Humans ; Occupational Diseases ; epidemiology ; prevention & control ; Risk Assessment

Fanconi anaemia (FA) is an autosomal recessive inherited disorder caused by defects in hematopoietic stem cells. The clinical manifestations of FA are diverse and complicated. FA cells display high hypersensitivity to agents which produce interstrand DNA cross-links such as mitomycin C (MMC) or diepoxybutane (DEB). At least eight complementation groups with defects in eight genes (FANCA, FANCB, FANCC, FANCD(1), FANCD(2), FANCE, FANCF and FANCG) have been identified by gene analysis. Six genes (corresponding to subtypes A, C, D(2), E, F and G) have been coloned, and the encoded FA proteins interact in a common cellular pathway - "FA Pathway", through which modulate DNA repair. The progress of research on FA molecular mechanism provides gene therapy of FA with theory basis. FA cells transduced with the use of retrovirus carring the normal FA gene cDNA manifestate phenotypic correction of hypersensitivity to DNA cross-linking agents, such as MMC. In this review the clinical manifestations and gene composition of FA, and the functions of encoded FA proteins were summarized. The hematopoietic stem cell transplantation and gene therapy for FA patients were discussed.
Cell Cycle Proteins ; DNA-Binding Proteins ; Fanconi Anemia ; genetics ; metabolism ; therapy ; Fanconi Anemia Complementation Group C Protein ; Fanconi Anemia Complementation Group D2 Protein ; Fanconi Anemia Complementation Group Proteins ; Genetic Therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; Mutation ; Nuclear Proteins ; genetics ; Proteins ; analysis ; genetics

The study was purposed to investigate the effects and mechanism of bone marrow-derived mesenchymal stem cells (MSCs) on graft-versus-host desease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The model of GVHD in rat had been established by allo-HSCT with donor derived T cells. The occurence of GVHD in recipients was observed in condition with or without donor derived MSC co-transplantation. Effects of MSCs on GVHD were analyzed by model rat survival rate and pathology. Proportions of CD4+CD25+ regulatory T cells were determined by using label spleen lymphocytes and thymocytes with double fluorescent-labeled antibodies and flow cytometry. The results showed that MSCs inhibited the lethal GVHD after HSC co-transplantation and increased the survival rate. The ratio of CD4/CD8 deceased in GVHD group in different levels, as compared with that in the experimental group. The proportion of CD4+CD25+ regulatory T cells of spleen lymphocytes was 31.55 +/- 7.58% and 20.90 +/- 1.90% in experimental and GVHD groups, respectively. Similarly, the proportion of CD4+CD25+ regulatory T cells of thymocytes was 93.20 +/- 2.69% and 57.17 +/- 6.79% in experimental and the GVHD groups, respectively. Meanwhile the proportion of CD4+CD25+ regulatory T cells was higher in experimental group than that in GVHD group. It is concluded that MSCs may prevent the lethal GVHD after allo-HSC co-transplantation and raise the survival rate of model rats by acting on the CD4+CD25+ regulatory T cells in vivo.
Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; adverse effects ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Graft vs Host Disease ; immunology ; Interleukin-2 Receptor alpha Subunit ; immunology ; Mesenchymal Stromal Cells ; immunology ; physiology ; Rats ; Rats, Inbred F344 ; Rats, Wistar ; T-Lymphocytes, Regulatory ; immunology

The hydroxycamptothecin (HCPT) PEGylated liposomes (HCPT-LP) were modified with RGD cyclopeptide formed the tumor-targeting liposomes (HCPT-RGD-LP). HCPT-LP and HCPT-RGD-LP were injected intravenously with single dose of 5 mg x kg(-1) to rats. The drug concentration in plasma was determined and the pharmacokinetic behaviour was compared. The HCPT distribution in heart, liver, spleen, lung, kidney and plasma of mice was investigated following intravenous administration of HCPT-LP and HCPT injection. The nude mice implanted human hepatoma HepG2 cells were studied by in vivo imaging. The fluorescent probe was DiR and the nude mice were injected with DiR PEGylated liposomes (DiR-LP) and DiR-LP modified with RGD cyclopeptide (DiR-RGD-LP). The results showed that there was no significant difference (P > 0.05) of main pharmacokinetic parameters t1/2beta, CL, V(c), AUC(0-48 h), AUC(0-inifinity), MRT(0-48 h), MRT(0-infinity) between HCPT-RGD-LP and HCPT-LP. HCPT-LP had a remarkably better long-circulating effect than HCPT injection in mice and the concentration of HCPT was highest in liver. The DiR accumulation in tumors of DiR-RGD-LP was higher than that of DiR-LP by the visualized fluorescence of in vivo imaging. It indicated that such PEGylated liposomes modified with RGD cyclopeptide could improve the tumor targeting efficacy.
Animals ; Area Under Curve ; Camptothecin ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacokinetics ; Diagnostic Imaging ; Drug Delivery Systems ; Female ; Fluorescent Dyes ; Hep G2 Cells ; Humans ; Liposomes ; administration & dosage ; chemistry ; pharmacokinetics ; Liver Neoplasms ; diagnosis ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oligopeptides ; administration & dosage ; chemistry ; pharmacokinetics ; Polyethylene Glycols ; administration & dosage ; chemistry ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectroscopy, Near-Infrared ; Tissue Distribution

