Adult ; Air Pollutants, Occupational ; adverse effects ; Chemical Industry ; Female ; Humans ; Insecticides ; adverse effects ; Male ; Maximum Allowable Concentration ; Middle Aged ; Occupational Diseases ; chemically induced ; diagnosis ; Organothiophosphorus Compounds ; adverse effects ; Phosphorus Compounds ; adverse effects ; Sulfides ; adverse effects

Objective To evaluate the diagnostic efficiency of double-balloon enteroscopy in small bowel Crohn’s disease. Methods In sixty five patients with suspected small bowel Crohn's disease double-balloon enteroscopy were performed, and some of them received enteroscopy and enteroclysis, capsule endoscopy as well.Results The first enteroscopy was performed via mouth in 20 of 65 cases, and the lesions were detected in 11 cases (55%), 5 of 9 cases(55.6%) had lesions detected in enteroscopy via anus while nothing was found in mouth route. Among 45 cases examed by enteroscopy firstly via anus, 34 cases had lesions detected (75.6%), 8 of 11 cases(72.7%) had lesions found in following exam via mouth. Totally 58of 65 had lesions detected through enterosocpy examination, the overall diagnostic yield was 89.2%. Twenty four of 46 cases had positive findings with enteroclysis. The diagnosis of Crohn's disease was comfirmed in 14 of 22 patients(63.6%) underwent capsule endoscoy. The diagnosis was finally confirmed by enteroscopy only in 11 patients(78.6%).Conclusion The entire small intestine could be examined by enteroscopy with combination of mouth and anus route. Double-balloon enteroscopy was an ideal diagnostic modality for small bowel Crohn's diseases, which was also valuable in assessment on extent and severity of the disease. Small bowel enteroclysis was a useful screening alternative for selecting procedure route in DBE.

Objective To investigate the diagnostic yield and accuracy of double balloon enteroscopy, barium enteroclysis and capsule endoscopy in patients with suspicion of small bowel tumors. Methods Double balloon enteroscopy were performed in fifty nine patients with suspicion of small bowel tumors.The route of enteroscopy could be either via mouth or via anus.At the same time,34 and 17 out of 59 subjects received either barium enteroclysis or capsule endoscopy.The results of exams were analyzed independently and final diagnosis of each case was compared thereafter.Results Nineteen of 34 patients undergone the enteroclysis were diagnosed as small bowel tumor.The diagnostic yield was 55.9%.The diagnosis was finally confirmed by the enteroscopy in 12 cases,which indicated the accurate rate of enteroclysis was 63.2%(12/19).Double balloon enteroscopy detected tumors in 3 of 15 subjects with negative enteroclysis finding.The diagnostic yield of capsule endosocpy was 47.1%(8/17),and among the 8 cases diagnosis was comfirmed by the enteroscopy in 4 cases.Small bowel tumors were detected in 2 of 9 cases with negative capsule endoscopy findings.Thirty-six cases of small bowel tumor were detected by double balloon enteroscopy via a route(mouth or anus),and 16 patients were diagnosed after both route procedure.No small bowel tumor was found in 7 paitents.The overall diagnostic yield of enteroscopy was 88.1%.The diagnosis were all finally confirmed by pathological examination.No procedure-related complication were observed.Conclusion Double balloon enteroscopy is superior to enteroclysis and capsule endoscopy in diagnostic yield and accuracy for small bowel tumors.

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

Objective To develop an intelligent flight examination information management system to realize informatized flying personnel physical examination.Methods The system involved in Java Web technology for system design and etmvc framework and MySQL database for background development.A physical examination instrument acquired and received data with Android system,and then transmitted them to the information management system.The information management had open interfaces for data transmission and interaction.Results The system could complete physical examination of hundreds of flying staffs in short time,and then upload the physical examination data to the flight surgeon department for data interaction,storage and statistical analysis.Conclusion The system gains advantages in convenient flying personnel physical examination and rapid information transmission and utilization,and thus is worthy promoting practically for flying personnel physical examination.

OBJECTIVETo explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats.

METHODSSixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated.

RESULTSDutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05).

CONCLUSIONDutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and β-catenin expressions, and damaging epididymal epithelial cell junctions.

Animals ; Azasteroids ; pharmacology ; Claudin-1 ; metabolism ; Dihydrotestosterone ; blood ; Dutasteride ; Epididymis ; drug effects ; metabolism ; Female ; Fertility ; drug effects ; Humans ; Intercellular Junctions ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Testosterone ; blood ; Urological Agents ; pharmacology ; beta Catenin ; metabolism

Objective To investigate the effects and mechanisms of receptor-interacting serine-threonine kinase 3 (RIP3) in the formation of cholestatic hepatic injury.Methods The experimental study was conducted.(1) Processing and viability of hepatic stellate cell line HSC-T6:HSC-T6 cells were transfected by RIP3-siRNA and NC-siRNA,respectively.The viabilities of un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively measured by cell-counting kit-8 (CCK-8).HSC-T6 cells were treated by 100 μmol/L Glycochenodeoxycholic acid (GCDCA) at 0,2,4,8 and 12 hours,and then were extracted and stored,12-hour cell viability was measured by CCK-8.RIP3 that was treated by 100 μmol/L GCDCA knocked down HSC-T6 cells to establishment RIP3 knockdown HSC-T6 cells (RIP3-KD cells).RIP3-KD cells were cultured for 12 hours,and cell viability was measured.(2) Mice model of bile duct ligation (BDL):40 adult mice were randomly divided into 8 groups,5 mice in each group.Sham group:bravery manager was only separated,without ligation,and bloods of inferior vena cava and liver tissues were extracted at 7 days postoperatively.The BDL-1,-3,-5,-7,-14,-21 and-28 d groups:bloods of inferior vena cava and liver tissues were extracted at 1,3,5,7,14,21 and 28 days postoperatively,respectively.(3) The relative expressions of RIP3,α-SMA and TNF-αmRNA in the cells and liver tissues were detected by quantitative real-time polymerase chain reaction (RT-PCR).(4) The relative expressions of RIP3,α-SMA and TNF-α proteins were detected by Western blot.Measurement data with normal distribution were represented as-x±s.The ANOVA was used for data analysis in different time gradient.Comparisons among groups were analyzed using the ANOVA.Pairwise comparison was done by the t test.Results (1) The HSC-T6 cells viability and expressions of RIP3,α-SMA,TNF-α mRNA and proteins:results of CCK8 test showed that 12-hour viabilities of GCDCA-treated HSC-T6 cells,GCDCA-treated RIP3-KD cells,HSC-T6 cells and RIP3-KD cells were 61.3％ ±0.3％ and 83.2％ ±0.4％ and 98.4％ ±0.7％ and 97.4％ ±0.7％ respectively,showing statistically significant differences in the viabilities among them (F =115.200,P＜ 0.05),and showing no statistically significant difference in the viabilities between HSC-T6 cells and RIP3-KD cells (t =1.283,P＞ 0.05).There were statistically significant differences in the viabilities between HSC-T6 cells and GCDCA-treated HSC-T6 cells or GCDCA-treated RIP3-KD cells (t =17.910,6.604,P＜ 0.05) and between GCDCA-treated HSC-T6 cells and GCDCA-treated RIP3-KD cells (t=7.186,P＜0.05).Results of RT-PCR test showed relative expressions of RIP3 mRNA in un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively 0.012 1±0.001 3,0.011 2±0.003 1 and 0.002 8±0.000 5,with a statistically significant difference (F =20.410,P ＜ 0.05).There was no statistically significant difference in relative expressions of RIP3 mRNA between un-transfected and NC-siRNA-transfected HSC-T6 cells (t =0.483,P ＞0.05).The relative expression of RIP3 