A novel method to establish a rat ED model using internal iliac artery ligation combined with hyperlipidemia.

OBJECTIVE: To investigate a novel method, namely using bilateral internal iliac artery ligation combined with a high-fat diet (BCH), for establishing a rat model of erectile dysfunction (ED) that, compared to classical approaches, more closely mimics the chronic pathophysiology of human ED after acute ischemic insult. MATERIALS AND METHODS: Forty 4-month-old male Sprague Dawley rats were randomly placed into five groups (n = 8 per group): normal control (NC), bilateral internal iliac artery ligation (BIIAL), high-fat diet (HFD), BCH, and mock surgery (MS). All rats were induced for 12 weeks. Copulatory behavior, intracavernosal pressure (ICP), ICP/mean arterial pressure, hematoxylin-eosin staining, Masson's trichrome staining, serum lipid levels, and endothelial and neuronal nitric oxide synthase immunohistochemical staining of the cavernous smooth muscle and endothelium were assessed. Data were analyzed by SAS 8.0 for Windows. RESULTS: Serum total cholesterol and triglyceride levels were significantly higher in the HFD and BCH groups than the NC and MS groups. High density lipoprotein levels were significantly lower in the HFD and BCH groups than the NC and MS groups. The ICP values and mount and intromission numbers were significantly lower in the BIIAL, HFD, and BCH groups than in the NC and MS groups. ICP was significantly lower in the BCH group than in the BIIAL and HFD groups. Cavernous smooth muscle and endothelial damage increased in the HFD and BCH groups. Cavernous smooth muscle to collagen ratio, nNOS and eNOS staining decreased significantly in the BIIAL, HFD, and BCH groups compared to the NC and MS groups. CONCLUSIONS: The novel BCH model mimics the chronic pathophysiology of ED in humans and avoids the drawbacks of traditional ED models

Metabolomics Analysis of Seminal Plasma in Infertile Males with Kidney-Yang Deficiency: A Preliminary Study

Traditional Chinese medicine (TCM) is an important treatment for male infertility, and its application to therapy is dependent on differentiation of TCM syndromes. This study aims to investigate the changes in metabolites and metabolic pathways in infertile males with Kidney-Yang Deficiency syndrome (KYDS) via metabolomics approaches. Seminal plasma samples were collected from 18 infertile males with KYDS and 18 fertile males. Liquid chromatography and mass spectrometry were used to characterize metabolomics profiles. Principal component analysis (PCA), partial least squares-discriminate analysis (PLS-DA), and pathway analysis were used for pattern recognition and metabolite identification. PCA and PLS-DA results differentiated the two groups of patients. Forty-one discriminating metabolites (18 in positive mode and 23 in negative mode) were identified. Seven metabolites were related to five potential metabolic pathways associated with biosynthesis and metabolism of aromatic amino acids, tricarboxylic acid cycle, and sphingolipid metabolism. The changes in metabolic pathways may play an important role in the origin of KYDS-associated male infertility. Metabolomics analysis of seminal plasma may be used to differentiate TCM syndromes of infertile males, but further research must be conducted

Commonly used mesenchymal stem cell markers and tracking labels: Limitations and challenges.

Early observations that cultured mesenchymal stem cells (MSCs) could be induced to exhibit certain characteristics of osteocytes and chondrocytes led to the proposal that they could be transplanted for tissue repair through cellular differentiation. Therefore, many subsequent preclinical studies with transplanted MSCs have strived to demonstrate that cellular differentiation was the underlying mechanism for the therapeutic effect. These studies generally followed the minimal criteria set by The International Society for Cellular Therapy in assuring MSC identity by using CD70, CD90, and CD105 as positive markers and CD34 as a negative marker. However, the three positive markers are co-expressed in a wide variety of cells, and therefore, even when used in combination, they are certainly incapable of identifying MSCs in vivo. Another frequently used MSC marker, Stro-1, has been shown to be an endothelial antigen and whether it can identify MSCs in vivo remains unknown. On the other hand, the proposed negative marker CD34 has increasingly been shown to be expressed in native MSCs, such as in the adipose tissue. It has also helped establish that MSCs are likely vascular stem cells (VSCs) that reside in the capillaries and in the adventitia of larger blood vessels. These cells do not express CD31, CD104b, or α-SMA, and therefore are designated as CD34+CD31-CD140b-SMA-. Many preclinical MSC transplantation studies have also attempted to demonstrate cellular differentiation by using labeled MSCs. However, all commonly used labels have shortcomings that often complicate data interpretation. The β-gal (LacZ) gene as a label is problematic because many mammalian tissues have endogenous β-gal activities. The GFP gene is similarly problematic because many mammalian tissues are endogenously fluorescent. The cell membrane label DiI can be adsorbed by host cells, and nuclear stains Hoechst dyes and DAPI can be transferred to host cells. Thymidine analog BrdU is associated with loss of cellular protein antigenicity due to harsh histological conditions. Newer thymidine analog EdU is easier to detect by chemical reaction to azide-conjugated Alexa fluors, but certain bone marrow cells are reactive to these fluors in the absence of EdU. These caveats need to be taken into consideration when designing or interpreting MSC transplantation experiments

