ObjectiveTo study the dose and time effects of X-ray radiation on the expression of Pokemon gene and protein in human lung adenocarcinoma cell line A549.MethodsA549 cells was exposed to different doses of X-ray (2,4,6 and 8 Gy),and the expression of Pokemon mRNA and protein of the cells was detected by using Quantitative real-time PCR and western-blotting at 2,4,8,12,24,and 48 h after irradiation. 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazoliumbromidewasused to detectthe proliferation of A549 cells at 1,2,3,4,and 5 d after 2 Gy X-ray irradiation.The mock treated A549 cells were used as the control.ResultsThe expression of Pokemon mRNA trended to decrease after irradiated with 4,6 and 8 Gy in the earlier period and increased in the later period with statistical difference at the most time points (t =3.40 -154.76,P =0.000 -0.041 ).The expression of Pokemon protein trended to increase and reached the peak at 8 h after irradiated of 2,4,6 and 8 Gy with statistical difference at the most time points ( t =4.18 - 89.64,P =0.000 - 0.039).Compared with the control,the proliferation of A549 cells was significantly inhibited during 3 to 5 d after irradiation of 2 Gy ( t =2.34 - 18.19,P =0.000 -0.040).ConclusionsX-ray irradiation may increase the expression of Pokemon mRNA and protein in A549 cells,which might be correlated with radiation-resistance of A549 cells.

Aim To set up a highly effective and automatic mouse sleep-wake bioassay system,and evaluate the system through analysis of the somnogenic effects of L-stepholidine(SPD),targeting at dopamine D1/D2 receptors in mice.Methods The animals were housed in an insulated and soundproof recording chamber maintained at a constant temperature and humidity on an automatically controlled 12 h light/12 h dark cycle.The electroencephalogram and electromyogram were recorded continuously for 48 hours and analyzed by SleepSign software.Saline was administered ip to the mice at 21:00 on the first day,and SPD was given on the next day at the same hour.The vigilance state was analyzed based on the polygraphic recordings by the same software.Results The system has been demonstrated to be highly efficient and stable in recording and reliable in analyzing sleep-wake behavior in mice.With the aid of the sleep bioassay system,we found that SPD significantly increased the total time spent in sleep during dark period,and prolonged durations of non-rapid eye movement sleep episodes,with a concomitant reduction in amount and EEG power density of wakefulness.SPD rendered no effect on rapid eye movement sleep.Conclusion Through the reliable mouse sleep bioassay system,we found that SPD promotes non-rapid eye movement sleep but not rapid eye movement sleep in mice.

Objective Compare the reproductive activity of pancreatic stem cells isolated from diabetic rat with that of normal rat to assess the effect of microenvirement on stem cells'proliferation.Methods Diabetic rat model were prepared by peritoneal injecting 60mg/kg streptozotocin.Pancreata from 10 diabetic rats and 10 normal rats were trimmed and digested into single cells which were then inoculated onto the culture plate respectively.After passaging 3 generation the cells were identified by nestin immunostaining.During culturing the cell growth curve was observed and the cell cycle of the third and seventh generation cultured cells was evaluated by flow cytometry.Results After passaging pancreatic stem cells was purified and identified by nestin immunostaining.The reproductive activity of diabetic rat was much better[ratio of s stage:(20.7?2.7)% vs(14.1?2.5)%,P0.05].Conclusions The stem cells are stimulated to proliferate after islet injure.The reproductive activity of cells can be provoked by the microenvironment.

Objective:To evaluate the effects of stellate ganglion block (SGB) on the activation of M1 microglia during cerebral ischemia-reperfusion (I/R) in rats.Methods:Fifty-four SPF healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 240-270 g, were divided into 3 groups ( n=18 each) using a random number table method: sham operation group (group Sham), cerebral I/R group (group IR) and SGB group.Blood vessels were only exposed, without occlusion in group Sham.Cerebral I/R was induced by occlusion of the middle cerebral artery for 90 min followed by reperfusion in group IR.Cervical sympathetic trunk transaction was performed to induce left SGB immediately after onset of reperfusion in group SGB.Blood samples were collected from the apex of the heart at 6, 12 and 24 h of reperfusion for determination of the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in the serum (using enzyme-linked immunosorbent assay). The animals were sacrificed after the neurological function was evaluated at 24 of reperfusion, and brain tissues were removed for microscopic examination of the pathological changes in cortex, for determination of percentage of cerebral infarct size (by TTC staining), for assessment of cell apoptosis and apoptosis rate in cortex (by TUNEL), and for determination of the expression of microglial biomarker Iba-1 and activated M1 microglia biomarker CD68 (by Western blot). Results:Compared with group Sham, the neurological function score, percentage cerebral infarct size, apoptosis rate in cortex, concentrations of TNF-α, IL-6 and IL-1β in the serum, and the expression of Iba-1 and CD68 were significantly increased in IR and SGB groups ( P<0.05). Compared with group IR, the neurological function score, percentage cerebral infarct size, apoptosis rate in cortex, concentrations of TNF-α, IL-6 and IL-1β in the serum, and the expression of Iba-1 and CD68 were significantly decreased in group SGB ( P<0.05), and the pathological changes of brain tissues were significantly attenuated in group SGB. Conclusion:The mechanism by which SGB reduces cerebral I/R injury is related to inhibiting activation of M1 microglia in rats.

