BACKGROUND:Endothelial injury is one of the causes of cardiovascular diseases.Human induced pluripotent stem cells are easy to obtain,have strong differentiation ability,and have less exclusiveness,and their endothelial differentiated cells can be used as ideal cells for cardiovascular disease research. OBJECTIVE:To investigate the effect and mechanism of calycosin on endothelial differentiation of human induced pluripotent stem cells and to provide technical support for microvascular regeneration. METHODS:Human induced pluripotent stem cells were divided into control group and calycosin group(1.25,2.5 μg/mL),and growth factors were added to induce single-layer endothelial differentiation.After the induction of differentiation for 8 days,the positive rate of endothelial cell marker CD144 was detected by flow cytometry.Fluorescent expressions of CD144 and CD31 were detected by the immunofluorescence method.Lentivirus RNAi GFP puromycin was used to silence human-induced pluripotent stem cell Piezo1 mRNA followed by endothelial directed differentiation.After 8 days of differentiation,the positive rate of CD144 in differentiated cells was detected by flow cytometry.The mRNA expression levels of CD144,Piezo1 and MEK were detected by qPCR. RESULTS AND CONCLUSION:(1)Compared with the control group,the positive rate of CD144 was significantly increased in the 1.25 and 2.5 μg/mL calycosin groups(P<0.05).The expressions of CD144,Piezo1,and MEK mRNA were increased in the 2.5 μg/mL calycosin group(P<0.05).The fluorescence expressions of CD144(P<0.01)and CD31(P<0.001)were significantly increased in the 2.5 μg/mL calycosin group.(2)Compared with the shNT group,CD144 positive rate and CD144,Piezo1,MEK mRNA expressions were significantly increased in the shNT + calycosin 1.25,2.5 μg/mL groups(P<0.05).Compared with the shPiezo1 group,the positive rate of CD144 and mRNA expressions of CD144,Piezo1 and MEK had no significant changes in the shPiezo1+calycosin 1.25,2.5 μg/mL groups(P>0.05).(3)It is concluded that 2.5 μg/mL calycosin promotes the differentiation of human-induced pluripotent stem cells into endothelial lineages.Calycosin promotes the downstream MEK expression,thereby promoting the endothelial differentiation of human induced pluripotent stem cells by targeting the expression level of Piezo1.

Background: The gestational risk factors predispose to the manifestation of early childhood atopic dermatitis (AD). Objective: We evaluated the association between modifiable and non-modifiable gestational and prenatal risk factors that affect the AD prevalence in children. Methods: We performed the systematic review and meta-analysis of cohort studies (n=27) in PubMed and EMBASE (2000~2021). A meta-analysis was performed using random-effects models to estimate pooled odds ratios (OR) or hazard ratio (HR). We performed a systematic review according to Preferred Reporting Item for Systematic Review and Meta-Analyses (PRISMA) guidelines and summarized cohort studies investigating gestational and prenatal risk factor those predispose to AD in off spring. Leading modifiable and non-modifiable were identified through ORs. Meta-analysis using the random effect model was also conducted to provide an overall estimate for several significant factors. Results: Among the non-modifiable risk factors gestational diabetes (7.2, 95% confidence interval [CI]: 1.4~34.5), maternal history of allergy (2.14, 95% CI: 1.54~2.97) and prenatal history of eczema (2.46, 95% CI: 1.0~5.8) were found as major determining risk factors in early manifestation of AD in children. Further, maternal exposure to industrial products (1.89, 95% CI: 1.10~3.16), exposure to antibiotics during pregnancy (3.59, 95% CI: 1.19~10.85) and passive smoking during pregnancy (2.60, 95% CI: 1.11~6.1) are leading causes of early AD manifestation. Conclusion Conclusively, both genetic and environmental factors play a pivotal role in early manifestation of AD. The better managing the environmental factors during gestational phase to the least can help curtail the prevalence of AD in children.

Objective To investigate the relationship between procalcitonin (PCT),neutrophil to lymphocyte ratio (NLR) and prognosis of acute pancreatitis.Methods Retrospective analysis was made on the levels of PCT and NLR in 102 patients with acute pancreatitis and the same number of healthy controls who sought routine medical care in the hospital,and comparisons were made between the 2 groups.Then,the relationship between PCT/NLR levels and clinical features of the patients were analyzed.Multivariable logistic regression model and receiver operating characteristic (ROC) were used to evaluate the relationship between NLR/PCT and prognosis of acute pancreatitis.Results The levels of NLR and PCT in the patients with acute pancreatitis were significantly higher than those of the healthy individuals,and both NLR and PCT were closely associated with the patient conditions.The area under curve (AUC) for PCT and NLR were respectively 0.81 and 0.84,and the AUC in multivariable combination usage was 0.96.Conclusion Both NLR and PCT were associated with prognosis of acute pancreatitis,and the combined use of NLR and PCT would facilitate to predict the prognosis of acute pancreatitis.

