BACKGROUND: The neural stem cells (NSCs) will be clinically applied extensively in the future, and seeking of more promoting methods of the proliferation and differentiation of NSCs in vitro will be the study direction of NSCs.OBJECTIVE: To introduce an economic and time-saving method for NSCs proliferation.DESIGN: Observation and control experiment.SETTING: Beijing Neurosurgical Institute.MATERIALS: Four Wistar rats pregnant for 14 days with the body mass of (180±20) g (bought from Animal Department of Chinese Academyof Medical Sciences with the batch number of SCXK1100-0006 for experiment animals) were selected and fed at common temperature and humidity.The DMEM/F12 and B27 were bought from Gibco Corporation. The basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were bought from PeproTech Company. The nidogen monoclonal antibody (MA),glial fibrillary acidic protein (GFAP) polyclonal antibody, galactocerebroside polyclonal antibody and microtubule-associated protein 2 (MAP2) were obtained from Chemicon Company. The fetal bovine serum (FCS) was provided by Hyclone company.METHODS: The experiment was conducted in the Department of Injury and Repair of Beijing Neurosurgical Institute from May to October 2004.Wistar rats were executed by dislocation and sterilized by putting into 75% alcohol. Four rats were used each time, the brain tissues of which were put into Hank's fluid. The cerebral pia mater and vessels were isolated under anatomical microscope, and the brain tissues were sheared off with eye scissors, which were filtered by 200 mesh copper grid and collected into 2 centrifuge tubes, and the supernatant was gotten rid off after that. ①Cells were divided into serum pre-culture group and control group according to whether there were serum pre-culture. The DMEM nutrient fluid of FCS (100 g/L) was added to serum pre-culture group, which was replaced by DMEM/F12 nutrient fluid of EGF, bFGF and B27 at 48 hours after culture. The cells in the control group were added with DMEM/F12 nutrient fluid of EGF, bFGF and B27 to culture in 5% CO2 incubator for one week at 37 ℃. The growth of NSCs at 48 hours after culture was observed in both groups under inverted contrast phase microscope.②On the 5th and 10th day after differentiation induced by 100 g/L FCS, nestin, GFAP, galactocerebroside and MAP2 were stained with immunofluorescence antibody,and the expressions of NSCs in both groups were observed under fluorescent microscope. The PBS buffer solution was used instead of first antibody in the control group, while other procedures were the same as above.MAIN OUTCOME MEASURES: ①Observation of NSCs growth at 48 hours after culture under contrast phase microscope in both groups. ②Detection of NSC specific antigen expressions with immunofluorescence stain in both groups.RESULTS: ①In the cells of the serum pre-culture group at 48 hours after culture, there were large number of regular NSC bulbs, most of which were aggregated by 8-10 cells. The proliferation of cell bulbs was accelerated after the nutrient fluid was replaced to DMEM/F12 nutrient fluid of EGF,bFGF and B27, while irregular lamellar cells could be seen in the control group at 48 hours after culture, and small regular cell bulbs were found at 4-5 days later. ②It could be seen under fluorescence microscope that on the 5th day after induced differentiation, the nestin, GFAP and galactocerebroside were positive, while MAP2 was negative. On the 10th day after induced differentiation, nestin and GFAP were positive, whereas the galactocerebroside and MAP2 were negative (no representation).CONCLUSION: Serum pre-culture can accelerate the bulb-aggregation of NSCs as well as promote the proliferation of NSCs.

For decades, the selection of chemotherapeutic regimens for the treatment of recurrent or metastatic nasopharyn-geal carcinoma has been mainly empirical. To our knowledge, there is no phase 3 trial that has been conducted to determine the optimal treatment for these patients before our publication. Recently, we published an article inThe Lancet entitled “Gemcitabine plus cisplatin versus lfuorouracil plus cisplatin in recurrent or metastatic nasopharyngeal carcinoma: a multicentre, randomised, open-label, phase 3 trial.” The results of our study indicate that gemcitabine plus cisplatin could improve the survival of patients with recurrent or metastatic nasopharyngeal carcinoma com-pared with conventional lfuorouracil plus cisplatin.

