Objective To analyze the platelet activating factor acetylhydrolase ( PAF-AH) activity of a gene product encoded by LA2144 gene of Leptospira interrogans ( L. interrogans) , to investigate the ex-pression and secretion of LA2144 protein in various cell cultures and to further understand its function in in-ducing internal hemorrhage in an animal model. Methods The DNA sample containing LA2144 gene was extracted from L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai and used as the template for gene cloning by PCR. The LA2144 gene without the signal sequence coding region was amplified by PCR and inserted into a prokaryotic expression construct for the protein expression. The expressed recombinant protein, rLep-PAF-AH, was purified by Ni-NTA affinity chromatography. Spectrophotometry was used to measure the hydrolytic activity, hydrolytic efficiency, Km and Kcat values of the rLep-PAF-AH protein in hydrolyzing PAF substrate. Real-time fluorescent quantitative RT-PCR ( qRT-PCR) and Western blot assay were performed to measure the expression of LA2144 gene at mRNA and protein levels in human umbilical vein endothelial cells (HUVEC), human monocytes (THP-1) and murine macrophages (J774A. 1) with L. interrogans strain Lai infection, respectively. Each syrian hamster was intravenously injected with 100 μg of LPS-free rLep-PAF-AH for two times. Hemorrhage in the lungs, livers and kidneys were observed in three days after the injection. Results The constructed prokaryotic expression system for LA2144 gene of L. inter-rogans strain Lai could highly express the rLep-PAF-AH upon the induction of IPTG. The purified rLep-PAF-AH showed high purity with a single protein band in gel as indicated by SDS-PAGE. The efficiency of 5 μg of rLep-PAF-AH in hydrolyzing PAF substrate was 26. 6 U/L with a Km value of 82. 79 μmol/L and a Kcat value of 0. 24 S-1 . The expression of Lep-PAF-AH at mRNA level in HUVEC, THP-1 and J774A. 1 cells were significantly elevated after co-culture with L. interrogans strain Lai for 1 or 2 hours (P<0. 05). A large amount of Lep-PAF-AH were detected in the supernatants from co-cultures of L. interrogans strain Lai with the three cell lines, but not from the culture of the spirochete in EMJH medium. The signs of hemor-rhage were observed in the lung of hamsters injected with rLep-PAF-AH, but not in tissue samples from liver and kidney. Conclusion The LA2144 gene product was characterized by a stronger PAF-AH activity. The expression of LA2144 gene at mRNA and protein levels in various cell lines were enhanced during L. interro-gans infection. Moreover, the rLep-PAF-AH could induce the pulmonary hemorrhage in hamsters. This stud-y indicated that the protein encoded by LA2144 gene was an important virulence factor causing hemorrhage in hosts during L. interrogans infection.

Objective To explore the prevalence and management status of asthma in downtown of Qingdao City.Method A stratified-cluster-disproportional random sampling survey was conducted with uniform procedure and questionnaire.Results The overall incidence of asthma in Qingdao City was 3.12％ (188/6026).The 2 most frequently risk factors of asthma were the allergic history and the asthma heredity (OR =3.562,2.381,P ＜ 0.05 ).In 188 diagnosed asthma patients,only 65 (34.5％)asthma patients accepted the guideline therapy,and 51 (27.1％)of them achieved well-controlled or total control.The 2 common reasons for the poor compliance were fear of adverse drug reaciion and feeling of ineffectiveness of inhaled corticosteroids.Conclusion The population in urban area of Qingdao City is suffered with high prevalence of asthma,poor compliance of guideline treatment and terrible management status.

Objective To investigate magnetic resonance imaging(MRI) characteristics of autoimmune pancreatitis(AIP) in middle-aged and elderly patients. Methods MRI were performed in 17 middle-aged and elderly patients with AlP proved histopathologically or clinically. All clinical and MRI data were reviewed. Results Of 17 cases, MRI showed diffuse enlargement of the pancreas in 13 cases, and focal lesions in 4 cases. Swelled pancreas was hypointense on T1-weighted images, mildly hyperintense on T2-weighted images, decreased enhancement on artery phase of dynamic imaging, and moderate enhancement on delayed phase images. Capsule-like enhanced rim was found around swelled pancreas in 13 patients. Stricture of distal common bile duct was present in 15 patients, and diffuse irregular narrowing of the main pancreatic duct was showed in 5 cases. The mean apparent diffusion coefficient(ADC) value of AIP was 1.64×10~(-3) mm~2/s in 13 cases. Conclusions AIP occurs more often in middle-aged and elderly people. AIP shows some characterized MRI findings including swelled pancreas looking like banana or sausage, delayed enhanced capsule-like rim around the lesion and diffuse irregular narrowing of pancreatic duct. MRI will play an important role not only in the diagnosis of AIP, but also in evaluating curative effect of AIP.

