Establishment of a screening model for compounds uncoupling the interaction of nNOS with PSD-95 / 药学学报

In ischemic stroke, increased level of neuronal complex of nitric oxide synthase (nNOS)-postsynaptic density protein-95 (PSD-95) plays an important role in neuronal damage. We aimed to establish a screening model to identify compounds capable of uncoupling nNOS interaction with PSD-95. In this model, human embryonic kidney-293T (HEK-293T) cells were transfected with either pCDH-Flag-nNOS or pcDNA3.1-PSD-95 plasmid to obtain the protein of Flag-nNOS or PSD-95. Incubating Flag-nNOS with PSD-95 causes formation of the nNOS-PSD-95 complex. ZL006, a known uncoupler of nNOS-PSD-95 interaction, can disturb the interaction between Flag-nNOS and PSD-95, serving as a positive control. The method coupling antibodies to magnetic beads with glutaraldehyde was used to decrease the cost and increase the efficiency. To establish that our model is suitable for selecting nNOS-PSD-95 uncouplers, we evaluated the ability of IC87201, another reported uncoupler of nNOS-PSD-95 interaction, and structural analogs of ZL006. IC87201 and one structure analog of ZL006 showed uncoupling effect, supporting that our model can be used to select different types uncoupler blocking nNOS-PSD-95 interaction.

Short-term Preoperative Octreotide for Thyrotropin-secreting Pituitary Adenoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Thyrotropin-secreting pituitary adenomas (TSHomas) are a rare cause of hyperthyroidism. Somatostatin (SST) analogs work by interacting with somatostatin receptors (SSTRs). This study aimed to evaluate short-term preoperative octreotide (OCT) use in TSHoma patients and to investigate SSTR2 and SSTR5 expression and observe structural changes in tumor tissue.</p><p><b>METHODS</b>We reviewed records and samples from eight TSHoma patients treated between July 2012 and July 2015. We tested immunohistochemically for SSTR2/5 expression and examined TSHoma cells for morphological changes. Signed rank sum test was used to compare the efficacy of short-term preoperative OCT treatment.</p><p><b>RESULTS</b>OCT treatment (median time: 7.9 days, range: 3-16 days; median total dose: 1.8 mg, range: 0.9-4.2 mg) led to significant decrease in all patients' thyroid hormone levels (FT3 [nmol/L]: 8.33 [7.02, 12.29] to 4.67 [3.52, 5.37] [P = 0.008]; FT4 [pmol/L]: 25.36 [21.34, 28.99] to 16.66 [14.88, 21.49] [P = 0.016]; and TSH [μU/ml]: 5.80 [4.37, 6.78] to 0.57 [0.19, 1.24] [P = 0.008]). All the eight tumor specimens expressed high SSTR2 protein levels; 5/8 expressed high SSTR5, but 3/8 that expressed low SSTR5 presented a significantly higher TSH suppression rate (P = 0.036). Electron microscopy showed subcellular level impairments, including clumped nuclear chromatin and reduced cytoplasmic volume. Golgi complexes were observed in the OCT-treated TSHoma specimens.</p><p><b>CONCLUSIONS</b>OCT can control hormone levels and damage the ultrastructure of tumor cells and organelles. Short-term response to OCT may be related to SSTR5 expression. Preoperative SST analog treatment for TSHoma could be considered as a combination therapy.</p>

Endoscopic treatment of cerebral hemispheres convex arachnoid cysts / 中华神经医学杂志

Objective To investigate and evaluate the effectiveness ofneuroendoscopic therapy on patients with cerebral hemispheres convex arachnoid cysts.Methods Thirteen patients with cerebral hemispheres convex arachnoid cysts (close to lateral ventricle or cistern),admitted to and accepted endoscopic cystoventriculostomy or cystocisternotomyin our hospital from September 2007 to July 2011,were chosen in our study; their clinical data,including clinical and neuroradiological presentations,surgical indications,surgical techniques,complications,and clinical and neuroradiological follow-up results,were retrospectively analyzed.Results The cysts were reduced in size in all 13 patients (100％).The symptoms disappeared in 9 and improved in 4 after the operation,with a clinical improvement rate reaching 100％.One patient had asymptomatic subdural hydroma after the operation.Conclusion Endoscopic fenestration is an effective treatment for patients with cerebral hemispheres convex arachnoid cysts close to lateral ventricle or cistern,and can be the first surgical choice for these cysts.

