Advantages of unilateral percutaneous kyphoplasty for osteoporotic vertebral compression fractures-a systematic review and meta-analysis.

Data from English randomized controlled trials comparing unilateral versus bilateral PKP for the treatment of OVCFs were retrieved and analyzed, and the results showed that unilateral PKP is a better choice for the treatment of patients with OVCFs, which will provide a reliable clinical rationale for the treatment of OVCFs. PURPOSE: To investigate the advantages of unilateral percutaneous kyphoplasty (PKP) for the treatment of osteoporotic vertebral compression fractures(OVCFs). METHODS: The systematic evaluation program met all program requirements (CRD 42023422383) by successfully passing the PROSPERO International Prospective Systematic Evaluation Registry. Researchers searched the references of English-language randomized controlled trials comparing unilateral and bilateral PKP for the treatment of osteoporotic vertebral compression fractures published between 2010 and 2023 and manually searched for known primary and review articles. The study statistically analyzed data from all the included literature, which primarily included time to surgery, visual pain score(VAS) and Oswestry disability index(ODI) at postoperative follow-up time points, polymethylmethacrylate (PMMA, bone cement) injection dose, cement leakage, radiation dose, and improvement in kyphotic angle. RESULTS: This meta-analysis searched 416 articles published from 2010 to 2023 based on keywords, and 18 articles were finally included in this study. The results of the forest plot showed that unilateral PKP operative time, amount of bone cement used, and radiation dose to the patient were significantly reduced (p < 0.01, p < 0.01, and p < 0.01, respectively), and unilateral and bilateral PKP had comparable cement leakage (p = 0.49, 95% CI = 0.58-1.30), and there was no significant difference in the kyphotic angle between unilateral and bilateral PKP (p = 0.42, 95% CI = - 2.29-0.96). During follow-up, there was no significant difference in pain relief between unilateral and bilateral PKP (p = 0.70, 95% CI = - 0.09-0.06), nor was there a significant difference in ODI (p = 0.27, 95% CI = - 0.35-1.24). CONCLUSIONS: There is no difference in clinical efficacy between unilateral PKP and bilateral PKP, but unilateral PKP has a shorter operative time, a lower incidence of cement leakage, a lower amount of cement, and a lower radiation dose to the patient and operator. Unilateral PKP is a better option for patients with OVCFs.

Inhibition of miR-96-5p alleviates intervertebral disc degeneration by regulating the peroxisome proliferator-activated receptor γ/nuclear factor-kappaB pathway.

BACKGROUND: Intervertebral disc degeneration (IDD) is the main pathogenesis of low back pain. MicroRNAs (miRNAs) have been found to exert regulatory function in IDD. This study aimed to investigate the effect and potential mechanism of miR-96-5p in IDD. METHODS: In vitro cell model of IDD was established by treating human nucleus pulposus cells (HNPCs) with interleukin-1ß (IL-1ß). The level of peroxisome proliferator-activated receptor γ (PPARγ) was examined in the IDD cell model by Western blot and quantification real-time reverse transcription-polymerase chain reaction (qRT-PCR). The expression level of miR-96-5p was detected by RT-qPCR. Effects of PPARγ or/and PPARγ agonist on inflammatory factors, extracellular matrix (ECM), apoptosis, and nuclear factor-kappaB (NF-κB) nuclear translocation were examined through enzyme-linked immunosorbent assay (ELISA), Western blot, flow cytometry assay, and immunofluorescence staining. The Starbase database and dual luciferase reporter assay were used to predict and validate the targeting relationship between miR-96-5p and PPARγ, and rescue assay was performed to gain insight into the role of miR-96-5p on IDD through PPARγ/NF-κB signaling. RESULTS: PPARγ expression reduced with concentration and time under IL-1ß stimulation, while miR-96-5p expression showed the reverse trend (P < 0.05). Upregulation or/and activation of PPARγ inhibited IL-1ß-induced the increase in inflammatory factor levels, apoptosis, degradation of the ECM, and the nuclear translocation of NF-κB (P < 0.05). MiR-96-5p was highly expressed but PPARγ was lowly expressed in IDD, while knockdown of PPARγ partially reversed remission of IDD induced by miR-96-5p downregulation (P < 0.05). MiR-96-5p promoted NF-κB entry into the nucleus but PPARγ inhibited this process. CONCLUSION: Inhibition of miR-96-5p suppressed IDD progression by regulating the PPARγ/NF-κB pathway. MiR-96-5p may be a promising target for IDD treatment clinically.

