Objective To determine the function of conserved amino acids in the fusion-promoting domain of Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein,clearly understanding mechanism of cell fusion.MethodsUsing a PCR-based site-directed mutagenesis method and the method of homology recombination occurred in vivo to change six conservative amino acids into alanine respectively.Wild type (WT) and all mutant HN proteins were exepressed in BHK-21 cells by the vacciniaT7 RNA polymerase expression system.The amount of each HN protein at the cell surface was determined by fluorescence-activated cell sorter (FACS).Cell fusion efficiency,hemadsorption activity (or receptor binding activity) and neuraminidase activity were determined.Results There was no statistic difference of cell surface expression among WT and each mutant HN protein ( P＜0.05 ).Cell fusion efficiency of each mutant protein decreased to some extent,especially 1103A decreased to 14.2％ in head.Hemadsorption activity of mutant proteins were reduced in different extent,the maximum reduction of which was also 1103A,28.2％ of wt NDV HN.There was different neuraminidase activity among each mutant HN protein.L74A increased slightly to 118.6％.L110A decreased most to 5.2％.I103A decreased second most to 5.7％.Conclusion Conserved amino acids in fusion-promoting domain of NDV HN played an important role in cell fusion.I103 was identified as a key amino acid in this domain.

OBJECTIVESTo investigate incidence of perioperative cardiovascular events, to analyze related risk factors for the patients undergoing intraperitoneal surgery.

METHODSThe data of 1079 patients who underwent intraperitoneal surgery (exclude laparoscope surgery) from July 2007 to June 2008 was reviewed and analyzed.

RESULTSFor the patients undergoing intraperitoneal surgery, the incidence of major cardiovascular events was 3.99% (43/1079), all-cause mortality was 1.58% (17/1079). The independent risk factors of major cardiovascular events were age ≥ 60 years, history of coronary heart disease, cardiac insufficiency, arrhythmia, chronic obstructive pulmonary disease, estimated glomerular filtration rate (eGFR) < 60 ml/(min·1.73 m(2)), emergency surgery and duration of surgery > 2.82 h (OR = 2.68 to 5.19, P = 0.001 to 0.031).

CONCLUSIONSThe cardiac risk of intraperitoneal surgery is 3.99%. The risk of cardiac complications should be evaluated in elderly patients and those with ischaemic heart disease, chronic obstructive pulmonary disease, and renal disease, more specifically, when emergent or long duration major surgeries are needed.

Abdomen ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cardiovascular Diseases ; epidemiology ; mortality ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; mortality ; Risk Factors ; Young Adult

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification

Objective:To explore the application of carotid ultrasonography in detecting carotid artery lesions of patients with ischemic cerebrovascular disease(ICVD).Methods:Carotid ultrasonography and vascular reactivity(VR)were performed on 82 patients with ICVD and 40 health controls with Color Doppler Ultrasound.The ultrasonic indexes were statistically analyzed. The intima-media thickness(IMT)and the plaque score,the plaque total area and the cerebral blood flow volume of two groups were compared with each other.Results:Vessel diameter (VD),IMT,peak systolic velocity (PSV),end-diastolic velocity (EDV),mean flow velocity(MFV)in patients with severe stenosis of internal carotid artery and common carotid artery in ICVD group were significantly higher than those in control group,and the differences were statistically significant(P <0.01).VR in patients without stenosis and with mild,moderate and severe stenosis was lower than that in the control group(P <0.05).In ICVD group,IMT,carotid plaque score and plaque total area were significantly higher than those in the control group(P <0.01).Conclusions:The detection on degree of carotid lesions in patients with ICVD via carotid ultrasonography was conducive to understanding the status of ICVD,so as to determine the course and the prognosis.It was significant to clinical treatment of ICVD.

Objective To establish an in vitro renal injury model of amikacin(AKN)and investigate the protective effect and mechanism of vaccarin(VA)in the AKN-induced in vitro renal injury model.Methods Human renal tubular epithelial cells(HK-2)were cultured in vitro and incubated with different drugs of AKN or/and VA to de-termine the optimal drug concentration based on cell viability tested by MTT.The changes in intracellular oxidative stress were assessed using the dihydroethidium(DHE)probe and malondialdehyde(MDA)/glutathione(GSH)assay kits at different time points.Total RNA was extracted,and RT-qPCR was performed to detect the changes in the gene expression of kidney injury molecule-1(KIM-1)and neutropil gelatinase-associated lipocalin(NGAL).Western blot analysis was performed to detect the levels of ferroptosis-related markers solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)in HK-2 cell lysis.Results High concentrations of AKN significantly decreased the viability of HK-2 cells in vitro,with a half maximal inhibitory concentration(IC50)of(5.74±0.47)mmol/L.VA at concentrations of 25-100 μmol/L increased the viability of AKN-stimulated HK-2 cells(P<0.05).After treatment with AKN(4 mmol/L),the mRNA expression levels of KIM-1 and NGAL were significantly higher than those of the negative control(NC)group(P<0.001).VA(50 μmol/L)signifi-cantly reduced the mRNA expression levels of KIM-1(P<0.01)and NGAL(P<0.05).The intensity of DHE staining increased after 3 hours of AKN treatment,but the difference was not statistically significant.However,the intensity of DHE staining was significantly higher in the 6-24 hours group compared to the 0-hour group(P<0.01).Furthermore,MDA levels significantly increased,while GSH levels significantly decreased after 6-24 hours of AKN treatment,with statistically significant differences(P<0.05).After 6-24 hours of AKN stimula-tion,the ferroptosis-related proteins SLC7A11 and GPX4 both significantly decreased(P<0.001).Co-incubation with VA for 24 hours effectively reversed the changes in DHE staining,MDA and GSH levels,as well as the chan-ges of SLC7A11 and GPX4 protein levels(P<0.001).Conclusion In this study,an in vitro renal injury model was established by stimulating HK-2 cells with high concentrations of AKN,and it was found that VA might allevi-ate the damage to renal tubular cells caused by AKN via inhibiting oxidative stress related ferroptosis.