Clinical observation on nerve growth factor in the treatment of severe craniocerebral trauma / 中华神经医学杂志

Objective To investigate the influence of nerve growth factor (NGF) on the level of plasma creatine kinase BB (CK-BB) and its clinical effect in patients with severe craniocerebral trauma. Methods Eighty patients with severe craniocerebral trauma (GCS≤8) were randomly assigned to NGF-treated (n=40) and control (n=40) groups. Conventional therapy was performed on these patients and NGF was added into the NGF-treated group. The level of plasma CK-BB in these patients was measured; the clinical prognosis of the patients was evaluated based on the GOS scores 6 months after severe craniocerebral trauma; the condition of regained consciousness was compared between the 2 groups. Results Plasma CK-BB in the NGF-treated group was significantly lower than that in the control group (P<0.05); the recovery rate in the NGF-treated group (24/40) was significantly higher than that in the control group (11/40); the consciousness regained within i month was found in 32 patients in the NGF-treated group but only 18 of that was noted in the control group (P<0.05); NGF-treated group (12.43±6.25) showed a significantly shorter time that consciousness was regained as compared with the control group (15.96±7.58). Conclusion NGF can decrease the level of plasma CK-BB, shorten coma time and improve life quality in patients with severe craniocerebral trauma. Application ofNGF, a safe and effective treatment method, provides a new potential therapeutic strategy for patients with CNS injury.

Endoscopic treatment of middle fossa arachnoid cysts / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate and evaluate the effectiveness of neuroendoscopic therapy for arachnoid cysts of middle cranial fossa.</p><p><b>METHODS</b>From January 2004 to June 2009, 32 patients with arachnoid cysts of middle cranial fossa who were treated with endoscopic cystocisternal fenestration were retrospectively analyzed. There were 21 male patients and 11 female patients, aged from 6 months to 39 years. The clinical and neuroradiological presentation, indications, surgical technique, complications, and clinical and neuroradiological follow-up were analyzed.</p><p><b>RESULTS</b>The cysts were reduced in size in 20 patients and completely disappeared in 4 patients. For the 27 patients with symptoms before operation, the symptoms disappeared in 8 cases and improved in 17 cases after operation. There were asymptomatic subdural hydroma in 4 patients, intracranial infection and incision cerebro-spinal fluid leakage in 1 patient respectively. The complication incidence rate was 18.8%.</p><p><b>CONCLUSIONS</b>Endoscopic fenestration is an effective treatment for symptomatic arachnoid cysts of middle cranial fossa and could be performed as the first surgical choice for these patients.</p>

Differential gene expression by fiber-optic beadarray and pathway in adrenocorticotrophin-secreting pituitary adenomas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Adrenocorticotrophin (ACTH)-secreting pituitary adenomas account for approximately 7% - 14% of all pituitary adenomas, but its pathogenesis is still enigmatic. This study aimed to explore mechanisms underlying the pathogenesis of ACTH-secreting pituitary adenomas.</p><p><b>METHODS</b>We used fiber-optic beadarray to examine gene expression in three ACTH-secreting adenomas compared with three normal pituitaries. Four differentially expressed genes from the three ACTH-secreting adenomas and three normal pituitaries were chosen randomly for validation by reverse transcriptase-real time quantitative polymerase chain reaction (RT-qPCR). We then analyzed the differentially expressed gene profile with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway.</p><p><b>RESULTS</b>Fiber-optic beadarray analysis showed that the expression of 28 genes and 8 expressed sequence tags (ESTs) were significantly increased and the expression of 412 genes and 31 ESTs were significantly decreased. Bioinformatic and pathway analysis showed that the genes HIGD1B, EPS8, HPGD, DAPK2, and IGFBP3 and the transforming growth factor (TGF)-β signaling pathway and extracellular matrix (ECM)-receptor interaction pathway may play important roles in tumorigenesis and progression of ACTH-secreting pituitary adenomas.</p><p><b>CONCLUSIONS</b>Our data suggest that numerous aberrantly expressed genes and several pathways are involved in the pathogenesis of ACTH-secreting pituitary adenomas. Fiber-optic beadarray combined with pathway analysis of differential gene expression appears to be a valid method of investigating tumour pathogenesis.</p>

