Objective To explore preliminarily the role of the E2 subunit of pymvate dehydrogenase (PDC-E2) modified by xenobiotics (e.g.2-octynic acid,2-OA) in the pathogenesis of primary biliary cirrhosis (PBC).Methods Patients of PBC (102 cases),primary sclerosing cholangitis (PSC,34 cases) and healthy controls (HC,50 cases) were selected.The anti-PDC-E2,anti-2-OA and anti-lipoic acid (LA) antibody in the peripheral blood of the 3 groups were tested by enzyme linked immunosorbent assay (ELISA).By inhibitive ELISA (iELISA),30 of the 102 PBC patients with anti-PDC-E2 antibody but without anti-2-OA antibody were selected to detect whether there was any new epitope on the PEC-E2 conjugated with 2-OA.The chi-square test and Fisher exact test were taken to analyze the enumeration data.The two-tailed unpaired t test with Welch's correction was used to compare the measurement data.Spearman rank correlation analysis was also employed for proper test.Results The positive rate of anti-PDC-E2,anti-LA and anti-2-OA antibody in PBC patients was 94.1％(96/102),73.5％(73/102) and 53.9％(55/102) respectively,all of which were statistically significantly higher than those in healthy controls group but were of no significant difference between PSC and healthy controls group.There was no significant relevance between the levels of Anti-LA and anti-2-OA antibody in the PBC group (r=-0.065,P=0.520).The iELISA results showed that the antibody,which only identified the epitopes on 2-OA-PDC-E2 induced by the 2-OA conjugation with PDC-E2,existed in 40％(12/30) of the PBC patients,and more interestingly,this antibody was predominantly appeared in PBC patients at their early clinical stage.Conclusion There are anti-LA antibody and anti-2-OA antibody in PBC patients,which have shown no significant association with each other.It is very likely that new antigenic conformational epitopes on PDC-E2 modified by 2-OA would emerge,which might led to the immune response in the individuals who are susceptible to PBC,and thus contribute for the breaking of PDC-E2 immune tolerance,and PBC occurrence finally.

Objective To study the laws of heating and thermal field of heating seed arrays in different magnetic field intensity during the magnetic induction hyperthermia.Methods 3.0 cm×3.0 cm ×2.6 cm and 5.0 cm ×5.0 cm×2.6 cm of magnetic heating seed was implanted respectively in 10.0 cm×10.0 cm ×10.0 cm pure agar phantom.The regions was divided into one file per 5 Gs in the range of 60-120 Gs magnetic induction.The temperature measurement experiments were implemented in each sub-file.Muscle-agar mixture model was used in the magnetic induction 60 Gs and 90 Gs to implement GTV-heating simulation experiments and thermal field analysis.Results The pure agar phantom heating experiments show that the heating is of basically the same rule when seeds with various diameters were implanted in the magnetic thermal array.Along with the increase of magnetic induction,the final temperature of the focal point heated for 40 min increased accordingly.and time required for heating to 50℃ was shortened correspondingly.The thermal field analysis of muscle-a mixture agar mixture model showed that the implantation density of magnetic heating seed is closely associated with the temperature of the target area and thermal field in difierent magnetic field intcnsity.Conclusions The magnetic induction has a significant impact on heating of the seeds,as the magnetic modium,and this impact is trending downwards when the magnetic field reaches a certain intensity.Within a certain size,adequate magnetic heating seed density is necessary to achieve the desired temperature required for the temperature value.The heating seeds should be put in a certain order to heat the venue evenly,not in a simply order,but in line with the principle of outer dense and inner sparse.

