0.05).But the cell purity and the survival rate by mechanical procedure were significantly higher than that by combination of enzymatic digestion(P

Objective High-throughout sequencing of all plasmid of 206 strains of Klebsiella pneu-moniae using Solexa/Illumina sequencing technology in order to investigate the resistance for plasmids in Klebsiella pneumoniae. Methods Bacterial isolates were obtained over the years 2002-2008. Solexa/Illumi-na sequencing technology was used to sequence both samples (S1 and S2) to a depth of between 10-560 fold coverage. We used SOAP provided by BGI to find SNPS and use velvet package to assemble these sequences and gained some long sequences, and MAQ programs developed in the laboratory were used to annotate SNPs and compare lineage-specific mutations in SHV-ESBLs. Results The Metagenome of plasmid encodes a 13 variety of resistance-related genes with exceptionally high copy numbers, including ABC-type efflux pumps and 4 variety of β-lactamases, SHV-ESBLs is abroad presence. We systematically investigated single nucleo-tide substitutions in plasmids metagenome, and found an amount of nonsynonymous mutations in the SHV-ESBLs genes. Conclusion Probabily in press of positive selection, we can clearly see these nonsynonymous changes predominantly occurred in plasmid SHV-ESBLs genes. And our findings indicate a unspecial low-level resistance contribute to antimicrobial efflux in the metagenome of plasmid in Klebsiella pneumoniae.

Objective To measure the reconstructed cranial CT images,and to clarify the safety range of unilateral nasal transsphenoidal approach for pituitary surgery.Methods 100 normal pituitary cranial CT images were randomly selected,and the three-dimensional reconstruction was performed by using the CT images of perpendicular and parallel to the edge of the two eyes as base line, and the distance and angle in unilateral nasal transsphenoidal approach for pituitary surgery from the sagittal plane in the middle of the nasal meatus and the plane through the tip of the nose and both ends of dorsum sellae were measured,respectively. The angles and distances were compared when grouped the data by gender and age. Results Angle A1 (the angle between the tip of the nose and the tuberculum sellae and saddle back root line in the sagittal plane)in the sagittal plane of the middle nasal meatus was (11.22±1.35)°,95% confidence interval was 8.92°-13.76°degrees;the distance D1(the distance on the line between tuberculum sellae and saddle back root, and the line was formed by the plate contained the angle A1 and sellar floor)was (16.71 ± 2.07)mm,95% confidence interval was 13.11-19.93 mm.Angle A2 (the angle between the tip of the nose and the saddle back ends)which was in the plane through the tip of the nose and both ends of dorsum sellae was (8.91±1.19)°,95% confidence interval was 7.12°-10.72°;the distance D2(the distance on the line between the saddle back ends,and the line was formed by the plate contained the angle A2 and sellar floor)was (14.23±2.09)mm,95% confidence interval was 10.81-17.92 mm. The four parameter data was normally distributed,and there was no significantly statistic difference between different gender and ages (P>0.05).Conclusion The angle of the movement for unilateral nasal transsphenoidal approach for pituitary surgery operation in the sagittal plane in the middle of the nasal meatus should be less than (11.22 ± 1.35)°,and the distance of the movement should be less than (16.71±2.07)mm. The angle of the movement in the plane through the tip of the nose and both ends of dorsum sellae should be less than (8.9 1 ± 1.1 9 )°, and the distance of the movement should be less than (14.23±2.09)mm.

