Upregulation of miR-335 reduces myocardial injury following myocardial infarction via targeting MAP3K2.

OBJECTIVE: Acute myocardial infarction (AMI) is a serious cardiovascular disease with a high incidence worldwide and the main cause of sudden cardiac death. The aim of this article was to study the protective role of miR-335 in myocardial infarction (MI) and the underlying molecular mechanism. MATERIALS AND METHODS: Thirty Sprague Dawley (SD) rats were randomly divided into sham group, MI + NC group and MI + agomiR-335 group. The expression of miR-335 in rat myocardium was detected by quantitative Real Time-Polymerase Chain Reaction (RT-PCR). Western blot was performed to detect the expression of TNF-α, IL-6, IL-1ß, Caspase-3, Cleaved Caspase-3 (C-Caspase-3) and MAP3K2 in rat myocardium. On the 7th day of the establishment of the rat MI model, a high-resolution small animal ultrasound system was utilized to detect the cardiac function of the rats, and the heart tissues and blood samples of the rats were collected. The corresponding kits were purchased to detect the contents of LDH, CK-MB, MDA and SOD in rat serum, and HE staining was employed to observe the morphology of rat myocardial tissue. RESULTS: The expression of miR-335 in myocardial infarcted zones and border zones of MI rats decreased significantly. The upregulation of miR-335 remarkably inhibited myocardial inflammation and apoptosis after MI, thus improving the cardiac function of MI rats. Compared with the sham group, the MAP3K2 expression in the MI + NC group was observably increased, while the overexpression of miR-335 could inhibit the expression of this protein. Through the Luciferase reporter gene experiment, we found that miR-335 could directly target MAP3K2. CONCLUSIONS: The expression of miR-335 decreased in myocardial tissue after MI, and the overexpression of miR-335 reduced myocardial damage by inhibiting oxidative stress, inflammation, and apoptosis via targeting MAP3K2, thereby improving the cardiac function of MI rats.

[Down-regulation of miR-146b-5p promotes malignant transformation of fusion cells after co-culture of macrophages with glioma stem cells in vitro].

Objective: To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms. Methods: Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform. RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot, fluorescence in situ hybridization, immunocytochemistry and chromosome karyotype analysis.The biological characteristics of fusion cells were further analyzed, together with relevant molecular changes. Results: RFP / EGFP double positive cells were obtained through in vitro co-culture. RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells. They also co-expressed GSCs marker Nestin and Mφ marker CD68, and karyotype analysis showed two types of characteristic chromosomes, which confirmed that the fusion cells originated from spontaneous fusion between SU4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate(18.83%) and lower tumor formation(4/5). Conclusion: Mφ could fuse with GSCs spontaneously in local tumor micro-environment. The proliferation and invasion abilities of fusion cells were higher than their parent cells, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas.

Determination of pseudocontact shifts of low-populated excited states by NMR chemical exchange saturation transfer.

Despite the critical roles of excited states in protein functions, they remain intractable for most structural studies because of their notably low populations. Chemical shifts for "invisible" states in slow exchange with the ground state are intuitively observed using nuclear magnetic resonance (NMR) chemical exchange saturation transfer (CEST) experiments. Here, we present a CEST NMR spectroscopy study for the observation of protein pseudocontact shifts (PCSs) of excited states, which carry valuable angular and distance information about these states. We first validate this approach, dubbed PCS-CEST, in the slow-exchange system of Abp1p SH3-Ark1p labeled with lanthanide ions, where the PCSs of the minor states resemble those of the holo-form ground state as expected. We further demonstrate that pre-existing folding transitional conformations of an FF domain exhibit remarkably lower PCS values than the ground state, which suggests that the low-populated ensemble is unfolded or largely unfolded. A higher resolution of PCSs of the minor states is achieved using our 1D selective CEST experiments. Thus, PCS-CEST provides an exquisite structural probe into the minor but functionally essential excited states.

Cloning and characterization of the SERK1 gene in triploid Pingyi Tiancha [Malus hupehensis (Pamp.) Rehd. var. pingyiensis Jiang] and a tetraploid hybrid strain.

