Negative pressure promotes macrophage M1 polarization after Mycobacterium tuberculosis infection via the lncRNA XIST/microRNA-125b-5p/A20/NF-κB axis.

Experiments have demonstrated the regulation of long noncoding RNA (lncRNA) in tuberculosis (TB), and negative pressure treatment has been associated with the alleviation of TB. Here, we investigated the interaction of negative pressure and the lncRNA X-inactive specific transcript (XIST) in modulating Mycobacterium tuberculosis (MTB) infection. Initially, we established an in vitro cell model of MTB infection and an in vivo mouse model of MTB infection, followed by treatment with negative pressure. Then, we examined the expression of XIST, followed by analysis of the downstream miRNA of XIST. XIST was overexpressed or underexpressed through cell transfection to examine its effects on macrophage polarization via the miR-125b-5p/A2 axis. The MTB models were characterized by upregulated XIST and downregulated miR-125b-5p. XIST bound to miR-125b-5p, leading to its downregulation, and thus causing higher MTB survival in an ESAT-6-dependent manner. Additionally, negative pressure treatment decreased MTB-driven XIST expression through downregulation of A20 (an NF-κB repressor) via miR-125b-5 expression, promoting the M1 polarization program in macrophages through activation of the NF-κB pathway. In summary, negative pressure treatment after MTB infection can promote the polarization of macrophages to the proinflammatory M1 phenotype by regulating the XIST/miR-125b-5p/A20/NF-κB axis.

Proinflammatory Effects of Ubiquitin-Specific Protease 5 (USP5) in Rheumatoid Arthritis Fibroblast-Like Synoviocytes.

Rheumatoid arthritis (RA) is a worldwide chronic autoimmune inflammatory disease which is affecting approximately 1% of the total population. It is characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS) and increased production of proinflammatory cytokines. In the current study, we were aiming to investigate the role of ubiquitin-specific protease 5 (USP5) in the inflammatory process in RA-FLS. Expression of USP5 was found upregulated in RA-FLS compared with that in osteoarthritis- (OA-) FLS, and IL-1ß stimulation increased USP5 expression in a time-dependent manner. Furthermore, we found that USP5 overexpression significantly aggravated proinflammatory cytokine production and related nuclear factor κB (NF-κB) signaling activation. Consistently, silencing of USP5 decreased the release of cytokines and inhibited the activation of NF-κB. In addition, USP5 was found to interact with tumor necrosis factor receptor-associated factor 6 (TRAF6) and remove its K48-linked polyubiquitination chains therefore stabilizing TRAF6. Our data showed that a USP5-positive cell regulates inflammatory processes in RA-FLS and suggested USP5 as a potential target for RA treatment.

The PI3K/AKT cell signaling pathway is involved in regulation of osteoporosis.

BACKGROUND: Osteoporosis is a systemic skeletal disease with the high incidence, serious complications, financial burden, and heavily decrease in living quality. METHODS: Proliferation of osteoblast was tested by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method, alkaline phosphatase (ALP) activity of osteoblasts was tested by ALP REAGENT, Calcium level was determined by a colorimetric assay, mRNA expression of phosphoinositide-3 kinase (PI3K), 3-phosphoinositide-dependent protein kinase 1 (PDK1), Akt, Caspase-3, Caspase-7, Caspase-9, osteocalcin (OCN), Osterix and Runx2 of osteoblasts was tested by RNA preparation and quantitative reverse transcription polymerase chain reaction (RT-PCR), and protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt was measured by Western Blot analysis. RESULTS: In osteoporosis model rats, it found that mRNA expression of PI3K, PDK1 and Akt showed no changes while protein expression of phospho-PI3K, phospho-PDK1 and phospho-Akt in bone tissue was decreased dramatically. To further characterize the molecular mechanisms that regulate osteoporosis, we examined the contribution of the PI3K/Akt cell signaling pathway in cultured osteoblasts. It suggested that, the blockade of PI3K activation by LY294002, a specific inhibitor of the PI3K/Akt signaling pathway in osteoblasts, heavily inhibited cell proliferation, ALP activity, calcium accumulation, and mRNA expression of OCN, Osterix and Runx2. However, mRNA expression of Caspase-3 and Caspase-9 was promoted accordingly. CONCLUSION: The in vivo and in vitro studies indicated that the PI3K/Akt cell signaling pathway is involved in the inhibition of osteoporosis through promoting osteoblast proliferation, differentiation and bone formation.

[Diagnosis and treatment of lumbar disc herniation by discography and percutaneous transforaminal endoscopic surgery].

