Innovative Teaching of "Doctor-patient" Friendly Model for Diabetes from the Perspective of Deep Blended Learning / 中国医学伦理学

The teaching of diabetes has always been the focus and difficulty of clinical internal science teaching. How to skillfully integrate theoretical knowledge into clinical practice and help students deeply master medical knowledge and clinical skills is a topic that clinical teaching teachers are considering. Under the background of analyzing the concept and connotation of deep blended learning, this paper constructed the diabetes "doctor-patient" friendly model teaching process in the perspective of deep blended learning from the three stages of "learning situation analysis and independent learning" in the early stage, "ability improvement and in-depth research" in the middle stage, and "reflection and ability expansion" in the later stage. This paper expounded the positive role of the "doctor-patient" friendly model of diabetes in clinical teaching from three main aspects: improving the integration effect of theory and practice, fully meeting students’ learning needs, and increasing the connection between doctors, patients, and students.

Survey on post competency of general practitioners with completion of residency training / 中华全科医师杂志

Objective:To survey the post competency of general practitioners who completed residency training in Suzhou Municipal Hospital.Methods:A questionnaire survey on post competency of general practitioners was conducted from January to March 2022. General practitioners who completed standardized residential training in Suzhou Municipal Hospital from 2009 to 2021 were randomly selected for the survey. The self-designed questionnaire included the basic information and post competency in terms of clinical, public health, research and teaching abilities as well as medical ethics and humanism. A self-evaluation was also performed and the abilities were graded (A 86-100, B 70-85, C 55-69, D 0-54), and grade A was classified as excellent.Results:A total of 163 questionnaires were distributed and 157 valid ones were collected with a recovery rate of 96.3%. Among 157 respondents 62 (39.5%) were males. The participants mainly worked in urban community health service institutions (78 (49.7%)), and most of them worked as general practice (119 (75.8%)). For the self-assessment of clinical skills, the proportion of respondents with excellent abilities in history taking, basic drug use, diagnosis and treatment of common diseases, and chronic disease management was 58.8% (70/119), 57.1% (68/119), 54.6% (65/119) and 54.6% (65/119), respectively. The proportion of respondents with excellent abilities in evidence-based clinical decision making, physical examination, tests interpretation, referral services, family medical services, rehabilitation services, first aid, and psychological counseling and treatment was 43.7% (70/119), 42.9% (52/119), 38.7% (46/119), 37.8% (45/119), 33.6% (40/119), 22.7% (27/119), 21.0% (25/119), and 16.8% (20/119), respectively. For the self-assessment of basic public health service ability, the proportion of respondents with excellent abilities in health education, disease prevention and control, health management, health care for key and special groups, handling public health emergencies, management of infectious diseases, epidemiology-based community diagnosis and community health leadership was 38.7% (46/119), 33.6% (40/119), 33.6% (40/119), 26.1% (31/119), 25.2% (30/119), 2.7% (27/119), 22.7% (27/119), and 21.0% (25/119), respectively. For humanistic literacy, the proportion of respondents with excellent abilities in privacy protection, sense of responsibility for patients, understanding patients′ needs, effective communication and cooperation with patients was 82.4% (98/119), 73.9% (88/119), 61.3% (73/119), 55.5% (66/119) and 2.1% (62/119), respectively. For research and teaching, the proportion of respondents with excellence abilities in continuous learning and innovation, training and teaching and literature retrieval was 47.9% (57/119), 10.9% (13/119), 10.1% (12/119), respectively. In addition 56.3% (67/119) of respondents were interested in scientific research, 23.5% (28/119) had published articles as the first author or correspondence author, and only 6.7% (8/119) had scientific research projects in the last 5 years.Conclusion:The post competency of general practitioners who received standardized residency training in our hospital varies in different aspects, their abilities in basic public health service, scientific research and teaching are relatively low, which need to be strengthened.

