MiR-29 inhibits neuronal apoptosis in rats with cerebral infarction through regulating Akt signaling pathway.

OBJECTIVE: The aim of this study was to explore the influence of micro ribonucleic acid (miR)-29 on neuronal apoptosis in rats with cerebral infarction by regulating the protein kinase B (Akt) signaling pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly divided into three groups, including: Sham group (n=12), Model group (n=12), and Inhibitor group (n=12). Common carotid artery, external carotid artery, and internal carotid artery were only exposed in the Sham group. However, the ischemia-reperfusion model was established by the suture method in the other two groups. After modeling, artificial cerebrospinal fluid was injected into the lateral ventricle in the rats of the Sham and Model groups. Similarly, miR-29 inhibitor was injected into the lateral ventricle in the rats of the Inhibitor group. At 24 h postoperatively, the sampling was performed. Zea-Longa score was used to evaluate the neurological deficit of rats. Meanwhile, the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2 associated X protein (Bax) in cerebral tissues were detected via immunohistochemistry. The protein expression levels of Akt and phosphorylated Akt (p-Akt) were determined using Western blotting. Furthermore, the expression of miR-29 and cell apoptosis were detected via quantitative Polymerase Chain Reaction (qPCR) and terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay, respectively. RESULTS: Compared with Sham group, Model, and Inhibitor groups had substantially raised the Zea-Longa scores (p<0.05). The Zea-Longa score in the Model group was markedly lower than that of the Inhibitor group (p<0.05). The positive expression level of Bax was remarkably upregulated (p<0.05). However, the positive expression level of Bcl-2 declined dramatically in both Model group and Inhibitor group when compared with the Sham group (p<0.05). Besides, the Model group exhibited significantly lower positive expression level of Bax and higher positive expression level of Bcl-2 than the Inhibitor group (p<0.05). The relative protein expression level of p-Akt markedly increased in the Model and Inhibitor groups when compared with the Sham group (p<0.05). However, it was considerably higher in the Model group than that of the Inhibitor group (p<0.05). In comparison with the Sham group, both Model group and Inhibitor group exerted substantially elevated expression level of miR-29 (p<0.05). The relative expression level of miR-29 in the Model group was significantly upregulated when compared with the Inhibitor group (p<0.05). The apoptosis rate of cells in both Model group and Inhibitor group was markedly higher than that of the Sham group (p<0.05). Furthermore, the Model group showed remarkably lower apoptosis rate than the Inhibitor group (p<0.05). CONCLUSIONS: MiR-29 inhibits neuronal apoptosis in cerebral infarction rats by upregulating the Akt signaling pathway, thereby serving as a protector.

Efficacy and safety of minimal pedicle screw fixation for thoracolumbar fractures: a meta-analysis.

OBJECTIVE: Minimal and open pedicle screw fixation procedures have been widely used in the treatment of thoracolumbar fractures. However, the efficacy and safety of these approaches remain unclear. This meta-analysis was conducted to evaluate perioperative, functional and radiological outcomes of percutaneous versus open pedicle screw fixation for thoracolumbar fractures. MATERIALS AND METHODS: To obtain relevant literature, a systematic search was performed using the MEDLINE, EMBASE, and Cochrane databases. The Cowley criteria were used to evaluate the risk of bias for the included studies. A database that included patient demographic information and perioperative outcomes was established. Summary odds ratios (ORs) and weighted mean differences (WMDs) with 95% confidence intervals (CIs) were estimated. Analyses were performed for the two subgroups of Chinese studies and studies from other nations. Publication bias was assessed using the funnel plot method. RESULTS: Eleven comparative observational studies that satisfied our inclusion criteria were identified via a literature search in the MEDLINE, EMBASE, and Cochrane databases. Relative to the open approach, the minimal approach was associated with less blood loss (WMD=-218.10, 95% CI: -266.31 to -169.88, p<0.00001) and shorter operative time (WMD=-15.31, 95% CI: -24.73 to -5.88, p=0.001). Evidence indicated that a significant difference was observed between Chinese studies and other studies with respect to blood loss (p=0.02). We also found that the minimal approach was associated with a lower postoperative visual analog scale (VAS) score (WMD = -1.06, 95% CI: -1.32 to -0.8, p<0.00001) and less correction loss (WMD=-0.59, 95% CI: -1.16 to 0.02, p=0.04) than the traditional open approach. No significant difference between these approaches was found with respect to complication rate (OR 0.78, 95% CI: 0.39 to 1.55, p=0.48). CONCLUSIONS: The evidence indicated that the minimal approach had better functional and radiological outcomes than the open approach. Neither approach was superior with respect to complication rate. Relative to the open approach, the minimal approach might be associated with decreased operative time, less blood loss and a shorter hospital stay.

