Objective In order to research the effect of tyrosine kinase inhibitor in combination with radiotherapy on the inhibi‐tion of breast cancer cells and breast cancer bearing nude mice .Methods Methyl thiazolyl tetrazolium (MTT) assay was used to e‐valuate the inhibition of different concentrations of TKI on breast cancer cells ,the breast cancer cells was divided into three groups :the TKI treatment group ,the cells in the control group (no the TKI processing) and the control group (non‐TKI and X‐ray irradia‐tion group) .The sensitivity of the cells in each group to X‐ray was compared by colony formation assay .MCF7 cells were xenograf‐ted in athymic nude mice to establish the animal model ,which was used to evaluate the effect of anti‐cancer .Results Colony form‐ing test showed that the separated use of any concentrations of the TKI inhibitor could inhibit the breast cells ,and the cell viability was significantly reduced;TKI combined with X‐ray irradiation could significantly increase the sensitivity of breast cancer cells to radiation ,and the difference was statistically significant(P<0 .05) .Compared to TKI inhibitor or X‐ray irradiation alone ,the combi‐nation of TKI inhibitor with X‐ray irradiation could inhibit the growth of tumor effectively .Conclusion The TKI inhibitor in com‐bination with radiotherapy can effectively inhibit the growth of breast cancer cells ,which provides a new theoretical basis for the im‐plementation of the clinical breast cancer radio sensitization .

Objective To evaluate the association of COX2 genetic variants with the risk of esophageal cancer and the interaction of COX2 genetic variants with Hp infection. Methods A total of 119 patients with esophageal cancer and 238 frequency-matched controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism method. Odds ratios (OR) and 95% confidence intervals ( CI) were estimated by logistic regression. Results Case-control analysis showed an increased risk of developing esophageal cancer for 1195 GA(OR =2.69,95% CI= 1. 46-5. 14) and 1195AA ( OR = 2. 30,95% CI = 1.23-4. 89) genotype carriers,respectively, compared with non 1195 GG carriers. When stratified by Hp status, the significantly increased risk of esophageal cancer was found among Hp carrier with OR (95%CI) =2.74 (1.35-5.96) ,but not among Hp non-carriers. Conclusion Genetic polymorphism in COX2 promoter region may play an important role in esophageal cancer by Hp infection.

Objective To continuously draw blood from caudal vein of SD rats which controlled by the homemade holder with an im-proved method, and the effects of this method was evaluated according to the relevant indicators. Methods The 24 SD rats should be drew the blood from the caudal vein by the improved disposable infusion needle and supplied physiological saline at the condition of ether inhala-tion anesthesia and once a day until to the thirteenth day,and then the times of draw blood and the number of survival rats only to calculate the success rate and survival rate of rats were record. Results In the 12 days,the tail vein blood of 24 SD rats were successfully collected. But a rat died of excessive ether inhalation anesthesia on the thirteenth day. The success rate was 99. 68% and the rat survival rate was 95. 83%. Conclusion The method that applying an improved disposable infusion needle and supplying physiological saline is safe and ef-fective,flexible and convenient,which is suitable for experimental study that needs to draw blood from caudal vein of the SD rat at the contin-uous time.