Objective To investigate the role of DWI in differentiating autoimmune pancreatitis ( AIP) from pancreatic cancer ( PC) , and in the therapeutic effect evaluation of AIP.Methods DWI data of 26 cases with AIP , 29 cases with PC and 30 cases with normal pancreas ( NP ) were analyzed retrospectively.The distribution type and signal feature of lesions in cases with AIP or PC were evaluated by Chi-squared test.ADC values were measured and compared among 3 groups by Kruskal-Wallis test.ADC values of AIP and PC were analyzed by using ROC curve to determine the optimal threshold and diagnostic efficiency.ADC values were compared in AIP ( n=15 ) before and after steroid therapy by paired t test.Results Diffuse lesions were detected in 21 cases with AIP and 3 cases with PC, while focal lesions in 5 cases with AIP and 26 cases with PC (χ2 =27.64, P<0.01).On DWI, most of AIP (n=19) and PC (n=24) showed hyper-intense signal, while a few of AIP (n=7) and PC (n =5) showed iso-intense signal (χ2 =0.75, P>0.05).The median ADC values of AIP, PC and NP were 1.15 ×10 -3,1.35 × 10 -3 ,1.59 ×10-3 mm2/s, respectively; and the difference was statistically significant ( H=45.60, P <0.01).ROC analysis yielded an optimal ADC cutoff value of 1.255 ×10 -3 mm2/s (80.8% sensitivity, 79.3%specificity and 0.871 area under curve for the diagnosis of AIP ).ADC values of AIP ( n=15) were markedly increased from the baseline (1.10 ±0.19) ×10 -3 to (1.57 ±0.12) ×10 -3 mm2/s after steroid therapy (t=-10.14, P<0.01).Conclusions DWI may be useful for diagnosing and evaluating the effect of steroid therapy in AIP.ADC values of AIP were significantly lower than those of pancreatic cancer and normal pancreas.After steroid therapy , ADC values were markedly increased in AIP.

Objective To explore the overall situation and development trend of the hierarchical diagnosis,to provide reference for further research direction.Methods Following databases,such as CBM,CNKI,Wanfang Database and VIP,were used to search for publications on this topic from the establishment of database to September 30th,2016.Bibliometric method was used to analyze these literatures,for instance periodical distribution,years,authors and regions;key words were taken into consideration.Results 243 literatures in total were included,the a number of publication is keep increasing every year.These papers were published in 91 kinds of journals,43.2 ％ of which were statis tical source journals.There were 221 first authors from 149 institutions distributed in 26 provinces and municipalities.There were only 18 core authors.The active regions of research were Beijing,Hubei,Sichuan and Shanghai.The research content mainly focused on two-way referral,utilization of medical service and medical insurance.Conclusions The quality of papers is low,research authors are widely distributed,the research strength is not strong enough.The research should be strengthened in extent and depth to transform the research outcomes to guide the actual practice.

Objective To have SD rats inhaled with different drugs,and observe their lung pathological change of lungs through light microscopy,in order to evaluate the safety of different drugs inhaled by natural rats. Methods A total of 40 rats were randomly divided into 8 groups,and every group had 5 rats,including blank control groups,9 g/L saline group,Salbutamol group,Dingchuantang group,Shuanghuanglian group,Centamicin group,Danshen group,Silicon dioxide group,twice a day,last 56 days totally. Then,blood and bronchoalveolar lavage fluid were collected and analyzed for cell count,percent of each type of cell,to measure the severity of the inflammation. Additionally,histopathology re-vealed the lungˊs pathological change and the number of dust cell;while immunohistochemistry revealed CD163 respon-ding. Results (1)White blood cell count:blank control group(3. 96 ± 0. 36)×109/L,9 g/L saline group(4. 66 ± 0. 58)×109/L,Salbutamol group(4. 06 ± 0. 86)×109/L,Dingchuantang group(8. 98 ± 1. 08)×109/L,Shuanghuang-lian group(7. 10 ± 0. 88)×109/L,Centamicin group(6. 14 ± 0. 89)×109/L,Danshen group(9. 84 ± 2. 33)×109/L, Silicon dioxide group(8. 99 ± 2. 48)×109/L,and comparative analysis of the 8 groups had significant difference(F＝14. 530,P<0. 05);the differences among blank control group,9 g/L saline group and Salbutamol group were not sig- nificant(all P>0. 05). White cell count in BALF:blank control group(2. 16 ± 1. 04)×109/L,9 g/L saline group (3. 94 ± 0. 67)×109/L,Salbutamol group(4. 36 ± 1. 15)×109/L,Dingchuantang group(14. 58 ± 2. 93)×109/L, Shuanghuanglian group(19. 68 ± 6. 29)×109/L,Gentamicin group(11. 74 ± 1. 03)×109/L,Danshen group(44. 75 ± 10. 8)×109/L,Silicon dioxide group(53. 54 ± 14. 25)×109/L,and comparative analysis of the 8 groups had signifi-cant difference(F＝40. 616,P<0. 05);the differences among blank control group,9 g/L saline group and Salbutamol group were not significant(all P>0. 05). Lymphocyte count in BALF:blank control group(18. 70 ± 9. 00)×108/L, 9 g/L saline group( 36. 01 ± 5. 99 )×108/L,Salbutamol group( 38. 95 ± 11. 69 )×108/L,Dingchuantang group (132. 70 ± 26. 94)×108/L,Shuanghuanglian group(173. 56 ± 57. 6)×108/L,Gentamicin group(106. 60 ± 16. 76)× 108/L,Danshen group(340. 63 ± 70. 97)×108/L,Silicon dioxide group(495. 63 ± 131. 95)×108/L,and comparative analysis of the 8 groups had significant difference(F＝41. 980,P<0. 05);the differences among blank control group, 9 g/L saline group and Salbutamol group were not significant(all P>0. 05).(2)Number of lung dust cell count in 10 sight of high light microscopy:blank control group 12/10 HP,9 g/L saline group 26/10 HP,Salbutamol group 17/10 HP,Dingchuantang group 262/10 HP,Shuanghuanglian group 133/10 HP,Gentamicin group 109/10 HP,Danshen group 96/10 HP,Silicon dioxide group 315/10 HP,and comparative analysis of the 8 groups had significant difference (F＝69. 915,P<0. 05);the differences among blank control group,9 g/L saline group and Salbutamol group were not significant(all P>0. 05).(3)Hematoxylin-eosin staining of lung:blank control group,9 g/L saline group and Sal-butamol group had no pathological change in the lung,but Salbutamol group,Dingchuantang group,Shuanghuanglian group,Gentamicin group,Danshen group and Silicon dioxide group had pathological changes in different degrees.(4) Immunohistochemistry of CD163 responding:blank control group,9 g/L saline group and Salbutamol group had negative expression,Salbutamol group,Dingchuantang group,Shuanghuanglian group,Gentamicin group,Danshen group and Sili-con dioxide group had positive expression in different degrees. Conclusions 9 g/L saline,salbutamol for atomized inhalation does not cause lung tissue damage;Long-term use of non-atomized drugs in atomization can cause lung tissue injury in SD rats,and the severity varies with specific drugs.