mRNA in RIP3-siRNA-transfected HSC-T6 cells was significant different from that in un-transfected and NC-siRNA-transfected HSC-T6 cells (t =11.760,4.586,P＜0.05).The relative expressions of RIP3 mRNA,α-SMA mRNA and TNF-α mRNA in GCDCA-treated HSC-T6 cells at 0,2,4,8 and 12 hours were 0.012 1±0.001 3,0.011 2±0.003 1,0.021 2±0.002 2,0.027 8±0.002 1,0.029 8±0.002 3 and 0.571±0.012,0.611±0.024,0.691±0.021,0.711±0.021,0.752±0.031 and 0.873±0.022,0.912± 0.024,1.015±0.031,1.210±0.042,1.471±0.041,respectively,showing an increased trend over time and statistically significant differences (F=70.720,30.050,166.700,P＜0.05).The relative expressions of RIP3 mRNA in HSC-T6 cells and GCDCA-treated HSC-T6 cells were 0.012 1±0.001 3 and 0.029 8±0.002 3,with a statistically significant difference (t=13.970,P＜0.05).Results of Western blot showed that relative expressions of RIP3 protein in un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively 0.054 ± 0.012,0.013 ± 0.008 and 0.052± 0.021,with a statistically significant difference (F =7.410,P＜0.05).There was no statistically significant difference in relative expressions of RIP3 protein between un-transfected and NC-siRNA-transfected HSC-T6 cells (t =0.143,P ＞ 0.05),and statistically significant differences were found in relative expressions of RIP3 protein between RIP3-siRNA-transfected HSC-T6 cells and un-transfected or NC-siRNA-transfected HSC-T6 cells (t =4.924,3.006,P＜0.05).The relative expressions of RIP3,α-SMA and TNF-oα proteins in GCDCA-treated HSC-T6 cells at 0,2,4,8 and 12 hours were 0.045±0.024,0.047±0.034,0.062±0.025,0.121±0.015,0.154±0.034 and 0.064±0.031,0.072±0.017,0.097±0.035,0.078±0.031,0.254±0.051 and 0.078±0.025,0.094±0.037,0.129±0.041,0.198±0.011,0.324±0.061,respectively,showing an increased trend over time and statistically significant differences (F =9.658,15.810,20.090,P＜0.05).The relative expressions of RIP3 protein in HSC-T6 cells and GCDCA-treated HSC-T6 cells at 12 hours were 0.045±0.024 and 0.154±0.034,with a statistically significant difference (t =4.536,P＜0.05).(2) Expressions of RIP3,α-SMA and TNF-α mRNA in hepatic tissues of mice in each group:the results of RT-PCR showed that relative expressions of RIP3 mRNA,α-SMA mRNA and TNF-α mRNA in the Sham,BDL-1 d,BDL-3 d,BDL-5 d,BDL-7 d,BDL-14 d,BDL-21 d,BDL-28 d groups were 0.047 3±0.003 1,0.041 2±0.007 8-0.339 7±0.017 1 and 2.948±0.612,2.654± 1.032-8.387±0.910 and 0.563±0.078,0.610±0.113-1.078± 0.289,respectively,with statistically significant differences (F =25.180,27.820,7.425,P＜0.05).The results of western blot showed that relative expressions of RIP3,α-SMA and TNF-α proteins in Sham,BDL-1 d,BDL-3 d,BDL-5 d,BDL-7 d,BDL-14 d,BDL-21 d,BDL-28 d groups were 0.245±0.011,0.228±0.023-1.018±0.052 and 0.424±0.057,0.392±0.041-0.985±0.081 and 0.551 ±0.052,0.588±0.087-0.962±0.074,respectively,with statistically significant differences (F=19.160,94.410,22.750,P＜0.05).Conclusion Cholestasis promotes hepatic injury and fibrosis by inducing TNF-α pathway activation and upregulation RIP3.

OBJECTIVETo study the variation of specific antibody among convalescent of severe acute respiratory syndrome (SARS) patients through a three-year program.

METHODSSera samples were collected from SARS cases in the 5th, 20th and 35th month after onset of the illness. The SARS-CoV specific antibody was detected for all of them by ELISA and neutralized test simultaneously. The titer of neutralizing antibodies was calculated using Reed-Muench method, and the comparison between different time groups was analyzed regarding the variance of data on repeated measures after logarithm conversion.