A high-efficiency and versatile CRISPR/Cas9-mediated HDR-based biallelic editing system / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9), the third-generation genome editing tool, has been favored because of its high efficiency and clear system composition. In this technology, the introduced double-strand breaks (DSBs) are mainly repaired by non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways. The high-fidelity HDR pathway is used for genome modification, which can introduce artificially controllable insertions, deletions, or substitutions carried by the donor templates. Although high-level knock-out can be easily achieved by NHEJ, accurate HDR-mediated knock-in remains a technical challenge. In most circumstances, although both alleles are broken by endonucleases, only one can be repaired by HDR, and the other one is usually recombined by NHEJ. For gene function studies or disease model establishment, biallelic editing to generate homozygous cell lines and homozygotes is needed to ensure consistent phenotypes. Thus, there is an urgent need for an efficient biallelic editing system. Here, we developed three pairs of integrated selection systems, where each of the two selection cassettes contained one drug-screening gene and one fluorescent marker. Flanked by homologous arms containing the mutated sequences, the selection cassettes were integrated into the target site, mediated by CRISPR/Cas9-induced HDR. Positively targeted cell clones were massively enriched by fluorescent microscopy after screening for drug resistance. We tested this novel method on the amyloid precursor protein (APP) and presenilin 1 (PSEN1) loci and demonstrated up to 82.0% biallelic editing efficiency after optimization. Our results indicate that this strategy can provide a new efficient approach for biallelic editing and lay a foundation for establishment of an easier and more efficient disease model.

Effects of ruminative thinking on 1-year outcomes in patients with first-episode depression: the mediating role of family functioning / 中华行为医学与脑科学杂志

Objective:To investigate whether rumination and family functioning can predict the level of depression after 1 year of follow-up in patients with first-episode depression, and whether family functioning plays a mediating role between rumination and depression level.Methods:Sixty-five patients with first-episode depression who met the enrollment requirements were included, and all subjects were assessed the 17-item Hamilton depression scale(HAMD-17), rumination response scale(RRS) and family assessment device(FAD). All subjects were followed up for 1 year, and the predictive effects of rumination and family functioning at baseline on the level of depression after 1 year of follow-up were investigated by hierarchical linear regression analysis and mediation analysis.Results:At the baseline stage, rumination, role, affective involvement (AI) and general functioning (GF) were significantly positively associated with depression level after 1 year of follow-up in patients with first-episode depression ( r=0.49, P<0.01; r=0.30, P=0.02; r=0.43, P<0.01; r=0.50, P<0.01; respectively). Rumination, AI and GF at the baseline stage predicted depression level after 1 year of follow-up ( β=0.315, t=2.954, P=0.005; β=0.261, t=2.550, P=0.013; β=0.323, t=2.952, P=0.005). Mediation analysis showed that AI and GF partially mediated the relationship between rumination at baseline and depression level at 1 year follow-up (point estimate value for AI=0.040, 95% CI=0.012-0.090); point estimate value for GF=0.066, 95% CI=0.017-0.143). Conclusions:Rumination and family functioning at baseline in first-episode depressed patients can predict the depression level at 1 year follow-up.Family functioning partly mediates the relationship between the baseline rumination and the depression level at 1 year follow-up.

Relationship between classroom lighting and poor vision of students in primary and secondary schools in Tianjin / 中国学校卫生

Objective@#To investigate the relationship between classroom lighting and poor vision of students in primary and secondary schools in Tianjin, and to provide reference for targeted prevention measures for comprehensive school eye health programmes.@*Methods@#A total of 623 classrooms and 24 713 students in 89 primary and secondary schools in Tianjin were selected using stratified cluster sampling method in Sep.to Oct. of 2020. The illuminometer was used to monitor the lighting environment of the classroom, and the standard logarithmic vision light box was used to detect the naked eye vision of students. Chisquare test, variance analysis and binary Logistic regression were used in SPSS software.@*Results@#Rate of low vision among primary and secondary school students in Tianjin was 67.86%. The qualification rate of blackboard reflectance, blackboard average illumination, blackboard illumination uniformity, desk average illumination and desk illumination uniformity were 58.11%, 53.13%, 73.19%, 66.61% and 75.12%, respectively. Univariate analyses showed that the blackboard reflectance, blackboard average illumination, desk average illumination, and desk illumination uniformity were associated with low vision among students(χ 2=311.29, 62.54, 61.71, 6.59, P<0.05). Multivariable Logistic regression analysis showed that blackboard reflectance, average illumination of blackboard and desk were associated with higher risk of poor vision[OR(95%CI)=1.19(1.11-1.27),1.27(1.17-1.37),1.11(1.02-1.20), P<0.05].@*Conclusion@#Prevalence of low vision among primary and secondary school students in Tianjin is relatively high. Blackboard reflectance, average illumination of blackboard and desk are the important determinants for poor vision of primary and secondary school students. Creating effective classroom lighting scheme is crucial for vision health among students.

