Objective:To clone and characterize the 16S rRNA of six species in the bacteria infecting respiratory tract to make gene chip.Methods:The primers of the target gene were designed and synthesized,and then the aimed fragment of the 16s rRNA was amplified by PCR and cloned.Finally the recombinant plasmids were characterized.Results:(1)The 16s rRNA gene of six species of bacteria was amplified.It was found that the size of amplified product by PCR was 1 300 bp in E.coli,S.aureus,S.pneumoniae,K.pneumoniae and H.influenzae and that of 1 100 bp in P.aeruginosa.(2)The JM109 transferred by the recombinant plasmid pMD18-T grew in Ampr culture was white colonies.(3)The specific bands could be found by restriction endonuclease and PCR analysis. (4)The sequence of the six bacterial 16s rRNA showed the same as those in the GenBank.Conclusion:The 16s rRNA of six species of bacteria is successfully amplified and cloned into plasmid pMD18-T. It will provide the basis for making gene chip detecting the six species of bacteria infecting respiratory tract.

Objective To investigate the effect of Xuebijing injection on inflammatory factors in patients with Stanford B aortic dissection (AD) after endovascular repair and approach its clinical significance.Methods Sixty patients with Stanford type B AD for endovascular repair admitted to the Department of Cardiothoracic Surgery of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine (TCM) from January 2014 to June 2016 were enrolled, and they were divided into a Xuebijing group and a control group according to the random number table method, 30 cases in each group. The patients of Xuebijing group received 100 mL Xuebijing injection+ 50 mL normal saline intravenous drip during operation, while the patients of control group received an equal volume of normal saline, 2 times a day in both groups for consecutive 3 days. Peripheral venous blood was collected before and after treatment for 1, 2, and 3 days, and the levels of C-reactive protein (CRP), tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA), and the prognosis of the two groups was observed.Results With the prolongation of time, the postoperative levels of CRP, TNF-α and IL-6 in the two groups showed a tendency of first increase and then decrease compared with the preoperative levels, the above indexes of the two groups were all significantly higher, CRP (mg/L) and TNF-α (ng/L) peaked on the 2nd day postoperatively (control group: CRP was 146.34±13.92 and TNF-α was 72.22±7.63, Xuebijing group: CRP was 114.92±9.89 and TNF-α was 53.44±6.80, allP < 0.05), however, IL-6 (ng/L) peaked on the 1st day postoperatively (control group: 146.08±10.29, Xuebijing group: 117.88±8.84), then decreased, all reached the valley on the 3rd day postoperatively (control group: CRP, TNF-α, IL-6 was 112.59±8.54, 43.73±4.10, 70.03±4.66 respectively, Xuebijing group CRP, TNF-α, IL-6 was 87.75±7.67, 39.43±4.63, 56.65±3.27, respectively), and at the same time point, the level of CRP, TNF-α, IL-6 in the Xuebijing group were significant lower than those in the control group, the change were the most significant on 3 days after operation [CRP (mg/L): 87.75±7.67 vs. 112.59±8.54, TNF-α (ng/L): 39.43±4.63 vs. 43.73±4.10, IL-6 (ng/L): 56.65±3.27 vs. 70.03±4.66, P < 0.05]. There were no serious complications such as renal failure, severe hypoxemia, infection, paraplegia, stent shift and so on, no hospital death occurred and all patients were discharged in rehabilitative condition. Conclusions Endovascular repair in patients with Stanford type BAD may activate an inflammatory response inducing the release of a large amount of inflammatory factors during the early postoperative period, Xuebijing injection can inhibit the inflammatory reaction and prevent the occurrence of postoperative complications.

