Objective To study the expression of proliferating cell nuclear antigen (PCNA) in the occurrence and development of hepatocellular carcinoma. Methods Sixty SD rats were randomly divided into control group and experimental group. 3′-Me-DAB was administrated into rats to establish the experimental model of hepatocarcinoma. The expressions of PCNA of different phases were detected by immunohistochemistry and the liver pathologic changes were observed by optical microscope. Results The process of canceration was divided into three stages: inflammation, proliferative fibrosis and hepatic carcinoma. The expression of PCNA firstly presented in the oval cells that located in the portal area at the stage of inflammation, and a part of PCNA were hyper-expressed in the portal area. The expression rate of PCNA in the middle phase of inflammatory stage was higher than that of any other phases but declined later. Yet, when it came to the stage of hepatic carcinoma, the rate increased again. Conclusion Under the experimental circumstance when liver cancer is caused by the carcinogenic agent, PCNA may be firstly expressed in the oval cells, and the dynamic expression of PCNA may be an indicator for the early diagnosis of hepatocarcinogenesis.

Objective To explore the distribution characteristics of intra/extracranial large-artery disturbance in patients with ischemic stroke (IS)/transient ischemic attack (TIA).Methods The medical history information of 169 IS/TIA patients was recorded.The results of laboratory examination,electrocardiogram,echocardiography,abdominal ultrasound and chest radiography were analyzed.At the same time,the intra/extra-cranial large arteries were examined to analyze the degree,distribution and pattern of their disturbance.Results Among them,there were intra/extra-cranial large-arterial stenosis (n =112,66.3％) and small-arterial disturbance (n =57,33.7％).Multiple vascular disturbance pattern was more common in patients with intra/extra-cranial large-arterial stenosis (n =71,63.4％).The incidence of intracranial large-arterial stenosis was significantly higher than that of extracranial large-arterial stenosis (53.8％ vs.35.5％,P ＜ 0.05).And middle cerebral artery was the most commonly affected artery,36 of those were culprit vessels accounting for 73.5％ (36/49).Conclusions The IS/TIA patients have a high incidence of intra/extra-cranial large-arterial stenosis.Multiple vascular disturbance is a more common pattern.The incidence of intracranial large-arterial stenosis is greater than that of extracranial and middle cerebral artery is the most common affected artery.

Objective: To observe the clinical efficacy of Tuina (Chinese therapeutic massage) plus umbilical therapy for senile functional constipation due to Qi deficiency of lung and spleen.Methods: A total of 60 patients who met the inclusion criteria were randomly divided into a control group and an observation group by the random number table method, with 30 cases in each group. The patients in the control group were given lactulose oral solution, while the patients in the observation group were treated with Tuina and umbilical therapy. After two weeks of treatment, the scores of the Wexner continence grading scale (WCGS) and patient assessment of constipation-quality of life questionnaire (PAC-QOL) were observed. The effective rates in the two groups were compared. Results: After two weeks of treatment, the total effective rate in the observation group was 90.0％, while that in the control group was 76.7％, presenting a significant difference between the two groups (P<0.05). There were no significant differences in the WCGS and PAC-QOL scores between the two groups before treatment (P>0.05). After two weeks of treatment, the WCGS and PAC-QOL scores of both groups decreased significantly compared with those before treatment (P<0.05), and the two scores in the observation group were lower than those in the control group (P<0.05). Conclusion: The efficacy of Tuina plus umbilical therapy for senile functional constipation is confirmed. This method can improve patients' quality of life. Its efficacy is better than that of taking lactulose oral solution.

