SHMT2 reduces fatty liver but is necessary for liver inflammation and fibrosis in mice.

Non-alcoholic fatty liver disease is associated with an irregular serine metabolism. Serine hydroxymethyltransferase 2 (SHMT2) is a liver enzyme that breaks down serine into glycine and one-carbon (1C) units critical for liver methylation reactions and overall health. However, the contribution of SHMT2 to hepatic 1C homeostasis and biological functions has yet to be defined in genetically modified animal models. We created a mouse strain with targeted SHMT2 knockout in hepatocytes to investigate this. The absence of SHMT2 increased serine and glycine levels in circulation, decreased liver methylation potential, and increased susceptibility to fatty liver disease. Interestingly, SHMT2-deficient mice developed simultaneous fatty liver, but when fed a diet high in fat, fructose, and cholesterol, they had significantly less inflammation and fibrosis. This study highlights the critical role of SHMT2 in maintaining hepatic 1C homeostasis and its stage-specific functions in the pathogenesis of NAFLD.

A Challenging Correlation between Tumor Cellularity and Somatic Variant Allele Fraction in Lung and Colorectal Cancers-Specimens of Low Tumor Percentage Should Be Analyzed with Caution.

Background and aims: The percentage of tumor cells (tumor cellularity) in a cancerous tissue has been assumed to correlate with the variant allele fraction (VAF) of an identified pathogenic variant. Many laboratories use the tumor cellularity as part of a quality criteria for specimen processing and clinical reporting. However, a systematic study of such correlation has yet to be shown. We performed a relatively large-scale study to determine whether pathologist-estimated tumor cellularity is correlated with next-generation sequencing (NGS)-derived VAF. Materials and Methods: A total of 1511 non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) specimens, including formalin-fixed paraffin-embedded (FFPE) and fine needle aspirated (FNA) tissues, were analyzed by cancer hotspot NGS. For a given specimen, pathogenic variants of BRAF, EGFR, KRAS, and NRAS were identified and the determined VAFs were correlated with the corresponding tissue tumor cellularity. Results: The coefficient of determination R-squared (R2) values were calculated for each correlation. All R2 values were lower than 0.25, indicating poor correlations. Pathogenic variants were found, not uncommonly, in tumor specimens that carried 10% or lower tumor cellularity. There were no apparent differences of R2 values between the FFPE and FNA specimens. Conclusion: In both NSCLC and CRC, the lack of linear relationship between tumor cellularity and VAF was found across a wide range of tumor cell percentages. Caution should be used when using tumor cellularity to triage specimens for NGS testing. The tumor cellularity should be considered in relation to the limit of detection of the specific assay for the proper interpretation of a negative test result.

Evaluation of pelvic structural abnormalities in primiparous women with stress urinary incontinence.

INTRODUCTION AND HYPOTHESIS: The objective was to evaluate the morphological characteristics of pelvic floor structure specific to de novo stress urinary incontinence (SUI) in primiparous women using three-dimensional (3D) reconstruction fusion technology based on static MRI combined with dynamic MRI. METHODS: Eighty-one primiparous women after the first vaginal delivery were studied, 40 with SUI and 41 without SUI. 3D reconstruction models based on static MRI were used to describe the anatomical abnormalities of pelvic floor tissues. Dynamic MRI was used to describe segmental activities of the urethra and vagina. The relationship between the morphometry and postpartum SUI was evaluated by logistic regression analysis and receiver operator characteristic curve. RESULTS: The differences in the distance from the bladder neck to the pubic symphysis (BSD), the angle between the posterior wall of the urethra and the anterior wall of the vagina, the width of the distal region of the vagina, urethral length, urethral compression muscle volume (CUV), and pubovisceral muscle volume, puborectal muscle volume, were measured, and except for the extremity of the anterior urethral wall, the total displacements (TDs) of the other sites between the two groups were statistically significant (p < 0.05). Logistic regression analysis showed that the BSD decreased, the CUV decreased, the TDs of the first site and the eighth site increment correlated significantly with postpartum SUI occurrence (p < 0.05). CONCLUSIONS: 3D reconstruction fusion technology provides an important support for a precise assessment of the pelvic floor dysfunction. The BSD, CUV, and iliococcygeus muscle volume have certain values in predicting de novo SUI after first vaginal birth.