OBJECTIVE: To conduct a retrospective analysis of diagnosis and treatment of lumbar disc herniation by discography and percutaneous transforaminal endoscopic surgery. METHODS: From December 2009 to June 2010, 119 patients with lumbar disc herniation underwent discography and transforaminal endoscopic surgery under local anesthesia. There were 75 males and 44 females with a mean age of 44.8 years (range: 15 - 55). The mean disease course was 9 months (range: 3 - 72). The major symptoms were back pain and/or unilateral sciatica. The mean follow-up period was 26 months. All underwent lumbar radiography, computed tomography (CT) and magnetic resonance imaging (MRI) revealing 112 single level and 7 two-level disc herniations. There were 82 lateral and 37 para-medial disc herniations. Eight-nine patients had protruded discs while 30 had prolapsed and sequestered discs. There were no obvious lumbar stenosis, spondylolisthesis, fracture, infection or tumor cases. The preoperative and postoperative visual analogue scale (VAS) were used to evaluate the sciatica and/or back pain. The outcomes were evaluated by Oswestry disability index (ODI) and the Macnab score. Precise orientation and operation was performed under the guidance of pre-operative imaging, intra-operative fluoroscopy or CT and endoscopic exploration. RESULTS: Among them, 117 cases had the surgery performed successfully. The mean operative duration was 85 min (range: 35 - 85) and the mean blood loss 13 ml (range: 1 - 50). One patient had L5 nerve root injury complicated with paraesthesia and weakness of the affected lower extremity and was relieved gradually after conservative treatment for over 3 months. Another one complicated with postoperative intradiscal infection was referred to another institution and lost follow-up thereafter. Five cases had no improvement at 6 months after the first surgery and were re-operated endoscopically. No one had a conversion into open surgery. They were followed up for a mean period of 26.1 months (range: 25 - 27). Five patients lost follow-up. VAS improved statistically significantly from preoperative 6.8 to postoperative 1.8 (P < 0.05). ODI decreased from preoperative 70.06 to 19.09 at the last follow-up. The Macnab results were excellent (n = 82, 68.9%), good (n = 20, 16.7%), fair (n = 8, 6.7%) and bad (n = 9, 7.7%) (including all patients lost to follow-up). And the excellent-to-good rate was 85.6%. CONCLUSION: With fewer complications and a low recurrence rate, percutaneous transforaminal endoscopic surgery is safe and efficacious in the treatment of lumbar disc herniation.

[Changes in tumor necrosis factor-alpha mRNA expression in peripheral polymorphonuclear leukocyte and tissues during posttraumatic sepsis].

OBJECTIVE: To evaluate the changes in tumor necrosis factor-alpha (TNF-alpha) mRNA expression in peripheral polymorphonuclear leukocyte (PMNs) and tissues after cecal ligation puncture (CLP) in rats. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect TNF-alpha mRNA expression in peripheral PMNs and tissues. RESULTS: The TNF-alpha mRNA expression in peripheral PMNs rose gradually after CLP, and it began to decrease after reaching the peak at 48 hours, but it was still higher than normal. The elevation of TNF-alpha mRNA expression was first limited in the regional tissues (P<0.01 at 12 hours after CLP and peaked at 24 hours in the intestine), then it entered the blood circulation later to affect the sensitive organs--lungs and livers (P<0.01 at 24 hours after CLP and peaked at 48 hours in the lung; P<0.05 at 24 hours after CLP and peaked at 48 hours in the liver). CONCLUSION: The rise of TNF-alpha mRNA expression plays an important role in pathogenesis of sepsis.

[Apoptosis of neutrophil in peripheral blood in experimental sepsis].

OBJECTIVE: To evaluate the changes in apoptosis of neutrophil in peripheral blood in sepsis in rats. METHODS: The rat sepsis model was reproduced by cecum ligation and puncture (CLP). One hundred and forty-four rats were randomly divided into normal control group, sham operation group and 2, 6, 12, 24, 48, 72 hours after CLP groups. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to identify neutrophil apoptosis. RESULTS: In early period after CLP, neutrophil apoptosis in peripheral blood was limited with a positive rate of less than 5.00%. The positive rate rose to (48.33+/-12.53)% at 48 hours, and it began to lower, approaching the normal level at 72 hours after CLP. CONCLUSION: Death is the main pathway of loss of neutrophils which are produced in the acute phase of sepsis, and apoptosis is the main pathway of loss of neutrophil in the later phase of sepsis.