Advance in molecular genetic research on primary congenital glaucoma / 中华医学遗传学杂志

Primary congenital glaucoma (PCG) is one of the major diseases causing blindness in children, but its pathogenesis has remained unclear. Genetic factors play an important role in the pathogenesis of PCG. Molecular genetics of candidate genes such as CYP1B1, MYOC, LTBP2 and FOXC1 has so far been explored, but no disease-causing gene has been identified. Molecular genetic research on PCG including candidate gene screening and research strategies are reviewed here.

Manufacture and evaluation of integrated biphasic silk fibroin scaffold made by annulus fibrosus-nucleus pulposus tissue engineering / 天津医药

Objective To assess the prospect of integrated biphasic silk fibroin scaffold made by annulus fibrosus-nu?cleus pulposus tissue engineering in application as integrated intervertebral disc(IVD). Methods An integrated annulus fi brosus-nucleus pulposus(AF-NP)biphasic scaffold was made by silk fi broin using two different uncomplicated methods which were paraffin spheres-leaching method(outer AF phase)and phase separation method(inner NP phase). The scaf?fold was investigated by general observation, stereomicroscope and scanning electron microscopy(SEM). Its pore size, poros?ity, and compressive elastic modulus were determined. AF and NP cells were isolated from rabbit IVD and seeded into the corresponding phase of the scaffold respectively. The cell-scaffold complex was cultured for 48 hours. The biocompatibility of the scaffold was evaluated by SEM, live/dead staining while CCK-8 assay was used to assess cell proliferation. Results Stereomicroscope and SEM showed that AF phase and NP phase integrated perfectly without cross-linking. Both phases pos?sessed highly interconnected porous structure [pore size of AF and NP phase were(220.0±23.1)μm and(90.0±17.8)μm, re?spectively] and highly porosity(AF and NP phase were respectively 91%and 93%). In addition, this silk biphasic scaffold had impressive mechanical properties(150.7 ± 6.8)kPa. SEM revealed that disc cells attached to regions of pore walls, dis?tributed uniformly and secreted extracellular matrix. Live/Dead staining and cell count kit-8(CCK-8)analysis showed that the silk composite scaffold was non-cytotoxic to disc cells. Conclusion This silk biphasic AF-NP scaffold has satisfied pore size, porosity, biomechanical properties and biocompatibility, so it is ideal candidate for IVD tissue engineering.

Association between LOXL1 gene polymorphisms and primary open angle glaucoma in Sichuan population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate association between the lysyl oxidase-like 1 (LOXL1) gene single nucleotide polymorphism (SNP) and primary open-angle glaucoma (POAG) in Sichuan population.</p><p><b>METHODS</b>In this study,416 subjects with primary open-angle glaucoma and 997 normal controls were recruited.Three reported LOXL1 tag SNPs (rs1048661,rs3825942 and rs2165241) were genotyped by SNaPshot method.</p><p><b>RESULTS</b>The study showed that the genotypes of LOXL1 rs1048661,rs3825942 and rs2165241 between POAG and control groups were not statistically significant (OR=1.085, 95%CI 0.92-1.28, P=0.578 for rs1048661; OR=1.059, 95%CI 0.82-1.37, P=0.846 for rs3825942; OR=1.006, 95%CI 0.77-1.32, P=0.966 for rs2165241, respectively). There were no significant difference in allele frequency distribution of LOXL1 rs1048661、rs3825942 and rs2165241 between POAG and normal controls (P=0.322, P=0.660, P=0.965).</p><p><b>CONCLUSION</b>The results from the present study do not indicate the association of LOXL1 SNPs (rs1048661, rs3825942 and rs2165241) with POAG in Sichuan population.</p>

An Experimental Study on Bovine Nucleus Pulposus Cells Labelled with PKH26 in Vitro / 天津医药

