Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

Objective:To explore the clinical value on application of laparoscopic ultrasonography (Lap US) in the adnexal operation. Methods:Eleven patients including 7 cases of tubal pregnancy, 3 cases of teratoma of ovary and 1 case of endometrial cyst of ovary were examined by LapUS, then operated with laparoscopy.Results:The results showed that the modality presented is a big progress over the traditional operative management for adnexal diseas. Conclusions:It is a good approach for micro-surgery in the adnexal operation by laparoscopy.

Objective To investigate the epidemiological characteristics and mortality risk factors of severe sepsis patients admitted into intensive care unit (ICU).Methods The clinical data of 419 severe sepsis patients admitted to an adult ICU of Beijing Friendship Hospital from January 2009 to December 2012 were retrospectively analyzed and evaluated.Logistic regression analysis was employed to identify independent risk factors for the death of patients with severe sepsis during ICU stay.Results Overall ICU mortality was 43.9％ (184/419),and the respiratory tract was the most common site of infection (50.8％),followed by infection of the gastrointestinal tract and abdominal cavity (27.8％),and hematogenous infection (4.3％).Gram-negative bacteria were the most common pathogens [46.0％ (214/465)].Iatrogenic infections accounted for 53.7％ (225/419) of the enrolled patients.Logistic regression analysis showed that age [odds ratio (OR)=0.003,95％ confidence interval (95％CI) 1.015-1.056,P=0.O00],vasoactive drug application (OR=3.251,95％CI 1.562-6.768,P=0.002),failure of 3 or more organs (OR=2.452,95％ CI 1.015-5.924,P=0.046),and iatrogenic infection (OR =1.775,95％ CI 0.981-3.221,P=0.046) were independent risk factors for ICU mortality.Conclusions Severe sepsis is a common cause of ICU admission.Patients with risk factors for high mortality should be carefully monitored,and aggressive treatment should be administered.

Objective To study the liver protection of total celosins (CES) in different liver damage models .Method A-cute liver damage mouse model ,chronic liver damage rat model and immune liver damage mouse model were established for this study .Liver function tests ,liver lipid peroxidation indicators and liver pathology were observed .Results CES of each dose group can lower serum ALT and AST in different degrees ,can significantly reduce the content of MDA in liver homogenate , while can increase SOD activity .Pathological examination showed that animals administered significantly reduced the degree of liver cell damage .Conclusion CES have protective effects on acute liver damage induced by CCl 4 ,chronic liver damage induced by CCl4 and immunological liver damage induced by concanavalin A .Its hepatoprotective effect may be related to its antioxidant effects .

Objective To analyze the characteristics of change in plasma cholesterol level in patients with sepsis, and to explore its relationship with prognosis and its clinical significance. Methods A retrospective analysis was conducted. 568 patients with sepsis admitted to Department of Critical Care Medicine of Beijing Friendship Hospital Affiliated to Capital Medical University from August 2013 to August 2015 were enrolled, and 475 patients without sepsis hospitalized in the same period served as the control. The basic clinical data of the two groups were collected, and the results of blood fat and biochemical parameters were compared. The patients with sepsis were divided into death group and survival group, and risk factors influencing the prognosis of patients with sepsis were analyzed with multivariate logistic model regression analysis. Results Compared with non-sepsis patients, the levels of plasma total cholesterol (TC) and high density lipoprotein cholesterol (HDL-C) in the patients with sepsis were significantly lower [TC (mmol/L): 2.5±1.2 vs. 3.4±1.4, t = 4.274, P = 0.021; HDL-C (mmol/L): 1.6±0.9 vs. 2.5±0.8, t = 3.413, P = 0.018], and that of low density lipoprotein cholesterol (LDL-C) showed no statistically significant difference (mmol/L: 1.9±0.9 vs. 2.1±0.9, t = 0.749, P = 0.614). In the patients with sepsis, the patients in death group (n = 227) were older than those of the survival group (n = 341, years: 74.3±15.5 vs. 65.5±17.5, t = 4.037, P = 0.012), serum creatinine (SCr) was higher than that of survival group (μmol/L: 251.0±115.6 vs. 167.4±108.7, t = 3.254, P = 0.023), the levels of plasma TC, HDL-C and LDL-C were significantly lower than those of survival group [TC (mmol/L): 2.2±1.6 vs. 2.9±1.1, t = 3.057, P = 0.023; HDL-C (mmol/L): 1.4±0.8 vs. 1.9±0.8, t = 4.692, P = 0.016; LDL-C (mmol/L): 1.7±0.7 vs. 2.0±0.8, t = 2.541, P = 0.038]; there was no significant difference in the proportion of male, body mass index (BMI), based disease, intensive care unit (ICU) hospitalization time, the severity of the disease and other biochemical indexes between two groups. With single factor analysis with indicators of statistical significance as a covariate into binary logistic regression equation, the results show that age was the risk factor of death in patients with sepsis [odds ratio (OR) = 1.024, 95% confidence interval (95%CI) = 1.010-1.048, P = 0.009], and TC was the protective factor on the prognosis of patients with sepsis (OR = 0.747, 95%CI = 0.682-0.811, P = 0.013). Conclusions Plasma cholesterol levels in patients with sepsis were significantly lowered, and the levels in death group was significantly lower than that in the survival group. TC may be used as a clinical indicator to assess the outcome of patients with sepsis.

