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Objective: To investigate the effect of ubiquitin mutation at position 331 of tumor necrosis factor receptor related factor 6 (TRAF6) on the biological characteristics of colorectal cancer cells and its mechanism. Methods: lentivirus wild type (pCDH-3×FLAG-TRAF6) and mutation (pCDH-3×FLAG-TRAF6-331mut) of TRAF6 gene expression plasmid with green fluorescent protein tag were used to infect colorectal cancer cells SW480 and HCT116, respectively. The infection was observed by fluorescence microscope, and the expressions of TRAF6 and TRAF6-331mut in cells was detected by western blot. Cell counting kit-8 (CCK-8) and plate cloning test were used to detect the proliferation ability of colorectal cancer cells in TRAF6 group and TRAF6-331mut group, cell scratch test to detect cell migration, Transwell chamber test to detect cell migration and invasion, immunoprecipitation to detect the ubiquitination of TRAF6 and TRAF6-331mut with ubiquitinof lysine binding sites K48 and K63. Western blot was used to detect the effects of TRAF6 and TRAF6-331mut over expression on the nuclear factor kappa-B (NF-κB) and mitogen activated protein kinase mitogen-activated protein kinase (MAPK)/activating protein-1(AP-1) signal pathway. Results: The successful infection of colorectal cancer cells was observed under fluorescence microscope. Western blot detection showed that TRAF6 and TRAF6-331mut were successfully expressed in colorectal cancer cells. The results of CCK-8 assay showed that on the fourth day, the absorbance values of HCT116 and SW480 cells in TRAF6-331mut group were 1.89±0.39 and 1.88±0.24 respectively, which were lower than those in TRAF6 group (2.09±0.12 and 2.17±0.45, P =0.036 and P =0.011, respectively). The results of plate colony formation assay showed that the number of clones of HCT116 and SW480 cells in TRAF6-331mut group was 120±14 and 85±14 respectively, which was lower than those in TRAF6 group (190±21 and 125±13, P =0.001 and P =0.002, respectively). The results of cell scratch test showed that after 48 hours, the percentage of wound healing distance of HCT116 and SW480 cells in TRAF6-331mut group was (31±12)% and (33±14)%, respectively, which was lower than those in TRAF6 group [(43±13)% and (43±7)%, P =0.005 and 0.009, respectively]. The results of Transwell migration assay showed that the migration numbers of HCT116 and SW480 cells in TRAF6-331mut group were significantly lower than those in TRAF6 group ( P <0.001 and P <0.002, respectively). The results of Transwell invasion assay showed that the number of membrane penetration of HCT116 and SW480 cells in TRAF6-331mut group was significantly lower than those in TRAF6 group ( P =0.008 and P =0.009, respectively). The results of immunoprecipitation detection showed that the ubiquitin protein of K48 chain pulled by TRAF6-331mut was lower than that of wild type TRAF6 in 293T cells co-transfected with K48 (0.57±0.19), and the ubiquitin protein of K63 chain pulled down by TRAF6-331mut in 293T cells co-transfected with K63 was lower than that of wild type TRAF6 (0.89±0.08, P <0.001). Western blot assay showed that the protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-HCT116 cells were 0.63±0.08, 0.42±0.08 and 0.60±0.07 respectively, which were lower than those in TRAF6-HCT116 cells ( P =0.002, P <0.001 and P <0.001, respectively). The expression level of AP-1 protein in TRAF6-HCT116 cells was 0.89±0.06, compared with that in TRAF6-HCT116 cells. The difference was not statistically significant ( P >0.05). The protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-SW480 cells were 0.50±0.06, 0.51±0.04, 0.48±0.02, respectively, which were lower than those in TRAF6-SW480 cells (all P <0.001). There was no significant difference in AP-1 protein expression between TRAF6-331mut-SW480 cells and TRAF6-SW480 cells. Conclusion: The ubiquitin site mutation of TRAF6 gene at 331 may prevent the binding of TRAF6 and ubiquitin lysine sites K48 and K63, and then affect the expressions of proteins related to downstream NF-κB and MAPK/AP-1 signal pathways, and inhibit the proliferation, migration and invasion of colorectal cancer cells.

Objective: To find the drugs for lowering the pulmonary vascular resistance in patients of chronic obstructive pulmonary disease (COPD) but would not increase its side-effect., Methods: Changes of some hemodynamic parameters in 15 patients of COPD in stable stage treated by nitrendipine and ligustrazine were observed by right heart floating catheterization., Results: The mean pulmonary arterial pressure reduced by 1.4% immediately after the combined treatment, and reduced by 17.0%, 20.0% and 18.0% in 15 mins, 30 mins and 60 mins after treatment. In the responding time, the pulmonary vascular resistance reduced by 15.2%, 36.2%, 43.0% and 34.6%, the systemic vascular resistance reduced by 7.9%, 19.2%, 17.9% and 20.8%, and the cardiac output increased by 15.8%, 22.6%, 22.2%, and 33.8% respectively. The changes of systemic arterial pressure and heart rate were of no statistical significance., Conclusion: Combination therapy of nitrendipine and ligustrazine could lower the pulmonary arterial pressure and pulmonary vascular resistance effectively and has a selective effect on pulmonary circulation in patients with COPD.