NEP1-40-overexpressing Neural Stem Cells Enhance Axon Regeneration by Inhibiting Nogo-A/NgR1 Signaling Pathway.

BACKGROUND: Nogo-66 antagonistic peptide (NEP1-40) offers the potential to improve spinal cord injury (SCI). OBJECTIVE: To explore the effect of NEP1-40 overexpression on neural stem cells (NSCs) regulating the axon regeneration of injured neurons. METHODS: We isolated NSCs from brain tissues of pregnant rat fetuses and used Nestin immunofluorescence to identify them. The NEP1-40 overexpressing NSCs were constructed by transfection with the NEP1-40-overexpressing vector. The expression of NSCs differentiation associated markers, including Tuj-1, GFAP, Oligo2, and MBP, were detected by RT-PCR, western blotting, and immunofluorescence. NeuN immunofluorescence staining was used to measure the number of neurons. And western blotting was used to detect the phosphorylation levels of LIMK1/2, cofilin, and MLC-2 and the protein levels of GAP-43, MAP-2, and APP. RESULTS: The NEP1-40 overexpression promoted the expression level of Tuj-1, Oligo2, and MBP, and increased the number of Tuj-1, Oligo2, and MBP positive cells. NEP1-40-overexpressing NSCs (NEP-NSCs) improved NeuN positive cells of co-culture with injured neurons. And NEP-NSCs also increased the protein levels of axon regeneration indicators (GAP-43, MAP-2) and decreased APP protein level. In addition, the phosphorylation level of LIMK1/2, cofilin, and MLC-2 were markedly decreased in NEP-NSCs. CONCLUSION: NEP1-40 overexpression enhanced the ability of NSCs differentiation into neurons and promoted axon regeneration by inhibiting the Nogo-A/NgR1 signaling pathway. This study provides an alternative gene modified transplantation NSCs for the SCI treatment.

Obesity is a risk factor for epidural lipomatosis: a meta-analysis.

PURPOSE: The purpose of this meta-analysis was to summarize the evidence of associations between obesity factors and spinal epidural lipomatosis (SEL) and to evaluate the strength and validity of these associations. METHODS: Electronic databases such as Wiley Online Library, PubMed, Embase, Cochrane Library were searched and manual retrieval of references, the time limit was from the establishment of the database to May 2020. Methodological quality evaluations of the included studies were assessed using the bias risk assessment tool recommended by the Cochrane Guidelines. The RevMan 5.3 software was used for meta-analysis. RESULTS: Finally, seven studies were included for meta-analysis, all of which were observational studies with mixed bias risk. These studies involved 807 patients, with an average age of 64 to 73.6 years, and 59.4 percent of the participants were male. The sample sizes for the included studies ranged from 28 to 288. The results of meta-analysis showed that high body mass index (BMI) was one of the factors affecting SEL (P < 0.01, MD 1.37, 95% CI [0.81, 1.92]). All reviews had a high risk of bias, and the most common source of bias was that there was no strict unified case diagnosis standard between researches, and some studies (four items) did not clearly describe the confounders that they controlled. CONCLUSIONS: We suggest that physicians should consider high BMI as a factor leading to SEL, and to control body weight actively should be considered as the preferred treatment strategy before surgical intervention is conducted.

Does surgical treatment increase the progression of spinal cord injury in patients with ossification of posterior longitudinal ligament of cervical spine? A systematic review and meta-analysis.

PURPOSE: The purpose of this study is to explore if the surgical treatment will accelerate the progression of spinal cord injury (SCI) in patients with cervical Ossification of the posterior longitudinal ligament (OPLL) and if surgery will have better curative effect than conservative treatment. METHODS: An extensive search of literature was implemented in PubMed, EMBASE, and other online databases. The quality of the included articles was evaluated according to the Newcastle-Ottawa Quality Assessment Scale, as recommended by the Cochrane manual, and meta-analysis was performed using the review manage5.3 software. RESULTS: No obvious statistical difference was observed in the rate of SCI progression (P > 0.05, OR 1.15 [0.66, 2.00]), cervical range of motion, (P > 0.05, weighted mean difference (WMD) 4.52 [-5.75, 14.79]), and Japanese Orthopedic Association scores before surgery (P > 0.05, WMD -2.78 [-7.87, 2.32]) between the surgical group and conservative treatment group. However the surgical group illustrated obviously higher neurofunctional recovery rate (P < 0.05, OR 6.07 [1.55, 23.78]) and postoperative JOA score of the surgery group (P < 0.05, WMD -0.77 [-1.21, -0.33]) than conservative group. CONCLUSIONS: Based on this meta-analysis, there is not enough evidence to indicate that surgery will accelerate the progress of SCI with OPLL. However, the superiority of surgical efficacy can be observed over conservative treatment in terms of relieving neurological symptoms.

