Objective To explore the risk factors of female breast cancer and provide the basis for the etiology of female breast cancer,the identification of the high-risk population and the development of effective intervention measures.Methods The breast cancer patients who were confirmed by pathology were collected,and screened for healthy women whose age and living environment were similar to breast cancer patients.The questionnaire was used to collect risk-factors of breast cancer.The x2 test were used to analyze statistical difference of case group and control group.1 ∶ 2 logistic regression was used to analyze the relationship between risk factors and female breast cancer.Results There were 184 breast cancer patients and 368 healthy women in research.Benign breast disease,breastfeeding,abortion,depression,smoking,passive smoking,eating fried or smoked foods,drinking tea had statistical differences between case group and control group.And benign breast lesions(OR =1.553,95％ CI 2.045 ～ 10.924),abortion(OR =0.775,95 ％ CI 1.423 ～ 3.309),smoking(OR =0.674,95％ CI 1.009 ～ 3.817) passive smoking (OR =2.98,95％ CI 1.680 ～ 3.720) were the risk factors of breast cancer,and drink tea(OR =-0.425,95％ CI 0.446 ～ 0.958),eating soy products (OR =-0.687,95 ％ CI 0.284 ～ 0.889) breast-feeding (OR =-0.827,95％CI 0.193 ～ 0.991) were the protective factors of breast cancer.Conclusion The risk factors of female breast cancer in Binzhou are benign breast disease,abortion,passive smoking,depression,and the protective factors are drinking tea and breast-feeding.

Objective To study the correlation between the level of serum high sensitivity C-reactive protein (hs-CRP) and induced sputum cell counts,pulmonary function and asthma control test (ACT) in order to investigate the clinical application value of serum hs-CRP for the acute exacerbation bronchial asthma patients.Methods Thirty acute exacerbation bronchial asthma patients were collected as asthma group,14 healthy people during the same period were collected as control group.The fasting serum hs-CRP was examined in control group and before and after treatment of 5-7 d in asthma group.The induced sputum was obtained by ultrasonic atomizer 4.5％ sodium chloride,and the percentage of eosinophils (EOS％),neutrophil,macrophages,lymphocytes was calculated.The pulmonary function index was determined in control group and before and after treatment in asthma group.ACT score was performed in asthma group.The correlation between the serum hs-CRP and induced sputum differential leukocyte count,the pulmonary function index and ACT score was analyzed by Spearman correlation analysis.The effect of the serum hs-CRP and induced sputum EOS％ on inflammatory response of acute exacerbation bronchial asthma was evaluated by receiver operating characteristic (ROC) curve analysis.Results Two cases were excluded in asthma group.The serum hs-CRP before and after treatment in asthma group was higher than that in control group[4.18(2.12-11.12) and 3.40(2.02-8.91) mg/L vs.1.48(1.03-4.81) mg/L,H =18.939,P＜ 0.01].The serum hs-CRP before treatment was higher than that after treatment in asthma group (P ＜ 0.01).In asthma group,the serum hs-CRP had positive relationship with induced sputum EOS％ (r =0.849,P =0.000).There were negative correlations between the serum hs-CRP and the pulmonary function index such as one second forced expiratory volume percent predicted,one second forced expiratory volume occupied vital capacity ratio,maximum mid-expiratory flow percentage of predicted (r =-0.617,-0.559,-0.398,respectively,P =0.000,0.002,0.036,respectively).It was also found that the serum hs-CRP and ACT score had the remarkable Spearman linear correlation (r =-0.511,P =0.005).The area under the ROC curve of the serum hs-CRP was 0.713 (P =0.003).Conclusions There are significant correlations between the serum hs-CRP and induced sputum EOS％,pulmonary function,ACT score in the acute exacerbation bronchial asthma patients.It is found that hs-CRP could be considered as one of the potential marker to evaluate the systemic inflammation level of bronchial asthma.

