Child ; Critical Care ; Humans ; Intensive Care Units, Pediatric ; Medical Records ; Severity of Illness Index

The source of seed cells has always been the major problem in cartilage tissue engineering.Bone marrow stromal cells (BMSCs) have gradually become an optimal source of seed cells for cartilage engineering due to their high proliferative potential,multi-lineage differentiation potential and easiness to be obtained with minute trauma.The great challenge is how to get abundant BMSCs with a high purity and how to induce them in vitro into chondrogenic phenotype.This review aims to discuss the various strategies that can induce BMSCs chondrogenic differentiation in vitro so as to offer beneficial reference for constructing cartilage with BMSCs as seed cells.

Background Vitronectin is a glycoprotein that has a variety of functions.Its expression was markedly higher in the retina of oxygen induced mice,which was confirmed in our animal model,and also increased in human umbilical vein endothelial cells (人 UVECs) that were cultured in high glucose.However,there was no evidence that showed vitronectin was involved in retinal neovascularization.Objective This study was to observe the influence of vitronectin on cytoskeleton remodeling,cell migration and blood vessel formation in 人 UVECs conditioned by high glucose.Methods 人 UVECs were cultured in high glucose and the expression of vitronectin was knocked down using RNA interference technology.The experiments were divided into the high glucose group (人 UVECs were conditioned with DMEM medium that contained 50 mmol/L glucose),negative interference group (人 UVECs were transfected with control siRNA in advance,and then were conditioned with DMEM medium that contained 50 mmol/L glucose) and positive interference group (HUVEC were transfected with vitronectin siRNA in advance,and then were conditioned with DMEM medium that contained 50 mmol/L glucose).The protein expression of vitronectin was measured by Western blot,and the microfilament cytoskeleton of 人 UVECs was examined by immunofluorescence cytochemical staining followed by fluorescence microscopy.Cell migration ability in a scratch wound assay and blood vessel formation ability in a matrigel assay of 人 UVECs were evaluated.The general differences were analysed by One-Way ANOVA ;further contrasts of the two groups were analysed by the LSD-t test.Results The differences in vitronectin expression of the three groups were not obvious at 0 hour (F=1.064,P＞0.05).After 24 hours,vitronectin expression was highest in the high glucose group,lower in the negative interference group,and the lowest in the positive interference group,and the differences were significant (F =15.519,P＜0.05).After 48 hours,vitronectin expression of the three groups displayed the same pattern,and the differences were also significant (F=37.521,P＜0.05).Immunofluorescence showed that the cytoskeleton structure was most obvious in the high glucose group,moderate in the negative interference group,and was the least obvious in the positive interference group,after both 24 hours and 48 hours.In the scratch wound assay,the cell migration ability of the high glucose group was the highest,lower in the negative interference group,and the lowest in the positive interference group after 24 hours,and the differences were significant (F=90.685,P＜0.05).After 48 hours,the cell migration abilities of the three groups displayed the same pattern,and the differences were also significant (F=67.880,P＜0.05).In the matrigel assay,after 6 hours,the number of blood vessels formed in the high glucose group was more than that in the negative interference group,and the least amount was found in the positive interference group.The differences among of them were significant (F =86.653,P＜0.05).The number of blood vessel formed in the positive interference group was also the lowest after 12 hours,and the differences were also significant (F=18.992,P＜0.05).Conclusions Vitronectin can bring about cytoskeleton remodeling,increase in cell migration,and enhancement of blood vessel formation in 人 UVECs conditioned in high glucose.It may be one of the important influence factors of diabetic retinopathy.