[Establishment and evaluation of a new method for determining hemodynamics of pulmonary hypertension rats].

Objective: By evaluating the hemodynamic parameters such as cardiac output (CO), right ventricular pressure (RVP), pulmonary artery pressure (PAP) and total pulmonary resistance index (TPRI) in pulmonary hypertension rat model, we established a more comprehensive hemodynamic evaluation system, which objectively evaluated the severity of disease and exercise tolerance in rats with pulmonary hypertension. Methods: SD rats were randomly divided into a control group and a model group with 5 rats in each group. The model group was intraperitoneally injected with SU5416 (20 mg/kg) and placed in an oxygen chamber at a 10% oxygen concentration for 21 days and then placed in a normoxic environment for 14 days. After modeling, rats were anesthetized and mechanically ventilated. The operator cut the skin along the right paraxial line, detached and ligated the intercostal artery, and then cut off the 3 and 4 ribs, exposing the heart and freeing aortic root about 0.2 cm. The flowmeter probe was set in the dissected aortic segment, and real-time recording time, blood flow waveforms, cardiac output were calculated accordingly. Then the needle attached to the baroreceptor was inserted into the right ventricle and the system acquired the right ventricular time-pressure waveform. After the waveform stabilized for about 30 seconds, the end of the cannula was sent to the pulmonary artery trunk through the entrance of the pulmonary artery to record the time-pressure curve of the pulmonary artery. Results: RVSP, PASP, PADP and mPAP in the model group were significantly higher than those of the control group [ RVSP(23.4±5.4) mmHg, 1 mmHg=0.133 kPa vs (56.4±13.0) mmHg, PASP (22.8±4.4) mmHg vs (58.5±14.9) mmHg, PADP (9.7±1.9) mmHg vs (30.3±7.0) mmHg, mPAP (14.1±2.7) mmHg vs (41.9±8.0) mmHg, all P<0.05 ]. Compared with the control group, the cardiac index in the model group was significantly lower [ CI (0.54±0.08) ml·min(-1)·g(-1) vs (0.40±0.09) ml·min(-1)·g(-1,) P=0.02 ]. Furthermore, compared with the control group, pulmonary vascular resistance index was significantly increased in the model group[PVRI (0.27±0.03) mmHg·ml(-1)·min(-1)·kg(-1) vs (0.06±0.01) mmHg·ml(-1)·min(-1)·kg(-1,) P<0.05]. The pathological results also showed that the middle part of pulmonary arterioles in the model group had muscular hypertrophy and muscular pulmonary arterioles, and even plexiform lesions. Conclusion: In this study, we established a new method that simultaneously determined several hemodynamic parameters such as RVSP, PASP, PADP, CO, CI and PVRI, which provided a more comprehensive assessment of hemodynamic changes in pulmonary hypertension rat models.

The embryonic and evolutionary boundaries between notochord and cartilage: a new look at nucleus pulposus-specific markers.

The adult nucleus pulposus (NP) and articular cartilage are similar in terms of their histocytological components and biomechanical functionalities, requiring a deep understanding of NP-specific markers to better evaluate stem-cell-based NP regeneration. Here, we seek to distinguish NP cells from articular chondrocytes (ACs), focusing on differences in their embryonic formation and evolutionary origin. Embryonically, NP cells are conservatively derived from the axial notochord, whereas ACs originate in a diversified manner from paraxial mesoderm and neural crest cells. Evolutionarily, although the origins of vertebrate NP and AC cells can be traced to similar structures within protostomia-like bilaterian ancestors, the distant phylogenetic relationship between the two groups of animals and the differences in the bodily origins of the tissues suggest that the tissues may in fact have undergone parallel evolution within the protostomia and deuterostomia. The numbers of supposedly NP-specific markers are increasing gradually as microarray studies proceed, but no final consensus has been attained on the specificity and physiology of "exclusive" NP markers because of innate variations among species; intrinsic expression of genes that destabilize the circadian clock; and cooperation by, and crosstalk among, different genes in terms of physiology-related phenotypes. We highlight the embryonic and evolutionary boundaries between NP and AC cells, to aid in recognition of the challenges associated with evaluation of the role played by nucleopulpogenic differentiation during stem-cell-based intervertebral disc regeneration.