Biomechanical study of the male lower urinary tract: Simulation of internal and external sphincters dyssynergia.

Urethral sphincter dysfunction is an important cause of stress urinary incontinence (SUI). The most effective treatment is the insertion of an artificial urethral sphincter (AUS), which relies to a large extent on the surgeon's experience. However, there is no quantitative standard for cuff tightness, resulting in frequent postoperative complications. This study aimed to investigate the effect of internal and external sphincter dyssynergia on urodynamic parameters in the lower urinary tract. A geometric model of male lower urinary tract tissue was constructed from collodion slices, accounting for the active behavior of the internal and external sphincters. Normal and dyssynergic internal and external sphincters (active sphincter behavior was individually injured by 25%, 50%, 75%, or 100%) were simulated with fluid-structure interactions and changes in urethral stress, displacement, and urine flow rate were detected. We found that when the internal sphincter was injured by 25%, 50%, 75%, and 100%, urethral stress near the internal sphincter decreased by 8.3%, 15.6%, 24.3%, and 35.7%, respectively. Additionally, when the external sphincter was injured by 25%, 50%, 75%, and 100%, urethral stress near the external sphincter was reduced by 13.3%, 24.3%, 38.6%, and 46.6%, respectively. Internal sphincter injury primarily affects positions near the internal sphincter and prostate, while external sphincter injury affects the area between the prostate and urethral outlet. These data could facilitate the standardized evaluation of internal and external sphincter dysfunction and lead to novel methods of preoperative assessment for AUS surgery.

A static magnetic field enhances the repair of osteoarthritic cartilage by promoting the migration of stem cells and chondrogenesis.

Objective: To investigate the therapeutic effects of static magnetic field (SMF) and its regulatory mechanism in the repair of osteoarthritic cartilage. Methods: Fourteen-week-old female C57BL/6 mice were randomly divided into the sham operation group and the osteoarthritis (OA) groups with and without SMF application. SMF was applied at 200 â€‹mT for two consecutive weeks. Changes in knee cartilage were examined by histomorphometry, and the chondrogenesis and migration of endogenous stem cells were assessed. The expression of SRY-related protein 9 (SOX9), Collagen type II (COL2), matrix metallopeptidase 13 (MMP13), stromal cell-derived factor 1/C-X-C chemokine receptor type 4 (SDF-1/CXCR4), Piezo1 and other genes was evaluated, and the mechanism of SMF's action was tested using the CXCR4 inhibitor, AMD3100, and Piezo1 siRNA. Results: SMF significantly decreased the OARSI scores after induction of OA. SMF was beneficial to chondrogenesis by elevating SOX9. In the OA mouse model, an increase in MMP13 with a decrease in COL2 led to the destruction of the cartilage extracellular matrix, which was suppressed by SMF. SMF promoted the migration of cartilage-derived stem/progenitor cells and bone marrow-derived mesenchymal stem cells (MSCs). It increased SDF-1 and CXCR4, while the CXCR4 inhibitor significantly suppressed the beneficial effects of SMF. The application of Piezo1 siRNA inhibited the SMF-induced increase of CXCR4. Conclusion: SMF enhanced chondrogenesis and improved cartilage extracellular matrices. It activated the Piezo1-mediated SDF-1/CXCR4 regulatory axis and promoted the migration of endogenous stem cells. Collectively, it attenuated the pathological progression of cartilage destruction in OA mice. The Translational potential of this article: The findings in this study provided convincing evidence that SMF could enhance cartilage repair and improve OA symptoms, suggesting that SMF could have clinical value in the treatment of OA.

Mechanical Loading Promotes the Migration of Endogenous Stem Cells and Chondrogenic Differentiation in a Mouse Model of Osteoarthritis.

Osteoarthritis (OA) is a major health problem, characterized by progressive cartilage degeneration. Previous works have shown that mechanical loading can alleviate OA symptoms by suppressing catabolic activities. This study evaluated whether mechanical loading can enhance anabolic activities by facilitating the recruitment of stem cells for chondrogenesis. We evaluated cartilage degradation in a mouse model of OA through histology with H&E and safranin O staining. We also evaluated the migration and chondrogenic ability of stem cells using in vitro assays, including immunohistochemistry, immunofluorescence, and Western blot analysis. The result showed that the OA mice that received mechanical loading exhibited resilience to cartilage damage. Compared to the OA group, mechanical loading promoted the expression of Piezo1 and the migration of stem cells was promoted via the SDF-1/CXCR4 axis. Also, the chondrogenic differentiation was enhanced by the upregulation of SOX9, a transcription factor important for chondrogenesis. Collectively, the results revealed that mechanical loading facilitated cartilage repair by promoting the migration and chondrogenic differentiation of endogenous stem cells. This study provided new insights into the loading-driven engagement of endogenous stem cells and the enhancement of anabolic responses for the treatment of OA.

