Quality control analysis of brain tissue samples from Shanghai Brain Bank / 解剖学报

Objective To evaluate the feasibility of using brain tissues from Shanghai Brain Bank for the applications on biological research through the analysis of pH value of cerebrospinal fluids, RNA integrity number (RIN), transcriptome, proteome and morphology of brain tissues. Methods The pH value of fresh cerebrospinal fluid was detected by pH test paper; the RNA integrity of cryopreserved brain tissues was examined by Agilent RNA 6000 Nano chip and Agilent 2100 bioanalyzer; the transcriptome sequencing of superior temporal gyrus or caudate was performed using BGIseq-500 sequencer; the proteome of cryopreserved brain tissues was analyzed by Q Exactive HF mass spectrometer; at last, the morphology of the superior temporal gyrus was observed by HE staining. Results The pH value of the cerebrospinal fluid on average was about 6.5. The RIN values of more than 65% of brain tissues were more than 6, indicating good RNA quality. The clean reads ratio after transcriptome sequencing filtering was basically above 80%, indicating that the quality of sequencing library was high. The mass spectrometry analysis of frozen brain samples yielded more than 4000 protein groups and 30 000 peptides, indicating high quality of proteomic data. The morphology of brain tissues was relatively normal, with clearly visible neurons. Conclusion The quality of RNA and protein of brain tissues from Shanghai Brain Bank meets the basic needs for molecular and biological research, and the fixed brain samples can be used for morphological observations.

DNA barcoding research and its application on medicinal plants of Bletilla H. G. Reichenbach / 药学学报

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.

Neuronavigation surgery in China: reality and prospects / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To review the history, development, and reality of neuronavigation surgery in China and to discuss the future of neuronavigation surgery.</p><p><b>DATA SOURCES</b>PubMed, the China Knowledge Resource Integrated Database, and the VIP Database for Chinese Technical Periodicals were searched for papers published from 1995 to the present with the key words "neuronavigation," functional navigation," "image-guided," and "stereotaxy." Articles were reviewed for additional citations, and some information was gathered from Web searches.</p><p><b>STUDY SELECTION</b>Articles related to neuronavigation surgery in China were selected, with special attention to application to brain tumors.</p><p><b>RESULTS</b>Since the introduction of neurosurgical navigation to China in 1997, this core technique in minimally invasive neurosurgery has seen rapid development. This development has ranged from brain structural localization to functional brain mapping, from static digital models of the brain to dynamic brain-shift compensation models, and from preoperative image-guided surgery to intraoperative real-time image-guided surgery, and from application of imported equipment and technology to use of equipment and technology that possess Chinese independent intellectual property rights.</p><p><b>CONCLUSIONS</b>The development and application of neuronavigation techniques have made neurological surgeries in China more safe, precise and effective, and less invasive, and promoted the quality of Chinese neurosurgical practice to the rank of the most advance and excellence in the world.</p>

Clinical experience of 3T intraoperative magnetic resonance imaging integrated neurosurgical suite in Shanghai Huashan Hospital / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Intraoperative magnetic resonance imaging (iMRI) dates back to the 1990s and has been successfully applied in neurosurgery but they were low-field iMRI (< 1.0T). This paper reports the clinical experience with a 3T iMRI-integrated neurosurgical suite in Huashan Hospital, Shanghai, China.</p><p><b>METHODS</b>From September 2010 through March 2012, 373 consecutive patients underwent neurological surgery under guidance with 3T iMRI. A retrospective analysis was conducted regarding clinical efficiency.</p><p><b>RESULTS</b>All surgery in the 373 patients was safe. The ratio of gross total resection for cerebral gliomas (n = 161) was increased from 55.90% to 87.58%. The ratio of benefit in extent of resection was 39.13%. One hundred and fifty eight of the 161 glioma patients accomplished follow-up at 3 months postoperatively. Twenty of 161 patients (12.42%) suffered from early motor deficit after surgery. Late motor deficit was however observed in five of 158 patients (3.16%). Twenty-one of 161 patients (13.04%) had early speech deficit and late speech deficit was only observed in six of 158 patients (3.80%). The ratio of gross total resection for pituitary adenomas (n = 49) was increased from 77.55% to 85.71%. The ratio of benefit in extent of resection was 10.2%. There were no iMRI-related adverse events even for patients who underwent awake craniotomy.</p><p><b>CONCLUSION</b>The 3T iMRI integrated neurosurgical suite provides high-quality intraoperative structural and functional imaging for real-time tumor resection control and accurate functional preservation, resulting in an improvement in maximal safe brain surgery.</p>

