Background Light-induced retinal damage results in the damage of retinl pigment epithelial (RPE) cells and therefore affects the pathogenesis and development of age-related macular degeneration (AMD).Studies showed that tissue factor (TF) is overexpressed in oxidative damaged RPE cells and the choroidal neovascularization (CNV) of AMD,speculating that the suppression of TF can prevent the damage of RPE cells and inhibit CNV.Objective This study was conducted to observe the protective effects of TF targeting peptide (TFTP),a new drug of autologous synthesis,on human RPE-cells induced by blue light.Methods Human RPE cells were isolated from donor eye and cultured.Cultured cells were divided into blank control group,model group and TFTP treated group.Light-induced RPE cell damage model was established by exposuring the cells in the blue light of (4.0±-0.5) mW/cm2 for 12 hours in the model group,and different concentrations (10,100,150,200,300 μmol/L) of TF-TP were added into the medium to pretreat the cells for 24 hours and then exposed the cells to the blue light for 12 hours in the TF-TP groups.The cell viability was determined by CCK-8 assay.The morphology and ultrastructure in the cells were observed under the inverted microscope and transmission electron microscope.The apoptosis of the cells was assayed by Hoechst staining.The expressions of TF and apoptosis-related protein bax,bcl-2 in the cells were determined by Western blot.Results CCK-8 assay showed that there was no significant difference in the cell viability among blank control group and different concentrations TF-TP groups (F=2.15,P =0.11).The cell survival rate of blank control group,model group and 150 μmol/L TF-TP group was (100.0±0.00) ％,(43.79±6.55) ％ and (63.45±3.57) ％,and the survial rate was increased in the 150 μmol/L TF-TP group compared with the model group (P =0.00),and 150 μmol/L was detemined as a optimal concentration of TF-TP.A lot of shrinkage,deformation,suspension cells were exhibited under the optical microscope,and decrease of microvilli structure,rupture of mitochondrial cristae and vacuolar degeneration of the cells were found in the model group,and the damage of the cells were evidently lightened in the 150 μ mol/L TF-TP group.The apoptosis rate of the cells were (0.98 ±0.19)％,(9.98 ±0.82) ％ and (5.73 ±0.88) ％ in the blank group,model group and 150 μmol/L TF-TP group,respectively,with a significant difference among the groups (F =206.18,P =0.00),and the apoptosis rate of the cells in the 150 μmol/L TF-TP group was significantly lower than that in the model group (P＜0.05).Compared with the blank control group,the relative expression of bax and TF was obviously increased and that of bcl-2 was decreased in the model group;while the expression of bax and TF was lower,and that of bcl-2 was higher in the 150 μmol/L TF-TP group compared with the model group (all at P ＜ 0.05).Conclusions Pretreation of TF-TP can lessen cell apoptosis and increase cell survival rate and therefore plays a protective role to blue light-induced human RPE cells possibly by inhibiting bax/bcl-2 apoptotic pathways mediated by TF.

Calbindin 2 ; metabolism ; Heart Neoplasms ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Rhabdomyosarcoma, Embryonal ; metabolism ; pathology ; surgery

Objective To investigate the therapeutic effect and side effects of concurrent chemotherapy and intensity-modulated radiotherapy (IMRT) following induction chemotherapy (IC) in patients with locally advanced nasopharyngeal carcinoma (NPC).Methods From January 2005 to January 2009,62 cases of locally advanced NPC confirmed by pathological and cytological examination received IC with vinorelbine (25 mg/m2) plus cisplatin (25 mg/m2) for 2-4 cycles and then concurrent chemotherapy and IMRT.Conventional fractionated radiotherapy was adopted in IMRT.The radiotherapy for the nasopharyngeal region was performed a dose of 72-76 Gy/36-38 fractions,and additional 5-Gy gammaknife treatment was carried out in case of local tumor residue.Prophylactic irradiation to the cervical lymph nodes was performed at a dose of 50 Gy,and the dose was increased to 60-70 Gy in case of lymph node enlargement.Results The follow-up rate was 100％.The patients showed a response rate (RR) of 89％ in the nasopharyngeal region and an RR of 90％ in the cervical lymph nodes.The 1-,2-,and 3-year overall survival rates,disease-free survival rates,local relapse-free survival rates,and distant metastasis-free survival rates were 97％,92％,and 82％,94％,73％,and 65％,97％,89％,and 87％,and 97％,84％,and 77％,respectively.The incidence rates of grade 3-4 acute reactions were 37％ for leucopenia,18％ for thrombocytopenia,and 6％ for mucositis.No grade 3-4 long-term temporomandibular joint injury and xerostomia were observed.Conclusions Concurrent chemotherapy and IMRT following IC with vinorelbine (25 mg/m2) plus cisplatin (25 mg/m2) have tolerable adverse effects and can achieve high survival rate in the patients with locally advanced NPC.

