Objective: To investigate the effects of glycogen synthase kinase-3β (GSK3β)/eukaryotic extension factor kinase 2 (eEF2K) signaling pathway on the process of pulmonary fibrosis through in vivo experiments, and find new ideas for clinical treatment of pulmonary fibrosis. Methods: The pulmonary fibrosis model of C57BL/6 male mice was induced by bleomycin with intratracheal injection at the dose of 2 mg/kg. After 14 days of modeling, animals were divided into model group, negative inhibition group and inhibition group (n=5 for each group), and control group was not processed. The inhibition group was treated with TDZD-8 (4 mg/kg) after modeling, the negative inhibition group was given DMSO solution after modeling, and the samples were collected after 28 days. Hematoxylin-eosin staining method was used to detect lung fibrosis in mice and scored according to Ashcroft scale. Expression levels of GSK3β, p-GSK3β, eEF2K, p-eEF2K (Ser70, Ser392, Ser470), precursor protein of matrix metalloproteinase-2 (pro-MMP-2), matrix metalloproteinase-2 (MMP-2), collagen I (Col I), collagen Ⅲ (Col Ⅲ) and α-smooth muscle actin (α-SMA) were detected by Western blot. Results: Compared with control group, the fibrosis score was up-regulated, the expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were increased, while that of eEF2K was decreased in model group (P＜0.05). Compared with model group, the fibrosis score, expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were decreased, but the expression level of eEF2K was increased in inhibition group (P＜0.05). Conclusion: GSK3β can activate eEF2K by phosphorylation at the sites of Ser70, Ser392 and Ser470, increase the contents of fibrosis indicators, promote the formation of pulmonary fibrosis, and aggravate lung tissue lesions.
Animals ; Collagen ; Collagen Type I ; Elongation Factor 2 Kinase/metabolism* ; Eukaryota/metabolism* ; Fibrosis ; Glycogen Synthase Kinase 3 beta ; Male ; Matrix Metalloproteinase 2/metabolism* ; Mice ; Mice, Inbred C57BL ; Pulmonary Fibrosis/chemically induced* ; Signal Transduction

BACKGROUND: Delivery room resuscitation assists preterm infants, especially extremely preterm infants (EPI) and extremely low birth weight infants (ELBWI), in breathing support, while it potentially exerts a negative impact on the lungs and outcomes of preterm infants. This study aimed to assess delivery room resuscitation and discharge outcomes of EPI and ELBWI in China. METHODS: The clinical data of EPI (gestational age [GA] <28 weeks) and ELBWI (birth weight [BW] <1000 g), admitted within 72 h of birth in 33 neonatal intensive care units from five provinces and cities in North China between 2017 and 2018, were analyzed. The primary outcomes were delivery room resuscitation and risk factors for delivery room intubation (DRI). The secondary outcomes were survival rates, incidence of bronchopulmonary dysplasia (BPD), and risk factors for BPD. RESULTS: A cohort of 952 preterm infants were enrolled. The incidence of DRI, chest compressions, and administration of epinephrine was 55.9% (532/952), 12.5% (119/952), and 7.0% (67/952), respectively. Multivariate analysis revealed that the risk factors for DRI were GA <28 weeks (odds ratio [OR], 3.147; 95% confidence interval [CI], 2.082-4.755), BW <1000 g (OR, 2.240; 95% CI, 1.606-3.125), and antepartum infection (OR, 1.429; 95% CI, 1.044-1.956). The survival rate was 65.9% (627/952) and was dependent on GA. The rate of BPD was 29.3% (181/627). Multivariate analysis showed that the risk factors for BPD were male (OR, 1.603; 95% CI, 1.061-2.424), DRI (OR, 2.094; 95% CI, 1.328-3.303), respiratory distress syndrome exposed to ≥2 doses of pulmonary surfactants (PS; OR, 2.700; 95% CI, 1.679-4.343), and mechanical ventilation ≥7 days (OR, 4.358; 95% CI, 2.777-6.837). However, a larger BW (OR, 0.998; 95% CI, 0.996-0.999), antenatal steroid (OR, 0.577; 95% CI, 0.379-0.880), and PS use in the delivery room (OR, 0.273; 95% CI, 0.160-0.467) were preventive factors for BPD (all P < 0.05). CONCLUSION Improving delivery room resuscitation and management of respiratory complications are imperative during early management of the health of EPI and ELBWI.
Birth Weight ; Bronchopulmonary Dysplasia ; China/epidemiology* ; Delivery Rooms ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Extremely Premature ; Infant, Newborn ; Male ; Pregnancy