Three-dimensional measurement and analysis of morphological parameters of the uterus in infertile women.

Background: To determine differences in endometrial cavity anteroposterior diameter, thickness, volume, and diameter lines of uterine body and thickness, and volume of upper, middle, and lower regions of the endometrium in infertile women using a new method for three-dimensional (3D) reconstruction based on two-dimensional (2D) ultrasound images. Methods: This retrospective cross-sectional study included a total of 81 infertile women, who underwent 2D ultrasound standard examination. We created 3D models of the uterine body, endometrial cavity, and endometrium based on 2D ultrasound images. The parameters that were measured and analyzed in a 3D plane included volume and diameter lines of endometrial cavity, surface area, thickness, volume, and diameter lines of uterine body, and surface area, thickness, and volume of upper, middle, and lower region of the endometrium. These parameters were used for comparisons between normal and arcuate uterus, between non-pregnant and pregnant infertile women, and between nulliparous and multiparous infertile women. The differences between the different regions of the endometrium and the correlations between age and the parameters were also determined in this study. Results: Endometrial cavity length, and middle and lower regions of the endometrial volume in the normal uterus were 39.63±7.61 mm, 1,307.92±1,034.40 mm3, and 653.98±460.41 mm3, respectively. For arcuate uterus, these parameters were 32.96±4.69 mm, 539.89±298.94 mm3, and 347.90±129.61 mm3, respectively. The parameters were significantly higher in normal uterus compared with arcuate uterus (P=0.000, 0.001, and 0.006, respectively). Upper, middle, and lower regions of endometrial thickness in normal uterus were 7.79±3.26, 8.18±3.33, and 6.41±2.60 mm, respectively. Both upper and middle regions of endometrial thickness were significantly greater than the lower regions of endometrial thickness with P=0.009 and P=0.001, respectively. Correlation analysis revealed that age positively correlated with volume of upper endometrial regions (r=0.274, P=0.028). Conclusions: This study provides references for the volume and thickness of the endometrium in the different anatomical regions of normal and arcuate uterus. Age mainly affects the upper region of the endometrium. The 3D measurement provides a precise way to quantify the morphological parameters of gynecological diseases.

Knee Loading Enhances the Migration of Adipose-Derived Stem Cells to the Osteoarthritic Sites Through the SDF-1/CXCR4 Regulatory Axis.

Osteoarthritis (OA) is a whole joint disorder that is characterized by cartilage damage and abnormal remodeling of subchondral bone. Injecting adipose-derived stem cells (ASCs) into the knee joint cavity can assist in repairing osteoarthritic joints, but their ability to migrate to the damaged site is limited. Our previous studies have shown that knee loading can improve the symptoms of OA, but the effect and mechanism of knee loading on the migration of ASCs in OA remain unclear. We employed a mouse model of OA in the knee and applied knee loading (1 N at 5 Hz for 6 min/day for 2 weeks) after the intra-articular injection of ASCs. The cartilage and subchondral bone repair were assessed by histopathological analysis. Immunofluorescence assays were also used to analyze the migration of ASCs. Using cell cultures, we evaluated the migration of ASCs using the transwell migration and wound healing assays. In vivo experiments showed that knee loading promoted the migration of ASCs, increased the local SDF-1 level, and accelerated the repair of the OA-damaged sites. Mechanistically, the observed effects were blocked by the SDF-1/CXCR4 inhibitor. The in vitro results further revealed that knee loading promoted the migration of ASCs and the inhibition of SDF-1/CXCR4 significantly suppressed the beneficial loading effect. The results herein suggested that the migration of ASCs was enhanced by knee loading through the SDF-1/CXCR4 regulatory axis, and mechanical loading promoted the joint-protective effect of ASCs.