Objective:To study the effect of different dose of persicae semen extract extract(PSE) to barrier function of the intestinal mucous membrane and immunologic function in acute pancreatitis rats.Methods:A total of 48 rats were divided into model control group,low dose,medial dose and high dose PSE groups,and there were 12 rats in each group.Another 12 rats were Sham-operation group.After anesthesia recovery,rats in low dose,medial dose and high dose PSE groups respectively received PSE 0.12 g/kg,0.248 g/kg and 0.36 g/kg,and rats in Sham-operation group and model control group receive isovolumetric distilled water,once per 6 h,4 times in 24 hours.All rats were anesthetized by 10%chloral hydrate after in 24th hour after dosing.Thorax and enterocoelia were opened; 5 ml of blood were respectively drawed to EDTA-anticoagulation tube and un-anticoagulation tube from aorta abdominalis.CD4+, CD8+and Treg cells were determined by direct fluorescent-labelded flow cytometry.IgA, IgG and IgM were determined by immunoturbidimetry.Serum amylase was determined by EPS-G7 substrate,D-lactic acid was determined by enzymology, and serum diamine oxidase was determined by active ration of colorimetry method.Pathological examination of small intestine mucous membrane tissue was taken after HE staining.sIgA in small intestine was determined by radioimmunoassay.mRNA of TLR4 and NF-κBp65 in small intestine tissue was determined by RT-PCR.Results:(1) Serum amylase,D-lactic acid and diamine oxidase in medial dose and high dose PSE groups were significantly decreased ( P<0.01 ) , and sIgA in small intestine was significantly increased ( P<0.01).These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(2) CD4+and CD4+/CD8+in medial dose and high dose PSE groups were significantly increased(P<0.01),and CD8+,Treg cells were significantly decreased(P<0.01) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(3) IgA,IgG and IgM in medial dose and high dose PSE groups were significantly decreased(P<0.01) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups(P<0.01).(4) Small intestine mucous membrane tissue in Sham-operation group was not damaged significantly,but that in model control group was damaged significantly.Small intestine mucous membrane tissue in low dose PSE group was similar to that in model control group,and damage in medial dose and high dose PSE groups was decreased significantly.( 5 ) mRNA of TLR4 and NF-κBp65 in small intestine tissue in medial dose and high dose PSE groups were significantly increased ( P<0.01 ) compared with those in low dose PSE group.These indicators were significantly different in medial dose and high dose PSE groups ( P<0.01 ).Conclusion: PSE has protective effect to barrier function of the intestinal mucous membrane,and significantly improve the immunologic function.

This study was undertaken to observe the biochemical changes in striatum of Parkinson's disease (PD) rat model by modified proton magnetic resonance spectroscopy. 12 SD rats were divided into model (n=7) and control (n=5) groups. At 3 weeks after the injection of 6-hydroxydopamine into right striatum, 1H-MRS on the striatum was taken by modified proton magnetic resonance spectroscopy, and then tyrosine hydroxylase (TH) immunostatining was used to visualize the changes of the neurons in substantia nigra and neurites in striatum. The results showed that TH positive neurons and neurites in the substantia nigra compacts (SNc) and striatum in the normal side of the rat model of PD were decreased (P < 0.05), which proved the successful establishment of PD models. The NAA/Cr ratio of the injected side striatum of model group was lower than that of the normal side (P < 0.05). The ratios of Cho/Cr showed no significant difference between the two sides (P > 0.05). These results indicated that the modified 1.5T 1H-MRS should be a noninvasive technique which could provide useful information about the biochemical metabolites in striatum for the study of PD in rat model.
Animals ; Corpus Striatum ; enzymology ; Female ; Magnetic Resonance Spectroscopy ; methods ; Male ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tyrosine 3-Monooxygenase ; metabolism

Objective To investigate the expression and clinical significance of Survivin in esophageal car-cinoma.Methods Survivin expression was detected by immunohistochemistry in 61 cases of esophageal squamous carcinoma tissues and 26 tumor-adjacent normal tissues.Results The positive Survivin expression rate was 60.7% (37/61)in tumor tissues.The expression of Survivin was significantly correlated with the tumor grading,infiltration, lymph node metastasis and the survival(P<0.01).Conclusion Survivin may be an important marker for tumor di-agnosis and it is of great important to the active treatment of high-risk patients and to increasing of prognosis values.

Genome of Arabidopsis has a kind of genes encoding proteins with ARM repeat domains and some of these proteins are known to play important roles in plant development and responses to hormone. An Arabidopsis mutant lfr with a distinct phenotype was got in leaf and flower development. The gene is predicted to encode a protein with ARM repeat domains. In order to study its function and molecular mechanism, recombination expression plasmid pGEX-2TGST∶LFR was constructed and transformed into the host bacteria strain Rosetta. Then IPTG was used to induce the recombinant protein expression in engineering strain. The expression products were detected by 12% SDS-PAGE. The GST∶LFR fusion protein was existed in soluble form with a relative molecular mass 77 ku, which is fit with the molecular mass supposed from gene coding frame. After purification by GST-tag affinity chromatography and electroelution, the fusion protein was used as antigen to prepare polyclonal antiserum in rabbits. After the fifth injection of antigen, the antiserum was obtained and further purified by decreased nonspecific bacteria and GST-tag antibody with method of immuno-precipitation. Western blot analysis showed that the purified antiserum, raised against the recombination LFR protein in rabbits, could react to the recombinant protein expressed in Rosetta specifically. And then the nuclear proteins of Arabidopsis wild type and mutant were extracted and separated by SDS-PAGE. Western blot assays revealed that there was a protein band, with a relative molecular mass 50 ku, indicating that antiserum could react to the native protein expressed in Arabidopsis specifically.