A case of maternal 45, X/46, XX mosaicism detected by non-invasive prenatal testing / 中华医学遗传学杂志

Objective@#To report on a case of maternally derived 45, X mosaicism detected by non-invasive prenatal testing (NIPT).@*Methods@#Fetal sex chromosomal abnormality was detected by NIPT. Maternally derived 45, X mosaicism was confirmed by chromosome karyotype analysis. Fetal sex chromosome aneuploidy was detected by amniotic fluid chromosome microarray analysis.@*Results@#A maternal 45, X mosaicism was diagnosed. The fetus was confirmed to be normal.@*Conclusion@#Maternal 45, X masaicism can be diagnosed by NIPT.

A case of maternal 45,X/46,XX mosaicism detected by non-invasive prenatal testing / 中华医学遗传学杂志

OBJECTIVE@#To report on a case of maternally derived 45,X mosaicism detected by non-invasive prenatal testing (NIPT).@*METHODS@#Fetal sex chromosomal abnormality was detected by NIPT. Maternally derived 45,X mosaicism was confirmed by chromosome karyotype analysis. Fetal sex chromosome aneuploidy was detected by amniotic fluid chromosome microarray analysis.@*RESULTS@#A maternal 45,X mosaicism was diagnosed. The fetus was confirmed to be normal.@*CONCLUSION@#Maternal 45,X masaicism can be diagnosed by NIPT.

Prognosis of patients with coronary heart disease and normal myocardial perfusion imaging / 心血管康复医学杂志

Objective: To observe influence of different risk factors on prognosis of patients with coronary heart disease (CHD) and normal myocardial perfusion imaging outcome.Methods: A total of 99 CHD patients with normal myocardial perfusion imaging outcome were selected.Left ventricular function indexes were measured by gated resting myocardial imaging in resting and stress state.All patients received telephone follow-up until natural death (died of other causes) or fatal or non-fatal heart attacks, or the termination of the experiment after 45 months.Cox proportion risk regression model was used to analyze risk factors of fatal and non-fatal heart attacks.Results: A total of 15 cases died during the 45-month follow-up.Mean all-cause mortality per year was 5.05%.Fatal heart attacks occurred in nine cases (9.09%), and non-fatal heart attacks occurred in 21 cases (21.21%).Cox proportion risk regression analysis indicated that smoking and left ventricular ejection fraction (LVEF) <50% were risk factors for fatal heart attacks (HR=4.887, 3.365, P=0.043, 0.002), while diabetes mellitus, dyslipidemia, smoking and LVEF<50% were risk factors for non-fatal heart attacks (HR=2.215~4.544, P<0.05 all).Conclusion: Incidence rate of cardiovascular events is higher in CHD patients with normal myocardial perfusion imaging.Smoking and impaired heart function suggest poor prognosis in these patients.