Objective To investigate the application of PKH26 fluorescent labeling on nucleus pulposus cells isolat-ed from bovine coccyx disc, and to provide nucleus pulposus tissue engineering with traceable nucleus pulposus cells by PKH26 fluorescence labelling. Methods Nucleus pulposus primary cells were isolated from the nucleus pulposus tissue de-tached from bovine coccyx disc by enzymatic digestion, and observed under the inverted microscope. Safranin O, toluidine blue and type Ⅱ collagen immunocytochemistry methods used to stain for passage one generation cells. Nucleus pulposus cells were labeled with PKH26 fluorescence in accordance with the instructions. The cell activity, fluorescence intensity at d0, d14 and d28 of culture, characteristics of proliferation and the expression of gene in labeled cells were assessed. Re-sults Isolated nucleus pulposus cells amounted to (1.56 ± 0.35) × 106/g. Under the inverted microscope, primary cells ad-hered at the 4 th day of culture, grew in groups, and covered the bottom of culture flask at the 13 th day. Both primary cells and the P1 generation cells were chondrocyte-like morphology. The staining of safranin O, toluidine blue and typeⅡcolla-gen immunocytochemistry for P1 generation of nucleus pulposus cells showed positive results. The cell activity before and af-ter PKH26 labeling showed more than 95%, and the fluorescence intensity at d0, d14 and d28 performed a decreasing trend, but still showed detect strong fluorescence at d28. There were no significant differences in proliferation and the expression of gene (collagen typeⅠandⅡ, aggrecan) before and after cell labeling (P>0.05). Conclusion As the seed cells of tissue en-gineering, nucleus pulposus cells isolated from bovine coccyx can reach a satisfactory number and maintain cartilage-like phenotype, and no changes shown in the biological characteristics after labeling. PKH26 labeled nucleus pulposus cells are suitable for the traceable cells in vivo study.

Preliminary Construction of Tissue Engineering Nucleus Pulposus Combining Silk Fibroin Porous Scaffold with Rabbit Nucleus Pulposus Cells / 天津医药

Objective To investigate the feasibility of construction of tissue engineering nucleus pulposus by com?bining the novel silk fibroin porous scaffold with PKH26 labeled rabbit nucleus pulposus cells. Methods Rabbit nucleus pulposus cells were isolated and cultured, then the passage 1 nucleus pulposus cells were stained with safranin O and typeⅡcollagen immunohistochemical staining. The isolated rabbit nucleus pulposus cells were labeled with PKH26. MTT assay was used for examining the proliferation of the nucleus pulposus cells before and after labeling. Labeled cells were inoculat?ed in the scaffold, cultured for 4 days and then the cell-scaffold complexes were implanted subcutaneously into nude mice. After 12 weeks of in vivo culture, the cell-scaffold complexes were detected by in vivo imaging technology, H&E staining, toluidine blue staining, safranin O staining and collagen typeⅡimmunohistochemical staining. Results Safranin O stain?ing and typeⅡcollagen immunohistochemical staining of the passage 1 nucleus pulposus cells were positive. The fluores?cence intensity of labeled cell was distributed, and the difference of OD value of nucleus pulposus cells was not statistically significant before and after labeling (P>0.05). The in vivo imaging technique showed a strong fluorescencea in porous scaf?fold. H&E staining of cell-scaffold complexes showed that the scaffolds were filled with a large number of nucleus pulposus cells and large amount of extracellular matrix. Toluidine blue staining, safranin O staining and typeⅡcollagen immunohisto?chemical staining were positive, and large amount of extracellular matrix was secreted around the cells. Conclusion The new silk fibroin porous scaffold with rabbit nucleus pulposus cells in vivo culture formed nucleus pulposus like tissue, which can be used for construction of tissue engineering nucleus.

