Developing Single-Atomic Manganese Nanozymes for Synergistic Mild Photothermal/Multienzymatic Therapy.

Synergistic mild photothermal/nanozyme therapy with outstanding hyperthermia performance and excellent multienzyme properties is highly needed for osteosarcoma treatment. Herein, we have developed efficient single-atom nanozymes (SANs) consisting of Mn sites atomically dispersed on nitrogen-doped carbon nanosheets (denoted as Mn-SANs) for synergistic mild photothermal/multienzymatic therapy against osteosarcoma. Benefiting from their black N-doped carbon nanosheet matrices, Mn-SANs showed an excellent NIR-II-triggered photothermal effect. On the other hand, Mn-SANs with atomically dispersed Mn sites have outstanding multienzyme activities. Mn-SANs can catalyze endogenous H2O2 in osteosarcoma into O2 by catalase (CAT)-like activity, which can effectively ease osteosarcoma hypoxia and trigger the oxidase (OXD)-like catalysis that converts O2 to the cytotoxic superoxide anion radical (•O2-). At the same time, Mn-SANs can also mimic glutathione oxidase (GSHOx) to effectively consume the antioxidant glutathione (GSH) in osteosarcoma and inhibit intracellular glutathione peroxidase 4 (GPX4) expression. Such intratumoral •O2- production, GSH depletion, and GPX4 inactivation mediated by Mn-SANs can create a large accumulation of lipid peroxides (LPO) and •O2-, leading to oxidative stress and disrupting the redox homeostasis in osteosarcoma cells, which can ultimately induce osteosarcoma cell death. More importantly, heat shock proteins (HSPs) can be significantly destroyed via Mn-SAN-mediated plentiful LPO and •O2- generation, thus effectively impairing osteosarcoma cells resistant to mild photothermal therapy. Overall, through the cooperative effect of chemical processes (boosting •O2-, consuming GSH, and enhancing LPO) and biological processes (inactivating GPX4 and hindering HSPs), collaborative mild photothermal/multienzymatic therapy mediated by Mn-SANs is a promising strategy for efficient osteosarcoma treatment.

An intelligent peptide recognizes and traps Mycobacterium tuberculosis to inhibit macrophage phagocytosis.

Tuberculosis is a major public health concern worldwide, and it is a serious threat to human health for a long period. Macrophage phagocytosis of Mycobacterium tuberculosis (M. tuberculosis) is a crucial process for granuloma formation, which shelters the bacteria and gives them an opportunity for re-activation and spread. Herein, we report an intelligent anti-microbial peptide that can recognize and trap the M. tuberculosis, inhibiting the macrophage phagocytosis process. The peptide (Bis-Pyrene-KLVFF-WHSGTPH, in abbreviation as BFH) first self-assembles into nanoparticles, and then forms nanofibers upon recognizing and binding M. tuberculosis. Subsequently, BFH traps M. tuberculosis by the in situ formed nanofibrous networks and the trapped M. tuberculosis are unable to invade host cells (macrophages). The intelligent anti-microbial peptide can significantly inhibit the phagocytosis of M. tuberculosis by macrophages, thereby providing a favorable theoretical basis for inhibiting the formation of tuberculosis granulomas.

MicroRNA-149 rs2292832 C/T Polymorphism Predicts the Prognosis of Hepatocellular Carcinoma Patients With Bone Metastasis.