Iron and silicon oxide doped/PAN-based carbon nanofibers as free-standing anode material for Li-ion batteries.

The advanced rechargeable batteries have been upgraded by the production of free-standing electrodes through electrospinning technology. This study aimed to introduce a novel hybrid composition to design a free-standing carbon nanofiber (CNF) based anode by the addition of iron acetylacetonate Fe(AcAc)3 as active material and TEOS(tetraethoxysilane)/APTES(aminopropyl triethoxysilane) as an additive for Li-ion batteries. Polyacrylonitrile(PAN) was used as the polymer matrix in the spin dope, and the inclusion of Fe(AcAc)3 and TEOS/APTES resulted in compositional change, producing iron oxide and silica nanoparticles throughout the matrix. Different oxidation states and the presence of embedded iron oxide nanoparticles in CNF were identified by XPS and EDX elemental mapping analysis. PAN-TEOS-APTES-Fe(AcAc)3 based anode material was capable of enhancing the reversible specific capacity as much as 732 mAhg-1 at 500 mAg-1 lasting for 300 cycles, along with the rate capability as high as 815 mAhg-1 at 200 mAg-1. Furthermore, EIS analysis and EX-situ FESEM showed decreased impedance after cycling with the stable morphology of PAN-TEOS-APTES-Fe(AcAc)3 CNF-film. Conclusively, non-woven, binder-free, current collector free, free-standing CNF anode film doped with iron-oxide and silica nanoparticles was indeed a novel approach for Li-ion batteries and can be considered for other batteries particularly for Li-S batteries.

K-Ras-PI3K regulates H3K56ac through PCAF to elevate the occurrence and growth of liver cancer.

H3 modification is related to a wide range of tumors, including liver cancer. The Ras passageway is actuated in human diseases. Thus, we investigated the roles of Ras in liver cancer cells via acetylation of H3K56. Ras-carrying G12V and Y40C site mutation was transfected into liver cancer cell lines SNU-475 and SK-Hep-1. Acetylation of H3K56 and phosphatidylinositol 3-kinase (PI3K), P300/CBP-associated factor (PCAF) and Mouse double minute 2 homolog (MDM2) was tested via western blot. Cell activity, colonies, and migration were tested via Cell Counting Kit-8, soft-agar colony formation, and Transwell experiment, respectively. Sirtuin 6 (SIRT6) and PCAF were tested via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Chromatin immunoprecipitation was employed to test the relationship between Ras and downstream elements. Flow cytometry was employed to test cell cycle series. We found that RasG12V/Y40C transfection reduced the acetylation of H3K56 and activated phosphorylation of protein kinase B. H3K56Q (H3K56ac overexpression) suppressed cell activity, colonies, and migration. H3K56ac changed Ras downstream factors expression. RasG12V/Y40C bound to Ras-PI3K downstream elements' promoters. SIRT6 silencing raised H3K56ac and suppressed cell activity, migration and S phage cell percentage. SIRT6 silence transformed expression of downstream elements. PCAF and H3K56ac demonstrated the close current while MDM2 was conversed. In summary, the Ras-PI3K passageway promoted cell growth and metastasis via decreasing H3K56ac, in which MDM2-mediated PCAF was involved.

Sinomenine exerts antitumour effect in gastric cancer cells via enhancement of miR-204 expression.