Mechanical loading mitigates osteoarthritis symptoms by regulating the inflammatory microenvironment in a mouse model.

Osteoarthritis (OA) is one of the most common chronic diseases, in which inflammatory responses in the articular cavity induce chondrocyte apoptosis and cartilage degeneration. While mechanical loading is reported to mitigate synovial inflammation, the mechanism and pathways for the loading-driven improvement of OA symptoms remain unclear. In this study, we evaluated the loading effects on M1/M2 polarization of synovial macrophages via performing histology, cytology, and molecular analyses. In the OA group, the cell layer of the synovial lining was enlarged with an increase in cell density. Also, M1 macrophages were polarized and proinflammatory cytokines were increased. In contrast, in the OA group with mechanical loading, cartilage degradation was reduced and synovial inflammation was alleviated. Notably, the M1 macrophages were diminished by mechanical loading, while M2 macrophages were increased. Furthermore, mechanical loading decreased the levels of proinflammatory cytokines, such as interleukin-1 beta and tumor necrosis factor-α, and suppressed PI3K/AKT/NF-κB signaling. Taken together, this study demonstrates that mechanical loading changes the ratio of M1 and M2 macrophages via regulation of PI3K/AKT/NF-κB signaling and provides cartilage protection in the mouse OA model.

Loading-driven PI3K/Akt signaling and erythropoiesis enhanced angiogenesis and osteogenesis in a postmenopausal osteoporosis mouse model.

Bone vasculature influences osteogenesis and haematopoiesis in the bone microenviroment. Mechanical loading has been shown to stimulate the formation of osteogenesis-related type H vessels in an ovariectomy (OVX)-induced osteoporosis mouse model. To determine the loading-driven mechanism of angiogenesis and the formation of type H vessels in bone, we evaluated the roles of PI3K/Akt signaling and erythropoiesis in the bone marrow. The daily application of mechanical loading (1 N at 5 Hz for 6 min/day) for 2 weeks on OVX mice inhibited osteoclast activity, associated with an increase in the number of osteoblasts and trabecular volume ratio. Mechanical loading enhanced bone vasculature and vessel formation, as well as PI3K/Akt phosphorylation and erythropoiesis in the bone marrow. Notably, LY294002, an inhibitor of PI3K signaling, blocked the tube formation by endothelial progenitor cells, as well as their migration and wound healing. The conditioned medium, derived from erythroblasts, also promoted the function of HUVECs with elevated levels of VEGF, CD31, and Emcn. Collectively, this study demonstrates that mechanical loading prevents osteoporotic bone loss by promoting angiogenesis and type H vessel formation. This load-driven preventing effect is in part mediated by PI3K/Akt signaling and erythropoiesis in the bone marrow.

Three-dimensional measurement and analysis of benign prostatic hyperplasia.

BACKGROUND: The volume and thickness of intravesical prostatic protrusion and other characteristics of benign prostatic hyperplasia have not been investigated. We determine the effects of age and prostate volume on anatomical features of benign prostatic hyperplasia using three-dimensional measurement in this study. METHODS: This retrospective study included a total of 98 patients with benign prostatic hyperplasia. Three-dimensional models of prostate, central gland, peripheral zone, intravesical prostatic protrusion, prostatic urethra and bladder were reconstructed according to pelvic T2-weighted magnetic resonance imaging of these patients. The models were used to measure the intravesical prostatic protrusion volume, intravesical prostatic protrusion thickness, intravesical prostatic protrusion index, intravesical prostatic protrusion, prostate volume, peripheral zone volume, peripheral zone thickness, peripheral zone index, prostatic urethra thickness, the angle and distance of distal prostatic urethra with regard to coronal plane and sagittal plane and so on. RESULTS: Intravesical prostatic protrusion volume, intravesical prostatic protrusion thickness and peripheral zone volume of prostate volume >80 mL group were significantly higher than these in prostate volume <80 mL group (P<0.001, 0.01, 0.01, respectively). These parameters significantly increased with age (P<0.001, 0.01, 0.05, respectively). Peripheral zone index was significantly lower of prostate volume >80 mL group than these in prostate volume <80 mL group (P<0.05). Peripheral zone index significantly decreased with age (P<0.01). Intravesical prostatic protrusion index had no significant difference in all age groups. Peripheral zone thickness and prostatic urethra thickness had no significant difference in all groups. The distance and angle of distal prostatic urethra prostatic urethra with regard to coronal plane were significantly higher than these with regard to sagittal plane (both P<0.001). CONCLUSIONS: The rearward slope of the prostatic urethra is greater than the left or right offset during the process of benign prostatic hyperplasia. Three-dimensional measurement provides good supports for further clinical and scientific research.