Drug-induced hypersensitivity syndrome caused by valproic acid as a monotherapy for epilepsy: First case report in Asian population.

Valproic acid (VPA) is a broad-spectrum antiseizure drug used for a variety of clinical conditions, such as epilepsy and mood disorders. Drug-induced hypersensitivity syndrome (DRESS) accompanied by hyponatremia, thrombocytopenia, hypoalbuminemia and elevated aminotransferase has never been reported as an adverse effect of VPA monotherapy during titration for epilepsy in Asian population. Hereby, we present the case of a 73-year-old Chinese male who suffered from DRESS and other complications two weeks after initiating VPA treatment for epilepsy. Understanding the risk associated with VPA-induced DRESS, and taking effective measures to avoid the severe side effects are necessary.

[Method for recording tension changes of small pulmonary artery and vein using the isolated vessel tension measurement system].

Objective: To establish a standardized method for isolated pulmonary artery and vein rings with different diameter, pressure and length, which could provide a more scientific method for in vitro study of pulmonary vessel diseases. Methods: Male SD rats were anesthetized, and the right ventricular systolic pressure were measured. Small pulmonary artery and vein rings with 200-400 µm in diameter and 2 mm in length were prepared by dissecting pulmonary arteries and veins. The pulmonary vessel rings were mounted in the organ bath by 2 stainless steel wires with diameter of 40 µm. Then the internal circumference of the vessel rings was increased gradually with 100 µm per step. At the same time the vascular tension was recorded by the Myograph System and Acknowledgement data acquisition system, and subsequently the passive length-tension exponential curve was made. The initial tension of the rings was set, equilibrated for another 30 min, and then stimulated with 60 mmol/L KCl 3 times, and the best contractile reactivity was achieved. The contractile reactivity of pulmonary artery rings and endothelial integrity were detected by exposure to 1 µmol/L phenylephrine(PE) and 10 µmol/L acetylcholine(Ach), while the contractile reactivity of pulmonary vein rings was detected by exposure to 1 µmol/L U46619 and 10 µmol/L papaverine. Results: The contraction and relaxation effects of the pulmonary artery rings reached 0.39 mg and 92% when they were stimulated by 1 µmol/L PE and 10 µmol/L Ach. The contraction and relaxation effects of pulmonary vein rings were up to 0.13 mg and 84% when they were exposed to 1 µmol/L U46619 and 10 µmol/L papaverine, respectively. Conclusion: Pulmonary artery and vein rings with appropriate basal tension and optimal vasodilator activity were prepared, and a standardized method of tension experiment for isolated pulmonary artery and vein rings established.

Apparent First-Order Wetting and Anomalous Scaling in the Two-Dimensional Ising Model.

The global phase diagram of wetting in the two-dimensional Ising model is obtained through the exact calculation of the surface excess free energy. In addition to a surface field for inducing wetting, a surface-coupling enhancement is also included. The wetting transition (of second order) is critical for any finite ratio of surface coupling J_{s} to bulk coupling J, and becomes of first order in the limit J_{s}/J→∞. However, for J_{s}/J≫1, the critical region is exponentially small and is practically invisible to numerical studies. A distinct preasymptotic regime exists in which the transition displays first-order character. In this regime, surprisingly, the surface susceptibility and surface specific heat develop a divergence and show anomalous scaling with an exponent equal to 3/2.

Hypoxia regulates sumoylation pathways in intervertebral disc cells: implications for hypoxic adaptations.