Gastric carcinoma (GC) is a pernicious neoplasm with high morbidity and mortality. Sinomenine (SIN) has long been exploited to heal rheumatoid arthritis. Recently, SIN has been discovered to exert the antitumour functions in diverse cancers. However, the impacts of SIN on GC remain indistinct. We attempted to expose the antitumour effect of SIN on GC. MKN45 and SGC-7901 cells were administered with SIN for 24 hours, cell viability, proliferation, apoptosis, migration, invasion and the associated proteins in the above processes were examined via exploiting CCK-8, BrdU, flow cytometry, Transwell and Western blot. MiR-204 expression in GC tumour tissues, different GC cell lines and SIN-stimulated GC cells was investigated by executing RT-qPCR. The above cell biological processes were reassessed after transfection with miR-204 inhibitor. The latent mechanisms were probed by examining AMPK and Wnt/ß-catenin pathways. We found that SIN memorably repressed cell proliferation, evoked apoptosis and affected CyclinD1, Bcl-2, Bax and cleaved-caspase-3 expression in MKN45 and SGC-7901 cells. Cell migration, invasion and expression of MMP-9 and Vimentin were all restrained by SIN stimulation. The increase of miR-204 was discovered in GC tissues and SIN-treated MKN45 and SGC-7901 cells. But suppression of miR-204 was observed in AGS, MKN28, MKN45 and SGC-7901 cells. Suppression of miR-204 overturned the inhibitory functions of SIN in MKN45 and SGC-7901 cells. Besides, SIN prohibited AMPK and Wnt/ß-catenin pathways via enhancement of miR-204. In conclusion, these findings suggested that SIN exerted the antitumour activity in GC cells by hindering AMPK and Wnt/ß-catenin pathways via enhancement of miR-204.

[Application of small incision approach in anterior surgery of thoracic and lumbar spinal tuberculosis].

OBJECTIVE: To investigate the value of small incision approach in the anterior surgery of thoracic and lumbar spinal tuberculosis. METHODS: A clinical data of 65 patients with thoracic or lumbar spinal tuberculosis treated with posterior-anterior surgery between January 2015 and January 2018 was retrospectively analyzed. The patients were divided into small incision group (group A, 29 patients) and traditional incision group (group B, 36 patients) according to the length of anterior incision. There was no significant difference in general data such as gender, age, disease duration, segment of lesion, American Spinal Cord Injury Association (ASIA) grading, preoperative pain visual analogue scale (VAS) score, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and Cobb angle of spinal kyphosis between 2 groups ( P>0.05). The length of anterior incision, operation time, intraoperative blood loss, postoperative complications, postoperative hospitalization time, ESR, and CRP were recorded and compared. The VAS score was used to evaluate the pain after operation. The Cobb angles in patients with spinal kyphosis were measured and the loss of angle and correction rate of angle were calculated. The result of bone graft fusion was assessed according to the Bridwell standard. RESULTS: The length of anterior incision, operation time, intraoperative blood loss, and hospitalization time of group A were all significantly less than those of group B ( P<0.05). All patients in both groups were followed up 12-29 months (mean, 20 months). There were 4 cases (13.8%) and 14 cases (38.9%) of postoperative complications in groups A and B respectively, showing significant difference ( χ 2=5.050, P=0.025). The ESR and CRP in 2 groups all returned to normal at 6 months after operation, and there was no significant difference in ESR and CRP between 2 groups at 3 months, 6 months, and last follow-up ( P>0.05). At last follow-up, the neurological function of patient with neurological symptoms was significantly better than that before operation, and there was no significant difference between 2 groups ( Z=0.167, P=0.868). The VAS scores of 2 groups at each time point after operation were significantly lower than those before operation ( P<0.05); the VAS score in group A was significantly lower than that in group B ( t=-2.317, P=0.024) at 1 day after operation, but there was no significant difference between 2 groups ( t=-0.862, P=0.392) at last follow-up. Among the patients with kyphosis, the Cobb angle was significantly decreased at 1 day after operation and last follow-up when compared with preoperative angle ( P<0.05); but there was no significant difference between 1 day after operation and last follow-up ( P>0.05). There was no significant difference in Cobb angle, loss of angle, and correction rate between 2 groups after operation ( P>0.05). The bone graft healed well at last follow-up in 2 groups. There was no significant difference in bone graft fusion rate between 2 groups at 6 months after operation, 1 year after operation, and last follow-up ( P>0.05). At last follow-up, all patients cured, and no recurrence occurred. CONCLUSION: In the anterior surgery of thoracic and lumbar tuberculosis, the application of small incision approach can achieve the similar effectiveness as traditional incision surgery with the advantages of minimally invasive, less complications, and quick recovery.

