Objective To investigate the histopathologic characteristic of the vitreous herniation out of sclerotomy site during vitrectomy. Methods Twenty specimens of tissues herniated at vitrectomy site were collected. The paraffin sections or fresh smears were stained with hematoxylineosin and examined under light microscope. The specimens were collected from the affected eyes with rhegmatogenous retinal detachment (9 cases), traumatic retinal detachment (1 case), miscellaneous vitreous hemorrhage (6 cases) and intraocular foreign body (4 cases). Results The herniated tissues were found to be retina in 4 cases, ciliary tissue in 1 case, retina and ciliary tissue in 1 case, uvea in 1 case,and hyaloid tissue in 13 cases. Conclusion There were not only vitreous, ciliary epithelial cells and pigment contained epithelia, but also ciliary body, retina and uvea in the prolapsed tissues of sclerotomy site, which might be related to the occurence of some clinical complications.

Objective To determine the expression of the growth factors and the receptors related to angiogenesis in the intraocular tissues incarcerating in the sclerotomy sites. Methods Ten specimens from prolapsing intraocular tissues in sclerotomy sites during vitrectomy were obtained and serially sectioned in cryostate and were stained with a group of polyclonal antibodies against vascular endothelial growth factor(VEGF), basic fibroblast growth factor (bFGF), platelet derived growth factor A(PDGF A) and transforming growth factor ? 1(TGF ? 1) as well as their receptors by using a streptavidin peroxidase system. Results The tissues prolapsed from the sclerotomy sites were identified as retina(3 cases), vitreous tissues(3 cases), degenerated red blood cell components(2 cases), ciliary body(one case) and fibrous tissue(one case). All specimens expressed VEGF and bFGF as well as their receptors. PDGF A, TGF ?1 and their receptors expressed in the most of specimens. The positive cells included retinal cells, ciliary non pigmented epithelial cells and pigmented epithelial cells, fibrous cells and the cells in vitreous. Conclusions The intraocular tissues incarcerated in the sclerotomy entries express the growth factors and receptors related to angiogenesis. This might be one of the potential factors of developing anterior proliferative vitreoretinopathy.

BACKGROUND: Embryonic stem cells (ESCs), the seed cells of all mature cells in vivo, are useful tools for nerve transplantation and developmental gene function research. Notch1 signaling pathway is the key pathway to control the ordered neural development and differentiation of many kinds of neural cells, however, there is no report on the dynamic expression of Notch1 signal during the ESC differentiation to date. OBJECTIVE: To investigate the expression of Notch1 protein, transmembrane signal transduction molecule, during directional differentiation of embryonic stem cells into neural cells. DESIGN, TIME AND SETTING: Cell research was carried out between October 2003 and October 2004 at Zhongshan Ophthalmic Center, SUN Yat-sen University, Guangzhou, Guangdong Province, China. MATERIALS: BALB/C mouse embryonic stem cell line Ⅵ (passage 11)was obtained from experimental animal center of SUN Yat-sen University, provided by professor Huang Bing. ESC culture medium was high-glucose DMEM medium with 20% bovine serum and 106 IU/L mouse leukemia inhibitory factor. Induced differentiation medium was high-glucose DMEM medium with 20% fetal bovine.serum and 5×107 mol/L retinoid acid(RA). METHODS: Passage 11 ESCs were resuscitated and incubated by ESC culture medium in incubator at 37℃ with 5% CO2. Passage 11 ESCs were subcultured after 2 or 3 days and RA was added into medium to induce differentiation. Three time points for observation were established: induced for 1, 5 and 9 days. MAIN OUTCOME MEASURES: Morphological changes were observed under inverted phase contrast microscope, MAP-2 antigen expressed in differentiated cells was detected by immunofluorescence method. Immunocytochemistry, Western Blot, flow cytometry assay were used to investigate the Notch1 protein expression. RESULTS: ESCs presented clone-like growth. After induced by RA for 9 days, single neural network was achieved around most of the cell clusters. With the prolongation of induction, MAP-2 positive neural cells increased gradually. Almost all ESC clones expressed Notch1 protein strongly or positively, but Notehl protein expression decreased gradually after induced differentiation (P < 0.01). CONCLUSION: Notch1 signal shuts off progressively during induction of ESCs into neural cells, which suggests Notch1 may play an important role in the differentiation of ESCs into neural cells.

BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.

Objective To investigate the correlation of serum total bile acid (TBA) levels with the inflammation grades of liver tissue in chronic liver diseases.Methods Cyclophorase assay was used to detect the serum TBA levels in 172 patients with various chronic liver diseases,and the inflammation grades of liver tissue were determined by liver biopsy.The correlation between serum TBA levels and the inflammation grades of liver tissue was evaluated using SPSS 12.0 software.Results Serum TBA level was positively correlated with the inflammation grade of liver tissue ( r =0.275,P ＜ 0.01 ).The inflammation grade reached G2 when serum TBA was 20 μmol/L.Conclusion Serum TBA level may be useful for evaluating the inflammation grade of liver tissue in chronic liver diseases.

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.

Objective:To study effects of large dose thymopeptides on T cell subpopulations of patients with malignant tumor under radiotherapy.Methods:Fifty one patients with malignant tumor under radiotherapy were divided into 2 groups with 100 mg and 200 mg thymopeptides respectively.The patients we re given thymopeptides,100 mg/d or 200 mg/d,iv, for 10 days.The positive percent ages of CD4,CD8,CD25 and CD56 in T cells of peripheral blood before and after thymopeptide treatment were determined by flow cytometry.Results:The positive percentages of CD4 and CD25 in T cells of peripheral bl ood after 100 mg/d thymopeptide treatment were significantly higher than those befor e thymopeptide treatment (P<0.05),while those of CD4，CD8，CD25 and CD56 in T cells of peripheral blood after 200 mg/d thymopeptide treatment all increased significantly (P<0.05 or P<0.01). Conclusion:These results suggest that large dose of thymopeptides can increase i mmune function of patients with malignant tumor under radiotherapy,and the curat ive effect of 200 mg/d thymopeptides is better.

