Construction of a cross-species cell landscape at single-cell level.

Individual cells are basic units of life. Despite extensive efforts to characterize the cellular heterogeneity of different organisms, cross-species comparisons of landscape dynamics have not been achieved. Here, we applied single-cell RNA sequencing (scRNA-seq) to map organism-level cell landscapes at multiple life stages for mice, zebrafish and Drosophila. By integrating the comprehensive dataset of > 2.6 million single cells, we constructed a cross-species cell landscape and identified signatures and common pathways that changed throughout the life span. We identified structural inflammation and mitochondrial dysfunction as the most common hallmarks of organism aging, and found that pharmacological activation of mitochondrial metabolism alleviated aging phenotypes in mice. The cross-species cell landscape with other published datasets were stored in an integrated online portal-Cell Landscape. Our work provides a valuable resource for studying lineage development, maturation and aging.

How many cell types are there in nature? How do they change during the life cycle? These are two fundamental questions that researchers have been trying to understand in the area of biology. In this study, single-cell mRNA sequencing data were used to profile over 2.6 million individual cells from mice, zebrafish and Drosophila at different life stages, 1.3 million of which were newly collected. The comprehensive datasets allow investigators to construct a cross-species cell landscape that helps to reveal the conservation and diversity of cell taxonomies at genetic and regulatory levels. The resources in this study are assembled into a publicly available website at http://bis.zju.edu.cn/cellatlas/.

Genetically engineering of Saccharomyces cerevisiae for enhanced oral delivery vaccine vehicle.

As a series of our previous studies reported, recombinant yeast can be the oral vaccines to deliver designed protein and DNA, as well as functional shRNA, into dendritic cells (DCs) in mice for specific immune regulation. Here, we report the further optimization of oral yeast-based vaccine from two aspects (yeast characteristics and recombinant DNA constitution) to improve the effect of immune regulation. After screening four genes in negative regulation of glucan synthesis in yeast (MNN9, GUP1, PBS2 and EXG1), this research combined HDR-based genome editing technology with Cre-loxP technology to acquire 15 gene-knockout strains without drug resistance-gene to exclude biosafety risks; afterward, oral feeding experiments were performed on the mice using 15 oral recombinant yeast-based vaccines constructed by the gene-knockout strains harboring pCMV-MSTN plasmid to screen the target strain with more effective inducing mstn-specific antibody which in turn increasing weight gain effect. And subsequently based on the selected gene-knockout strain, the recombinant DNA in the oral recombinant yeast-based vaccine is optimized via a combination of protein fusion expression (OVA-MSTN) and interfering RNA technology (shRNA-IL21), comparison in terms of both weight gain effect and antibody titer revealed that the selected gene-knockout strain (GUP1ΔEXG1Δ) combined with specific recombinant DNA (pCMV-OVA-MSTN-shIL2) had a better effect of the vaccine. This study provides a useful reference to the subsequent construction of a more efficient oral recombinant yeast-based vaccine in the food and pharmaceutical industry.

Exploring the emergence of morphological asymmetries around the brain's Sylvian fissure: a longitudinal study of shape variability in preterm infants.

Brain folding patterns vary within the human species, but some folding properties are common across individuals, including the Sylvian fissure's inter-hemispheric asymmetry. Contrarily to the other brain folds (sulci), the Sylvian fissure develops through the process of opercularization, with the frontal, parietal, and temporal lobes growing over the insular lobe. Its asymmetry may be related to the leftward functional lateralization for language processing, but the time course of these asymmetries' development is still poorly understood. In this study, we investigated refined shape features of the Sylvian fissure and their longitudinal development in 71 infants born extremely preterm (mean gestational age at birth: 26.5 weeks) and imaged once before and once at term-equivalent age (TEA). We additionally assessed asymmetrical sulcal patterns at TEA in the perisylvian and inferior frontal regions, neighbor to the Sylvian fissure. While reproducing renowned strong asymmetries in the Sylvian fissure, we captured an early encoding of its main asymmetrical shape features, and we observed global asymmetrical shape features representative of a more pronounced opercularization in the left hemisphere, contrasting with the previously reported right hemisphere advance in sulcation around birth. This added novel insights about the processes governing early-life brain folding mechanisms, potentially linked to the development of language-related capacities.

