Objective To investigate the effect of renal sympathetic denervation (RSD) on the activity of renalase in dogs with chronic heart failure (CHF).Methods After induced by abdominal aorta constriction,dogs were divided into three groups according to whether they underwent double renal artery ablation:2 dogs in control group,2 dogs in sham-operated group (no renal artery ablation),and 5 dogs in RSD group (renal artery ablation).Plasma noradrenaline (NE),B-type natriuretic peptide (BNP),and renalase were determined in 5 dogs with RSD (RSD group),2 control dogs (control group),and 2 shamoperated dogs (sham-operated group).Results NE,BNP and heart rate were significantly higher and renalase was lower in CHF group than those in control group (all P ＜ 0.05).Compared to the control dogs with CHF,the levels of renalase were significantly increased in 6 weeks after RSD [(1 948.78 ±49.19) ng/ml vs (1 847.35 ±20.72)ng/ml,P =0.029],and NE [(166.30 ±7.68)pg/ml vs (181.29 ±8.57)pg/ml],and BNP [(75.10 ± 5.58)lμg/ml vs (89.79 ± 2.04) μg/ml] were decreased in 8 weeks after RSD (all P ＜ 0.05).An decreased trend of the levels of renalase was observed in 8 weeks than in 6 weeks in CHF dogs after RSD,without significant difference (P ＞ 0.05).Conclusions The activity of renalase in dogs with CHF can be affected by RSD.

To investigate whether there is mutation in DC-SIGN promoter region in patients with chronic hepatitis B (CHB) and healthy persons previously infected with hepatitis B virus (HBV) and to explore the relationship between the mutation in dendritic cell-specific intercellular adhension molecule-3-grabbing nonintegrin (DC-SIGN) promoter region and HBV.Methods The studied population was composed of two cohorts:47 CHB patients and 20 healthy persons previously infected with HBV.The mutation in DC-SIGN promoter region was detected with PCR,single-stranded conformational polymorphism and heteroduplex analysis,cloning,sequencing and aligning the published DC-SIGN promoter sequence.Results The characteristic mutation within DCSIGN promoter region in HBV infected individuals was observed.In the DC-SIGN promoter region,4 hot spot mutations located in positions - 139,- 142,- 222,and - 336 were observed in the CHB patients,but only 1 spot mutation located in position - 139 was observed in the healthy persons previously infected with HBV.The -336C which was absent in the healthy persons previously infected with HBV was shown in 11 CHB patients (23.40％).The - 139T was far more frequent in the healthy persons previously infected with HBV ( 100％ ) than in the CHB patients (34.04％).Conclusion In the DC-SIGN promoter region,-336C may be a genetic risk factor for developing CHB,but -139T may be associated with protection against HBV.

Duck IL-18 gene was amplified from plasmid pGEM-DuIL-18 by PCR. The PCR product digested with Pst I and Xho I was inserted into eukaryotic express vector pcDNA3.1(+) to generate an recombinant expression plasmid pcDNA3.1/DuIL-18 (pDuIL-18), and transformed into Escherichia coli JM109. The recombinant colonies were identified by restriction enzyme digestion, PCR and sequencing. DNA sequence confirmed the correct sequence of the recombinant eukaryotic expression plasmid pDuIL-18 in the reading frame and the ligation part. After the transfection of pDuIL-18 into Cos7 cells, duck IL-18 mRNA was expressed in Cos7 cell. The SDS-PAGE analysis showed that the expressed duck IL-18 protein had molecular weight of 23 000 D. The results of methyl thiazolyl tetrazolium (MTT) assay showed that duck IL-18 protein expressed in Cos7 cell could induce significantly transformation of duck T lymphocytes. Immunoenhancement effect of recombinant expression plasmid pDuIL18 on avian influenza vaccine was observed by proliferation response of the T lymphocytes from spleen. It can obviously enhance the cell-mediated immune response.
Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Ducks ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunity, Cellular ; Interleukin-18 ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology ; T-Lymphocytes ; immunology ; Transfection