Relationship between blood pressure variability and left ventricular diastolic function in patients with ;essential hypertension / 心血管康复医学杂志

Objective:To observe the relationship between dynamic blood pressure variability (BPV) and left ventric‐ular diastolic function in patients with essential hypertension (EH) .Methods :A total of 140 newly diagnosed EH pa‐tients were selected .According to total coefficient of variation (CV) of 24h systolic blood pressure BPV ,they were divided into high CV group (n=70 ,CV>12.16% ) and low CV group (n=70 ,CV≤12.16% ) .Another 70 healthy subjects with corresponding gender and age were enrolled as healthy control group simultaneously .Echocardiography was used to measure mitral early diastolic peak flow velocity (E) ,late diastolic peak flow velocity (A) and decelera‐tion time of E peak (DT);meanwhile ,tissue Doppler mode was used to record early diastolic peak velocity (Em) and late diastolic peak velocity (Am) .All above indexes were compared among all groups .Results:Compared with healthy control group ,BPV significantly rose in EH patients (P<0.05 or < 0.01) .All BPV indexes in high CV group were significantly higher than those of low CV group except daytime mean diastolic blood pressure variability (dDBP‐BPV) and nighttime mean diastolic blood pressure variability (nDBP‐BPV) , P<0.05 all .Compared with healthy control group ,there were significant reductions in E and Em/Am ,and significant rise in DT and E/Em in EH patients ,P<0.05 or <0.01 ;compared with low CV group ,there was significant rise in E/Em [(10.32 ± 3.20) vs .(14.22 ± 2.20) ,P=0.033] in high CV group .Conclusion:Left ventricular diastolic dysfunction is more severe in hypertensive patients with higher dynamic blood pressure variability .

Formulation and application of drainage tube nursing program in step-up mini-invasive surgery for infected pancreatic necrosis / 中华护理杂志

This paper summarized drainage tube nursing experience instep-up mini-invasive surgery for infected pancreatic necrosis,special management group for drainage nursing was established,and drainage tube management in stages was implemented. Nursing key pointsin the first stage was: catheter puncture assistance,single-site and single-tube fixed drainage,anddynamicdrainage effect observation. Nursing key points in the second stage was:single-site and multi-tube fixed drainage,close surveillance for negative pressure drainage effect,and strengthened protection of drainage tubeduring patient activities. Nursingkey points in the third stage was: multi-site and multi-tube fixed drainage,prevention of hospital-acquired infection,and discharge guidance for patientsin drainage tube care. The cure rate of this group was 91.93% with 62 patients,and there was no occurrence of unplanned extubationduring hospitalization.

Evaluation of the Xpert MTB/RIF assay for diagnosis of tuberculosis and rifampin resistance in county-level laboratories in Hunan province, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The Xpert MTB/RIF showed high sensitivity and specificity in previous studies carried out in different epidemiological and geographical settings and patient populations in high-burden tuberculosis (TB) countries. However, there were little data obtained by validation or demonstration study of the assay in China. In this study, the performance of Xpert MTB/RIF was investigated in two county-level laboratories in Hunan Province, China.</p><p><b>METHODS</b>Consecutive patients with suspected pulmonary tuberculosis (PTB) and suspicion for multidrug-resistant tuberculosis (MDR-TB) were enrolled. For each patient suspected to have PTB, three sputum specimens (one spot sputum, one night sputum, and one morning sputum) were collected and each sputum was tested with smear microscopy, Löwenstein-Jensen (LJ) culture, and Xpert MTB/RIF test. For comparison across subgroups and testing methods, 95% confidence intervals were calculated. All analyses were done with SPSS 16.0, and P < 0.05 was regarded as significant.</p><p><b>RESULTS</b>For case detection, the sensitivity of Xpert MTB/RIF was 100% for smear- and culture-positive TB and 88.6% for smear-negative and culture-positive TB; the overall sensitivity was 94.5% for all culture-positive patients. The specificity was 99.8%. The sensitivity of Xpert MTB/RIF assay was 22.0% in clinical TB patients and the specificity reached 100.0% in the group of patients who are infected with nontuberculous mycobacteria. For the detection of rifampin resistance, the sensitivity of MTB/RIF RIF-resistance detection was 92.9%, and the specificity was 98.7%. Of the 26 Xpert MTB/RIF-positive and RIF-resistant patients confirmed by LJ proportion tests, 20 (76.9%) patients were infected by MDR-TB.</p><p><b>CONCLUSIONS</b>The Xpert MTB/RIF assay is a highly sensitive and specific method for diagnosis of TB and RIF resistance, which will enable it to have the potential to be used in county-level laboratories and lead to the reduction of the infectious pool and improvements in TB control in China. Further evaluations in county-level laboratories for implementing the assay are still required.</p>