AIM: To study the cytoskeleton of mesench ymal stem cells (MSCs), the ultrastructure and function relationship by using atomic force microscope (AFM). METHODS: The ultrastructures and morphological feature of MSCs c ultured for 1 d and 5 d were studied by AFM. RESULTS: The special structures that possess peculiar morphologi cal characteristic of MSCs such as cytoskeleton, pseudopod, microfilament etc we re identified by AFM, and these special structures are difficult to observe unde r electronic microscopy or other conventional optical microscopy. CONCLUSIONS: AFM is a powerful tool to study ultrastructures, mo rphological features, and cytoskeleton of stem cells in near physiological condi t ions. Its application prospect in cellular biology is extensive. The special cyt oskeleton and other structures of MSCs observed above may represent the structur al base of multi-differentiation potential of MSCs.

Objective To study on the difference of plasma renin activity ( PRA),angiotensin Ⅱ (Ang Ⅱ ),and aldosterone levels in patients with essential hypertension (EH) or primary aldosteronism (PA) or pheochromocytoma (PHEO),and to analyze the sensitivity and specificity on the diagnosis of PA among patients with hypertension with aldosterone/PRA ratio (ARR).Methods The plasma aldosterone,Ang Ⅱ and PRA concentrations in supine and upright positions were measured by radioimmunoassay from 413 patients including idiopathic hyperaldosteronism (IHA,n =111 ),aldosterone-producing adenoma (APA,n=l18),PHEO (n=98) and EH (n=86).ARR was calculated.Results Plasma aldosterone concentrations in both of supine and upright positions in PHEO group [ 374 (294,465 ) pmol/L and 629 (449,997) pmol/L] and PA group [471 (346,632) pmol/L and 673(499,825) pmol/L] were higher than those in EH group [ 277 (224,332) pmol/L and 427 (341,501 ) pmol/L ] (P ＜ 0.01 ).They were also higher in APA group [576 (416,731 ) pmol/L and 726 (554,906 )pmol/L ] than those in IHA group [399(313,504) pmol/L and 609(485,776)pmol/L ] (P ＜0.01).Ang Ⅱ levels in both positions were lower in PA group [43.2(26.4,74.4) ng/L and 60.1(38.5,103.6) ng/L] than in EH group [56.7 (43.3,78.9) ng/L and 84.3(61.3,108.4) ng/L] or PHEO group [54.3(29.9,101.5) ng/L and 102.8 (49.9,167.0) ng/L] (all P values ＜ 0.01 ),and there was no difference between IHA and APA group (P ＞ 0.05 ).The PRA level in both positions of each group were PHEO group [ 0.3 (0.2,1.0) μg ·L-1 · h-1 and 1.4(0.6,3.4) μg · L-1 · h-1] ＞EH group [0.2(0.1,0.4)μg · L-1 · h-1 and 0.6(0.4,1.0)μg· L-1 ·h-1] (P＜0.01) ＞PAgroup [0.1(0.1,0.1)μg· L-1 · h-1 and 0.2(0.1,0.3)μg·L-1 · h-1] (P＜0.01),and APA group [0.1(0.1,0.1)μg · L-1 · h-1 and0.1(0.1,0.3)μg · L-1 ·h - 1 ] ＜ IHA group [ 0.1 ( 0.1,0.2 ) μg · L - 1 · h - 1 and 0.2 (0.1,0.3 ) μg · L-1 · h - 1 ] ( supine P ＜0.01 ; upright P ＜ 0.05 ).APA was divided into 2 types with renin-Ang Ⅱ -responsive APA ( n =26) and unresponsive APA (n =92).The plasma aldosterone concentration was lower in supine position but higher in upright position in renin-Ang Ⅱ-responsive APA than in unresponsive APA patients.ARR in upright was higher in PA group ( P ＜ 0.01 ) but lower in PHEO group ( P ＜ 0.05 ) compared with EH.ARR was higher in APA than in IHA (P ＜0.01 ).The sensitivity and specificity of ARR as 40 (aldosterone unit:ng/dl;PRA unit:μg · L-1 · h-1; its value should multiply 27.7 when transferred to pmol/L,simili) were 93％ and 76％,respectively.Conclusion The levels of PRA,Ang Ⅱ and aldosterone from patients with EH,PA and PHEO are significant different.ARR as 40 in upright position could be used for PA screening cutoff point.