BACKGROUND:Cerebrovascular drag, occlusion and other mechanical stimulations inevitably occur during some craniocerebral operations, which cause acute mechanical cerebrovascular vasospasm. At present, the mechanism underlying the patho-physiology as well as the pathological prognosis of this acute mechanical vasospasm remains unclear.OBJECTIVE: To observe changes in the vascular diameter of the middle cerebral artery, cerebral blood flow (CBF), and ultrastructure of vascular wall and vascular endothelial cells, during the early stage (2 hours) of mechanical cerebral vasospasm in cats.DESIGN: Open experiment.SETTING: Department of Neurology, Beijing Tiantan Hospital, Capital Medical University; Beijing Institute of Neurosurgery.MATERIALS: Six healthy adult hybrid cats, of either gender, weighing from 2.5 to 3.5 kg, were provided by the China Medical Science Institute of Experimental Animals. Laser Doppler flowmetry (Periflux 5010, Sweden Perimed Company)was used.METHODS: This study was carried out in the Beijing Institute of Neurosurgery between August 2005 and March 2006. For all experimental surgical procedures, the cats were anesthetized by intraperitoneal injection of 200 g/L chloral hydrate, at 2 mL/kg, and then placed in a prone position. A median incision was made in the scalp and a square bone window, 8×10 mm, was opened at 1.5 cm posterior and 1.5 lateral to the anterior fontanel, after which the dura mater was pricked out. The fine detecting head of the Laser Doppler flowmetry was fixed to a region of the cerebral surfacewith no vessels or with only a few vessels. Subsequently, the cats were placed in lateral position. Under the surgical microscope, the right middle cerebral artery was exposed through a suborbital approach. Blunt apparatus was used to stimulate middle the middle cerebral artery repeatedly, at a frequency of 100 time/min within 30 minutes.The diameter of the middle cerebral artery was measured and a perfusion index of cortical brain tissue was monitored, separately, before and then at 0, 0.5, 1.0, 1.5, 2.0 hours after stimulation. Ultrastructural changes in the vascular wall and the vascular endothelial cells were observed during the early stage (2 hours) of mechanical cerebral vasospasm in cats.MAIN OUTCOME MEASURES: Diameter of the middle cerebral artery and the perfusion index of cortical brain tissue before and then at 0, 0.5, 1.0, 1.5, 2.0 hours after stimulation, as well as any ultrastructural changes in the vascular wall and endothelial cells 2 hours after stimulation.RESULTS: The results from six cats were were analyzed. ①The diameter of the middle cerebral artery was (0.617±0.129), (0.723±0.082), (0.840±0.084) mm 0, 0.5 and 1.0 hours after stimulation, respectively, which was significantly smaller than that before stimulation [(0.897±0.066) mm,t =4.74, 4.017, 1.299,P ＜ 0.01]. ② The perfusion index of cortical brain tissue was 67.8±18.5, 82.5±17.5, 89.8±24.0, 94.0±22.2 and 98.5±21.0 at 0, 0.5, 1.0,1.5 and 2.0 hours after stimulation, which was significantly lower than that before stimulation (159.2±23.5, t =4.716-7.469, P ＜ 0.01 ). ③ At the early stage of acute mechanical stimulation (2 hours) to middle cerebral artery, endothelial cell chromatin aggregated at the edge of the cells and achromocyte formed, but mitochondrial crista was unclear.CONCLUSION: Mechanical stimulation to the middle cerebral artery in cats can lead to cerebral vasospasm. Apoptosis of endothelial cells appears at the early stage of stimulation (2 hours). These results indicate that, in order to prevent against cerebral vasospasm, cerebrovascular mechanical stimulation should be as minimal as possible and that as few as possible craniocerebral operations should be performed.

ObjectiveTo observe the changes of artery diameters, cerebral blood flow and glutamate level in the early stage (2 h after the stimulus finished) of acute mechanical middle cerebral artery (MCA) vasospasm in cats. MethodsThe right MCA was persistently mechanically stimulated using a small smooth stainless steel nail in the field across the olfactory tract for 30 min. The diameter of MCA was recorded with metrical ocular of microscope. The changes of the perfusion index of brain tissue were observed through the Laser Doppler flowmetry monitor fixed on the skull. The level of glutamate were investigated through high performance liquid chromatography. ResultsThe diameter of MCA decreased to 68.8% of normal. 2 h later, the diameter of MCA recovered. The perfusion index of the cortex surface decreased to 42.6% of normal and up to 63.8% 2 h later. The level of glutamate raised about 40 times of normal and maintained a high level 2 h after the mechanical stimulus. ConclusionThe persistent mechanical stimulus can cause acute cerebral vasospasm. Reduce of cerebral blood flow and raise of excitatory amino acids were observed in the early stage of acute mechanical vasospasm.

ObjectiveTo introduce a method of promoting neural stem cells proliferation.MethodsNeural stem cells were cultured with medium DMEM containing 10% fatal bovine serum for 48 h, then changed medium DMEM/F12 containing epidermal growth factor, basic fibroblast growth factor and B27. Immunohistochemistry was used to identify nestin expression.ResultsNeural stem cells got together fast and formed neurospheres within 48 h and expressed nestin. ConclusionPro-culture with serum can promotes neural stem cell proliferation.

