Objective To study the time course of effect of methylene blue on inducible nitric oxide synthase (iNOS) mRNA transcription and protein expression in lung tissue of rats with sepsis, and its mechanism. Methods 126 female Wistar rats were randomly divided into sham group, sepsis group and methylene blue group. Each group was subdivided into 0-, 6-, 12-, 18-, 24-, 30-, and 36-hour subgroups according to the time after operation, with 6 rats in each subgroup. A model of sepsis was reproduced by cecal ligation and puncture (CLP), and the rats in sham group were only opened the abdominal cavity and isolated the membrane of the appendix without CLP. Rats in methylene blue group were given injection of 15 mg/kg methylene blue at all time points after CLP, the remaining rats were given 0.9%NaCl solution in same amount. Six hours after the injection, the rats were sacrificed and the lung tissue was harvested immediately. The expression of iNOS mRNA and protein in lung tissues were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and Western Blot respectively, and the changes in histopathology were observed using hematoxylin and eosin (HE) staining. Results Compared with sham group, the expression of iNOS mRNA was significantly up-regulated at 6, 12, 18 and 24 hours after CLP in sepsis group (2-ΔΔCt: 2.42±0.66 vs. 1.00±0.38 at 6 hours, P = 0.002; 2.54±0.76 vs. 1.00±0.27 at 12 hours, P = 0.000; 5.46±2.26 vs. 1.00±0.38 at 18 hours, P = 0.000; 3.03±0.62 vs. 1.00±0.33 at 24 hours, P = 0.001), and iNOS protein expression was significantly up-regulated at 12, 18 and 24 hours (gray value: 2.54±0.45 vs. 1.00±0.35 at 12 hours, P = 0.000; 2.65±0.64 vs. 1.00±0.33 at 18 hours, P = 0.000; 3.03±0.59 vs. 1.00±0.24 at 24 hours, P = 0.000). Compared with sepsis group, the expression of iNOS mRNA was significantly down-regulated at 6, 12, 18 and 24 hours in methylene blue group (2-ΔΔCt: 1.55±0.82 vs. 2.42±0.66 at 6 hours, P = 0.034; 1.84±0.42 vs. 2.54±0.76 at 12 hours, P = 0.016; 2.66±1.09 vs. 5.46±2.26 at 18 hours, P = 0.003; 2.20±0.29 vs. 3.03±0.62 at 24 hours, P = 0.002), and iNOS protein expression was significantly lowered at 12, 18 and 24 hours (gray value: 1.84±0.18 vs. 2.54±0.45 at 12 hours, P = 0.003; 1.87±0.27 vs. 2.65±0.64 at 18 hours, P = 0.008; 2.20±0.50 vs. 3.03±0.59 at 24 hours, P = 0.008). Histopathological observation showed that the degree of lung injury at each time point, including red blood cells effusion, lung interstitial edema, inflammatory cell infiltration, alveolar collapse etc., in sepsis group and methylene blue group were significantly higher than that of sham group, and the degree of lung injury in rats with methylene blue was not significantly improved as compared with that of sepsis group. Conclusions Lung iNOS mRNA expression was significantly increased at 6-24 hours after CLP induced sepsis in rat, and protein expression was increased at 12-24 hours. Methylene blue could inhibit mRNA transcription and protein expression of iNOS in lung of septic rat, but failed to reduce the degree of lung injury in sepsis.

Objective To study the effectiveness of the complete ureterel reconstruction using bladder flap after cadaveric renal transplantaion.Methods In 13 recipients with complete necrosis of ureter after renal transplant from July 1995 to October 2003, tubelike bladder flaps were applied to ureteraplasty substitute for the necrosis ureter. Artificial pyeloureterostomy was performed and double T tubes were used as the stents. Routine drainage-tubes were placed in incisions. Results All patients were successful in ureteral reconstruction. The renal function of 10 patients was improved significantly 4 weeks postoperation. Nephrectomy was performed in one patient because of severe circumrenal infection 7 days after operation. One-year survival rate of recipients and allografts was ~100 % (13/13) and ~92.3 % (10/13) respectively. Urine reflow occurred in 2 patients during a follow-up of 12 months. Conclusion The use of bladder flap for ureteral reconstruction is an effective technique for total autograft ureteral necrosis.