The distribution pattern of periprostatic neurovascular bundles examined with successive celloidin slices.

BACKGROUND: Although several distribution patterns of periprostatic neurovascular bundles have been proposed, variant dissection technique based on these patterns still confused surgeons. The aim of this study was to describe the periprostatic neurovascular bundles and their relationship with the fascicles around prostate and provide the accurate morphologic knowledge of periprostatic tissue for prostate operation. METHODS: The pelvic viscera were obtained from 26 adult male cadavers. They were embedded in celloidin and cut into successive slices. The slices were explored with anatomic microscopy. 3-Dimensional reconstruction was achieved with celloidin sections and series software. RESULTS: The prostatic capsule which surrounded the dorsal, bilateral aspect of the prostate was attached ventrally to anterior fibrous muscular stroma (AFMS). The lower part of the striated sphincter completely embraced the urethral; the upper part of this muscle covered the lower ventral surface of prostate. The upper ventral surface of prostate is covered by the circular muscle of detrusor. The levator fascia and the capsule adhered on the most convex region of the lateral prostate, but separated on the other region. The pelvic neurovascular bundles (PNVB) divided into the anterior and posterior divisions. The anterior division continued as dorsal vascular complex (DVC). The distal part of DVC entered into penile hilum. The posterior division continued as neurovascular bundles, and then as the cavernous supply (CS). The distal part of CS joined into pudendal neurovascular bundles. CONCLUSIONS: The capsule and AFMS formed a pocket like complex. There were anterior and posterior neurovascular approaches from PNVB to penile hilum.

Mechanical loading stimulates bone angiogenesis through enhancing type H vessel formation and downregulating exosomal miR-214-3p from bone marrow-derived mesenchymal stem cells.

Exosomes are important transporters of miRNAs, which play varying roles in the healing of the bone fracture. Angiogenesis is one of such critical events in bone healing, and we previously reported the stimulatory effect of mechanical loading in vessel remodeling. Focusing on type H vessels and exosomal miR-214-3p, this study examined the mechanism of loading-driven angiogenesis. MiRNA sequencing and qRT-PCR revealed that miR-214-3p was increased in the exosomes of the bone-losing ovariectomized (OVX) mice, while it was significantly decreased by knee loading. Furthermore, compared to the OVX group, exosomes, derived from the loading group, promoted the angiogenesis of endothelial cells. In contrast, exosomes, which were transfected with miR-214-3p, decreased the angiogenic potential. Notably, knee loading significantly improved the microvascular volume, type H vessel formation, and bone mineral density and contents, as well as BV/TV, Tb.Th, Tb.N, and Tb.Sp. In cell cultures, the overexpression of miR-214-3p in endothelial cells reduced the tube formation and cell migration. Collectively, this study demonstrates that knee loading promotes angiogenesis by enhancing the formation of type H vessels and downregulating exosomal miR-214-3p.

[Finite element analysis of male lower urinary tract based on the collodion slice images].

Males typically have high rates of morbidity of primary bladder neck obstruction, while the existing urodynamic examination is invasive and more likely to cause false diagnosis. To build a non-invasive biomechanical detecting system for the male lower urinary tract, a finite element model for male lower urinary tract based on the collodion slice images of normal male lower urinary tract was constructed, and the fluid-structure interaction of the lower urinary tract was simulated based on the real urination environment. The finite element model of the lower urinary tract was validated by comparing the clinical experiment data with the simulation result. The stress, flow rate and deformation of the lower urinary tract were analyzed, and the results showed that the Von Mises stress and the wall shear stress at the membrane sphincter in the normal male lower urinary tract model reached a peak, and there was nearly 1 s delay than in the bladder pressure, which helped to validate the model. This paper lays a foundation for further research on the urodynamic response mechanism of the bladder pressure and flow rate of the lower urinary tract obstruction model, which can provide a theoretical basis for the research of non-invasive biomechanical detecting system.