Current status of cerebral glioma surgery in China / 中华医学杂志(英文版)

The treatment of gliomas is highly individualized. Surgery for gliomas is essentially for histological diagnosis, to alleviate mass effect, and most importantly, to favor longer survival expectancy. During the past two decades, many surgical techniques and adjuvants have been applied to glioma surgery in China, which lead to a rapid development in the field of cerebral glioma surgery. This article broadly and critically reviewed the existing studies on cerebral glioma surgery and to portrait the current status of glioma surgery in China. A literature search was conducted covering major innovative surgical techniques and adjuvants for glioma surgery in China. The following databases were searched: the Pubmed (January 1995 to date); China Knowledge Resource Integrated Database (January 1995 to date) and VIP Database for Chinese Technical Periodicals (January 1995 to date). A selection criterion was established to exclude duplicates and irrelevant studies. The outcome measures were extracted from included studies. A total of 3307 articles were initially searched. After excluded by abstracts and full texts, 69 studies conducted in the mainland of China were included and went through further analysis. The philosophy of surgical strategies for cerebral gliomas in China is undergoing tremendous change. Nowadays Chinese neurosurgeons pay more attention to the postoperative neurofunctional status of the patients. The aim of the glioma surgery is not only the more extensive tumor resection but also the maximal safety of intervention. The well balance of longer overall survival and higher quality of life should be judged with respect to each individual patient.

Awake craniotomy and intraoperative language cortical mapping for eloquent cerebral glioma resection: preliminary clinical practice in 3.0 T intraoperative magnetic resonance imaging integrated surgical suite / 中华外科杂志

<p><b>OBJECTIVES</b>To evaluate preliminary clinical experience for combining awake craniotomy and intraoperative language brain mapping within the integrated 3.0 T intraoperative magnetic resonance imaging (iMRI) suite.</p><p><b>METHODS</b>From December 2010 to April 2011, 11 right hand-dominant patients with left glioma were involved in, or adjacent to, eloquent cortex was carried out awake craniotomies with cortical stimulation within an integrated 3.0 T iMRI suite. Aphasia battery of Chinese was used to test the language function before the operation. During the procedure, after the occipital, temporal, and supraorbital nerves were blocked by the anesthesiologists, the head was fixed with a custom high-field MRI-compatible head holder. The skull and dura was opened as usual and language brain mapping was then performed. Language testing followed a set protocol: counting numbers from 1 to 50, naming objects, reading single words. Resection of the tumor was guided by neuronavigation system and continued until eloquent areas were encountered or the margin of assessment was reached. An interdissection MRI was acquired to evaluate the glioma removal in a movable MRI scanner after minimal draping. Meanwhile, adverse effects caused by electrical stimulation and iMRI were recorded. The follow-up speech tests were assessed on 7th day and 1 month at least after the operation.</p><p><b>RESULTS</b>The combined use of 3.0 T iMRI and awake craniotomy was performed safely in all patients. No adverse effects were reported. The duration of surgery was prolonged by 2 to 4 h. The patients' perception of iMRI during surgery was favorable. First-look MRI studies led to further resection attempts in 6/11 cases as well as a 3/11 increase in the number of gross-total resections. One week after surgery, baseline language function worsened in 4 cases. However, no patients had a persistent language deficit one month after surgery.</p><p><b>CONCLUSIONS</b>Awake craniotomy and direct cortical electrical stimulation can be performed safely and effectively within a 3.0 T iMRI suite. The combination of high-field iMRI and awake craniotomy may facilitate safe removal of eloquent glioma.</p>

Preliminary application of 3.0 T intraoperative magnetic resonance imaging neuronavigation system in China / 中华外科杂志