OBJECTIVE:To provide reference for rational use of antibiotics in the clinic. METHODS:Blood culture positive specimens were collected from our hospital during Jan. 2011-Dec.2016. Distribution of bloodstream infection(BSI)pathogens and drug resistance were analyzed in our hospital retrospectively. RESULTS:During 2011-2016,26 034 blood culture specimens isolat-ed from inpatients of our hospital were examined,including 1 775 positive specimens with positive rate of 6.82%. The specimens mainly came from tumor hematology department(10.65%),neurosurgery department(8.28%)and pediatric department(8.00%). A total of 1 775 strains of pathogens were detected,including 967 strains of Gram-negative bacteria(54.48%)mainly as Escherich-ia coli,Klebsiella pneumoniae,649 strains of Gram-positive bacteria(36.56%)mainly as Coagulase negative Staphylococci, Staphylococcus aureus and 159 strains of fungus(8.96%)mainly as Candida albicans. E. coli and K. pneumoniae were resistant to common antibiotics to different extents,but sensitive to piperacillin sodium and tazobactam sodium,imipenem,meropenem. Aci-netobacter baumanii was highly resistant to enzyme inhibitors,cephalosporins,aminoglycosides,quinolones. Pseudomonas aerugi-nosa was sensitive to third-generation cephalosporins,aminoglycosides and quinolones. S. aureus was highly resistant to penicil-lins,cephalosporins and aminoglycosides. Resistance rate of Coagulase negative Staphylococci to most commonly used antibiotics was higher than 40%. Above two bacteria were sensitive to linezolid and vancomycin with resistance rate of 0. A total of 205 strains of ESBLs-producing E. coli(42.01%),64 strains of ESBLs-producing K. pneumoniae(30.33%)and 31 strains of Methicil-lin-resistant S.aureus(17.61%)were detected.No vancomycin-resistant Enterococcus or vancomycin-resistant S.aureus was detect-ed. CONCLUSIONS:BSI pathogens mainly distribute in tumor hematology department of our hospital. BSI pathogens mainly in-clude Enterobacteriaceae and Staphylococcus,and also involve fungus. The situation of drug resistance and enzyme production are not optimistic.Antibiotics,which are sensitive to the major pathogens,include carbapenems,linezolid and vancomycin.

The highly virulent PRRSV isolate strain HN-1/06 was cultivated on Marc-145. To study the viral entry mechanisms, the GP5 gene of PRRSV isolate was amplified by RT-PCR and cloned into pcDNA3.0 to generate the expressing plasmid pcDNA-GP5. pcDNA-GP5 was transfected into 293T by the calcium phosphate precipitation method. Analysis of flow cytometry confirmed that the GP5 proteins were expressed in surface of the 293T cells. Then 293T cells were transfected with pcDNA-GP5, pHIT60 and pHIT111 plasmids to generate pseudotyping virus. The pseudotyping virus supernatant was harvested 48 hours post-transfection and was detected by Western blotting and infection assay. Western blotting indicated that the GP5 glycoproteins were incorporated into the retroviral pseudotyped virus. Infection assay showed that the pseudotyped virus infected 293T and Mark-145 cell. The pseudotyped virus could be used to further study infectious mechanism of PRRSV.
Animals ; Cell Line ; Cloning, Molecular ; Endothelial Cells ; cytology ; metabolism ; virology ; Leukemia Virus, Murine ; genetics ; metabolism ; Mice ; Porcine respiratory and reproductive syndrome virus ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transfection ; Viral Envelope Proteins ; biosynthesis ; genetics ; Virion ; genetics ; metabolism

OBJECTIVETo investigate the endovascular treatments for the ruptured aneurysms located at anterior communicating artery complex (ACoAC).

METHODSThe data of patients with ruptured ACoAC aneurysms treated in Department of Neurosurgery, First Affiliated Hospital to Fourth Military Medical University from May 2013 to December 2014 was retrospectively analyzed. Sixty-six cases were recruited including 50 male and 16 female patients. The patients aged from 31 to 69 years old, averaging (51±8) years. The Hunt-Hess grade at admission were 13 cases with grade Ⅰ, 36 cases with grade Ⅱ, 11 cases with grade Ⅲ, and 6 cases with grade Ⅳ. The most diameter of aneurysms sac: 14 cases less than or equal to 3 mm, 36 cases more than 3 mm but less than or equal to 7 mm, and 16 cases more than 7 mm. The height diameter/neck width ratio: 8 cases with absolute wide neck, 50 cases with relatively wide neck, and 8 cases with narrow neck. There were 28 cases underwent single micro-catheter embolization, 18 cases underwent double micro-catheters embolization, 14 cases underwent stent-assisted embolization and 6 cases underwent balloon-assisted embolization. The patients were followed up for 6 to 12 months and evaluated by modified Rankin score (mRS) and digital subtraction angiography (DSA). The ratio of total embolization, recurrence rate, and time from operation to reexamination of four groups managed by different endovascular treatment were compared by χ(2) test or F test.

RESULTSSixty cases were totally embolized, 3 cases subtotally embolized, 3 cases incompletely embolized. Mild hemiparalysis and aphasia occurred in 2 cases, and 1 case died of infarction induced by subarachnoid haemorrhage. The mRS at six months after operation were 0 in 31 cases, 1 in 22 cases, 2 in 8 cases, 3 in 2 cases, 4 in 2 cases, 6 in 1 case. All the included cases reexamined the DSA at averaging (7.5±1.0) month post-operatively and 4 cases recurred. There were not significant differences of the ratio of total embolization, recurrence rate, time from operation to reexamination among four groups (all P>0.05).

