Purpose To investigate the effect of subcellular location of tumor BRCA1 on the sensitivity to ionizing radiation (IR) and PARP inhibitor.Methods siRNA of BRCA1 were first used to inhibit endougenous BRCA1 expression in MCF7 cells.Then,plasmids of pCMV-3xFlag-WT-BRCA1,pCMV-3xFlag-NES-BRCA1 and pCMV-3xFlag-NLS-BRCA1 were transfected in MCF7 cells.Immunofluorescence staining was used to detect BRCA1 subcellular location as well as the formation of Rad51 and γ-H2AX foci.Apoptotic cells were analyzed by flow cytometry,and colony formation assay was performed to evaluate the survival of cells.Results There were 47％ cells with nuclear BRCA1,23％ cells with cytoplasmic BRCA1 and 30％ cell with mixed nuclear and cytoplasmic BRCA1 expression in WT-BRCA1 transfected cell.There were 87％ cells with nuclear BRCA1 in NES-BRCA1 transfected cell,and 82％ cells with cytoplasmic BRCA1 in NLS-BRCA1 transfected cell.There were 87％,84％ and 13％ Rad51 foci positive cells at 2 hours after 4 Gy radiation treatment and 22％,25％ and 59％ γ-H2AX foci positive cells at 24 hours after 4Gy radiation treatment in WT-BRCA1,NES-BRCA1 mutant and NLS-BRCA1 mutant transfected cell respectively.ABT-888 and radiation treatment induced more apoptosis and fewer colonies in NLS-BRCA1 transfected cell than WT-BRCA1,NES-BRCA1 mutant transfected cell.Conclusion Subcellular location of BRCA1 might affect homologous recombination repair of DNA double strand breaks and can be used to predict sensitivity to IR and PARP inhibitor.

Animals ; Cytokines ; blood ; Endotoxemia ; blood ; therapy ; Endotoxins ; blood ; toxicity ; Enzyme-Linked Immunosorbent Assay ; Hemofiltration ; Interleukin-1 ; blood ; Interleukin-10 ; blood ; Models, Animal ; Swine ; Time Factors ; Tumor Necrosis Factor-alpha ; analysis

Objective To investigate the expression of Caspase-9 and heat-shock protein-90 (HSP 90) in rats after focal cerebral ischemia-reperfusion injury.Methods The male SD rats (200-250 g) were divided into three groups by the random number table: normal group, sham group and cerebral ischemia-reperfusion (CIR) group.Each group was sorted into four subgroups including group 6 h, group 24 h, group 48 h and group 72 h according to the reperfusion time.Suture-occluded method was adopted to prepare focal cerebral ischemia-reperfusion(CIR) injury in rat model.Enzymelinked immunosorbent assay (ELISA) method was used to detect the variations of Caspase-9 and HSP-90 expression in rats.Results The changes in Caspase-9 and HSP 90 expression in the brain cells were observed by ELISA method.The expression of Caspase-9 and HSP-90 was weakly expressed in sham group, and was at peak in CIR group within 24 h-48 h, then began to decline at 72 h after the reperfusion time.The differences in the expression of caspase-9 and HSP-70 between sham group and normal group were not statistically significant.Conclusions Apoptotic cells gradually increase along with reperfusion time and reach the peak at 48 h after cerebral ischemia-reperfusion.In ischemia half dark stripe, the expression of Caspase-9 and HSP 90 is increased in neuronal cells after cerebral ischemia-reperfusion, and the positive cells number is at peak at 48 h after cerebral ischemiareperfusion.Apoptosis of neuronal cells after cerebral ischemia and reperfusion is a dynamic evolutionary process.The expression of Caspase-9 and HSP 90 in nerve cells plays an important role in regulating cell apoptosis.

Objective To observe the B-scan ultrasonic characteristics of the posterior vitreous detachment (PVD) and the relationship between the PVD and retina. Design Retrospective case series. Participants 991 cases (1107 eyes) with PVD. Methods The results of B-scan ultrasonography were retrospectively in PVD patients and according to the adhesion between the posterior vitreous cortex (PVC) and eyeball wall, complete PVD and incomplete PVD were divided. Main Outcome Measures The ratio of types and characteristics of PVD. Results 710 eyes(64.1%)were diagnosed as complete PVD, in which 69 eyes(17.3%)accompanied with peripheral ruptured detachment of retina, and 16 eyes(23.0%)PVC adhered with lid of peripheral retina hole. 397 eyes(35.9%)were diagnosed as incomplete PVD, in which 159 eyes(40.0%)as partly PVD, and 76 eyes(47.7%)concomitanted with PVC proliferation and vitreous incompletely splitting. 121 eyes(30.5%)PVC were adhered with the macula, and 117 eye(29.5%)with the optic disc. Conclusions Ultrasonography can be applied to diagnose PVD precisely,which can provide credible morphologic evidence for the clinic.