Objective We investigated the in vivo effects of recombinant adenovirus-associated virus type-2(AAV-2)mediated interleukin-10(IL-10)gene transfer on the expression of matrix metalloproteinase(MMP)-2,9,tissue inhibitor of metalloproteinase(TIMP)-1,collagen type Ⅰ and type Ⅲ in a rat acute myocardial infarction model.Method Male Sprague-Dawley(SD)rats were randomly divided into three groups(each n=6):sham operation group,MI/AAV2 group,and MI/AAV2-IL-10 group(1010 vg/ml×0.1 ml injection at peri-infarct regions immediately post MI).Five days later,the expressions of MMP-2 and MMP-9 were measured by RT-PCR, Western blot and zymography.The expression of TIMP-1 was measured by RT-PCR and Western blot.Collagen type Ⅰ and type Ⅲ were assessed bv RT-PCR and immunohistochemical stain.Results The myocardial expressions of MMP-2,MMP-9 and collagen contents in MI/AAV2 group were significantly increased than those in sham operation group.Myocardial expressions of MMP-2,MMP-9 were significantly decreased and the expression of TIMP-1 significantly increased in the MI/AAV2-IL-10 group than those in MI/AAV2 group. Moreover,the expressions of collagen type Ⅰ,collagen type Ⅲ and the ratio of Ⅰ/Ⅲ collagen in border zones of infarcted myocardium were decreased by 47.6%(P＜0.01),23.6%(P＜0.05),and 17.9%(P＜0.05)respectively,while the expression of TIMP-1 increased by 73.1%(P＜0.05)in MI/AAV2-IL-10 group compared to MI/AAV2 group.Conclusion In vivo myocardial IL-10 transfer reduced myocardial MMP and collagen expression and increasing the TIMP expression.

Objective Hydrocephalus is one of the most common complications of neonatal intraventricular hemorrhage (IVH). In this study, the effect of serial lumbar puncture (LP) on severe neonatal IVH is evaluated. Methods The effects of serial LP on 30 neonates hospitalized with severe IVH at Xinhua hospital since 1993 was evaluated. Another 30 neonates with severe IVH who had not been treated for preventing post-IVH hydrocephalus were used as the control group. Results Of the 30 cases which received serial LP therapy, the cerebral ventricles of 25 cases stopped enlarging and shrank distinctly. The effective rate was 83.3%. The average age when serial LP therapy initiated was (15.0±13.5) days, the average treatment course was (16.9±12.9) days. The serial LP therapy was repeated (6.4±4.7) times. The avarage interval of LP was (2.9±2.7) days and the average CSF volume removed was (6.7±1.6) ml. The average time of LP taking effect was (9.6±5.5) days. Ten cases received diamox treatment during the course of serial LP. After 1-2 weeks' combined treatment, the enlarged ventricles of 8 cases returned to normal or remained steady. Eighteen of 25 ases were followed-up for (8.3±1.9) months. Their physical and mental development were normal. Cranialcerebral ultrasound examination found that the shapes of cerebral ventricles of 14 cases were normal and the cerebral ventricles of another 4 cases enlarged slightly. The treatment with removed CSF >5 ml/time or with a shorter interval of LP (<2 d) acheived better effect (P<0.05 or 0.01). In the control group, the cerebral ventricles of 23 cases enlarged moderately or severely (6 of them complicated with hydrocephalus). Conclusions Serial LP is a safe and effective method for severe neonatal IVH in preventing post-IVH hydrocephalus.

OBJECTIVETo observe the effect of Trichinella spiralis and its worm-derived proteins on cecal ligation and puncture (CLP)-induced sepsis in mice.

METHODSEighty male BALB/c mice were randomly divided into sham-operated group, CLP group, Trichinella spiralis muscle larvae (ML) pre-infection group (ML+CLP group), soluble muscle larvae proteins (SMP) treatment group (SMP+CLP group) and excretory-secretory proteins (MES) treatment group (MES+CLP group). In ML+CLP group, the mice were orally infected with 300 Trichinella spiralis muscle larvae at 28 days before CLP and those in the other groups were intraperitioneally injected with PBS or SMP (25 µg/mice) or MES (25µg/mice) 30 min after CLP. The general condition and 72-h survival after CLP of the mice were observed. The levels of alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), creatinine (Cr), TNF-α, IL-6, IL-1β, IL-10 and TGF-β were measured at 12 h after the operation, and the pathological changes of the liver and kidney were observed.