Objective To compare the immunostaining of human epidermal growth factor receptor-2(HER-2) among three different clones,and to compare with the results of fluorescence in situ hybridization(FISH) detection,to provide evidences for pathological department to select appropriate antibodies.Methods 268 cases of invasive breast cancer were examined by immunohistochemistry using antibodies for HER-2 of 3 different clones.The results were compared with those done by FISH.Results Immunostaining using antibody clone SP3 showed 3+ reactions in 66 cases;Immunostaining using antibody clone EP3 showed 3+ reactions in 53 cases,while using polyclonal HER-2 antibody,80 cases showed 3+ reactions.There was statistical significance between those groups using different clones(P<0.05).Consistent with the FISH results,the Kappa values are 0.76,0.67,and 0.56,respectively.Conclusion It suggests detection of HER-2 using immunohistochemistry should prioritize selecting monoclonal antibodies,especially clone SP3.

Objective To determine the effect and safety of metallic stents in the treatment of acute malignant colorectal obstruction.Methods From May,2000 to June,2003, self-expanding metallic stents were implanted in 15 patients with acute left colonic obstruction caused by malignancies. The postoperative remission and complications were observed.Results Stents were implanted successfully in 12 cases(80.0%,12/15).Of the 12 cases,the obstruction were all disappeared within 24h. No death or colonic perforation happened in this series. Stent migration and anal pain developed each in one case after stent implantation. Elective radical resection was performed on 8 cases after bowel preparation and supportive therapy; and stents reserved permanently in other 4 cases. Conclusions The implantation of self-expanding metallic stents is a safe and effective method for the temporary remission or permanent treatment of acute left-colonic or rectal obstruction caused by malignancies,it can instead of colostomy.

Objective To investigate clinical result of hepatectomy and porta-enterostomy in the treatment of Bismuth type Ⅳ hilar cholangiocarcinoma.Methods Nine patients with Bismuth type Ⅳ hilar cholangiocarcinoma underwent accurate hilar resection(portal parecnchyma resection including about Ⅰ cm of the hilar part of the segments 5 and 4b and caudate lobe beyond the tumor),and the biliary drainage was reconstructed by Roux-en-Y portal parecnchyma-jejunum lpop anastomosis.None of the biliary radicals had to be ligated and all of them were drained into thus constructed"biliary pool".Results Hilar resection was successfully performed in all cases,and there was no postoperative mortality.Aspartate transaminase and alanine transaminase and serum bilirubin decreased evidently four weeks later.Three patients presented postoperative complications.One patient developed a transient anastomotic leakage,while one patient developed self-limiting hemobilia,wound infection occurred in one patient.All three patients were treated conservatively and recovered.The mean Karnofsky performance score was 86,with which they could carry on normal activity with minor symptoms of disease.Two patient died after 9 months and 17 months of extensive metastasis and intrahepatic metastasis respectively.The remaining seven patients are alive by a mean followup of 24.9 months after surgery without any signs of recurrence.Conclusions With accurate hilar resection and portal parecnchyma-to-enterostomy,the patients considerably benefit from the preservation of liver parenchyma and patent biliary drainage and radical resection.So the Hew technique prolongs the survival time and enhances the quality of life of the patients.

Objective To explore the relationship between plasma homocysteine levels and diabetic retinopathy (DR).MethodIn 40 DR patients and 63 non-DR patients,the indexes including plasma homocysteine concentrations,duration of diabetes,ages,blood pressure,preprandial blood glucose(PBG),plasma GHbA,e,plasma lipids,serum folie acid and semm vitamin B12 were analyzed.ResultsPlasma homocysteine level was significantly higher in DR patients (12.9μmol/L) than that in non-DR patients (7.8μ.mol/L) (P＜0.01),and the relationship remained significant correlation after adjusted for duration of diabetes,GHbA1c,ages,serum folic acid and serum vitamin B12(OR = 1.23,P＜0.05).ConclusionHyperhomocysteinimia is a risk factor and an independent predieator of DR.