AIM: To investigate the feasibility and effect of directly differentiation of embryonic stem cells(ESC) into neural cells induced by retinoid acid(RA) without embryonic body(EB) culture period in vitro.METHODS: ESC were digested and divided into 4 groups: group A and B were undergone EB culturing.After that,cells in group A were induced by RA,cells in group B were differentiated spontaneously,cells in group C were committedly induced by RA directly without EB culturing,and cells in group D were differentiated spontaneously without EB period.Morphologic changes were observed under inverted microscope and scanning electron microscope.MAP-2 and GFAP were detected by immunocytochemistry and flow cytometry after differentiated for 9 days.RESULTS: In groups A or C,neuron-like cells increased gradually,forming neural network.At the 9th day,a large part of cells in these groups were MAP-2 positive cells,and the positive rate was higher than that in groups B or D(P0.05).CONCLUSION: ESC was directly induced into neural cells by RA without EB culture period in vitro.This modified method has the same effect as the traditional RA 4-/4+ assay and can replace the traditional method.

OBJECTIVETo explore the effect of the cytoplasmic DNA sensor DAI on replication of hepatitis B virus (HBV) and its possible mechanism.

METHODSThe hepatocyte-derived cell line HepG2 was co-transfected with DAI siRNA and the HBV1.3 replicative plasmid PHY106, and the cells were divided into two experimental groups. Six hours later, total RNA was extracted from the first group of cells and expression of IFIT1 and IL-6 were detected by real-time RT-PCR. The second group of cells was incubated for 4 days, after which the cell supernatant was collected and the HBV surface antigen (HBsAg) and envelope antigen (HBeAg) were detected by ELISA. In addition, HBV core particles were extracted and applied to southern blot assay to detect the intracellular HBV replication intermediates (rcDNA, dlDNA and ssDNA). Next, the HepG2 cells were triple transfected with siRNA targeting the type I interferon pathway molecule TBK1 and DAI simultaneously and HBV1.3, after which HBV viral proteins were detected. Two-group comparisons were made using the independent sample t-test, and more-than-2-group comparisons were made using ANOVA.

RESULTSDAI gene expression was down-regulated in response to DAI siRNA transfection. Cells with down-regulated DAI showed inhibited HBV replication (in a dose-dependent manner), accompanied by reduced levels of HBsAg (0.0195+/-0.0050 vs.

CONTROL0.3150+/-0.0200, P less than 0.05, t = 14.77) and HBeAg (0.0140+/-0.0040 vs.

CONTROL0.01235+/-0.0135, P less than 0.05, t = 7.777). No effect of down-regulated DAI was observed for the expression of IFIT1 of IL-6. siRNA-mediated down-regulation of TBK1 and DAI simultaneously led to reduced expression of HBsAg and HBeAg.

CONCLUSIONDown-regulation of DAI gene expression inhibited HBV replication and HBV protein expression, but the underlying mechanism was not related to the type I interferon or NF-kB signaling pathway.

Carrier Proteins ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Down-Regulation ; Gene Expression Regulation ; Hep G2 Cells ; Hepatitis B Surface Antigens ; isolation & purification ; Hepatitis B e Antigens ; isolation & purification ; Hepatitis B virus ; physiology ; Humans ; Interleukin-6 ; metabolism ; NF-kappa B ; metabolism ; Plasmids ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transfection ; Virus Replication

Objective: To define radiographic features that response to serial casting and bracing for progressive early-onset scoliosis (EOS). Methods: A retrospective study of a total of 20 patients (10 females and 10 males) with complete radiographic data diagnosed as progressive early onset scoliosis treated with serial cast or brace for at least 12 months in the 306th Hospital of PLA from June 2011 to April 2018. Ages at initial diagnosis were all less than 5 years old. They were divided into two groups according to the main curve degree, those with cobbs angles more than 50 degree treated with serial cast, or else with brace. All the cases have radiographs of pretreatment, posttreatment, and last follow-up, and anteroposterior (lateral) film of the full length spine in standing position were taken to evaluate magnitudes and balance of coronal and sagittal malformations. We compared the general data of the two groups by independent sample t test and that of pretreatment, posttreatment and the last follow-up by paired-sample t test. Results: According to the effective standard of not less than 10 degrees improvement, 9 of them were effective, 4 cast and 5 brace, the effective rates were 44.44% and 45.45%, 3 cases from brace group progressed. The magnitudes of the main curve improved significantly after the first treatment of cast and brace (55.4±24.36 vs 42.35±23.62) degree (t=5.850, P=0.000), and at the last follow up, the curve decreased slightly compared of that before interventions (55.4±24.36 vs 51.8±26.33). The compensatory curves, segmental kyphosis, thoracic kyphosis, lumbar lordosis, balance of both coronal and sagittal also had no significant differences between the primal and the latest follow up. The widths 159.63±19.27 mm, 160.81±14.54 mm, 176.08±28.10 mm (t=3.942, P=0.001; t=-3.096, P=0.006) and heights 153.78±29.24 mm, 161.14±29.53 mm, 175.01±36.91 mm of the thoracic were significantly different between pre-posttreatment and the last follow up (t=-5.950, P=0.000; t=-3.997, P=0.001). Serial cast and brace application can preserve the growth of the thorax. Conclusion Serial cast and brace are viable growth friendly methods to deal with progressed EOS, Although a cure improvement cannot be always expected, they can stabilize relatively large curves in young children and may help delay eventual surgical intervention.