The feasibility study of Ion Proton semiconductor sequencing platform in the non-invasive prenatal genetic diagnosis / 中华检验医学杂志

Objective To evaluate the feasibility of apply Ion Proton semiconductor sequencing platform in non-invasive prenatal genetic diagnosis .Methods Totally 1 000 pregnant women with a singleton pregnancy of 12-32 weeks gestation were selected from the Third affiliated Hospital of Zhengzhou University from Jan to Dec 2013.Using noninvasive prenatal genetic diagnosis based on Ion Proton semiconductor sequencing platform to study their cffDNA .In parallel, 72 pregnant women received invasive prenatal diagnosis by traditional chromosomal analysis with amniocentesis chorionic villus sampling .Results It′s shown that 18 out of 1 000 (1.8%) pregnant women underwent the noninvasive prenatal genetic testing had a high risk for aneuploid chromosomes , including 7 cases of 21-trisomy, 4 cases of 18-trisomy, 2 cases of 13-trisomy, 4 cases of sex chromosomal abnormality , and 1 case of 15-trisomy.It demonstrated that the rate and accuracy of fetal 21-trisomy, 13-trisomy and 18-trisomy by non-invasive prenatal genetic testing were both 100%without misdiagnosis , the rate of detection for sex chromosomal abnormality was 2/2 with a false positive rate of 1/3.However, the 15-trisomy predicted by the non-invasive prenatal diagnosis in a woman was finally proved to be a false positive .Based on the results by karyotyping (55/55) as well as follow-ups (493/493), the specificity of the non-invasive prenatal diagnosis for detection of 21-trisomy, 18-trisomy and 13-trisomy was 100%.One Ion PITM chip could detect 12 to 15 samples in 1.5 h and the whole process of noninvasive detection could be completed in 1 to 1.5 days.Conclusions The non-invasive prenatal diagnosis by Ion Proton semiconductor sequencing platform could provide fast and accurate detection of fetal aneuploidy .The benchtop high-throughput sequencing platform has laid the foundation for the independent application in clinical settings for fetal aneuploidy detection .

Role and mechanism of retinoic acid in axonal regeneration / 中国组织工程研究

BACKGROUND:Retinoic acid signaling pathways is very important in the formation pf nervous system, specialization of neurons and outgrowth of axons. The recent studies show that, retinoic acid plays an important role in the process of axonal regeneration, but few research reports its exact molecular mechanism. OBJECTIVE:To analyze and summarize the mechanism underlying retinoic acid signaling pathways in the process of axonal regeneration. METHODS:A computer-based online research was conducted among the VIP, CNKI, PubMed, BioMed Centeral, Springer, The Free Medical Journals, EBSCO and Foreign Journals Integration System between January 2000 and December 2013, with the key words of“retinoic acid, the central nervous system, nerve damage, axon regeneration, and mechanism”in Chinese and English. A total of 43 studies addressing the molecular mechanism of retinoic acid in axonal regeneration were screened. According to the supplementary documents, another five references were added. Repetitive research and atypical reports were excluded. RESULTS AND CONCLUSION:Fol owing acute central nervous system injury, axonal regeneration and functional recovery are extremely limited. For proper functionality fol owing injury, axons must regrow, reinnervate their targets, and remyelinate their axons. When the central nervous system injuries occur, retinoic acid signaling pathways express transcription factor retinoic acid receptorβ2 to induce axonal regeneration fol owing injury;in dorsal root ganglion neurons, cAMP levels are greatly increased by lentiviral retinoic acid receptorβ2 expression and contribute to neurite outgrowth. More recently, retinoic acid-retinoic acid receptorβ2 pathways directly transcriptional y repress a member of the inhibitory Nogo receptor complex, Lingo-1, under an axonal growth inhibitory environment in vitro as wel as fol owing spinal cord injury in vivo. Through these molecular mechanisms, retinoic acid signaling pathways play its important role in the process of axonal regeneration.