Clinical effects of cocktail injection on the thoracolumbar fascia injury during percutaneous vertebroplasty for osteoporotic vertebral compression fractures: a single-center, retrospective case-control study.

BACKGROUND: Nowadays, there is a lack of effective intraoperative treatment for thoracolumbar fascia injury (TFI) of osteoporotic vertebral compression fractures (OVCFs), which may lead to postoperative residual pain. We aimed to evaluate the clinical effects of cocktail injection on the TFI during percutaneous vertebroplasty (PVP) for OVCFs. METHODS: A retrospective study of OVCFs with TFI underwent PVP with cocktail injection (Cocktail group, 58 cases) or PVP (Routine group, 64 cases) was conducted. The surgical outcomes, visual analog scale (VAS) score, oswestry disability index (ODI), incidence of residual pain at 1 day and 7 days postoperatively, the rate and duration of taking painkillers during 7 days postoperatively after PVP were compared between them. RESULTS: No differences in baseline data, volume of bone cement injected and bone cement leakage were observed between the two groups, while the operation time of the routine group (44.3 ± 7.8 min) was less than that (47.5 ± 9.1 min) of the cocktail group (P < 0.05). However, the VAS scores (2.4 ± 0.8, 2.2 ± 0.7), ODI (25.2 ± 4.2, 22.3 ± 2.9), the incidence of residual pain (8.6%, 3.4%) at 1 and 7 days postoperatively, the rate (6.9%) and duration ( 2.5 ± 0.6 ) of taking painkillers during 7 days postoperatively in the cocktail group were better than those (3.4 ± 1.0, 2.9 ± 0.7, 34.1 ± 4.7, 28.6 ± 3.6, 23.4%, 15.6%, 28.1%, 4.2 ± 1.4) in the routine group (P < 0.05), respectively. CONCLUSION: PVP combined with cocktail injection increased the operation time in the treatment of OVCFs with TFI, but it can more effectively relieve pain, reduce the risk of residual pain at 1 day and 7 days postoperatively, and decrease the use and duration of taking painkillers.

TIMP2 mediates endoplasmic reticulum stress contributing to sepsis-induced acute kidney injury.

Tissue inhibitor of metalloproteinase 2 (TIMP2) has been recognized as an important biomarker for predicting acute kidney injury (AKI) because of its involvement in the process of inflammation and apoptosis in septic AKI. Endoplasmic reticulum (ER) stress, a condition of disrupted ER homeostasis, is implicated in multiple pathophysiological processes, including kidney disease. Herein, we investigated the correlation between ER stress and septic AKI and further explored how TIMP2 regulated ER stress-mediated apoptosis. To assess the role of TIMP2 in sepsis-induced AKI, we used a cecal ligation and puncture (CLP) model in mice with tubule-specific deficiency of TIMP2 (Ksp-Cre/TIMP2flox/flox ) and their wild-type counterparts. Compared to the wild-type mice, TIMP2-deficient mice demonstrated lower serum creatinine levels and decreased ER stress-mediated apoptosis when subjected to CLP. Interestingly, in human kidney (HK-2) cells, overexpression of TIMP2 caused ER stress, whereas TIMP2 knockdown attenuated lipopolysaccharide-induced ER stress and apoptosis. TIMP2 interacted with the binding immunoglobulin protein, an ER chaperone, and facilitates its extracellular secretion, thereby triggering ER stress. This study identified that the deletion of TIMP2 in mouse tubules mitigated sepsis-induced AKI by inhibiting ER stress-mediated apoptosis, which might be a potential therapeutic strategy to alleviate renal injury.

Comparison of clinical effects of percutaneous vertebroplasty with two different puncture approaches on the treatment of thoracolumbar osteoporotic vertebral compression fractures with narrow pedicles: a retrospective controlled study.