Effects of knee loading on obesity-related non-alcoholic fatty liver disease in an ovariectomized mouse model with high-fat diet.

AIM: Hormonal and nutritional disorders are the main causes of obesity and non-alcoholic fatty liver disease, especially in the elderly and in postmenopausal women. Although physical activity might alleviate these disorders, the elderly may often have difficulty in carrying out physical exercise. The purpose of this study was to investigate the therapeutic effect of knee loading, a new form of physical stimulation, on the symptoms of obesity and fatty liver. METHODS: Using ovariectomized mice fed a high-fat diet, we evaluated the effect of knee loading that applies gentle cyclic loads to the knee. Female C57BL/6 mice were divided into five groups: control (SCD), high-fat diet (HF), HF with loading (HF + L), HF with ovariectomy (HF + OVX), and HF + OVX with loading (HF + OVX + L). Except for SCD, mice underwent sham operation or ovariectomy and were maintained on HF diet. After 6 weeks, the mice in the HF + L and HF + OVX + L groups were treated with knee loading for 6 weeks. RESULTS: Compared to the obesity groups (HF and HF + OVX), knee loading significantly decreased a gain in body weight, liver weight, and white adipose tissue (all P < 0.01). It also reduced the lipid level in the serum (P < 0.01) and histological severity of hepatic steatosis (P < 0.01). Furthermore, knee loading downregulated biomarkers related to endoplasmic reticulum (ER) stress (GRP78, p-eIF2α, and ATF4) and altered biomarkers in autophagy (LC3 and p62). CONCLUSIONS: Knee loading suppressed obesity-associated metabolic alterations and hepatic steatosis. These effects with knee loading might be associated with suppression of ER stress and promotion of autophagy.

Effect of Methionine Restriction on Bone Density and NK Cell Activity.

Methionine restriction (MR) is proven to increase the lifespan; and it also affects the bone density and the innate immune system. The aim of this study is to explore the effect of methionine restriction on bone density and natural killer (NK) cells. C57BL/6J mice were subjected to either basal diet (BD, containing 0.80% methionine) or methionine-restricted diet (containing 0.14% methionine). Mice with MR diet displayed reduced bone mass and decrease in the cytotoxicity of NK from the spleen, compared to BD animals. Also, mice with MR diet had an inferior body weight (P < 0.05) and higher plasma levels of adiponectin and FGF21 (P < 0.05) but lower concentrations of leptin and IGF-1 (P < 0.05). Overall, the investigation shows that methionine affects bone density and NK cell cytotoxicity.

High-Methionine Diet Attenuates Severity of Arthritis and Modulates IGF-I Related Gene Expressions in an Adjuvant Arthritis Rats Model.

Rheumatoid arthritis, a synthesized form of adjuvant arthritis exhibited throughout many animal species, inhibits liver function and circulation of IGF-I and contributes to the degradation of skeletal muscle mass. One of the primary goals of the present study is determining whether a high-Methionine (high-Met) diet is capable of reducing the adverse effects of arthritis, namely, loss of body mass. Following adjuvant injection, forty arthritic rats were randomly assigned to either a control group with a basal diet or a high-Met group with the same basal diet + 0.5% Methionine. After 14 days all rats were terminated. The high-Met group exhibited an increase in body weight and food intake in comparison with the control group (P < 0.05). High-Met diet debilitated arthritis-induced surges in the gastrocnemius in both atrogin-1 and the MuRF1 expressions; however, it was observed to have little to no effect on atrogin-1 and MuRF1 gene expression in soleus. At the same time, high-Met diet rats experienced a rise in IGF-I, with lowering of IGFBP-3 gene expression in the gastrocnemius and the soleus. These data suggest that arthritis severity can be partly attenuated by high-Met diet.

[Effects of chemerin on the proliferation of 3T3-L1 adipocytes].