OBJECTIVE: The prognostic markers of hepatocellular carcinoma (HCC) patients with bone metastasis are of great significance for the design of treatment strategy, the maintenance of life quality of the patients, and the improvement of cancer prognosis. MicroRNA-149 (miR-149) rs2292832 C/T polymorphism in HCC patients has been reported to be associated with the risk of HCC, but whether it can predict the prognosis of HCC patients with bone metastasis remains unclear. The goal of our study was to examine the prognostic impact of miR-149 rs2292832 C/T polymorphism on HCC patients with bone metastasis. METHODS: A total of 67 cases of HCC patients with bone metastasis (BC group) and 73 cases of HCC patients without bone metastasis (NC group) were included in this study. The miR-149 levels in blood leukocytes and tumor tissues were determined by qRT-PCR. Genotyping analysis of miR-149 rs2292832 was performed using polymerase chain reaction (PCR)-restriction fragment length polymorphism assay. RESULTS: The blood leukocyte miR-149 levels were significantly decreased in HCC patients, compared with the healthy controls, and they were significantly decreased in the BC patients, compared with the NC cases. BC patients carrying miR-149 rs2292832 CC+CT phenotype have a better overall survival (OS) rate, whereas no significant correlation was found between miR-149 rs2292832 CC+CT phenotype and the OS rate in NC group. The miR-149 rs2292832 CC+CT phenotype was correlated with certain bone turnover markers and bone metabolism markers but was not correlated with receptor activator of nuclear factor-kappaB ligand (RANKL) expression. Meanwhile, the combination of miR-149 rs2292832 CC+CT phenotype and RANKL expression could improve the prognosis assessment of HCC patients with bone metastasis. CONCLUSION: miR-149 rs2292832 polymorphism might be a novel prognostic biomarker for HCC patients with bone metastasis. A follow-up study with a larger cohort from a multicenter should be performed to test our conclusions.

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.

Potential of bone marrow mesenchymal stem cells in rejuvenation of the aged skin of rats.

The aim of the present study was to evaluate the anti-aging effects of bone marrow-mesenchymal stem cells (BM-MSCs) in a D-galactose-induced skin aging rat model. Male Sprague Dawley rats were randomly divided into four groups (n=10/group) as follows: Normal control group; skin aging model group; MSC-treated group by subcutaneous multi-point injection. The skin aging model was established by a daily subcutaneous injection of 15% D-galactose (1,000 mg/kg) for 8 weeks. Rats in the MSC-treated groups were administered 3×106/ml BM-MSCs/green fluorescent protein (GFP) for 4 weeks, administered once per week. Oxidative/antioxidative parameters were evaluated, and morphological and ultrastructure analyses were performed. Rats in the model group exhibited the typical changes of aging skin. Compared with the control group, rats in the model group had significantly increased malondialdehyde (MDA) content (P<0.01), and decreased serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities (P<0.05). MSC treatment markedly ameliorated aging-induced oxidative stress in the skin. Histologically, rats in the model group exhibited loosely arranged epidermal cell layers and disorganized collagen fibers. BM-MSC treatment significantly improved the histological abnormalities, which was similar to those in the control group. In addition, 7 days after the final cell transplantation, GFP-positive cells were observed by fluorescence microscopy to be distributed in the dermis. Injection of BM-MSCs significantly improved the D-galactose-induced histological abnormalities of the skin, by promoting an antioxidant response and ameliorating oxidative stress in aged skin. Thus, BM-MSCs may be beneficial in the rejuvenation of aged skin.

[Case-control studies of two kinds of method for the treatment of lumbar tuberculosis with psoas abscess].