OBJECTIVE: To study the objectivity and reliability of needle electromyography and nerve conduction for detection of musculus extensor digitorum brevis strength, which may provide a basis for establishing a quantitative detection of muscle strength in forensic clinical study. METHODS: Forty-four healthy people were enrolled as the subjects, and during toe dorsiflexion, the following items including needle electromyography indexes, motor unit potential (MUP) amplitude, MUP count, recruitment reaction type, and nerve conduction detection indexes, compound muscle action potential (CMAP) amplitude, CMAP latent period and motor nerve conduction velocity (MNCV), were simultaneously detected under the cooperation and disguise condition. RESULTS: Under the cooperation condition, regardless of the same operator or different operators, there were good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, while there were normal test-retest reliabilities in MUP count and recruitment reaction type and the repeatability of the same operator was slightly better than the repeatability between different operators. Under the disguise condition, test-retest reliabilities of MUP amplitude, CMAP amplitude, CMAP latent period and MNCV were relatively high, while test-retest reliabilities of MUP count and recruitment reaction type were relatively low. CONCLUSION There are good test-retest reliabilities in MUP amplitude, CMAP amplitude, CMAP latent period and MNCV, which can be conducive to comparison between different operators and results at various times; MUP count and recruitment reaction type, which can be easily affected by subjectivity of operators and examinees, can be used to differentiate whether an examinee disguises or not. The indexes used to objectively judge muscle strength remain to be further investigated.
Electrodes, Implanted ; Electromyography ; Humans ; Muscle Strength/physiology* ; Muscle, Skeletal/innervation* ; Neural Conduction/physiology* ; Reproducibility of Results ; Toes

OBJECTIVETo study the role of the soluble factors secreted by tissue engineered cartilage in promoting bone marrow stromal cells (BMSCs) chondrogenesis as an important aspect.

METHODSPorcine BMSCs, chondrocytes and dermal fibroblasts were respectively in vitro expanded and then seeded onto the polyglycolic acid/polylactic acid (PGA/PLA) scaffold. After 3 days, they were indirectly co-cultured by transwell. BMSCs-scaffold constructs were co-cultured with chondrocytes-scaffold constructs as experiment group (Exp), while co-cultured with fibroblasts-scaffold constructs as control group. BMSCs with the same cell number were seeded onto the scaffolds as another control group. There were 3 specimens in each group. All specimens were harvested after in vitro indirect co-culture for 8 weeks. Gross observation, histology, immunohistochemistry and RT-PCR were used to evaluate the results.

RESULTSThe BMSCs-scaffold constructs co-cultured with chondrocytes-scaffold shrunk gradually during in vitro culture, but formed the mature lacuna structures and metachromatic matrices, collagen II expression could be observed by immunohistochemistry and RT-PCR examination. In the control group, the constructs shrunk greatly during in vitro culture and showed mainly fibrous tissue.

CONCLUSIONSThe soluble factors secreted by chondrocytes can solely induce chondrogenic differentiation of BMSCs and thus promote the in vitro chondrogenesis of BMSCs.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; secretion ; Chondrogenesis ; Coculture Techniques ; Female ; Male ; Stromal Cells ; cytology ; Swine ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo observe the effect of Pinggan Qianyang Recipe (PQR) on inhibiting angiotensin II (Ang II) induced proliferation and migration of vascular smooth muscle cells (VSMCs) and changes of DNA methylation.

METHODSVSMCs were cultured using tissue explant method, and PQR containing serum was prepared. Primarily cultured VSMCs were divided into four groups, the normal group, the model group, the folate group (folic acid intervention) , and the PQR group. The proliferation and migration of VSMCs was duplicated by Ang II. After 24-h Ang II induced culture, 40 microg/mL folic acid was added to the folate group for 48 h, while 5% PQR containing serum was added to the PQR group for 48 h. The cell growth curve of VSMCs was drawn by using Cell Counting Kit (CCK-8). The proliferative activity of VSMC was determined by MTT assay. The migration of VSMCs was measured by Millicell chamber. The general level of cytosine methylation in cell nucleus was detected via 5-mC antibodies immunofluorescence, and mRNA expression levels of DNA methyltransferase 1 (DNMT1) were measured by Real-time q-polymerase chain reaction (q-PCR).

RESULTSVSMCs were promoted by Ang II at 10(-6) mol/L for 24 h. Compared with the normal group, the proliferative activity and migration quantity of VSMCs obviously increased, and DNA methylation level obviously decreased (P < 0.05, P < 0.01). Compared with the model group, the cell growth, proliferative activity and migration quantity of VSMCs obviously decreased and the general DNA methylation level increased in the folate group and the PQR group (P < 0.05, P < 0.01). Compared with the normal group, the mRNA expression of DNMT1 decreased in the model group (P < 0.01). Compared with the model group, mRNA expression of DNMT1 in Ang II induced VSMCs was obviously enhanced in the folate group and the PQR group (P < 0.01).

CONCLUSIONSPQR could inhibit Ang II induced proliferation and migration of VSMCs, and cause high genomic DNA methylation level. Changes of DNA methylation might be associated with DNMT1 expression.

