Inhibition of abnormal GRK2 transmembrane transfer to reduce maturation of dendritic cells in CIA mice by regulating JAK1-STAT1 signaling pathway / 中国药理学通报

Institute of Clinical Pharmacology of Anhui Medical University, Key Laboratory of And-inflammatory and Immune Medicine, Ministry of Education .Anhui Collaborative Innovation Center of And-Inflammatory and Immune Medicine, Rheumatoid Arthritis Research Center of Anhui Medical University, Jlefei ,230032, China,Aim To reveal the role of the abnormal activation of G protein-coupled receptor kinase 2(GRK2)and abnormal signal transduction of JAK1-STAT1 in the abnormal immune response of rheumatoid arthritis(RA)by exploring the effects of GRK2 on the JAK1-STAT1 signaling pathway in dendritic cells(DCs)of collagen-induced arthritis(CIA)mice and the undelying mechanisms,so as to provide a basis for revealing the new mechanism of RA.Methods The CIA model was established,and the co-stimulatory molecular level of DCs was detected by flow cytometry,the cytokine levels of plasma in mice were detected by ELISA,and the expression of p-JAK1,p-STAT1 and GRK2 in spleen tissues was detected by immunohistochemistry.Bone marrow cells were induced into DCs in vitro and stimulated with IFN-α and PGE2 for 48 h.Flow cytometry was used to detect the level of co-stimulatory molecules and phagocytosis of DCs,and ELISA to detect the level of cytokines in cell supernatant.CO-IP was employed to detect the co-localization of GRK2 and JAK1 in DCs.Western blotting was used to detect the expression of JAK1-STAT1 and the cell membrane expression of GRK2.Imaging flow cytometry was applied to detect the nucleation rate of p-STAT1.Results In vivo the level of co-stimulatory molecules of dendritic cells of CIA mouse increased,and the expression of GRK2 and p-JAK1,p-STAT1 in spleen was positively correlated.The co-localization of GRK2 and JAK1 in spleen of the CIA group decreased significantly.In vitro GRK2 inhibitors reduced the level of costimulatory molecules,cytokines IL-6 and TNF-α,the expression of JAK1 and STAT1,the expression of GRK2 in the cell membrane,and the rate of p-STAT1 nuclear translocation,and increased the Ag uptake capacity of DCs and the co-localization rate of GRK2 and JAK1.Conclusions The abnormal GRK2 transfer to the cell membrane in DCs mediates the maturation of DCs and the activation of the JAK1-STAT1 signaling pathway.Inhibition of GRK2 transfer membrane can restore its control of the JAK1-STAT1 signal transduction of DCs,reduce the maturation of DCs,and play an important role in improving mouse CIA.

Risk factors for the first ventilator weaning failure in preterm infants receiving invasive mechanical ventilation / 中国当代儿科杂志

OBJECTIVE@#To study the risk factors for the first ventilator weaning failure and the relationship between the weaning failure and prognosis in preterm infants receiving invasive mechanical ventilation.@*METHODS@#A retrospective analysis was performed for the preterm infants who were admitted to the Neonatal Intensive Care Unit of Peking University Third Hospital and received mechanical ventilation within 72 hours after birth. According to whether reintubation was required within 72 hours after the first weaning, the infants were divided into a successful weaning group and a failed weaning group.@*RESULTS@#A total of 282 preterm infants were enrolled, and there were 43 infants (15.2%) in the failed weaning group. Compared with the successful weaning group, the failed weaning group had significantly lower gestational age and birth weight (@*CONCLUSIONS@#Use of ≥ 2 vasoactive agents before ventilator weaning and PDA (≥ 2.5 mm) are risk factors for ventilator weaning failure, and ventilator weaning failure may be associated with adverse outcomes in hospitalized preterm infants.

Metabolomic screening for diagnostic biomarkers of drug-induced chronic liver injury related cirrhosis / 药学学报

About 15%-20% of drug-induced liver injury (DILI) will progress to chronic manifestation (CH-DILI), which sometimes advances rapidly to liver cirrhosis (LC-DILI) within 0.5-1 year with deteriorative clinical prognosis. Therefore, it is important to find a non-invasive diagnosis for early detection of liver cirrhosis. In this study, the metabolomic profiles revealed significant differences in the metabolites from the plasma of LC-DILI versus CH-DILI. We found 35 differential metabolites through principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Through pathway enrichment analysis, some up-regulated metabolic pathways reflected impaired liver functions such as bile acid, lipid synthesis and decomposition during cirrhosis. Five biomarkers were found to exhibit effective diagnosis value (AUC > 0.6), including phosphatidylcholine, lysoPC (18:1 (9Z)), creatine, taurochenodeoxycholic acid and taurocholic acid. Furthermore, we found that the relative content ratio between phosphatidylcholine and lysoPC (18:1 (9Z)) had a better distinguishing ability (AUC = 0.867). The relative content ratio also had the feature to reduce systematic errors of sample processing and instrument detection, therefore having a greater value for clinical application.

