosis and autophagy by inhibiting the YIN-YANG-1/adenylate cyclase 3 axis, and that intraamniotic injection of anti-microRNA-141-3p helped maintain homeostasis in the lumbosacral defecation center of ARMs during embryogenesis.

Therapeutic potential of adenovirus-encoding brain-derived neurotrophic factor for spina bifida aperta by intra-amniotic delivery in a rat model.

Spina bifida aperta is a type of neural tube defect (NTD). Although prenatal fetal surgery has been an available and effective treatment for it, the neurological functional recovery is still need to be enhanced. Our previous results revealed that deficiencies of sensory, motor, and parasympathetic neurons were primary anomalies that occurred with the spinal malformation. Therefore, we emphasized that nerve regeneration is critical for NTD therapy. We delivered an adenoviral construct containing genes inserted for green fluorescent protein and brain-derived neurotrophic factor (Ad-GFP-BDNF) into the amniotic fluid to investigate its prenatal therapeutic potential for rat fetuses with spina bifida aperta. Using immunofluorescence, TdT-mediated dUTP nick-end labeling staining, and real-time polymerase chain reaction analysis, we assessed cell apoptosis in the defective spinal cord and Brn3a positive neuron survival in the dorsal root ganglion (DRG); a protein array was used to investigate the microenvironmental changes of the amniotic fluid. We found that most of the overexpressed BDNF was present on the lesions of the spina bifida fetuses, the number of apoptosis cells in Ad-GFP-BDNF-transfected spinal cords were reduced, mRNA levels of Bcl2/Bax were upregulated and Casp3 were downregulated compared with the controls, the proportion of Brn3a positive neurons in DRG were increased by activating the BDNF/TrkB/Akt signaling pathway, and most of the significant changes in cytokines in the amniotic fluid were related to the biological processes of regulation of apoptotic process and generation of neurons. These results suggest that intra-amniotic Ad-GFP-BDNF gene delivery might have potential as a supplementary approach to treat congenital malformations of neural tubes.

The Modified Bushen Antai Recipe Upregulates Estrogen and Progesterone Receptors at the Maternal-Fetal Interface in Pregnant Rats with Mifepristone-Induced Pregnancy Loss.

BACKGROUND: The modified Bushen Antai recipe (BSAT) is a centuries-old traditional Chinese medicine that we use in our center as a therapy against pregnancy loss. Our study aimed to explore the potential benefit and mechanism of BSAT in pregnant rats with mifepristone-induced pregnancy loss. MATERIALS AND METHODS: The signature compounds of the eight BSAT ingredients were analyzed by high-performance liquid chromatography (HPLC). The BSAT group (n = 8) was treated daily with 6.3 ml/kg BSAT from gestation day (D) 0.5 to 10.5 and once with 1.25 mg/kg mifepristone on D 10.5. Normal saline replaced BSAT in the model group (n = 8), and both BSAT and mifepristone in the control group (n = 8). Morphological and histological analyses were performed on D 13.5. RESULTS: BSAT contains eight medicinal ingredients including Cuscuta chinensis and Dipsacus asperoides. The HPLC analysis detected the signature compounds of seven medicinal ingredients in the extract. Embryo resorption rate in the BSAT group was significantly lower than that in the model group, although the number of surviving embryos was similar between the two groups. Hematoxylin and eosin (HE) staining suggested that the maximum cross-sectional area of the placenta and the area ratio of the placental labyrinth in the BSAT group were higher than those in the model group. Immunohistochemical (IHC) staining indicated that the expression of ki67, estrogen receptor alpha (ERα), and progesterone receptor (PR) in the placental labyrinth of the BSAT group was higher than that of the model group. Furthermore, the protein levels of ERα, PR, phospho-Akt/Akt, and phospho-Erk1/2/Erk1/2 in the BSAT group were higher than those in the control group. The mRNA levels of ERα and PR in the BSAT group were higher than those in the control group. CONCLUSIONS: BSAT may induce estrogen and progesterone receptors by phosphorylation via the classic Akt and Erk1/2 signaling pathways in the maternal-fetal interface of pregnant rats, thereby reducing the pregnancy loss rate and improving the live birth rate.

Formate rescues neural tube defects caused by mutations in Slc25a32.

Periconceptional folic acid (FA) supplementation significantly reduces the prevalence of neural tube defects (NTDs). Unfortunately, some NTDs are FA resistant, and as such, NTDs remain a global public health concern. Previous studies have identified SLC25A32 as a mitochondrial folate transporter (MFT), which is capable of transferring tetrahydrofolate (THF) from cellular cytoplasm to the mitochondria in vitro. Herein, we show that gene trap inactivation of Slc25a32 (Mft) in mice induces NTDs that are folate (5-methyltetrahydrofolate, 5-mTHF) resistant yet are preventable by formate supplementation. Slc25a32gt/gt embryos die in utero with 100% penetrant cranial NTDs. 5-mTHF supplementation failed to promote normal neural tube closure (NTC) in mutant embryos, while formate supplementation enabled the majority (78%) of knockout embryos to complete NTC. A parallel genetic study in human subjects with NTDs identified biallelic loss of function SLC25A32 variants in a cranial NTD case. These data demonstrate that the loss of functional Slc25a32 results in cranial NTDs in mice and has also been observed in a human NTD patient.

