Absence of PD-L1 signaling hinders macrophage defense against Mycobacterium tuberculosis via upregulating STAT3/IL-6 pathway.

The blockade of programmed death-ligand 1 (PD-L1) pathway has been clinically used in cancer immunotherapy, while its effects on infectious diseases remain elusive. Roles of PD-L1 signaling in the macrophage-mediated innate immune defense against M.tb is unclear. In this study, the outcomes of tuberculosis (TB) in wild-type (WT) mice treated with anti-PD-1/PD-L1 therapy and macrophage-specific Pdl1-knockout (Pdl1ΔΜΦ) mice were compared. Treatment with anti-PD-L1 or anti-PD-1 benefited protection against M.tb infection in WT mice, while Pdl1ΔΜΦ mice exhibited the increased susceptibility to M.tb infection. Mechanistically, the absence of PD-L1 signaling impaired M.tb killing by macrophages. Furthermore, elevated STAT3 activation was found in PD-L1-deficient macrophages, leading to increased interleukin (IL)-6 production and reduced inducible nitric oxide synthase (iNOS) expression. Inhibiting STAT3 phosphorylation partially impeded the increase in IL-6 production and restored iNOS expression in these PD-L1-deficient cells. These findings provide valuable insights into the complexity and mechanisms underlying anti-PD-L1 therapy in the context of tuberculosis.

Aberrant branches of the subclavian artery and their relationship with the phrenic nerve and the brachial plexus / Ramas aberrantes de la arteria subclavia y su relación con el nervio frénico y el plexo braquial

SUMMARY: Variations in subclavian artery branches are relatively common and may impact surgical procedures and effects. During educational dissection of a male cadaver, we encountered an extremely rare variation of the right subclavian artery branches. The internal thoracic artery, the thyrocervical trunk, and the costocervical trunk arose from the third part of the right subclavian artery. In addition, the phrenic nerve displaced remarkably laterally by the thyrocervical trunk, and the course of the costocervical trunk was between the upper trunk and the middle trunk of the brachial plexus. These variations may pose a potential risk for nerve compression and increase the risk of arterial and nerve puncture. This case report would bring attention to the possibility of other similar cases, and early detection of these variations through diagnostic interventions is helpful to reduce postoperative complications.

RESUMEN: Las variaciones en las ramas de la arteria subclavia son relativamente comunes y pueden afectar los procedimientos y efectos quirúrgicos. Durante la disección educativa de un cadáver masculino, encontramos una variación extremadamente rara de las ramas de la arteria subclavia derecha. La arteria torácica interna, el tronco tirocervical y el tronco costocervical nacían de la tercera parte de la arteria subclavia derecha. Además, el nervio frénico se desplazaba lateralmente por el tronco tirocervical, y el trayecto del tronco costocervical se encontraba entre el tronco superior y el tronco medio del plexo braquial. Estas variaciones pueden suponer un riesgo potencial de compresión nerviosa y aumentar el riesgo de punción arterial y nerviosa. Este reporte de caso llamaría la atención sobre la posibilidad de otros casos similares, y la detección temprana de estas variaciones a través de diagnósticos es útil para reducir las complicaciones postoperatorias.

The role of Snf5 in the osteogenic differentiation potential during replicative senescence of rat mesenchymal stromal cells.

The osteogenic capacities of bone marrow-derived stromal cells (BMSCs) diminish during replicative senescence, and these changes affect the success of therapeutic application of BMSCs. In this study, we sought to explore the molecular mechanisms underlying the osteogenic differentiation capacities that occur during replicative senescence. It is well known that Oct4 is a key transcription factor essential for maintaining differentiation capacities of the stem cells. In this study, we found that BMSCs at passage 6 (replicative senescent BMSCs) showed marked decreases in the osteogenic differentiation potential and the level of Oct4. These were accompanied by reduced levels of Snf5 and histone H3 lysine-4 trimethylation (H3K4me3) in the Oct4 promoter. In BMSCs at passage 2, knockdown of Snf5 diminished expression of Oct4 and disrupted the up-regulation of alkaline phosphatase (ALP) and runt-related transcription factor 2 (Runx2) after osteogenic differentiation induction, which was accompanied by a reduction in Snf5 and H3K4me3 binding to the Oct4 promoter. These findings indicate that the decreased level of Snf5 binding to the promoter region of the Oct4 gene down-regulated the expression of Oct4, which may be the mechanism underlying the decline in osteogenic capacities in replicative senescent BMSCs.

A rare variation of the testicular blood supply by the deferential artery / Rara variación del suministro de sangre testicular por la arteria del conducto deferente

Any compression of testicular arteries may lead to loss of spermatogenesis and gonadal hormone production, existence of the variational arteries is accountable in cases of vasoligation, orchidopexy and other surgical approach on them. Anomalies of the testicular blood arteries were observed during dissection of the pelvic cavity in a 68-year-old male cadaver. This report describes a very rare case of lack of testicular arteries. For the blood supply to the testis, thick deferential arteries form some vascular winding and loops and course accompanied by deferent duct to the testis. This case report would serve as ray of light for knowledge of the possible variations of the testicular arteries during surgical procedures.

Cualquier compresión de las arterias testiculares puede conducir a la pérdida de la espermatogénesis y la producción de hormonas gonadales. La existencia de variaciones en las arterias testiculares es relevante en los casos de vasectomía, orquidopexia y otros tipos de abordaje quirúrgico. Se observaron anomalías de las arterias testiculares durante la disección de la cavidad pélvica de un cadáver de sexo masculino de 68 años de edad. En este trabajo se describe un caso muy poco frecuente de ausencia de arterias testiculares. Para el suministro sanguíneo del testículo, se encontraron arterias deferentes gruesas que producen sinuosidad y tortuosidad vascular junto al conducto deferente en los testículos. Este caso podría ser útil para el conocimiento de las posibles variaciones de las arterias testiculares durante los procedimientos quirúrgicos.

p53 regulates autophagic activity in senescent rat mesenchymal stromal cells.