CONCLUSIONThe endovascular treatment maybe an ideal management for ruptured ACoAC aneurysms.

Adult ; Aged ; Aneurysm, Ruptured ; therapy ; Catheters ; Embolization, Therapeutic ; Female ; Humans ; Intracranial Aneurysm ; therapy ; Male ; Middle Aged ; Postoperative Period ; Recurrence ; Retrospective Studies ; Stents ; Treatment Outcome

Objective To investigate the clinical value of alpha-fetoprotein (AFP) combined with gamma-glutamyl transpeptidase (GGT)/aspartate aminotransferase (AST) ratio in the diagnosis of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC). Methods A total of 352 subjects who received treatment or underwent physical examination in Renmin Hospital of Wuhan University from January 15 to June 15, 2020, were enrolled, among whom there were 86 healthy controls (HC group), 68 patients with chronic hepatitis B (CHB group), 69 patients with liver cirrhosis (LC group), and 129 patients with HCC (HCC group), and a retrospective analysis was performed for the serological test results of all subjects. An analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of continuous data with skewed distribution between multiple groups, and the Nemenyi method was used for further comparison between two groups. A binary logistic regression analysis was used to calculate predictor variables; a receiver operating characteristic (ROC) curve was plotted for AFP, GGT/AST, and the predictor variables used alone or in combination, and the area under the ROC curve (AUC), sensitivity, and specificity were calculated; the Z test was used for comparison of AUC. Results The HCC group had significantly higher GGT/AST ratio and AFP than the other groups (all P < 0.05). The ROC curve analysis showed that AFP combined with GGT/AST ratio had a significantly higher AUC than AFP alone in the HCC group vs the LC group, the HCC group vs the HC+CHB+LC groups, and the HCC group vs the CHB+LC groups ( Z =2.684, 2.241, and 2.415, P =0.007, 0.025, and 0.016). Conclusion AFP combined with GGT/AST ratio can improve the clinical diagnostic performance of HBV-related HCC and thus has a certain diagnostic value.

ObjectiveTo analyze the distribution of traditional Chinese medicine （TCM） patterns as well as factors related to acute exacerbation in group E of chronic obstructive pulmonary disease （COPD）. MethodsThe general data of 161 COPD patients， including gender， age， body mass index （BMI）， disease course， smoking history， and past history， were collected. In terms of the four examinations of TCM， the differentiated patterns included phlegm-heat obstructing the lung， turbid phlegm obstructing the lung， phlegm stasis obstructing the lung， lung-spleen qi deficiency， and lung-kidney deficiency. The modified British Medical Research Council （mMRC） scale and COPD assessment test （CAT）， the pulmonary function indicators including forced expiratory volume in the first second （FEV1） and ratio of forced expiratory volume to forced vital capacity at second 1 （FEV1/FVC）， GOLD grade， and the patient's acute exacerbations in the previous year were recorded. Multivariate regression analysis was performed using logistic regression model to determine the relevant factors of patients in COPD group E. The distribution of acute exacerbations in different TCM symptom patients in group E was analyzed. ResultsThere were 80 patients （49.69%） in group E and 81 patients （50.31%） in non-group E. In group E， 23 （28.75%） patients had a history of two acute exacerbations， while 35 （43.75%） had three acute exacerbations， and 22 （27.5%） had more than three acute exacerbations. There were 13 （16.25%） cases of phlegm-heat obstructing the lung pattern， 6 （7.5%） cases of turbid phlegm obstructing the lung pattern， 8 （10%） cases of phlegm stasis obstructing the lung pattern， 22 cases （27.5%） of lung-spleen qi deficiency pattern， and 31 （38.75%） cases of lung-kidney deficiency pattern. There were significant differences in smoking history， disease course， TCM pattern， TCM syndrome score， mMRC score， and CAT score between groups （P＜0.05）. A total of 107 of the 161 patients completed pulmonary function tests， and the differences in FEV1， FEV1/FVC and GOLD grades between groups were statistically significant （P＜0.05）. Multivariate regression analysis showed that TCM pattern， TCM syndrome score and CAT score were statistically significant factors for COPD patients in group E （P＜0.05）. There were statistically significant differences in the number of acute exacerbations in different TCM patterns in group E （P＜0.05）. The patients with two acute exacerbations in the past year were mainly phlegm-heat obstructing the lung and lung-spleen qi deficiency patterns， while the three acute exacerbations were mainly seen in lung-spleen qi deficiency and lung-kidney deficiency patterns， and more than three exacerbations were more common with lung -kidney deficiency pattern. ConclusionsPatients in COPD group E were mainly the lung-spleen qi deficiency and lung-kidney deficiency patterns. Deficiency of healthy qi is the main reason for the increase in the number of acute exacerbations， and TCM patterns and CAT score were the main related factors.