Effect of T helper cell 1/T helper cell 2 balance and nuclear factor-κB on white matter injury in premature neonates.

Incidence of white matter injury (WMI), which is featured as softening of white matter tissues, has recently increased. Previous studies have demonstrated a close correlation between T helper cell 1 and T helper cell 2 (Th1/Th2) imbalance and nuclear factor­κB (NF­κB) with brain disease. Their role in premature WMI, however, remains to be illustrated. Serum samples were collected from 60 premature WMI neonates, plus another control group of 60 premature babies without WMI. Patients were further divided into mild, moderate and severe WMI groups. Reverse transcription quantitative polymerase chain reaction was used to test mRNA expression levels of Th1/Th2 cytokines, including interleukin 2 (IL)­2, tumor necrosis factor­α (TNF­α), IL­4, IL­10 and nuclear factor (NF)­κB, whilst their serum levels were measured by ELISA. Their correlation with disease occurrence and progression were further analysed, to illustrate the effect of Th1/Th2 balance and NF­κB on pathology of premature WMI. Serum levels of IL­4 and IL­10 were significantly decreased in premature WMI babies, whilst IL­2, TNF­α and NF­κB were upregulated (P<0.05 vs. control group). With aggravated disease, IL­4 and IL­10 expression was further decreased while IL­2, TNF­α and NF­κB were increased (P<0.05 vs. mild WMI group). Th1 cytokines IL­2 and TNF­α and NF­κB were negatively correlated with Th2 cytokines IL­4 and IL­10. Disease severity was positively correlated with IL­2, TNF­α and NF­κB expression, and was negatively correlated with IL­4 and IL­10 (P<0.05). Th1/Th2 imbalance and NF­κB upregulation were observed in WMI pathogenesis, with elevated secretion of Th1 cytokines and decreased Th2 cytokines, suggesting that Th1/Th2 imbalance and NF­κB upregulation may be a potential indicator for the early diagnosis and treatment of WMI pathogenesis and progression.

Protective effect of miconazole on rat myelin sheaths following premature infant cerebral white matter injury.

The aim of the present study was to investigate the protective effects of miconazole on myelin sheaths following cerebral white matter damage (WMD) in premature infant rats. Sprague Dawley rats (3-days-old) were randomly divided into four groups (n=30 each) as follows: Sham surgery group, WMD model group, 10 mg/kg/day treatment group and 40 mg/kg/day treatment group. A cerebral white matter lesion model was created by ligating the right common carotid artery for 80 min. Treatment groups were administered with 10 or 40 mg/kg miconazole at 4-8 days following birth (early treatment group) or 5-11 days following birth (late treatment group). Rats in the model group received the same concentration of dimethylsulfoxide. Myelin basic protein (MBP) immunohistochemical staining and western blotting were used to detect the expression of cerebral white matter-specific MBP, and changes in myelin structure were observed using transmission electron microscopy. No swelling or necrosis was observed in the corpus callosum of the sham group rats, whereas rats in the model group demonstrated edema, loose structure, fiber disorder, inflammatory gliocytes and selective white matter lesions. Following treatment with miconazole, MBP expression in the corpus callosum was significantly higher compared with the model group. Furthermore, in the model group, myelin sheaths in the corpus callosum were loose with small vacuoles, there was a marked decrease in thickness and structural damage was observed. Conversely, a marked improvement in myelination was observed in the treatment group. The results of the present study suggest that miconazole is able to promote formation of the myelin sheath to ameliorate premature cerebral white matter lesions caused by ischemia or hypoxia in rats.

Sudden onset of static equilibrium dysfunction in patients receiving a cochlear implant.

