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 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.

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.

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.

This study was designed to evaluate the correlation between immune antibodies and pelvic inflammatory disease(PID)using retrospective analysis.Cases were selected from 171 patients who met the diagnosis of PID in Liuzhou People's Hospital of Guangxi Province from January 2022 to March 2023,and the PID patients were further divided into simple PID group(53 cases)and in PID combined with reproductive tract infection group(118 cases)according to the presence or absence of reproductive tract infections,while 83 cases of women who did not meet the specific diagnostic criteria of PID and did not have reproductive tract infections were selected as the control group during the same period.The positive rate of immune antibodies in the three groups were observed and compared to explore the relationship between immune antibodies and PID.Data showed that the positive rates of immune antibodies were significantly higher in the PID alone group and the PID combined with reproductive tract infection group than that in the control group.Furthermore,the positive rate of immune antibody TPOAb was significant difference in the PID combined with reproductive tract infection group and the PID alone group(P<0.05).In conclusion,TPOAb is closely associated with reproductive tract infections.

Objective:To investigate the facilitators and hindrances of smoking cessation in patients with chronic obstructive pulmonary disease (COPD), and to provide a basis for developing individualized smoking cessation intervention strategies for COPD patients.Methods:Based on the health ecology theory and using a phenomenological approach in qualitative research, purposive sampling was used to select 15 COPD patients with smoking history who were admitted to Tianjin Fourth Central Hospital from March to May 2023 and face-to-face semi-structured interviews were conducted. Colaizzi seven-step analysis was used to analyze the interview content.Results:A total of 15 COPD patients were interviewed, including 13 males and 2 females, aged 61-75 years old. The facilitators of smoking cessation in COPD patients included complications, sequelae of novel coronavirus infection, fear of death, smoking cessation counseling by medical staff, heavy family financial burden, and smoke-free environment. The hindrances of smoking cessation in COPD patients included milder disease symptoms of COPD, higher levels of nicotine dependence, false disease-related perceptions, family supervision and control, and occupational experience.Conclusions:Smoking cessation in COPD patients is influenced by five factors: personal characteristics, behavioral characteristics, interpersonal network, living and working conditions, and policy environment. Medical staff and relevant national institutions should formulate corresponding smoking cessation strategies according to address the facilitators and hindrances of smoking cessation in COPD patients, so as to further reduce the smoking prevalence of COPD patients, promote the health of patients and reduce the burden of disease.

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.