PURPOSE: To evaluate the effects of percutaneous vertebroplasty (PVP) with conventional transpedicle approach (CTA) or basal transverse process-pedicle approach (BTPA) on the treatment of thoracolumbar osteoporotic vertebral compression fractures (TL-OVCFs) with narrow pedicles. METHODS: A retrospective study of TL-OVCFs with narrow pedicles was performed, including 78 cases of CTA and 84 cases of BTPA. The surgical outcomes, radiographic parameters [the width and height of the pedicle (PW, PH), the inclination angle of puncture (PIA)] and clinical indicators [visual analog scale (VAS) score, Oswestry Disability Index (ODI)] of two groups were compared. RESULTS: In terms of surgical outcomes of them, there was no difference in operation time (P > 0.05), while the volume of bone cement, the incidence of bone cement leakage and rate of good bone cement distribution were significantly worse in the CTA group (4.4 ± 0.6 ml vs. 5.5 ± 0.5 ml, 37.2% vs. 20.2%, 52.6% vs. 79.8%, P < 0.05). As for radiographic parameters and clinical indicators of them, the differences were not observed in the PH, PW, preoperative VAS score and ODI (P > 0.05), whereas the PIA, VAS score and ODI at 1 day postoperatively were significantly better in the BTPA group (17.3 ± 2.1° vs. 29.6 ± 2.8°, 2.7 ± 0.7 vs. 2.1 ± 0.8, 32.8 ± 4.6 vs. 26.7 ± 4.0, P < 0.05). CONCLUSION: The study provided solid evidence that PVP with BTPA had more advantages in the treatment of TL-OVCFs with narrow pedicles, which can better relieve postoperative pain.

Disentangling the variability of the superficial white matter organization using regional-tractogram-based population stratification.

Each variation of the cortical folding pattern implies a particular rearrangement of the geometry of the fibers of the underlying white matter. While this rearrangement only impacts the ends of the long pathways, it may affect most of the trajectory of the short bundles. Therefore, mapping the short fibers of the human brain using diffusion-based tractography requires a dedicated strategy to overcome the variability of the folding patterns. In this paper, we propose a fiber-based stratification strategy splitting the population into homogeneous groups for disentangling the superficial white matter bundle organization. This strategy introduces a new refined fiber distance which includes angular considerations for inferring fine-grained atlases of the short bundles surrounding a specific sulcus and a subtractogram distance that quantifies the similitude between fiber sets of two different subjects. The stratification splits the population into groups with similar regional fiber organization using manifold learning. We first successfully test the hypothesis that the main source of variability of the regional fiber organization is the variability of the regional folding pattern. Then, in each group, we proceed with the automatic identification of the most stable bundles, at a higher granularity level than what can be achieved with the non-stratified whole population, enabling the disentanglement of the very variable configuration of the short fibers. Finally, the method searches for bundle correspondence across groups to build a population level atlas. As a proof of concept, the atlas refinement achieved by this strategy is illustrated for the fibers that surround the central sulcus and the superior temporal sulcus using the HCP dataset.

Shape variability of the central sulcus in the developing brain: A longitudinal descriptive and predictive study in preterm infants.

Despite growing evidence of links between sulcation and function in the adult brain, the folding dynamics, occurring mostly before normal-term-birth, is vastly unknown. Looking into the development of cortical sulci in infants can give us keys to address fundamental questions: what is the sulcal shape variability in the developing brain? When are the shape features encoded? How are these morphological parameters related to further functional development? In this study, we aimed to investigate the shape variability of the developing central sulcus, which is the frontier between the primary somatosensory and motor cortices. We studied a cohort of 71 extremely preterm infants scanned twice using MRI - once around 30 weeks post-menstrual age (w PMA) and once at term-equivalent age, around 40w PMA -, in order to quantify the sulcus's shape variability using manifold learning, regardless of age-group or hemisphere. We then used these shape descriptors to evaluate the sulcus's variability at both ages and to assess hemispheric and age-group specificities. This led us to propose a description of ten shape features capturing the variability in the central sulcus of preterm infants. Our results suggested that most of these features (8/10) are encoded as early as 30w PMA. We unprecedentedly observed hemispheric asymmetries at both ages, and the one captured at term-equivalent age seems to correspond with the asymmetry pattern previously reported in adults. We further trained classifiers in order to explore the predictive value of these shape features on manual performance at 5 years of age (handedness and fine motor outcome). The central sulcus's shape alone showed a limited but relevant predictive capacity in both cases. The study of sulcal shape features during early neurodevelopment may participate to a better comprehension of the complex links between morphological and functional organization of the developing brain.

Gut microbiota-testis axis: FMT improves systemic and testicular micro-environment to increase semen quality in type 1 diabetes.