Objective To establish a rat model of nonalcoholic steatohepatitis (NASH),and to investigate the preventative and therapeutic effects of compound ginkgo extract against NASH.Methods A total of 60 male Sprague-Dawley rats were fed with high-fat feed and 10％ fructose water for 24 weeks to establish the rat model of NASH.The general behaviors of the rats were observed,and the body weight was recorded.Blood samples from the inferior vena cava and the liver were collected after the last administration to measure serum total cholesterol (TC),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C),and low-density lipoprotein cholesterol (LDL-C),as well as liver function parameters.The liver index was calculated,HE staining was performed to observe liver histopathological changes,and the total lipase activity and the levels of TC,TG,and free fatty acid (FFA) in liver tissue were measured.Results After 24 weeks,compared with the normal group,the model group had a significantly faster increase in body weight,significant increases in serum levels of TC (2.20±0.52 mmol/L),TG (0.87±0.22 mmol/L),LDL-C (1.22±0.50 mmol/L),alanine aminotransferase (ALT) (129.4±44.7 U/L),and aspartate aminotransferase (AST) (209.3±42.8 U/L),liver index (3.62％±0.28％),and the levels of TC (4.42±1.39 mmol/mg.prot),TG (0.85±0.11 mmol/mg.prot),and FFA (644.78±36.65 μmol/L) in liver tissue,and significant reductions in serum HDL-C level (0.58±0.11 mmol/L) and the activity of lipoprotein lipase (LPL) (9.95±1.64 U/mg.prot)and hepatic lipase (HL) (9.91±1.03 U/mg.prot) (all P ＜ 0.01).In addition,the pathological results showed severe hepatocyte steatosis,varying degrees of inflammatory cell infiltration,exudation in the portal area,and necrosis of liver cells in the model group.After the intervention with compound ginkgo extract,there were significant reductions in serum levels ofTC (1.78±0.21 mmol/L),TG (0.58±0.07 mmol/L),LDL-C (0.84±0.19mmol/L),and ALT (84.1±17.1 U/L),AST (155.4±20.9 U/L),liver index (2.71 ％±0.15％),and the levels of TC (2.24±1.02 mmol/mg.prot),TG (0.46±0.11 mmol/mg.prot),and FFA (580.56±50.63 μmol/L) in liver tissue,as well as significant increases in serum HDL-C level (0.68±0.10 mmol/L) and the activities of LPL (15.54±2.21U/mg.prot) and HL (11.92± 1.87 U/mg.prot) (P ＜ 0.05 or P ＜ 0.01).At the same time,it significantly reduced hepatomegaly in rats and improved fatty degeneration and degree of inflammation in liver cells.Conclusion Compound ginkgo extract can prevent and treat NASH by correcting dyslipidemia,improving liver function and fatty degeneration in hepatocytes,and reducing the degree of inflammation,and its mechanism of action may be associated with increasing total lipase activity,reducing FFA in the liver,increasing the decomposition of TG,and reducing the synthesis of TG.

OBJECTIVETo investigate the association of TNF-β 252A >G variant with the risk of non-small cell lung cancer (NSCLC).

METHODSTotal 956 patients with NSCLC were recruited between January 2000 and December 2008 at Cancer Hospital, Chinese Academy of Medical Science as the case group, and 994 frequency-matched controls were randomly selected from a pool of cancer-free subjects recruited from a nutritional group. All the participants were unrelated Han Chinese. There were no age, gender restrictions. Smoking status of the subjects was surveyed. Informed consent was obtained and 3 ml peripheral blood was collected from each subject. All samples were genotyped by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). The OR and 95%CI were estimated by logistic regression to evaluate the relationship between TNF-β 252 A/G variant and the risk of lung cancer.

RESULTSThe frequencies of TNF-β 252 AA, AG and GG genotype were 30.9% (307/994) , 47.4% (471/994) and 21.7% (216/994) in lung cancer cases and 35.7% (341/956) , 48.1% (460/956) and 16.2% (155/956) in controls. Logistic regression analysis revealed that TNF-β 252 GG genotype contributed to a decreased risk of developing NSCLC (OR = 0.64, 95%CI: 0.49-0.83) compared with AA genotype. When stratified by smoking status, the individuals with 252 GG genotype had a significant increased risk of NSCLC (OR = 1.54, 95%CI:1.00-2.39) among smokers; which was less than those with AA genotype among smokers (OR = 2.88, 95%CI:1.91-2.24). When further stratified by smoking index, individuals with 252 GG genotype had a significant decreased risk of NSCLC among heavy smokers with OR (95%CI) of 2.24 (1.33-3.74), which was less than those with AA genotype (OR = 4.62, 95%CI:2.88-7.41).

CONCLUSIONTNF-β genetic variant may interact with environment factor to contribute to the susceptibility to NSCLC.