Establishment of intramedullary spinal cord glioma model in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Treating intramedullary spinal cord gliomas is a big challenge because of limited options, high recurrence rate and poor prognosis. An intramedullary glioma model is prerequisite for testing new treatments. This paper describes the establishment of a rodent intramedullary glioma model and presents functional progression, neuroimaging and histopathological characterization of the tumour model.</p><p><b>METHODS</b>Fischer344 rats (n = 24) were randomized into two groups. Group 1 (n = 16) received a 5 µl intramedullary implantation of 9L gliosarcomal (10⁵) cells. Group 2 (n = 8) received a 5 µl intramedullary injection of Dulbecco's modified Eagle medium. The rats were anesthetized, the spinous process of the T₁₀ vertebra and the ligamentum flavum were removed to expose the T₁₀₋₁₁ intervertebral space and an intramedullary injection was conducted into the spinal cord. The rats were evaluated preoperatively and daily postoperatively for neurological deficits using the Basso, Beattie and Bresnahan scale. High resolution magnetic resonance images were acquired preoperatively and weekly postoperatively. When score equal to 0, rats were sacrificed for histopathological examination.</p><p><b>RESULTS</b>Rats implanted with 9L gliosarcoma cells had a statistically significant median onset of hind limb paraplegia at (16.0 ± 0.4) days, compared with rats in the control group in which neurological deficits were absent. Imaging and pathological cross sections confirmed intramedullary 9L gliosarcoma invading the spinal cord. Rats in the control group showed no significant functional, radiological or histopathological findings of tumour.</p><p><b>CONCLUSIONS</b>Rats implanted with 9L cells regularly develop paraplegia in a reliable and reproducible manner. The progression of neurological deficits, neuroimaging and histopathological characteristics of intramedullary spinal cord gliomas in rats is comparable with the behaviour of infiltrative intramedullary spinal cord gliomas in patients.</p>

Cocultivation of H2O2-injured astrocytes and bone marrow stromal cells / 中华神经医学杂志

Objective To establish the coculture model of H2O2-injuryed astrocytes and bone marrow stromal cells (BMSCs), observe the effect of BMSCs on self-repair and survival ability of H2O2-injured astrocytes and investigate the mechanism by which BMSCs repair the brain injury.Methods H2O2-injured passage 2 astrocytes were cocultured with passage 3 BMSCs in serum-free medium as coculture group. These two kinds of cells were cultured respectively with the same cell density in same serum-free medium as astrocyte alone group and BMSCs alone group. After 3 d and 7 d cultivation, the morphological changes were observed under light microscope. After 1 d and 3 d cultivation, the supematants from the cell cultures were harvested and enzyme-linked immunosorbent assay (ELISA) method was used to quantify the levels of nerve growth factor(NGF), brain-derived neurotrophic factor(BDNF),basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)in culture supernatants. Results After 3 d and 7 d cultivation, the astrocytes and BMSCs in coculture group grew better compared with astrocyte alone group and BMSCs alone group. The NGF,BDNF and bFGF concentrations in coculture medium were higher than the total of the concentrations in astrocyte alone medium and BMSCs alone medium after 1 d and 3 d cultivation, but the EGF concentration in coculture medium was lower than the total of the concentrations in astrocytes alone medium and BMSCs alone medium after 1 d and 3 d cultivation. Conclusions Coculture of H2O2-injured astrocytes with BMSCs can promote the self-repair ability of astrocytes and increase astrocyte survival rate.Coculture of H2O2-injured astrocytes with BMSCs can also raise NGF, BDNF and bFGF production.The therapeutic efrect of BMSCs for injured brain afcer transplantation might be in part due to the BMSCs conferred protection to astrocytes and increased neurotrophic factor production.