Changes of endogenous growth factors and their significance after sciatic nerve injury in rabbits / 中华创伤杂志

Objective To investigate variations in bFGF,NGF,and brain-derived neurotrophic factor(BDNF) following sciatic nerve injury in rabbits and the optimum time to perform stem cell transplantation.Methods Sixteen New Zealand rabbits were divided into control group and groups at 3,7,and 14 days postinjury according to the random number table,with 4 rabbits per group.Rabbit models of the sciatic nerve injury were induced by forceps.Structural change of the injured nerve tissues were observed with HE staining.Contents of NGF,bFGF,and BDNF in supernatants of homogenated sciatic nerves were detected by ELISA test.Results Level of bFGF increased slowly postinjury,reached the peak at day 7 (P ＜ 0.05),and then restored to the normal level at day 14 (P ＞ 0.05).Level of BDNF raised quickly postinjury,reached the peak at day 7 (P ＜ 0.05),and then restored to the normal level at day 14 (P ＞ 0.05).Level of NGF increased rapidly postinjury,reached the peak at day 3 (P ＜ 0.05),and then restored to the normal level at day 7 (P ＞ 0.05).Conclusions In the early recovery process after peripheral nerve injury,the nerve tissues regulate the secretion of NGF,bFGF,and BDNF immediately and secretions of these growth factors correlate with the time of injury.Early period (3-7 days) after injury is the best time to perform nerve repairing,nerve transplantation,and stem cell transplantation.

Progress of auxiliary examinations in the diagnosis and treatment of hypertrophic cardiomyopathy / 国际儿科学杂志

Hypertrophic cardiomyopathy is regarded as an inherited cardiac disorder caused by autosomal dominant mutations.It has the remarkable heterogeneity of hereditary capacity,clinical phenotype,clinical course and prognosis,thus the diagnosis and treatment of this disease is challenging.Assistant examinations,such as Doppler tissue imaging,quantitative tissue velocity imaging,tissue strain imaging,cardiac magnetic resonance and late gadolinium enhancement,are important to early diagnose,guide management and judge prognosis for hypertrophic cardiomyopathy.This paper reviews the progresses of assistant examinations in hypertrophic cardiomyopathy.

Effects of Cyclic Tensile Strain on Proliferation of Human Osteoarthritis Chondrocytes / 天津医药

Objective To explore the effects of cyclic tensile strain on the proliferation of cultured human osteoar-thritis chondrocytes. Methods The degenerated human cartilage came from a 66-year-old patient with osteoarthritis. The degree of degeneration was assessed by pathological diagnosis. Under the sterile condition, degenerated chondrocytes were cultured by enzymatic digestion. The toluidine blue staining, safranin O staining, and immunofluorescence staining of colla-genⅡwere used to identify the cultured cells. The third generation cells were seeded on the silicone rubber membrane carri-er, using the EF3200 mechanical tester equipped with BioDynamic bioreactor system, imposed a frequency of 0.25 Hz, stain 0, 5%, 10%and 15%load for 3 h. The proliferation activity was detected by flow cytometry. Results The proliferative in-dex (PI) increased with the magnitude value of cyclic tensile strain. The most significant increase of proliferation index was found in 10%group. Conclusion The proliferation of human osteoarthritis chondrocytes increases under some stress.

Measures of the hospital against public health emergencies / 中华医院管理杂志

Infectious disease hospitals are obliged to cope with public health emergencies such as outbreak of infectious diseases.Such hospitals are required to make early detection,early containment,proper management,reasonable preplans,timely training,targeted drills,reasonable deployment of hospital resources,appropriate protection for hospital staff.General hospitals should also cope with prevention and control of infectious diseases to some extent,and work with infectious disease hospitals hand in hand to better cope with such outbreaks.

Progress in research of preparation methods of super paramagnetic iron oxide and the application in cell marking / 国际生物医学工程杂志

Cell transplantation has great potentials in repairing damaged tissue.If we want to use the cell transplants to treat diseases that respond poorly to the conventional treatment,or provide better treatments,in vivo dynamic tracking is particularly important.The application of superparamagnetic iron oxide (SPIO) improves a common inconvenience of the current invasive tests including immunohistochemical study or transmission electron microscopy (TEM) study and so on.Researchers has established a variety of preparation methods of the particles,groped the optimal condition of cell marking.SPIO were proved to be feasible and superior in cell tracking in vivo through animal experiments.The results provide the reliable means of using SPIO to track cell in clinical treatments.This review gives a summary of the related study.

