Background Retina fixed flat-mount perfused by Evans blue (EB) is a common method for the evaluation of blood-retinal barrier (BRB).However,previous method is inconvenient for some laboratories because the retinal specimen can not be observed by gereral microscope rather than confocal laser scanning microscope after the fixation.Objective This study was to modify the preparing way of flat-mounted retina in order to obtain transparent specimen for the observation of rat retinal vessels and the evaluation of leakage under the ordinary fluorescence microscope.Methods Forty male SD rats were divided into the control group,diabetes mellitus (DM) 1-month group,DM 3-month group and DM 6-month group according to the random number table.Streptozotocinum (STZ) of 2％ dissolved in 0.05 mmol/L sodium citrate-hydrochloric acid buffer was intraperitoneally injected in SD rats to establish DM models,and the equal volume of solvent was injected in the same way in the control rats.One month,three months and six months after injection,EB of 30 g/L was injected via rat femoral vein in the dose of 45 mg/kg.Fifteen minutes after injection of EB,the rats were sacrificed and the retinas were isolated and cut radially to prepare the flat-mounted retinas in PBS immediately and then were dried till the specimens were transparent.The specimens were examined under the fluorescence microscope.The percentage of EB leakage was quantitatively calculated by IPP 6.0 software.All procedures were performed following approval of the institutional animal care and use committee of Tianjin Medical University.Results The retina morphology was normal in the control group,and EB filled the vessels,exhibiting the red fluorescence under the fluorescence microscope.Compared with the control group,retinal background fluorescence was enhanced slightly in the DM 1-month group,and focal leakage of the EB from capillaries and focal dilated vessels were found in the DM 3-month group,further,vascular caliber inequality,retinal hypoperfusion area and a larger number of hyperfluorescence areas were seen in the DM 6-month group.The percentage of leakage area was (0.05 ±0.02) ％,(0.27 ±0.06) ％,(1.17 ±0.18)％ and (4.77 ±0.66)％ in the control group,DM 1-month group,DM 3-month group and DM 6-month group,respectively,showing a significant difference among the four groups (F =795.800,P＜0.001),and the leakage area was obviously larger in the DM 3-month group and DM 6-month group than that in thecontrol group (q'=10.338,q'=43.475,both at P＜0.001).Conclusions Modified EB-perfused retinal wholemount method is easy and helpful for clear visualization of retinal vessel leakage induced by BRB breakdown in the diabetic rats under the common fluorescence microscope.

Objective To investigate the change of plasma vitamin D, A and E levels in infant with infantile hepatitis syndrome (IHS) and to provide reliable basis for clinical treatment. Methods Seventy-three infant with IHS (IHS group) from January 2015 to May 2016 and 82 cases of physical examination infant (control group) were enrolled in this study.The levels of plasma vitamin D, A and E were tested and compared between two groups. Results The levels of vitamin D, A and E in IHS group were significantly lower than those in control group:(37.871 ± 20.111) nmol/L vs. (97.708 ± 28.827) nmol/L, (1.082 ± 0.657)μmol/L vs. (1.810 ± 0.517)μmol/L, (21.252 ± 7.596)μmol/L vs. (26.647 ± 6.495)μmol/L, P<0.01 or <0.05. The nutritional status of vitamin D, A and E in two groups had significant differences (P<0.01). Conclusions There have obvious deficiency in fat-soluble vitamin D, A and E in infant with HIS , and vitamin D deficiency is the most obvious. The levels of vitamin D, A and E are the dynamic monitoring indicators of treatment, nutritional status evaluation and long-term complications development.

