Objective To investigate the role of Keratin 13 gene in laryngeal carcinogenesis. Methods To detect the deletion frequency of keratin 13 sequence indirectly by LOH analysis at DNA level using 5 STR primers within and near keratin 13 gene in 100 cases. Results LOH was found in all of the microsatellite loci, and the LOH frequencies were 30.48%, 26.02%, 21.62%, 37.66% and 21.51% at D17S1964E, D17S2092, D17S791, D17S1665, and D17S808 positions respectively. The frequencies of LOH were not related to the type of laryngeal carcinoma. Conclusion Keratin 13 gene might play an important role in the laryngeal carcinogenesis,and further research is necessary to confirm it.

Objective To discuss the expression and mutation of β-catenin gene in breast carcinoma.Methods 119 breast carcinomas, 72 adenosis of breast tissues and 40 normal breast tissues were studied.Using immunohistochemistry studies to detect the expressions of β-catenin, then analyzing their relationship with the clinicopathological features of breast carcinomas. Gene mutation of β-catenin (exon 3) in 44 cases of breast carcinoma were analyzed by polymerase chain reaction (PCR) and then by direct sequencing.ResultsIn 40 cases of normal breast tissues, epithelial cells showed equally strong membranous of β-catenin protein at the cell-cell boundaries. The abnormal rate of β-catenin in breast carcinomas (78.15 %) was much higher than that of adenosis of breast tissues (44.44 %), the significant correlation was found (P＜0.01). The abnormal expression of β-catenin was associated with lymph node metastasis (P=0.031). Mutation of β-catenin gene was not detected in 44 cases of breast carcinoma, while the abnormal rate of β-catenin in 44 cases of breast carcinoma was 77.30 %. Conclusionβ-catenin abnormal expression could be considered as an useful marker for determining metastasis and prognosis of human breast carcinoma and for guiding treatment. β-catenin abnormal expression is not caused by β-catenin gene mutation, maybe by other mechanisms. It needs further study.

BACKGROUND:Insulin-like growth factor-1 (IGF-1) is an important mitogen and an indispensable regulator during normal hair cell development. Extracellular signal regulated kinase 1/2 (ERK1/2) is also expressed in mammalian vestibular organs.OBJECTIVE: To investigate the changes of the expression and the distribution of IGF-1, IGF-1 receptor (IGF-1R) and ERK1/2 in guinea pig's vestibular epithelium following gentamycin injury.DESIGN: A randomly controlled study.SETTING: Department of Otorhinolaryngology, General Hospital of Chengdu Military Area Command of Chinese PLA.MATERIALS: Twenty healthy and adult guinea pigs, weighing 300 to 350 g, provided by the Experimental Animal Center,Fourth Military Medical University of Chinese PLA, were randomly divided into four experimental groups and a control group with 4 in each group.METHODS: The experiment was performed at Department of Otorhinolaryngology Research Laboratory of Xijing Hospital,Fourth Military Medical University of Chinese PLA from January 2002 to May 2002. In the experimental groups,gentamycin was intraperitoneally administered daily (80 mg/kg) for 10 consecutive days. For the control group, 1 mL of 0.9% sterile saline was administered in the same way. The guinea pigs, in which unsteady gait or nystagmus appeared,could be involved in the further experiments. Before being sacrificed, the animals' gait, nystagmus and the amount of food-intake were observed every day. The animals were anesthetized deeply with 50 mg/kg pentobarbital 1, 7, 14 or 21days after the last gentamycin administration and then decapitated. Bilateral otic vesicles were harvested and immersed in 4% paraformaldehyde. Then the specimens were decalcified in 10% ethylenediamine tetraacetic acid (EDTA),thereafter embedded in OCT gel and serially sectioned at a thickness of 10 μm in the cryostat. The behavioral changes of guinea pigs were observed. The morphological changes of vestibular epithelium were examined by hematoxylin and eosin (HE) staining. The expression and distribution changes of IGF-1/1R and ERK1/2 were investigated immunohistochemically. MAIN OUTCOME MEASURES: The behavioral, morphological, expression and distribution changes of IGF-1/1R and ERK1/2 of guinea pigs in each group.RESULTS: Twenty involved guinea pigs entered the stage of final analysis. ① All the guinea pigs' vestibular functions were impaired after gentamycin treatment, then partially restored without any pharmaceutical treatment. ②Morphological injury was obvious after gentamycin treatment, and improved 1 week later. ③The expression of IGF-1/1R was low in the control group, and in the 1-day group it increased significantly to its maximum. After that, its expression decreased gradually, but was still higher in the 21-day group than that in the control group. There were significant differences in the expression of IGF-1/1R among different groups (F =51.8,45.7,P ＜ 0.05). The expression changes of IGF-1 and its receptor were similar. ④The expression of ERK1/2 was low in the control group, and increased gradually after gentamycin toxicity. In the 7-day group, the immunoreactivity reached its maximum. Then the levels of ERK1/2 decreased gradually, but were still higher in the 21-day group than that in the control group. There were significant differences in the expression of ERK1/2 among different groups (F =103.7,106.4, P ＜ 0.01), but no significant differences existed in the expression between ERK1 and ERK2 among different groups (P ＞ 0.05).CONCLUSION: The expression of IGF-1/1R and ERK1/2 in vestibular epithelium increased after gentamycin treatment,and IGF-1 may play an important role as an endogenous mitogen through a paracrine or autocrine manner in the early stage of hair cell self-repair of guinea pigs. ERK1/2 may play an important role in signal transduction during vestibular hair cell self-repair after gentamycin toxicity in guinea pigs.