BACKGROUND: Clinical data suggest that male reproductive dysfunction especially infertility is a critical issue for type 1 diabetic patient (T1D) because most of them are at the reproductive age. Gut dysbiosis is involved in T1D related male infertility. However, the improved gut microbiota can be used to boost spermatogenesis and male fertility in T1D remains incompletely understood. METHODS: T1D was established in ICR (CD1) mice with streptozotocin. Alginate oligosaccharide (AOS) improved gut microbiota (fecal microbiota transplantation (FMT) from AOS improved gut microbiota; A10-FMT) was transplanted into the T1D mice by oral administration. Semen quality, gut microbiota, blood metabolism, liver, and spleen tissues were determined to investigate the beneficial effects of A10-FMT on spermatogenesis and underlying mechanisms. RESULTS: We found that A10-FMT significantly decreased blood glucose and glycogen, and increased semen quality in streptozotocin-induced T1D subjects. A10-FMT improved T1D-disturbed gut microbiota, especially the increase in small intestinal lactobacillus, and blood and testicular metabolome to produce n-3 polyunsaturated fatty acid (PUFA) docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) to ameliorate spermatogenesis and semen quality. Moreover, A10-FMT can improve spleen and liver functions to strengthen the systemic environment for sperm development. FMT from gut microbiota of control animals (Con-FMT) produced some beneficial effects; however, to a smaller extent. CONCLUSIONS: AOS-improved gut microbiota (specific microbes) may serve as a novel, promising therapeutic approach for the improvement of semen quality and male fertility in T1D patients via gut microbiota-testis axis.

The mutation features and geographical distributions of the surface glycoprotein (S gene) in SARS-CoV-2 strains: A comparative analysis of the early and current strains.

The surface glycoprotein (S protein) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was used to develop coronavirus disease 2019 (COVID-19) vaccines. However, SARS-CoV-2, especially the S protein, has undergone rapid evolution and mutation, which has remained to be determined. Here, we analyzed and compared the early (12 237) and the current (more than 10 million) SARS-CoV-2 strains to identify the mutation features and geographical distribution of the S gene and S protein. Results showed that in the early strains, most of the loci were with relative low mutation frequency except S: 23403 (4486 strains), while in the current strains, there was a surge in the mutation strains and frequency, with S: 23403 constantly being the highest one, but tremendously increased to approximately 1050 times. Furthermore, D614 (S: 23403) was one of the most highly frequent mutations in the S protein of Omicron as of March 2022, and most of the mutant strains were still from the United States, and the United Kingdom. Further analysis demonstrated that in the receptor-binding domain, most of the loci with low mutation frequency in the early strains, while S: 22995 was nowadays the most prevalent loci with 3 122 491 strains in the current strains. Overall, we compare the mutation features of the S region in SARS-CoV-2 strains between the early and the current stains, providing insight into further studies in concert with emerging SARS-CoV-2 variants for COVID-19 vaccines.

14-3-3ζ binds to and stabilizes phospho-beclin 1S295 and induces autophagy in hepatocellular carcinoma cells.

Data from The Cancer Genome Atlas (TCGA) indicate that the expression levels of 14-3-3ζ and beclin 1 (a key molecule involved in cellular autophagy) are up-regulated and positively correlated with each other (R = .5, P < .05) in HCC tissues. Chemoresistance developed in hepatoma cancer cells is associated with autophagy initiation. This study aimed to explore 14-3-3ζ's role in regulating autophagy in HCC cells, with a focus on beclin 1. The co-localization of 14-3-3ζ and beclin 1 was detectable in primary HCC tissues. To simulate in vivo tumour microenvironment (hypoxia), CSQT-2 and HCC-LM3 cells were exposed to 2% oxygen for 24 hours. The protein levels of 14-3-3ζ and phospho-beclin 1S295 peaked at 12 hours following hypoxia. Meanwhile, the strongest autophagy flux occurred: LC3II was increased, and p62 was decreased significantly. By sequencing the coding area of BECN 1 gene of CSQT-2 and HCC-LM3 cells, we found that the predicted translational products of BECN 1 gene contained RLPS295 VP (R, arginine; L, leucine; P, proline; S, serine; V, valine), a classic 14-3-3ζ binding motif. CO-IP results confirmed that 14-3-3ζ bound to beclin 1, and this connection was markedly weakened when S295 was mutated into A295 (alanine). Further, 14-3-3ζ overexpression prevented phospho-beclin 1S295 from degradation and enhanced its binding to VPS34, whilst its knockdown accelerated the degradation. Additionally, 14-3-3ζ enhanced the chemoresistance of HCC cells to cis-diammined dichloridoplatium by activating autophagy. Our work reveals that 14-3-3ζ binds to and stabilizes phospho-beclin 1S295 and induces autophagy in HCC cells to resist chemotherapy.