BACKGROUNDDense congenital cataracts often cause severe visual impairment. The results of long-term follow-up of dense bilateral congenital cataract in China have not been well documented. The purpose of this study was to evaluate the long-term visual function in children who underwent cataract extraction for dense bilateral congenital cataract in southern part of China.

METHODSMedical records of children who underwent surgery of dense bilateral congenital cataract between January 1992 and December 2000 at Zhongshan Ophthalmic Center of Sun Yat-sen University were retroactively reviewed. In 38 children available for current follow-up, best corrected visual acuity (BCVA) and stereoscopic vision, as well as nystagmus, strabismus, and other complications, were evaluated. The mean follow-up period was 107.6 months (range 60 to 167 months).

RESULTSThe mean age of cataract extraction and secondary intraocular lens implantation were 5.6 months (range 3 to 12 months) and 4.2 years (range 2.4 to 15 years), respectively. The mean BCVA was 0.25 in the better eye and 0.16 in the fellow eye. Stereoscopic vision was absent in all patients, and 3 children had simultaneous perception. Nystagmus was detected in all cases and strabismus in 35 cases. A high correlation was found between timing of cataract extraction and final BCVA of the better eye (r = -0.55, P = 0.00). A statistically significant difference was found in BCVA between post- and pre-treatment of amblyopia (t = 5.65, P = 0.00).

CONCLUSIONSLong-term visual function in children with dense bilateral congenital cataract was poor when cataract surgery was performed at age of 3 months or later. Earlier cataract surgery with adequate optical rehabilitation contributed to better visual outcome.

Adolescent ; Cataract ; congenital ; Cataract Extraction ; Child ; Child, Preschool ; Follow-Up Studies ; Humans ; Infant ; Retrospective Studies ; Visual Acuity

OBJECTIVETo explore the effect of Bcl-xl on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis signal pathway and apoptosis.

METHODSA dominant negative mutant of ikB (pmi kappaB) and Green Fluorescent Protein (GFP) expression plasmid pEGFP-C1, pmi kappab and pEGFP-C1 and Bcl-xl expression construct pBcl-xl/HA, were co-transfected into HeLa cells. Expression plasmid pBcl-xl/HA was introduced into p65-/-MEF cells in which nuclear factor-kappaB (NF-kappaB)/p65 was deficient, to establish cell line p65-/-Bcl-xl expressing Bcl-xl by selection with puromycin. These cells were treated with TNFalpha at a concentration of 10 ng/ml, and apoptotic cell death was examined microscopically with trypan blue staining. The proteins were abstracted from treated cells, and caspase 8 activation and cleavage of poly (ADP-ribose) polymerase (PARP) were examined by western blot using a specific antibody that recognized cleaved caspase 8 and cleaved PARP, respectively.

RESULTSHeLa cells transfected with pmi kappaB, TNFalpha showed significant cell death as they became rounded, shrank, and detached. However in HeLa cells co-transfected with pBcl-xl and pmi kappaB, no cell death was observed after treatment with TNFalpha. In p65-/- MEF cells; cell death was observed at 4 hours after treatment with TNFalpha, and cell death reached 90% at 12 hours after the treatment. However, in p65-/-Bcl-xl/HA cells expressing Bcl-xl, no cell death was seen even when treated with TNFa for 24 hours. Meanwhile, in pmikB/HeLa cells transfected with pmi kappaB, TNFalpha induced caspase 8 activation and PARP cleavage, but in the HeLa cells co-transfected with pBcl-xl and pmi kappaB, no activated caspase 8 and cleaved PARP were observed after treatment with TNFalpha.

CONCLUSIONIn the experimental system in which NF-kB was inhibited, Bcl-xl blocked TNFalpha-induced apoptosis signal pathway and apoptosis. These results bring to light that further studies of the pathogenesis and therapy of TNFa-related diseases are needed.

Apoptosis ; drug effects ; Caspase 8 ; metabolism ; HeLa Cells ; Humans ; NF-kappa B ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; bcl-X Protein ; pharmacology

OBJECTIVETo investigate the screening methods for identifying the populations susceptible and resistant to noise-induced hearing loss (NIHL) and to provide a reference for future research.