<p><b>OBJECTIVE</b>To report the preliminary experience in clinical application of 3.0 T intraoperative magnetic resonance imaging (iMRI) neuronavigation system in China.</p><p><b>METHODS</b>From September 2010 to March 2011, a consecutive series of 122 patients with intracranial lesions underwent operations in guidance with 3.0 T iMRI. A retrospective analysis was conducted regarding clinical efficiency.</p><p><b>RESULTS</b>Among 122 procedures, the numbers of intraoperative scanning were 2 - 4 times with an average of 2.6. The qualities of images were excellent. Due to the discovery and further possibility of resection of residual tumors, the ratio of gross total resection was increased from 71.7% to 90.0% in cerebral gliomas (n = 60), while from 75.9% to 93.1% in macroadenomas (n = 29). There were 6.7% of all patients occurred postoperative paralysis, but only 3.3% of patients had persistent paralysis at 1 - 2 months follow-up. There was no iMRI-related adverse event occurred. During the same period, more than 2500 patients underwent diagnostic MRI scanning.</p><p><b>CONCLUSIONS</b>3.0 T iMRI neuronavigation system provides high-quality intraoperative structural, functional and metabolic images for real time tumor resection control and accurate functional preservation, resulting in an improvement in maximal safe brain surgery. The system is cost-effective.</p>

Thoracoscopic management for bronchiectasis with non-tuberculous mycobacterial infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Non-tuberculous mycobacteria (NTM) have emerged as important opportunistic pathogens of the human being in recent years. Patients with pre-existing bronchiectasis are susceptible to NTM. However, information about its occurrence among bronchiectatic patients in Shenzhen, China is lacking and its impact on the course of bronchiectasis following surgical intervention is unknown. This preliminary study aimed to investigate the prevalence of NTM in bronchiectasis that required surgery in our center, evaluate the role of intraoperative routine screening for NTM, and summarize our initial experience in thoracoscopic management for bronchiectatic patients with NTM.</p><p><b>METHODS</b>A retrospective analysis of clinical, microbiological data of our bronchiectatic patients with NTM over 5 years was made and 40 patients with bronchiectasis were studied to determine the role of intraoperative routine screening for NTM.</p><p><b>RESULTS</b>The prevalence of NTM in this population of patients with bronchiectasis in our center was 6.7% (7/105). The diagnostic yield of the 40 intraoperative specimens was 7.5% (3/40). Of the 7 patients with bronchiectasis and NTM, 3 patients developed postoperative wound infections. All were cured with chemotherapy for 8 - 12 months along with vigorous surgical debridement. Another patient had a slow growth of mycobacteria involving double lungs and the right thoracic cavity and recovered after chemotherapy for nearly 14 months and tube drainage. The affected tissue was completely resected in the remaining 3 patients with no operative mortality and postoperative morbidity, and routine intraoperative screening for NTM was initiated in these patients.</p><p><b>CONCLUSIONS</b>NTM is not uncommon in bronchiectatic patients which deserves surgeons' utmost attention. Routine intraoperative screening for NTM identified otherwise unsuspected patients has shown favorable outcomes. Thoracoscopic management for bronchiectasis with NTM is technically feasible although its role remains to be defined.</p>

Clinicopathologic study of subependymal giant cell astrocytoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features of subependymal giant cell astrocytoma.</p><p><b>METHODS</b>The clinical and pathologic characteristics of 18 cases of subependymal giant cell astrocytoma were retrospectively analyzed.</p><p><b>RESULTS</b>Amongst the 18 cases studied, there was a male predominance (male-to-female ratio = 2:1). The age of patients ranged from 7 to 54 years (mean age = 18.2 years). The tumor often occurred in the lateral ventricles (16/18, 88.9%). Most patients presented with headache and vomiting (11/18, 61.1%), followed by visual disturbance (3/18, 16.7%). Eleven patients (61.1%) had clinical features of tuberous sclerosis, usually in the form of facial angiofibroma (8/18, 44.4%). Computerized tomography was performed in 10 cases, in which 7 cases were of high density and 5 cases showed contrast enhancement. MRI revealed isointense mass lesion on T1WI (7/11, 63.6%), highly intense lesion on T2WI (10/11, 90.9%) and contrast enhancement in some cases (9/11, 81.8%). Four patients had follow-up information available and all of them were alive from 1 to 5 years (mean = 3.5 years). Histologically, there were bundles of spindle cells mixed with clusters of gemistocytes and ganglion-like cells. The spindle cells showed immunoreactivity for glial fibrillary acidic protein (18/18, 100%), while the gemistocytes and ganglion-like cells expressed synaptophysin (14/18, 77.8%). Most of the cases (16/18, 88.9%) had MIB-1 index <or= 1%.</p><p><b>CONCLUSIONS</b>Subependymal giant cell astrocytoma is a benign brain tumor with distinctive histopathologic features. The tumor typically affects children and young adults. It is associated with a favorable clinical outcome, especially if completely excised.</p>

