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.

Objective: By constructing the cash flow model, it proposed the premium adjustment mechanism of critical illness insurance(CII). Methods: Based on the practical data of critical illness insurance in China, it established some actuarial assumptions and cash flow model to simulate and analyze premium adjustment mechanism of CII. Results: ( 1) Because of the deterioration of critical illness incidence rate and the change of market interest rates , CII guaranteed premium usually resents pricing risk, which showed that the cash flow is negative, sometimes accumulated cash is negative; (2) based on the criteria of insurance cost adjustment, it is suggested that when the loss rate reached more than 70%, which could permit the insurance company adjust the premium, if the loss rate reached more than 80%, it needs to compulsory the insurance companies to adjust premium. Conclusion: The study proposes the mechanism of CII premium adjustment, provides guidance for practice.

Objective To evaluate the value of multi-slice spiral CT (MSCT) angiography in diagnosis of thoracic vascularemergency.Methods 25 cases suspected of thoracic vascular emergency were examined with MSCT. Two dimensional and three dimensionalimaging methods were used. Results Central pulmonary embolism 10 cases, aortic dissection 12 cases,aneurysm 2 cases and takayasuarteritis with stricture of kidney artery were found by two dimensional imaging(MPVR,CPR),and the extent, size and details of lesions couldbe showed well. The space relationship and calcifation of aorta were displayed in three dimensional imaging(SSD,VR,MIP). Therelationship between intraluminal embolism and the wall of pulmonary artery, as well as intimal flap and reentry of dissection could bevealuated by CTVE.Conclusion Associated with imaging post processing,MSCT angiography is an effective method in diagnosis of thoracic vascular emergency.

lso.This result suggested that Th17 subset is differentiated in chronic stage of viral myocarditis.

Objective To study the correlation between high resolution computed tomography manifestations and expression of transforming growth factor beta,tumor necrosis factor alpha in radiationinduced lung injury of rats,and to investigate the vslues of cytokine detection and HRCT scanning for the prediction and early diagnosis of radiation-induced lung injury.Methods Forty-eight Sprague-Dawley (SD)rats were randomly divided into eight groups,group A was normal control group.and group BH were irradiated with a single dose of 15 Gy to the lungs.HRCT scanning Wag performed before and 1 week,2,4,8,12,16,24 weeks after radiation in group A-H respectively.The expression of TGF-beta and TNF-alpha were detected with ELISA. All the rats were killed to observe pathological changes of their lungs.HRCT signs,levels of cytokine were simultaneously compared and analyzed.The t-test and Spenrman rank correlation were used for the statistics.Results Four HRCT signs were observed during the 24 weeks after radiation,including ground-glass opacity(1 case),patchy consolidation(8 cases),massive consolidation(7 cases)and fibrosis(3 cases).The average levels of TGF-beta in group B-H[(3.33±0.47),(3.20±0.65),(3.12±0.45),(3.54±0.80),(3.30±1.13),(2.49±0.67),(4.19±0.22)μg/L,respectively]were hisher than the control group[(0.45±0.14)μg/L.P＜0.05].At 24 weeks the average level increased to the highest peak[(4.19 4-0.22)μg/L,P＜0.05].At 1 week and 2,4,8,12 weeks after radiation,the average level of TNF-alpha in radiation group[(236.52±29.01),(214.91±34.53),(270.97±42.12),(208.83±86.51),(208.83±82.23)ng/L]was hisher than the control group[(31.78±0.92)ng/L,P＜0.05].The average level increased to the highest peak [(270.97±42.12)ng/L,P＜0.05]at 4 weeks.The average level at 16 and 24 weeks[(60.63±38.49),(32.07±1.05)ng/L]dropped to the level similar to the control group(P＞0.05).There were no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha(rs=0.5570 and 0.1013.P＞0.05).HRCT signs were correlated with pathological changes.Conciusions The monitoring of TGF-beta and TNF.alpha in the serum after irradiation can predict the development of radiation-induced lung injury.There are no rank correlations between HRCT manifestations and expression of TGF-beta and TNF-alpha.