Objective:To analyze a Risk Scale for Emergence Agitation After General Anesthesia in Children (the EA risk scale) into simplified Chinese and evaluate the reliability and validity in children with strabismus correction surgery.Methods:After obtaining the authorization of the original author, the English version of the EA risk scale was translated, translated back and culturally debugged to form the Chinese version of the EA risk scale. Then 279 children with strabismus correction surgery were selected from a tertiary hospital of ophthalmology in Tianjin and were investigated to validate the scale.Results:The correlation coefficients of each item and the total score of the scale were respectively 0.768 (item 1) ,0.717(item 2), 0.676(item 3), 0.634(item 4) (all P < 0.01). Content validity index of the scale was 0.920, and each item was 0.80-1.00. One factors including 4 items were identified using exploratory factor analysis, accounting for 62.052% of the total variance. The optimal cut-off value for the EA risk in children was 10, with the AUG was 0.816, specificity of 0.704, and sensitivity of 0.839. The Cronbach α coefficient for the total scale was 0.819, and the intraclass correlation coefficient value between the scorers was 0.835. Conclusion:The Chinese version of the EA risk scale has good reliability and validity. The items are concise, clear, and easy to understand. It is suitable for clinical departments as a preliminary screening tool to identify emergence agitation after general anesthesia on children with strabismus correction surgery and to assess the risk of its occurrence.

Background The senescence of alveolar type II epithelial cells is an important driving factor for the progression of silicotic fibrosis, and the regulatory effects of oxamate on the senescence of alveolar type II epithelial cells is still unclear. Objective To explore whether lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cellsMethods This study was divided into two parts: in vivo experiments and in vitro experiments. In the first part, forty SPF C57BL/6J male mice were randomly divided into four groups with 10 in each group: control group, silicosis model group, low-dose oxamate treatment group, and high-dose oxamate treatment group. The silicotic mouse model was established by intratracheal instillation of 50 μL SiO₂ suspension (100 mg·mL⁻¹). The treatment models were prepared by intraperitoneal injection of 100 μL oxamate (225 mmol·L⁻¹ and 1125 mmol·L⁻¹). In the second part, induction of MLE-12 mouse alveolar type II epithelial cells was conducted with SiO₂. The in vitro experimental groups were ① SiO₂ induction groups: control group, 50 μg·mL⁻¹ SiO₂ group, 100 μg·mL⁻¹ SiO₂ group, and 200 μg·mL⁻¹ SiO₂ group, and ② oxamate treatment groups: control group, SiO₂ group (100 μg·mL⁻¹), low-dose oxamate (25 mmol·L⁻¹) treatment group, and high-dose oxamate (50 mmol·L⁻¹) treatment group. Pathological morphology of lung tissues was evaluated after hematoxylin-eosin (HE) staining; deposition of collagen in lung tissues was evaluated after sirius red staining; positive co-expression of prosurfactant protein C (Pro-SPC) and β-galactosidase was detected by immunofluorescence staining; positive expression of β-galactosidase in MLE-12 cells was detected by immunofluorescence staining. The protein expression levels of collagen type I (CoL I), fibronectin1 (FN1), hexokinase 2 (HK2), pyruvate kinase isozyme type M2 (PKM2), lactate dehydrogenase A (LDHA), p-ataxia telangiectasia and Rad3-related kinase (ATR), and cyclin-dependent kinase inhibitors p21, and p16 were detected by Western blotting. Results Compared with the control group, the protein expression levels of HK2, PKM2, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group and the SiO₂-induced MLE-12 cells (P＜0.05). The in vivo studies showed that, compared with the control group, the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the protein expression levels of CoL I, FN1, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group (P＜0.05). Compared with the silicosis model group, the oxamate treatment groups showed significant downregulation of the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the the CoL I, FN1, LDHA, p-ATR, p21, and p16 protein expression levels, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). The in vitro studies showed that, compared with the control group, the proportion of β-galactosidase positive cells and the protein expression levels of p-ATR, p21, and p16 were significantly upregulated in the SiO₂-induced group (P＜0.05). Compared with the SiO₂ group, the proportion of β-galactosidase positive cells and the LDHA, p-ATR, p21 and p16 protein expression levels were significantly downregulated in the oxamate treatment groups, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). Conclusion Lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting the senescence of alveolar type II epithelial cells.