Objective: To study the therapeutic mechanism of combination therapy of ligustrazine and nitrendipine in treating patients with chronic obstructive pulmonary disease (COPD)., Methods: Thirty COPD patients divided in to 3 groups (10 in each) were treated with ligustrazine, nitrendipine and ligustrazine plus nitrendipine respectively, and the changes of hemorrheologic parameters, plasma endothelin (ET-1), thromboxane A2(TXA2) and platelet-P-selectin (CD62P) before and after treatment were observed., Results: The combination therapy of ligustrazine and nitrendipine could lower the levels of plasma ET-1, TXA2, CD62P and the hemorrheologic parameters., Conclusion: Combination of ligustrazine and nitrendipine showed a therapeutic effect better than that of the two drugs used separately. Its effect in lowering pulmonary circulation resistance is related with the lowering of plasma vaso-contrictive factor and the changing of hemorrheologic properties. The integrated traditional Chinese and western medical therapy is valuable in treating COPD.

We reviewed the epidemiologic and clinical features of ruptured intracranial aneurysms based on the available data in the literature reported in Taiwan. Among 28,763 stroke patients registered by members of the Neurological Society of Taiwan, 600 patients had verified intracranial aneurysms (2.1%), constituting only 34.7% of 1,730 patients with subarachnoid hemorrhage. The male to female ratio was 0.68.1. The mean age was 54.6 +/ 14.3 years in men, and 56.5 +/ 13.8 years in women. Among the 409 patients whose aneurysmal sites were recorded, the aneurysm occurred at the anterior communicating artery in 109 (26.7%), at the posterior communicating artery in 90 (22.0%), at the internal carotid artery in 82 (20.1%), at the middle cerebral artery in 80 (19.6%), at the anterior cerebral artery in 50 (12.2%), at the vertebral-basilar system in 13 (3.2%), and multiple sites in 33 (8.1%). Cerebral angiography was performed in 72.2% of patients and operation was undergone in 57.7%. A randomized follow-up study in 59 patients showed survival rates of 63% for 1 month, 53.3% for 6 months and 43.6% for 1 year. The reported surgical mortality rates ranged from 7.7% to 12.5%. Prognosis was related to the score on the World Federation of Neurological Societies Subarachnoid Hemorrhage Scale. In Taiwan, the number of patients with intracranial aneurysm has been underestimated among who have presented with subarachnoid hemorrhage. The high mortality and morbidity of aneurysm rupture justify an aggressive attempt to establish a prompt and accurate diagnosis.

In order to study the effect of dihydroxyacetophenone (DHAP) on pulmonary hemodynamics and its relationship to plasma ANP as well as cAMP/cGMP level in chronic obstructive pulmonary disease (COPD), 11 COPD patients were examined with the right heart catheterisation, and the plasma ANP and cAMP/cGMP were measured with radioimmunoassay at the same time. The results showed that intravenous given DHAP 640 mg could decrease mean pulmonary arterial pressure, pulmonary vascular resistance and systemic vascular resistance (P < 0.05), but increase the cardiac output from 4.10 +/- 1.08 L/min to 5.13 +/- 1.19 L/min (P > 0.05) and not affect systemic arterial pressure (P > 0.05) as well as blood gas analysis; it also reduce the plasma ANP and cGMP level from 0.73 +/- 0.42 pg/ml to 0.41 +/- 0.33 pg/ml (P < 0.01) and from 9.82 +/- 5.75 pm/ml to 8.01 +/- 4.80 pm/ml (P < 0.05) respectively, but did not affect the plasma cAMP level (P > 0.05). It is suggested that DHAP may relax pulmonary vessels by regulating the ratio of cAMP to cGMP, and the lowering of plasma ANP level might be a secondary reaction. So we consider that DHAP is a fast-acting, safe and potential drug in treating pulmonary arterial hypertension by COPD.

Two cases brothers of Peutz-Jeghers syndrome are reported, in which the gastrointestinal polyps were resected by laparotomy and enterotomy. One of the polyps was in the cecum and was found to have atypia cellular change, though no malignancy was found in the others, which were thought to be hamartomas. The hamartomatous polyps were initially thought to have little potential for malignancy, and the disease was believed to have a relatively benign course. However, reports in the literature have associated this syndrome with various kinds of malignancies, and the overall risk of cancer in this population is regarded as higher than normal. The analysis of our cases and a review of the literatures are presented for clarification of the predisposition of the development of malignancies in these patients.

Giant cell reparative granuloma has been described mainly involving the mandible and is very uncommon in the temporal bone. It is not a true neoplasm, but actually represents a local reparative reaction to hemorrhage, usually following trauma. The treatment of choice is surgical curettage of the lesion. The prognosis is usually good. Radiation therapy with small dose may be considered if the lesion is not amenable to surgical approach. This paper describe a giant cell reparative Granuloma which occurred in the temporal bone of a 41 year-old man. The patient accepted surgical treatment in 1983 but the granuloma subsequently recurred in 1986. The radiation therapy was then given approximately 5000 rads. No sign of recurrence has been found up to the time of this writing.

57 specimens of raw foods and 143 specimens of ready to eat foods were examined for microbial contamination in a period from October, 1982 to September, 1983. The results showed that 26% of the raw foods and 24% of the ready to eat foods didn't meet the government's requirement. When TAPC less than 10(6) CFU/g is used as acceptable level, then 60% of the raw foods and 46% of the ready to eat foods are unsatisfactory. The appearance of pathogenic bacteria in foods increased when the TAPC rised, although only the emergence of S. aureus was statistically significant. The number of indicator bacteria in the foods examined didn't show any correlation with the existence of pathogenic bacteria, indicating that the presence of coliform bacteria and E. coli in foods could hardly serve as parameters for evaluating food safety.