Objective To explore mRNA and protein expression of PAl-1 in kidney tissue of mice with IgA nephropathy (IgAN) and the effects of YiShen Decoction. Methods The IgAN model was built by the method of oral intake of bovine serum albumin(BSA)together with the injection of staphylococcus enterotoxin B(SEB)through caudal vein. The mRNA expression of PAI-1 was measured by fluorescent quantitative polymerase chain reaction (FQ-PER)and the expression of PAI-1 protein was measured by immunohistochemistry. Results No significant difference of mRNA and protein expression of PAI-1 was found between the low concentration and high concentration Yishen Decoction group. But the secretion and mRNA expression of PAI-1 in the low and hiigh concentration Yishen Decoction group was decreased more than that in the IgAN model group. Conclusion The abnormal expression of PAI-1 mRNA and protein played an important role in the onset and development of IgAN. The TCM Yishen Decoction could reduce the abnormal expression of the mRNA and protein of PAI-1 in kidney tissue of mice with IgA nephropathy.

Background: Bone metastases are common in patients with advanced cancer. Bisphosphonates (BPs) could prevent or delay the development of skeleton-related events (SREs). The present study aimed to identify the clinical features of and treatment strategies for Chinese patients with bone metastases. Methods: Consecutive cancer patients who had bone metastases and received BP treatment were enrolled. A ques-tionnaire was developed to collect the patients’clinical data, as well as information on the diagnosis and manage-ment of bone metastases. Physicians’awareness of the guidelines and knowledge of the application of BP were also assessed. Results: A total of 3223 patients with lung cancer (36.5%), breast cancer (30.9%), prostate cancer (8.5%), and gas-trointestinal cancer (5.7%) were included in this study. The sites of bone metastases were the thoracic spine (56.0 %), lumbar spine (47.1%), ribs (32.6%), and pelvis (23.2%). The SRE frequency was the highest in patients with multiple myeloma (36.6%), followed by those with lung cancer (25.9%), breast cancer (20.2%), prostate cancer (18.2%), and gas-trointestinal cancer (17.3%). Irradiation to the bone was the most frequent SRE (58% in lung cancer patients, 45% in breast cancer patients, and 48% in prostate cancer patients). Our survey also showed that 45.5% of patients received BP within 3 months after their diagnosis of bone metastases, whereas the remaining 54.5% of patients did not receive BP treatment until at least 3 months after their diagnosis of bone metastases. The SRE frequency in the former group was significantly lower than that in the latter group (4.0% vs. 42.3%, P < 0.05). In patients with more than 6 months of continuous BP treatment, the mean time to the first SRE was significantly longer than that in patients with less than 6 months of continuous BP treatment (7.2 vs. 3.4 months, P < 0.05). In addition, 12.2% of the physicians were not aware of the efcacy of BP in preventing and delaying SRE. Only half (52.3%) of the physicians agreed that the BP treatment should persist for at least 6 months unless it was intolerable. Conclusions: Our study suggested that prompt and persistent BP treatment was associated with a reduced risk of SREs. However, our survey also revealed that the proper application of BP was not as common as expected in China.

Objective:To investigate the clinical efficacy of different grade transoral atlantoaxial release for the treatment of irreducible atlantoaxial dislocation.Methods:From January 2010 to December 2019, 297 patients with irreducible atlantoaxial dislocation treated by different grade releases were retrospectively analyzed, including 132 males and 165 females, aged 42.3±12.14 years (range, 10-63 years). All cases were treated by different grade releases, Grade I (196, 66.0%), Grade II (54, 18.2%), Grade III (28, 9.4%) and Grade IV (19, 6.4%). The American Spinal Injury Association (ASIA) grade and Japanese Orthopedic Association (JOA) score were recorded as the clinical evaluation index. The clivus-canal angle (CCA) and cervico-medullary angle (CMA) were measured to evaluate the reduction. The surgery time, blood loss, duration of bony fusion and complications were also analyzed.Results:The follow-up time was 14.8±10.2 months (range, 9-36 months). The surgery time of Grade I-IV were 2.02±0.35 min, 3.00±0.36 min, 4.07±0.96 min and 5.24±0.83 min, respectively ( F=385.43, P<0.001), blood loss was 84.08±27.21 ml, 153.61±31.36 ml, 268.93±48.94 ml and 444.21±109.51 ml, respectively ( F=582.39, P<0.001). The preoperative ASIA motor score of Grade I-IV were 83.85±6.68, 84.06±5.47, 84.07±5.99 and 85.00±4.11, respectively. The last follow-up were 98.34±2.38, 98.67±1.79, 98.86±1.58 and 98.32±2.11, respectively, with statistically significant differences from preoperative ( P<0.05). The preoperative JOA score of Grade I-IV were 11.44±1.73, 11.59±1.72, 11.61±1.47 and 11.32±1.80, respectively. The last follow-up were 16.22±1.00, 16.28±1.02, 16.14±1.04 and 16.16±1.07, respectively, with statistically significant differences from preoperative ( P<0.05). The preoperative CCA of Grade I-IV were 110.19°±8.76°, 112.48°±7.66°, 106.61°±6.54° and 109.05°±7.79°, respectively. The last follow-up were 140.22°±8.04°, 141.86°±7.04°, 142.35°±8.62° and 140.15°±6.49°, respectively, with statistically significant differences from preoperative ( P<0.05). The preoperative CMA of Grade I-IV were 113.48°±9.54°, 116.03°±8.38°, 109.55°±7.13°, and 112.46°±8.33°, respectively. The last follow-up were 144.28°±7.75°, 146.40°±6.98°, 145.81°±8.27° and 143.24°±6.36°, respectively, with statistically significant differences from preoperative ( P<0.05). Solid bony fusion was obtained except for 3 cases, the fusion time was 9.71±2.55 months (range 3-14 months). Altogether 33 complications occurred in all cases (11.1%), including 3 fusion failure, 3 cerebrospinal leak, 3 wound infection, 2 death (1 case caused by cerebrospinal leak), 11 pharyngeal discomfort, 4 postoperative pain surrounding iliac crest, and 8 malunion of iliac crest. Conclusion:Transoral stepped atlantoaxial release theory could provide guidelines for atlantoaxial dislocation treatment, and make the transoral release technique more effective and safer.