Objective To study the correlations between the genetic polymorphism of brain-derived neurotrophic factor (BDNF) and the postpartum depression (PPD) in cesarean section parturient. Methods Three hundred and sixty parturients, who underwent cesarean section under spinal anesthesia from Feb. 2014 to Feb. 2015 in Third Xiangya Hospital of Central South University or Hunan Maternal and Child Health Hospital, were selected as subjects. The general information of parturients was recorded and Edinburgh Postnatal Depression Scale (EPDS) was used to evaluate the depression condition of parturients at the prenatal 1 day and the 42th day postpartum, and with a cut-off point of 12/13 for identifying PPD. The genotypes of BDNF gene locus G712A, rs56164415, rs11030100, rs11030101 and rs6265 were measured by Sequenom? Mass Array SNP. Finally, the correlations of PPD to different genotypes and general information of parturients were statistically analyzed. Results The incidence of PPD among the selected subjects was 7.2%. Pregnancy mental stress, poor pregnancy mood, perinatal elevated monocyte count, prenatal depression mood and BDNF gene locus rs6265 mutation all could affect the incidence of PPD in cesarean section parturients (P<0.05). No statistically significant difference existed between BDNF gene G712A, rs11030101, rs11030100 and rs56164415 locus mutation and PPD (P>0.05), and their haploid forms were not related to PPD also. Conclusion BDNF rs6265CC genotype, pregnancy mental stress, poor pregnancy mood, perinatal elevated monocyte count and prenatal depression mood are the risk factors for postpartum depression.

Objective To investigate the relationship between the expression of ITGAV and the radiosensitivity of NSCLC cells. Methods The expression of ITGAV in NSCLC and its relationship to the prognosis of patients who received radiotherapy were analyzed using bioinformatics methods. Differences in radiosensitivity between radio-resistant cells and parent cells were verified by clone formation experiment, and the protein expression of ITGAV was detected by Western blot. The transfection efficiency of si-ITGAV was determined by Western blot and qRT-PCR analyses. The best ITGAV interference sequence was selected to transfect A549R and H1299R cells. Clone formation experiment and flow cytometry were used to detect clone formation, apoptosis and cell cycle of A549R and H1299R cells. Results The expression of ITGAV in NSCLC tissues was significantly higher than that in normal tissues (P<0.05), and NSCLC patients with high ITGAV expression had poor prognosis. The clonogenic ability of the si-ITGAV group was significantly lower than that of the negative control group at 4, 6, 8Gy irradiation (all P<0.05). After 6 Gy irradiation, the apoptosis of the si-ITGAV group was increased (P_H1299R<0.0001, P_A549R=0.0002), the proportion of G₂/M phase cells to A549-siITGAV and H1299R-siITGAV cells was higher than that in the negative control group (P_H1299R<0.0001, P_A549R=0.0007). Conclusion Interfering with ITGAV expression can increase the radiosensitivity of NSCLC.

After participating in the student selection training and on-site refereeing of the clinical examination skills competition for college students in Sichuan-Chongqing region twice in succession, the author combined the on-site performance and competition results of the participating students with pre-match training experience and normal teaching experience. A comprehensive analysis found that the students' proficiency in basic testing skills, psychological quality and humanistic quality have an important impact on the results of the competition. Teachers should not only pay attention to the quality of basic skills training and theoretical knowledge teaching in practical teaching, but also pay attention to the cultivation of students' teamwork ability and good working habits. Therefore, the clinical laboratory skills competition has a strong leading role in promoting the education and teaching reform of medical laboratory technology and improving the quality of professional practice teaching. It is hoped that more medical colleges and universities will pay attention to and participate in it, and further promote the development of practical teaching of medical laboratory technology.

Objective To investigate the prevalence and trend of Helicobacter pylori (HP) infection in children with gastrointestinal symptoms in Kunming area, and to provide basis for prevention and cure. Methods All of 12 932 cases with gastrointestinal symptoms were adopted by retrospective analysis method from January 2013 to December 2016, and HP infection was detected by 13C-urea breath test (13C-UBT). The epidemioloqical characteristics of children with HP infection in including prevalence trend and gender and age HP were statistically analyzed. Results The rates of HP from January 2013 to December 2016 were respectively 17.85%, 18.26%, 20.41% and 32.93%.The rate of HP positive was 24.08%(3 114/12 932), which was 24.82%(1 706/6 874) in boys and 23.24%(1 408/6 058) in girls. The rate of HP infection was significantly higher in boys than that in girls, and there was statistical differences (P＜0.05). The rate of HP infection showed a gradual upward trend with age. There was statistical differences in gender between age 11 and 14[(29.03% (349/1 202) vs. 22.62% (190/840)] (P＜0.05). Conclusions HP infection rate is high in the children with gastrointestinal symptoms in Kunming, and the rate of HP infection in children is related with gender .The rate of HP infection is significantly higher in boys than that in girls, and the positive rate is directly proportional.