Laboratory living animal model for pure neuroendoscope training / 中华神经医学杂志

Objective To establish a kind of animal model and methods for neuroendoscope training. Methods With rat abdominal cavity hypothesized as cerebral ventricle, a set of programs for neuroendoscopic operative skill training were designed, including endoscopic techiniques, electronic coagulation, suction, flush, biopsy and balloon dilatation, and so on. Results Simulation operation was performed on rat abdomen under neuroendoscope. The procedure helped to practice the endoscopic manipulation, get to know well how to perform endoscope, coagulation, suction, flash instruments, be familiar with the usage of electric coagulation in the liquid condition. The emphasis was put on the basic skills of pure neuroendoscopic operation such as balloon dilatation, electric coagulation, cutting off,forcep biopsy and flush. Conclusions The rat model can provide a way to train pure neuroendoscopic operation. The trainer can get basic experience with regard to endoscopic manipulation, balloon dilatation, electric coagulation, suction, flush and biopsy under endoscope. The method can be an important part of neuroendoscopic laboratory training.

Treatment of pituitary adenomas by simple neuroendoscopic surgery via endonasal transsphenoidal approach / 中华神经医学杂志

Objective To summarize our experience of simple neuroendoscopic endonasal transsphenoidal surgery for pituitary adenoma and discuss its advantages. Methods Forty-two patients with pituitary adenoma diagnosed by CT or MRI detection (diameters of the tumor: <1cm in 11 patients, 1-2cm in 11, 2-3cm in 18, >3cm in 8) were treated by simple neuroendoscopic endonasal transsphenoidal surgery. During transsphenoidal approach, the nasal mucosa was not incised and the nasal septum and median nasal conchae were not dissected. Under neuroendoscopic assistance, the anterior wall of the sphenoidal sinus and sellae base was opened directly and adenoma resection was performed. Results All the patients were followed up for 1-12 months. By postoperative MRI detection, adenomas were resected completely in 38 patients, subtotally in 4 cases. Rapid improvement of vision was noted postoperatively. The increased hormonal level in 18 of 22 patients was decreased to normal postoperatively and another 4 cases improved clinically in serum hormone levels during follow-up. Postoperative transient CSF leakage occurred in 6 cases and transient diabetes insipidus in 23 cases. Conclusions The simple neuroendoscopic endonasal transsphenoidal surgery for pituitary adanoma is safe, time-saving and without severe complications, provides a satisfactory deep illumination and a panoramic operative field. The injury to the patient and postoperative complications are less. The clinical application of the neuroendoscopy for pituitary adenoma is worthy of recommending.

Antitumor activity of F90, an epidermal growth factor receptor tyrosine kinase inhibitor, on glioblastoma cell line SHG-44 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Over-expression of epidermal growth factor receptor (EGFR) is thought to be related to cell proliferation, invasion, metastasis, resistance to chemoradiotherapy and poor prognosis of various human cancers. Forty percent to fifty percent of glioblastoma multiforme (GBM) possess deregulated EGFR, which may contribute to the aggressive and refractory course of GBM. Therefore, blockade of EGFR signal transduction may be a promising treatment strategy for GBM.</p><p><b>METHODS</b>MTT assay, cell growth curve assay and tumor xenograft model were used to evaluate the antitumor activity of F90 against SHG-44 in vitro and in vivo. Western blot assay was applied to evaluate the expression of p-EGFR, p-ERK1, p-JNK, p-P38, Bcl2 and P53 proteins.</p><p><b>RESULTS</b>F90 inhibited the cell proliferation in a dose-dependent manner in vitro. The growth of SHG-44 tumor xenografts was suppressed by F90 at a high dose level (100 mg x kg(-1) x d(-1)). Phosphorylation of EGFR and activated downstream signaling proteins, such as ERK1, JNK and P38, were found to be depressed after incubation with F90 for 48 hours in vitro. Down-regulated Bcl2 protein and up-regulated P53 protein were also observed.</p><p><b>CONCLUSIONS</b>The results demonstrate that F90 is effective in inhibiting the proliferation of SHG-44 cells in vitro and tumor growth in vivo, suggesting that F90 may be a new therapeutic option for treatment of GBM.</p>