Arg-Gly-Asp (RGD)-toxin protein Lj-RGD3 of Lampetra japonica shares homologous with a Histidine-rich glycoprotein (HRG), and both RGD-toxin protein and HRG have antiangiogenic activities with different targets. To study the relationship between the function and the structure of Lj-RGD3, we studied the anti-angiogenic characteristics of both Lj-RGD3 and the mutation named Lj-112 of which three RGD motifs of Lj-RGD3 were deleted. We synthesized the gene of Lj-112, constructed it to the plasmid pET23b, and expressed the recombinant proteins in Escherichia coli BL21. Both recombinant proteins with the C-terminal his-tag were 15 kDa soluble proteins. Then we purified rLj-RGD3 and rLj-112 using the His-Bind affinity chromatography. To examine the effect of both proteins on bFGF-induced proliferation of ECV304 cell, we carried out the 3-(4,5)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assays. For cell migration and invasion assays, we used Transwell containing insert filter and Matrigel to imitate the in vivo environment. To examine whether both proteins were capable of interrupting the angiogenesis in vivo, we used the chick chicken embryonic chorioallantoic membrane (CAM) as an angiogenesis model. We used Integrin-linked kinase1 (ILK1) ELISA method to study functionary mechanisms of rLj-RGD3 and rLj-112. Both rLj-RGD3 and rLj-112 inhibited bFGF-induced proliferation of ECV304 cells in a dose-dependent manner with IC50 at 0.889 micromol/L and 0.160 micromol/L, respectively. The results of migration and invasion assays revealed that both rLj-RGD3 and rLj-112 showed significant inhibition on bFGF induced migration and invasion of ECV304; and rLj-112 was more active than rLj-RGD3. The result of CAM angiogenesis assay demonstrated that both proteins inhibited the angiogenesis in chick CAM, and rLj-112 was more active than rLj-RGD3. ELISA assay of ILK1 showed that both rLj-RGD3 and rLj-112 down-regulated ILK1 expression of ECV304 cell. The fact of rLj-112 was more active than rLj-RGD3 on anti-angiogenesis indicate that rLj-112 was likely with histidine-rich glycoprotein (HRG), and the factor of sequence homologous between rLj-RGD3 and HRG cannot enhance antiangiogenic activities of rLj-RGD3, the signal pathway of anti-angiogenesis of rLj-RGD3 and rLj-112 are differently.
Amino Acid Sequence ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Base Sequence ; Cell Line ; Fish Venoms ; biosynthesis ; genetics ; isolation & purification ; pharmacology ; Humans ; Lampreys ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; chemistry ; pharmacology ; Oligopeptides ; biosynthesis ; genetics ; isolation & purification ; pharmacology

OBJECTIVETo investigate the effect of hyperglycemia induced by different doses of strepzotozocin (STZ) on the liver, kidneys and eyes in rats.

METHODSFifty SD rats were divided equally into 5 groups to receive intraperitoneal injections with a single dose of STZ (40, 50, or 60 mg/kg), 3 doses of 25 mg/kg STZ (given at the interval of 24 h), or no treatment (blank control). The dynamic change of blood glucose was observed within 72 h after the first injection. Blood glucose was then monitored every 3 days and the general conditions of the rats were recorded. In the 9th week, fasting blood samples were collected for biochemical analysis and the pancreas, kidney, liver, and eye were examined for pathologies.

RESULTSWithin 72 h after STZ injection, blood glucose first slightly increased and then decreased and again increased to maintain a high level. Death occurred in rats receiving injections with 50 and 60 mg/kg STZ on the third day. In the surviving rats in the 4 STZ-injected groups, the success rate of modeling was 70%, 89%, 100%, and 100%, respectively. Blood glucose showed an inverse correlation with the body weight of the rats. Cataract was observed in the 10th week in rats injected with 40 mg/kg STZ and in the 8th week in the other groups. In the 9th week, the rats receiving 40 mg/kg STZ showed normal insulin, C-peptide, urea, UA, Cr, ALT, AST, TP, and ALB levels, but the rats in the other groups all showed variations in these biochemical indices, which corresponded to the pathological findings in the pancreas, kidneys, and liver.

CONCLUSIONSThree STZ doses of 25 mg/kg is optimal and efficient for inducing diabetes in rats with stable hyperglycemia. Both fasting and random blood glucose tests contribute to the evaluation of the complications of diabetes. The eyes are the most sensitive to hyperglycemia, followed by the kidneys and then by the liver.