Objective To explore the association between the ABO blood group and the risk of myocardial infarction in Chinese people.Methods We retrospectively recruited 1 988 consecutive patients with acute myocardial infarction (AMI) and 1 856 non-coronary artery disease (non-CAD) subjects who hospitalized in our hospital between January 2013 and December 2013.The clinical features and ABO blood group were analyzed.Results Blood group distribution was A (27.1％,539/1 988),B (34.4％,684/1 988),AB(10.8％,215/1 988),O (27.7％,551/1 988) in patients with AMI and A (26.7％,496/1 856),B(32.2％,598/1 856),AB (10.8％,200/1 856),O (30.4％,564/1 856) in non-CAD group.The single factor analysis showed that blood group O tended to be more common in the non-CAD group than in AMI group (P =0.06).After adjustment for common cardiovascular risk factors such as age,gender,hypertension,diabetes,smoking and serum cholesterol level,the A,B,and AB blood groups were associated with increased risk of AMI compared with O blood group,and the difference was significant with A blood group (OR =1.229,95％ CI 1.019-1.482,P =0.031) and B blood groups (OR =1.214,95％ CI 1.017-1.449,P =0.032).In addition,non-O blood group remained significantly associated with the increased risk of AMI than O blood group after logistic regression analysis(OR =1.223,95％ CI 1.048-1.426,P =0.01).Conclusion Our results suggest that non-O blood group is associated with the increased risk of AMI.

ObjectiveTo investigate the levels of neutralizing antibodies against the novel coronavirus in the serum of elderly individuals aged 60 years and above in Shanghai’s Changning District， following natural infection and mixed immunity， in order to provide a basis for strengthening immunity in the elderly. MethodsElderly people who participated in free health check-ups at 10 community health service centers in Changning District from May to June 2023 were selected as the subjects. Information such as gender， age， COVID-19 infection history， COVID-19 vaccine immunization history， and chronic disease history were collected. Serum samples of the subjects were collected and quantitative detection of SARS-CoV-2 neutralizing antibodies was performed by magnetic particle chemiluminescence method. The antibody levels of different populations were analyzed. ResultsA total of 620 subjects were included， 586 of whom （241 males and 345 females） met the study conditions. There were 90 people in the full vaccination + infection group， 224 people in the intensive vaccination + infection group， and 272 people in the unvaccinated + infection group. The positive rates of COVID-19 antibody in the three groups were 94.44% （95%CI： 87.51%‒98.17%）， 95.98% （95%CI：92.51%‒98.15%） and 22.06% （95%CI： 17.28%‒27.46%）， respectively. The positive rates in full vaccination + infection group and intensive vaccination + infection group was significantly higher than that in unvaccinated + infection group （χ²=147.561，P<0.01；χ²=271.729，P<0.01）. The antibody level in full vaccination + infection group （640.74 AU·mL^-1） and intensive vaccination + infection group （1 200.88 AU·mL^-1） was significantly higher than that in unvaccinated + infection group （4.51 AU·mL^-1） （all P<0.01）.The antibody level in the intensive vaccination + infection group was also significantly higher than that in the whole vaccination + infection group （P < 0.05）. ConclusionAfter 5‒6 months of infection， the neutralizing antibody positive rate and antibody level were significantly higher in the elderly who received the full vaccination and infection or intensive vaccination and infection. It is recommended that elderly individuals， who have been infected for more than 5‒6 months but have not been vaccinated， should consider getting vaccinated to enhance their levels of neutralizing antibodies.