The effects of epigenetic modification on the differentiation of islet cells and the expression of associated genes (Pdx-1, Pax4, MafA, and Nkx6.1, etc) were investigated. The promoter methylation status of islet differentiation-associated genes (Pdx-1, Pax4, MafA and Nkx6.1), Oct4 and MLH1 genes of mouse embryonic stem cells, NIH3T3 cells and NIT-1 cells were profiled by methylated DNA immunoprecipitation, real-time quantitative PCR (MeDIP-qPCR) techniques. The histone modification status of these genes promoter region in different cell types was also measured by using chromatin immunoprecipitation real-time quantitative PCR methods. The expression of these genes in these cells was detected by using real-time quantitative PCR. The relationship between the epigenetic modification (DNA methylation, H3 acetylation, H3K4m3 and H3K9m3) of these genes and their expression was analyzed. The results showed that: (1) the transcription-initiation-sites of Pdx-1, MafA and Nkx6.1 were highly methylated in NIH3T3 cells; (2) NIH3T3 cells showed a significantly higher level of DNA methylation modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIT-1 cells (P<0.05); (3) NIT-1 cells had a significantly higher level of H3K4m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIH3T3 cells (P<0.05), with significantly increased level of gene expression; (4) NIH3T3 cell had a significantly higher level of H3K9m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and with NIT-1 cell (P<0.05), with no detectable mRNA expression of these genes. It was concluded that histone modification (H3K4m3 and H3K9m3) and DNA methylation might have an intimate communication between each other in the differentiation process from embryonic stem cells into islet cells.

Objective In order to study the biological characteristics of macrophages and provide the materials to study the survival mechanism of intracellular parasites, we conducted this study to establish a high-purity alveolar macrophage isolation and culture method.Methods Goat lungs were lavaged with normal saline in sterile environment several times, and cells were collected and then goat alveolar macrophages were purified by density gradient centrifugation using peripheral blood mononuclear cells (PBMC) solution.The isolated goat alveolar macrophages were cultured in cell culture medium containing 10% fetal bovine serum and cell morphology was observed under an inverted microscope every day,and the phagocytic activity of the cells was detected by chicken red blood cell phagocytosis test.Flow cytometry was used to detect CD14, a characteristic monocyte-macrophage surface marker.Results The adherent cells were characterized by typical macrophage morphology, pseudopodia and protrusions, showing round and irregular shape, rich cytoplasm, and large cell body.Of the cultured macrophages, 54.5% could phagocytize chicken erythrocytes and showed good phagocytic activity.After one month of in vitro culture, 93.7% of the cells were able to express CD14 antigen, which had a macrophage-specific immunophenotype.Conclusions The alveolar macrophages obtained in this study have high purity and good bioactivity, thus provide a cell model for studying the immune mechanism of intracellular parasites.

Objective To explore the expression and localization of aquaporin 11(AQP11) in human term pregnancies with oligihydramnios, and its role in amniotic fluid balance.Methods We studied 55 patients who underwent elective cesarean sections, consisting of 25 patients with isolated oligohydramnios and 30 with normal amniotic fluid volume.Immunohistochemistry and reverse transcription polymerase chain reaction were employed to determine the expression and localization of AQP11 in the amnion, chorion and placenta.Results AQP11 protein was detected in expressed in the full-term pregnant women`s amnion, chorion and placenta.The expression in the amnion was positively correlated with amniotic fluid amount;the expression in the placenta was negatively correlated with amniotic fluid amount.The expression increased in the chorion with different amniotic fluid amount.Conclusion AQP11 plays an important role in regulating amniotic fluid balance.

