SYBR Green I Real Time-qPCR method was developed to quantify the numbers of copyies of AlkB ( alkanes degradation gene) and Nah ( naphthalene dioxygenase degradation gene) functional degradation gene corresponding to alkanes and aromatic hydrocarbons degradation. Two pairs of primers AlkBf/AlkBr and Nahf/Nahr were designed for AlkB and Nah amplification respectively, according to the nucleotide sequences of related degradation microorganisms published in GenBank. The purified recovery products of traditional PCR were combined with pEASY-T1 vectors and transformed in competent cells to amplify. The recombinant plasmids were extracted and used as positive templates to create standard curve through gradient dilution. The conditions for the real time PCR were as the follows: the final concentration of forward and reverse primers were 0. 2 μmol/L, 2×TransStart Top Green qPCR SuperMix, and the annealing temperatures of AlkB and Nah PCR were 50℃ and 57℃, respectively. The method showed a sensitivity of 100 times higher than that of the traditional PCR method and good repeatability. The numbers of copies of AlkB in three functional regions of an oilfield indicated that oil producing zone with serious oil pollution had the highest AlkB copy numbers, and residential zone with lighter oil pollution had the lowest AlkB copy numbers. Nah degradation gene distribution was more uniform.

Chronic intermittent hypoxia ( CIH ) caused by ob-structive sleep apnea hypopnea syndrome( OSAHS) is an impor-tant factor causing or aggravating many kinds of cardiovascular and cerebrovascular diseases. Establishing a rational animal model for intermittent hypoxia is an essential method to study the CIH related cardiovascular diseases. Recently, researchers have tended to simulate intermittent hypoxia condition by controlling the oxygen concentration of the environmental air around the ani-mals. In the paper, we summarize and compare the methods of making intermittent hypoxia animal model in recent literature, from aspects of experimental animals, gas control apparatus, gas species and concentration, intermittent hypoxia treatment time, and anoxic cycle mode.

Objective To study the change characteristics of erythrocyte parameters in carriers of deletional alpha-thalassemia gene.Methods 389 patients with deletional alpha-thalassemia gene determined by the gap-PCR technique were classified into three groups based on different genotypes of alpha-thalassemia including silent thalassemia group,alpha-thalassemia trait group (265 ca-ses)and intermediate thalassemia group,and contemporaneous 188 healthy adults were randomly selected as the normal control-group(NC).The erythrocyte parameters including RBC,Hb,MCV,MCH,RDW were retrospectively analyzed and their differences were compared among aboved-mentioned groups by the analysis of variance and the multiple comparison.Results Alpha-thalasse-mia manifested by different degrees of microcyte hypochromia.There were statistically significant differences in the erythrocyte pa-rameters among various genotypes and phenotypes about erythrocyte indices(P <0.05).Moreover,the Hb,MCV and MCH values were lower than those in the control group,and had the decreasing tendency with the increase of deletedα-globin gene numbers;but the RDW value was higher than that in the control group,showing the increasing tendency,the differences had statistical siginifi-cance(P <0.05).Conclusion With the increase of deleted α-globin gene number,the characteristic of small-cell low-hemoglobin is more and more remarkable.RBC has a overall increase,while Hb,MCV and MCH have a decreasing tendency,and the heterogeneity of erythrocyte volume is increased.When MCV and MCH decreasing in high risk region,thalassemia should be highly suspected, but normal MCV and MCH can not exclude silent thalassemia and alpha-thalassemia trait.

Objective To evaluate the relationship between the concentration of pentane in the exhaled air and degree of the lung injury in non-heart-beating (NHB) rabbits.Methods Twenty-four healthy male Japanese white rabbits weighing 2.4-3.0 kg were randomly divided into 4 groups ( n =6 each):A,B,C and D groups.The NHB model was established by exsanguination through the femoral artery.The exhaled gases were collected and lung tissues were removed at 0,30,60 and 120 min after cardiac arrest in A,B,C and D groups respectively.The concentration of pentane in the exhaled gases was detected immediately using the gas chromatography-mass spectrography.The wet to dry (W/D) lung weight ratio and content of malondialdehyde (MDA) in lung tissues were measured.The lung injury score (LIS) was recorded.The maximal volume ( Vmax ) of the lung was recorded when the airway pressure reached 30 cm H2O.Results Compared with groups A and B,the exhaled pentane concentration was significantly increased in group C,and the W/D ratio,content of MDA and LIS were significantly increased,while Vmax was significantly decreased in group D ( P ＜ 0.05).Compared with group C,W/D ratio and LIS were significantly increased in group D ( P ＜ 0.05 ).Conclusion The concentration of exhaled pentane can not reflect the degree of the lung injury in NHB rabbits.

The clinical data of 143 patients with type Ⅱ a hepatolithiasis with normal function of the sphincter of Oddi were analyzed.The patients were divided into two groups at random.The data of the 2 groups of patients were compared on the operating time,hospitalization,rates of complication,residual stone rates and recurrence rates.The A group patients underwent laparoscopic video choledochoscopic hepaticocholangiolithotomy,T-tube drainage and choledochoscopic lithotomy.The B group patients underwent laparoscopic video hepatectomy and choledochoscopic lithotomy.For most of the patients in the A group,the treatment had the advantages of less trauma,less complications and quicker recovery.There was no significant differences in residual stone rates and recurrence rates between the two groups.As it was difficult to remove stones in the caudate lobes of Ⅱ a type patients in A group due to the sharp angle between the hepatic duct and bile ducts of the caudate lobe,individualized programs should be adopted according to the location of stones on individual patients.

