Effects of interleukin-4-modified gold nanozymes on the full-thickness skin defects in diabetic mice / 中华烧伤杂志

Objective: To investigate the effects and mechanism of interleukin-4-modified gold nanoparticle (IL-4-AuNP) on the wound healing of full-thickness skin defects in diabetic mice. Methods: Experimental research methods were adopted. Gold nanoparticle (AuNP) and IL-4-AuNP were synthesized by improving the methods described in published literature. The morphology of those two particles were photographed by transmission electron microscopy, and their particle sizes were calculated. The surface potential and hydration particle size of the two particles were detected by nanoparticle potentiometer and particle size analyzer, respectively. The clearance rate of IL-4-AuNP to hydrogen peroxide and superoxide anion was measured by hydrogen peroxide and superoxide anion kits, respectively. Mouse fibroblast line 3T3 cells were used and divided into the following groups by the random number table (the same below): blank control group, hydrogen peroxide alone group treated with hydrogen peroxide only, hydrogen peroxide+IL-4-AuNP group treated with IL-4-AuNP for 0.5 h and then treated with hydrogen peroxide. After 24 h of culture, the reactive oxygen species (ROS) levels of cells were detected by immunofluorescence method; cell count kit 8 was used to detect relative cell survival rate. The macrophage Raw264.7 mouse cells were then used and divided into blank control group and IL-4-AuNP group that treated with IL-4-AuNP. After 24 h of culture, the expression of arginase 1 (Arg-1) in cells was observed by immunofluorescence method. Twelve male BALB/c mice (mouse age, sex, and strain, the same below) aged 8 to 10 weeks were divided into IL-4-AuNP group and blank control group, treated accordingly. On the 16th day of treatment, whole blood samples were collected from mice for analysis of white blood cell count (WBC), red blood cell count (RBC), hemoglobin level, or platelet count and the level of aspartate aminotransferase (AST), alanine transaminase (ALT), urea, or creatinine. The inflammation, bleeding, or necrosis in the heart, liver, spleen, lung, and kidney tissue of mice were detected by hematoxylin-eosin (HE). Another 36 mice were selected to make diabetic model, and the full-thickness skin defect wounds were made on the back of these mice. The wounds were divided into blank control group, AuNP alone group, and IL-4-AuNP group, with 12 mice in each group, and treated accordingly. On the 0 (immediately), 4th, 9th, and 15th day of treatment, the wound condition was observed and the wound area was calculated. On the 9th day of treatment, HE staining was used to detect the length of neonatal epithelium and the thickness of granulation tissue in the wound. On the 15th day of treatment, immunofluorescence method was used to detect ROS level and the number of Arg-1 positive cells in the wound tissue. The number of samples was 6 in all cases. Data were statistically analyzed with independent sample t test, corrected t test, Tukey test, or Dunnett T3 test. Results: The size of prepared AuNP and IL-4-AuNP were uniform. The particle size, surface potential, and hydration particle size of AuNP and IL-4-AuNP were (13.0±2.1) and (13.9±2.5) nm, (-45.8±3.2) and (-20.3±2.2) mV, (14±3) and (16±4) nm, respectively. For IL-4-AuNP, the clearance rate to hydrogen peroxide and superoxide anion were (69±4)% and (52±5)%, respectively. After 24 h of culture, the ROS level of 3T3 in hydrogen peroxide alone group was significantly higher than that in blank control group (q=26.12, P<0.05); the ROS level of hydrogen peroxide+IL-4-AuNP group was significantly lower than that in hydrogen peroxide alone group (q=25.12, P<0.05) and close to that in blank control group (P>0.05). After 24 h of culture, the relative survival rate of 3T3 cells in hydrogen peroxide+IL-4-AuNP group was significantly higher than that in hydrogen peroxide alone group (t=51.44, P<0.05). After 24 h of culture, Arg-1 expression of Raw264.7 cells in IL-4-AuNP group was significantly higher than that in blank control group (t'=8.83, P<0.05).On the 16th day of treatment, there were no significant statistically differences in WBC, RBC, hemoglobin level, or platelet count and the level of AST, ALT, urea, or creatinine of mice between blank control group and IL-4-AuNP group (P>0.05). No obvious inflammation, bleeding or necrosis was observed in the heart, liver, spleen, lung, and kidney of important organs in IL-4-AuNP group, and no significant changes were observed compared with blank control group. On the 0 and 4th day of treatment, the wound area of diabetic mice in blank control group, AuNP alone group, and IL-4-AuNP group had no significant difference (P>0.05). On the 9th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 9.45 and 14.87, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=5.42, P<0.05). On the 15th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 4.84 and 20.64, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=15.80, P<0.05); moreover, inflammations such as redness and swelling were significantly reduced in IL-4-AuNP group compared with the other two groups. On the 9th day of treatment, compared with blank control group and AuNP alone group, the length of neonatal epithelium in the wound of diabetic mice in IL-4-AuNP group was significantly longer (all P<0.05), and the thickness of the granulation tissue in the wound was significantly increased (with q values of 11.33 and 9.65, respectively, all P<0.05). On the 15th day of treatment, compared with blank control group, ROS levels in wound tissue of diabetic mice in AuNP alone group and IL-4-AuNP group were significantly decreased (P<0.05). On the 15th day of treatment, the number of Arg-1 positive cells in the wounds of diabetic mice in IL-4-AuNP group was significantly more than that in blank control group and AuNP alone group, respectively (all P<0.05). Conclusions: IL-4-AuNP is safe in vivo, and can improve the oxidative microenvironment by removing ROS and induce macrophage polarization towards M2 phenotype, thus promote efficient diabetic wound healing and regeneration of full-thickness skin defects in diabetic mice.