Objective To observe the effect of methylene blue on the expression of liver inducible nitric oxide synthase (iNOS) in rats with different stages of sepsis.Methods One hundred and twenty-six adult female Wistar rats were randomly divided into three groups: sham operation group, sepsis group and methylene blue group, each group was again subdivided into 0, 6, 12, 18, 24, 30, 36 hours subgroups, each subgroup6 rats. The model of sepsis was established by cecal ligation and puncture (CLP) method, and in the sham operation group, the abdominal incision was performed and the intestinal mesentery was separated only, without ligation and perforation. In methylene blue group, 15 mg/kg methylene blue was injected into a caudal vein at 0, 6, 12, 18, 24, 30, 36 hours after CLP in the rats in corresponding subgroups, respectively; in the sepsis and sham operation subgroups, the same amount of 0.9% normal saline was given. After administration for 6 hours in various groups, the rats were sacrificed and the liver tissue was harvested immediately. The expression of iNOS mRNA of liver tissues was determined by the real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR),and the protein expression of iNOS was determined by Western Blot.Results Compared with sham operation group, the liver tissue expression of iNOS mRNA was significantly up-regulated in sub-sepsis groups at 0, 6, 12 and 18 hours after CLP (2-ΔΔCt: 16.66±2.81 vs. 1.00±0.36, 12.26±5.78 vs. 1.00±0.30, 6.08±1.33 vs. 1.00±0.18, 2.42±0.64 vs. 1.00±0.12, allP < 0.01), after 24 hours the expression of iNOS mRNA had no significant change; the liver tissue expression of iNOS protein was obviously up-regulated in sub-sepsis groups at 6, 12 ,18 and 36 hours after CLP (gray value: 0.350±0.011 vs. 0.210±0.005, 1.460±0.085 vs. 0.090±0.005, 0.230±0.012 vs. 0.18±0.008, 0.310±0.017 vs. 0.200±0.010, allP < 0.01). Compared with sepsis group, the expression of the liver tissue iNOS mRNA was down-regulated in methylene blue subgroups at 0, 12 and 18 hours after CLP (2-ΔΔCt: 9.90±3.06 vs. 16.66±2.81, 1.56±0.58 vs. 6.08±1.33, 1.11±0.15 vs. 2.42±0.64, allP < 0.05), and the expression of iNOS protein was down regulated in methylene blue subgroups at 6, 12, 18 and 36 hours after CLP (gray value: 0.150±0.008 vs. 0.350±0.011, 0.950±0.009 vs. 1.460±0.085, 0.150±0.007 vs. 0.230±0.012, 0.170±0.009 vs. 0.310±0.017, allP < 0.05).Conclusion Zero-24 hours after CLP, the expressions of mRNA iNOS and protein in liver of septic rats are significantly increased; methylene blue can markedly inhibit the expressions of iNOS mRNA and protein in the liver of rats with sepsis.