This study aims to explore the roles of somatic embryogenesis receptor-like kinase (SERK) in Malus hupehensis (Pingyi Tiancha). The full-length sequences of SERK1 in triploid Pingyi Tiancha (3n) and a tetraploid hybrid strain 33# (4n) were cloned, sequenced, and designated as MhSERK1 and MhdSERK1, respectively. Multiple alignments of amino acid sequences were conducted to identify similarity between MhSERK1 and MhdSERK1 and SERK sequences in other species, and a neighbor-joining phylogenetic tree was constructed to elucidate their phylogenetic relations. Expression levels of MhSERK1 and MhdSERK1 in different tissues and developmental stages were investigated using quantitative real-time PCR. The coding sequence lengths of MhSERK1 and MhdSERK1 were 1899 bp (encoding 632 amino acids) and 1881 bp (encoding 626 amino acids), respectively. Sequence analysis demonstrated that MhSERK1 and MhdSERK1 display high similarity to SERKs in other species, with a conserved intron/exon structure that is unique to members of the SERK family. Additionally, the phylogenetic tree showed that MhSERK1 and MhdSERK1 clustered with orange CitSERK (93%). Furthermore, MhSERK1 and MhdSERK1 were mainly expressed in the reproductive organs, in particular the ovary. Their expression levels were highest in young flowers and they differed among different tissues and organs. Our results suggest that MhSERK1 and MhdSERK1 are related to plant reproduction, and that MhSERK1 is related to apomixis in triploid Pingyi Tiancha.

Antitumor efficacy of SarCNU in a human glioma xenograft model expressing both MGMT and extraneuronal monoamine transporter.

Treatment of malignant brain tumors with chloroethylnitrosoureas (CENUs) in addition to surgical resection and radiotherapy remains the foundation of glioma therapy. However, the clinical response to CENUs is at best modest. A novel analogue of nitrosoureas, 2-chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU), as compared to the standard CENU, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), has been demonstrated to have increased anticancer effects both in vitro and in vivo. Unfortunately, many human tumors have been known to be resistant to CENUs since they express DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT). In order to assess whether SarCNU has an effect on MGMT positive tumors, we evaluated its antitumor efficacy using an MGMT positive human glioma (SF-767) nude mouse xenograft model. Since SF-767 has high MGMT levels, BCNU treatment (20 mg/kg, Q4D x 3 i.p.) alone did not result in a satisfactory anticancer effect (p > 0.05). As expected, O6-benzylguanine (O6-BG) (100 mg/kg), which was given prior to BCNU treatment, by depleting MGMT activity, significantly enhanced BCNU antitumor efficacy (p < 0.001). Moreover, SarCNU treatment (167 mg/kg, Q4D x 3 i.p.) alone had a better antitumor effect than O6-BG plus BCNU treatment (F = 51.7, p = 0.0004). However, in this xenograft model, O6-BG did not significantly enhance the anticancer efficacy of SarCNU (F = 0.8, p = 0.411). The SF-767 human glioma xenograft is positive for extraneuronal monoamine transporter EMT (EMT) as determined by reverse-transcription polymerase chain reaction (RT-PCR). Our present results suggest that SarCNU is also effective for MGMT positive tumor if they exhibit EMT.

[Analysis of genetic diversity of Bama miniature pigs and Guizhou miniature pigs by RAPD].

RAPD analysis was performed with 31 selected single primers to study genetic diversity of two strains of miniature pigs, The percentages of polymorphisic loic in or between two strains of miniature pigs were 30.9%, 29.2% and 25.7% respectively, and the average genetic distances in and between two strains of miniature pigs were 0.120, 0.072, and 0.067 respectively. These results suggested that genetic diversity and genetic variability were poorer in and between two strains of miniature pigs than ordinary breeds' pigs.

Enhanced antitumor activity of sarCNU in comparison to BCNU in an extraneuronal monoamine transporter positive human glioma xenograft model.

A novel analogue of nitrosoureas, 2-chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU), has demonstrated increased anticancer effects in vitro and in vivo. Our previous work suggested that SarCNU enters cells via the extraneuronal monoamine transporter (EMT), that contributes to its enhanced cytotoxicity. In the present study, comparative activities of SarCNU and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) were evaluated in an EMT positive human glioma xenograft model. Athymic nude mice implanted subcutaneously or intracranially with human glioma SHG-44, a cell line that has been confirmed EMT positive by using reverse-transcription polymerase chain reaction (RT-PCR) assay, were treated with SarCNU at an optimal dose of 167 mg/kg, or BCNU at 20 mg/kg or 30 mg/kg, q4d x 3 intraperitoneally (i.p.). In 17 animals with subcutaneous tumor grafts treated with SarCNU, 9 animals became tumor free and 8 demonstrated tumor regression. While in the BCNU treated group, there were only 2 out of 10 mice in the 20 mg/kg group and 2 out of 7 in the 30 mg/kg group, which demonstrated some tumor regression. There were 4 drug related deaths in the BCNU (30 mg/kg) group, while there were no drug related deaths in the SarCNU group. In the intracranially implanted mice, the median survival time in the SarCNU group was more than 130 days, while in the BCNU treated group it was only 22 days which was similar to the control group (18 days). This is the first demonstration that SarCNU, in comparison to BCNU, has enhanced anticancer activity in an EMT positive human glioma xenograft model.