Clinical value of plasma miR-27 expression in patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the clinical value of microRNA(miRNA, miR)-27 expression in patients with diabetic retinopathy(DR).<p>METHODS: A total of DR 80 patients(DR group)treated between January 2019 and January 2020 were retrospectively reviewed. Meanwhile, 40 patients with simple type 2 diabetes mellitus(T2DM)(T2DM group)and 40 normal healthy persons(control group)were enrolled, and plasma RNA was extracted. Real time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)was adopted to determine plasma miR-27 expression, and enzyme-linked immunosorbent assay was performed to determine the vascular endothelial growth factor(VEGF)level. Plasma miR-27 and serum VEGF expression in different groups and in patients with different severities of DR was comparatively analyzed. Multivariate Logistic regression analysis was performed to screen factors influencing the expression of miR-27 in patients with DR, and Pearson correlation analysis of miR-27, serum VEGF and blood glucose indexes was conducted. Meanwhile, significance of miR-27 in pathogenesis of DR was summarized.<p>RESULTS: DR group had the highest plasma miR-27 and serum VEGF levels, followed by T2DM group, and then the control group(<i>P</i><0.05). Proliferative diabetic retinopathy(PDR)patients had higher levels of plasma miR-27, serum VEGF, fasting blood glucose and glycated hemoglobin than those with non-proliferative diabetic retinopathy(NPDR)(<i>P</i><0.05). It was found that course of disease(<i>OR</i>=3.206), fasting blood glucose(<i>OR</i>=2.570), glycated hemoglobin(<i>OR</i>=2.787), VEGF(<i>OR</i>=3.442)and severity of DR(<i>OR</i>=5.842)were influencing factors of plasma miR-27 expression in DR patients(<i>P</i><0.05). In DR patients, relative expression of plasma miR-27 was positively correlated with serum VEGF, fasting blood glucose and glycated hemoglobin(<i>r</i>=0.548, 0.398, 0.522, all <i>P</i><0.05).<p>CONCLUSION: DR patients have higher plasma miR-27 expression level than those with simple T2DM and normal healthy people. The duration of diabetes, fasting blood glucose, glycated hemoglobin and severity of DR all affect the expression of miR-27. Besides, miR-27 is positively correlated with serum VEGF, glycated hemoglobin and fasting blood glucose. It is speculated that miR-27 may mediate the pathogenesis and progression of DR by regulating glucose metabolism and promoting angiogenesis.

