[Treatment efficacy of dietary supplement Licofor for dry eye associated with meibomian gland dysfunction].

Objective: To evaluate the clinical efficacy of dietary supplement Licofor in the treatment of dry eye associated with meibomian gland dysfunction (MGD). Methods: This was a prospective, randomized controlled clinical trial. Sixty patients [25 males, 35 females, aged (42±13) years] who had dry eye associated with MGD were recruited in Xiangya Hospital of Central South University from December 2018 to October 2019. The patients were equally divided into two groups: 30 cases (60 eyes) in the experimental group and 30 cases (60 eyes) in the control group. All subjects were treated with eye hot compress, artificial tears and antibiotic ointment. After that, the experimental group and control group were received dietary supplementary Licofor or placebo daily for 12 weeks. The symptoms and signs of dry eye, morphology and function of meibomian gland, and inflammatory response were assessed at the beginning, 4th, 8th and 12th week of treatment. Results: After 12 weeks of treatment, statistically significant improvements in ocular surface disease index (OSDI) scores, tear break-up time (TBUT), corneal fluorescein staining (CFS), the morphology of eyelid margin, meibomian gland orifice, meibomian gland expressibility, meibum quality, and periglandular inflammatory cell density were determined in both groups (all P<0.05). In the Licofor group, the improvement of OSDI scores [16.7 (12.5, 20.8) vs 20.8 (18.8, 22.9), P<0.001], the morphology of eyelid margin, meibomian gland orifice and periglandular inflammatory cell density [443 (318, 513) vs 553 (415, 676)/mm2, P=0.002] were more significant (all P<0.05). Conclusion: The combined treatment of licofor and conventional treatment can significantly improve symptoms of dry eye, the morphology of eyelid margin, meibomian gland orifice, meibum quality, and eyelid inflammation response of dry eye associated with MGD.

[Clinical analysis of peri-operative gastrointestinal complications after endovascular and open repair of abdominal aortic aneurysm].

Objective: To investigate the etiology, treatment method and prevention of gastrointestinal complications(GCs) after endovascular and open repair of abdominal aortic aneurysm (AAA). Methods: The clinical data of 716 cases who were diagnosed as AAA and underwent endovascular(EVAR) or open repair (OR) from Department of Vascular Surgery, Beijing Anzhen Hospital, Capital Medical University April 2009 to March 2017 were collected and analyzed retrospectively. There were 608 males (84.9%)and 108 females(15.1%), aging of 69.4 years (range: 52-86 years). There were 539 cases(75.3%) underwent EVAR and 177 cases(24.7%) underwent OR. The morbidity of GCs and mortality of GCs, such as acute pancreatitis, cholecystitis, ischemic colitis, intestinal obstruction and peptic ulcer, between EVAR and OR group were compared. The treatment of the GCs and the prognosis of the patients were reported. Results: The morbidity of GCs in EVAR and OR group were 4.6%(25/539)and 35.0%(62/177), respectively. There were 10 cases and 28 cases suffering from acute pancreatitis in EVAR and OR group, respectively; 4 cases and 6 cases suffering from cholecystitis in the two groups; 6 cases and 13 cases suffering from ischemic colitis in the two groups; 5 cases suffering from intestinal obstruction in OR group; 5 cases and 10 cases suffering from peptic ulcer in the two groups. Two patients died in EVAR group, and the peri-operative mortality was 0.37%, one died of ischemic colitis with acute myocardial infarction, the other died of ischemic colitis with septic shock. Six patients died in OR group, and the peri-operative mortality was 3.39%, two patients died of acute pancreatitis with intestinal necrosis, one patient died of cholangitis with peritonitis and septic shock, three patients died of ischemic colitis with acute renal failure or septicemia. Conclusions: The etiology of peri-operative GCs after AAA repair may include inferior mesenteric artery occlusion or ligation, pancreas injury, organ hypoperfusion and so on.

[Analysis of risk factors for cerebrovascular complications after carotid endarterectomy].

Objective: To evaluate the related risk factors of cerebrovascular complications after carotid endarterectomy (CEA) and to improve the efficacy of CEA in the treatment of ischemic stroke. Methods: The clinical data of 295 patients with atherosclerotic carotid artery stenosis who underwent CEA in the Department of Vascular Surgery, Beijing Anzhen Hospital, Capital Medical University from January 2013 to March 2017 were retrospectively analyzed. Results: As the results of the single-factor analysis of logistics, severe lower limb artery stenosis (RR=5.667, P=0.017), systolic blood pressure before the carotid artery clamping (RR=6.659, P=0.010), diastolic blood pressure before the carotid artery clamping (RR=3.981, P=0.046), stump pressure (RR=5.359, P=0.021), diastolic blood pressure after surgery (RR=9.550, P=0.002), diastolic blood pressure of the first day after surgery (RR=7.932, P=0.005) were influencing factors of postoperative cerebrovascular complications after CEA. The results of multi-factor analysis of logistic regression indicated that diastolic blood pressure before the carotid artery clamping (RR=0.953, P=0.024) and stump pressure to basic systolic blood pressure index (SSI)>0.25 (RR=0.086, P=0.049) were independent risk factors for postoperative cerebrovascular complications after CEA. Conclusion: Systolic blood pressure before carotid artery clamping and SSI>0.25 are independent risk factors for postoperative cerebrovascular complications after CEA. Close follow-up and drug treatment for patients after CEA might be beneficial to reduce postoperative carotid artery restenosis.