Effects of extracellular matrix stiffness and thickness on cell growth / 中国组织工程研究

BACKGROUND: The mechanical properties of extracellular matrix can affect cell spreading, proliferation and differentiation. It is of extremely vital significance for tissue engineering and regenerative medicine. OBJECTIVE: To summarize the progress of matrix mechanical properties influences on cell growth and to analyze if there is a relationship between matrix thickness and stiffness, in order to provide experimental methods for functional tissue construction in vitro. METHODS: The author performed a data retrieval of PubMed and Bailianyun databases from 1988 to 2016 to search the articles addressing the influence of extracellular matrix mechanical properties on cell growth, and systematically reviewed the literatures. RESULTS AND CONCLUSION: A total of 254 references were retrieved, and 43 articles were finally involved in the result analysis according to the inclusion and exclusion criteria. After summarizing and analyzing, it is found that the structure and function of cells adhering to the matrix are related to the extracellular environment.In vitro simulation experiments have shown that the increasing of matrix stiffness can change the cell morphology, increase the spreading area and proliferation rate, and can also influence the cell differentiation. When the matrix thickness is in a certain range, cells can sense the change of matrix thickness. When the matrix thickness is out of range, cells cannot perceive the matrix thickness. Matrix stiffness and cross-linking degree have certain effects on the cells, and the matrix thickness ranges from several microns to 60 microns, which is associated with the horizontal size of seeded cells.

Quercetin inhibits proliferation of human cervical carcinoma HeLa cells through affecting tryptophan metabolism / 中国药理学通报

Aim To investigate the mechanism of inhibition on the proliferation of human cervical carcinoma HeLa cells by quercetin.Methods Cell proliferation was detected by CCK-8 method.Flow cytometry was used to detect the cell cycle changes.OD480 values which reflected the metabolism of tryptophan and the production of kynurenine were measured by UV-Vis spectrophotometer.The mRNA levels of IDO1 were detected by qPCR.The expression and purification of IDO1 protein were detected by Western blot.Enzyme activity reaction was performed in vitro,and the content changes of tryptophan and kynurenine were detected by HPLC.Results Quercetin inhibited the proliferation of HeLa cells and led to cell cycle arrest.Quercetin inhibited the metabolism of tryptophan.Quercetin significantly inhibited the enzymatic activity of IDO1 in vitro,but did not affect the expression of IDO1.The addition of kynurenine could reverse the inhibition of cell proliferation induced by quercetin.Conclusion Quercetin affects tryptophan metabolism through inhibiting the enzymatic activity of IDO1.This may be one of the mechanisms by which quercetin exerts its effect on the proliferation of HeLa cells.

Evaluation of quality of life of children with atopic dermatitis before and after treatment / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the quality of life of children with atopic dermatitis (AD) and their families, and to assess the changes in quality of life after treatment.</p><p><b>METHODS</b>The Infants' Dermatitis Quality of Life Index (IDQOL), Children's Dermatology Life Quality Index (CDLQI), and Dermatitis Family Impact (DFI) questionnaires were used to evaluate quality of life in 109 children with AD and 55 normal children. The Severity Scoring of Atopic Dermatitis (SCORAD) was used to evaluate disease severity. The children were given external application of glucocorticoids according to the SCORAD index, and the clinical outcome and changes in quality of life were observed after 3 months of treatment.</p><p><b>RESULTS</b>The three items in both IDQOL and CDLQI questionnaires with higher scores were itching/scratching, mood problems, and sleeping disturbance in the AD patients. Sleeping disturbance, fatigue and mood problems were the three items in the DFI questionnaire with higher scores. There was a positive correlation between IDQOL/CDLQI score and SCORAD index (r=0.358, 0.386 respectively; P<0.05). In the younger group (1-4 years), there was a positive correlation between DFI score and SCORAD index (r=0.297; P<0.05). After treatment the severity of AD and quality of life in the children and their families (P<0.05) were significantly improved.</p><p><b>CONCLUSIONS</b>AD has an adverse effect on quality of life in children with AD and their families. Topical glucocorticoids may control the symptoms of AD and improve the quality of life in children and their families.</p>

5589del8: the recurrent mutation of BRCA1 gene in Chinese breast cancer patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the "hot spot" of BRCA1/2 gene mutations in Chinese mainland breast cancer population.</p><p><b>METHODS</b>The known BRCA1/2 gene mutations in author's previous studies were reanalyzed by denaturing high performance liquid chromatography and DNA sequencing method in 177 patients with early onset breast cancer or affected relatives and 426 sporadic breast cancer patients from four breast cancer centers in China.</p><p><b>RESULTS</b>Three cases were found with BRCA1 5589del8 mutation out of 247 hereditary-predisposing breast cancer patients (70 patients in previous study and 177 patients in current study) and 2 cases with BRCA1 5589del8 mutation out of 426 sporadic breast cancer patients. They had similar even same haplotype.</p><p><b>CONCLUSION</b>BRCA1 5589del8 mutation is likely to be the "founder mutation" in Chinese population, but it should be confirmed by further studies.</p>