Objective To assess the efficacy and adverse reaction of Lanthanum carbonate in patients with hyperphosphatemia and high calcium times phosphate product receiving maintenance hemodialysis.Methods Twenty-three hemodialysis patients with serum phosphate ≥ 2.0 mmol/L and/or the serum calcium times phosphate product ≥ 4.52 mmol2/L2 were prescribed Lanthanum carbonate chewed with meals for 4 weeks.Blood (Hb), serum albumin (ALB), alanine amino shift enzyme (ALT), aspartic acid transaminase (AST) ,calcium(CA), phosphorus (P) calculated the product of calcium and phosphorus (Ca×P) and intact parathyroid hormone (iPTH) level were checked before and after Lantharum carbonate therapy.The adverse reactions were recorded concomitantly.Results Serum phosphate and the serum calcium time phosphate product significantly decreased after 1 month Lanthanum carbonate therapy((2.48±0.55) mmol/L vs.(1.83 ±0.37) mmol/L, (5.21 ± 1.35) mmol2/L2 vs.(4.10±0.96) mmol2/L2;t =2.742,2.936;P＜0.05).Serum calcium, iPTH level and blood cells count remained no change(P＞0.05), while serum ALT, AST concentration within normal range.Five patients complained affordable abdomen discomfort or upset.Conclusion Lantharum carbonate show to effectively and quickly control serum phosphorus levels and calcium time phosphate product in hemodialysis patients without inducing changes in serum calcium and should be considered an ideal phosphate binder.

Objective To analyze the clinical characteristics of Klebsiella pneumoniae infection in preterm infants. Methods Clinical data of 75 preterm infants infected with Klebsiella pneumoniae treated in BaYi Children's Hospital from February 6,2008 to February 10,2010 were retrospectively analyzed.The difference of auxiliary examination between early-onset and late-onset infection group were compared by two independent samples t test.Spearman correlation analysis and non-conditional Logistic regression analysis were used to analyze the high risk factors and the prognostic factors of Klebsiella pneumoniae infection in preterm infants. Results The incidence of Klebsiella pneumoniae infection was 2.8％ (75/2721) in preterm infants,and the mortality rate was 9.3％ (7/75). There were 71 cases of Klebsiella pneumoniae sepsis and 4 cases of Klebsiella pneumoniae pneumonia.Among 75 cases,63 cases were early-onset infection (onset age≤72 h) and 12 were late-onset infection (onset age＞72 h).All patients presented with poor response,heart rate during quiet sleep ＞ 160/min and low oxygen saturation.The mean corpuscular volume and mean corpuscular hemoglobin concentration in early-onset Klebsiella pneunoniae infection cases were higher than those in late-onset neonates [(128.87±24.60) fl vs (113.72±13.54) fl,t=-2.07,P＜0.05and (38.11±2.15) pg vs (36.98±1.05) pg,t=-2.76,P＜0.05].Low birth weight and caesarean section were associated with early-onset Klebsiella pneumoniae sepsis (r=0.250 and -0.240,P＜0.05). The prognosis of Klebsiella pneumoniae infection was associated with hospital stay and duration of premature rupture of membranes (r=0.368 and 0.318,P＜0.05). Conclusions There were no specific clinical manifestations for Klebsiella pneumoniae infection in preterm infants.Preterm infants with low birth weight,long duration of premature rupture of membranes,delivered by caesarean section and received invasive operation are likely to develop Klebsiella pneumoniae infection.