Establishment and application of molecular ID in the main inbred lines of Chinese cabbage.

Chinese cabbage is an important cruciferous vegetable in China. The differences in the morphology and other characteristics of the different varieties of Chinese cabbage are generally caused by their different genes. Using the simple sequence repeat (SSR) DNA molecular markers is an effective way to identify different genotypes. The identification of a genetic relationship is a key point in the breeding process, and it plays an important role in guiding parent selection and breeding of high-yield varieties. Moreover, the establishment of genomic fingerprints is significant for plant variety protection. Three to five SSR sites were selected from each of the 10 Chinese cabbage chromosomes on the basis of the abundance of SSR loci on them. According to the differences in the SSR polymorphic bands, a genomic fingerprint comprising 36 different loci was established in the 20 main inbred lines of Chinese cabbage, and this fingerprint was converted digitally into a molecular ID with 36 numbers based on the 36 SSR sites. The utility of this core set SSRs was demonstrated in 20 main inbred lines of Chinese cabbage, which could be placed into six clusters that were largely consistent with previous classification based on morphology data. Moreover, the molecular ID of an F1 hybrid can be deduced from its parents molecular IDs, and its purity can be determined by selecting one or two SSR loci from the 36 different loci.

Proteomic and gene expression analyses during bolting-related leaf color change in Brassica rapa.

Bolting and flowering are key processes during the growth and development of Chinese cabbage (Brassica rapa L. ssp pekinensis). Understanding the molecular mechanisms underlying bolting and flowering is of significance for improving production of the vegetable. A leaf-color change from bright green to gray-green has been observed following differentiation of the flowering stem and before bolting in the vegetable, and is considered to be a signal for bolting. Proteomics in meristem tissues of an inbred line (C30) were analyzed by two-dimensional electrophoresis during the transition period. We found that some proteins were specifically expressed while others were differentially expressed. Among these, 17 proteins were specifically expressed before the color change, 18 were specifically expressed after the color change, 21 were downregulated during the color change, and 29 were upregulated. Mass spectrometric analysis (MALDI-TOF-TOF/MS) was used to analyze 17 protein spots, and four proteins (subunit E1 of vacuolar-type H+ transporter ATPase, the large subunit of Rubicon, S-adenosylmethionine synthetase, and tubulin α-2) were identified. qPCR analysis was conducted to quantify the expression of genes encoding these proteins during the transitional period. The expression of BrVHA-E1, BrSAMS, BrrbcL, and BrTUA6 was significantly different before and after the leaf-color change, suggesting that these genes might be involved in regulating flower differentiation and bolting.

Serum soluble CD40 ligand levels after acute intracerebral hemorrhage.

BACKGROUND: Soluble CD40 ligand (sCD40L) is associated with inflammation. This study aimed to assess the prognostic value of sCD40L for clinical outcomes of acute intracerebral hemorrhage (ICH) patients. MATERIALS AND METHODS: The serum sCD40L levels of 110 patients and 110 age- and gender-matched healthy controls were measured using sandwich immunoassays. The relationships between serum sCD40L levels and 1-week mortality, 6-month mortality, 6-month overall survival, 6-month unfavorable outcome (modified Rankin Scale score >2), and ICH severity including hematoma volume and National Institutes of Health Stroke Scale (NIHSS) score were assessed using multivariate analysis. RESULTS: Compared with healthy controls, ICH patients had higher serum sCD40L levels. Serum sCD40L levels were correlated positively with hematoma volumes and NIHSS scores using a multivariate linear regression. Multivariate analysis results indicated that sCD40L was identified an independent predictor of 1-week mortality, 6-month mortality, 6-month unfavorable outcome and 6-month overall survival. sCD40L also showed high predictive performances for 1-week mortality, 6-month mortality and 6-month unfavorable outcome based on receiver operating characteristic curve. CONCLUSIONS: Elevated serum sCD40L levels are independently associated with ICH severity and clinical outcomes. And sCD40L has potential to be a good prognostic biomarker of ICH.

The expression of human alpha -like globin genes in transgenic mice mediated by bacterial artificial chromosome.