Early administration of amino acids with different doses in low birth weight premature infants.

BACKGROUND: The reasonable use of amino acids (AAs) in parenteral nutrition (PN) is very critical to the growth and development of premature infants. However, the appropriate dose of AAs has not been determined. Our study was designed to investigate the clinical effect of two different doses of AAs in PN for low birth weight premature infants. MATERIALS AND METHODS: This randomized controlled study included 191 preterm infants who admitted to the neonatal intensive care unit of the First Affiliated Hospital of Nanjing Medical University from June 2015 to December 2016 and they were randomly divided into Group 1 (n = 81) and Group 2 (n = 110). In Group 1, the starting dose of AAs dose was 1.0-1.5 g/kg/day, which was increased by 0.5 g/kg with the maximum dose at 3.5 g/kg/day. In Group 2, the starting dose of AAs was 1.8-2.5 g/kg/day and was increased by 1.0 g/kg with the maximum dose at 4.0-4.5 g/kg/day. We analyzed the clinical characteristics, body weight, body length, total calorie intake, nonprotein calorie intake, total protein intake, liver and kidney function, and complications of the two groups of preterm infants. RESULTS: The start of enteral feeding and the recovery of birth weight in Group 2 were earlier than those in Group 1 (3.83 ± 3.15 day vs. 5.53 ± 5.63 day, P = 0.016 and 6.36 ± 4.88 day vs. 8.48 ± 9.27 day, P = 0.043, respectively). The duration of PN and the time before total enteral nutrition were shorter in Group 2 than in Group 1 (16.46 ± 10.33 day vs. 21.41 ± 18.00 day, P = 0.029 and 15.47 ± 10.54 day vs. 19.47 ± 14.57 day, P = 0.038; respectively). The duration of mechanical ventilation (1.12 ± 2.62 day vs. 3.31 ± 8.13 day, P = 0.028) in Group 2 was shorter than that in Group 1. CONCLUSION: High doses of AAs in the early PN for preterm infants facilitate the promotion of early growth and development, advance recovery of birth weight, reduce the duration of PN, and reduce respiratory support without increasing the incidence of complications.

Effects of activin A on the transcriptome of mouse oogenesis in vitro.

From the previous research, it has been supported that activin A (ActA) is conducive to ovarian development in vitro. In the present paper, with the aim to identify the molecular pathways through which ActA can influence processes of the fetal and early postnatal oogenesis, we analyzed the transcriptome of embryonic ovaries (12.5 days postcoitum) in vitro cultured with or without ActA for 6 days, as well as the produced oocytes for 28 days, and further compared the gene expression profile with their in vivo counterparts. With the confirmation of designed test, we found that the addition of ActA to the ovary culture tended, generally, to align oocyte gene expression to the in vivo condition, in particular of a number of genes involved in meiosis and epigenetic modifications of histones. In particular, we identified DNA recombination during the oocyte meiotic prophase I and lysine trimethylation of the histone H3K27 during the oocyte growth phase as molecular pathways modulated by ActA.

Role of high-mobility group B1 in myocardial injury induced by coronary microembolization in rats.

