Background The scarring of conjunctival filtering blebs after glaucomatous surgery is a leading cause of operation failure.Exploring the balance between matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in conjunctival filtering bleb is very important for the study on pathogenesis of postoperative scarring.Objective This study was to evaluate the role of MMP-2 and TIMP-2 in wounding healing of subconjunctival tissue after filtering surgery in rats.Methods Sixty-three clean male SD rats were divided into normal control group and postoperative 1-day,3-day,5-day,7-day,14-day and 28-day group.The drainage tube was monocularly implanted into the anterior chamber of the rats to establish the filtering surgery models,and the operative response of the eyes was examined under the slit lamp microscope.The animals were sacrificed in corresponding time points,and the frozen sections of eyeballs were prepared,and the expressions of MMP-2 and TIMP-2 in conjunctival and subconjunctival tissues were detected using immunofluorescence technique.Western blot was employed to assay the dynamic expressions of MMP-2 and TIMP-2 proteins in conjunctival and subconjunctival tissues in different groups.The use and care of the rats complied with the Instruction Notions with Respect to Care for Laboratory by State Ministry of Science and Technology.Results Filtering blebs were formed in all the operated eyes 1 day after surgery and remained for 7-17 days,with the average survival time of 12 days.Western blot assay revealed that the expression levels of MMP-2 protein in the filtering blebs of the normal control group were 121.67 ±4.37,and the expressions were gradually elevated from the postoperative 3-day group (183.67±5.61) until the postoperative 7-day group (230.50±8.48) and then gradually declined till the postoperative 28-day group (172.33 ± 8.43),showing a significant difference among the groups (F=280.18,P＜0.05).In addition,the expression levels of TIMP-2 protein in filtering blebs of the normal control group were 102.50 ±6.25.The expression levels were gradually raised in postoperative 3-day group (162.67±7.00) and peaked in the postoperative 5-day group (232.00± 11.03),and then the levels gradually reduced till the postoperative 28-day group (150.50±6.41),with a significant difference among the groups (F =145.34,P ＜ 0.05).Immunofluorescent staining showed that MMP-2 and TIMP-2 were weakly expressed in the conjunctival epithelium of the normal rats,while in the operated rats,strong fluorescence for MMP-2 and TIMP-2 was seen in both conjunctival epithelium and subconjunctival tissues of filtering blebs.Conclusions The expression trends of MMP-2 and TIMP-2 in the filtering blebs are consistent to the fibrosis process of conjunctival tissue,indicating that MMP-2 and TIMP-2 participate in the scar formation of conjunctival filtering bleb after glaucoma filtering surgery.

Objective:To investigate the ultrasonographic characteristics and risk factors of breast cancer internal mammary lymph node (IMLN) metastasis.Methods:A retrospective analysis of 296 first diagnosed breast cancer patients in the Fourth Hospital of Hebei Medical University from March 2010 to May 2020. IMLN was divided into metastatic group (236 cases) and non-metastatic group (60 cases) based on pathology. Chi-square test and independent sample t test were used to analyze the ultrasound characteristics of IMLN metastasis and factors related to metastasis. ROC curve analysis of IMLNs were plotted to obtain the diagnostic thresholds and their sensitivity and specificity.Univariate and multivariate Logistic analysis was used to analyze the risk factors of IMLN metastasis. Results:①The appearances of IMLN in ultrasound were normal type, thickened-cortex type, unclear hilus structure type and thickened-nodular soft tissue type. ②In the two groups, the differences in IMLN long diameter, thickness diameter, number, and lymphatic hilum structure type were statistically significant (all P<0.05), and there were no significant differences in IMLN long diameter/thickness diameter and IMLN blood supply (all P>0.05). ③The long diameter threshold of IMLN for diagnosis of metastasis was 10.5 mm, the are under the ROC curve(AUC) was 0.825, with sensitivity of 58.5% and specificity 93.3%; thickness and diameter threshold was 4.5 mm, AUC was 0.790, with sensitivity 66.9% and specificity 75.0%. The sensitivity and specificity of the diagnosis of long-diameter combined structure type were 56.3% and 93.3%, respectively; the sensitivity and specificity of the diagnosis of thick-diameter combined structure type were 64.8% and 81.7%, respectively. The cortical thickness threshold was 1.9 mm, and the diagnostic sensitivity and specificity were 91.9% and 86.7%, respectively. ④The risk factors of IMLN metastasis inculded univariate analysis showed tumor length, tumor volume, axillary lymph node long diameter, axillary lymph node metastasis, and clavicle lymph node metastasis. There was a statistically significant difference in the pathology of the lower lymph nodes between the two groups ( P<0.05). Multivariate analysis showed that the long diameter of the tumor and the metastasis of the axillary lymph nodes were independent risk factors of IMLN metastasis. Conclusions:The metastatic IMLN mostly manifest as no lymphatic hilum structure or cortical thickening (≥1.9 mm), and multiple IMLN can help diagnose metastasis.Ultrasound can better assess breast cancer IMLN metastasis, and the diagnostic efficiency of IMLN long-diameter combines structure type is higher.Independent risk factors for IMLN metastasis include tumor size and axillary lymph node metastasis.