Cerebral sparganosis diagnosed and treated with stereotactic techniques. Report of two cases.

Infestation of the central nervous system with sparganum, the larva of Spirometra mansoni, is rare. Only 27 cases have been reported in the literature; however, the true incidence might be underestimated. Two cases are reported that were definitively diagnosed by stereotactic biopsy techniques. Without a positive histological diagnosis, the first case would have been diagnosed as having a metastatic brain tumor and a course of radiotherapy would probably have been initiated. Differentiation between granuloma of cerebral sparganosis and brain tumors such as meningiomas, gliomas, and metastatic tumors is often difficult before operation. Detailed magnetic resonance imaging might offer some help in such cases. Emphasis is placed on the increasing importance of stereotactic surgery in the diagnosis and treatment of an intracerebral mass lesion, including cerebral sparganosis.

Intramedullary spinal cord metastasis: report of a case and review of the literature.

Intramedullary spinal cord metastasis (ISM) is an unusual cause of myelopathy, in comparison to the common problem of extramedullary metastasis. Most patients with ISM are found at autopsy. When patients with ISM are diagnosed premorbidly, they usually have a previously known or concurrently diagnosable primary neoplasm. Radiation therapy is the definitive therapy. We recently encountered a patient who was seen initially because of progressive myelopathy, without evidence of underlying malignancy, in whom a papillary carcinoma was found at myelotomy. The condition improved after surgery and the patient returned to his previous work for five months. Subsequently, a lesion histologically identical with that obtained from the previous myelotomy was found in the lung. This case illustrates the value of surgical intervention in selected situation. When there is no identifiable organ metastasis or multiple intramedullary lesions, radical microsurgical resection of a focal intramedullary lesion should be considered.

Post-traumatic basal ganglia hemorrhage: analysis of 52 patients with emphasis on the final outcome.

A series of 52 patients suffering post-traumatic basal ganglia hemorrhage (BGH) after closed head injury is reviewed. Post-traumatic BGH was associated with other intracranial lesions in most cases. The outcome in this series is rather good, with 53.8% of patients having a functional survival. Radiologically, the post-traumatic intraventricular hemorrhage (IVH) and brainstem hemorrhage usually coexisted with a poor outcome. Old age (greater than 60), abnormal pupil reaction, impaired oculocephalic response, and abnormal motor response to pain stimuli are reliable clinical hallmarks for poor prognosis.

Spinal extradural cavernous hemangioma.

Three cases of isolated spinal extradural cavernous hemangiomas are reported, two in the thoracic and one in the lumbar region. One of them manifested as acute cord compression, the other two as chronic progressive myelopathy or radiculopathy. A total excision or subtotal excision with irradiation was performed. All of them had good functional recovery. The clinical picture, radiological diagnosis, and optimal method of treatment are discussed. The relevant literature is reviewed.

Carotid-cavernous sinus fistula associated with a primitive trigeminal artery.

Carotid-cavernous sinus fistulas are not rare, but they have never been reported in association with persistent primitive trigeminal artery. We recently encountered such a case. The Jaeger-Hamby procedure was employed, with mandatory occlusion of the primitive trigeminal artery.

Epistaxis due to traumatic intracavernous aneurysm: case report.

A patient who noted the onset of repetitive arterial epistaxis and progressive unilateral visual impairment 2 months after a blunt head injury was found to have a traumatic aneurysm of the cavernous portion of internal carotid artery (ICA). The epistaxis resolved and his vision recovered following a trapping procedure. This case and a review of previously reported cases emphasize the importance of early cerebral angiography in patients with post-traumatic epistaxis and visual failure.

The association of arteriovenous malformation and aneurysm within the posterior cranial fossa--report of a case.

A relatively rare case of cerebellar arteriovenous malformation (AVM) associated with a saccular aneurysm fed by the left posterior inferior cerebellar artery (PICA) is described. The interpretation of angiograms and the appropriate surgical treatment for the coexistence of the intracranial AVM and aneurysm are discussed. It is a fact that the steal phenomenon existed in the flow of contrast medium during the angiography. The appropriate surgical treatment may be clipping of the aneurysmal neck first and then radical excision of the AVM.