Objective To investigate the molecular characteristics and epidemiological signification of patients with low-level HBsAg. Methods PCR and gene sequencing were used to detect HBV DNA and Tyr-Met-Asp-Asp(YMDD) mutant in 136 serum samples with low-level HBsAg and 44 sernm samples with high-level HBsAg. Genotyping was performed in 47 cases with HBV DNA 10~5 copies/L by concentration method and 37 cases with high-level HBsAg. S gene sequences and serotypes were analyzed in 14 cases with HBV DNA 105 copies/L and 29 cases with high-level HBsAg. S gene sequences were compared with the consensus sequence of Chinese strain by BioEdit software. Results The HBV DNA-positive rate, YMDD mutation rate and HBV DNA load (logarithm) in low-level and high-level HBsAg group were 34.6% (47/136), 0% (0/136), 6.5±1.4 and 84.1% (37/44), 9.1% (4/44), 8.9±1.8, respectively. There was statistically significant differences between two groups (for concentration method,χ~2 = 30.8, P < 0.05; for direct method, χ~2 = 53.5, P < 0.05; for YMDD mutation ratio, P = 0.003, For HBV DNA (log), t = 6.5, P < 0.05). The genotypes in low-level HBsAg group included type B (16/47), type C (5/47) and non-classified ones(26/47). There were significant differences between two groups (χ~2=21.8, P <0.01). The serotypss included adw (7/14), ayw (4/14), adr (2/14) and ayr (1/14). There were significant differences in genotypes (χ~2 = 13.5, P < 0.05) but not in serotypes between two groups (χ~2 = 4.7, P >0.05). S gene sequencing results showed no S gnne variation was detected, but there were 6 single nucleotide polymorphisms in 16 cases, which would not result in the alternation of amino acid. Conclusions Low-replication phenomenon of HBV DNA was present in patients with low-level HBsAg. The major genotyps and serotype was type B and adw/ayw, respectively. Polymorphic variants have been found in the S gene. The existence of low-level HBsAg might be related with its own molecular characteristics resulting in low expression of HBsAg or immune tolerance induced by low-level HBsAg after HBV infection.

OBJECTIVETo study the effects of rare earth exposure on human telomerase and apoptosis of human peripheral mononuclear cells (PBMNs).

METHODSRare earth mine lot in Xunwu county, the biggest ion absorptive rare earth mine lot of China, was selected as the study site. Another village of Xunwu county, with comparable geological structure and social environment was selected as the control site. Thirty healthy adults were randomly selected from the study site as exposure group and another 30 healthy adults randomly selected from the control site as control group. The blood content of 15 rare earth elements, including La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu and Y, were determined by inductive coupled plasma-source mass spectrometry (ICP-MS). The total contents of rare earth elements in the blood were calculated. The TRAP and FCM assays were carried out to analyse the telomerase and apoptosis of human PBMNCs respectively.

RESULTSIn the exposure group, the concentration of La, Ce, Dy and Y were significantly higher (P<0.001), and Pr, Nd, Sm, Gd and Yb were higher than those in the control group (P<0.05). The total content of rare earth in the blood of exposure group showed significant difference compared with control group (P<0.001). Telomerase activity in PBMNs of the exposure group was higher than that in the control group (P<0.05); there were 11 adults in the exposure group (30 adults) and 5 adults in control group (30 adults) showed positive telomerase activity. The average age of the exposure group was (38.69 +/- 8.02) years-old, while the control group was (40.45 +/- 9.02) years-old (P >0.05). It was found that there was a significant relationship between telomerase activity and the total content of rare earth elements (P <0.01). 3. The proportion of apoptosis was not different between the two groups (P >0.05), but the cells in the S-phase and G2-M phase were increased (P <0.01) in the exposed group.

CONCLUSIONThe telomerase activity of PBMNs in the rare earth elements exposed group was higher than that of the control group, and there is no effect on apoptotic rate of PBMNs, but may promote the diploid DNA replication, and increase the percentage of G2/M and S phase cells.

Apoptosis ; drug effects ; Environmental Exposure ; adverse effects ; Female ; Humans ; Leukocytes, Mononuclear ; cytology ; enzymology ; Male ; Metals, Rare Earth ; adverse effects ; analysis ; Telomerase ; metabolism

This paper presents our work on the research process of virtual articulator simulation. Firstly, plaster models of upper jaw and low jaw are measured by 3D-optical measuring system and processed to get the available data. Secondly, complex physical modeling of the jaw motion is simplified and decomposed to a series of position transform, such as translation, rotation, etc.. Thirdly, opening/closing movement and laterotrusion is simulated by real-time updating display. Fourthly, the efficient collision detection algorithm is applied to dynamically compute occlusal contact regions and cutaway view. Finally, the current shortage of the virtual articulator simulation is discussed in detail and research prospect is forecasted.
Computer Simulation ; Dental Articulators ; Dental Occlusion ; Humans ; Imaging, Three-Dimensional ; Jaw ; physiology ; Models, Biological