Asian Continental Ancestry Group ; Carcinoma, Non-Small-Cell Lung ; Case-Control Studies ; China ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lung Neoplasms ; Lymphotoxin-alpha ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Risk ; Risk Factors ; Smoking

Objective: To investigate the clinical features and pathogenic genes of a family with osteosclerosis. Methods: Six patients and six family members from a family in Jiangsu were tested for biochemical parameters, bone metabolic markers, bone mineral density, thoracolumbar anterior lateral slices, skull positive lateral radiographs, and pelvic plain films. Meanwhile, Sanger sequencing was performed to detect gene mutations of the proband and five other family members with high bone mass. The conformation of the mutational low-density lipoprotein receptor-related protein 5 (LRP5) protein was predicted by SWISS-MODEL. Results: Four adult patients (one male and three females) were tall, with mandibular enlargement and kyphosis in the center of the lower jaw, and none of the four had fractures. Their X ray examination revealed that the skull and long bone cortex was thickened, while the sella and mandible was enlarged. In addition, the absolute values of bone mineral density at each site of all patients were significantly higher as compared with the standard age- and sex-matched adults or adolescent mean reference values, with Z scores of L2-4, femoral neck and total hip being (6.31±4.03) SD, (6.56±2.36) SD, and (7.19±2.03) SD, respectively. The results of genetic sequencing revealed that all six patients carried a heterozygous mutation (c.331G>T; D111Y) in exon 2 of LRP5 gene, while other family members showed wild type (c.331G>G; D111D). Functional prediction indicated that this mutation was located at the amino acid terminal of exon 2 of LRP5 gene, which encodes the first β-helix-generating region of LRP5 protein. Conclusion The D111Y mutation in LRP5 gene leads to a clinical phenotype characterized by benign increased bone mineral density without increasing the risk of fracture. This mutation may further affect the downstream Wnt signaling pathway by altering the spatial structure of LRP5 protein, thereby promoting maturation and differentiation of osteoblasts and resulting in osteosclerosis.

ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine. MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats （SHR）. In this experiment， 50 SHR rats were randomly grouped as follows： model （n=8）， captopril （0.01 g·kg^-1） （n=6）， low-dose blank Fufang Lingjiao Jiangya pills （0.342 g·kg^-1） （n=6）， high-dose blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1） （n=6）， low-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， high-dose antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）， low-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.378 g·kg^-1） （n=6）， and high-dose goat horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1） （n=6）. Additionally， 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration， 3 h post administration， and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions， 48 SHR rats were randomly grouped as follows： model， blank Fufang Lingjiao Jiangya pills （0.684 g·kg^-1）， antelope horn-containing Fufang Lingjiao Jiangya pills （0.756 g·kg^-1）， 2× goat horn-containing Fufang Lingjiao Jiangya pills （0.824 g·kg^-1）， 4× goat horn Fufang Lingjiao Jiangya pills （0.969 g·kg^-1）， and 6× goat horn Fufang Lingjiao Jiangya pills （1.112 g·kg^-1）. The normal group included 8 WKY rats， and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks， and blood pressure was recorded at various time points （pre-administration， 3 h post administration， and on days 4， 7， 10， and 14 of administration）. Serum levels of angiotensin-converting enzyme （ACE）， angiotensin Ⅱ（Ang Ⅱ）， renin， and interleukin-6 （IL-6） were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart， kidney， and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2， angiotensin Ⅱ type 1 receptor （AT1R）， and angiotensinogen （AGT） in the kidney tissue were determined by Western blot， while the expression of nuclear factor （NF）-κB p65 and Toll-like receptor 4 （TLR4） in the thoracic aorta tissue was assessed by immunohistochemistry. ResultsCompared with the model group， all treatment groups showed lowered blood pressure （P<0.05， P<0.01）， and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group， the model group showed elevated serum levels of ACE， Ang Ⅱ， renin， and IL-6， while the elevations were declined in the Fufang Lingjiao Jiangya pills groups （P<0.05， P<0.01）. Pathological changes in the heart， kidney， and thoracic aorta were alleviated in all the treatment groups， with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R， AGT， NF-κB p65， and TLR4 and up-regulated protein levels of ACE2 （P<0.05， P<0.01） compared with model group， with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect. ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system （RAAS） and TLR4/NF-κB signaling pathways.