OBJECTIVE: To compare two kinds of method for treating lumbar tuberculosis with psoas abscess, to provide reference for clinical reasonable select of therapy treatment. METHODS: From January 2010 to January 2013,42 patients with lumbar tuberculosis combined with psoas abscess with obvious surgical indications were enrolled, including 24 males and 18 females with an average age of (38.5 ± 10.2) years old ranging from 21 to 63 years old. All patients were followed up for 18 to 24 months with an average of 20.9 months. Twenty-two patients underwent posterior vertebral body lesions cleared, bone graft fusion and internal fixation and percutaneous puncture catheter drainage for treatment of psoas major abscess as group A, and twenty patients underwent one-stage extraperitoneal approach to remove abscess, posterior vertebral body lesions cleared, bone graft fusion and internal fixation as group B. The operative time, loss of blood, length of hospital stay, clinical cure rate and other clinical results for the two groups were analyzed and compared. RESULTS: The loss of blood was (452.3 ± 137.6) ml in group A and (603.5 ± 99.6) ml in group B, there was significant statistical difference (P < 0.05). The time of operation was (193.6 ± 91.2) min in group A and (230.5 ± 56.6) min in group B, there was significant statistical difference (P < 0.05). The time of operation and the loss of blood in group A were obviously less than which in group B. In group A 20 cases were cured and 2 cases relapsed, 19 cases were cured and 1 case relapsed in group B, there was no significant statistical differences between two groups regarding cure rate with chi-square test (χ² = 0.000, P = 1.000). All patients in two groups obtained good clinical curative effect. There were no significant statistical difference between two groups regarding for length of hospital stay with t-test (P > 0.05). CONCLUSION: Lumbar spinal tuberculosis with psoas abscess is not absolute indications for anterior open operation. Compared with the combined anterior and posterior surgical procedure, the percutaneous puncture catheter drainage combined with posterior debridement, interbody fusion and internal fixation can achieve the same clinical effect but less trauma for the patients.

The classification of recurrent spinal epidural hematoma: a review of the literature and a comparison with the cases.

Symptomatic postoperative spinal epidural hematoma (SEH) and spontaneous spinal epidural hematoma (SSEH) are both rare conditions, and recurrent SEH occurs even less frequently. Therefore, we describe a case of symptomatic postoperative SEH after surgical evacuation of SSEH, which was diagnosed using magnetic resonance imaging (MRI) and managed with negative pressure wound therapy (NPWT). The authors classified the reported recurrent SEHs into two types based on the cause of their previous hematoma, which can be classified as spontaneous or postoperative. The characteristics, diagnosis, managements, and results of recurrent SEHs were analyzed. The authors suggest that the postoperative SEH in the Type II will be treated with NPWT, and the new classification will be helpful for prognosis, diagnosis, and management of the recurrent SEHs.

Refractory Dyslipidemia After Liver Transplant: Case Study With Successive Histologic Investigations.

Hyperlipidemia is not unusual in liver transplant recipients, but refractory severe hyperlipidemia is unusual. We treated a 39-year-old man who had severe dyslipidemia after liver transplant. The levels of blood lipids, liver enzymes, and essential indicators of liver pathology were monitored. The first serum sample was collected from the liver recipient 56 days after transplant surgery because samples could not be obtained sooner after the transplant. The levels of liver enzymes and blood lipids were improved with symptomatic treatment but had recurrent fluctuations. Tacrolimus and cyclosporine, even at low doses, may have been the dominant factor affecting the blood lipid levels in the recipient.

Clinical and molecular characteristics of Chinese patients with X-linked lymphoproliferative syndrome type 1.

BACKGROUND: X-linked lymphoproliferative syndrome type 1 (XLP1) is a rare inherited, life-threatening immunodeficiency disorder caused by mutations in SH2D1A gene. It affect approximately two to three males per million. Fewer than 10 cases with definite gene mutations have been reported in Chinese mainland and no rapid diagnosis method has been established. PROCEDURE: We determined the clinical and molecular characteristics of five patients with XLP1. The SH2D1A gene were amplified by PCR and sequenced, the SAP expression was analyzed by flow cytometry. RESULTS: Two patients had novel SH2D1A mutations and three had mutations that have been previously reported. Three patients presented with fulminant infectious mononucleosis or hemophagocytic lymphohistiocytosis and one presented with lymphoma. Null or decreased SAP expression on PBMCs was noted. The remaining patient presented with unique, recurrent, nonfulminant infectious mononucleosis and bimodal intracellular SAP protein expression. CONCLUSIONS: The overall molecular characteristics and clinical phenotypes of Chinese patients with XLP1 matched previous reports. The unique bimodal intracellular SAP protein expression indicated the presence of some residual SAP-positive T cells that are able to respond to persistent Epstein-Barr virus infection and could explain the relatively mild clinical phenotype of this patient.