Objective To assess the diagnostic yields of clinical chromosomal microarray ( CMA) testing for patients with neurodevelopmetal disorders ( NDD) , and to characterize the spectrum of pathogenic copy number variation(CNV) in NDD.Methods The study was a cross-sectional study.NDD patients from Shanghai Children′s Medical Center ( SCMC ) from April 2014 to April 2015 were recruited.DNA samples from SCMC cohort were tested on Affymetrix Cytoscan Dx microarray platform.The diagnostic yields of CMA testing were further assessed for the whole NDD cohort and each subgroup.Results A genome-wide genotype-phenotype analysis on a total of 107 NDD cases with CMA testing was conducted.Based on the SCMC clinical cohort, the overall diagnostic yield of CMA testing for NDD patients was 20.6%(22/107). Excluding one case with chromosomal aneuploid, the frequency of non-polymorphic CNVs of the rest NDD cases were 25.5%(27/106).The diagnostic yield for developmental delay/intellectual disorder(DD/ID) and autism spectrum disorder(ASD)were 26.3% (15/57) and 10.2%(4/39) respectively.DD/ID was more likely to be associated with CNV than ASD and attention-deficit/hyperactivity disorder(ADHD).Five recurrent genomic loci were significantly enriched in patients including 1q21.1-q21.2, 15q11.2-q13.1, 22q11.2, 7q11.23 and 17q11.2.Conclusion CNV is an important pathogenesis in NDD.