Objective To examine the variation in TLR7 gene copy number of patients with systemic lupus erythematosus (SLE) in Han population,and investigate the relationship between TLR7 gene copy number variations and clinical phenotypes of SLE.Methods Determination of gene copy number of TLR7 was achieved by AccuCopyTM multiple gene copy number detection method in 337 cases of Han SLE patients and 338 healthy controls.According to the clinical phenotype stratification,all cases were divided into lupus nephritis and non-lupus nephritis group,the hematological involvement and non-hematological involvement group,anti-Smith antibody positive and negative group.The data were analyzed by non-parametric rank test.Results Based on sex,there was no significant difference in the variations in TLR7 gene copy number between in female SLE patients and female healthy controls (Z=-1.175,P=0.240).There was also no difference in male group (Z=-1.085,P=0.278).Comparing gene copy numbers variation based on the presence or absence of lupus nephritis,hematological involvement,and anti-Smith antibody,there was no statistical significant difference in female SLE patients(Z=-0.888,P=0.375; Z=-1.085,P=0.278; Z=-0.529,P=0.597).There was no difference in variation in male SLE patients,neither (Z=-0.460,P=0.646; Z=-0.340,P=0.733;Z=-0.158,P=0.874).Conclusion The variations in TLR7 gene copy number are not correlated with SLE and clinical phenotypes of SLE in Han population.

Chloroplast genome sequences have comprehensive application prospects in DNA barcoding and chloroplast engineering in traditional Chinese medicine. The complete chloroplast genome of Magnolia officinalis sequenced by high-throughput pyrosequencing and a sequencing procedure was established. Fourteen contigs were obtained after de nove assembly. The sequencing percent of coverage was 99.99%. The chloroplast genome is 160 183 bp in size, and has a typical quadripartite structure with the large (LSC, 88 210 bp) and small copy (SSC, 18 843 bp) regions separated by two copies of an inverted repeat (IRs, 26 565 bp each). chloroplast genes were successfully annotated, of which 17 genes located in each IR region. The chloroplast genome features in Magnolia officinalis are nearly identical to those from other Magnoliid chloroplast genomes. Phylogenetic analyses were performed based on 81 shared coding-genes for a total of 9 Magnolia samples of 5 closely related species. Results showed that distinguishing among species was generally straightforward at the species and population level. This study confirmed the effectiveness of our chloroplast genome sequencing procedure. The chloroplast genome can provide distinguishing differences to help identify Magnolia officinalis and its closely related plants.

Objective Based on the principles of laser radiation protection, medical metroiogy and photoelectron technology, an automatic verification device and testing technology to provide verification and performance evaluation for laser protective spectacles and equipments which have the various functions in laser protection have been developed and established. Methods The current system comprises laser source, laser measuring instruments, software of computer detection and control and modules of optics and mechanics used in auxiliary equipment. By use of the verification device and the special test-recording method to be designed and studied by us, the measured data can be automatically acquired, processed, recorded, saved and printed and, furthermore, many parameters on protective performance of the measured laser protective spectacles can be tested and given. Results Laser wavelength of the verification apparatus are 1 064 nm and 532 nm, response range of wavelength is from 0.4 m to 1.1 μm, measuring range of laser energy is from 10-8 J to 0.3 J, measuring range of optical density for laser protective spectacles is from 0.1 to 8.0, stability is 0.21%, measuring uncertainty is 5%(k=2). Conclusion The automatic verification device is steady and reliable. The achieved performance indexes accord with the requirement of national standard on laser protection.

Objective To investigate the place reaction of support drainage tube in cholecystojejunostomy and its pathological changes.Methods Eight pigs were randomized into 2 groups.The tube-placing cholecystojeunostomy(n=2)or choledochocholedochostomy(n=2)and non-tube-placing cholecystojejunostomy(n=2)or choledochocholedochostomy(n=2)were performed respectively.All the 8 pigs were sarcrificed 3 months after the operation for pathological examination.Results Gross anatomy catheter group showed anastomotic inflammatory hyperplasia, wall hardening hyperplasia,luminal narrow contracture and stone formation.Microscopic biopsy showed bile duct mucosa in interstitial lymphocytes, plasma cells and other chronic inflammatory cell infiltration, a large number of fibrous tissue hyperplasia.However, these features were not found in pigs without placing the tube.Conclusion The tube can not prevent the stenosis.It might promote the formation of stricture since it can stimulate inflammatory hyperplasia and tissue hardening.

