Background The tissue-engineered cornea is becoming the hot spot in the ophthalmologic field,while the research of corneal substitute is in the ascendant,because it is more similar to the corneal morpha and easy to survive in vivo.Objective This study was to investigate the biocompatibility of recombinant human type-Ⅲ collagen/poly9 ( 3-( methacryloylamino ) propyl dimethyl ( 3-sulfopropyl ) ammonium hydroxide ) ( PMPDSAH ) interpenetrating polymer network (IPN) (RHC-Ⅲ/PMPDSAH IPN) hydrogel as a tissue-engineered cornea in rabbit eye and its feasibility as the corneal substitute.Methods One hundred and eight rabbits were randomly divided into experimental group( 90 rabbits) and normal control group ( 3 rabbits),and 15 rabbits ( 30 eyes ) used as the donor corneas.RHC-Ⅲ/PMPDSAH IPN,NGF PMPDSAH IPN and corneal grafts were lamellarly transplanted into the right eyes in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group respectively.The corneal transparency and neovascularization were examined and scored under the slim lamp and compared among three groups using Kraskal-Wallis H test.The corneal epithelization time was observed and compared among these three groups using one way analysis of variance and LSD-t test.The histological examination of corneas was performed at the 3rd day,1st and 2nd week,1 st,3rd and 6th month after the surgery.The immunohistochemistry was used to detect the expression of K3 in cornea at the 6th month.Results The grafts were well attached in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group,and no rejection reaction was found throughout 6-month following up.Compared with normal control group,no significant differences were found in the scores of corneal opacification and neovescularization in these three groups (x2 =4.34,P =0.23 ;x2 =2.60,P =0.46 ) at the 6th month.NGF PMPDSAH IPN group achieved reepithelialization in (4.97±0.63) days and was obviously shorted than that in RHC-Ⅲ/PMPDSAH IPN group and allograft group ( t =11.97,P =0.00; t =5.80,P =0.00).The re-epithelialization time in RHC-Ⅲ/PMPDSAH IPN was (6.86±0.71) days,and that of allograft group was (5.87±0.43 ) days,showing a significant difference ( t =6.32,P =0.00).Hematoxylin-eosin staining results demonstrated that implanted materials integrated into the host corneal tissue well and support corneal epithelialization.Part of the material degraded at the 2nd week and degraded completely 1 month later.Regular alignment and distribution of collagen fibers were seen in the regenerated cornea and were similar to those of the normal stroma in 6 months.Immunohistochemistry showed the positive expression of keratin-3 in corneal epithelial cells.Conclusions RHC-Ⅲ/PMPDSAH IPN has a good biocompatibility without toxicity to corneal tissue.Furthermore,NGF can promote the corneal wound-healing and re-epithelialization.The material can be used as safe and reliable corneal substitute after improving the mechanical strength.

Dendritic cells (DC) play an important roles in the maintenance of central immune tolerance and peripheral immune tolerance. DC can be involved in formation of autoimmune tolerance by many mechanisms and demonstrate strong plasticity, so that DC become hot issue in the research of transplantation tolerance recently. In this article the DC typing and its role, the indirect pathway of DC-inducing immune tolerance, the F1t3L and apoptotic cell role, the modified DC by genetic engineering and the immune inhibitors were summarized.
Dendritic Cells ; immunology ; Humans ; Immune Tolerance ; Transplantation Tolerance ; immunology

OBJECTIVES: Some changes are found in the labyrinth anatomy during postnatal development. Although the spatial orientation of semicircular canals was thought to be stable after birth, we investigated the age-related orientational changes of human semicircular canals during development. METHODS: We retrospectively studied the computed tomography (CT) images of both ears of 76 subjects ranged from 1 to 70 years old. They were divided into 4 groups: group A (1-6 years), group B (7-12 years), group C (13-18 years), and group D (>18 years). The anatomical landmarks of the inner ear structures were determined from CT images. Their coordinates were imported into MATLAB software for calculating the semicircular canals orientation, angles between semicircular canal planes and the jugular bulb (JB) position. Differences between age groups were analyzed using multivariate statistics. Relationships between variables were analyzed using Pearson analysis. RESULTS: The angle between the anterior semicircular canal plane and the coronal plane, and the angle between the horizontal semicircular canal plane and the coronal plane were smaller in group D than those in group A (P<0.05). The JB position, especially the anteroposterior position of right JB, correlated to the semicircular canals orientation (P<0.05). However, no statistically significant differences in the angles between ipsilateral canal planes among different age groups were found. CONCLUSION: The semicircular canals had tendencies to tilt anteriorly simultaneously as a whole with age. The JB position correlated to the spatial arrangement of semicircular canals, especially the right JB. Our calculation method helps detect developmental and pathological changes in vestibular anatomy.
Ear ; Ear, Inner ; Humans* ; Methods ; Parturition ; Retrospective Studies ; Semicircular Canals* ; Tomography, X-Ray Computed

Fractional Flow Reserve, Myocardial ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods

The study was aimed to detect the expressions of costimulatory molecules on CD3(+)CD4(+) T cells so as to accumulate informations for investigation of mechanism of myelodysplastic syndrome. 11 healthy blood donors as control and 38 patients with MDS de novo were studied. 38 MDS patients were divided into RA/RARS group and RAEB/RAEB-t group according to FAB classification. The expressions of CD28, CD154, CTLA-4, PD-1, CD25 on CD3(+)CD4(+) T cells in peripheral blood were detected by FCM. The results indicated that as compared with normal controls, the expression of CD28 in MDS patients decreased, and CD154 increased. The percentages of CTLA-4, PD-1 and CD25 in MDS were obviously higher than that in normal controls; the differences of CTLA-4, PD-1 and the ratio of CTLA-4/CD28 between RAEB/RAEB-t and RA/RARS were more significant with progressing of MDS. In conclusion, the expressions of costimulatory molecule in MDS patients were abnormal, which may be involved in the pathogenesis of MDS.
Adult ; Antigens, CD ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; CD28 Antigens ; metabolism ; CD40 Ligand ; metabolism ; CTLA-4 Antigen ; Case-Control Studies ; Female ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Lymphocyte Count ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; Programmed Cell Death 1 Receptor ; T-Lymphocytes ; immunology ; metabolism ; Young Adult

OBJECTIVETo Verify the safety and reliability of one-stage repair of complete cleft Lip and palate in infancy and to obtain the primary result.

METHODSThe simultaneous repair of complete cleft Lip and palate in infants 3 to 12 months of age were performed in 271 cases. The deformities include 185 cases of typical complete unilateral clefts and 75 cases of complete bilateral clefts, and other 11 atypical cleft infants. The preoperative orthopedic treatment for wide alveolar cleft was undertaken in 24 infants and the lip appearance and speech outcome were evaluated in 116 children by 1 to 4 years' postoperative follow-up.

RESULTSAll infants, except for dyspnea in 2 babies, palatal fistula formation in 6 cases and temporary wound hemorrhage in 5 infants, were recovered without complications. After orthopedic treatment, the width of the alveolar cleft was reduced 6.1 mm in average. The evaluation showed that 93.1% of children had got good or excellent lip appearance. And the acceptable or excellent speech was found in 94.8% children.

CONCLUSIONSSimultaneous repair of complete cleft lip and palate in infancy is safety and reliable. The preoperative orthopedic procedure is able to reduce the wide alveolar cleft and to achieve alignment of alveolar segments. The acceptable and or excellent lip appearance and speech function could be obtained in this one-stage operative procedure in infants.

Cleft Lip ; surgery ; Cleft Palate ; surgery ; Female ; Humans ; Infant ; Male ; Treatment Outcome

OBJECTIVETo observe the long-term effect of tissue engineering-based repair of large weight-bearing bone defect in goats, and the final outcome of the scaffold material coral hydroxyapatite (CHAP) in vivo.

METHODSFifteen Chinese goats were subjected to operations to induce a 2-cm left tibial diaphyseal defect, which was filled subsequently with CHAP and bone marrow stromal stem cells (BMSCs). The repaired defects were evaluated by ECT, X-ray and histology in the early stage and at 6, 12, 18, and 24 months postoperatively.

RESULTSECT showed good bone regeneration and revascularization within 2 months postoperatively. X-ray and histology displayed eccentric and gradual bone regeneration in the early stage, and the tissue-engineered bone graft was firmly healed with the goat tibia. X-ray and histological examination at 6, 12, 18, 24 months postoperatively revealed moulding of the new bones and medullary cavity recanalization, and the structure of CHAP disappeared and gradually integrated into the new bones.

CONCLUSIONTissue-engineered bone is capable of total repair of large bone defect in goats by forming normal functional new bones. CHAP can be eventually degraded completely and become the component of the newly generated bones.

Animals ; Bone Marrow Transplantation ; Bone Regeneration ; Bone Substitutes ; Cells, Cultured ; Goats ; Prostheses and Implants ; Tibial Fractures ; physiopathology ; surgery ; Tissue Engineering ; methods

OBJECTIVETo investigate the role of PERK/eIF2alpha signaling pathway in hepatocyte apoptosis of alcoholic liver injury rats.

METHODSRat models with ethanol-induced liver injury were successfully developed by gastric gavage with ethanol-corn oil mixtures for 12 weeks. At different time points (4, 6, 10, 12 week), liver pathology was dynamically observed. The hepatocyte apoptosis was quantitatively analyzed by Annexin V-FITC/PI double-labeled flow cytometry, the serum total homocysteine (tHCY) level was detected by ELISA and the expressions of eIF2a, p-eIF2a, GRP78/Bip, GRP94, caspase-3 and caspase-12 in liver were examined using Real-time PCR and Western blot.