Research Progress on Forensic Palynology and Its Application in Forensic Science / 法医学杂志

Forensic palynology is to apply palynology to the field of forensic science, using pollen and spores to solve issues in juridical practice, such as civil and criminal issues. Sporopollens have a small size, wide distribution, diverse morphology, can be easily transferred, have durability, and is not easily noticed. It can provide strong investigation and related evidence for case detection as good trace evidence. The application of palynology in forensic science has achieved certain success, but it is underutilized in most countries. This paper analyzes the evidence value provided by sporopollen, collection of the sporopollen samples, the progress in detection technology and challenges ahead, based on the biological characteristics of sporopollen, combined with recent successful cases in forensic science, to comprehensively discuss the research progress in forensic palynology and its application prospects in forensic science.

Contribution of Quick Sequential Organ Failure Assessment Score Combined with Electrocardiography in Risk Stratification of Patients with Acute Pulmonary Embolism / 中华医学杂志(英文版)

<p><b>Background</b>The quick Sequential Organ Failure Assessment (qSOFA) score emerged recently. We investigated its contribution to risk stratification in acute pulmonary embolism (PE) by combining with electrocardiography (ECG).</p><p><b>Methods</b>Acute PE patients diagnosed in Beijing Chao-Yang Hospital, Capital Medical University, from 2008 to 2018 were retrospectively studied and divided into high- and low - risk groups by imaging and biomarkers. The ECG scores consisted of tachycardia, McGinn-White sign (SQT), right bundle branch block, and T-wave inversion of leads V-V. A new combination of qSOFA scores and ECG scores by logistic regression for predicting high-risk stratification patients with acute PE was evaluated by a receiver operating characteristic curve.</p><p><b>Results</b>Totally 1318 patients were enrolled, including 271 in the high-risk group and 1047 in the low-risk group. A combination predictive scoring system named qSOFA-ECG = qSOFA score + ECG score was created. The optimal cutoff value for qSOFA-ECG was 2, and the sensitivity, specificity, positive predictive value, and negative predictive value were 81.5%, 72.3%, 43.2%, and 93.8%, respectively. For predicting high-risk stratification and reperfusion therapy, the qSOFA-ECG is superior to PE Severity Index (PESI) and simplified PESI.</p><p><b>Conclusions</b>The qSOFA score contributes to identify acute PE patients with potentially hemodynamic decompensation that need monitoring and possible reperfusion therapy at the emergency department arrival when used in combination with ECG score.</p>

Clinical Characteristics and Current Interventions in Shock Patients in Chinese Emergency Departments: A Multicenter Prospective Cohort Study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Shock is a life-threatening condition in emergency departments (EDs) and is associated with a high mortality; however, its clinical characteristics and current interventions in China are seldom reported. This study investigated the clinical characteristics and current interventions of shock patients in Chinese EDs.</p><p><b>METHODS</b>This multicenter prospective cohort study was conducted in the EDs of 33 academic hospitals in 16 Chinese provinces. Adult shock patients were enrolled from December 2013 to April 2014. Age, sex, comorbidities, shock subtype, and vital signs were recorded on ED arrival; details of subsequent interventions and treatments were added. We compared those data between survivors and nonsurvivors. All patients were followed up for 3 days. The primary outcome was 3-day mortality. Binary logistic regression analysis identified the independent predictors of that mortality.</p><p><b>RESULTS</b>We enrolled 1095 shock patients. The 3-day mortality was 27.5%, 36.3%, and 29.0%, respectively, in the whole cohort and for cardiogenic and septic shock. Within the first 24 h, 1039 patients (94.9%) were admitted to the Intensive Care Unit. Use of bicarbonate, epinephrine, and dopamine is an independent predictor for mortality. Hemorrhage and trauma (39.1%), along with sepsis (40.4%) were the most commonly observed causes of shock in the ED. In nondiabetic patients with cardiogenic shock, 3-day mortality was 69.2% for patients needing glucose control - much higher than in those not needing glucose control (30.0%, P= 0.01). Hydroxyethyl starch (HES) was applied in 29.6% of septic shock patients, and the mortality of septic patients who received HES was much higher than those who did not (38.2% vs. 25.1%, P = 0.006).</p><p><b>CONCLUSIONS</b>In China, short-term mortality of shock patients in EDs is still high, especially among those with cardiogenic and septic shock. HES application needs to be restricted - particularly in septic shock patients.</p>