OBJECTIVE: To explore the hypoxic regulation of sumoylation pathways and cell viability in nucleus pulposus (NP) and annulus fibrosus (AF) cells. DESIGN: Expression of small ubiquitin-like modifier (SUMO) molecules, SUMO E1 activating enzymes SAE1 and SAE2, SUMO E2 conjugating enzyme UBC9, and de-sumoylation enzyme sentrin/SUMO-specific proteases (SENP)1 was immunolocalized in rat intervertebral disc (IVD) cells. NP and AF cells were cultured in hypoxia and cell viability was evaluated by quantifying cell proliferation, cellular senescence, apoptosis, and cell cycle distribution. Hypoxic regulation of sumoylation pathways was studied by analyzing the transcription and expression of SUMO molecules and sumoylation enzymes. Loss of function study using SENP1 siRNA was performed to investigate the regulatory role of sumoylation on the function of hypoxia inducible factor 1α (HIF-1α) and the hypoxic tolerance of IVD cells. RESULTS: Sumoylation pathways were expressed in IVD cells and localized predominantly in nuclei. Both NP and AF cells maintained viability under hypoxia and upregulated the expression of SENP1. In NP cells hypoxia transiently increased the expression of SUMO-1, SUMO-2/3, SAE2, and UBC9, whereas SUMO-1 was elevated while SUMO-2/3, SAE1, SAE2, and UBC9 were reduced by low oxygen tensions in AF cells. Although downregulation of SENP1 decreased the transcriptional activity of HIF-1α, the viability of disc cells showed no significant loss under hypoxia. CONCLUSIONS: NP and AF cells equally tolerate oxygen deficiency, but differently regulate the sumoylation pathways under hypoxia. The distinct sumoylation dynamics may help extend our understanding of the cell-specific regulation of the molecular basis that promotes cell survival in the hypoxic IVD.

Aging and age related stresses: a senescence mechanism of intervertebral disc degeneration.

Intervertebral disc (IVD) degeneration is a complicated process that involves both age-related change and tissue damage caused by multiple stresses. In a degenerative IVD, cellular senescence accumulates and is associated with reduced proliferation, compromised self-repair, increased inflammatory response, and enhanced catabolic metabolism. In this review, we decipher the senescence mechanism of IVD degeneration (IVDD) by interpreting how aging coordinates with age-related, microenvironment-derived stresses in promoting disc cell senescence and accelerating IVDD. After chronic and prolonged replication, cell senescence may occur as a natural part of the disc aging process, but can potentially be accelerated by growth factor deficiency, oxidative accumulation, and inflammatory irritation. While acute disc injury, excessive mechanical overloading, diabetes, and chronic tobacco smoking contribute to the amplification of senescence-inducing stresses, the avascular nature of IVD impairs the immune-clearance of the senescent disc cells, which accumulate in cell clusters, demonstrate inflammatory and catabolic phenotypes, deteriorate disc microenvironment, and accelerate IVDD. Anti-senescence strategies, including telomerase transduction, supply of growth factors, and blocking cell cycle inhibitors, have been shown to be feasible in rescuing disc cells from early senescence, but their efficiency for disc regeneration requires more in vivo validations. Guidelines dedicated to avoiding or alleviating senescence-inducing stresses might decelerate cellular senescence and benefit patients with IVD degenerative diseases.

Unravelling the multifaceted roles of Atg proteins to improve cancer therapy.

Autophagy follows a lysosomal degradation pathway in which a cell digests its own components. It is highly regulated by a limited number of autophagy-related genes (Atg) and the proteins they encode, that are crucial for cells to undergo the process via modulating autophagsome formation. Recently, accumulating evidence has revealed the core molecular machinery of autophagy; however, intricate relationships between autophagy and cancer remain an enigma. Several studies have shown that Atgs can play an important role in carcinogenesis, by which Atgs may modulate a series of oncogenic and tumour suppressive pathways, implicating microRNA (miRNA) involvement. In this review, we will present the key role of Atgs in deciding the fate of cancer cells, discuss some representative Atgs and their proteins such as ULK, Beclin-1, and Atg8/LC3-Atg4, which can also be regulated by miRNAs. Thus, Atgs can be considered to be targets for cancer treatment, which may illuminate the future of cancer therapy.