Objective To study the effects of postnatal overfeeding and high-fat diet on blood pressure of rats,and to explore the pathophysiological mechanism underlying hypertension induced by continuous early postnatal overfeeding.Methods Male Sprague-Dawley rats were randomly divided into normal feeding group (10/litter) and overfeeding group (3/litter) on postnatal day 3 with a random number table.After weaning at postnatal week 3,the rats were randomly given standard chow or high-fat (HF) diet until week 16.Hence four groups were analyzed,namely normal feeding group,breastfed overfeeding group,post-weaning overfeeding group,and continuous overfeeding group.Body weight was continuously monitored in each week.Visceral fat pad (retroperitoneal and perigenital),systolic pressure,and heart rate were observed at week 3 and week 16.Thoracic aorta was sampled for measurement of vascular endothelial dilation function.Histological morphology was observed with HE staining,nitric oxide content of thoracic aorta was detected with nitrate reductase method.The mRNA expression of endothelial nitric oxide synthase (eNOS) in thoracic aorta was determined by real-time polymerase chain reaction.The protein expressions of eNOS and phosphorylated eNOS were determined by Western blot.Results At week 3,breastfed overfeeding rats displayed significantly larger body weight [(77.80 ± 0.57) g vs.(62.80 ±0.85) g,t =14.576,P ＜ 0.01] and visceral fat [retroperitoneal:(8.19 ± 0.49) mg/g vs.(4.92 ± 0.31) mg/g,t =5.629,P＜0.01;perigenital:(3.50 ±0.29) mg/g vs.(2.08 ±0.13) mg/g,t =4.552,P ＜0.01] compared with normal feedindg rats,and the protein expression of phosphorylated eNOS in aortic tissues was significantly reduced to week 16 (F =15.215,P ＜0.01);high-fat diet feeding after weaning further increased the body weight and fat mass in breastfed overfeeding rats.At week 16,continuous overfeeding rats showed hypertension [(149 ± 1.94) mmHg (1 mmHg =0.133 kPa),F =22.834,P ＜0.01],impaired vascular endothelial dilation function (F =7.648,P ＜ 0.05),and reduced protein expression of phosphorylated eNOS (F =15.215,P ＜ 0.01),while the post-weaning overfeeding group only had elevated blood pressure.Conclusions Overfeeding in breastfeeding period and high-fat diet after weaning leads to hypertension.The continuous decrease in phosphorylated eNOS in vascular tissues may be an important molecular process participating in the occurrence of vascular endothelial dysfunction in adults induced by postnatal overfeeding.

Objective To use the laboratory detection indicators and pregnant information to predict severe preeclampsia in preg-nant women.Methods 80 pregnant women with severe preeclampsia were collected as the disease group,and 444 cases were select-ed as the control group.Their laboratory detection indicators and pregnant information were compared.between the two groups.The Logistic regression and the ROC curve were used to analyze the predictive value of each indicator for predicting severe preeclamp-sia.Results BMI,MAP,HCT,PLT in the first trimester of pregnancy and BMI,MAP in the second trimester of pregnancy were positively correlated with severe preeclampsia.HCG and PAPP-A in the first trimester of pregnancy were negatively correlated with severe preeclampsia.The single indicators with large predicting value for severe preeclampsia included PAPP-A in the first trimester and MAP in the first and second trimesters of pregnancy.The combining MAP in the first and second trimester of pregnancy had the largest predictive value.Conclusion PAPP-A in the first trimester and MAP in the first and second trimester of pregnancy can predict subsequent severe preeclampsia.

AIM:The 50-Hz magnetic field (MF) is a potential health-risk factor.Its effects on the cardiovascular system have not been fully investigated .This study was conducted to explore the effects of long-term exposure to 50-Hz MF on the cardiovascular system . METHODS:In the study , an exposure system was constructed and the distribution of 50-Hz MF was detected .Sixty-four Sprague-Dawley (SD) rats were exposed to 50-Hz MF at 100 μT for 24 weeks, 20 hours per day, while another 64 rats were sham exposed. During the exposure, blood pressure was measured every 4 weeks, and 24 weeks later, echocardiography, cardiac catheterisation and electrocardiography were performed .Moreover , heart and body weight were recorded , while haematoxylin-eosin staining and real-time PCR were conducted .RESULTS:The results showed that compared with the sham group , exposure to 50-Hz MF did not exert any effect on blood pressure, pulse rate, heart rate and cardiac rhythm.Further, echocardiography and cardiac catheterisation showed that there were no significant differences in the cardiac morphology and haemodynamics .In addition , histopathological examination showed that 50-Hz MF exposure had no effect on the structure of hearts .Finally, the expression of the cardiac hypertrophic relative genes did not show any significant differences between 50-Hz MF exposure group and the sham group .CONCLUSION: Taken together , in SD rats, exposure to 50-Hz/100-μT MF for 24 weeks did not show any obvious effects on the cardiovascular system .