Clinical analysis of 9 cases of anti-NMDAR encephalitis / 解放军医学杂志

Objective: To explore the pathogenesis and clinical features of anti- N-methyl-D-aspartate receptor (NMDAR) encephalitis. Methods: The clinical characteristics of 9 patients with anti-NMDAR encephalitis in Department of Neurology, Army General Hospital of Chinese PLA from October 2013 to October 2015, were retrospectively analyzed. And by literatures review, the pathogenesis, clinical feature, diagnosis, treatments and prognosis of this disease were summarized. Results: The average age of the 9 patients (3 men and 6 women) was 39 years, 5 of them had precursor nonspecific infection. The initial symptom of neurologic system was mental-behavior disorder in all the 6 females, or epilepsy in all the 3 males. Mental-behavior disorder existed in all the 9 cases during the disease course, including 7 cases of epilepsy, 8 cases of consciousness disturbance, 5 cases of involuntary movement, and 3 cases of inadequate ventilation. The anti-NMDAR antibodies were positively detected from the cerebrospinal fluid and blood of all the 9 cases. After gamma globulin and hormone therapy, 8 cases were cured or better, and only 1 case invalid. A six months follow-up found that 8 cases were independent with eusemia and 1 case got disability. Conclusions: The clinical characteristics of anti-NMDAR encephalitis are mental-behavior disorder, or epilepsy upon the basis of a precursor infection, with positive anti-NMDAR antibodies detected from the cerebrospinal fluid and blood. Gamma globulin and hormone therapy may help the clinical recovery.

Puerarin protects human bronchial epithelial cells from apoptosis induced by gunpowder smog / 解放军医学杂志

Objective To investigate protective effects of puerarin on the human bronchial epithelial (BEAS-2B) cell line against apoptosis caused by gunpowder smog and its mechanisms. Methods BEAS-2B cells cultured in vitro were randomly divided into control group, smog group (the group treated with 4g gunpowder smog for lOmin), and smog + puerarin group [puerarin groupj the cells were pre-incubated with various concentrations of puerarin (12.5, 25.0, 50.0, 100.0µg/ml) and then exposed to smoke]. Puerarin was added into the cells after innoculation for 12h and then the cells were sequentially cultured for 24h and followed by exposure to smoke for lOmin. After being cultured again for 2h, the smoked cells were examined for cell viability using Cell Counting Kit-8(CCK-8); cell apoptosis was observed using Hoechst33258 nucleus staining, and positive rates of Annexin V-PI staining cells and caspase-3 were determined with flow cytometer. Results Compared with control, treatment of BEAS-2B cells with 4g gunpowder smog induced a characteristic ap op to tic cell death (P<0.01). Pretreatment with various concentrations of puerarin antagonized the action of gunpowder smog in different degrees. The 25µg/ml was determined as the optimal effective concentration of puerarin. Compared with smog group, the apoptosis rate of BEAS-2B cells and positive rates of Annexin V-PI staining cells and caspase-3 were decreased significantly in smog + puerarin group (P<0.05, P<0.01). Conclusion Gunpowder smog can induce apoptosis of BEAS-2B cells in vitro, while pretreatment with puerarin could protect BEAS-2B cells against apoptosis induced by gunpowder smog.