OBJECTIVETo optimize the matrix formulation of the effective part Cataplasm of Pogostemon Cablin.

METHODThe optimal preparation prescription was selected by U17 (17(11)) uniform design,and the tacking strength, cohesive strength and transdermal speed constant were used as test indexes. The equations of three test indexes were established by SPSS. With analysis of the contribution of factors by SPSS regression, the optimal matrix formulation was acquired.

RESULTThe optimal matrix formulation is carbopol U10-NoveriteTM7s-glycerine-sorbitol-kaolin-citric acid-aluminum trichloride (1.0:5.0:20:2.0:2.0:0.25:0.2).

CONCLUSIONThe matrix has good adhesive property, proper drug release rate, desirable hemocompatibility with the extractions of Pogostemon cablin.

Adhesiveness ; Biocompatible Materials ; chemistry ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Lamiaceae ; chemistry ; metabolism ; Regression Analysis ; Skin Absorption ; Volatilization

OBJECTIVEThe objective of this article is to summarize the development of evaluation and treatment of posterior malleolus fracture (PMF).

DATA SOURCESData used in this review were mainly from English literature of PubMed data base. Study selection Articles were included in this review if they were related to the PMF or trimalleolar fracture.

RESULTSNo consensus was found regarding what sizes of posterior malleolus fragments would lead to ankle instability thus affecting prognosis and should be fixed. X-ray measurement is unreliable, while CT scan is widely recommended and it can recognize the occult posterior malleolus fractures associated with tibia shaft fractures, which are always undetected previously. Direct posterior malleolus fixation is suitable to stabilize syndesmotic injury. The basic and clinical researches support direct reduction and buttress plate fixation of posterior malleolus fracture through the posterolateral approach. Operative indications and timing of weight bearing are still in discussion.

CONCLUSIONSKnowing whether ankle instability occurs and the proper methods to diagnose, evaluate, and operate can help manage the fracture. Further biomechanical research on ankle stability and clinical study to compare various treatment methods are required.

Ankle Injuries ; surgery ; Fractures, Bone ; surgery ; Humans ; Orthopedic Procedures ; methods ; Tibial Fractures ; surgery

OBJECTIVETo study the effect of laminarin sulphate (LAMS) on the expressions of PTEN and P27kip1 in androgen-independent prostate cancer PC-3 cells in vitro, and investigate the mechanism of its anti-tumor action.

METHODSThe inhibitory effects of different concentrations of LAMS (0, 50, 100, 200 microg/ml) on androgen-independent prostate cancer PC-3 cells were detected by WST-8 assay. The morphology of PC-3 cells was observed under the fluorescence microscope, and the cell cycle and apoptosis were analyzed by flow cytometry. The mRNA and protein levels of PTEN and P27kip1 were measured by RT-PCR and Western blot.

RESULTSLAMS inhibited the proliferation of androgen-independent prostate cancer PC-3 cells in a dose- and time-dependent manner, and the cell cycle analysis showed that PC-3 cells were arrested in the S phase after treated with different concentrations of LAMS. The rate of apoptosis was increased and many typical apoptotic morphological features were observed under the fluorescence microscope. The PTEN and P27kip1 expressions at mRNA and protein levels were increased in a dose-dependent manner.

CONCLUSIONLAMS can inhibit the proliferation, arrest the cell cycle in the S phase and induce apoptosis of prostate cancer PC-3 cells. The significantly increased expressions of PTEN and P27kip1 may be one of the mechanisms for LAMS inhibiting prostate cancer PC-3 cells.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; PTEN Phosphohydrolase ; genetics ; Polysaccharides ; pharmacology ; Prostatic Neoplasms ; blood ; genetics ; RNA, Messenger ; genetics

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Prolymphocytic, B-Cell ; diagnosis ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo study genetic polymorphisms of the KIR2DS4 gene among ethnic Hans from southern China.

METHODSGenomic DNA was isolated from 306 unrelated individuals and amplified with KIR2DS4-specific PCR primers. KIR2DS4-positive samples were genotyped for the entire coding sequence by sequencing-based typing (SBT). Assignment of allelic genotypes was accomplished by using Assign 3.5 software. For samples with inconclusive SBT results, RT-PCR products covering the entire coding sequence of the KIR2DS4 gene were subjected to cloning and haplotype sequencing.