Objective To study the CT performances and literature review of myoepithelioma (ME)of the parotid gland,to improve the recognition and diagnostic accuracy of this disease.Methods The CT findings of 1 2 cases with ME of the parotid gland confirmed by surgery and pathology were analyzed retrospectively,the characteristics of the age of onset,gender,size,location,density,boundary and enhancement were analyzed.And then,the related literatures were reviewed.Results Among the 1 2 patients,5 were male and 7 were female,age ranged from 11 to 68 years old,with a median age of 45.2 years.2 patients had the symptom of tenderness and fever, and the other 10 patients were asymptomatic and painless preauricular mass,2 of which were gradually enlarged.All the tumors were single in this study,2 had visible capsule infiltration or unclear boundaries,and other 10 had intact or smooth edges.CT showed that the masses all located in the parotid gland(1 in the deep lobe and 11 in the superficial lobe).The diameter of the tumor was 10 mm to 50 mm,with an average of 29.1 mm.The density was uniform in 2 cases and irregular cystic degeneration and necrosis in the other 10 cases.Mild enhancement was found in the arteriovenous phase after enhanced scanning,in which 8 cases had marginal enhancement or nodular enhancement in the arterial phase,and gradual filling into the center in the venous phase.Conclusion The CT performances of ME of the parotid gland have define characteristics.The possibility of myoepithelioma of the parotid gland should be taken into account when it has continuous centripetal reinforcement after enhancement,combing with the single cystic solid mass with clear boundary in superficial lobe of the parotid gland.Malignant myoepithelioma (MM)should be considered when the tumor envelope is incomplete or the boundary with the surrounding structure is not clear.

Objective To investigate the preventive effect of Xuebijing injection on acute lung injury induced by cardiopulmonary bypass (CPB) and the underlying mechanism. Methods ① In vivo experiment: 30 Sprague-Dawley (SD) rats were randomly divided into sham group, CPB group, Xuebijing pretreatment group (XBJ+CPB group) with 10 rats in each group. CPB model was reproduced in rats; and CPB was not performed in sham group, but only through arteriovenous puncture. In the XBJ+CPB group, 4 mL/kg Xuebijing injection was injected intraperitoneally 2 hours before CPB, sham group and CPB group were injected with equal volume of normal saline at the same time. The blood from femoral artery was analyzed 4 hours after operation, and the oxygenation index (PaO2/FiO2) was calculated. Then the rats were sacrificed to collect bronchoalveolar lavage fluid (BALF), and the lung permeability index (PPI) was calculated. The lung tissues were harvested, and the wet/dry weight ratio (W/D) of lung tissue was measured. The index of quantitative evaluation of alveolar injury (IQA) was measured. The levels of interleukins (IL-1, IL-6) and tumor necrosis factor-α(TNF-α) in lung tissue and BALF were measured by enzyme-linked immunosorbent assay (ELISA). The content of malondialdehyde (MDA) and the activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in lung tissue were detected by biochemical method. The microRNA-17-5p (miR-17-5p) expression in lung tissue was determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR).② In vitro experiments: type Ⅱ alveolar epithelial cells (AECⅡ) were cultured in vitro, and they were randomly divided into control group (the cells were treated by preoperative serum of CPB in patients with ventricular septal defect), CPB group (the cells were treated by serum after CPB in patients), and XBJ+CPB group (Xuebijing injection 10 g/L+serum after CPB in patients). After 12 hours of culture in each group, the expression of miR-17-5p was detected by RT-qPCR. AECⅡ cells were transfected with miR-17-5p mimic, inhibitor or corresponding control oligonucleotide (negative control), respectively, to observe the effect of miR-17-5p on Xuebijing regulating CPB-induced apoptosis rate and caspase-3 activity. Results ① In vivo experiment: compared with the sham group, the PPI, lung W/D ratio, IQA, and IL-1, IL-6, TNF-α in lung tissue and BALF, as well as MDA content and MPO activity in lung tissue were significantly increased, PaO2/FiO2 and SOD activity in lung tissue were significantly decreased. The parameters of the XBJ+CPB group were significantly improved, suggesting that Xuebijing pretreatment could improve CPB-induced ALI in rats. The expression of miR-17-5p in lung tissue of the CPB group was significantly down-regulated as compared with sham group (2-ΔΔCt: 0.48±0.13 vs. 1.00±0.11, P < 0.05);while the expression of miR-17-5p in the XBJ group was significantly up-regulated as compared with the CPB group (2-ΔΔCt: 1.37±0.09 vs. 0.48±0.13, P < 0.05), indicating that the improvement of Xuebijing injection on lung injury after CPB might be related to miR-17-5p. ② In vitro experiment: the changes in miR-17-5p expression in each group of AECⅡ cells confirmed in vivo results. After transfection of miR-17-5p mimic, the apoptotic rate and caspase-3 activity of each group were significantly lower than those transfected with negative control, and the decrease was more significant in the XBJ+CPB group [apoptotic rate: (7.37±0.95)% vs. (12.60±1.90)%, caspase-3 (A value): 0.82±0.09 vs. 1.37±0.08, both P < 0.05]. After transfection of miR-17-5p inhibitor, the apoptotic rate and caspase-3 activity of each group were significantly more than those transfected with negative control [in the XBJ+CPB group: apoptotic rate was (16.30±1.86)% vs. (12.60±1.90)%, caspase-3 (A value) was 1.78±0.13 vs. 1.37±0.08, both P < 0.05]. This indicated that the apoptosis of AECⅡ cells cultured in serum after CPB was significantly reduced by miR-17-5p, and further reduced by the pretreatment with Xuebijing. Conclusions Xuebiing injection can reduce the inflammatory reaction and oxidative stress of lung tissue in rats with ALI induced by CPB, and improve oxygenation. The mechanism may be related to up-regulation of miR-17-5p expression in AECⅡ cells and inhibition of apoptosis of AECⅡ cells.