RESULTSs Compared with the sham-operated mice, the mice in CLP group showed decreased 72-h survival, obviously increased ALT, AST, BUN, Cr, TNF-α, IL-6, IL-1β, IL-10, and TGF-β with hepatic cords disorder, hepatocytes swelling, glomerulus shrinkage, and renal tubular cell edema. Compared with CLP group, the mice in ML+CLP group showed lowered levels of ALT, AST, Cr, TNF-α and IL-1β and increased levels of IL-10 and TGF-β; in SMP+CLP group, the levels of ALT, AST, Cr, TNF-α and IL-1β were decreased and TGF-β increased. In MES+CLP group, the mice showed obviously increased 72-h survival with lowered levels of ALT, AST, BUN, Cr, TNF-α, IL-6 and IL-1β, increased levels of IL-10 and TGF-β, and alleviated liver and kidney damages.

CONCLUSIONTrichinella spiralis and its worm-derived proteins can decrease the levels of pro-inflammatory cytokines and increase immunomodulatory cytokines, and MES has more potent effect to reduce structural and functional damages of the liver and kidney.

Alanine Transaminase ; blood ; Animals ; Antigens, Helminth ; pharmacology ; Aspartate Aminotransferases ; blood ; Blood Urea Nitrogen ; Cecum ; Creatinine ; blood ; Cytokines ; blood ; Helminth Proteins ; pharmacology ; Kidney ; physiopathology ; Kidney Diseases ; therapy ; Ligation ; Liver ; physiopathology ; Liver Diseases ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Sepsis ; therapy ; Trichinella spiralis

OBJECTIVE: To clone the full-length gene encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) Chinese strain and express it in Escherichia coli. METHODS: According to the published incomplete EST (BU804141) of SjSDISP and the sequence of multiclone sites of lambda gt11 vector, 2 pairs of primers were designed and synthesized. Then the 3' and 5'ends of the EST of the SjSDISP from adult Schistosoma japonicum cDNA library were amplified by anchored PCR. After sequencing, a full-length cDNA sequence of the SjSDISP was obtained, and then it was cloned into prokaryotic expression vector pGEX-4T-1. Identified by agarosed gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for the expression under the temperature-dependent condition and Western blot analysis. RESULTS: A 1,071 bp sequence was obtained. Sequence analysis showed that the fragment contained a complete open reading frame (ORF), encoding 278 amino acid residues. This target fragment was cloned into the prokaryotic expression vector pGEX-4T-1, and expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of the expressed fusion recombinant product was 56 kD. Western blot showed that the recombinant protein was recognized by polyclonal rabbit antiserum immunized with Schistosoma japonicum adult worm antigen. CONCLUSION Cloning of the full-length gene encoding SjSDISP and its bacterial expression were successfully done.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Helminth Proteins ; biosynthesis ; genetics ; Iron-Sulfur Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; Schistosoma japonicum ; genetics ; metabolism ; Sequence Homology ; Succinate Dehydrogenase ; biosynthesis ; genetics

OBJECTIVE: This study aimed to compare the demographic characteristics and trauma service structures and processes of hospitals in 15 countries across the Asia Pacific, and to provide baseline data for the integrated trauma database: the Pan-Asian Trauma Outcomes Study (PATOS).METHODS: Medical directors and emergency physicians at PATOS-participating hospitals in countries across the Asia Pacific were surveyed through a standardized questionnaire. General information, trauma care system data, and trauma emergency department (ED) outcomes at each hospital were collected by email and analyzed using descriptive statistics.RESULTS: Survey data from 35 hospitals across 15 countries were collected from archived data between June 2014 and July 2015. Designated trauma centers were identified as the highest hospital level for trauma patients in 70% of surveyed countries. Half of the hospitals surveyed had special teams for trauma care, and almost all prepared activation protocol documents for these teams. Most hospitals offered specialized trauma education programs, and 72.7% of hospitals had a hospital-based trauma registry. The total number of trauma patients visiting the ED across 25 of the hospitals was 300,376. The overall survival-to-discharge rate was 97.2%; however, it varied greatly between 85.1% and 99.7%. The difference between survival-to-discharge rates of moderate and severe injury groups was highest in Taiwan (41.8%) and lowest in Thailand (18.6%).CONCLUSION: Trauma care systems and ED outcomes vary widely among surveyed hospitals and countries. This information is useful to build further detailed, systematic platforms for trauma surveillance and evidence-based trauma care policies.
Asia ; Asian Continental Ancestry Group ; Cross-Sectional Studies ; Education ; Electronic Mail ; Emergencies ; Emergency Service, Hospital ; Epidemiology ; Humans ; Physician Executives ; Taiwan ; Thailand ; Trauma Centers