BACKGROUND: We investigated the sudden onset of static equilibrium dysfunction caused by cochlear implantation (CI) in congenital hearing loss patients. METHOD: Twenty-five patients were selected from a cohort of unilateral CI recipients to form the CI group. Static posturography was performed 1 to 3 days before and 3 to 5 days after CI. Each patient underwent the test with eyes open (EO) and eyes closed (EC) for 30 seconds, separately. Another group of age- and sex-matched patients with no history of hearing impairment undergoing unrelated surgeries formed the control group, and were examined with the same tests pre- and post-surgery. A third group of patients undergoing middle ear surgery formed the otitis media (OM) group. Postural sway parameters including sway velocity (SV) in the X-axis, SV in the Y-axis, length of sway locus length (LNG), and environmental area (ENV) were measured and recorded. RESULTS: Comparison of pre-surgery posturographical parameters between the CI and control groups revealed no significant differences. Significant differences were found in most parameters in pre- and post-surgery comparisons in the CI group. Mean SV values in the X-axis pre- and post-surgery were 8.48 and 11.52 mm/s, respectively, in the EO condition (P < .05), and 14.94 and 20.16 mm/s, respectively, in the EC condition (P < .05). In the Y-axis, mean SV values were 15.36 and 20.24 mm/s pre- and post-surgery, respectively, in the EC condition (P < .05). The LNG values in the CI group pre- and post-surgery were 319.60 and 469.88 mm in the EO condition (P < .05), and 571.40 and 764.12 mm in the EC condition (P < .05). No significant functional equilibrium change was observed in the control group between pre- and post-surgery (P > .05) except SV in the X-axis and LNG in the EO condition (P < .05). No significant pre- and post-surgery differences were found in the OM group. CONCLUSION: CI appeared to influence static equilibrium function within 1 week post-surgery. This influence was greater when eyes were closed.

Treatment for early postoperative esophageal fistula complicated with anterior cervical surgery.

OBJECT: To investigate the efficacy of debridement, open drainage, and early feeding to treat early esophageal fistula complicated with anterior cervical surgery. METHODS: Retrospective analysis was conducted on data from 3154 patients who had undergone anterior cervical surgeries for cervical vertebra diseases between January 2006 and January 2013, in which eight patients had esophageal fistula with five males and three females. Four patients had cervical spinal injuries and four patients had cervical spondylosis. All of whom had postoperative esophageal fistulas and underwent debridement and drainage. The wounds were left open for natural drainage. No drainage devices were placed in surgical sites, and no gastric tubes were placed after surgeries. Such normal food as rice balls but not liquid or semiliquid diet was recommended. Local pressurization was conducted with fists during feeding to prevent food overflowing from orificium fistulae. RESULTS: Healed esophageal fistulas were achieved in all of the patients after 1-2 weeks treatment. There weren't recurrent esophageal fistulas and delayed infection found during 2-5 years follow-up. Good swallowing functions and stable cervical vertebrae were achieved in all of the patients. The satisfactory therapeutic effects were achieved in patients with previous neck diseases. Frankel classifications were increased by 1-2 grades in patients with cervical spinal injuries. JOA scores were increased from 9.5 before surgery to 15.5 after surgery in patients with cervical vertebra diseases. CONCLUSIONS: Early postoperative esophageal fistula complication after anterior cervical surgery can successfully be treated by debridement, drainage without gastric tube, and with early oral postoperative feeding but without fluids.

Age-related changes in hypertensive brain damage in the hippocampi of spontaneously hypertensive rats.

The aim of the present study was to investigate the age­related alterations in hypertensive brain damage in the hippocampi of spontaneously hypertensive rats (SHR) and the underlying mechanisms. Aging resulted in a significant increase in the number of activated astrocytes and apoptotic cells in the SHR group, which was accompanied by increased expression of oxidative stress markers (iNOS and gp47phox) and apoptotic regulatory proteins (Bax and caspase­3). In addition, the expression of PPAR­Î³ and Bcl­2 were progressively reduced with increasing age in the SHR group. The 32 and 64­week­old SHRs exhibited significantly increased numbers of apoptotic cells, oxidative stress markers and pro­apoptotic proteins compared with age­matched WKY rats, which was accompanied by reduced expression of PPAR­Î³. Compared with the 16 and 32­week­old WKY group, the 64­week­old WKY rats exhibited increased oxidative stress and pro­apoptotic markers, and increased levels apoptotic cells. In conclusion, the present study indicated that both aging and hypertension enhanced brain damage and oxidative stress injury in the hippocampi of SHRs, indicated by an increased presence of apoptotic cells and astrocytes. In addition, reduced expression of PPAR­Î³ may contribute to the age­related brain damage in SHRs.

Elevated neuronal α-synuclein promotes microglia activation after spinal cord ischemic/reperfused injury.