Interictal magnetoencephalography used in magnetic resonance imaging-negative patients with epilepsy.

PURPOSE: This study aims to investigate the contributions of magnetoencephalography (MEG) in magnetic resonance imaging (MRI)-negative patients. METHODS: A total of 18 MRI-negative patients diagnosed with refractory epilepsy, subjected to MEG investigation, and subsequently underwent surgery were selected for retrospective analysis. A 1.5-tesla Magnetom Sonata with an eight-channel head array coil was used. MEG data were obtained using a 74/248-channel system. RESULTS: A total of 16 patients (16/18) had positive MEG results, comprising 12 patients with monofocal localizations, five with multifocal localizations, and one with unremarkable results in MEG. In addition, 12 patients had indicative single photon-emission computed tomography (SPECT), five had indicative fluorodeoxyglucose positron emission tomography (FDG-PET), and all the patients had intracranial electroencephalography (EEG) (14 with subdural electrodes and four with electrocorticography). The intracranial EEG recordings of nine patients were guided by MEG informative results. Among these 18 patients, 10 exhibited good postoperative outcomes (Engel I and II), four of which were completely seizure-free. All these ten patients had clear monofocal localization in MEG, including nine with accordant indicative metabolic changes in either SPECT or FDG-PET, or both. None of the five patients with multifocal localizations achieved good postoperative outcomes. CONCLUSION: For cases with negative MRI findings, epilepsy surgery may be an alternative option for pharmaco-resistant patients if epileptogenic focus localizations by MEG are present in multimodal evaluation.

Isolation and characterization of Aeromonas schubertii from diseased snakehead, Channa maculata (Lacepède).

Pure bacterial cultures were isolated from diseased snakeheads, Channa maculata (Lacepède), suffering high mortality in a farm in Zhongshan, southern China. Three isolates, namely ZS20100725, ZS20100725-1 and ZS20100725-2, were identified as Aeromonas schubertii. All the isolates showed high 16S rRNA sequence similarities with A. schubertii. The isolates exhibited strong virulence to snakeheads in experimental challenges with LD(50) ranging between 1.4 × 10(4) and 6.4 × 10(6) CFU g(-1). Two of the isolates were positive for haemolysin, elastase, lipase and lecithinase by phenotypic determination, which was further confirmed by PCR amplification of the haemolysin and elastase genes. In sterile liquid medium, the best growth conditions of strain ZS20100725 were 30 °C, pH 7 and 0.5% salinity (w/v). Antibiotic susceptibility tests showed that strain ZS20100725 was susceptible to cefoxitin, cefoperazone and chloramphenicol. Furthermore, histopathology of diseased snakeheads infected with A. schubertii showed necrosis and congestion in liver, kidney and spleen and also damage to the cardiac muscle, intestine and gills.

Homogeneous and inhomogeneous phase transitions in the Blume-Capel model with random bonds.

Using mean field approximation, we investigate the Blume-Capel model with random bond. The system is self-organized into blocks, which behave like superspins and are coupled with their neighbors. The methods to search the elementary blocks and calculate their coupling are proposed. For the strongly disordered case, the coupling distribution is exponential and the phase transition should be inhomogeneous. This physical picture should be common in various phase transition in quenched disordered systems.

Association between carbamazepine-induced cutaneous adverse drug reactions and the HLA-B*1502 allele among patients in central China.

The aim of this study was to investigate the association between carbamazepine (CBZ)-induced cutaneous adverse drug reactions (cADRs) and the HLA-B*1502 allele among patients from central China. Eight patients with Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN), 28 with mild maculopapular eruptions (MPEs), 50 CBZ-tolerant controls, and 71 healthy volunteers were recruited. HLA genotyping was performed using the polymerase chain reaction sequence-based typing (SBT) method. As a result, the HLA-B*1502 allele was observed at the following rates: (1) 100% (8/8) among those with CBZ-induced SJS/TEN, (2) 10.7% (3/28) among those with CBZ-induced MPEs; (3) 8.0% (4/50) among CBZ-tolerant controls; (4) 8.5% (6/71) among healthy volunteers. The eight patients with SJS/TEN positive for the HLA-B*1502 allele had an odds ratio (OR) of 184 compared with CBZ-tolerant controls. There was no significant difference in frequency between patients with MPEs and CBZ-tolerant controls (P>0.05). Thus, CBZ-induced SJS/TEN, but not MPEs, is strongly associated with HLA-B*1502. Testing for HLA-B*1502 should be recommended for patients from central China prior to initial CBZ treatment.