The treatment of Candesartan joint CSDP on type 2 diabetic nephropathy with stage Ⅲ / 中国综合临床

Objective To explore the clinical effect of CSDP combined with losartan treatment on type 2 diabetic nephropathy with stage Ⅲ.Methods Sixty patients suffered from type 2 diabetic nephropathy with stage Ⅲ were recruited as our subjects.They were randomly divided into Candesartan group and Candesartan + CSDP group,each for 30 patients.Patient in Candesartan + CSDP group were administrated with candesartan 4 mg,once per day,the CSDP 15 capsules and three times / day for 3 months.Patients in Candesartan group were given:candesartan 4 mg once/day for 3 months.The level of urinary albumin,serum urea nitrogen (BUN) and creatinine (Cr) were measured of subjects before and after treatment.Results The levels of BUN,Cr were lower after any treatment than before treatment in 2 groups.The difference of BUN,Cr in Candesartan + CSDP group were(-14.6 ± 8) mg/L,(-11.9 ± 3.3) μmmol/L,(-0.7 ± 0.2) mmol/L,higher than that of Candesartan group (-7.1 ± 7.8) mg/L,(-8.5 ± 0.8) μmol/L,(-0.4 ± 0.1) mmol/L] (P ＜ 0.01).Conclusion The treatment strategy of Candesartan joint CSDP can be method regarding of reducing urinary albumin and BUN,Cr in patients of type 2 diabetic nephropathy with stage Ⅲ.

Effect of noninvasive positive pressure ventilation on the severe cases of hand foot and mouth disease / 中国小儿急救医学

Objective To evaluate the effects of noninvasive positive pressure ventilation on the severe cases of hand foot and mouth disease (HFMD).Methods Eighty-six cases of severe HFMD admitted to our hospital between Apr 2009 and Aug 2010 were divided into two groups based on the time when admitted to our hospital.Thirty-eight cases (group A,Apr to Jul,2009) received routine treatment and 48 cases (group B,Apr to Aug,2010) were ventilated with nasal continuous positive airway pressure based on routine treatment.The difference of the total efficiency was compared between two groups.Results Twenty-five out of 38 patients were cured,six showed improved signs,and seven showed no improvement in group A,of which the total efficiency was 81.5％.Forty out of 48 patients were cured,seven showed improved signs,and one showed no improvement in group B,of which the total efficiency was 97.9％.The clinical effect of group B was much better than group A and the difference was statistically significant ( x2 =6.71,P ＜0.05).Conclusion Early nasal continuous positive airway pressure has most valuable effect on HFMD and is helpful in preventing -HFMD from getting worse.

Screening of infection of malaria parasite with, atypical eosinophil distributions in the WBC scattergram and alarm in Sysmex XE-2100 hematology analyzer / 中华检验医学杂志

Objective To explore a quick and feasible method of screening malaria parasite by using a Sysmex XE-2100 hematology analyzer though alarm information on high eosinophil count and atypical eosinophil distributions in the WBC scattergram. Methods Sysmex XE-2100 hematology analyzer was used for complete blood cell analysis. Microscopic review was used when high eosinophil count and atypicaleosinophil distributions in the WBC scattergram were found. If the review showed normal eosinophil cells, wecontinued to focus on red cell for searching malaria parasite. Results Among 1 501 specimens showing higheosinophil counts and atypical eosinophil distributions, nine cases with normal eosinophil cells were indisaccordance with the hematology analyzer, six of them showed high eosinophil count in the Sysmex XE-2100 hematology analyzer, whose distribution was located close to neutrophil clusters in scattergram. The otherthree had an abnormal WBC scattergram. There was no gap between eosinophil clusters and neutrophilclusters, which brought no classified results. But all the nine specimens had been found the trophozoite,schizont and merozoite in blood smears. Conclusions There were great possibility of the existence of themalaria parasite in specimens when hematology analysis showed high eosinophil count and atypical eosinophildistributions in the WBC scattergram in a Sysmex XE-2100 hematology analyzer, although these alarm wasnot comfirmed by microscopic review. This method provides not only a simple and reliable way for quickscreening malaria parasite but also has a great value in preventing the undetected ratio on malaria parasite.

