Objective:To express and secrete rBPI m23 bactericidal protein in Pichia pastoris expression system. Methods:pPICZ? synBPI m600 recombinant expression vector was constructed with prepared vectors (pUC18 synBPI 414 and PBV220 BPI m420 ) according to Molecular Cloning Laboratory Manual. The Pichia pastoris GS115 were electroporated with linearized pPICZ? synBPI m600 vectors , positive clones were screened on low salt LB medium with Zeocin , then the positive clones were identified by PCR and Mut phenotype analysis. rBPI m23 was induced to express in BMMY ,purified by SP Sepharose beads , identified by SDS PAGE and Western blot and measured with BCA method. Results:①pPICZ? synBPI m600 expression vector was successfully constructed.②Pichia pastoris GS115 strains integrated with synBPI m600 stably were obtained.③SDS PAGE showed the molecular weight of the recombinant protein was approximate 23 kD,and Western blot confirmed that the protein could bind to the anti BPI antibody specially.④The supernate protein concentration was about 2 9 mg/L.Conclusion:rBPI m23 was expressed and secreted effectively in Pichia pastoris GS115.

Objective To analyze spectral curves and iodine levels in normal adult thyroid by using dual-source CT scanning technique and provide the basis for the differential diagnosis of thyroid space-occupying lesions.Methods A total of 70 subjects were selected for this study,including 38 females and 32 males.They were excluded from thyroid disease by ultrasound examination and had all taken the dual-source CT scan.The image data were transported to the CT workstation,the analysis of iodine levels and spectral curves were completed in theLiver VNCandMonoenergetic modes respectively.The gender differences in ratio iodine content were compared.Results The iodine ratio of thyroid was (1.35±0.19) mg/mlin plain scan,(1.35±0.23) and (1.35±0.21) mg/ml in left lobe and right lobe respectively.There was no statistically significant difference between left and right lobe (P ＞ 0.05),as well as between females and males (P ＞ 0.05).The slopes of the energy spectrum curve in the left and right lobe of the thyroid were 9.98 ± 2.43 and 9.74 ± 1.63,respectively,and 9.88 ± 1.74 and 9.74 ±0.09 for the male and female,respectively.There was no significant difference of slopes of the energy spectrum curve between the left and right lobe,as well as the male and female.The average dose length product of all patients was (208.29 ±21.80) mGy·cm,and the average effective dose was (1.23 ± 0.13) mSv.Conclusions Dual-source dual-energy CT scanning technique have definitive,diagnostic characteristic in the measurement of spectral curves and calculation of the ratio of iodine in normal adult thyroid tissue,and therefore has guidance value in the diagnosis of thyroid space-occupying lesions.

Bartter′s syndrome(BS) is a rare renal tubular diseases and an autosomal recessive hereditary disease. The characteristic findings are hypokalemia, metabolic alkalosis, and raised serum renin and aldosterone levels. Combination of metabolic syndrome with Bartter′s syndrome has not been reported so far. Here in, we report a case in order to call attention to the diagnosis and treatment of typical Bartter′s syndrome combined with metabolic syndrome.

Objective:To explore the effect of autophagy inhibitor 3-methyladenine(3-MA) on camptothecin(CPT)-induced Hela cell apoptosis.Methods:MTT assays were carried out to determine the optimal concentration and time of CPT on Hela cells and the effect of different drugs on Hela cell proliferation activity .After Hela cells were treated with different drugs ,the changes of autophagy marker protein( microtubule-associated protein 1 light chain 3,LC3),p62 and apoptosis-related protein were detected using Western blot and immunofluorescence ( IF) .DAPI ( nuclear ) staining was used to observe cell apoptosis rate .Results: In CPC-treated Hela cells,Hela cell proliferation activity declined dramatically ,and autophagy could be induced to occur .Compared with CPT group ,the cell proliferation activity was lower in CPT combined with 3-MA group,the level of autophagy decreased ,but the apoptosis rate significantly increased.Conclusion:CPT can induce autophagy while inducing Hela cell death .Hela cells chemosensitivity to CPT treatment can be enhanced by 3-MA inhibiting autophagy .

To evaluate the efficacy and safety of Tanreqing Injection, a compound traditional Chinese herbal medicine, for community-acquired pneumonia.

