Objective Microheterogeneity is an important problem in molecularly biological and proteomic research on malignancies.The goal of this study was to investigate the method to purify targeted cells in lung cancer and its paired normal tissues for proteomic study on lung cancer applying laser capture microdissection(LCM).Methods LCM technique was emploged to obtain the cells of lung cancer tissues and their paired normal tissues from frozen sections.Results LCM can be applied to quickly and precisely obtain pure targeted cells subgroup or single cell without interstitium under the microscope.Conclusion LCM can tackle the problem of tissue heterogeneity in molecular analysis successfully.These results indicate that LCM has a potential as a tool in lung cancer proteomic research.

Objective To investigate the influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells.Methods The S100A6 gene RNA interference vector was transfected in A549 lung adenocarcinoma by lentivirus.The experiment was divided into three groups:pLenR-GPH group (the vector without S100A6 RNAi gene was transfected),negative control group (no vectors was transfected),and RNAi group (the vector with S100A6 RNAi gene was transfected).S100A6 mRNA and protein were detected using real-time PCR and Western blotting.The biological behavior including cell proliferation,invasion,cell cycle and cell apoptosis were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide,transwell,and flow cytometer,respectively.Results The expression of S100A6 mRNA of A549 lung adenocarcinoma cell line in RNAi group (0.009 ± 0.001) was significantly decreased than those in negative control group (0.049 ± 0.005) and pLenR-GPH group (0.030 ± 0.006),with statistically significant differences (t =57.56,P =0.000;t =48.21,P =0.000).The expression of S100A6 protein of A549 lung adenocarcinoma cell line in RNAi group (0.107 ± 0.002) was significantly decreased than those in negative control group (0.341 ± 0.005) and pLenR-GPH group (0.311 ± 0.006),with statistically significant differences (t =37.34,P =0.000;t =27.51,P =0.001).The ability of cell proliferation at 48 hours in RNAi group (0.230 ± 0.008) was significantly declined than those in negative control group (0.292 ± 0.038) and pLenR-GPH group (0.307 ± 0.013),with statistically significant differences (t =25.31,P =0.003;t =29.42,P =0.001).The number of transmembrane cells in RNAi group (11.40 ± 1.36) was significantly declined than those in negative control group (26.80 ± 1.83) and pLenR-GPH group (25.80 ± 1.93),with statistically significant differences (t =29.44,P =0.001;t =23.17,P =0.005).The cell proportion of S phase in RNAi group (28.26％ ± 0.38％) was significantly lower than those in pLenR-GPH group (44.73％±0.66％) and negative control group (45.15％ ± 1.69％),with statistically significant (t =63.69,P=0.000;t =71.55,P =0.000).Cell propotion of G2-M phase in RNAi group (26.99％ ± 0.29％) was signi-ficantly higher than those in negative control group (13.26％ ±0.49％) and pLenR-GPH group (12.41％ ± 0.46％),with statistically significant (t =56.31,P =0.000;t =51.39,P =0.000).The cell apoptosis proportion in RNAi group (8.90％ ±0.48％) was significantly higher than those in negative control group (5.84％ ±0.21％) and pLenR-GPH group (5.99％ ±0.37％),with statistically significant (t=51.34,P =0.000;t =47.27,P =0.000).Conclusion S100A6 gene involves the proliferation,invasion,cell cycle and apoptosis of tumor cells,which has close correlation with occurrence,development and metastasis of lung adenocarcinoma.S100A6 gene is hopeful to become a molecular target for the diagnosis and treatment of lung adenocarcinoma.

ObjectiveTo investigate the effects of Jiuqiang Naoliqing(JNQ) on the morphology of the vital organs(the brain,kidney and heart) in acute hypertension rats.MethodsThe model was established by L-NG-nitro arginine(L-NNA),which can inhibit the synthesis of nitric oxide(NO).Affected animals were administrated with high,medium or low dose of JNQ while nifedipine was used as the positive control.Effects on cardiac function and morphology of the vital organs were investigated.Results2 weeks after the establishment of the model,the affected rats had a 36% higher blood pressure compared with the control group.The acute hypertension rats showed significant pathologic changed in the brain,kidney and heart,while there was no obvious difference in the heart rate,electrocardiaogram,and blood pressure between the administrated groups.After treatment with JNQ,the contraction force and the oxygen extraction of the myocardiocytes were significantly reduced and the increasing velocity of the left ventricular pressure was decreased.The brain,kidney and heart showed improvement in pathology analysis to different levels.ConclusionAlthough it has no obvious effects on the heart rate and blood pressure in acute hypertension rats,JNQ can decrease the oxygen extraction in myocardiocytes and can obviously alleviate brains,kidneys and hearts pathologic changes.

Objective To explore the diagnostic value of radial ultrasound (EBUS) combined with virtual navigation (VBN) in peripheral pulmonary nodules.Methods Two hundreds and forty cases of peripheral pulmonary nodules(0.8 cm≤diameter≤3 cm) in the respiratory department of our hospital from July 2014 to July 2015 were included and according to the different guide de vices,which were divided into the radial ultrasound combined with virtual navigation group (EBUS+ VBN group),radial ultrasound group (EBUS group),virtual navigation group (VBN group) and control group.The diagnostic rates were compared among the four groups and among different sizes of lesion.The time of lesion location and operating time were also compared between the EBUS+ VBN group and EBUS group.Results Among 240 cases,the diagnostic rate in the EBUS+VBN group was highest(81.67％),and the diagnostic rates had statistically significant difference among the four groups (x2=19.344,P=0.00);the diagnostic rates of lesions less than 2 cm in diameter were lower than that of lesions ＞2 cm in the EBUS+-VBN group and EBUS group,but without statistically significant difference (x2 =2.04,3.40,P =0.15,0.07);the locating lesions time and operating times in the EBUS+ VBN group were shorter than those in the EBUS group,but the difference between them was not statistically significant (P=0.03,0.04).Conclusion EBUS combined with VBN could improve the diagnostic rate of peripheral pulmonary nodules and shorten the time of lesion location and operating time.

Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.

Objective To explore the learning and memory impairment and pathology in hippocampus in rats after spinal cord contu-sion. Methods 36 adult female Sprague-Dawley rats were randomly divided into sham group (n=18) and spinal cord injury group (n=18). Spinal cord injury model at T10 was established with modified Allen's technique (10 g × 25 mm). The hindlimbs behavior of rats was rated with Basso-Beattie-Bresnahan (BBB) scores once a week for 5 weeks. They were tested with motor evoked potentials (MEP) and Morris wa-ter maze 5 weeks after injury. The pathology of hippocampus was detected with HE staining 1 week, 3 weeks and 5 weeks after injury, 4 rats in a group, repectively. Results The BBB scores were significantly lower in the spinal cord injury group than in the sham group at each time point (P<0.05). The latencies of both N1 and P1 wave of MEP were significantly longer in the spinal cord injury group than in the sham group (P<0.001), while the amplitudes were significantly less (P<0.001). For the Morris water maze, the latency of arrival platform were sig-nificantly longer in the spinal cord injury group than in the sham group (P<0.001), and the time in target was significantly less (P<0.001), with more systematic positioning or annular positioning, while the sham group with more space-based positioning. Morphologically abnor-mal cells in hippocampus gradually increased since the first week after injury, with the decrease of cells survival, while it was normal in the sham group. Conclusion Spinal cord contusion can cause learning and memory impairment in rats, which may be related to injury in hippo-campus.

Robotics ; Robotic Surgical Procedures ; Kidney Transplantation