Objective:To analyze the clinical and gene mutation characteristics of congenital disorder of glycosylation (CDG) caused by compound heterozygous mutation of the COG6 gene ( COG6-CDG). Methods:This study retrospectively analyzed the clinical data and genetic test results of a patient with COG6-CDG in Bayi Children's Hospital, the Seventh Affiliated Medical Center of Chinese PLA General Hospital, in August 2019. Literature was retrieved with keywords including COG6, COG6-CDG, congenital disorders of glycosylation typeⅡL and congenital disorders of glycosylationⅡL in China National Knowledge Infrastructure, Wanfang Database, VIP Database, PubMed, and Web of Science Database from the establishment to July 2020, to summarize the clinical and genetic characteristics of COG6-CDG. Results:(1) Case report: The 59-day-old baby boy, with a gestational age of 27 +5 weeks and birth weight of 1 180 g, presented with multi-system involvement on admission, including unidentified progressive hepatosplenomegaly with jaundice and ascites, persistent thrombocytopenia, microcephaly, hypotonia, hypohidrosis, hyperkeratosis, and recurrent hyperthermia, infection, and hypoglycemia, as well as dysfunctions of the heart, gastrointestinal tract, lungs, kidneys, ocular fundus, and the coagulation system. Despite given ventilator-assisted ventilation, anti-infection therapy, abdominal puncture and drainage, and blood transfusion, the patient still had an aggravated condition and eventually died of multiple organ failures 192 d after birth. Genetic analysis showed that the nuclear family carried compound heterozygous mutations in the COG6 gene (NM_020751.2), including missense mutations of c.662C>T(p.T221M) in exon 7 and c.443T>C(p.I148T) in exon 5, which were both novel mutations and originated from the mother and father, respectively. (2) Literature review: Eight related papers were retrieved, including 20 cases. The main manifestations were various degrees of nervous system abnormalities and growth retardation, complicated by abnormalities of the liver, heart, gastrointestinal tract, blood, immunity, teeth, and bones. All the reported cases suffered from mental and growth retardation, and nine deaths were reported. A total of 11 COG6 gene mutations were identified, and most of them were c.1167-24A>G splicing mutations in a deep intron (seven cases), followed by c.1646G>T (four cases) and c.511C>T (three cases). Conclusions:COG6-CDG commonly manifests as multi-system and multi-organ dysfunctions with poor prognosis. Gene detection is conducive to the accurate diagnosis of COG6-CDG. Our case carries compound heterozygous mutations of c.662C>T(p.T221M) and c.443T>C(p.I148T), which are unreported novel mutations.

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 compare classical and molecular typing methods for Streptococcus pneumoniae,in order to provide fasty and accurate method for classification.Methods Selected the 150 strains of Streptococcus pneumoniae which isolated from the First People’s Hospital of Zhaoqing from October 2013 to February 2015 for the research objects,using the classi-cal quellung classification method and molecular typing of multiple PCR method for classification.Compared the result and the rate of the classification,used the non parametric test of two independent samples for statistical analysis several higer rate of serotypes,and calculated the total coincidence rate,sensitivity and specificity.Results The typing rate of Streptococ-cus pneumonia capsular was 53.3%,and the main serum group were 19(32.7%),6(7.3%)and 23(4%)respectively.The serotype of 19F(23.3%),19A(2.7%),6C(2%)and 23F(2.7%)respectively by single factor serum.The typing rate of the multiple PCR method was 76.0%,and the main serum group were 19(41.3%),6(10.7%)and 23(10.7%).The serotype were mainly 19F(32.7%),19A(8.0%),6(10.7%)and 23F(9.3%).Used the 2 Independent Sample Test for statistical a-nalysis,compared with the serotypes 19F,19A and 23F(P>0.05).There was no statistically significant difference in the distribution of the types of the two methods.The total coincidence rate was 74.6%.To verify the classic quellung method by multiple PCR method,sensitivity was 70.2%,and specificity was 100%.That showed the capsular swelling method was less sensitivity than the molecular typing method.Conclusion Compared with the traditional serological typing techniques,mo-lecular typing technique have the advantages of fast,accurate,sensitive and high resolution,and become the classification method which can be rely on.