Establishment of reverse genetics system for class I NDV08-004 strain / 病毒学报

Based on the genomic sequence of NDV08-004 strain (GenBank accession number FJ794269), seven pairs of primers were designed to amplify the genomic fragments by RT-PCR and cloned into pGEM-Teasy vector. The fragments (named A to G) were sub-cloned into transcription vector pOLTV5 according to the universal RE site and the plasmid named NDV08-004-pO which contained the full length cDNA of NDV08-004 strain was constructed. Three helper plasmids (pCI-NP, pCI-P and pCI-L) together with NDV08-004-pO were co-transfected into BSR T7/5 cells, and the transfection supernatant was inoculated into SPF embryonated eggs to rescue the virus. The virus was rescued successfully and identified by HA and RT-PCR and sequencing. The rescue system constructed in this study provided a good foundation for the further related research.

Risk factors on the recurrence of ischemic stroke and the establishment of a Cox' s regression model / 中华流行病学杂志

Objective To investigate the risk factors and establish the Cox' s regression model on the recurrence of ischemic stroke. Methods We retrospectively reviewed consecutive patients with ischemic stroke admitted to the Neurology Department of the Hebei United University Affiliated Hospital between January 1,2008 and December 31,2009. Cases had been followed since the onset of ischemic stroke. The follow-up program was finished in June 30, 2010. Kaplan-Meier methods were used to describe the recurrence rate. Monovariant and multivariate Cox' s proportional hazard regression model were used to analyze the risk factors associated to the episodes of recurrence.And then, a recurrence model was set up. Results During the period of follow-up program, 79 cases were relapsed,with the recurrence rates as 12.75% in one year and 18.87% in two years. Monovariant and multivariate Cox' s proportional hazard regression model showed that the independent risk factors that were associated with the recurrence appeared to be age (X1)(RR=1.025,95% CI: 1.003-1.048),history of hypertension (X2) (RR= 1.976, 95% CI: 1.014-3.851), history of family strokes (X3) (RR=2.647,95%CI: 1.175-5.961), total cholesterol amount (X4) (RR= 1.485,95%CI: 1.214-1.817), ESRS total scores (X5) (RR= 1.327,95%CI: 1.057-1.666) and progression of the disease (X6) (RR= 1.889,95%CI: 1.123-3.178). Personal prognosis index (PI) of the recurrence model was as follows: PI=0.025X1 + 0.681X2+ 0.973X3 + 0.395X4+ 0.283X5 + 0.636X6. The smaller the personal prognosis index was, the lower the recurrence risk appeared, while the bigger the personal prognosis index was, the higher the recurrence risk appeared. Conclusion Age, history of hypertension, total cholesterol amount, total scores of ESRS, together with the disease progression were the independent risk factors associated with the recurrence episodes of ischemic stroke. Both recurrence model and the personal prognosis index equation were successful constructed.

The Substrate Specificity of Cyclic Imide Hydrolase Mutants / 中国生物工程杂志

The effect of C-terminal region residues on the substrate specificity of a novel cyclic imide hydrolase (CIH), a recombinant cyclic imide hydrolase (CIH293), and its mutants deleted or substituted at C-terminus (CIH291, CIH290, KK292-293EE) was reported. The substrate specificity and kinetic parameters of the mutants were analyzed by both the spectrophotometric assay and high-performance liquid chromatography. Results show that the substrate specificity of mutants was not obviously changed, but slightly low for the affinity between the substrate and enzyme, compared with the wild-type enzyme, CIH293. In conclusion, the last three residues of CIH293 play an important role for the enzyme activity.