Objective To investigate the role of miR-125b on hepatic angiogenesis,with the hope of providing new targets for the prevention and treatment of liver fibrosis.Methods The human hepatic sinusoidal endothelial cells were transfected with miR-125b mimics and inhibitors,and the mRNA and protein expression of vascular endotheli-al growth factor(VEGF),cluster of differentiation antigens 31(CD31),von Willebrand factor(vWF),collagenⅣ,and laminin(LN)were detected by qRT-PCR and ELISA,and the expression of nitric oxide(NO)was detec-ted by fluorescent probe,scanning electron microscopy detected the alteration of the window holes on the surface of human hepatic sinusoidal endothelial cells,angiogenesis assay was performed to observe the neovascularization of each group,and dual luciferase reporter gene assay was performed to validate the targeting relationship between miR-125b and VEGF.Results qRT-PCR and ELISA showed that compared with the negative control group,the mRNA and protein expressions of VEGF,CD31,vWF,Collagen Ⅳ,and LN significantly decreased after miR-125b mimic transfection(P＜0.05),while the mRNA and protein expressions of VEGF,CD31,vWF,CollagenⅣ,and LN were significantly increased after transfection with miR-125 b mimics(P＜0.05);fluorescent probe detection showed that compared with the negative control group,the average fluorescence of intensity expression NO decreased significantly(P＜0.05),while the average fluorescence intensity expression of NO increased significant-ly after miR-125b inhibitor transfection(P＜0.05);the number of fenestrations on the surface of human liver sinu-soidal endothelial cells significantly increased after miR-125b mimic transfection(P＜0.05),while the number of fenestrations on the surface of human liver sinusoidal endothelial cells decreased significantly after miR-125 b inhibi-tor transfection(P＜0.05);angiogenesis assay showed that compared with the negative control group,the number of angiogenesis significantly decreased after miR-125b mimic transfection(P＜0.05),while the number of angio-genesis significantly increased after miR-125b inhibitor transfection(P＜0.05);dual luciferase reporter gene assay showed that compared with negative control group,the expression of relative fluorescence intensity after transfection of miR-125b mimics in VEGF wild-typ significantly decreased(P＜0.05),while the expression of relative fluores-cence intensity after transfection of miR-125b mimics in VEGF mutant significantly decreased(P＞0.05).Con-clusion miR-125b can inhibit liver angiogenesis and thus play an anti-fibrosis role,which can provide a new ref-erence for the prevention and treatment of chronic liver disease and the development of new drugs.

BACKGROUND:In recent years,the development of liver organoids has made it a hot spot in the field of international liver disease research,but there is still no article on the bibliometric analysis of liver organoids. OBJECTIVE:To explore the hot trends in liver organoids in the last 20 years based on bibliometrics and visualization analysis. METHODS:We searched the articles about liver organoids in the Web of Science Core Collection from January 1,2002 to November 12,2022.Origin,Office,and CiteSpace software were used for bibliometrics and visualization analysis.We statistically analyzed the number of annually published articles,countries,institutions,authors,journals,and keywords of the articles by generating charts. RESULTS AND CONCLUSION:The number of articles,citation frequency,institutions and personnel involved in the research about liver organoids showed an overall upward trend in the last 20 years,indicating that the field was growing rapidly and attention was increasing.The USA had published the most papers and had the strongest influence in this field.Although it had invested a lot of time and energy,the number of papers published by a single research institution in the USA was not the highest among many research institutions.China was second only to the USA in the number of publications,with the Chinese Academy of Sciences and Fudan University leading the list.Utrecht University in the Netherlands was the institution with the most publications.Clevers H was the author with the highest number of articles.The article with the highest co-citation frequency was"Long-term culture of genome-stable bipotent stem cells from adult human liver".The main fields of study for liver organoids were Molecular Science,Biology,and Immunology.The most frequently occurring keywords were stem cell,in vitro,and culture.The research hotspots in the liver organoids field were mainly focused on in vitro stem cell three-dimensional culture,differentiation and gene expression.