A preliminary study on the feasibility of gene expression profile of rhesus monkey detected with human microarray.

OBJECTIVES: A pig-to-monkey transplant model was initiated to investigate the outcome of pig-to-human xenotransplantation. Though monkey is close to human in biology and physiology, the genetic differences between the two species remains unclear. This study sought to compare the gene expressions of three tissues from humans and rhesus monkey. METHODS: RNA samples extracted from liver, spleen, and peripheral blood cells were hybridized onto Illumina gene expression microarray. Genes with detected signals greater than 1000 and diff-scores higher than 100 were selected as significant results. The data were analyzed with Illumina software. mRNA expression levels were confirmed by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis. RESULTS: Of the 47,293 transcripts tested on every gene chip, more than 6000 genes were expressed in three tissues. Total numbers of genes detected and the similarity ratios followed the same rule as liver < PBC < spleen. The 136 IRI-related genes, 192 immunological-related genes, and 131 cell cycle-related genes selected and analyzed showed gene expression concordance rates of 82.35%, 72.92%, and 77.10%, respectively. RT-PCR tests indicated similar mRNA expression levels of RTN4, interleukin (IL)-1beta, NF-kappaB1, IL-8, and G0S2 to the results on chips. CONCLUSIONS: The detected mRNA expressions in human and monkey tissues showed an average consistency in 85.78%, indicating that a human microarray might provide a part of the information for monkey sample testing. Therefore, in pig-to-monkey transplant models, monkey microarray may be used to determine recipient gene expressions. The genetic difference between human and monkey must be taken into account in interpreting the experimental results.

Interpenetration in [Cd(isonicotinate)2(1,2-bis(4-pyridyl)-ethane)0.5(H2O)]n, a novel octahedral polymer containing an unusual two-dimensional bilayer motif generated by self-assembly of rectangle building blocks.

A hydrothermally synthesized mixed-ligand molecular solid composed of pillared bilayers has been formulated; it presents a new remarkable three-dimensional architecture sustained by the catenation of two-fold decked layers, and exhibits intense photoluminescence upon photoexcitation at 314 nm.

High resolution spectral analysis of 13CH3OH in the excited torsional states.

In this work, we have extended our previous analysis of the Hamiltonian of 13C substituted methanol to include a large number of spectral lines involving the second excited torsional state using an improved model. The data set consisted of 2529 Fourier transform and microwave transitions with the rotational angular momentum J < or = 10, K < or = 6 and n < or = 2 (with 336 MW lines). The data set was fitted with the new Hamiltonian model to derive the molecular parameters. The results indicate that the model developed for the other methanol species (CH3OH, CH3(18)OH and CH3OD) is also valid for the C-13 substituted species. The results will allow the energy levels of the molecule to be calculated for higher torsional levels above the internal rotational barrier with improved precision and allow the analysis to be carried out for more excited torsional states.

[An experimental study and preliminary clinical report of shape-memory sawtooth-arm embracing internal fixator].

The sawtooth-arm internal embracing fixator is made of Nickel-Titanium shape memory alloy and consists of 3 components: body, arms and sawteeth. In cross section a 2/3 circumference is constructed by body and arms of the fixator. The free ends of the arms which exceed the semi-circle are bent more medially so that it can match the requirement of fixation of long tubular bone whose cross section is not a regular circle. The embracing fixator has two types: the cylinder type is for use in the middle part and the cone type is either for the proximal or distal 1/3 of the shaft of long tubular bone. The animal experimental studies and in vitro mechanical tests demonstrated that the embracing fixator possesses good anti-bending and anti-torsion effects, and its resisting compression effect was much lower than that of bone plate. These characteristics are beneficial for enhancing fracture healing, reducing post fixation osteoporosis and providing a new, simple and effective method for the treatment of long tubular bone fracture. The fixator has already been used in the treatment of fractures of femur, humerus, radius and ulna with good results.