Objective To analyze the medication rules of Professor ZENG Bin-fang’s treatment for chronic hepatitis B (CHB); To provide references for clinic. Methods Research data came from patients with CHB in outpatient and hospitalization in Department of Hepatology, Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region. In this study,data mining methods, such as complex network analysis and dot-mode mutual information, were utilized to conduct analysis on the syndromes, symptoms, treatment and medication of included patients, as well as the mutual relations. Results The study collected clinical data of 132 patients with CHB, who accepted 277 times of examination,and 277 prescriptions were included. High-frequency symptoms included:burnout and weakness, abdominal distension, hypochondriac pain, and lower limb edema; Common syndromes included:stagnation of liver qi and spleen deficiency, retention of damp-heat in the interior, liver and gallbladder damp-heat, liver qi stagnation, and qi-stagnation and blood stasis, etc., successively, including 126 times of examination and accounting for 45.5%of all cases. Common therapeutic methods involved in soothing liver and strengthening spleen, eliminating dampness and heat, nourishing liver and kidney, regulating vital energy and promoting blood flow and activating blood and activating spleen, successively. Dot-mode mutual information analysissyndrome-symptom relation:the differentiation point of stagnation of liver qi and spleen deficiency included burnout and weakness, thirst,dull pain of right lateral thorax, anorexia, and abdominal distension;syndrome-therapy relation:stagnation of liver qi and spleen deficiency contained the main therapeutic method of nourishing liver and strengthening spleen;syndrome-medicine relation: Bupleuri Radix, Codonopsis Radix, Scutellariae Radix, Angelicae Sinensis Radix, Paeoniae Radix Rubra,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Pinellinae Rhizoma Praeparata, raw Rehmanniae Radix, and Cyperi Rhizome. Analysis of complex network analysis showed that the core formula for stagnation of liver qi and spleen deficiency was Xiaochaihu Decoction and Siwu Decoction. Conclusion The treatment thinking of Professor ZENG Bin-fang for CHB pays attention to entire concept, attaches importance to Zang-fu transmission, regards“nourishing liver, soothing liver and strengthening spleen”as the therapeutic principle, together with nourishing yin, softening and resolving hard mass, clearing heat-toxin and eliminating dampness.

Objective To evaluate Zeus robot-assisted laparoscopic cholecystectomy. Methods Forty patients were divided into two groups receiving respectively Zeus robot-assisted laparoscopic cholecystectomy (group A, 20 cases), and laparoscopic cholecystectomy (group B, 20 cases). A variety of clinical parameters were evaluated. Compared between these two groups. Results Camera clearing (1.1?1.0) times and time used for operation field adjustment in group A (2.2?0.7) min were significantly less than those in group B (4.5?1.5,7.5?1.2) min. Dissection actions(337?86)times and operative errors(10%) in group A were less than those in group B(389?94) times,25%. The operation time(104.9?20.5) min and setup time (29.5?9.8) min in group A were longer than those in group B (78.6?17.1) min,(12.6?2.5) min. The blood loss,and postoperative hospital stay were similar. There were no postoperative complications in either groups, and conversion to open surgery was done in one each patients. Conclusions Compared with laparoscopic technique, Zeus robotic surgical system offers greater ability of controlling operation field, precise and stable operative manipulations though it requires longer operation time.

Objective To explore the application value of 3.0T MR diffusion tensor imaging (DTI)in 3D reconstruction of the knee peripheral nerve.Methods DTI with 25 motion probing gradients (MPGs)was performed on the left knee in 22 healthy young volunteers;By choosing the original images scaned in the vertical direction,three post-processing methods including maximum intensity proj ection (MIP),volume rendering (VR)and diffusion tensor tractography (DTT)were used.Two radiologists evaluated these images visually.For the same nerve,MIP,VR and DTT were compared using Friedman test.Intraclass correlation coefficient (ICC )analysis was used to compare the correlation among the three imaging methods.Score consistency between two observers was calculated by using the Kappa statistic.Results For the same nerve,there was no significant statistical difference for the MIP,VR and DTT imaging quality scores(P>0.05).The I CC value and the k value were 0.718-0.856,0.668-0.705,respectively.Conclusion Stereoscopic and clear displaying the tibial nerve/the common peroneal nerve in the knee can be achieved by using MIP,VR and DTT post-processing methods based on DTI original images.The reconstruction of MIP,VR is simple and efficient,in which the objectivity is better than DTT.The reconstruction of MIP,VR can be used as effective preview methods before the DTT reconstruction with complexity and strong subjective dependence.