Purpose:To study the changes of t-PA ?u-PA expression and fibrinolysis molecular mark ers in patients with gastric or intestinal cancer in order to elucidate their cl inical significance. Methods:The plasma levels of t-PA ?u-PA ?u-PAR and PAP were measured by ELISA . Gen e transcription of t-PA?u-PA mRNA were detected by Real-time RT-PCR. Results:The plasma levels of u-PA?u-PAR and PAP were elevated in gastric or intestina l cancer patients ,while u-PA was markedly elevated in patients with local infi ltration?lymph node involvement or distal metastasis. u-PA mRNA was higher and t-PA was lower in gastric or intestinal cancer compared to normal tissue. Conclusions:Hyperfibrinolysis was an important factor related with m etastasis potential of gastric or intestinal cancer. t-PA may be a character of well differentiated tissue.

Objective: To evaluate feasibility of informant questionnaire of the Community Screening Instrument for Dementia(CSID) and related factors in a sample of community elderly population, and to investigate prevalence of suspectable dementia in the sampled people.Method: A total of 2267 people aged 65 and above in one rural area and one urban area in Beijing were recruited and their informants were assessed by the CSID informant questionnaire.Validity of CSID were evaluated according to the computerized diagnosis of 10/66 project and ICD-10.Result:The split-half reliability of CSID informant questionnaire was 0.85 and the consistency was more than 0.8.When the cutoff point of CSID was 6 or more, sensitivity was 86.2% and specificity was 93.2%.At meantime, the prevalence of suspectable dementia was 9.1%.Logistic regression analysis showed that the information obtained from informant as a spouse and from informant who lived in urban area was correlated with positive diagnosis of dementia.Conclusion:CSID informant questionnaire is valid as a screening instrument of dementia.Information from spouse is useful to identify the change of cognitive function of the elderly.

Objective To determine the possibility of telomere seeding in human gastric cancer cell SGC7901 and the influence of telomere on the expression of luciferase reporter gene nearby and to explore the position effect of telomere in gastric cancer cell. Methods The TET-OFF regulatory system was prepared and then transfected to gastric cancer cell SGC7901 by Lipofectamine TM 2000. With doxycycline added, the activity of the fluorescence caused by the expression of the target gene (the luciferase gene) with TET-OFF regulatory system was analyzed. The luciferase reporter plasmid in which telomere fragment pBX was inserted and the control plasmid pBX-ntf (no teloemere fragment) were co-transfected to cell 7901 with pTET-OFF regulate plasmid respectively and the luciferase activity was determined with doxycycline's induction. Results With the inducing of Dox, the activity of the fluorescence caused by the expression of the luciferase gene was inhibited. Treated with deoxycyline, there was a 4-fold decrease in the activity of the fluorescence in the cells infected with pBX and pTET-OFF as compared with that in the cells infected with pBX-ntf and pTET-OFF. Conclusion With the help of Lipofect2000, the gastric cancer cell SGC7901 can be transfected by TET-OFF regulatory system successfully. Dox could repress luciferase gene expression. Telomere seeding in gastric cancer cell SGC7901 could decrease the expression of luciferase reporter gene.

Objective To study the effect of telomere seeding on the biological behaviors of gastric cancer cell line and its mechanism. Methods The plasmid pSXneo-1.6-T2AG3, containing telomere fragment, was prepared in large scale and transfected into gastric cancer cell line SGC-7901 by LipofectAMINTM 2000. G418 was used to scan the positive clones and PCR was preformed to verify the stable seeding of the exogenous telomere in cell. MTT aassay and flow cytometry were used to determine the growth and cycle distribution of cells. The expression of cell cycle-related genes was detected by RT-PCR. Results Both pSXneo-1.6-T2AG3 eukaryonic expressing vector be inserted with telomere TTAGGG fragments and control vector were transfected into gastric cancer cell line SGC-7901 with LipofectamineTM 2000 and gain clones through G418 selection respectively, which were named as SGC-7901-telo and SGC-7901-neo. The cells were examined on DNA level by PCR to determine the correction of transfection. Flow cytometric analysis displayed an accumulation in G1M and G2M phase of cell cycle and an decreased percentage in S phase and proliferative index. Expression of p21 mRNA in 7901-telo cell line was increased (P0.05). Conclusion The pSX-T2AG3-neo eukaryonic expressing vector be inserted with 1600bp telomere TTAGGG fragments was successfully transfected into gastric cancer cell line SGC-7901 with the help of LipofectamineTM 2000. The seeding decreased malignant phenotypes of gastric cancer cell line and up-regulated the expression of p21, but the seeding did not distinctly influence on caspase3 mRNA expression.