Identification of neural tissue-committed stem cells, a new cellular group, in bone marrow / 中国组织工程研究

BACKGROUND: It has been widely accepted that both bone marrow-derived mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) have the capacity to differentiate into neural lineages. Some scholars believe that in addition to HSCs and MSCs, bone marrow (BM) also harbors a highly mobile population of CXCR4+ tissue committed stem cells (TCSCs), including skeletal muscles, heart, liver, and neural tissue. OBJECTIVE: To make sure that neural tissue-committed stem cells (NTCSCs) reside in the bone marrow, and to establish a purification and culture method for bone marrow-derived NTCSCs.DESIGN: Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA. MATERIALS: Adult Sprague-Dawley (SD) rats (pathogen-free) were provided by the Animal Center of the Second Military Medical University of Chinese PLA. Dulbecco's modified eagle's medium (DMEM)/F12, B27, N2 and epidermal growth factor (EGF, Invitrogen Company), basic fibroblast growth factor (bFGF, CytoLab Ltd), rabbit anti-rat Nestin,CXCR4, β-Tublin Ⅲ, glial fibrillary acidic protein (GFAP, Santa Cruz Company), mouse anti-rat microtubule associated protein 2ab (MAP2ab) (Clone11-5B), cyclic nucleotide 3'phosphohydrolase (CNPase, Clone AP20, NeoMarkers Company), fluorescent(fluorescein isothiocyanate, Cy3) marker reagents (Wuhan Boster Bioengineering Co., Ltd), nuclear fluorescent dyes 4,6-diamidino-2-phenylindole(DAPI)(Sigma), immunohistochemistry reagents (Vector Laboratories Company) , and NycoPrepTM separation liquid (1.077A, Axis-Shield Company) were used in this study.METHODS: This study was performed in the Department of Anatomy, the Second Military Medical University of Chinese PLA from January 2004 to December 2006. Bone marrow was harvested from bilateral femurs and tibias of 2-3 weeks SD rats. Mononuclear cell layer was isolated by NycoPrepTM separation liquid and suspended in DMEM/F12(1:1)serum-free medium supplemented with 2% B27,1% N2, 20 μg/L bFGF, 20μg/L EGF, 1×105 U/L penicillin and 100 mg/L streptomycin. NTCSCs were isolated and propagated by suspensive growing from adherent cells in bone marrow in DMEM/F12 free-serum medium. MAIN OUTCOME MEASURES: NTCSCs were identified by immunocytochemistry for CXCR4, a marker of TCSCs and nestin, a marker of neural stem cells, and neural lineages marker protein after differentiation of cellular spheres. RESULTS: The NTCSCs spheres expressed nestin, a neural stem cell marker as well as CXCR4, a marker of TCSCs. The NTCSCs' spheres were naturally differentiated in DMEM medium with 15% fetal bovine serum. The differentiated cells expressed β-Tublin Ⅲ, MAP2ab, CNPase and GFAP, markers of neural lineages. CONCLUSION: NTCSCs reside in bone marrow and naturally differentiate into neural lineages in vitro.

Clinical evaluation of interventional treatment of severe complicated stenosis of vertebrobasilar system / 介入放射学杂志

Objective To evaluate the feasibility,safety and efficacy of percutaneous transluminal stenting angioplasty for severe complicated stenosis of vertebrobasilar system. Methods From November 2003 to February 2006,5 candidates underwent percutaneous transluminal stenting for severe complicated stenosis of vertebrobasilar system. Results Four out of 5 candidates,had occlusion of unilateral vertebral artery (VA),1 had severe bilateral proximal segmental stenosis of VA. There were 4 with severe proximal segmental stenosis of the VA associated with multiple segmental stenosis of intracranial VA and basilar artery (BA),and 1 with multiple segmental severe stenosis of intracranial VA and BA. Stenosis rate ranges from 80% ～95% with involved length from 10-20 mm. Technical success was achieved in all of the patient (100%),and residual stenosis rate was less than 20%. All the symptoms due to vertebrobasilar blood supply insufficiency disappeared. Follow-up with DSA 6-12 months later demonstrated no restenosis; showing satisfactory short term efficacy. Conclusions Percutaneous transluminal stenting for vertebrobasilar blood supply insufficiency is a safe and efficacious option with favorable short term outcome,especially with furthermore prevention of stroke.