RESULTS13, 17 and 13 sera samples were collected in the 5th, 20th and 35th month after onset. Results showed that despite the fact that the positive rates of ELISA antibody were 100%, 82.4% and 84.6% respectively,the neutralizing antibody was still positive for all the samples. The average neutralizing antibody titers were 1:43 (1:16-1:203), 1:36 (1:17-1:59) and 1:21 (1:10-1:39) on the 5th, 20th and 35th month after onset, and the differences were statistically significant (F = 60.419, P < 0.001). On the 35th month after the onset, 30.8% (4/13) of the patients were still having the neutralizing antibody level of above 1:36, but the neutralizing antibody level in another 30.8% (4/13) of the patients had decreased to as low as 1:10, when the cut-off level was set as 1:8.

CONCLUSIONResults of the study indicated that the neutralizing antibody of SARS cases could last for at least three years, but the sera specific antibody in SARS cases decreased gradually when time went by. However, neutralizing antibody in some of the cases decreased to a lower level on the 35th month. Further follow-up study was worthwhile to observe the long-lasting profile of antibody existence on SARS cases.

Antibodies, Neutralizing ; analysis ; Enzyme-Linked Immunosorbent Assay ; Follow-Up Studies ; Humans ; Severe Acute Respiratory Syndrome ; immunology

Adolescent ; Common Variable Immunodeficiency ; complications ; Female ; Humans ; Lung Diseases, Interstitial ; etiology ; pathology ; physiopathology ; Pneumonia ; etiology ; pathology ; physiopathology

BACKGROUNDThe prospects of using immature CD8a(+) dendritic cells (DC2) to establish transplant immunologic tolerance and treatments for autoimmune diseases in the future are promising. However, the methods for inducing DC2 are still being explored. The present study was aimed to investigate the optimal in vitro conditions for preparing large numbers of predominant DC2 from murine bone marrow cells.

METHODSThree groups of bone marrow cells cultured under different conditions were examined, namely a cytokine-induced experimental group (cytokine group), a control group with a low concentration of granulocyte-macrophage colony stimulating factor (GM-CSF, low GM-CSF group) and a control group without endogenous cytokines. The cytokine group was cultured with 5 ng/ml GM-CSF, 25 ng/ml Flt3 ligand (Flt3L), 20 ng/ml interleukin 4 (IL-4) and 100 ng/ml stem cell factor (SCF). The low GM-CSF control group was cultured with 0.4 ng/ml GM-CSF, 25 ng/ml Flt3L and 100 ng/ml SCF, without IL-4. The control group without exogenous cytokines was cultured without additional cytokines. All cells were cultured at 37 degrees C under 5% CO2. On days 3, 7 and 16, 4-color flow cytometry was carried out to analyze the cell phenotypes, and the total cell numbers were counted to analyze the cell yields. Phase-contrast microscopy was used to observe the cell morphologies.

RESULTSThe cytokine group exhibited higher proportions of typical immature CD8a(+) DC, especially on day 3, but the total cell number and DC2 proportion decreased during prolonged culture. The low GM-CSF control group showed the same tendencies as the cytokine group on days 16 and 22, but produced higher total cell numbers (P<0.05) with lower DC2 proportions and cell numbers. The control group without exogenous cytokines spontaneously generated a certain proportion of DC2, but with low total cell and DC2 numbers that decreased rapidly, especially during prolonged culture (days 7 and 16, P<0.05).

CONCLUSIONSCulture in the presence of 5 ng/ml GM-CSF, 25 ng/ml Flt3L, 20 ng/ml IL-4 and 100 ng/ml SCF can rapidly induce large quantities of predominant immature CD8a(+) DC from murine bone marrow cells. Therefore, these represent optimal culture conditions for preparing murine immature DC2 in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; CD8 Antigens ; metabolism ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; Cell Culture Techniques ; methods ; Cells, Cultured ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Microscopy, Phase-Contrast