Objective A state quo survey of laboratory animal resource in Guangdong province is performed to provide reference data for government management decision-making and market assessment of laboratory animals. Methods We used questionnaires focusing on the laboratory animal facilities with authorization by Guangdong Province Government, which mainly included the production and use of laboratory animals,the qualification of employees and the facilities space. Results The total production and use of laboratory animals(except for the eggs)had been increasing in the last four years. 1.57 million laboratory animals were produced and 0.754 million laboratory animals were used in 2016. There were 2352 employees,roughly the same as in 2015. The facilities space for breeding was 121008 m2,and for animal experiment was 73470 m2,which were rising in the past three years. Conclusions In order to reinforce the industry development of laboratory animals in Guangdong province,some suggestions were given in our study,such as facilitating the application of superiority resource including non-human primate and aquatic laboratory animals,supporting the standardization production of several scarce mice and rats, improving relevant employees' overall level and constructing laboratory animal facilities sharing platform.

Experimental autoimmune encephalomyelitis (EAE) is an autoimmune disease of the central nervous system (CNS); it serves as a model for the human multiple sclerosis (MS). In mice, EAE is mediated by T cells specific for various myelin basic proteins which migrate from the periphery to the CNS. In search of a way to prevent the induction and progression of EAE, we observed the effects of recombinant immunotoxin IP10-DT390 on blocking or eliminating the active T cells in the EAE model. In this paper is presented an experimental gene therapy-based model in which the mice were made resistant to EAE induction by plasmid DNA encoding recombinant immunotoxin that was injected into the leg muscles of mice. The new immuno-biological construct could selectively impair autoreactive T-cell homing while the duration of clinical signs is shorter, and the new construct would not affect other components of the immune response. These data demonstrated the effectiveness of the constructs in the treatment of EAE and suggested its usefulness in the treatment of other autoimmune diseases.
Animals ; Chemokine CXCL10 ; biosynthesis ; genetics ; therapeutic use ; Diphtheria Toxin ; biosynthesis ; genetics ; therapeutic use ; Encephalomyelitis, Autoimmune, Experimental ; immunology ; pathology ; therapy ; Female ; Genetic Therapy ; Immunoglobulin Fragments ; biosynthesis ; genetics ; therapeutic use ; Immunotoxins ; genetics ; metabolism ; therapeutic use ; Mice ; Mice, Inbred C57BL ; Receptors, CXCR3 ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; therapeutic use ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; T-Lymphocytes ; immunology ; Transfection

Objective To evaluate the value of MR angiography in thrombolytic therapy of acute ischemic stroke. Methods According to inclusion criteria, 65 patients who also having large vessel occlusion were selected, and they were performed rt-PA treatment (38 patients) and routine treatment (27 patients) within 3-6 hours of onset of symptoms, respectively. Mann-Whitney U test and chi square test were performed to compare the clinical and MR imaging baseline index and the clinical outcome between the two groups respectively. Clinical outcome was assessed after 3 months using a dichotomized modified Rankin scale score.Data were also compared with the combined analysis of the ATLANTIS, ECASS, NINDS rt-PA trials. Resets The difference of clinical outcome in 3 months between the two groups was significant (P < 0. 05) and the median of the two group was 1 and 3, respectively. The ratio of favorable outcome (mRS 0-1) in the two groups was 52. 6% (20/38) and 33.3% (9/27), respectively. Conclusion MR angiography plays an important role in thrombolytic therapy of acute ischemic stroke and it should be used to consummate the conventional inclusion criteria, the patients with large vessel occlusion should be treated by rt-PA.