After screening a bacterial artificial chromosome of human genomic DNA library with human HS-40, zeta-, alpha-, and theta-globin probes, a 110-kb clone bearing the whole human alpha-globin gene cluster was obtained and rare restriction endonuclease mapping was performed. The bacterial artificial chromosome DNA was isolated, and transgenic mice were generated. Three founders were detected from 35 newborn mice. The copy numbers were 1, 2, and 2, and the expression of human alpha-globin genes in various tissues at different developmental stages in the transgenic mice was assayed. The human alpha-globin mRNA can be detected in bone marrow, kidney, liver, brain, but not in muscle, testis, or thymus. The human zeta-globin genes were switched off, and the alpha-globin genes were switched at day 11.5 in mouse embryo, indicating that developmental stage-specific expression of the alpha-like globin genes was properly regulated. The human alpha-globin mRNA ranged between 17-68% of the endogenous mouse alpha-globin, suggesting that the expression of human alpha-globin genes is integration site-dependent in transgenic mice. The ratio of human alpha(2)- and alpha(1)-globin gene expression in adult transgenic mouse is about 2.5:1 similar to the expression in human.

Regulation of the transcription factor GATA-1 at the gene and protein level.

GATA-1, an important hematopoietic transcription factor, plays a critical role in differentiation and maturation of erythroid and megakaryocytic cell lines. GATA-1 appears to serve as a factor for virtually all characterized erythroid and megakaryocytic-expressed genes. Thus, defining the mechanisms by which the GATA-1 gene and protein are regulated should provide important clues regarding the establishment of erythroid and megakaryocytic programs of gene expression in committed cells and their maintenance thereafter in maturing precursors. This review focuses on the regulation of GATA-1 expression and elucidates the regulation of GATA-1 at the gene and protein levels. Such research is expected to provide insights into the mechanisms involved in hematopoietic commitment.

Trapping of inhibitor-induced conformational changes in the erythrocyte membrane anion exchanger AE1.

AE1, the chloride/bicarbonate anion exchanger of the erythrocyte plasma membrane, is highly sensitive to inhibition by stilbene disulfonate compounds such as DIDS (4,4'-diisothiocyanostilbene-2, 2'-disulfonate) and DNDS (4,4'-dinitrostilbene-2,2'-disulfonate). Stilbene disulfonates recruit the anion binding site to an outward-facing conformation. We sought to identify the regions of AE1 that undergo conformational changes upon noncovalent binding of DNDS. Since conformational changes induced by stilbene disulfonate binding cause anion transport inhibition, identification of the DNDS binding regions may localize the substrate binding region of the protein. Cysteine residues were introduced into 27 sites in the extracellular loop regions of an otherwise cysteineless form of AE1, called AE1C(-). The ability to label these residues with biotin maleimide [3-(N-maleimidylpropionyl)biocytin] was then measured in the absence and presence of DNDS. DNDS reduced the ability to label residues in the regions around G565, S643-M663, and S731-S742. We interpret these regions either as (i) part of the DNDS binding site or (ii) distal to the binding site but undergoing a conformational change that sequesters the region from accessibility to biotin maleimide. DNDS alters the conformation of residues outside the plane of the bilayer since the S643-M663 region was previously shown to be extramembranous. Upon binding DNDS, AE1 undergoes conformational changes that can be detected in extracellular loops at least 20 residues away from the hydrophobic core of the lipid bilayer. We conclude that the TM7-10 region of AE1 is central to the stilbene disulfonate and substrate binding region of AE1.

Topology of the membrane domain of human erythrocyte anion exchange protein, AE1.

Anion exchanger 1 (AE1) is the chloride/bicarbonate exchange protein of the erythrocyte membrane. By using a combination of introduced cysteine mutants and sulfhydryl-specific chemistry, we have mapped the topology of the human AE1 membrane domain. Twenty-seven single cysteines were introduced throughout the Leu708-Val911 region of human AE1, and these mutants were expressed by transient transfection of human embryonic kidney cells. On the basis of cysteine accessibility to membrane-permeant biotin maleimide and to membrane-impermeant lucifer yellow iodoacetamide, we have proposed a model for the topology of AE1 membrane domain. In this model, AE1 is composed of 13 typical transmembrane segments, and the Asp807-His834 region is membrane-embedded but does not have the usual alpha-helical conformation. To identify amino acids that are important for anion transport, we analyzed the anion exchange activity for all introduced cysteine mutants, using a whole cell fluorescence assay. We found that mutants G714C, S725C, and S731C have very low transport activity, implying that this region has a structurally and/or catalytically important role. We measured the residual anion transport activity after mutant treatment with the membrane-impermeant, cysteine-directed compound, sodium (2-sulfonatoethyl)methanethiosulfonate) (MTSES). Only two mutants, S852C and A858C, were inhibited by MTSES, indicating that these residues may be located in a pore-lining region.