Safety and clinical application of induced pluripotent stem cells / 中国组织工程研究

BACKGROUND: Induced pluripotent stem cells are obtained from somatic cells by reprogramming method. The safety of induced pluripotent stem cells has attracted much attention because of their huge and potential value in clinical application. OBJECTIVE: To review the current studies addressing the safety and clinical application of induced pluripotent stem cells. METHODS: The PubMed database between 2006 and 2012 was retrieved by the first author to search the correlative documents concerning the safety and clinical application of induced pluripotent stem cells. Total y 203 papers were primarily gotten. Final y, 47 papers were included. RESULTS AND CONCLUSION: At present, the main methods to enhance the safety of induced pluripotent stem cells include avoiding usage of c-Myc gene, another mediate way replacing the retrovirus, direct leading of reprogramming factor protein, safer donor cells, micromolecule compound and other in-transgenosis ways. Induced pluripotent stem cells have extensive clinic treatment prospects, and can be used for the build of disease-specific induced pluripotent stem cells line.

Research progress of adipose-derived stem cells in osteochondral tissue engineering / 国际生物医学工程杂志

With the rapid development of medical sciences in recent years,tissue engineering technology has made great achievements,while the choosing of seed cells has been a major focus of the field.Adipose-derived cells have the advantages of rapid expansion,good stability,no immune rejection and potential of multiplex differentiation.They can differentiate into cells originated from different germ layers such as:adipocytes,osteoblasts,chondrocytes,endothelial cells,myocytes,neuronal cells and so on.Besides,adipose tissue can be harvested easily and substantially with small trauma to human body.So ADSCs are expected to be an ideal choice of seed cells in tissue engineering.This article introduces the research progress of ADSCs in osteochondral tissue engineering.

Relationship between perioperative changes in plasma arginine vasopressin and angiotensin Ⅱ and outcome in patients undergoing off-pump coronary artery bypass grafting / 中华麻醉学杂志

ObjectiveTo investigate the relatioaship between the changes in perioperative plasma vasopressin (VP) and angiotensin Ⅱ ( Ang Ⅱ ) concentrations and outcome in patients undergoing off-pump coronary attery bypass grafting (OPCABG).MethodsFifty ASA Ⅰ -Ⅲ patients (NYHA Ⅰ -Ⅲ ) of both sexes,aged 45-79yr,undergoing OPCABG,were enrolled in this study.Blood samples were collected before induction of anesthesia (T1,baseline),before skin incision (T2),at 10 and 30 min after skin incision (T3,T4 ),10 min after protamine injection (T5),end of operation (T6 ) and 24 h after operation (T7).Based on the intraoperative plasma VP concentrations,the patients were divided into high level group ( n =26) and low level group ( n =24) by hierarchical clustering analysis.The risk factors for perioperative lower plasma VP concentration were determined by logistic regression analysis.ResultsPlasma VP concentrations were significantly lower,while plasma Ang Ⅱ concentrations were significantly higher at T2-6 in the low level group than in the high level group.The incidence of vasoplegia (high cardiac output and low peripheral resistance) was significantly higher,the intra- and post-operative use of vasodilator was less,the tracheal extubation time,ICU stay and post-operative hospital stay were longer,and preoperative left ventricular ejection fraction (LVEF) was lower in low level group than in high level group.Logistic regression analysis showed that preoperative low LVEF was a risk factor for intraoperative low plasma VP concentration and OR was 1.122.Conclusion Plasma VP and Ang Ⅱ concentrations demonstrate an opposite trend of change during OPCABG.The incidence of vasoplegic syndrome is significantly higher and the outcome poor in low plasma VP group.Preoperative low LVEF is a risk factor for development of low plasma VP during OPCABG.