OBJECTIVE: This study aimed to explore the effects of high-mobility group B1 (HMGB1) on coronary microembolization (CME)-induced myocardial inflammation, myocardial apoptosis, and cardiac function injury in rats. METHODS: Forty Sprague-Dawley rats were divided into sham operation group (sham group), microembolization group (CME group), CME + HMGB1 siRNA (HMGB1 siRNA) group, and CME + scrambled siRNA (control siRNA) group (10 rats in each group). The CME model group was constructed by injecting microembolism spheres into the apex of the left ventricle after clamping the ascending aorta. The sham group was constructed by injecting the same amount of saline. The HMGB1 siRNA group was injected with HMGB1 siRNA transfection complex via the tail vein 72 hours before CME modeling. The control siRNA group was injected with the same amount of scrambled siRNA mixture through the tail vein 72 hours before CME modeling. The cardiac function, serum cardiac troponin I level, and apoptotic index were examined 12 hours after the surgery. The levels of HMGB1, nuclear factor-κB (NF-κB) p65, glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), cleaved caspase-12, cleaved caspase-3, tumor necrosis factor-α (TNF-α), and interleukin 1ß (IL-1ß) were detected. RESULTS: Myocardial dysfunction, enhanced serum cardiac troponin I level, and apoptotic index were induced following CME. Moreover, CME increased the expression of HMGB1, NF-κB p65, GRP78, CHOP, cleaved caspase-12, cleaved caspase-3, TNF-α, and IL-1ß. HMGB1 siRNA reversed these effects, whereas scrambled siRNA had no effect. CONCLUSIONS: Inhibition of HMGB1 expression reduced CME-induced myocardial injury and improved cardiac function. Hence, it may serve as a new target for preventing and treating the CME-induced myocardial injury.

Identification of oxidative stress-related Xdh gene as a di(2-ethylhexyl)phthalate (DEHP) target and the use of melatonin to alleviate the DEHP-induced impairments in newborn mouse ovaries.

This study, using an in vitro ovary culture model, investigates the mechanisms through which di(2-ethylhexyl)phthalate (DEHP) impairs germ cell cyst breakdown and primordial follicle assembly. The results indicate the latter effects exerted by 10 or 100 µmol/L DEHP in cultured newborn ovaries were associated with increased levels of reactive oxygen species (ROS) and apoptosis. Based on a transcriptome analysis, we found the expression of the oxidative stress-related gene Xdh (xanthine dehydrogenase) was significantly upregulated in DEHP-cultured ovaries. Two treatments, namely Xdh RNAi or the addition of melatonin to the ovary culture, inhibited the increase in Xdh expression and ROS levels caused by DEHP and, at the same time, reduced apoptosis and the impairment of primordial follicle assembly in the treated ovaries. Together, the results identify Xdh gene as one of the major targets of DEHP in newborn ovaries and that the consequent increased level of ROS is possibly responsible for the increment of apoptosis and primordial follicle assembly impairment. At the same time, they highlight that melatonin alleviates the effects of DEHP as with other endocrine-disrupting compounds on the ovary.

The role of miR-431-5p in regulating pulmonary surfactant expression in vitro.

BACKGROUND: Pulmonary surfactant is the complex mixture of lipid and protein that covers the alveolar surface. Pulmonary surfactant deficiency is one of the main causes of neonatal respiratory distress. Recent studies showed that miRNA plays an important role in lung development, but research into miR-431 regulation of pulmonary surfactant are sparse. In this study, we explored the regulatory role of miR-431-5p in the expression of pulmonary surfactant and identified its potential target gene, Smad4. METHODS: The bioinformatics tool TargetScan was used to predict the targets of miR-431. The expression of miR-431-5p was achieved via transfection of miR-431-5p mimics, an miR-431-5p inhibitor and corresponding negative control. The level of miR-431-5p was determined using quantitative real-time PCR. The CCK8 assay was conducted to confirm cell growth 12 h after transfection with miR-431-5p mimics, inhibitor or NC. Smad4 and surfactant-associated proteins in A549 were analyzed using western blot and quantitative real-time PCR. RESULTS: Smad4 was validated as a target of miR-431 in A549 cells. Overexpression of miR-431 accelerated A549 proliferation and inhibited A549 apoptosis. The mRNA and protein levels for the surfactant proteins (SP-A, SP-B, SP-C and SP-D) were found to be differentially expressed in A549 cells over- or under-expressing miR-431-5p. CONCLUSION: Our results show that miR-431-5p is critical for pulmonary surfactant expression and that its regulation is closely related to the TGF-ß/Smad4 pathway. These results will help us to study the pathophysiological mechanism of lung developmental diseases.

[Sun's coronal sulcus ring incision plus suprapubic liposuction versus suprapubic V-Y incision for penile elongation].