TCAB1: a potential target for diagnosis and therapy of head and neck carcinomas.

BACKGROUND: WRAP53, including α, ß and γ isoforms, plays an important role not only in the stability of p53 mRNA, but also in the assembly and trafficking of the telomerase holoenzyme. It has been considered an oncogene and is thought to promote the survival of cancer cells. The aim of this study was to detect the role of TCAB1 (except WRAP53α) in the occurrence and development of head and neck carcinomas. METHODS: Immunohistochemistry was used to detect the TCAB1 expression in clinical specimen sections and performed western blotting to check the TCAB1 expression levels in cell lines. TCAB1 was depleted using shRNA lentivirus and the knockdown efficiency was assessed using q-PCR and Western blotting. We performed CCK-8 assays and flow cytometry to check the cell proliferation potential and used the trans-well assay to test the invasion ability in vitro. Xenografts were used to detect the tumor formation potential in vivo. Moreover, we performed cDNA microarray to investigate the candidate factors involved in this process. RESULTS: We observed a notable overexpression of TCAB1 in head and neck carcinoma clinical specimens as well as in carcinoma cell lines. Knockdown of TCAB1 decreased the cellular proliferation potential and invasion ability in vitro. cDNA microarray analysis suggested the possible involvement of several pathways and factors associated with tumorigenesis and carcinoma development in the TCAB1-mediated regulation of cancers. Furthermore, the xenograft assay confirmed that the depletion of TCAB1 would inhibit tumor formation in nude mice. The immunohistochemistry results of the mice tumor tissue sections revealed that the cells in shTCAB1 xenografts showed decreased proliferation potential and increased apoptotic trend, meanwhile, the angiogenesis was inhibited in the smaller tumors form shTCAB1 cells. CONCLUSIONS: Our study demonstrated that depletion of TCAB1 decreased cellular proliferation and invasion potential both in vitro and in vivo. The data indicated that TCAB1 might facilitate the occurrence and development of head and neck carcinomas. In future, TCAB1 might be useful as a prognostic biomarker or a potential target for the diagnosis and therapy of head and neck carcinomas.

[Application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease].

OBJECTIVE: To explore the value of application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease. METHODS: Clinical datas of 13 patients with multi-segment lumbar degenerative disease (8 males and 5 females,ranging in age from 51 to 72 year with an average of 65.0) were retrospectively analyzed between April 2008 and May 2009. The involved area included L3-S1 in 7 cases, L2-S1 in 3 cases, L3-L5 in 1 cases, L4-S1 in 2 cases. All patients underwent decompression, dynamic stabilization with Bioflex system, according to the severity of degenerative disc with/without interbody fusion. The clinical effects were evaluated by VAS, ODI. ROM and fusion segments were also observed. RESULTS: The mean follow up period was 19.5 months (from 12 to 26 months). The mean operative time was 183.4 min (from 90 to 240 min) and the mean volume of blood loss was 610.2 ml (from 400 to 1 220 ml). The mean VAS score was 7.8 +/- 1.3 preoperatively, 2.3 +/- 0.9 postoperatively and 2.1 +/- 0.8 at the last follow up. The average ODI was (60.50 +/- 4.40)% preoperatively, (17.80 +/- 2.10)% postoperatively and (16.20 + 2.40)% at the last follow up. The VAS and ODI significant improved in postoperatively (P < 0.05), and there was no statistical difference between postoperative and last follow up (P > 0.05). ROM of whole lumbar and non-fused segment showed obviously decreased and adjacent segment showed insignificant increased. The fusion rate of interbody fusion level was 95.0% (19/20). CONCLUSION: The preliminary clinical results show the Bioflex system combined with intebody fusion is a safe and effective technique in treating multi-segment lumbar degenerative disease.