RESULTSAmong all tested samples, 297 were demonstrated to have carried the KIR2DS4 framework gene. For KIR2DS4-positive samples subjected to SBT for the entire coding sequences, no background was observed with the obtained sequences. Three of the seven identified alleles were of novel types, which were officially named by the KIR subcommittee of the World Health Organization Nomenclature Committee for Factors of HLA System. The observed frequencies for the 7 alleles were KIR2DS4*00101 (78.8%), *003 (10.5%), *004 (16.0%), *010 (23.2%), *017 (0.3%), *00105 (0.3%) and *018 (0.7%), respectively. Allele KIR2DS4*007 was not found. The overall frequency for normal cell-surface expression KIR2DS4 alleles including 2DS4*00101, *017 and *00105 was 79.4%, and that for non cell-surface expression alleles including 2DS4*003, *004, *010 and *018 was 50.4%. The ratio between the two was 1.6:1.

CONCLUSIONThe present study has elucidated the allelic diversity of KIR2DS4 among ethnic Hans from southern China, which may provide valuable data for transplantation as well as studies on KIR-associated disease and evolution.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Gene Frequency ; Genotype ; Genotyping Techniques ; methods ; Haplotypes ; Humans ; Polymorphism, Genetic ; Receptors, KIR ; genetics ; Sequence Analysis, DNA ; methods

OBJECTIVEFragile X syndrome (FXS) may be identified by many methods, such as PCR assay and Southern blot. However, each method has its limits or shortcomings. This study explored the reliability of the rapid, convenient and inexpensive hair root fragile X mental retardation protein (FMRP ) assay in the identification of FXS.

METHODSFMRP in hair roots was determined by immunohistochemistry assay in 80 healthy children, in 40 children with mental retardation of unknown etiology and in 12 family members in one pedigree of FXS. FXS was confirmed by 7-deza-dGTP PCR.

RESULTSThere was a high expression of FMRP in hair roots (> or =80%) in healthy children. Two children were confirmed with FXS by 7-deza-dGTP PCR in 40 children with mental retardation of unknown etiology. FMRP expression was 10% and zero respectively in the two children. The other 38 children had FMRP expression of more than 80%. FMRP was not expressed in the two cases of FXS from the pedigree of FXS.

CONCLUSIONSInexpensive, rapid and convenient hair root FMRP assay is reliable for the diagnosis of FXS and may be widely applied for screening and diagnosing FXS in children with mental retardation.

Adolescent ; Child ; Child, Preschool ; Female ; Fragile X Mental Retardation Protein ; analysis ; Fragile X Syndrome ; diagnosis ; genetics ; Hair ; chemistry ; Humans ; Infant ; Male ; Polymerase Chain Reaction

OBJECTIVETo investigate the clinical features, mutation of the GNAS1 and pathogenesis of progressive osseous heteroplasia (POH).

METHODThe typical clinical, pathological and radiographic features of a boy with POH were collected and summarized following family survey. The GNAS1 gene sequence of all family members were amplified by polymerase chain reaction (PCR) and the products were sequenced directly to identify the mutations. A literature review and long-term follow up were also conducted.

RESULTThe patient was an 11-year-old boy who had the onset in infancy, which indicates a chronic progressive cause of disease. The clinical features include the unsmooth local skin of the right shank where spread many rigid rice-like or irregular slabby uplifts, slabby bone-like sclerosis on the left lower mandible, left masticatory muscles, in lateral subcutaneous site of left hip joint and deep tissue, accompanied by gradually progressive difficulty in opening mouth. Histopathology showed that there were loosened hyperplasia of fibroblast and interstitial edema with punctiformed ossification. Radiographs showed flocculence hyperdense image in the subcutaneous tissues and muscles around left lower mandible, and the left masticatory muscles were obviously involved. The 3-dimensional computed tomography showed dislocations of the left temporomandibular joint. Sheeted hyperdense image with inequable density could be noted in lateral muscles of the left hip. And lamellar hyperdense image parallel to the long axis of the bone could be seen in the subcutaneous dorsum of the left foot and achilles tendon. Macro-thumb and of brachydactylia of the hands and feet were not present. The level of calcium, phosphorus and alkaline phosphatase in the blood were normal. Brother of same father but different mothers was free of the disease and no patient of the same disease was found in maternal line and paternal lines. A mutated allele in exon 7 and a polymorphism in exon 5 were found in GNAS1 gene in both of the patient and his father.

CONCLUSIONThere is possibility/likelihood/probability that Chinese children could develop POH. Translocated dermal ossification began in infancy and shows a progressive cause in childhood. The disease is characterized by the heterotopic ossification of the skin, deep tissue, muscles and facial surface tissues. The location of the mutation in this study was different from that reported in abroad studies although exist in the same exons.

Child ; Chromogranins ; DNA Mutational Analysis ; Exons ; GTP-Binding Protein alpha Subunits, Gs ; genetics ; Humans ; Male ; Mutation ; Ossification, Heterotopic ; diagnosis ; genetics ; pathology ; Pedigree