Objectives:To investigate the predictive effect of regional homogeneity (ReHo) from resting-state functional magnetic resonance imaging (rs-fMRI) in acute phase on memory function of chronic phage in ischemic stroke patients and the effects of residual learning (REL) on the predictive performance of machine learning models.Methods:From June 2019 to June 2021, rs-fMRI data of one-week after stroke (acute phase) were collected from 35 first-time ischemic stroke patients, and their memory scores were assessed by the Rey auditory verbal learning test (RAVLT) at 6 months after stroke (chronic phase). Using ReHo from rs-fMRI data in acute phase of ischemic stroke patients, the support vector regression (SVR) and the REL-based SVR (REL-SVR) were constructed to predict the patients’ memory scores at 6 months after stroke, and the performance of the two models was compared using Pearson correlation coefficient.Results:Based on the ReHo from acute phase, the correlation coefficient between the predicted values and the true scores from the SVR model was r=0.524, P=0.001, while the correlation coefficient obtained by the REL-SVR model was r=0.671, P<0.001. Brain regions with relatively higher weights such as Temporal_Pole_Mid_R (weight value: 1.03), Temporal_Mid_R(weight value: 1.03), Temporal_Inf_R (weight value: 1.03), Occipital_Mid_R (weight value: 0.57), Frontal_Mid_L (weight value: 0.32), Frontal_Sup_Medial_L (weight value: 0.53), SupraMarginal_L (weight value: 1.54), Calcarine_L (weight value: 0.65), Lingual_L (weight value: 0.58), Cuneus_L (weight value: 0.65), Precuneus_L (weight value: 0.83), cerebellum(weight value>1.0) made larger contributions to the prediction model. Conclusions:ReHo in the acute-phase can effectively predict memory in the chronic phase of ischemic stroke patients. Furthermore, REL can improve the performance of the traditional SVR model and achieve higher predictive accuracy.

Objective: To examine college students awareness of AIDS (acquired immunodeficiency syndrome) and HIV (human immunodeficiency virus), as well as their willingness to undergo testing, and to provide guidance for further education targeted towards AIDS prevention. Methods: The respondents were selected from two companies of military training camps in 4 universities in Fengtai District of Beijing using cluster sampling, and a questionnaire was used to obtain relevant information among 1 248 college freshmen. The content of the questionnaire included basic information about the students, awareness of AIDS, and willingness to undergo testing. Results: A total of 87.18% students were familiar with AIDS related knowledge, and 62.98% students intended to have HIV tests in the future. Willingness to be tested for HIV was higher among not local students (67.39%) than among local students(55.65%)(χ 2=17.32, P<0.05). The willingness to get HIV testing was higher among students who had an understanding of AIDS (65.26%) than among those who lacked an awareness(47.50%)(χ 2=18.87, P<0.05). In terms of the willingness to be tested for HIV, the main concerns focused on personal privacy (23.24%) and the cost (18.59%). Multivariate regression analysis showed that improving students awareness of five of the items related to a basic knowledge of AIDS may increase their willingness to get HIV testing(P<0.05). Most students indicated a preference to get HIV testing at a hospital (68.51%) or at the Centers for Disease Control and Prevention(42.79%). Conclusion The willingness to get HIV testing can be increased by launching an AIDS health education program that targets weak knowledge points with respect to AIDS awareness.

Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.

Objective:To investigate the changes in gray matter volume (GMV) and abnormalities in structural covariant network (SCN) patterns in patients with chronic pontine infarction (PI).Methods:Patients with unilateral chronic PI (case group) with the first onset admitted to the First Affiliated Hospital of Zhengzhou University and Tianjin Medical University General Hospital from October 2014 to June 2021 were prospectively included. At the same time, healthy subjects matched with age, gender and education years (normal control group) were included. High-resolution three-dimensional T 1 structural MRI images and behavioral scores of the subjects were collected. The voxel-based morphometry and two-sample t test were used to explore the differences in GMV between the groups. Using GMV differential brain regions as seed points, SCN was constructed to explore the abnormality of structural covariant patterns in patients with PI. Spearman rank correlation analysis was used to analyze the correlation between GMV in differential brain regions and behavioral scores. Results:A total of 60 patients with PI were enrolled, including 33 left PI and 27 right PI, while 34 healthy controls were also enrolled. Compared with the normal control group, the GMV in bilateral posterior cerebellar lobe decreased significantly in the left PI group, and the GMV in left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe decreased significantly in the right PI group (Gaussian random field correction with voxel level P<0.001 and cluster level P<0.05, cluster voxel >20), and there was a significant correlation between GMV values in the left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe and the motor function score ( P<0.05). In addition, compared with the normal control group, the right PI group had broader covariate brain regions and a significant increase in the number of structural connections between covariate brain regions (family-wise error correction with voxel level P<0.05, cluster voxel >20). Conclusions:The GMV in bilateral posterior cerebellar lobe decreases significantly in patients with chronic PI, and were secondary to broader covariate brain regions and structural connections. This may be the neural mechanism of impaired behavioral function in patients with PI.