METHODSWorkers who were exposed to 75 ∼ 120 dB noise in enterprises were included in the study. Field investigation of occupational health was conducted; workers' basic information and data on hearing threshold levels were collected. Paired chi-square test was used to compare each two of three screening methods, which were used at home and abroad to identify noise-susceptible and noise-sensitive populations, in terms of noise exposure level, general information, and noise-induced hearing threshold shift.

RESULTSThere were no significant differences in the noise exposure level, basic information, and left and right ears' hearing threshold levels of noise-susceptible and noise-sensitive populations between each two of the three screening methods (P > 0.05), according to the paired chi-square test. However, high-frequency hearing threshold had statistically significant difference among the three methods. As a whole, methods B and C were superior to method A.

CONCLUSIONThe workers in China are younger than before, with more awareness of self-protection, and individual protection is enhanced in them. Currently, method B is more suitable for screening out the population susceptible to NIHL in China.

Adult ; China ; Disease Susceptibility ; Female ; Hearing Loss, Noise-Induced ; diagnosis ; Humans ; Male ; Mass Screening ; Noise, Occupational ; adverse effects ; Surveys and Questionnaires ; Young Adult

Objective To explore the molecular mechanism of Gasdermin B(GSDMB)regulating the fate of intestinal epithelial cells.Methods The human GSDMB plasmid was overexpressed into two human intestinal epithelial cell lines(NCM460 and HT-29 cells)and human colon-derived organoids.Western blotting was used to confirm the efficiency of electroporation.Cell counting kit(CCK8),cell apoptosis,and cell cycle by flow cytometry were performed to analyze the effect of GSDMB overexpression on cell function.Transcriptome sequencing was used to analyze the downstream effector molecules of GSDMB.T test was used to compare the data between the two groups.Results The overexpression of GSDMB protein in the two intestinal epithelial cell lines was successfully reconstructed.The absorbance value(A)of human intestinal epithelial cells overexpressing GSDMB protein[NCM460 cells:(1.17±0.01),HT-29 cells:(0.96±0.06)]was significantly lower than that of blank control cells[NCM460 cells:(1.67±0.12),HT-29 cells:(1.24±0.07)](t=7.24 and 5.46,P＜0.05).The number of apoptotic cells in the GSDMB overexpression group[NCM460 cells:(12.03±1.55),HT-29 cells:(29.30±4.48)]was significantly higher than that in the blank group[NCM460 cells:(4.96±1.74),HT-29 cells:(6.95±3.42)](t=5.26 and 6.97,P＜0.05).Cell cycle analysis showed that the ratio of cells at G0/G1 phase in the GSDMB overexpression group[NCM460 cells:(47.98±5.28)％,HT-29 cells:(38.04±3.45)％]was significantly lower than that in the control group[NCM460 cells:(59.54±3.90)％,HT-29 cells:(63.81±1.76)％](t=3.05 and 11.53,P＜0.05).Transcriptome sequencing results showed that the dual specificity phosphatase 4 and 6(DUSP4 and DUSP6)genes were significantly upregulated after GSDMB protein expression.Fluorescence quantitative PCR results confirmed that the relative expression levels of DUSP4(2.45±0.15)and DUSP6(4.34±0.22)in intestinal epithelial cells transfected with GSDMB were significantly higher than those in the control group(1.06±0.05 and 1.01±0.02)(t=15.08 and 26.52,P＜0.05).After GSDMB-expressing NCM460 cells were treated with the DUSP inhibitor BCI,the BCI treatment group had a significantly increased expression level of p-ERK compared to the control group[(1.14±0.17)vs.(0.58±0.12)](t=5.42,P=0.002);the A value(1.84±0.07)and G0/G1 phase ratio(59.83±2.17)％in the BCI treatment group were significantly higher than those in the non-treatment group[(1.52±0.10)and(52.10±2.23)％],and the number of apoptosis in the BCI treated group(7.60±0.56)was significantly lower than that in the untreated group(12.57±1.00)(t=4.71,4.31,7.52,P＜0.05).TUNEL staining in human colon organoids showed a significant increase in apoptotic cells,and the relative expression level of DUSP6 protein(0.85±0.09)was significantly higher than that of the control group(0.21±0.04),accompanied by a decrease in p-ERK levels[(0.83±0.18)vs.(0.19±0.06)],with statistical significance(t=11.95,P＜0.001;t=6.56,P＜0.001).Conclusion GSDMB may inhibit cell proliferation,induce cell cycle arrest,and promote apoptosis by upregulating dual specificity phosphatase DUSP6-mediated ERK phosphorylation,thus affecting the fate of intestinal epithelial cells.