Electro-acupuncture on Du meridian up-regulates the expression of Bcl-2 mRNA and its protein in early spinal cord injury of rats / 中国组织工程研究

BACKGROUND: It is proved that acupuncture can remarkably promote recovery of nervous function after spinal cord injury (SCI). Previous studies in our groups have been proved that electro-acupuncture can inhibit apoptosis in early period of SCI, but the mechanism is unclear yet.OBJECTIVE: To study the effect of electro-acupuncture on expressions of apoptosis inhibitory gene Bcl-2 mRNA and protein with hybridization in situ and immunohistochemistry and discuss the possible mechanism of apoptosis inhibited by electro-acupuncture in early SCI.DESIGN:Opening animal study.SETTING: Department of Anatomy, the Second Military Medical University of Chinese PLA; Shanghai Acupuncture & Moxibustion and Meridian Research Center.MATERIALS:Adult male SD rats of pathogen-free aged 2-3 months were selected in this study. Bcl-2 hybridization in situ kit was provided by Wuhan Boshide Biotechnology Company Limited and Bcl-2 antibody (1:200) was provided by Santa Cruz Biotechnology Company.METHODS: The experiment was completed at the Laboratory of Anatomy of the Second Military Medical University of Chinese PLA from July 2004 to December 2005. All experimental rats were randomly divided into model group, electro-acupuncture group, methylprednisolone group and sham operation group. T10 spinal cord was injuried by the modified Allen's method and treated with electro-acupuncture immediately, and then the expressions of Bcl-2 mRNA and protein were evaluated with hybridization in situ and immunohistochemistry combined with image quantitative analysis.MAIN OUTCOME MEASURES: ① Expressions of Bcl-2 mRNA and protein in early SCI; ② effect of electro-acupuncture on expressions of Bcl-2 mRNA and protein.RESULTS: The rats were supplied when they died during the experiment,and all 42 rats were involved in the final analysis. ① Moderate expression of Bcl-2 mRNA and protein was observed in the sham operation group.Expression of Bcl-2 mRNA was increased in model group at 6 hours after SCI, but expression ofBcl-2 protein was not changed. At 24 hours after SCI, both expressions of Bcl-2 mRNA and Bcl-2 protein were increased.Expression of Bcl-2 mRNA and protein was higher in electro-acupuncture group than that in mo del group (P ＜ 0.05), and there was no significant difference from that in methylprednisolone group. ② Amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and methylprednisolone group at 6 hours after treatment, and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01). After 24-hour treatment, amount of positive Bcl-2 mRNA cells was increased in electro-acupuncture group and gray value was decreased.There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01). ③ After 24-hour treatment, amount of immunohistochenistry positive Bcl-2 cells was increased in electro-acupuncture group and gray value was decreased. There was significant and remarkably significant difference from those in sham operation group and model group, respectively (P ＜ 0.05-0.01).CONCLUSION: Electro-acupuncture can up-regulate the expressions of Bcl-2 mRNA and protein so as to inhibit apoptosis in early SCI.

Study on mRNA expression of P2y receptors of sciatic nerve in rate / 中华显微外科杂志

Objective To demonstrate the presentation of the extracellula ATP receptors in peripheral nerve in rats Methods Reverse transcriptase polymerase chain reaction was performed in sciatic nerve of rat Results There were three P2y receptor mRNA expressions in the in the sciatic nerve of the rat,they were P2y2 and P2y4 as well as P2y6 Conclusions Extracellular ATP,as a neurotransmitter,activates the P2y purinoceptors located on the axon regeneration after peripheral nerve injury and decided the direction of the regeneration axon The P2y receptors are related to the regeneration of the peripheral nerve