Objective To study the drug dosage and time dependency characteristics of high-fructose-high-fat-feeding plus dexamethason for inducing the mouse acute fatty liver model and to optimize the condition of drug induced fatty liver model.Methods Male KM mice were divided into the normal control group and high-fructose-high-fat-feeding plus peritoneal injection of dexamethason group.The mice were killed at 3 different time points.The mouse body mass and liver mass were detected.The liver index was calculated.The serum and liver tissue homogenate TG and serum glucose(GLU) levels were detected.The liver tissue pathological change was observed by HE staining.Total RNA reverse expression related gene was extracted from the liver tissue.The total protein was extracted from the liver tissue and the related protein expression was detected by Western Blot.Results Compared with the control group,blood and liver homogenate TG after 7 d in the dexamethason model group was increased,the liver index was increased,the pathological section displayed that the fatty liver was formed.RT-PCR showed that lipid metabolism related gene expression had obvious change.Western Blot showed that SIRT1 was significantly decreased.But with the dexamethason dosage decrease and time extending,the fatty liver related indexes were decreased,lipid metabolic gene PPAR,FOXO3 and FXR were gradually increased,while LXR was gradually decreased and protein SIRT1 was gradually increased.Conclusion High-fructose-high-fatfeeding plus peritoneal injection of dexamethason could establish the mouse acute fatty liver model,moreover the model maintenance has dependency on dexamethason dosage and medication time,which has a guidance significance for the drug interventional experiment.

Chemical and Drug Induced Liver Injury ; Dimethylformamide ; Humans ; Occupational Exposure

Objective To explore the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in transjugular intrahepatic portosystemic shunt (TIPS). Methods A literature search was conducted through investigating PubMed, Cochrane Library, CNKI digital library and Wanfang databases. Randomized controlled trials and cohort studies related to the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in TIPS were included in this review. According to the methods for Cochrane systematic review, the data were extracted, the quality of the literatures was evaluated, statistical analysis was conducted and the data were analyzed with RevMan 5.0. Results One randomized controlled trial and four cohort studies, including 552 patients with liver cirrhosis received TIPS, were finally included in this review. Random-effects model meta-analysis showed that puncture of the left branch of portal vein in TIPS caused lower incidence of postoperative hepatic encephalopathy than puncture of the right branch of portal vein (RR=0.51, 95%CI was 0.39 to 0.66, P<0.01). Conclusion Puncture of the left branch of portal vein in TIPS can significantly reduce the incidence of postoperative hepatic encephalopathy in comparison with puncture of the right branch of portal vein.

Objective:To verify the determination method of iodine in serum by inductively coupled plasma mass spectrometry (ICP-MS) and to evaluate the consistency between ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry in determination of serum iodine. Methods:Serum iodine concentration was determined by ICP-MS, 187Re was used as an internal standard, and ralated parameters were optimized. Eighty-eight serum samples were simultaneously determined by ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry, and the evaluation indexes included determination range of standard curve, detection limit, precision, accuracy. In addition, we also evaluated the consistency of the two methods through inter-group correlation analysis, intra-group correlation coefficient analysis, Passing-Bablok regression and Bland-Altman analysis. Results:The linear range of ICP-MS standard curve was 0 - 300 μg/L. There was a good linear correlation between iodine concentration value and iodine response value, and the correlation coefficient range was 0.999 8 to 0.999 9. The detection limit of the ICP-MS method was 1.96 μg/L. The relative standard deviation ( RSD) ranged from 0.2% to 1.4% and from 0.4% to 1.8% for intra and inter-batch precision tests of serum samples. The recovery rate ranged from 90.44% to 108.71%. The correlation analysis of 88 serum samples showed that there was a good correlation between the two methods ( r = 0.934, P < 0.05), and the intra-class correlation coefficient was 0.932. The results of Passing-Bablok regression showed that there was no significant difference between the two methods ( P > 0.05). Bland-Altman diagram suggested that the results of the two methods were consistent. Conclusions:ICP-MS method has low detection limit, high precision and accuracy. ICP-MS method is simple, rapid, easy and suitable for determination of iodine in large quantities of serum samples. The results of the two methods for determining serum iodine are consistent.