Commonly used mesenchymal stem cell markers and tracking labels: Limitations and challenges

Early observations that cultured mesenchymal stem cells (MSCs) could be induced to exhibit certain characteristics of osteocytes and chondrocytes led to the proposal that they could be transplanted for tissue repair through cellular differentiation. Therefore, many subsequent preclinical studies with transplanted MSCs have strived to demonstrate that cellular differentiation was the underlying mechanism for the therapeutic effect. These studies generally followed the minimal criteria set by The International Society for Cellular Therapy in assuring MSC identity by using CD70, CD90, and CD105 as positive markers and CD34 as a negative marker. However, the three positive markers are co-expressed in a wide variety of cells, and therefore, even when used in combination, they are certainly incapable of identifying MSCs in vivo. Another frequently used MSC marker, Stro-1, has been shown to be an endothelial antigen and whether it can identify MSCs in vivo remains unknown. On the other hand, the proposed negative marker CD34 has increasingly been shown to be expressed in native MSCs, such as in the adipose tissue. It has also helped establish that MSCs are likely vascular stem cells (VSCs) that reside in the capillaries and in the adventitia of larger blood vessels. These cells do not express CD31, CD104b, or α-SMA, and therefore are designated as CD34+CD31-CD140b-SMA-. Many preclinical MSC transplantation studies have also attempted to demonstrate cellular differentiation by using labeled MSCs. However, all commonly used labels have shortcomings that often complicate data interpretation. The ß-gal (LacZ) gene as a label is problematic because many mammalian tissues have endogenous ß-gal activities. The GFP gene is similarly problematic because many mammalian tissues are endogenously fluorescent. The cell membrane label DiI can be adsorbed by host cells, and nuclear stains Hoechst dyes and DAPI can be transferred to host cells. Thymidine analog BrdU is associated with loss of cellular protein antigenicity due to harsh histological conditions. Newer thymidine analog EdU is easier to detect by chemical reaction to azide-conjugated Alexa fluors, but certain bone marrow cells are reactive to these fluors in the absence of EdU. These caveats need to be taken into consideration when designing or interpreting MSC transplantation experiments

The histological analysis of rat corpus cavernosum in NC, BIIAL, HFD, BCH and MS groups after 12 weeks.

<p>Sections were made transversal to the penile axis. HE staining showed the changes in the cavernous smooth muscle, collagen and endothelium in rat corpus cavernosum in the different groups. (A) and (B) show the appropriate appearance of the cavernous smooth muscle and intact endothelium of the rat cavernosum in the NC group. (C) and (D) show decreased cavernous smooth muscle content and moderately damaged endothelium in the BIIAL group. (E) and (F) show increased cavernous smooth muscle content and a thickened endothelium in the HFD group. (G) and (H) show decreased cavernous smooth muscle content and severely damaged endothelium in the BCH group. (I) and (J) show the similar normal appearance of the cavernous smooth muscle and intact endothelium of the rat cavernosum in the MS and NC groups. A, C, E, G, original magnification ×100. B, D, F, H, original magnification ×200.</p

The eNOS immunohistochemical staining of rat corpus cavernosum in NC, BIIAL, HFD, BCH and MS groups after 12 weeks.

<p>Sections were made transversal to the penile axis. (A) Normal eNOS expression was observed in the NC group. (B) Mildly decreased eNOS expression was observed in the BIIAL group. (C) Severely decreased eNOS expression was observed in the HFD group. (D) Significantly decreased eNOS expression was observed in the BCH group. (E) Similar eNOS expression was observed in the MS and NC groups.</p

nNOS and eNOS positive rates.

<p>* <i>p</i><0.05 vs. NC group;</p><p># <i>p</i><0.05 vs. BCH group.</p

Changes of body weight of 5 groups.

<p>* <i>p</i><0.05 vs. other groups.</p

The ICP levels of NC, BIIAL, HFD, BCH and MS groups after 12 weeks.

<p>(A) In the NC group, the ICP in the rat corpus cavernosum increased greatly during a normal erection induced by apomorphine and lasted for a relatively long period. (B) In the BIIAL group, the ICP in the rat corpus cavernosum showed a low increase and short duration during an invalid erection. (C) In the HFD group, the ICP in the rat corpus cavernosum showed a low peak but a relatively long duration during an invalid erection. (D) In the BCH group, the ICP in the rat corpus cavernosum showed an extremely slow increase and minor peak, and almost no duration for the erection was identified. (E) In the MS group, the ICP in the rat corpus cavernosum showed a rapid increase and a long duration similar to the NC group.</p

Mount number, intromission number, increase in ICP and ICP/MAP and duration.

<p>* <i>p</i><0.05 vs. NC group;</p><p># <i>p</i><0.05 vs. BCH group.</p