Primary culture and proliferation activity identification of rabbits subchondral bone cell / 国际生物医学工程杂志

ObjectiveTo study the method of cell isolation,primary culture and identification of subchondral bone cell of osteoarthritis(OA) rabbits.Methods The rabbit instable knee joint models were made by modified Hulth modeling method.The osteoblasts were harvested from the subchondral bone of rabbits by collagenase and tissue explants attachment.The morphology observation and biological identification were performed by inverted microscope and immunocytochemistry staining,respectively.The proliferative activity of cells were detected by MTT and the expression of Ⅰ-collagen at gene level was detected.ResultsThe cells started to appeared on the 11th day after the attachment.The cells form were fusiformis and triangle,the nucleolus were clear.The cultured cells had typical osteoblast morphological characteristics.The cells obtained from subchondral bone of rabbits were identified to be osteoblast by immunocytochemistry staining.The proliferative activity of cells were equably proliferation which detected by MTT.ConclusionThe modified method provides better way to obtain ideal subchondral osteoblast and the co-culture method is suitable for the study of OA microenvironment,which can simulate interactions of the subchondral osteoblast,synovial cells and chondrocyte.

Response of cyclic tensile strain on the human degenerative anulus fibrosus cells / 国际生物医学工程杂志

Objective To investigate the effects of different cyclic tensile strain on the proliferation of human anulus fibrosus cells from degenerated discs.Methods Anulus fibrosus(AF) cells were isolated from a degenerated human IVD,expanded in monolayer,and cyclically strained for 3 hours,applying 0,5％,10％,15％ and 20％ strains at a frequency of 0.25 Hz with the use of the DioDynamic test instrument.The flow cytometry method was used to examine the Af cells proliferation at 24 hours following application of the cyclic tensile strains.Results The proliferative index (PI) increased with the magnitude value of cyclic tensile strain except 20％ group.The most significant increase of proliferation index were found in 15％ group.Conclusion There might be some corelationships between magnitude of cyclic tensile strain and the proliferation of the degenerative AF cells.

In vitro culture and identification of synovial cell of rabbit knee osteoarthritis / 国际生物医学工程杂志

Objective To replicate experimental animal model of knee osteoarthritis and to investigate the method of culture and biological characteristics of rats synoviocytes of gonarthritis in vitro.MethodsAnimal models of knee osteoarthritis were made by the Modified Hulth method.4 weeks after the replicating experiment,synovial tissues were mechanically isolated and enzyme-digested and the growth of the synovial cells was investigated.Results The synovial tissues were obviously hyperplasia in the model made by the Modified Hulth method.The synovial cells were abundant after enzyme-digested cultivation and the cell activity was higher than 98%.Conclusion The study exhibits that the Modified Hulth method apparently promotes the hyperplasia of synovial tissues.The methods of isolation and cultivation of the synovial cells in vitro is proved to be simple and feasible.

Effect of antibody CR6261 V(H) and scFv expression on influenza virus infection / 生物工程学报

The emerging CR6261 antibody could neutralize several subtype influenza virus with high affinity, whose VH domain binds to the HA protein conserved domain. Therefore, it has drawn much attention as a potential broad-spectrum therapeutic antibody against influenza virus. In this study, we constructed the eukaryotic expression vectors pCR6261V(H), pCR6261V(H)-GFP and pCR6261scFv and screened the monoclonal cell lines that could stably express CR6261V(H), CR6261V(H)-GFP and CR6261scFv on the cell membrane. After influenza virus infecting the stable cell lines, the titers of viruses were tested by hemagglutination inhibition test. The result shows that the titers of viruses in CR6261scFv and CR6261V(H)-GFP stable expression cell lines decreased and there was no obvious discrimination between the CR6261V(H) expression cell line and the negative control, suggesting that CR6261V(H) and CR6261scFv expressing on the cell membrane could partly inhibit the virus infection. Though the effective inhibition strategy is undergoing, our research will provide new clues for the breeding of anti influenza transgenic animals.