Objective: To observe the clinical efficacy of pediatric Tuina (Chinese therapeutic massage) plus oral administration of Chinese medication for pediatric anorexia due to spleen failing in transportation. Methods: A total of 94 children with anorexia due to spleen failing in transportation were randomized into an observation group and a control group by the random number table method at a ratio of 1:1, with 47 cases in each group. The control group was treated with modified Bu Huan Jin Zheng Qi San, and the observation group was treated with additional Tuina manipulation of Yun Shui Ru Tu based on the treatment in the control group. After 4 weeks of treatment, the clinical efficacy of the two groups was observed, and the traditional Chinese medicine (TCM) symptom score, serum leptin, neuropeptide Y, and serum zinc and calcium levels were compared. Results: After 4 weeks of treatment, the total effective rate of the observation group was 87.2％, and that of the control group was 74.5％. The difference between the two groups was statistically significant (P<0.05). After treatment, the TCM symptom scores in both groups decreased significantly (P<0.05), and the scores in the observation group were lower than those in the control group (P<0.05). After treatment, the serum leptin level in both groups decreased, and the level in the observation group was lower than that in the control group (P<0.05). The serum neuropeptide Y, zinc, and calcium levels in both groups increased after treatment, and the levels in the observation group were higher than those in the control group (P<0.05). Conclusion: Tuina manipulation of Yun Shui Ru Tu plus oral administration of Chinese medication is significantly effective in treating pediatric anorexia due to spleen failing in transportation, which can improve the appetite of children and improve the symptoms of anorexia, and the curative effect is better than that of oral administration of Chinese medication alone.

AIM: To investigate the effects of intermittent hypoxic exposure and normoxic convalescence on the parameter of erythrocyte and serum hypoxia inducible factor1 alpha (HIF-1α) and erythropoietin (EPO)levels. METHODS: Rat models of intermittent hypoxic exposure were established, combined with the clinical research on volunteers experiencing the intermittent plateau work. Blood samples for red blood cell (RBC) counts, hemoglobin (Hb) and hematocrit (HCT) were collected, serum HIF-1α and EPO levels were measured using enzymelinked immunosorbentassy. RESULTS: RBC counts, Hb concentration and HCT were significantly higher than the normoxic group (P < 0.05), after exposure of rats to hypoxia from 7 to 28 days. Compared with the normoxic group, serum HIF-1α levels were higher in the group of IH3, 7, 14 days, and EPO had a corresponding increase in the group of IH3, 7 days. Then, a decrease was observed in parameter of erythrocyte and serum HIF-1α and EPO levels after 14 days normoxic convalescence treat. In volunteers studies, RBC counts in 8 monthes group and Hb concentration in 2 years group were significantly higher than the plain group (P < 0.05). The change of HCT was nearly the same as RBC, and HCT in 2 years group was higher than the plain group (P < 0.05). Compared with the plain group, EPO had no significant differences in any of plateau group. CONCLUSION: Intermittent hypoxic exposure can enhance serum hypoxia inducible factor-1 alpha and erythropoietin levels and the generation of red blood cells, which leads to an increase in hemoglobincon concentration and hematocrit. The results have changed with the hypoxic exposure period prolonged. Normoxic convalescence after intermittent hypoxic exposure can make the related indexes reduced, and contribute to the organism recovery.

Objective To investigate the genotypic characteristics of hepatitis B virus(HBV) in Tibet.Methods Serum samples from 60 cases of hepatitis B in Tibet Autonomous Region were collected from January 2000 to March 2004.HBV genotypes were analyzed by sequencing S region and precore/core region.Results HBV isolated from 60 cases were all found to be D genotype by S region sequencing.Dc mixture was found in 59 cases,showing the recombination between their precore/core gene and genotype B virus at 1804-2299 nucleotide.The other one case showed Dbc mixture genotype,showing recombination between its precore/core gene and genotype B and C genes.Conclusions All Tibet cases in this study show mixture genotype D with recombination with genotvpe C or both genotype B and C at precore/core region.No case of pure genotype D is found.

BACKGROUND:Growth-associated protein-43 is the primary substance for constructing plasticity of central nervous system, which is recognized as the first molecular marker for studying growth and repairing injury of nerves. OBJECTIVE: To explore the expression of growth-associated protein-43 in rats with focal cerebral ischemia following bone marrow mesenchymal stem cells (MSCs) transplantation.DEISNG, TIME AND SETTING: A randomized, controlled, animal study was performed at the Laboratory of Department of Neurology, Renmin Hospital of Wuhan University and Doctoral Scientific Research Work Station of C-BONS Pharmacy, Hubei, China, from March 2006 to October 2007.MATERIALS: Sixty adult Sprague Dawley rats, with clean grade, irrespective of genders, weighing (180±20) g, were randomly divided into the model, sham operated, and MSCs transplantation groups, with 20 animals in each group. METHODS: Additional 4 Sprague Dawiey rats, aged 2 months, were selected to isolate and culture MSCs, which was labeled with 5-Bromo-2,-deoxyuridine (BrdU). In the sham operated group, the right internal carotids were deligated after anaesthesia. The remained rats were prepared for models of right middle cerebral artery ischemia. At 24 hours after model preparation, 20 μL culture solution containing 5×105/L MSCs was injected into the left lateral ventricle of rats in the MSCs group, the same volume of phosphate buffer saline was injected in the model group.MAIN OUTCOME MEASURES: The rats were sacrificed prior to and at days 7, 14, and 21 after transplantation. The expression of growth associated protein-43 at the infarcted areas, the survival and migration of transplanted cells were examined by immunohistochemistry, at the same time, neurological deficit scores were recorded.RESULTS: The transplanted MSCs migrated from the left lateral ventricle to the infracted areas. Seven days after transplantation, the expression of BrdU-positive cells was found, reached a peak at day 14, and gradually decreased, until disappeared at 28 days after transplantation. Results of immunohistochemistry image analysis showed that immunological activity of growth associated protein-43 around the infarcted area of the MSCs group was obviously greater than that of the model group at days 7 and 14 after transplantation (P<0.06). There was no neurological deficit in the sham operated group. Moreover, with time prolonged, the neurological deficit scores gradually decreased in the model and MSCs groups, which were significantly lower in the MSCs group compared to the model group at day 14 after transplantation (P<0.05).CONCLUSION: MSCs transplantation up-regulates the expression of growth associated protein-43 around the infarcted area, which is consistent with the recovery of neurological function.