Objective To build the lentiviral vectors of pigment epithelial derived factor (PEDF) gene,and investigate their expression in human umbilical cord mesenchymal stem cells (hUCMSCs).Methods The PEDF lentiviral vectors (LV-PEDF) were built by DNA recombination and confirmed by DNA sequencing.hUCMSCs were transfected by LV-PEDF with MOI 10,30,50,respectively.The transfection efficiency was observed under fluorescence microscope.Cell immunofluorescence,immunocytochemistry and real-time PCR methods were used for detecting the expression of PEDF and VEGF.Results The PEDF cDNA was sub-cloned into pCDH-CMV-MCS-EF 1-copGFP vector successfully.DNA sequencing analysis confirmed that PEDF gene sequence was exactly the same with that reported in GenBank.pCDH-PEDF infected cells could show green fluorescence under fluorescence microscope.The transfection efficiency was 72.1％ in PEDF-MSCs.Immunofluorescence and immunochemical staining confirmed that PEDF protein was overexpressed in hUCMSCs.The relative expression of PEDF mRNA in experimental group and control group was (0.170±0.028) and (0.015 ± 0.007) respectively by RT-PCR,the difference was statistically significant (P＜0.00 1).The relative expression levels of VEGF mRNA in the two groups were (0.265 ± 0.022) and (0.285 ± 0.049),respectively,with no significant difference (P＞0.05).Conclusions We successfully built a lentivius vector carrying PEDF gene and obtained hUCMSCs with overexpressed PEDF.

OBJECTIVE: To investigate the association of genetic polymorphisms of norepinephrine metabolizing enzymes with postpartum depression and analyze the risk factors for postpartum depression in women following cesarean section. METHODS: A total of 591 Chinese woman of Han Nationality undergoing caesarean section were enrolled in this study. The diagnosis of postpartum depression was established for an Edinburgh Postnatal Depression Scale (EPDS) score ≥9. For all the women without antepartum depression, the genotypes of catechol-O-methyltransferase (COMT; at 5 sites including rs2020917 and rs737865) and monoamine oxidase A (rs6323) were determined using Sequenom Mass Array single nucleotide polymorphism (SNP) analysis. We analyzed the contribution of the genetic factors (SNPs, linkage disequilibrium and haplotype) to postpartum depression and performed logistic regression analysis to identify all the potential risk factors for postpartum depression and define the interactions between the genetic and environmental factors. RESULTS: The incidence of postpartum depression was 18.1% in this cohort. Univariate analysis suggested that COMT polymorphism at rs2020917 (TT genotype) and rs737865 (GG genotype) were significantly correlated with the occurrence of postpartum depression ( < 0.05). Logistic regression analysis showed that COMT polymorphism at rs2020917 (TT genotype) and rs737865 (GG genotype), severe stress during pregnancy, and domestic violence were the risk factors for postpartum depression ( < 0.05); no obvious interaction was found between the genetic polymorphisms and the environmental factors in the occurrence of postpartum depression. CONCLUSIONS The rs2020917TT and rs737865GG genotypes of COMT, stress in pregnancy, and domestic violence are the risk factors for postpartum depression.
Catechol O-Methyltransferase ; genetics ; Cesarean Section ; adverse effects ; Depression, Postpartum ; diagnosis ; enzymology ; genetics ; Domestic Violence ; psychology ; Female ; Gene-Environment Interaction ; Genotype ; Haplotypes ; Humans ; Linkage Disequilibrium ; Monoamine Oxidase ; genetics ; Norepinephrine ; metabolism ; Polymorphism, Single Nucleotide ; Postoperative Complications ; diagnosis ; enzymology ; genetics ; Pregnancy ; Pregnancy Complications ; etiology ; psychology ; Risk Factors ; Stress, Psychological