OBJECTIVE To establish the drug use evaluation criteria of human albumin （HA） in the treatment of liver cirrhosis based on weighted technique for order preference by similarity to ideal solution （TOPSIS） method， and to comprehensively evaluate the utilization of HA. METHODS The drug use evaluation criteria of HA was established with reference to guidelines of liver cirrhosis treatment at home and abroad and HA prescription evaluation method， based on the HA instructions. The weighted TOPSIS method was used to evaluate the rationality of HA drug use in inpatients with liver cirrhosis diagnosed by the infection department of our hospital in 2020. RESULTS The established drug use evaluation criteria for HA included 8 evaluation indicators， i.e. baseline examination， indication， contraindication， combined use of drugs， usage and dosage， and so on； the scoring weight was set by consulting experts. According to the evaluation of the closeness degree （Ci） between the HA drug use of 1 068 medical records and the optimal plan and the worst plan in our hospital， the result showed that 133 （12.45%） were evaluated as reasonable medical records （Ci≥0.8）， 576 （53.93%） as basically reasonable medical records （0.6≤Ci＜0.8）， and 359 （33.61%） as unreasonable medical records （Ci＜0.6）. The main unreasonable problems were the absence of baseline examination， non-indication medication， medication for contraindication， etc. CONCLUSIONS The drug use evaluation criteria of HA based on the weighted TOPSIS method can be used to evaluate the rational use of the drug. The drug use of HA in patients with liver cirrhosis in our hospital is reasonable， and we should continue to strengthen the management to reduce or avoid unreasonable drug use.

Objective: To study the relationship between STAT3 gene (rs2293152, rs957970) polymorphism and susceptibility to bronchial asthma in children, and to lay the foundation for search of the candidate pathogenic gene of asthma and early prevention of the disease. Methods: One hundred and thirty hospitalized asthmatic children or out-patients with asthma at Affiliated Hospital of Southwest Medical University from September 2013 to September 2017 were selected as asthmatic group, and at the same time 145 healthy children were selected from the Affiliated Hospital of Southwest Medical University as the healthy control group.Apply sequence specific primers-polymerase chain reaction (SSP-PCR) was applied to detect susceptibility single nucleotide polymorphism (SNP) of STAT3 gene (rs2293152, rs9579700). The statistical differences in distribution between genotype and allele were analyzed between the asthma group and the healthy control group. Results: The distribution frequency of CC genotype of rs2293152 in STAT3 gene was significantly higher in the asthmatic group (19.2%)than that in the healthy control group (9.0%) (χ²=6.065, P=0.014). C allele was significantly higher in asthmatic group (41.9% vs.33.8%) (χ²=3.861, P=0.049). There was no significant difference in the distribution of rs957970 in STAT3 gene between the asthma group and the healthy control group. Conclusions The SNP of rs2293152 in STAT3 gene is associated with susceptibility to asthma in children in South Sichuan.C allele may be the susceptible gene for childhood asthma, while the SNP of rs957970 in STAT3 gene has no significant correlation with childhood asthma in Southern Sichuan.