Objective To investigate the clinical significance of platelet activation markers CD62p and PAC-1 in patients with hypertension and coronary heart diease.Methods To select patients who were in the Second Affiliated Hospital of Dalian Medical University between January 2012 and September 2012,42 patients with hypertension,46 patients with coronary heart disease.Flow cytometry was used to test the positive percentage of the platelet surface activation markers CD62p and PAC-1 .To observe the difference among hypertension group,coronary heart disease group and health control group;hyper-tension and coronary heart disease group.Results The positive percentage of platelet activation markers CD62p and PAC-1 in hypertension (36.36±9.62,7.18±8.20)%,coronary heart disease group (42.74±14.60,8.81±12.53)% showed sta-tistically significant differences (t=4.150~5.853,P<0.01)compared with healthy control group (26.82±9.13 ,1.09± 1.05)%.The positive percentage of CD62p in coronary heart disease group (42.74±14.60)% was also higher than that of hypertension (36.36±9.62)%,and the difference between them was of statistical significance (t=2.444,P<0.05).Con-clusion The hypertensive patients and coronary heart disease were in pre-thrombotic state,the activation of platelet increase significantly.Molecular markers should be measured in hypertensive patients and coronary heart disease for prevention and treatment of thrombotic disease complication.

Objective To construct an RNAi lentiviral vector targeting rat CD86 gene and detect its effect of gene silencing in dendritic cells. Methods The effective sequence of siRNA targeting rat CD86 gene was confirmed in our previous work. DNA oligo containing short hairpin frame was synthesized and re-annealed, and then cloned into pGCL-GFP lentivind expression vector. PCR and sequencing analysis were made for verifying the positive clones. The virus packaging plasmids were transfected into 293T cells to har-vest shRNA lentivirus. After infection in dendritic cells, real-time PCR and Western blot were performed to determine the expression level of CD86 at mRNA and protein of NC( negative control virus). Results PCR and sequencing analysis revealed that shRNA plasmid was correctly constructed. Virus with a titer of 2×108 TU/ml was successfully packaged. CD86 expression in dendritic cells can be knockdown at both mRNA and protein level by virus infection as characterized by 90.6% decrease of mRNA and significant inhibition of protein compared with NC. Conclusion The recombinant lentiviral shRNA expressing vector targeting rat CD86 gene has been successfully constructed and packaged. CD86 mRNA and protein can be effectively down-regulated in dendritic cells.

Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

Objective To investigate the effect of implanted cortical electrical stimulation (CES) on the expression of Nissl bodies and growth-associated protein 43 (GAP-43) in the brain after ischemic injury,and its mechanism.Methods Models of middle cerebral artery occlusion (MCAO) were established in 23 male Sprague-Dawley rats.They were randomly divided into a CES group (CES,n=13) and a no stimulation group (NS,n=10) and electrical stimulators were implanted in both groups.CES was applied for 14 d in the CES group but not in the NS group.The expression of Nissl bodies and GAP-43 around the infarct were quantified using version 6.0 of the ImagePro Plus system.Results In the CES group the Nissl bodies had a deep color,and their percentage of area was higher than that in the NS group.The GAP-43 positive expression area also had a relatively deep color,and the average percentage of positive expression area was also higher than that in the NS group.Conclusions CES can enhance the expression of Nissl bodies and GAP-43 after cerebral infarction.This suggests that CES can promote axon growth and the formation of new neural circuits.