Application of virtual reality technique in preoperative planning of neurosurgery / 中华显微外科杂志

Objective To evaluate the usefulness of virtual reality technique in preoperative planning of neurosurgery.Methods Multiple medical imaging data including magnetic resonance imaging,magntic rsonanc angiograph,magnetic resonance venography and computed tomography collected from 26 patients who suffered craniocerebral diseases,were transfered to Dextroscope system.A stereoscopic 3-D object displayed after merging,extracting,dissecting the data with the Radiodexter software in virtual reality environment.A suite of 3-D tools accessible inside the Dextroscope workspace enabled users to obtain measurement and simu- lated intraoperative viewpoint about the lesion and adjacent anatomic structure.A preoperative plan was de- fined and compared with real operation.Results 3-D stereoscopic virtual reality images of 26 cases were re- constructed successfully,and of coincidence with real situation in operations.Conclusion VR technique in Dextroscope system can integrat multiple medical imaging data quickly,intuitively and overall,offer a compre- hensive information about the lesions and related local anatomy,give a hand to optimizing the operative project and might be possible potentially to increase the safety of operation and the resecting rate of the lesion.

Prospective comparison of functional magnetic resonance imaging and intraoperative motor evoked potential monitoring for cortical mapping of primary motor areas / 中华外科杂志

<p><b>OBJECTIVE</b>To compare the relation between the preoperative functional magnetic resonance imaging (fMRI) with blood oxygen level dependent (BOLD) technique and intraoperative motor evoked potential (MEP) monitoring for cortical mapping of primary motor cortex in patients with tumors near the central area. And to determine whether non-invasive preoperative fMRI can provide results equivalent to those achieved with the invasive neurosurgical "gold standard".</p><p><b>METHODS</b>A prospective study of 16 patients with various pathological tumors of the central area was conducted. Preoperative fMRI scans using the BOLD contrast technique in each patient were performed. An activation scan was achieved by using a motor task paradigm, which consisted of simple flexion-extension finger movements and finger-to-thumb touching in a repeating pattern. The anatomical structure was delineated by the T(1)-weighted three-dimensional fast spoiled gradient recalled sequence (3D/FSPGR) immediately afterward. The BOLD images were overlaid on the T(1)-weighted 3D/FSPGR images, and then co-registered to the neuronavigation system. The fMRI activations were documented by using a neuronavigation system in sequence, and compared to standardized intraoperative MEP monitoring, which included direct cortical electrical stimulation (DCES) or transcranial cortical electrical stimulation (TCES) or their combination. The compound muscle action potentials of forearm flexor and hand muscle responses were recorded during either TCES or DCES. Two techniques were compared to determine the accuracy for cortical mapping of primary motor areas with fMRI.</p><p><b>RESULTS</b>Overall, the intraoperative MEP monitoring showed good correlation with fMRI activation in 92.3% of cases. The coincidence rate, however, was 100.0% between TCES and fMRI, and 66.7% between DCES and fMRI respectively. There was no statistically difference between two cortical mapping techniques, chi-square test of paired comparison of enumeration data, P < 0.01.</p><p><b>CONCLUSION</b>BOLD fMRI was a high sensitive and reliable technique to locate the position of the primary motor areas and their spatial relation with adjacent tumor, especially for the presurgical planning in patients with central area brain tumor.</p>