Medicinal animals are important part of Traditional Chinese medicine resources in China. Cytochrome c oxidase subunit I (COI) was selected as the standard DNA barcoding sequence for animal medical materials. In this study, the 51 animal species from 45 animal medical materials in the Chinese Pharmacopoeia were selected and the intra-specific variation and the inter-specific divergence, the barcoding gap, the identification efficiency of their COI sequences were analyzed. The results showed that the inter-specific divergence is higher than intra-specific distance. The barcoding gap existed between inter-specific sequence divergence and intra-specific dis-tance. The identification efficiencies were 100% both at the genus and species level except the Arthropoda. The cluster dendrogram exhibited that different species distinguished from others. Therefore, COI sequence as a bar-code is suitable to identify the species of animal medical materials in Chinese Pharmacopoeia.

Objective:To investigate the predictive value of mucosal vascular pattern (MVP) under narrow-band imaging (NBI) enteroscopy in patients with ulcerative colitis (UC) in clinical remission for histological healing and clinical recurrence.Methods:A total of 142 patients with UC in clinical remission who visited the First Affiliated Hospital of Weifang Medical University from January 2018 to January 2021 were included in the study and underwent colonoscopy. The white light and NBI endoscopic images were collected and biopsies were obtained. The Mayo endoscopic score (MES) was calculated based on white light images, and MVP staging was evaluated based on mucosal vascular patterns under NBI. Nancy index (NI) was used to evaluate histological healing and patients were followed up for 1 year. The Spearman correlation coefficients of MES and MVP with histological healing and recurrence were calculated. The receiver operator characteristic (ROC) curve was plotted and the area under curve (AUC) was applied to evaluate the accuracy of white light and NBI endoscopy for predicting histological healing of UC in clinical remission.Results:According to the MVP criteria, 47 were defined as clear, 63 blurred, and 32 invisible. Spearman correlation analysis showed a significant correlation between MVP under NBI and histological healing ( r=0.549, P<0.001) and a moderate correlation between MES under white light and histological healing ( r=0.462, P<0.001). Spearman correlation analysis showed a moderate correlation between MVP under NBI and clinical recurrence ( r=0.451, P<0.001) and a moderate correlation between MES under white light and clinical recurrence ( r=0.352, P<0.001). AUC of NBI for diagnosing histological healing of UC in clinical remission was 0.809 (95% CI: 0.738-0.879), with a sensitivity of 84.6% (77/91) and specificity of 64.7% (33/51), superior to the white light endoscopy, of which AUC, sensitivity and specificity were 0.763 (95% CI: 0.678-0.848), 81.3% (74/91) and 66.7% (34/51). Conclusion:MVP staging under NBI could predict histological healing of UC patients in clinical remission and is superior to white light endoscopy.

Objective:To observe the clinical effect of acupuncture treatment based on Tiaoshen theory for the patients with functional dyspepsia (FD) and depression with liver-stomach disharmony syndrome. Methods:A total of 76 FD patients from May 2018 to August 2019 were randomly divided into 2 groups with 38 patients in each group. In the routine group, acupoints were selected routinely, and in Tiaoshen group, acupoints were selected by Tiaoshen theory. Both groups were treated for 28 days. The results were evaluated by Nepean Dyspepsia Index (NDI), TCM Syndrome Score and Hamilton Depression Scale-24 (HAMD-24). Results:The total effective rate of both groups was 94.6% (35/37) in Tiaoshen group and 75.0% (27/36) in routine group. There was significant difference between the two groups ( χ2=6.125, P=0.011). The NDI in Tiaoshen group was significantly lower than that of routine group ( t=3.038, P=0.003). The scores of interference domain, control domain, food and beverage domain and sleep disturbance domain in Tiaoshen group were significantly higher than those in routine group ( t=3.096, 2.460, 2.225, 2.732, P<0.05); the TCM Syndrome Score in Tiaoshen group was significantly lower than that of routine group ( t=3.241, P=0.002), and that of HAMD-24 was significantly lower than that of routine group ( t=2.767, P=0.007). Conclusion:Treatment based on Tiaoshen theory can improve the quality of life of FD patients in the fields of interference, control, food and beverage and sleep disturbance, and reduce the patients’depression.