ncreatic enlargement, MRI is comparable to CT.

OBJECTIVETo discuss the genetic diagnosis of congenital adrenal hyperplasia (CAH) and investigate the resource of gene mutations in CAH.

METHODEnzymatic methods with restriction endonucleases that specifically recognized the mutation sites were used to detect the gene mutations in patients with CAH and their relatives. Polymerase chain reaction and direct sequencing were used to identify the mutations in 21-hydroxylase gene, and short tandem repeat (STR) typing was used to determine the sources of the mutations.

RESULTSOne CAH patient had two known mutations in 21-hydroxylase gene, namely the I2g and I172N mutations. The former mutation was inherited from the biological mother and the latter was not inherited.

CONCLUSIONThe 9 common mutations of CAH are also the hotspots for new mutations.

Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Gene Deletion ; Genotype ; Humans ; Mutation ; Point Mutation ; Polymerase Chain Reaction ; Steroid 21-Hydroxylase ; genetics

Focus on the defects of the extraction method of velocity pulsated signals in the velocity spectrum image of pulsed Doppler ultrasounds (such as the limited data, great amount of work and low reproducibility), we combine the earlier research results and the basic rules of the velocity spectrum image, and employing the visual basics plateau, to design and develop a software to extract the velocity signals from the spectrum image automatically. Compare the mean velocity and RNS between the PIV and the PDU resulting from the method, the correlations are r = 0.93 and r = 0.78 respectively. The study reveals the software increase the amount of valid data, decrease the amount of work and increase reproducibility of the signals extraction. The method may be an effective way to analyse the velocity spectrum image of PDU in clinic.
Arteries ; diagnostic imaging ; Blood Flow Velocity ; physiology ; Humans ; Pulsatile Flow ; Signal Processing, Computer-Assisted ; Software ; Ultrasonography, Doppler, Pulsed ; methods

Objective Investigate the effect of gestational diabetes mellitus (GDM)on the neonatal metabolism and prognosis.Methods A total of 265 singleton pregnancy GDM patients were collected as the observation group,while 260 cases of healthy singleton pregnant women were chosen as the control group.Detected the levels of adiponectin,insulin,C-peptide,glycated hemoglobin and blood glucose,compared the prognosis of two groups.Results Compared with the control group,adiponcetin and blood glucose in 2 hours after birth in observation group were lower,insulin,C-peptide and glycated hemoglobin were higher,the number of cesarean section was more,the difference was statically significant (P＜0.05).The incidence of low birth weight infant,macrosomia,neonatal asphyxia,neonatal hypoglycemia,neonatal hyperbilirubinemia,neonatal pneumonia,fetal death and newborn teratogenesis in the observation group was higher than that in the control group,the difference was statically significant (P＜0.05).Conclusion GDM could cause metabolic disorder and affect fetal growth and development,leading to poor prognosis.