Microsurgical and neuro endoscopic anatomy for frontolateral keyhole approach / 中华显微外科杂志

Objective To explore and compare the relevant regional anatomies as they relate the fron- tolateral keyhole approach under microscopy and neuroendoscopy for operations in anterior cranial base and sellar region.Methods Fifteen silieone-injected cadaveric heads were dissected to reveal and compare the extent of expesure through the transfrontolateral keyhole approach under neuroendoscopy and microscopy. Results Portions in the areas of olfactory groove,sellar region and sylvian tissure were blind under micro- scope.Endoscope could allow observation of areas considered blind under the microscope.It could increase light intensity during the approach to objects,extend viewing angles,clear depiction of details in close-up po- sitions and inspect hidden structures.But images of endoscope were two dimensional,lack of view depth.Mi- croscopy and neuroendoscopy could help each other to recuperate deficiency.Conclusion Endoscope-assis- ted neuromicrosurgery is helpful,safe and minimally invasive to treat deepseated lesions in anterior cranial base,sellar region by transfrontolateral keyhole approach.

Reversion of multidrug resistance (MDR) in human glioma cells by RNA interference (RNAi) / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore whether the constructed vector of short haprin in vivo can induce human glioma cell line BT325 to produce RNAi duplexes and reverse the expression of MDR1 gene.</p><p><b>METHODS</b>Three 62nt oligonucleotide fragments (shRNA) were constructed according to GenBank MDR1 sequence and were cloned to the retrovirus-delivered vectors. After transfected these vectors directly into the human malignant glioma BT325 cells by lipofectamine 2000 with enhanced green fluorescence protein (EGFP) co-transfecting, the MDR1 gene silence effects were detected by the changing level of mRNA and P-glycoprotein including real time PCR (RT-PCR), Northern blot and Western blot analysis. To assess the multidrug resistance against adriamycin (ADR) and VCR, cell proliferation assays were performed by cell counting kit-8.</p><p><b>RESULTS</b>The RNAi plasmid vectors were constructed successfully. RT-PCR showed MDR1 mRNA was significantly reduced (P < 0.05). Northern blot analysis showed that the gene silence became most intense at 48 hours after transfection. Western blot analysis demonstrated that P-gp expression was reduced at different time to 12.9%, 30.3% and 4.8%, respectively. The chemosensitivity assays indicated that the transfected cells showed an enhanced sensitivity to ADR and VCR. Based on the value of IC(50), BT325 cells had significantly increased sensitivity to the drugs.</p><p><b>CONCLUSION</b>The sequence specific RNAi can inhibit MDR1 mRNA and P-gp expression in the glioma cell line. It may reverse multidrug resistance phenotype, therefore, may provide promising therapeutic modalities in the treatment of human glioma.</p>

Effects of bone marrow stromal cells on endothelial cells proliferation and microvessel formation in vitro / 中国康复理论与实践

@#ObjectiveTo observe the effects of bone marrow stromal cells (BMSCs) on vessel endothelial cells proliferation and microvessel formation in vitro.MethodsBMSCs and brain vessel endothelial cells were separated from adult and divided into co-culture group of BMSCs and endothelial cells, medium group of BMSCs, comparison group. Endothelial cells proliferation and microvessel formation were observed. ResultsEndothelial cells were promoted to proliferate and formate the microvessel in medium group and co-culture group. And the effect was prominence in co-culture group.ConclusionBMSCs can promote the proliferation and microvessel formation of endothelial cells.