Upper lumbar disc herniation.

Herniations of the upper lumbar disc have often been overlooked due to a relatively low incidence we had 457 lumbar disc surgeries from January 1978 to June 1987. Twenty-two of them were of the upper three lumbar discs. We analyzed these patients according to their clinical syndromes, radiological findings and surgical results. The characteristic features of anterior thigh pain from upper lumbar disc herniation were different from the more commonly seen fourth and fifth lumbar disc herniation which resulted in sciatica. The incidence of cauda equina syndrome was relatively high in our series (36%). All the 22 cases had correlated X-ray findings. Satisfactory operative results for upper lumbar disc herniation were achieved in 20 of 22 (90%).

Acoustic neurinoma presenting as intratumoral bleeding.

The authors report a case of acoustic neurinoma presenting as intratumoral bleeding. This is the tenth reported occurrence. The literature is reviewed. All cases that have been reported have appeared with sudden onset of headache, vomiting, and decreased levels of consciousness. Aneurysm rupture, an arteriovenous malformation, or other vascular anomalies are suspected first. Preexisting unilateral hearing impairment is a valuable clue to differential diagnosis. Contrast-enhanced computed tomographic scans and cerebral angiograms are important tools for correct diagnosis. The tumor size (greater than 2 cm) and the thin, dilated vessels within the tumor are considered as pathogenetic factors for bleeding. When neurological status is not stable, placement of a ventriculoperitoneal shunt followed by urgent extirpation of the tumor is indicated.

Cystic meningioma--a case report.

A case of cystic meningioma is presented. The cyst in this particular case is within the tumor. Preoperatively, the diagnosis has been confused with astrocytoma, hemangioblastoma and neuroblastoma. Retrospectively, we conclude that careful evaluation of CT scan and angiogram will lead to accurate diagnosis.

Effects of temporary arterial occlusion on somatosensory evoked responses in aneurysm surgery.

Changes in the central conduction time (CCT) during the application of temporary clips were studied in 40 patients who had undergone operations for intracranial aneurysms in relation to postoperative neurological outcome. Ten of these 40 patients (25%) showed postoperative morbidity, although promptly recoverable in 5. None of the patients whose CCTs did not change following temporary occlusion of major vessels showed any postoperative morbidity, except in one case of anterior cerebral artery aneurysm. In 6 patients, temporary vascular occlusion caused a considerable transient prolongation in CCT of up to 10 msec. Two of these 6 patients were associated with postoperative neurological deficit (which was recoverable in 1). The cortical response became flat in 15 patients. Seven of these 15 patients showed hemispheric deficits postoperatively, although recoverable in 4. There was a correlation between the change in the somatosensory evoked response and postoperative outcome. Disappearance of the N20 potential following occlusion is regarded as a danger signal, but postoperative, irrecoverable neurological deficit seems to be unlikely if its disappearance takes more than 3-4 minutes. Even if the cortical response disappears, the clinical outcome is expected to be good if the N20 potential recovers within 20 minutes after recirculation.

The effects of halothane on somatosensory and flash visual evoked potentials during operations.

Intraoperative use of somatosensory evoked potentials (SEP's) to monitor intracranial aneurysm surgery and flash visual evoked potentials (F-VEP's) for parasellar surgery have been routinely employed in our clinic. We found that both EP modalities are sensitive to the changing concentration of our standard hypotensive agent, halothane. The prolongation of the N14-N20 interpeak latency to median nerve stimulation at the wrist, and prolongation of P100 latency with altered configuration of early VEP components to flash light stimulation, appear to be the results of direct pharmacological effects of the agent and not an effect of secondary hypotension. VEP is found easily abolished by halothane at a concentration of 2.0%, while the SEP is more resistant. Halothane is not ideal however when monitoring intraoperative VEP.

The application of flash visual evoked potentials during operations on the anterior visual pathways.

Flash visual evoked potentials (F-VEP's) have been recorded in fifteen patients (thirty eyes) with anterior visual pathway compression lesions before, during and after the operation in relation to clinical visual testing. A grading system for the wide variety of abnormal F-VEP's has been established, based on changes in forty patients (eighty eyes). No significant correlation between change in F-VEP and visual acuity or visual field has been demonstrated. Intraoperative F-VEP's have been found to be sensitive to halothane and to surgical manipulation of the optic nerve and/or chiasm during dissection of the tumour. The technique is considered a useful means of identifying the optic pathway during operation and may assist in prevention of optic damage if the transphenoidal approach is used.