The present study aimed to investigate the mechanism of injured neurons caused by ischemia/reperfusion in the induction of microglia activation. Spinal neurons were prepared and exposed to ischemic/reperfused conditions. The α-synuclein protein levels in these cells were analyzed by western blot, immunofluorescence, or enzyme-linked immunosorbent assay. Ischemia/reperfusion exposure led to elevated α-synuclein protein expression and release. Furthermore, when cocultured with injured neurons or supernatants from injured neurons, nitric oxide generation, H2O2 production, and tumor necrosis factor-α expression were promoted in microglia. Nevertheless, this effect was impeded by pretreatment of the α-synuclein antibody in the supernatants from injured neurons. Moreover, toll-like receptor 2 (TLR2) rather than TLR3 or TLR4 mediated microglia activation by α-synuclein. This process involved p38 MAPK and NF-κB activation, the inhibition of which resulted in reduced NADPH oxidase 2 (Nox2) in microglia. In conclusion, ischemia/reperfusion-injured neurons could express and release increased levels of α-synuclein and cause microglia activation through TLR2 both in vitro and in vivo.

Temperature-cycle microscopy reveals single-molecule conformational heterogeneity.

Our previous temperature-cycle study reported FRET transitions between different states on FRET-labeled polyprolines [Yuan et al., PCCP, 2011, 13, 1762]. The conformational origin of such transitions, however, was left open. In this work, we apply temperature-cycle microscopy of single FRET-labeled polyproline and dsDNA molecules and compare their responses to resolve the conformational origin of different FRET states. We observe different steady-state FRET distributions and different temperature-cycle responses in the two samples. Our temperature-cycle results on single molecules resemble the results in steady-state measurements but reveal a dark state which could not be observed otherwise. By comparing the timescales and probabilities of different FRET states in temperature-cycle traces, we assign the conformational heterogeneity reflected by different FRET states to linker dynamics, dye-chain and dye-dye interactions. The dark state and low-FRET state are likely due to dye-dye interactions at short separations.

[Influence of Nogo extracellular peptide residues 1-40 gene modification on survival and differentiation of neural stem cells after transplantation].

OBJECTIVE: To investigate the influence of Nogo extracellular peptide residues 1-40 (NEP1-40) gene modification on the survival and differentiation of the neural stem cells (NSCs) after transplantation. METHODS: NSCs were isolated from the cortex tissue of rat embryo at the age of 18 days and identified by Nestin immunofluorescence. The lentiviruses were transduced to NSCs to construct NEP1-40 gene modified NSCs. The spinal cords of 30 Sprague Dawley rats were hemisected at T9 level. The rats were randomly assigned to 3 groups: group B (spinal cord injury, SCI), group C (NSCs), and group D (NEP1-40 gene modified NSCs). Cell culture medium, NSCs, and NEP1-40 gene modified NSCs were transplanted into the lesion site in groups B, C, and D, respectively at 7 days after injury. An additional 10 rats served as sham-operation group (group A), which only received laminectomy. At 8 weeks of transplantation, the survival and differentiation of transplanted cells were detected with counting neurofilament 200 (NF-200), glial fibrillary acidic portein (GFAP), and myelin basic protein (MBP) positive cells via immunohistochemical method; the quantity of horseradish peroxidase (HRP) positive nerve fiber was detected via HRP neural tracer technology. RESULTS: At 8 weeks after transplantation, HRP nerve trace showed the number of HRP-positive nerve fibers of group A (85.17 +/- 6.97) was significantly more than that of group D (59.25 +/- 7.75), group C (33.58 +/- 5.47), and group B (12.17 +/- 2.79) (P < 0.01); the number of groups C and D were significantly higher than that of group B, and the number of group D was significantly higher than that of group C (P < 0.01). Immunofluorescent staining for Nestin showed no obvious fluorescence signal in group A, a few scattered fluorescent signal in group B, and strong fluorescence signal in groups C and D. The number of NF-200-positive cells and MBP integral absorbance value from high to low can be arranged as an order of group A, group D, group C, and group B (P < 0.05); the order of GFAP-positive cells from high to low was group B, group D, group C, and group A (P < 0.05); no significant difference was found in the percentage of NF-200, MBP, and GFAP-positive cells between group C and group D (P > 0.05). CONCLUSION: NEP1-40 gene modification can significantly improve the survival and differentiation of NSCs after transplantation, but has no induction on cell differentiation. It can provide a new idea and reliable experimental base for the study of NSCs transplantation for SCI.