[Clinical study of short-course chemotherapy combined with radical operation in retreating spinal tuberculosis].

OBJECTIVE: To investigate clinical outcome of short-course chemotherapy in retreating spinal tuberculosis after radical operation. METHODS: Forty-six retreating patients with spinal tuberculosis were included in this series, 29 males, 17 females with the age from 27 to 61 years (average of 43.7 years). All patients were treated with radical operation and short-course anti-tuberculous chemotherapy from March 2005 to March 2008. The tuberculous focus located thoracic spine in 17 cases, thoracic-lumbar in 13 and lumbosacral vertebrae in 16 cases. Of them, 5 cases had sinuses of tuberculosis and 7 cases had incomplete palsy in lower limbs (Frankel C-D). CT or MRI showed obvious sequestra, cold abscess within spinal focus. Surgical procedures including debridement, auto-bone grafting, and one-stage internal fixation, was performed at the 4 to 6 weeks after chemotherapy. Chemotherapy regimes were 3HRZ/6-9HRE in majority of patients. Clinical effect and focus healing were evaluated at follow-up period. RESULTS: Tuberculous symptoms and local pain of vertebral volume were obvious in all patients before chemotherapy,with average ESR 65.3 mm/h and average CRP 37.4 mg/L. After 4-6 weeks chemotherapy, tuberculosis symptoms and vertebral pain improved in all patients, and the average ESR decreased to 38.3 mm/1h, the average CRP decreased to 17.2 mg/L. Two to three months after operation, tuberculous symptoms and local pain relived in all patients,ESR and CRP became normal in 37 cases. Six to twelve months after operation, bonegraft complex in each patient became stable and there were no instrument loosening or deformity correction loss. Six patients with incomplete palsy recovered and 1 case improved from Frankel C to D grade. Focus healing was achieved in 44 cases (95.7%) after short-course chemotherapy (3HRZ/6-9HRE), and there were no resurgence in 2 to 4 years follow-up period. Drug fast 2 cases for RFP+INH cured at the 15 months after chemotherapy. CONCLUSIONS: Removed tubercular focus for the treatment of retreating spinal tuberculosis can improve clinical effect and shorten chemotherapy course.

[Variety of ESR and C-reactive protein levels during perioperative period in spinal tuberculosis].

OBJECTIVE: To investigate the clinical significance of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in the treatment of spinal tuberculosis. METHODS: Sixty-seven patients (41 males and 26 females, ranging in age from 23 to 61 years) with active spinal tuberculosis in our hospital (from Mar. 2004 to Mar. 2007) were included in this study. The tuberculosis focus were located either in cervical spine, thoracic spine or in lumbar spine. After 4 to 6 weeks anti-tuberculosis chemotherapy, all the patients underwent one-stage operation (focus debridment) and auto-bone graft combined with internal fixation. Blood test for ESR and CRP were carried out at different times before and after operation. RESULTS: The average ESR was (79.4 +/- 35.6) mm/h, and the average CRP was (44.3 +/- 17.5) mg/L before chemotherapy, indicating active tuberculosis focus. After 4 to 6 weeks chemotherapy, the average ESR was (45.3 +/- 21.0) mm/h,and the average CRP was (26.7 +/- 11.8) mg/L, the differences were statistically (P < 0.05), and the clinical symptoms of spinal tuberculosis relieved in all patients. Four weeks after operation, the average ESR dropped to (42.8 +/- 16.5)mm/h, the average CRP dropped to (23.8 +/- 10.0) mg/L statistically (P < 0.05). Eight weeks after operation, the average value of ESR and CRP were at normal level in 47 cases, indicating inactive tuberculosis focus. Focus healing was achieved in 65 patients after short-term chemotherapy. CONCLUSION: The level of ESR and CRP are high in active spinal tuberculosis and low when focus controlled. ESR and CRP are reliable parameters in evaluation the treatment and prognosis of spinal tuberculosis.