A study on creating a promotion index system of medical quality in Grade Ⅲ hospitals in Beijing / 中华医院管理杂志

To explore how to create and optimize a promotion index system of medical quality evaluation, this article focuses on the hospital visiting process from patients, using analyzing collected those index system from couples of Grade Ⅲ hospitals in Beijing, and combining the results of literal study, field study and specialist consult, according to the different situation of general hospitals and specially hospitals, with the spirit of "maintaining the patients benefits, safeguarding the patients safety,and enhancing the medical quality", introduces the framework of the promotion index system, the rules to select the indicator, and so on, and discusses several problerns related to creating the index system.

Management practice to cope with A/H1N1 flu in the hospital / 中华医院管理杂志

r governments and mescal institutions in their prevention and control.

Cellular biocompatibility of shengji solution-gelatin-chitosan drug-loading skin scaffold / 中国组织工程研究

BACKGROUND: Tissue-engineered skin plays an important role in the repair and reconstruction of large-area skin lesion. Little is known about the effects of Chinese medicine-loading tissue-engineered skin scaffold on cell adhesion and wound surface infection. OBJECTIVE: To screen the application concentration of shengji solution, which can promote tissue regeneration, using keratinocytes and fibroblasts, and to observe the effects of shengji solution-gelatin-chitosan drug-loading skin scaffold on cellular adhesion and biocompatibility. DESIGN, TIME AND SETTING: Taking keratinocytes and fibroblasts as subjects, the present randomized and controlled experiment was performed at the Laboratory of Cell Engineering, Orthopedic Institute, Tianjin Hospital between February and August 2005. MATERIALS: One healthy big-ear rabbit was included for harvesting skin seed cells. Shengji solution, Danggui (Radix Angelicee Sinensis) and Shengdi (rehmannia dride rhizome) extract (self-extracted, 1.5 g/mL), gelatin-chitosan scaffold and shengji solution-gelatin-chitosan scaffold were provided by Tianjin University, China. METHODS: Passage 3 keratinocytes and fibroblasts were harvested by dispase Ⅱ- trypsase- ethylenediamine tetraacetic acid(EDTA) digestion. The prepared cells were divided into 5 groups: control, shengji solution, Danggui, Shengdi, and epidermal growth factor (EGF). Common culture solution containing 0.1 volume fraction of fetal bovine serum, shengjisolution (5, 6, 8, and 12 g/L), Danggui extract (8 g/L), Shengdi extract (8 g/L), and EGF culture medium (10 μ g/L) were used in corresponding groups for cell culture. At 1, 3, 5, and 7 days, cellular proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Keratinocytes and fibroblasts were cultured in different concentrations of shengji solution (50, 60, 80, and 120 g/L) and grouped according to different concentrations. At 7 and 14 days, cellular adhesion was observed by semi-quantitative method. At 4, 7, and 14 days, keratinocytes cultured by 60 and 80 g/L shengji solution were harvested for hematoxylin-eosin staining and subsequent scanning electron microscope observation. MAIN OUTCOME MEASURES: Effects of shengji solution on skin seed cell proliferation; effects of drug-loading scaffold on cellular adhesion; and cell-carrier biocompatibility. RESULTS: MIF detection results demonstrated that cells significantly proliferated after treatment of 5, 6, and 8 g/L shengji solution compared to the control group (P < 0.05). Obvious proliferation of passage 3 keratinocytes and fibroblasts was found on 60 and 80 g/L drug-loading scaffold. Keratinocytes on the drug-loading scaffold exhibited good cellular morphology and closely adhered to the scaffold. Shengji solution had no apparent toxic effects on cells. CONCLUSION: Shengji solution (60 and 80 g/L)-gelatin-chitosan scaffold can effectively promote the adhesion and proliferation of keratinocytes and fibroblasts.