RESULTSTypical acute liver injury and chronic liver injury were observed at week 4 and week 12 respectively. The hepatocyte apoptosis rates in 6-week model rats significantly increased compared with normal rats (P value less than 0.05), and the degree of hepatocyte apoptosis continued to increase with the modeling time, and the percentages of early and total apoptosis reached 26% and 29% at week 12. From week 6 to week 12, the serum tHCY levels in model rats were obviously higher than in normal rats (P value less than 0.01). Since week 4, eIF2a protein phosphorylation in model rat livers remarkably elevated compared with that in normal rat livers (P value less than 0.01), and at week 12 the peIF2a protein expression in model rat livers increased by 2.81-fold. Since week 4 the expressions of GRP78/Bip, GRP94, caspase-12 and caspase-3 mRNA and protein in model rat livers showed a significant increase as compared to normal rat livers, and at week 12, these gene and protein levels increased 4.70, 12.95, 3.83, 4.05 fold and 3.93, 6.93, 9.88, 3.31 fold, respectively (P value less than 0.01).

CONCLUSIONActivation of PERK/eIF2a signaling pathway contributes to the occurrence and development of hepatocyte apoptosis in alcoholic liver injury rats and it might be as a potential target for therapeutic applications in alcoholic liver diseases.

Animals ; Apoptosis ; Eukaryotic Initiation Factor-2 ; metabolism ; Hepatocytes ; cytology ; pathology ; Liver Diseases, Alcoholic ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; eIF-2 Kinase ; metabolism

This study was aimed to investigate the effect of 5-azacytidine (5-AZA) on XAF1 expression in myeloma cells and efficacy of 5-AZA treatment for myeloma in vitro. XAF1 expression was analyzed by semi-quantitative PCR. Methylation-specific PCR (MSP) was used to detect the methylation status of XAF1 promoter CpG islands. RPMI 8226 and XG-7 cells were treated with 0-5 micromol/L of 5-AZA. Expression of XAF1 mRNA variants was confirmed by gel electrophoresis. The results indicated that the untreated RPMI 8226 cell expressed XAF1 mRNA transcript 1 and transcript 2, untreated XG-7 cells did not express XAF1 mRNA. Hypermethylation of XAF1 promoter CpG islands could be detected in both cell lines. Both cell lines expressed full-length XAF1 transcript after being treated with 2.5 micromol/L of 5-AZA for 72 hours. 5-AZA treatment led XAF1 promoter CpG island to hypomethylation in both cell lines. 5-AZA exerted anti-myeloma activity in a time- and concentration-dependent manner. The IC(50) value of XG-7 cells treated with 5-AZA for 48 hours was 2.6 micromol/L. 1.0, 2.0, 2.5 and 5.0 micromol/L of 5-AZA treatment for 48 hours induced (34.3 +/- 8.0)%, (54.8 +/- 3.1)%, (64.1 +/- 3.4)%, (81.0 +/- 4.1)% apoptosis in XG-7 cell line respectively. The combination of 1.0 - 4.0 micromol/L of 5-AZA with 1.0 - 4.0 micromol/L of arsenic trioxide (ATO) exhibited synergistic toxicity in myeloma cells with all CI values less than 1.0. It is concluded that lack of XAF1 expression and abnormal expression of XAF1 in myeloma cell lines are associated with the hypermethylation of XAF1 gene promoter CpG island. 5-AZA treatment can induce the expression of XAF1 mRNA and protein in myeloma. 5-AZA exerts anti-myeloma activity via apoptosis at clinically achievable concentrations. The findings suggested that 5-AZA and ATO may be an effective combination in the therapy of patients with multiple myeloma.
Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Azacitidine ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Multiple Myeloma ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic

OBJECTIVETo compare the characteristics of immunomagnetic beads and hespan precipitation for isolation of mononuclear cells (MNCs) from umbilical cord blood and try to find a better isolation method for MNCs.

METHODSFifteen umbilical cord blood samples from healthy parturiens were collected between December 2007 and March 2008. MNCs were isolated using hespan precipitation and CD133 immunomagnetic beads, respectively. MNCs were identified using the surface marker CD34 by flow cytometry on the 30th of primary culture. Growth conditions and morphologic changes of primary cells were observed by an inverted microscope.

RESULTSThe number of MNCs from umbilical cord blood isolated by hespan precipitation (15.23 +/- 4.30 x 10(6)/mL) was significantly greater than that by CD133 immunomagnetic beads (0.066 +/- 0.027 x 10(6)/mL) (p<0.05). The MNCs isolated by hespan precipitation suspended at the culture medium and their growth was slow after passage. The growth of MNCs isolated by CD133 immunomagnetic beads was kept in a good condition. The CD34 positive rate of MNCs isolated by hespan precipitation and immunomagnetic beads was 10.1% and 0.5%, respectively.

CONCLUSIONSThe hespan precipitation is an effective method for MNCs isolation from human umbilical cord blood, but with a cell growth condition below the mark. The MNCs isolated by CD133 immunomagnetic beads are in a high purity quotient.

AC133 Antigen ; Antigens, CD ; immunology ; Antigens, CD34 ; analysis ; Cell Separation ; methods ; Chemical Precipitation ; Fetal Blood ; cytology ; Glycoproteins ; immunology ; Humans ; Hydroxyethyl Starch Derivatives ; chemistry ; Immunomagnetic Separation ; methods ; Infant, Newborn ; Leukocytes, Mononuclear ; cytology ; Peptides ; immunology