OBJECTIVE: To explore the effects of chemerin on the proliferation of 333-Li preadipocytes and elucidate its possible mechanism. METHODS: Recombinant lentivirus-mediated silencing or over-expression of chemerin gene were constructed in 3T3-L1 cells. The proliferation of 3T3-L1 cells was examined by the assays of methyl thiazolyl tetrazolium (MTT) and 5-bromo-2-deoxyuridine (BrdU) incorporation. The expressions of chemerin, ERK1, ERK2 and cyclinD1 in 3T3-L1 cells were detected by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The protein levels of chemerin, ERK1/2 and p-ERK1/2 were detected by Western blot. RESULTS: MT and BrdU results showed that absorbance (A) value and BrdU incorporation increased in chemerin over-expression group compared with over-expression control group [1. 09 ± 0. 08 vs 0. 90 ± 0. 09, (126. 5 ± 14. 6)% vs (100. 0 ± 7. 0) %, both P <0. 05]. The increase of A value and BrdU incorporation in chemerin over-expression group could be inhibited by PD98059, a blocker of ERK1/2 signal pathway. A value (0. 91 ± 0. 13) and BrdU incorporation [(104. 3 ± 10. 7)%] decreased compared with over-expression group (P <0. 05). They also decreased in chemerin gene silencing group compared to silencing control group [ 0. 72 ± 0. 11 vs 0. 90 ± 0. 09, (77. 6 ± 11. 8) % vs ( 99. 7 ± 6. 3) %, both P < 0. 05]. Quantitative real-time PCR revealed that over-expression group had a higher expression in chemerin, ERK, ERK2 and cyclinDl than over-expression control group (2. 77 ± 0. 31 vs 1. 01 ± 0. 12, 1. 39 ± 0. 19 vs 0. 76 ± 0. 30, 1. 46 ± 0. 14 vs 0. 88 ± 0. 14, 1. 44 ± 0. 17 vs 1. 03 0. 15, all P <0. 05). Chemerin gene silencing group had lower expressions of chemerin, ERKI, ERK2 and cyclinD1 than silencing control group (0. 35 ± 0. 12 vs 1. 25 ± 0. 31, 0. 64 ± 0. 15 vs 1. 03 ± 0. 14, 0. 56 ± 0. 10 vs 1. 06 ± 0. 29, 0. 66 ± 0. 13 vs 1. 09 0. 19, all P <0. 05). Western blot showed that the expressiond of chemerin, ERK1/2 and p-ERK1/2 increased in chemerin over-expression group versus over-expression control group (2. 18 ± 0. 32 vs 1. 18 ± 0. 14, 1. 37 ± 0. 05 vs 0. 97 ± 0. 12, 1. 06 ± 0. 09 vs 0. 56 ± 0. 08, all P < 0. 05). The expressiond of chemerin, ERK1/2 and p-ERK1/2 decreased in chemerin gene silencing group versus silencing control group (1. 00 ± 0. 07 vs 1. 40 ± 0. 17, 0. 87 ± 0. 15 vs 1. 20 ± 0. 12, 0. 49 ± 0. 07 vs 0. 91 ± 0. 11, all P < 0. 05). CONCLUSIONS: Chemerin may promote the proliferation of 3T3-L1 cells. And it may be achieved via an up-regulated expression of ERK1/2.

Muscular structure at the male bladder outlet examined with successive celloidin-embedded slices.

OBJECTIVE: To re-examine the detailed anatomy of the muscular system at the bladder neck and proximal urethra in the male and to explore its function in urinary continence and micturition further. METHODS: The pelvic organs, including bladder, prostate, and rectum, were obtained from 20 formalin-fixed adult male cadavers and were removed from the pelvic cavity and embedded in celloidin in their entirety. The embedded block was cut into successive slices with an immersing-alcohol microtome. RESULTS: Circular muscle fibers of the detrusor at the bladder outlet consist of the anterior downward-projected circular muscle fibers of the bladder outlet (ADPC), the bilateral accumulated circular fibers, and the posterior circular fibers of the bladder outlet. Together, these fibers concentrically surround the internal urethral orifice and trigone muscle. The lower part of the ADPC surrounds the ventral surface of the proximal urethra. Longitudinal muscle fibers are radially inserted into the circular muscle around the internal urethral orifice. Numerous fibers from the ventral longitudinal muscle are inserted into the lower part of the ADPC. The upper part of the trigone muscle exists in bladder cavity; the lower part extends into the proximal urethra to surround the posterior and posterolateral surface of the urethra. CONCLUSION: The ADPC and the upward extension of the rhabdosphincter comprise the anterior fibromuscular stroma. The circular muscle of the bladder outlet may be responsible for closure; the longitudinal muscle of the bladder outlet may be responsible for opening of the internal urethral orifice and proximal urethra.