OBJECTIVETo observe the changes of bone histomorphometry parameters in condyle of rabbits during mandibular forward positioning.

METHODSForty rabbits with eight weeks of age were simple randomly divided into the experimental group (n = 24) and control group (n = 16).Mandibles of rabbits in the experimental groups were induced to forward position by a functional appliance.The experimental group and control group were sacrificed after 2 and 4, 8, 12 weeks.The specimens from right tempromandibular joints were processed for undemineralised sections.These sections were used for fluorescent microscopy observation and the measurement of bone histomorphometry.

RESULTSAt 8 week bone-volume fraction [(75.83 ± 1.10)%], trabecular thickness [(103.28 ± 2.89) µm], trabecular number [(2.86 ± 0.06) mm(-1)], mineral apposition rate [(2.32 ± 0.02) µm/d] and index of osteoblast [(30.20 ± 0.47) N/mm(2)] in subchondral region of the cartilage in the experimental group were significantly increased compared with the age-matched controls [respectively (64.00 ± 1.54)%, (87.00 ± 1.13) µm, (1.84 ± 0.08) mm(-1), (1.69 ± 0.02) µm/d and (21.07 ± 0.59) N/mm(2)] (P < 0.05). However, trabecular separation [(170.00 ± 2.25) µm] was lower than those in the controls [(241.50 ± 1.57) µm] with significant difference(P < 0.05). There was no significant difference in the bone histomorphometry parameters of the central region between the experimental group and the age-matched control group (P > 0.05).

CONCLUSIONSThe pattern of bone histomorphometry parameters expression in subchondral region has a high correlation with the adaptive remodeling of the condyle after functional appliance.

Animals ; Cartilage ; anatomy & histology ; growth & development ; Female ; Male ; Mandibular Advancement ; Mandibular Condyle ; anatomy & histology ; growth & development ; Orthodontic Appliances, Functional ; Rabbits ; Random Allocation

Objective To analyze the cost of capsule endoscopy in diagnosing small bowel bleeding and to compare it with traditional diagnostic methods.Methods The patients suspected with small bowel bleeding were divided into group A(n=58,collected during 1998 to 2005)diagnosed with traditional processes and group B (n=93,collected during 2002 to January 2005)diagnosed with capsule endoscopy.The diagnostic yield,specific treatments,examination costs and other accumulated costs of two groups was compared.The examination cost ratio and the integration cost ratio were evaluated.The sensitivity analysis was performed.Results The diagnostic yield of small bowel bleeding in group A and group B were 22.4%(13/58) and 86%(80/93),respectively.The total of examination costs were 133 750 RMB and 790 500 RMB,respectively.The examination costs in group B(RMB 9881.3/each) was slightly lower than group A(RMB 10 288.5/each).Furthermore,as the diagnostic yield of group B was significantly higher than group A(P=0.001).The specific treatments based on the results of the diagnosis was 37.4% higher in group B(49.5%) than group A(12.1%).That means the cost of repeat- ed consultations,emergencies room visit,examinations,supporting treatments and hospitalizations in group B were significantly decreased.After the adjustment,the cost in group B(RMB 9881/patient) was lower than group A(16 361.5 RMB in one month—97 424.0 over 5 years/patient).The total cost of each patient in group A was 6480.2—87 542.7 RMB more than group B,which represented 1.7—9.9 folds increase.Conclusions The patients who suspected with small bowel bleeding and had a negative results of gastroscopy and colonoscopy were recommended to have capsule endoscopy which yields early diagnosis and less cost.

OBJECTIVETo determine the incidence of work-related acute pesticide poisoning among farmers in Jiangsu Province and to identify the risk factors.

METHODSMulti-stage stratified sampling and cluster sampling methods were used to randomly select 1490 farmers from the south (Z village), centre (Y village) and north (X village) of Jiangsu Province. The database was established by EpiData 3.1. SPSS 17.0 and SAS 9.13 were used to do chi-square test, trend chi-square test, single factor and multi-factor logistic regression analysis.