Objective: To investigate the health literacy and its influencing factors among residents in areas out of poverty in Kunming City, so as to provide insights into formulating health education strategies. Methods: A total of 1 916 permanent residents at ages of 15 to 69 years were sampled in three areas out of poverty in Kunming City using the multi-stage stratified random sampling method and probability proportionate to size sampling method from August to October, 2020. Health literacy was investigated using the Questionnaire on the Health Literacy among Chinese Residents, the level of health literacy was analyzed and weighted by the population of the China's Seventh National Population Census. Factors affecting health literacy among residents in areas out of poverty were identified using a multivariable logistic regression model. Results: A total of 1 916 questionnaires were allocated, and 1 908 valid questionnaires were recovered, with an effective recovery rate of 99.58%. The respondents included 997 men (52.25%) and 911 women (47.75%), and had a mean age of (45.58±14.28) years. The level of health literacy was 21.38%. Multivariable logistic regression analysis identified age (15 to 24 years, OR=5.087, 95%CI: 1.573-16.450; 25 to 34 years, OR=6.016, 95%CI: 1.991-18.183; 35 to 44 years, OR=7.526, 95%CI: 2.541-22.289; 45 to 54 years, OR=4.800, 95%CI: 1.640-14.050), educational level (junior high school, OR=5.333, 95%CI: 3.100-9.175; high school/vocational high school/technical secondary school, OR=19.895, 95%CI: 10.418-37.966; college or above, OR=27.580, 95%CI: 12.349-61.597) as factors affecting health literacy among residents in areas out of poverty in Kunming City. Conclusion The level of health literacy is 21.38% among residents in areas out of poverty in Kunming City, and age and educational level are associated factors.

Objective To explore whether different Epstein-Barr virus (EBV) infection status is involved in systemic lupus erythematosus (SLE) pathogenesis through type Ⅰ interferon pathway by observing the EBV antibodies,serum interferon α (IFN-α) level and four type Ⅰ interferon induced gene (ISGs;2'-5' oligoadenylate synthetase-like protein (OASL),myxovirous resistant 1 (MX1),interferon-stimulated gene 15 (ISG15) and lymphocyte antigen 6 complex,locus E (LY6E) expressions in SLE patients and healthy controls.Methods Forty-eight patients with SLE and 36 healthy controls were enrolled into this study.The serum antibodies of EBV capsid antigen-IgG/lgM and EBV nuclear antigen 1-IgG,and serum levels of IFN-α were measured by enzyme linked immunosorbent assay (ELISA) method.Real time reverse transcription polymerase chain reaction was used to test the mRNA levels of OASL,MX1,ISG15 and LY6E in the peripheral blood lymphocytes (2-△△Ct was used to indicate the gene expression).Statistical analysis was performed using t-test or Mann-Whitney U test,Chi square test and Spearman correlation test.Results ① The EBV lytic infection rate (VCA-IgM) in SLE patients was higher than that in healthy controls (40％ vs 11％,x2=5.381,P=0.027).② The serum levels of IFN-α in SLE patients were higher than those in the healthy controls [(206±151) ng/L vs (90± 76) ng/L,t=4.248,P＜0.05],as well as the mRNA levels of OASL,MX1,ISG15 and LY6E [1.8(0.6,5.1) vs 1.2 (0.5,1.4);1.9(1.0,4.4) vs 0.9(0.7,2.5);4.1(1.6,7.8) vs 0.8(0.5,1.7);1.6(0.7,3.3) vs 0.8(0.6,1.2),U=604,560,312,608;P＜0.05,respectively].The mRNA levels of OASL,MX1,LY6E and ISG15 were all positively related to SLE disease activity index (SLEDAI) scores (r=0.319,0.461,0.547,0.484,P＜0.05,respectively).Serum IFN-α levels were elevated in SLE patients with EBV lytic infection than in non-lytic infection patients [(282± 174) ng/L vs (157±114) ng/L,t=2.604,P＜0.05];The mRNA levels of OASL and ISG15 were also elevated in patients with lytic EBV infection [2.0(0.8,7.6) vs 1.2(0.6,3.1);6.2(2.4,15.5) vs 3.3(1.3,6.3),U=377,350,385,354;P＜0.05,respectively].The SLEDAI scores in patients with EBV lyric infection were higher than in patients with non-lyric infection (16±4 vs 12±8,P＜0.05).Conclusion EBV infection may be involved in the pathogenesis of SLE by activating the type Ⅰ interferon pathway.