The tumor suppressor protein p53 is an important player in the regulation of cell senescence, its functions are largely carried out by modulating its downstream genes. Emerging evidence has suggested that senescence and autophagy appear to be regulated by overlapping signaling pathways. Furthermore, autophagy markers have been observed in senescent cells. In this study, we sought to explore the effects of the expression pattern and function of p53 on the activity of autophagy and replicative senescence in bone marrow derived mesenchymal stromal cells (BMSCs). We found that more than 85% of BMSCs stained positive for SA-ß-gal at passage 6 (senescent BMSCs) with increased expressions of senescence related genes (p16(ink4a) and p21(waf1)). These results were accompanied by the up-regulation of p53, down-regulation of mammalian target of rapamycin (mTOR) and phosphorylation of Rb. Senescent BMSCs displayed an increased monodansylcadaverine (MDC) staining and autophagy related genes (LC3 and atg12) level compared with BMSCs at passage 2. Knockdown of p53 alleviated the senescent state and reduced autophagic activity during the progression of BMSC senescence, which was accompanied by significantly up-regulated levels of mTOR and phosphorylation of Rb. These results demonstrate that autophagy increases when BMSCs enter the replicative senescence state, and p53 contributes a crucial role in the up-regulation of autophagy in this state.

Inhibition of autophagy alleviates the senescent state of rat mesenchymal stem cells during long-term culture.

Following a limited number of cell divisions, mesenchymal stem cells (MSCs) undergo senescence, and these senescent cells maintain metabolic modification and remain viable for long periods. Autophagy, an intracellular bulk degradation process, provides a survival effect for cells under stress. In this study, the effect of autophagy on senescent MSCs was analyzed. Following serial passaging, rat MSCs underwent replicative senescence, characterized by positive staining for senescence-associated ß-galactosidase (SA-ß-gal), and increased expression levels of p16 and p21. During MSC senescence, the levels of autophagic activity were increased, a greater number of autophagic vacuoles were observed in senescent MSCs by transmission electron microscopy, acridine orange staining was elevated and the expression levels of autophagy­related proteins (microtubule­associated protein 1A/1B­light chain 3-II, Atg7 and Atg12) were increased. The role of autophagy in MSC senescence was further investigated through pharmacological inhibition of autophagy with bafilomycin A1 and 3-methyladenine. Inhibition of autophagy by pharmacological means reduced the rate of positive staining for SA-ß-gal and the expression levels of senescence­related proteins. In conclusion, these findings suggest that autophagy is activated during senescence and the autophagic activity may be a requirement for maintaining the senescent state of MSCs.

HPV16 infection regulates RASSF1A transcription mediated by p53.

Human papillomavirus (HPV) 16 infection and RASSF1A expression play important roles in tumor development and progression. However, the precise mechanisms underlying their concerted function in the development of reproductive system tumors still remain to be elucidated. In the present study, we showed that HPV16-E6 selectively upregulates RASSF1A expression via degradation of p53, which interacts with the RASSF1A promoter and regulates apoptosis. Overexpression of p53 triggered a decrease in endogenous RASSF1A in SiHa cells, accompanied by apoptosis. Similarly, knockdown of endogenous HPV16-E6 in SiHa cells with RNA interference (RNAi) led to downregulation of RASSF1A mediated by p53 and the subsequent induction of apoptosis. These findings collectively suggest that HPV16 infection regulates p53-mediated RASSF1A expression and suppresses apoptosis. Moreover, RASSF1A may form an element of the negative autoregulatory feedback loops that act on the HPV16 response and are involved in p53-dependent apoptosis. Our results provide novel insights into the cellular mechanism of tumor development, and present a starting point for the development of novel strategies in cancer treatment and effective diagnosis.

Responses of vascular smooth muscle cells to estrogen are dependent on balance between ERK and p38 MAPK pathway activities.

OBJECTIVE: To investigate the mechanisms underlying the dual effects of estrogen on vascular smooth muscle cells (VSMC). METHODS: MTT assay, ELISA, flow cytometry and Western analysis were used to investigate the effects of 17beta-estradiol (E(2)) on proliferation, apoptosis, cell cycle progression, ERK and p38 activities of subcultured rat VSMC with or without chemical block of MEK or p38 kinases. RESULTS: E(2)-promoted VSMC proliferation was accompanied with an increased phosphorylation of ERK1/2, which could be blocked by MEK inhibitor U0126; the E(2)-induced VSMC apoptosis, which appeared mainly in the G2/M phase, was related with the activation of p38 and could be blocked by p38 inhibitor SB203580. More interestingly, MEK inhibition in E(2)-treated VSMC led to an enhanced p38 phosphorylation and a shift of apoptosis from G2/M phase-predominant to G0/G1 phase-predominant; whereas block of p38 increased the E(2)-induced ERK1/2 phosphorylation and proliferation of the VSMC. This reciprocal phenomenon was related with cross-talk between ERK and p38 pathways which might be mediated by MKP-1 and PP2A. The effects of E(2) on proliferation and apoptosis, and their related pathways could be separately induced by the specific agonists of estrogen receptor (ER) alpha and beta alone and inhibited or eliminated by the ER blocker ICI 182,780. CONCLUSION: The dual effects of estrogen on VSMC involve concurrent activations of ERK and p38 pathways by ER alpha and beta respectively, and the fates of VSMC are determined by the dynamic balance between these two pathways.