OBJECTIVES: Long-term treatment of olanzapine, the most widely-prescribed second-generation antipsychotic, remarkably increases the risk of non-alcoholic fatty liver disease (NAFLD), whereas the mechanism for olanzapine-induced NAFLD remains unknown. Excessive hepatic fat accumulation is the basis for the pathogenesis of NAFLD, which results from the disturbance of TG metabolism in the liver. Apolipoprotein A5 (ApoA5) is a key regulator for TG metabolism in vivo that promotes TG accumulation in hepatocytes, thereby resulting in the development of NAFLD. However, there are no data indicating the role of apoA5 in olanzapine-induced NAFLD. Therefore, this study aims to investigate the role of apoA5 in olanzapine-induced NAFLD. METHODS: This study was carried out via animal studies, cell experiment, and ApoA5 gene knockdown experiment. Six-week-old male C57BL/6J mice were randomized into a control group, a low-dose group, and a high-dose group, which were treated by 10% DMSO, 3 mg/(kg·d) olanzapine, and 6 mg/(kg·d) olanzapine, respectively for 8 weeks. The lipid levels in plasma, liver function indexes, and expression levels of ApoA5 were detected. HepG2 cells were treated with 0.1% DMSO (control group), 25 μmol/L olanzapine (low-dose group), 50 μmol/L olanzapine (medium-dose group), and 100 μmol/L olanzapine (high-dose group) for 24 h. HepG2 cells pretreated with 100 μmol/L olanzapine were transfected with siRNA and scrambled siRNA (negative control), respectively. We observed the changes in lipid droplets within liver tissues and cells using oil red O staining and fat deposition in liver tissues using HE staining. The mRNA and protein levels of ApoA5 were determined by real-time PCR and Western blotting, respectively. RESULTS: After intervention with 3 and 6 mg/(kg·d) olanzapine for 8 weeks, there was no significant difference in body weight among the 3 groups (P>0.05). Olanzapine dose-dependently increased the plasma TG, ALT and AST levels, and reduced plasma ApoA5 levels (all P<0.05), whereas there was no significant difference in plasma cholesterol (HDL-C, LDL-C, and TC) levels among the 3 groups (all P>0.05). Olanzapine dose-dependently up-regulated ApoA5 protein levels in liver tissues (all P<0.05), but there was no significant change in ApoA5 mRNA expression among groups (P>0.05). In the control group, the structure of liver tissues was intact, the morphology of liver cells was regular, and only a few scattered lipid droplets were found in the cells. In the olanzapine-treated group, there was a large amount of lipid deposition in hepatocytes, and cells were balloon-like and filled with lipid droplet vacuoles. The nucleus located at the edge of cell, and the number of lipid droplets was increased significantly, especially in the high-dose group. Likewise, when HepG2 cells were treated with olanzapine for 24 h, the number and size of lipid droplets were significantly elevated in a dose-dependent manner. Moreover, olanzapine dose-dependently up-regulated ApoA5 protein levels in HepG2 cells (all P<0.05), but there was no significant difference in ApoA5 mRNA expression among groups (P>0.05). Compared with the HepG2 cells transfected with scrambled siRNA, the number and size of lipid droplets in HepG2 cells transfected with ApoA5 siRNA were significantly reduced. CONCLUSIONS The short-term intervention of olanzapine does not significantly increase body weight of mice, but it can directly induce hypertriglyceridemia and NAFLD in mice. Olanzapine inhibits hepatic apoA5 secretion but does not affect hepatic apoA5 synthesis, resulting in the pathogenesis of NAFLD. Inhibition of apoA5 secretion plays a key role in the development of olanzapine-related NAFLD, which may serve as an intervention target for this disease.
Animals ; Apolipoprotein A-V/genetics* ; Body Weight ; Dimethyl Sulfoxide/metabolism* ; Liver/metabolism* ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease/chemically induced* ; Olanzapine/metabolism* ; RNA, Messenger/metabolism* ; RNA, Small Interfering ; Triglycerides

Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 μL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 ³ °C/min. The volume range of 1-8 μL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 ⁶ °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
Freezing ; Vitrification ; Cryopreservation/methods* ; Trehalose ; Gold ; Rewarming ; Metal Nanoparticles ; Cryoprotective Agents ; Lasers