[Identification characters of leaf morphological and venation pattern of Abutilon indicum with its confused herb A. theophrasti].

OBJECTIVE: To study the identification characters of Abutilon indicum and its confused herb A. theophrasti and establish an identification method. METHOD: LMVP (leaf morphological-venation pattern for identification Chinese herbs), and QAERM (quantitatively analyze and evaluate reliability for the method of identification Chinese herbs) were applied for the study. RESULT: The main differences of the leaf of Abutilon indicum: leaf margin cilia short and not clear (75-200 microm), petiole top with long straight hairs (up to 1.0-1.2 cm), when the tertiary veins and intersecondary veins issue from the lower part of midrib, their angles of divergence usually were wide acute (65 degrees-80 degrees) and a few was about right angle (80 degrees-100 degrees). The main differences of the leaf of A. theophrasti: leaf margin cilia long and clear (200-500 microm), petiole top with short straight hairs (most 0.1-0.5 mm), when the tertiary veins and intersecondary veins issue from the lower part of midrib, their angles of divergence usually were about right angle (80 degrees-100 degrees) and a few was wide acute (65 degrees-80 degrees). With the mentioned three groups of key differences, the both plants could be successfully identified from each other. The accuracy of identification results (AC) was from 97.5% to 99.6%, the repeatability of identification results: agreement rate for observation (ARO) was 97.1% and Kappa value was 0.94. CONCLUSION: The established method is simple, rapid, economic and reliable.

[Query: distinguish between dicot and monocot according to whether free vein termination exist or not].

OBJECTIVE: To observe and study the correctness of the proposition dicot have free vein termination but monocot have not. To evaluate the reliability of the identification principle that is based on above proposition. METHOD: Fourteen species of pharmaceutical monocot come from 9 families and 3 species of pharmaceutical dicot come from 3 families were observed by using the method of microscopic identification. RESULT: The free vein termination existed in the vein-islets of 10 species of monocot from 8 families, Zingiberaceae, Stemonaceae, Gramineae, Liliaceae, Orchidaceae, Alismataceae, Dioscoreaceae and Araceae ect, and it was not to find the free vein termination in monocot Iris tectorum, Belamcanda chinensis (Fam. Iridaceae) and Imperata cylindrica, Coix lacryma-jobi (Fam. Gramineae). Three species dicot, Cinnamomum burmannii, Castanea mollissima, Mangifera indica, all had the free vein termination. The free vein termination of C. burnmannii was sparse, and C. mollissima's was usually covered by non-glandular hair. CONCLUSION: The proposition is not universally applicable, monocot have no free vein termination. The identification principle is unreliable, which distinguish between medicinal dicot and monocot leaves according to whether the free vein termination exists or not.

[Long-term outcomes of individualized biofeedback training based on the underlying dysfunction for patients with imperforate anus].

OBJECTIVE: Although better anorectal function was achieved due to the advanced operation procedures, the fecal incontinence and constipation still remain as major postoperative complications. The aim of this study was to design a individualized biofeedback program based on the underlying dysfunctions and to assess the short-term and long-term clinical and quality of life outcomes for patients with imperforate anus. METHODS: Thirty-one patients were treated using individualized biofeedback program according to their different dysfunctions after completed assessment of anorectal function. The many of special biofeedback training methods that consisted of the strengthening perianal muscles biofeedback, sensory discrimination biofeedback, synchronizing external sphincter contraction biofeedback and improving defecation dynamic biofeedback were selected to make up the individualized biofeedback training program. The clinical scores and quality of life scores were assessed at short-term and long-term follow-up. RESULTS: The clinical scores and quality of life scores in patients at before training group, after biofeedback training group, short-term follow-up group and long-term follow-up group were 3.2 +/- 1.1 and 8.7 +/- 1.1, 5.2 +/- 0.6 and 11.0 +/- 0.8, 5.0 +/- 0.6 and 10.8 +/- 0.9, and 4.6 +/- 0.7 and 10.0 +/- 0.7 respectively. After biofeedback training, the first leak volume and all parameters that evaluated the function of the perianal muscles were increased significantly, the abnormal rectal threshold sensations and defecation dynamics were reverted to normal, and 82% patients who have not external anal sphincter reflex acquired the new reflex. The latencies of pudendo-anal reflex have significant difference in patients with poor response compared with those patients with good response before and after the biofeedback training. At short-term follow-up 12 patients (57%) maintained the clinical outcome and only 9 patients (43%) have regressions slightly. At long-term follow-up 7 patients (33%) maintained the clinical outcome very well and 14 patients (67%) have regressions. Although the clinical and quality of life scores have decreased slightly at follow-up, they were still increased significantly compared with before biofeedback training group. CONCLUSION: The causes of postoperative fecal dysfunction in patients with imperforate anus are multifactorial, and the individualized biofeedback is more suitable for improvement of the clinical outcome and the quality of life, and it maintain good clinical outcome and quality of life at short-term and long-term follow-up.