Clinical application of minimally invasive neuroendoscopic techniques / 中国医学科学院学报

<p><b>OBJECTIVE</b>To summarize and analyze the application of neuroendoscopic techniques in neurosurgery, and to discuss the role and significance of neuroendoscopic techniques in the diagnosis and treatment of neurosurgical diseases.</p><p><b>METHODS</b>We treated 1300 patients with different neurosurgical diseases by performing endoscopic neurosurgery (EN) and endoscopy-assisted microneurosurgery (EAM). Among 1300 paitents, 522 were treated with pure endoscopic neurosurgery, 260 with endoscopy-assisted microneurosurgery, 79 with endoscope-controlled bur hole trephination neurosurgery, 434 with endoscope transsphenoidial surgery, and 5 with other techniques through which an endoscope was used in conjunction with stereotactic guidance.</p><p><b>RESULTS</b>Totally 362 hydrocephalus patients were treated using EN. Among them, 190 were treated by third ventriculostomy, 30 by V-P shunt, and 142 patients with complicated hydrocephalus and unsymmetry hydrocephalus by endoscopy-controlled pathologic septum fenestration, septum pellucidum fenestration, and treatment of inventricula inflammation. Clinical symptomatic improvement was achieved in 341 of 362 patients (94.2%). Also 160 intracranial cyst patients were treated using EN for resection and partial resection. Eighty-two patients were performed through cyst-ventricula fenestration. Clinical symptomatic improvement was achieved in 76 of 82 patients (92.7%). Seventy patients treated with endoscopy-controlled bur hole neurosurgery and 8 cases with endoscopy-assisted microneurosurgery got better recovery after operation. Among 260 patients with brain tumors, 252 patients were operated with EAM (190 patients with epidermoid cyst), 8 patients with EN (all brain tumors with diameters < 2.5 cm in inventricular). Clinical symptomatic improvement was achieved in 228 of 260 patients (87.7%). Among 49 patients with inventricular and cistern cyst, 40 patients who were treated by EN and 9 patients by endoscopy-controlled bur hole neurosurgery were resected and their clinical symptoms were improved after operation. Among 434 patients with sellar region lesions, 387 patients with pituitary adenomas, 19 patients with repair for CSF leaks, 9 patients with chordoma, and 19 patients with other neurosurgical diseases were performed with endoscopy-controlled transsphenoidial surgery. Clinical symptoms in 88.9% (386/434) of these patients were improved. Another 5 patients were treated with endoscopy combined with navigation and stereotatic guidance with good results. The complications related to operation were found in only 2% of all the patients including hemorrhage, infection, and damage of important structure.</p><p><b>CONCLUSIONS</b>Clinical application of neuroendoscopic techniques can decrease the damage caused by pure open surgery operation. It is possible to resect lesions at the utmost and protect normal tissue from lesions for using EN and EAM or endoscopy-controlled microneurosurgery (ECM). It is also helpful to enhance surgical quality and, reduce the complications.</p>

Schwann cells transplantation promoted and the repair of brain stem injury in rats / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To explore the possibility of Schwann cells transplantation to promote the repair of injured brain stem reticular structure in rats.</p><p><b>METHODS</b>Schwann cells originated from sciatic nerves of 1 to 2-day-old rats were expanded and labelled by BrdU in vitro, transplanted into rat brain stem reticular structure that was pre-injured by electric needle stimulus. Immunohistochemistry and myelin-staining were used to investigate the expression of BrdU, GAP-43 and new myelination respectively.</p><p><b>RESULTS</b>BrdU positive cells could be identified for up to 8 months and their number increased by about 23%, which mainly migrated toward injured ipsilateral cortex. The GAP-43 expression reached its peak in 1 month after transplantation and was significantly higher than that in the control group. New myelination could be seen in destructed brain stem areas.</p><p><b>CONCLUSION</b>The transplantation of Schwann cells can promote the restoration of injured brain stem reticular structure.</p>