RESULTSAmong 1490 farmers, 121 people had work-related acute pesticide poisoning in the past year. The incidence rate in Jiangsu Province was 8.1%. 118 farmers (97.5%) were mild pesticide poisoning.3 farmers (2.5%) were moderate pesticide poisoning. The incidence rate of Work-related acute pesticide poisoning in X village of North Jiangsu Province (13.5%, 68/505) is obviously higher than that in Y village of Central Jiangsu Province (4.8%, 19/399) and Z village of South Jiangsu Province (5.8%, 34/586), and the difference is statistically significant (χ(2) = 29.60, P < 0.01). The incidence rate of Work-related acute pesticide poisoning in female (10.4%, 58/557) is obviously higher than that in male (6.8%, 63/933), and the difference is statistically significant (χ(2) = 6.26, P < 0.05). There are lack of safety guidance, unsafety application methods (without alternate row spraying, without backward application, without down-wind application) and risk behaviors in the spraying (without read labels, preparing pesticides without gloves, wiping sweat with hand (s), equipment leakage, body pollution by pesticide, working when feeling sick, without bath after work). If farmers have risk behaviors above, the incidence rate of Work-related acute pesticide poisoning will be obviously higher than farmers who receive safety guidance and have no such dangerous behaviors, and all the differences are statistically significant (P < 0.05). The group of 24 ∼ 34 years old has the highest risk of work-related acute pesticide poisoning (OR = 7.15).

CONCLUSIONThe incidence rate of work-related acute pesticide poisoning among farmers in Jiangsu Province was higher. Effective measures are needed to prevent and reduce the occurrence of work-related acute pesticide poisoning. We can teach the farmers who spray pesticides about safety guidance, safety application methods and avoiding risky behaviors.

Adult ; Aged ; Agricultural Workers' Diseases ; epidemiology ; Agriculture ; methods ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Occupational Exposure ; Pesticides ; poisoning ; Risk Factors

Bao-Xie-Ning (BXN), a traditional Chinese herbal medicine (CHM) formula composed of Fructus Evodiae, Flos Caryophylli and Cortex Cinnamomi, and used for the treatment of infant diarrheal illness, was subject to systematic assessment for its putative multiple pharmacodynamic effects and pharmacological antidiarrheal mechanisms.

OBJECTIVETo investigate the genetic polymorphism and applied value in forensic medicine of five short tandem repeats (STRs) loci(DXS7130, DXS7132, DXS7133, DXS6804 and DXS8378) on X-chromosome in Achang ethnic group.

METHODSAllele frequency and genotype distribution of five X-STR loci among 100 unrelated individuals from Achang ethnic group were analyzed by PCR following polyacrylamide gel electrophoresis and silver staining. Hardy-Weinberg equilibrium of females was tested and forensic parameters were calculated.

RESULTAll the five STR loci were polymorphic in Achang ethnic group. Chi-square test indicated that genotype distribution of females was in accordance with Hardy-Weinberg equilibrium.

CONCLUSIONThese results enrich Chinese genetic database and can be applied to individual identification, paternity testing and population genetics.

Alleles ; China ; Chromosomes, Human, X ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic

OBJECTIVETo prepare a glypican-3 (GPC3)-targeting hepatocellular carcinoma MR molecular probe and to evaluate its targeting specificity using HepG2 cells.

METHODSPoly(lactic-co-glycolic acid) (PLGA) nanoparticles were prepared by a double emulsion solvent evaporation method, and the surfaces were connected with anti-GPC3 mono-antibody and paramagnetic substance Gd3+. The physical properties of the probes were investigated using fluorescence microscopy, electron microscopy, Malvern particle size analysis, inductively coupled plasma atomic emission spectroscopy (ICP-AES) and 1.5T MR imaging. The specificity of the probes to target cultured HepG2 cells was determined by laser confocal microscopy. The signal characteristics, including signal-to-noise ratio (SNR), after co-incubation with HepG2 cells were analyzed by 1.5T MR imaging. Significance of differences between multiple groups (target group, non-target group, and control group) was assessed by one-way analysis of variance, and between two groups was assessed by the LSD-t test. A difference was considered to be statistically significant at P less than 0.05.

RESULTSThe GPC3-targeting hepatocellular carcinoma MR molecular probes were successfully prepared. The nanoparticles had a spherical shape, size of 495 +/- 17.5 nm, uniform size distribution, good dispersibility, no obvious aggregation, and could significantly increase the T1 signal. Using the ICP-AES measurement, 1 mol PLGA carried about 12 mol Gd3+, and as the Gd3+ concentration increased, the T1 signal increased. The prepared MR molecular probes could specifically target HepG2 cells, and could increase the T1 signal. The SNR value of the target group was 3.45 +/- 0.21, of the non-target group was 1.43 +/- 0.07, and of the control group was 1.12 +/- 0.03. The SNR value of the target group was significantly higher than that of the non-target group and the control group (P less than 0.05); there was no significant difference between the non-target group and the control group (P more than 0.05).

CONCLUSIONPLGA nanoparticles, anti-GPC3 mono-antibody and paramagnetic Gd3+ can be used to successfully prepare GPC3-targeting hepatocellular carcinoma MR molecular probes which are capable of specifically targeting HepG2 cells in vitro and being detected by 1.5T MR imaging. These MR molecular probes may represent a useful noninvasive imaging method for detecting early hepatocellular carcinoma in vivo.