Angiotensin II ; pharmacology ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects

OBJECTIVETo explore the feasibility of constructing tissue-engineered cartilage with human dermal fibroblasts (HDFs) in vitro.

METHODSPorcine articular chondrocytes and HDFs were isolated and in vitro expanded respectively. Then they were mixed at the ratio of 1:1 (chondrocytes: fibroblasts) . The mixed cells were seeded onto polyglycolic acid (PGA) scaffold at the ultimate concentration of 5.0 x 10(7)/ml as co-culture group. Chondrocytes and HDFs at the same ultimate concentration were seeded respectively onto the scaffold as chondrocyte group ( positive control group) and fibroblast group ( negative control group). The specimens were collected after in vitro culture for 8 weeks. Gross observation, histology and immunohistochemistry were used to evaluate the results.

RESULTSIn chondrocyte group, the cell-scaffold constructs could maintain the original size and shape during in vitro culture. The new formed cartilage-like tissue had typical histological structure and extracellular matrix staining similar to normal cartilage. In co-culture group the constructs shrunk slightly at 8 weeks, cartilage-like tissue formed and GAG could be detected for strong expression by Safranin O staining. Furthermore, using the specific identification, a few HDFs derived cells were found to form lacuna structure at the peripheral area of cartilage-like tissue. In fibroblast group, the constructs deformed and shrunk gradually without mature cartilage lacuna in histology.

CONCLUSIONThe 3D-co-culture system can effectively induce the differentiation of HDFs to chondrocytes. The tissue-engineered cartilage can be constructed in vitro with the 3D-co-culture system.

Animals ; Cartilage ; cytology ; Cells, Cultured ; Chondrocytes ; cytology ; Coculture Techniques ; Dermis ; cytology ; Fibroblasts ; cytology ; Humans ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective To explore the detection methods for cognitive dysfunction of the major depression in Elderly and analyze their clinical significance.Methods Using matched-pairs study,42 patients with seniie de- pressive disorders(experimental group)and 42 normal aged people(control group)were examined with auditory e- voked potential P300(event related potential,ERP-P300)and SECF,respectively.Results It was found that the scores with registration,span,recall,classification and total score of the subjects in the experimental group were sig- nificantly lower than those in the control group(P

OBJECTIVETo test the hypothesis that tissue-engineered cartilage can be bioincorporated with a nonreactive, permanent endoskeletal scaffold.

METHODSChondrocytes obtained from swine articular were seeded onto polyglycolic acids(PGA) scaffold which was incorporated with high-density polyethylene (Medpor). After cultured in vitro for two weeks,the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. Six weeks later,the newly formed cartilage prosthesis was harvested, and a small part of sample was evaluated by gross view, histology, type II collagen immunohistochemistry and biochemistry. PGA scaffold seeded with cells as the control group.

RESULTSThe newly formed cartilage was very similar to normal cartilage in both gross view and histology, and jointed Medpor tightly. The center of control group was hollow.

CONCLUSIONThis pilot technique combining tissue engineering with a permanent success in creating cartilage without "hollow" phenomenon. biocompatible endoskeleton demonstrated

Animals ; Biocompatible Materials ; Cartilage ; transplantation ; Chondrocytes ; cytology ; Materials Testing ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Pilot Projects ; Polyethylenes ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo explore the relationships between quality of life, negative life events, social support and suicide ideation among undergraduates in colleges.

METHODS3517 undergraduates in colleges were recruited by multistage stratified random clustered sampling method. Factors associated with suicide ideation were analyzed with logistic regression by scores of Beck Scale for Suicide Ideation(BSSI), Generic Quality of Life Inventory (GQOLI), Adolescent Self-rate Life Events Checklist (ASLEC), Social Support Rating Scale (SSRS) and a questionnaire on background information.

RESULTSThe rate of suicide ideation within 7 days was 14.1%, especially in females (15.96%), with single parent (23.79%) and disabled undergraduates (25.00%). The primary risk factors for suicide ideation were with low psychological function, material life, family/social support, lower availability of support and more negative life events.

CONCLUSIONThe prevalence of suicide ideation among these undergraduates was high, appropriate measures focusing on these risk factors should be implemented.

China ; epidemiology ; Cluster Analysis ; Female ; Humans ; Logistic Models ; Male ; Risk Factors ; Self-Injurious Behavior ; epidemiology ; Students ; psychology ; Suicide ; psychology ; Surveys and Questionnaires