Animals ; Blood Glucose ; Body Weight ; Diabetes Mellitus, Experimental ; physiopathology ; Eye ; physiopathology ; Hyperglycemia ; physiopathology ; Kidney ; physiopathology ; Liver ; physiopathology ; Pancreas ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

Objective: To detect the mutation of epidermal growth factor receptor (EGFR) gene in peripheral blood of non-small cell lung cancer (NSCLC) patients in Yunnan area with Super-ARMS, and to explore its correlation with clinicopathological characteristics. Methods: A total of 222 blood samples from patients with NSCLC were collected between January 2017 to December 2018 in the Molecular Diagnostic Center of Yunnan Cancer Hospital. The EGFR gene mutation in peripheral blood samples was detected by SuperARMS, and the relationship between EGFR gene mutation and clinicopathological features was analyzed. Meanwhile, the independent risk factors influencing EFGR mutation were also analyzed. Results: In the peripheral blood of 222 NSCLC patients, there were 81 cases (36.5%) with EGFR gene mutation. Among them, exon 19 deletion and L858R gene point mutation were the most common (75.3% of total mutation); female patients had a higher mutation rate than male patients (45.9% vs 27.0%); patients <60 years old had a higher incidence of mutation than patients≥60 years old (43.2% vs 28.8%) (P<0.05 or P<0.01); moreover, patients with no history of smoking, no history of radical surgery, adenocarcinoma, advanced stage and no history of chemotherapy had higher incidence of EGFR mutation (43.9% vs 21.6%, 39.2% vs 21.2%, 43.9% vs 4.8%, 39.7% vs 23.3% and 44.0% vs 23.5%) (P<0.05 or P<0.01). Multivariate logistic analysis showed that young, no smoking history, adenocarcinoma and no surgical history were independent risk factors for EGFR gene mutation (all P<0.01). Conclusion: In the peripheral blood of patients with NSCLC in Yunnan, the mutation rate of EGFR gene is higher in patients with age<60 years old, adenocarcinoma and non-smoking. Super-ARMS method is more sensitive in the detection of EGFR mutation in peripheral blood of lung cancer patients.

Objective To evaluate the efficacy and safety of heated gel mattress(HGM) for prevention of heat loss on preterm infants with hypothermia during the transport systematically and objectively. Methods Systematic searches on PubMed, EMBASE, Cochrane Library, Web of Science CBM,CNKI,Wanfang and VIP were performed for randomized controlled trials or quasi-randomized control trials which explored the effects of HGM on prevention of hypothermia in premature infants relative to conventional alternatives. Studies were screened according to inclusion and exclusion criteria,extracting data and assessing quality. Then, Meta-analysis and trial sequential analysis were performed by using RevMan 5.3 and TSA v0.9 software developed at the Copenhagen Clinical Trials Center in Denmark, independently. Results This systematic review included 10 studies which comprised 7 randomized controlled trials and 3 quasi-randomized control trials, encompassing 773 patients. The results of Meta-analysis showed that in HGM group admission temperature on neonatal intensive care unit (standardized mean difference was 0.63, 95% confidence interval was 0.40-0.87, P=0.00), incidence of hypothermia (relative risk was 0.73, 95% confidence interval was 0.57-0.93, P=0.01) and hyperthermia (relative risk was 1.82, 95% confidence interval was 1.31-2.54, P=0.00) compared with the control group had significantly statistical difference;however, there was no significant difference in admission temperature on EM or TWM group, mortality, sepsis, retinopathy of prematurity, intraventricular hemorrhage III/IV between two groups. TSA confirmed that the pooled results of admission temperature on NICU and hyperthermia were stable and reliable; but the combination of low-temperature incidence and mortality indicators suggested that the sample size was insufficient. Conclusions HGM is an effective rewarming intervention that can improve body temperature of hypothermic preterm infants during transport, reduce the incidence of hypothermia but not decrease the incidence of morbidity and complications. However, it is recommended that clinical monitoring of body temperature should be performed dynamically to decrease the potential risk of high fever and complications. In addition, due to the limitation of quantity and quality of included studies, its cost-effectiveness and far-reaching influence on long-term follow-up outcomes need further evaluation through clinical multicenter, large sample, high-quality research.