Objective an experimental animal model of acute myoc ardial infarction (ANI) was established by opening chest and ligation of the left anterior descending (LAD) coronary artery． Methods a total of 20 rabbits were opened chest and ligated LAD under sterilization． Electrocardiogram (ECG) and myocardial-enzymes in blood serum were investigated． Results ECG of all rabbi t s showed normal before operation． Irmediately after and 1/2 hour after ligation , ST-segment elevated and ECG showed ambulatory changes for 7 and 9 rabbits respectively． Two hours after LAD ligation, the change of ECG for 2 rabbits wa s not typical and 2 of them died during experiment． Four weeks after operation, E CG of 18 rabbits showed the chest leads had pathologic Q waves． Twenty-four ho urs after LAD ligation, AST, LDH, LDH-1, CK and CK-MB in blood serum were significantly increased． There was significant difference compared with before operation (except LDH) (P<0. 0l)． Conclusions:The method was sim ple and well repeated． The formation of myocardial infarction was reliable and rabbits were maintained for a long time after operation． It provides a valuable animal mode l for the experiment study of coronary heart disease．

ObjectiveTo explore the effect of earlier rehabilitation on activities of daily living(ADL) of patients with spinal cord injury(SCI).Methods50 SCI patients received earlier rehabilitation and improvement of ADL of patients was evaluated.ResultsAfter two months treatment,the scores of Barthel index,functional independence measure(FIM) grade of patients increased significantly compared with that of before treatment(P<0.05) and ADL improved.ConclusionEarlier rehabilitation can improve ADL of SCI patients.

Humans ; Metals ; Microwaves ; adverse effects ; Occupational Exposure ; adverse effects ; analysis ; Workplace

Objective To evaluate the protective effect of Bletilla striata polysaccharide ( BSPS) on immunological and chemical liver injury in mice. Methods Thirty Kunming male mice were randomly divided into five groups, including the normal control group,model control group,and low-,middle-,and high-dose BSPS groups (n=6 each).Tail vein injection of ConA was carried out to establish the ConA-induced liver injury model.After different treatments,all the animals were sacrificed,and the plasma levels of ALT and AST were tested.Additionally,sixty Kunming male mice were randomly divided into six groups,including the normal control group,model control group,silymarin group,and low-,middle-,and high-dose BSPS groups (n=10 each).Tail vein injection of CCl4 was performed to establish the CCl4-induced acute liver injury model.After different treatments,the plasma levels of ALT and GSH were tested.The effects of BSPS on the weights of the liver and spleen were examined. Results The levels of ALT and AST were reduced in BSPS-treated mice when compared with those experiencing only ConA-induced liver injury ( model control group) ,and significant difference was found between the middle-and high-dose BSPS groups and the model control group (P<0.01,P<0.05).The weights of the liver and spleen and the level of ALT were reduced in BSPS-treated mice as compared with those with only CCl4-induced acute liver injury (model control group),while the level of GSH was significantly increased in middle-and high-dose BSPS groups (P<0.05). Conclusion BSPS at low,middle,and high doses can prevent against the ConA-induced immunological liver injury and CCl4-induced acute liver injury in mice.

The method of multiplex PCR was set up to identify two or three transgenes in one reaction such as uidA and bar; uidA and IDx5 or uidA, bar and 1Dx5 genes. Three sets of primer pairs which was specific to each of these three genes respectively were designed and synthesized. Recombinant plasmids pAHC25 and p1Dx5 harboring uidA + bar and 1Dx5 gene separately were used as template DNA in the process of optimizing an multiplex PCR reaction. The optimal annealing temperature for uidA and bar MPCR is range from 57. 1℃-62. 3℃ , for uidA and 1Dx5 is range from 60℃ to 60. 6℃ , and for uidA、bar and 1Dx5 range from 57. 0℃-58. 4℃. The amount of template for MPCR is twice as much as that for simplex PCR, while the concentration of primers is the same with simplex one. Less than 50bp MPCR products can be separated clearly by 10% non-denaturalized polyacrylamid gel electrophoresis. Fourteen transgenic wheat lines were tested by multiplex and simplex PCR respectively, which shows the same results and hence presents that MPCR is the reliable, rapid and high-effective approach to detect foreign genes from transgenic plant.

Objective To measure the expressions of cytotoxity-related cytokines in CD8+memory T cell subsets of peripheral blood in patients with multiple sclerosis (MS) and further analyze the relationship between the cytokine levels and MS disease severity. Methods Four-color flow cytometry was employed to detect the proportion of CD8+ memory T cell subsets expressed perforin and granzyme-B in peripheral blood of patients with MS, with other neurological disorders (OND) and of normal control (NC) group. The expanded disability status scale (EDSS) was used to reflect the disease severity at the time of blood sampling. Results Patients with MS had a decreased proportion of effector memory CD8+ T (CD8+ TEM) cells and terminal TEM cells expressed granzyme-B as compared with NC group (P＜0.05). CD8+ TEM cells expressed granzyme-B and perforin were negatively correlated to the scores of EDSS in patients with MS (r=-0.493, P=0.027; r=-0.594, P=0.009). Conclusions CD8+ TEM may be involved in the MS-related CNS inflammatory immune response. The proportion of CD8+ TEM cells expressed perforin and granzyme-B can be used to reflect the pathological severity of MS lesions to some degrees.