The new role of LOX-1 in hypertension induced neuronal apoptosis.

Lectin-like oxidized low-density lipoprotein (oxLDL) receptor-1 (LOX-1) was originally identified as a receptor for oxLDL predominantly expressed in endothelial cells. Recently up-regulation of LOX-1 has been implicated in oxidative stress and cell apoptosis in many cell types. However, LOX-1 expression in neurons or regulation of neuronal apoptosis by LOX-1 has not been reported. To investigate the possible roles of LOX-1 in hypertension induced brain damage, we examined the distribution of LOX-1 in cortex and hippocampus and compared its expression in 32-week-old SHR and WKY rats. Immunofluorescence revealed that LOX-1 positive cells were located principally at the cortex involved in sensory information processing and were mainly expressed in neurons. We also found up-regulated mRNA expression of LOX-1, Bax and caspase-3 and down-regulated mRNA expression of Bcl-2 in SHR group. Compared with WKY group, SHR group showed increased LOX-1 positive cells and TUNEL positive cells. Furthermore, double-labeling method indicated that LOX-1 expression was colocalized with TUNEL positive cells, which means that LOX-1 expression was involved in hypertension related cell apoptosis. These findings indicated that LOX-1 expression was up-regulated in the cortex of SHR and its expression has implication in neuronal apoptosis. Elevated Bax/Bcl-2 ratio may be involved under this event.

[Construction and identification of Nogo extra cellular peptide residues 1-40 gene lentiviral vector].

OBJECTIVE: To construct a lentiviral expression vector carrying Nogo extra cellular peptide residues 1-40 (NEP1-40) and to obtain NEP1-40 efficient and stable expression in mammalian cells. METHODS: The DNA fragment of NEP1-40 coding sequence was amplified by PCR with designed primer from the cDNA library including NEP1-40 gene, and then subcloned into pGC-FU vector with in-fusion technique to generate the lentiviral expression vector, pGC-FU-NEP1-40. The positive clones were screened by PCR and the correct NEP1-40 was confirmed by sequencing. Recombinant lentiviruses were produced in 293T cells after the cotransfection of pGC-FU-NEP1-40, and packaging plasmids of pHelper 1.0 and pHelper 2.0. Green fluorescent protein (GFP) expression of infected 293T cells was observed to evaluate gene delivery efficiency. NEP1-40 protein expression in 293T cells was detected by Western blot. RESULTS: The lentiviral expression vector carrying NEP1-40 was successfully constructed by GFP observation, and NEP1-40 protein expression was detected in 293T cells by Western blot. CONCLUSION: The recombinant lentivirus pGC-FU-NEP1-40 is successfully constructed and it lays a foundation for further molecular function study of NEP 1-40.

Dietary mustard seeds (Sinapis alba Linn) suppress 1,2-dimethylhydrazine-induced immuno-imbalance and colonic carcinogenesis in rats.

In a Wistar rat model, prolonged supplementation of mustard seed (MS) to the diet significantly ameliorates the induction of colorectal carcinomas by 1,2-dimethylhydrazine (DMH). The expression of the splenocyte major histocompatibility complex class I (MHCI) was found significantly enhanced, whereas that of the major histocompatibility complex class II (MHCII) was significantly decreased. Compared to that of control animals, the proportion of spleenic B- and dendritic cells (DC) was amplified in the MS group. The expressions of MHCI, as well as that of MHCII, were increased in DC cells; whereas in B cells, MHCI expression was augmented but that of MHCII moderately decreased. The percentages of CD8+CD28+ and CD4+CD28+ cells were increased in the MS group, while the CD4+CD25+Foxp3+ subset was depressed. Plasma analysis showed that DMH-exposure induced amplified amounts of interleukin (IL)-4, IL-5, IL-10, and transforming growth factor-beta, whereas MS feeding counteracted this effect but enhanced IL-2, IL12p70, IL21, TNF-alpha, and interferon-gamma. In the SW480 colon adenocarcinoma cell-line, the cytotoxicity of spleenic T-cells from MS-fed animals was significantly increased. In the DMH-exposed rats, the expression of perforin in the spleenic T-cells was dramatically decreased, whereas MS abolished this depression. In summary, dietary MS suppresses DMH-induced immuno-imbalance as well as colon carcinogenesis in rats.