Objective To investigate the percentage of Th17 cells and FOXP3 concentration in peripheral blood of children with bronchial asthma and their clinical significance .Methods Thirty children with bronchial asthma and thirty healthy children as con-trol group were enrolled in the study .The percentage of Th17 and CD4+CD25+ Treg cells in the peripheral blood were determined by flow cytometry(FCM) .The mRNA expression of FOXP3 in the peripheral blood was determined by quantitative real-time PCR . The concentrations of IL-17 and IL-10 in plasma were measured by using ELISA .Results In children with bronchial asthma ,the proportions of Th17 cells in the peripheral blood and concentration of IL-17 in plasma increased ,while the expression of FOXP3 in the peripheral blood and concentration of IL-10 in plasma decreased .Conclusion The imbalance of Th17/FOXP3 may contribute to the proceeding of bronchial asthma in children .

Objective To investigate characteristics of TCM syndromes of pulmonary tuberculosis,and to explore new methods for its treatment. Methods TCM syndrome scale was designed to investigate and summarize the characteristic of the syndrome, the syndrome of the general information, and the main clinical symptoms of 516 pulmonary tuberculosis patients. Results The between-group comparison of distribution of TCM syndromes in patients with pulmonary tuberculosis showed statistical difference (P＜0.05) . The occurrence rate of TCM syndromes in the 516 pulmonary tuberculosis patients were listed in descending order as follows:syndrome of both Qi and yin deficiency complicated with stasis ＞ syndrome of lung yin deficiency complicated with stasis＞syndrome of both Qi and yin deficiency＞syndrome of yin deficiency with various with age (P＜0.05) . Patients with blood stasis syndrome occupied 66.15% of the total number in the youth group; Patients with blood stasis syndrome occupied 78.03% of the total number in the middle-aged group;distribution of TCM syndromes was statistically different with the initial treatment and re-treatment (P＜0.05) .Patients with the blood stasis syndrome occupied 72.73% of the total number in the initial group, patients with effulgent fire ＞ lung yin deficiency syndrome.① The distribution of TCM syndromes was statistically various with age (P＜0.05) . Patients with blood stasis syndrome occupied 66.15％ of the total number in the youth group; Patients with blood stasis syndrome occupied 78.03％ of the total number in the middle-aged group; Patients with blood stasis syndrome occupied 78.22％ of the total number in the old-aged group. ② The distribution of TCM syndromes was statistically different with the initial treatment and re-treatment (P＜0.05) . Patients with the blood stasis syndrome occupied 72.73％ of the total number in the initial group, patients with the blood stasis syndrome occupied 75.61％ of the total number in the re-treatment group.③ The distribution of TCM syndromes was statistically different with the main symptoms (P＜ 0.05 ) . Conclusion All 516 cases of pulmonary tuberculosis had Yin deficiency. There were 380 patients had the stasis, occupying 73.64%.Therefore, the methods of nourishing yin and removing blood stasis should be used to treat patients with pulmonary tuberculosis.

Objective　To construct a recombinant plasmid pcDNA3-sHLA-G1 expressing soluble HLA-G1. Methods　 Total cell RNA was extracted from the cell line Jeg-3 and the cDNA was amplified by RT-PCR； The cDNA fragment was inserted into the eukaryotic expressing vector pcDNA3 and the recombinant plasmid was identified by restriction endonucleases digestion and sequencing. Results　 After restriction endonucleases treatment and sequencing, it was confirmed that the pcDNA3-sHLA-G1 had been constructed successfully. Conclusion　 In this study, the recombinant plasmid pcDNA3-sHLA-G1 had been constructed successfully.