Role of diffusion tensor imaging in neuronavigation surgery of brain tumors involving pyramidal tracts / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the role of diffusion tensor imaging (DTI) in neuronavigation surgery of brain tumors involving pyramidal tracts.</p><p><b>METHODS</b>Forty-nine patients with brain tumors involving pyramidal tracts were randomly divided into trial group (DTI navigation) and control group (traditional navigation). The patients in trial group underwent DTI and T1 weighted 3D navigational magnetic resonance imaging (MRI) studies. The main white matter tracts were constructed by the DTI datasets, and merged to the anatomical structure, which was delineated by the T1-weighted three-dimensional fast spoiled gradient recalled sequence (3D/FSPGR). The relationship between the tumors and adjacent pyramidal tracts were segmented and reconstructed for three-dimensional visualization.</p><p><b>RESULTS</b>In 25 patients of trial group and 24 patients of control group, the statistic analysis confirmed well balance of main variations. The tumors were completely resected in 12 patients (50.0%) of control group and in 20 patients (80.0%) of trial group (P < 0.05). Postoperative aggravated contralateral extremities weakness or hemiplegia due to pyramidal tract injury occurring in 75.0% cases of control group whereas only 20.0% patients in trial group (P < 0.01). The mean Karnofsky scale were 69.58 +/- 23.49 and 84.80 +/- 23.49 respectively in control and trial groups (P < 0.05). The excellent outcome ratio (Karnofsky scale = 90 - 100) was 37.5% in control group and 72.0% in trial group respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>DTI allows individual estimation of large fiber tracts of brain. Furthermore, to integrate spatial three-dimensional information concerning the white matter tracts into traditional neuronavigation images during surgery, was valuable in presenting topographical character of involving (shift or erosive) pyramidal tracts and relationship with the margins of neighboring tumors. The mapping of large fiber tracts was a safe, efficient, reliable technique. DTI should be routinely used in neuronavigation surgery of brain tumor involving pyramidal tracts to plan the optimal trajectory and ensure total resection of the lesions during operation, as well as to decrease potential disability after operation and to shorten the length of hospitalization.</p>

Construction of combined site-directed random mutation libraries of recombinant human lymphotoxin / 生物工程学报

To construct the combined site-directed random mutation library of recombinant human Lymphotoxin (rhLT) for in vitro molecular evolution study, and to study the structure and function relationship. The random point mutations at the sites of 46,106 and 130 were individually generated by overlap PCR amplification with the random nucleotide primers. The three point mutations were combined and cloned into pMD-18T vector to construct the combined mutation library. DNA sequencing was used to evaluate the diversity and randomness of the mutation sites. The combined mutation library was re-engineered, inserted in prokaryotic expression vector pBV220, transformed and expressed in Escherichia coli strain DH5alpha. The biological activity of some of the mutants was tested in 1929 mouse fibroblast cells. As much as 1.5 x 10(5) clones were obtained, which represents 4.5 times of the complete mutation libraries at 99% confidence. Sequencing 50 clones revealed no obvious bias in the nucleotide and amino acid mutations at the sites. Among the 30 expressed samples underwent for the bioassay, 70% (21 samples) were inactive, 23.3% (7 samples) had lower activity than rhLT, the remaining 6.7% (2 samples) had higher activity than rhLT. The combined site-directed random mutation library of rhLT has been constructed successfully. In combination with phase display, the library is ready for in vitro molecular evolution study.

The application of indirect immuno-fluorescence assay in the diagnosis of severe acute respiratory syndrome / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the temporal profile of serum antibody against coronavirus in patients with severe acute respiratory syndrome (SARS), and to evaluate the reliability of indirect immuno-fluorescence assay (IFA) in the diagnosis of SARS.</p><p><b>METHODS</b>Clinically confirmed SARS patients, suspected SARS patients, and controls were included in the study. IFA was used to detect the serum antibody against SARS coronavirus. General information about the subjects was collected using a standard questionnaire.</p><p><b>RESULTS</b>The positive rates of specific IgG and IgM against SARS virus within 10 days after onset of the disease were 55.1% and 16.3% respectively and then increased up to 89.8% for IgG and 65.3% for IgM. After 25 days of the onset of the disease, 90.9% patients became positive for both IgG and IgM. Results from chi-square for trend test revealed that the positive rates of both IgG and IgM increased with time (chi(2) for trend = 16.376, P = 0.00005 for IgG; chi(2) for trend = 28.736, P = 0.00000 for IgM). Sensitivity, specificity and agreement value of IFA regarding the diagnosis of SARS were all higher than 90%.</p><p><b>CONCLUSION</b>IFA can be used to assist diagnosis of SARS after 10 days of the onset of disease.</p>