OBJECTIVE: To investigate the clinical effect of suprapubic V-Y incision versus that of Sun's coronal sulcus ring incision plus suprapubic liposuction (CSRI+SPLS) for penile elongation. METHODS: From December 2010 to January 2018, 100 adult males with congenital short penis underwent suprapubic V-Y incision (the V-Y group, n = 50) or CSRI+SPLS (n = 50) for penile elongation surgery in our department. We statistically analyzed the clinical data on the two groups of patients, including age, body mass index (BMI), pre- and post-operative penile lengths in flaccid and erectile states, operation time, intraoperative blood loss, postoperative use of analgesics, postoperative hospital stay, stage-Ⅰ wound healing, incidence of postoperative complications, and quality of pre- and post-operative sexual life. RESULTS: The average ages of the patients in the V-Y and CSRI+SPLS groups were 23.5 and 23.0 years, their BMIs (26.59 ± 1.16) and (26.44 ± 0.96) kg/m2, preoperative flaccid penile lengths (5.11 ± 0.30) and (5.12 ± 0.35) cm, preoperative erectile penile lengths (7.57 ± 0.65) and (7.35 ± 0.59) cm, postoperative flaccid penile lengths (7.80 ± 0.40) and (7.79 ± 0.42) cm, postoperative erectile penile lengths (11.59 ± 0.55) and (11.47 ± 0.64) cm, none with statistically significant difference between the two groups (P > 0.05). Compared with the V-Y group, the CSRI+SPLS group showed a markedly shorter operation time (108 ï¼»90－120ï¼½ vs 51 ï¼»45－58ï¼½ min, P < 0.01), less intraoperative blood loss (30 ï¼»15－45ï¼½ vs 15 ï¼»5－25ï¼½ ml, P < 0.01), shorter postoperative hospital stay 8 ï¼»6－11ï¼½ vs 4 ï¼»2－6ï¼½ d, P < 0.01), lower incidence rates of postoperative hematoma (23.33% ï¼»7 casesï¼½ vs 0, P < 0.05) and infection (20.00% ï¼»6 casesï¼½ vs 0, P < 0.05), higher stage-Ⅰ healing rate (76.67% ï¼»23 casesï¼½ vs 100.00% ï¼»30 casesï¼½, P < 0.05), lower rate of postoperative use of analgesics (26.67% ï¼»8 casesï¼½ vs 3.33% ï¼»1 caseï¼½, P < 0.05), and higher score on the quality of postoperative sexual life (36.73 ± 5.41 vs 42.07 ± 3.64, P < 0.01) though with no statistically significant difference preoperatively (28.70 ± 4.87 vs 28.27 ± 3.40, P > 0.05). CONCLUSIONS: As a surgical procedure for penile elongation, Sun's coronal sulcus ring incision plus suprapubic liposuction is superior to suprapubic V-Y incision for its advantages of lower invasiveness, shorter operation time, less intraoperative blood loss, and lower incidence of postoperative complications.

[Urolastic: A new medical liquid silicone rubber elastomer compound for penile augmentation in beagle dogs].

OBJECTIVE: To study the safety, durability and effectiveness of a new medical liquid silicone rubber elastomer (LSRE) compound Urolastic in penile augmentation in beagle dogs. METHODS: A total of 30 beagle dogs were included in the experiment. The diameters of the proximal, middle and distal penis of the animals were measured and the indexes of blood routine, liver and kidney function and electrolytes obtained before and at 1 week and 3 months after penile subcutaneous injection of the LSRE compound. CT scanning and pathological examinations of the liver, kidney and penile tissues were performed at 1 and 3 months after treatment. RESULTS: The diameters of the proximal, middle and distal penis of the dogs were increased by (0.4 ± 0.3) cm, (0.6 ± 0.1) cm and (0.5 ± 0.3) cm at 1 week, and (0.4 ± 0.2) cm, (0.5 ± 0.1) cm and (0.6 ± 0.2) cm at 3 months after injection of the LSRE compound, with statistically significant difference from the baseline (P < 0.01 or P < 0.05) but not between the 1st week and the 3rd month (P > 0.05). The counts of leukocytes and neutrophils were markedly increased compared with the baseline (ï¼»18.16 ± 2.57ï¼½ vs ï¼»15.16 ± 3.17ï¼½ g/L, P < 0.05; ï¼»77.34 ± 9.21ï¼½% vs ï¼»67.18 ± 8.25ï¼½%, P < 0.05), but not the other blood routine indexes. There were no statistically significant differences in the liver and kidney functions or electrolytes before and after the injection. At 1 and 3 months after treatment, Urolastic was clearly visible and the injection points were irregular in shape at CT imaging. The anatomical findings were consistent with the CT manifestations and showed that the material was easily separated from the surrounding tissues. No significant inflammatory cell infiltration was observed in pathological examinations at 1 and 3 months. CONCLUSIONS: The new medical LSRE compound Urolastic has a good clinical application prospect in penile augmentation for its advantages of significant effectiveness and high safety.