Purpose: This study compared the diagnostic performance of the Ovarian-Adnexal Reporting and Data System (O-RADS), the Risk of Malignancy Index 4 (RMI4), the International Ovarian of Tumor Analysis Logistic Regression Model 2 (IOTA LR2), and the IOTA Simple Rules (IOTA SR) in predicting the malignancy of adnexal masses (AMs). Methods: This retrospective study included 575 women with AMs between 2017 and 2020. All clinical messages, ultrasound images, and pathological findings were collected. Two senior doctors (group I) and two junior doctors (group II) used the four systems to classify AMs. The postoperative pathological diagnosis was used as the gold standard to evaluate the diagnostic efficiency. A receiver operating characteristic curve was used to test the diagnostic performance. The interrater agreement between the two groups was tested using kappa values. Results: Of all 592 AMs, 447 (75.5%) were benign, 123 (20.8%) were malignant, and 22 (3.7%) were borderline. The intergroup consistency test yielded kappa values of 0.71, 0.92, 0.68, and 0.77 for the O-RADS, RMI4, IOTA LR2, and IOTA SR, respectively. To predict malignant lesions, the areas under the curve of the O-RADS, RMI4, IOTA LR2, and IOTA SR systems were 0.90, 0.89, 0.90, and 0.86 for group I and 0.89, 0.87, 0.88, and 0.84 for group II, respectively. The O-RADS had the highest sensitivity (91.0% in group I and 84.8% in group II). Conclusion The four diagnostic systems could compensate for junior doctors’ inexperience in predicting malignant adnexal lesions. The O-RADS performed best and showed the highest sensitivity.

Objective To investigate the risk factors of femoral head necrosis after internal fixation of fem-oral neck fracture,and to clarify the predictive role of serological index plasminogen activator inhibitors-1(PAI-1)on femoral head necrosis.Methods A total of 95 patients undergoing internal fixation for femoral neck fracture were included in the study.Relevant clinical information of patients was obtained and the serum PAI-1 levels of the patients before surgery,1,2,and 3 days after surgery were detected.After 1-year follow-up,patients were divided into necrosis group and non-necrosis group according to the occurrence of femoral head necrosis.Visual Analog Scale(VAS),Western Ontario McMaster University Osteoarthritis Index(WOMAC),and Harris Hip Score(HHS)were conducted in all patients one year after the surgery.The differences of basic clinical information and serum PAI-1 levels before and after the surgery between necrosis group and non-necrosis group were compared,and Logistic regression analysis was performed to identify the relevant risk factors for femoral head necrosis.The relationships between the PAI-1 level after operation and the VAS,WOMAC,and HHS scores of patients were figured out.The receiver operating characteristic(ROC)curve of serum PAI-1 as a predictive indicator for femoral head necrosis was drawn to clarify its predictive val-ue.Results Garden classification and reduction quality between the necrosis group and the non-necrosis group were risk factors for femoral head necrosis(P＜0.05).The serum PAI-1 at 1 day and 2 days after surgery in the necrosis group were significantly higher than that in the non-necrosis group(P＜0.05).The level of PAI-1 at 1 day and 2 days after surgery was positively correlated with VAS and WOMAC(P＜0.05),and negative-ly correlated with HHS(P＜0.05).Logistic regression analysis showed that the increase of serum PAI-1 level at 1 day and 2 days after surgery were risk factors for femoral head necrosis(P＜0.05).The ROC curve showed that serum PAI-1 level at 2 days after surgery had higher predictive value than that at 1 day after sur-gery,and the cut-off value was 44.8 ng/L,the sensitivity was 68.49％,the specificity was 86.36％,and the ar-ea under the curve(AUC)was 0.807.Conclusion The serum PAI-1 level at 1 day and 2 days after internal fixation of femoral neck fracture could be used to predict the occurrence of femoral head necrosis,especially the serum PAI-1 levels at 2 days after surgery.