BRCA1 germ line mutations in Chinese early-onset breast cancer patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the role of disease associated germ line mutations in BRCA1 gene among Chinese early-onset breast cancer patients.</p><p><b>METHODS</b>A total of 188 early-onset breast cancer patients, who were diagnosed with breast cancer before 41-year-old, were enrolled from four breast cancer clinical centers in China. Thirty-nine of them (20.7%) also had family history of breast/ovarian cancer. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR) for the entire exons and the splicing sites of BRCA1. Twenty-two of the patients were screened by single strand conformation polymorphism (SSCP), and the other 166 of them were screened by denaturing high performance liquid chromatography (DHPLC). The abnormal fragments recognized were ascertained by DNA direct sequencing. For those samples with the same recurrent mutation, five BRCA1-linked markers (D17S855, D17S1322, D17S1323, D17S1326 and D17S1327) were used for the allelotype analysis.</p><p><b>RESULTS</b>Twelve disease-associated mutations were identified in 15 (8.0%) patients, among which BRCA1 1100delAT and 5589del8 were identified in 3 and 2 patients respectively. Nine (23.1%) of them were identified in those with breast/ovarian cancer family history. The difference of BRCA1 mutation frequency between the patients with and without family history was statistically significant (P=0.001). Allelotype analysis showed the two BRCA1 5589del8 mutation carriers shared the same allelotype in all the 5 STR sites, and two of the three 1100delAT mutation carriers, who came from the northern China, also shared the same allelotype in all the 5 STR sites, which were different from those of the 5589del8 mutation carriers'.</p><p><b>CONCLUSION</b>This is a relatively very large scale multi-hospital-based study of BRCA1 mutations in Chinese early-onset breast cancer patients up to now. It seems reasonable to give genetic consultations and genetic test of BRCA1 gene to early-onset breast cancer patients in China, especially for those with breast/ovarian cancer family history. The two recurrent mutations might be founder mutations of Chinese population. It might be cost-effective to analyze these two mutations before whole gene analysis.</p>

A genetic study on microdeletion of azoospermia factor region on Y chromosome of azoospermia and oligozoospermia patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the genetic causes of azoospermia and severe oligozoospermia.</p><p><b>METHODS</b>Cytogenetic analysis and multiplex polymerase chain reaction(PCR) analysis were done on the 148 patients with azoospermia and serious oligozoospermia.</p><p><b>RESULTS</b>Eleven of the 148(7.4%) cases showed microdeletion of at least one STS. In fifteen STS of AZFa, AZFb,AZFd, AZFc, thirteen STS, eleven STS,two STS and one STS microdeletion were found in each case respectively, including two with 12 STS, five with 5 STS microdeletion.Seven cases had chromosomal morphologic changes(4.7%),four had deletion and one had deletion with translocation of long arm on Y chromosome. One had enlarged region one band two(q12) on long arm of Y chromosome and one had reciprocal translocation of autosomes.</p><p><b>CONCLUSION</b>The findings indicated that AZF microdeletion and chromosomal abnormality should be important causes of male infertility.</p>

Using multiplex PCR to analyze the breakpoint of a severe Y-chromosome deletion / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To elucidate the relationship between azoospermia factor(AZF) microdeletion of Y chromosome and azoospermia, the exact breakpoint of a severe Y-chromosome deletion was determined according to the physical map of AZF.</p><p><b>METHODS</b>Multiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY82, sY84, sY86 in AZFa, sY124, sY127, sY128, sY133, sY134, sY143 in AZFb, sY239, sY242 sY254, sY255 in AZFc, and sY145, sY152 in AZFd; sex-determining region Y(SRY) was taken as an internal control. And then sY82,sY86,sY85,sY84 were further analyzed using the sample of the patient who had Y-chromosome deletion by G band analysis to map the breakpoint at molecular level.</p><p><b>RESULTS</b>All 15 STS and sY85 were amplified in positive control while only sY82, sY86 were amplified in the clinical sample, thus the breakpoint was found to be between sY86 and sY85.</p><p><b>CONCLUSION</b>This study on the patient provided the direct biomolecular evidence of the exact breakpoint of the severe Y-chromosome deletion and established the deletion map of acrocentric chromosome. It also proved that the patient's azoospermia was due to the deletion of AZF.</p>

A cytogenetic and molecular genetic study on microdeletion of AZF region on Y chromosome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the morphology of Y chromosome and microdeletion of the correlated specific azoospermia factor(AZF) region on Y chromosome in cases of azoospermia and to identify the genetic diagnosis made for male infertility patients.</p><p><b>METHODS</b>Peripheral blood samples were taken from two patients with azoospermia, and then were examined by use of G banding, C banding cytogenetic analysis and multiplex polymerase chain reaction (PCR) microdeletion analysis.</p><p><b>RESULTS</b>The karyotypes of the two cases were 45, X, -Y, -22, +der(Y)t(Y;22)(q11.2;q11.2) and 46, XY, del(Y)(q11.2) respectively. In 12 sequence-tagged sites(STS) of AZFa, AZFb, AZFd, AZFc, only one was detected in the first case and two were detected in the other case.</p><p><b>CONCLUSION</b>The cytogenetic analysis and the detection of AZF microdeletion on Y chromosome are essential to the final genetic diagnosis to be made for male infertility patients.</p>