[Continuous expression of miR-431 during lung development in Sprague-Dawley rats].

OBJECTIVE: To study the role of miR-431 in lung development and morphology. METHODS: According to the stage of lung development in rats, Sprague-Dawley rats at embryonic day 16 (E16), embryonic day (E19), embryonic day (E21), postnatal day 1 (P1), postnatal day 3 (P3), postnatal day 7 (P7), postnatal day 14 (P14) and 10 weeks after birth (P10 weeks) were selected, and lung tissue samples were collected for observation. Hematoxylin-eosin staining and transmission electron microscopy were performed to observe the morphology of lung tissue. Fluorescence in situ hybridization and real-time PCR were used to measure the expression of miR-431 during the critical stages of lung development (E19, E21 and P3). RESULTS: The E19 group had the formation of the lamellar body and type II alveolar epithelial cells in the fetal lung tissue. The number of lamellar bodies increased with the increasing gestational age, with aggregation and excretion. Pulmonary alveoli formed rapidly, the lung interstitium became thinner, and the microvascular system became mature after birth. Fluorescence in situ hybridization and real-time PCR showed that the expression of miR-431 gradually decreased with the increasing gestational age (P<0.05). CONCLUSIONS: The systematic and continuous morphological data of lung development is obtained in this experiment. In addition, miR-431 may play an important role in the negative regulation of lung development, which provides basis and direction for further research on the mechanism of lung development and related diseases.

Overexpression of PCK1 Gene Antagonizes Hepatocellular Carcinoma Through the Activation of Gluconeogenesis and Suppression of Glycolysis Pathways.

BACKGROUND/AIMS: Gluconeogenesis, a reverse process of glycolysis, is suppressed in neoplastic livers. Cytoplasmic phosphoenolpyruvate carboxykinase (PEPCK-C/PCK1, encoded by PCK1) is a step limiting enzyme of gluconeogenesis. The induced expression of the factor is reported to initiate gluconeogenesis process and antagonize hepatocellular carcinoma (HCC). In the current study, the effect of the modulation of PCK1 expression on HCC was assessed. METHODS: The levels of PCK1 in clinical HCC tissues and different HCC cell lines were investigated with real time quantitative PCR, immunochemistry, and western blotting. Thereafter, the expression of PCK1 gene was induced in two HCC cell lines and the effect of the overexpression on proliferation and migration potentials of HCC cells was detected with CCK-8 assay, flow cytometry, TUNEL staining, and transwell assay. The activities of glycolysis and gluconeogenesis pathways in PCK1-overexpressed HCC cell lines were detected with specific kits to underlie the mechanism by which PCK1 exerted its function. The results of the in vitro experiments were validated with HCC xenograft rat models. RESULTS: The expression levels of PCK1 were suppressed in HCC samples and in cells derived from HCC tissues. According to the results of the in vitro assays, the overexpression of PCK1 decreased viability, induced apoptosis, and inhibited migration in both HCC cell lines. The effect was associated with the suppressed glycolysis and the induced gluconeogenesis pathways, represented by the enhanced production of glucose and the limited production of pyruvic acid, lactate, citrate, and malate. The results of the in vitro assays were confirmed in rat models in that the growth rate of solid HCC tumors was reduced in mice transplanted with PCK1-overexpressed HCC cells. CONCLUSION: Findings outlined in the current study demonstrated that activating gluconeogenesis process via PCK1 overexpression was a potential treating strategy against HCC.