ObjectiveTo investigate the effects of autologous bone marrow-derived mesenchymal stem cells (MSCs)transplantation on acute myocardial infarction in swine models using MRI. MethodsFourteen Chinese mini-pigs(27±3 kg)were divided into control group(n=7)and transplantation group(n=7).Acute myocardial infarction(AMI)model was made by occlusion of the left anterior descending coronary artery for 90 minutes,and then 10 ml autologous MSCs(3 × 106 cell/ml)were injected into LAD by over-wire-balloon catheter after one week. MRl was performed to assess the cardiac function and myocardial perfusion 1 week after AMI and 6 weeks after transplantation.The implanted cells in vitro were analyzed by immunofluorescence.ResuitsThe left ventricular ejection fraction(LVEF)in transplantation group was increased from(42.7 ±7.5)%to(50.1±10.1)%,which was significantly different from that in control group(P＜0.01).In addition,the dyskinetic segments in infarcted region and the infareted area were decreased by 4 and 3.2 cm2 respectively(P＜0.01),and the left ventricular weight index was increased by 4.1 g/m2 in transplantation group(P＜0.05)compared with control group.The DAPI-labeled cells in infarcted and peri-infarcted region indicated the survived MSCs.Immunofluoreseence also confirmed that those cells expressed cardiomyocyte-specific troponin T,connexin 43 and vessel-specific smooth muscle actin.Capillary density in both infarcted and peri-infarcted region were higher in transplantation group than the control group(P＜0.01).Conclusion MRI is a reliable imaging method for assessing the effects of stem cell transplantation in acute myocardial infartion of swine models.

Objective To improve the arthroscopic posterior cruciate ligament (PCL) reconstruction using tibial Inlay technique. Methods The special arthroscopic device and related fixation technique were designed. Five cadaveric knees were used to simulate the process of arthroscopic posterior cruciate ligament reconstruction using tibial Inlay technique. The knees were cut open to observe whether the outlet of the tibial tunnel shape and location met the design requirements. Thirty normal MRI films were measured to identify tunnel angle and localizer angle. Results The inner outlet of tunnel was conical shape(14 mm×7 mm×15 mm) and the outer outlet was cylinder-shaped (a diameter of 7 mm). The tibial drill was designed into a split structure and could be assembled in vitro. According to the data obtained from MRI films, the angle between the plane of posterior cruciate ligament and horizontal place was 36°-47°, and the localizer was fixed at 50°.The achilles tendon was used as implant and the allogft bones were designed into conical shape to fit the inner outlet of tunnel. The other end of implant to the proximal tibia was fixed with button plate. All reconstruction operations were performed under arthroscopy. The outcomes of procedure were satisfactory. There were no vascular or peripheral nerve injuries in the cadaveric knees The tunnel position was accurate and the shape of tunnel had met the design requirements. Conclusion Our results imply that improved arthroscopic of posterior cruciate ligament using tibial Inlay technique is simple, accurate, rapid and stable fixation.

Objective To evaluate the protection of combined clostridium butyricum and bifidobac-terium living powders for antibiotic-associated diarrhea ( AAD ) with all kinds of infections in hospitalized children,and to compare the therapeutic effect with saccharomyces boulardii. Methods This study was a prospective,randomized case-control clinical trial which collected the data of the hospitalized children with all kinds of infections in Pediactric Department of Shengjing Hospital of China Medical University between May 2011 to May 2012. A total of 552 cases were enrolled and 480 cases completed the study. A total of 240 chil-dren were in experimental group,80 cases received combined clostridium butyricum and bifidobacterium liv-ing powders 840 mg per time,twice a day and the other 160 cases received saccharomyces boulardii 250 mg per time,twice a day,for one week; the control group took none of probiltics. Two groups received routine antibiotic therapy. Everyday′s defecate frequency was recorded, the traits of excrement according to bristol stool assessment scale were evaluated,the incidence of diarrhea and drug related adverse reactions were coun-ted. Results During the studied 7 days,the AAD incidence was 4. 2%(10/240) in experimental group and 20. 4%(49/240) in control group,there was significant difference between two groups. The risk of AAD in experimental group decreased 58. 5%. Compared to saccharomyces boulardii,combined clostridium butyricum and bifidobacterium living powders decreased 38. 2% (RR=0. 728, 95%CI 0. 257~0. 784, P=0. 009). Compared to control group,the average defecate frequency decreased in experimental group,diarrhea duration contracted,there was statistic difference between two groups ( P<0. 01 ) . No drug related adverse reactions happened during the trial. Conclusion Both combined clostridium butyricum and bifidobacterium living powders and saccharomyces boulardii could effectively reduce the risk of AAD in hospitalized children with bacterial infection,relieve diarrhea symptoms,short the duration of diarrhea,and did not find the adverse reac-tions. Combined clostridium butyricum and bifidobacterium living powders and saccharomyces boulardii had the same protective effect for AAD of northern China children.