Objective:To investigate the association between sleep quality and the risk of acute exacerbation in mild and moderate chronic obstructive pulmonary disease （COPD） patients in Pudong New Area of Shanghai. Methods:This was a prospective study involving eligible mild and moderate COPD patients from 10 communities randomly selected in Pudong New District of Shanghai. A structured questionnaire was used to collect demographic characteristics， clinical information and information on acute exacerbation. Sleep quality was assessed using the Pittsburgh Sleep Quality Index （PSQI） in Chinese. Multiple negative binomial regression was used to estimate the association between sleep quality and risk of exacerbation. Results:Altogether 212 mild/moderate COPD patients participated and completed the entire survey， of whom the majority （95.8%） were mild COPD patients， 110 persons female and over half （54.2%） over 65 years old. 32.9% of the patients had poorer sleep quality at baseline. 18.9% of the patients reported exacerbation over the past year during follow-ups. Multiple negative binomial regression suggested that increased PSQI was related to higher risk of exacerbation （RR_ad=1.12， 95%CI：1.02-1.24）， and patients with poorer sleep efficiency had a higher risk of exacerbation （RR_ad=1.66， 95%CI：1.17-5.43）. Conclusion:Poorer sleep quality is associated with a higher risk of exacerbation in community mild/moderate COPD patients， especially in those with problem of sleep efficiency. More attention to sleep disorders is warranted in community management or self-management of patients with COPD.

Objective　At present, the clinical significance and biological function of Msi1 (Musashi1) in colon cancer are still not very clear. So, a comprehensive understanding of the expression and role of Msi1 in colon cancer has important clinical and theoretical significance. This study is to investigate the clinical significance of Msi1 gene and its biological role in colon cancer by lentiviral vector to interfere with Msi1 gene expression in colon cancer SW480 cells.　Methods　20 colon cancer specimens were collected from the Second Surgery Department of the Fourth Hospital of Hebei Medical University from October 2013 to May 2014. Each specimen was collected from the cancer tissue and the adjacent intestinal wall tissue. Western blot was performed to determine the protein expression of Msi1 in tumor tissues and adjacent normal tissues from colon cancer patients. The relationship between Msi1 protein expression and clinical characteristics was further analyzed. The lentiviral vector was used to construct a stable SW480 cell line with low expression of Msi1. The lentivirus containing two different interference sequences (shmsi1-1 and shmsi1-2) was transfected into the target cells, and the colon cancer cells were divided into control group (without any treatment), shMsi1-1 group (transfected shMsi1-1) and shMsi1-2 group (transfected shMsi1-2). The two lentivirus silencing effects were detected by Western blot. Cell proliferation was detected by CCK-8 assay, Clone formation assay was conducted to detect the colony forming ability, and Flow cytometry analysis was used to examine the apoptosis rate.　Results　The protein expression of Msi1 in colon cancer tissue(0.863±0.208) was significantly higher than that in adjacent normal tissues(0.272±0.078), and the difference was statistically significant(P<0.001). The relative expression of Msi1 protein in shMsi1-1 and shMsi1-2 groups (0.299±0.111 and 0.207±0.087) was significantly lower than that in the control group (1.000±0.149) (P<0.001). The proliferation rate of shMsi1-1 and shMsi1-2 at 48 h and 72 h was significantly lower than that of the control group (P<0.01). Compared with the control group (296.33±64.04), shMsi1-1 group (92.00±43.31) and shMsi1-2 group (78.67±32.87) were significantly decreased (P<0.01). Compared with the control group [(4.01±0.26) %], the apoptosis rate of shMsi1-1 group, shMsi1-2 group [(10.22±1.04) %, (10.87±1.27) %] was significantly increased (P<0.001).　Conclusion　Interference with Msi1 gene expression inhibits proliferation of colon cancer SW480 cells and promotes tumor cell apoptosis. This finding provides a new intervention target for the clinical treatment of colon cancer.

Objective To explore the association between Toll-like receptors(TLR) gene polymorphisms and the primary immune response level to Hepatitis B Vaccine in Han children in Guangxi. Methods A total of 513 Han children aged 8-9 months were collected from the department of pediatrics in the Maternal and Child Hospital of Guangxi Zhuang Autonomous Region and Nanning Maternal and Child Health Hospital from 2014 to 2016. Peripheral venous blood of each study object was collected to detect HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc and HBV DNA. The polymorphisms of 10 sites of TLR gene were detected by SNPscanTM multiple SNP typing techniques. The association between allele, genotype of TLR gene and anti-HBs levels were analyzed by non-conditional logistic regression. Results The genetic polymorphism of TLR3 gene rs13126816 was related to immune response after primary Hepatitis B immunization in Han children in Guangxi (OR=1.79，95% CI: 1.11-2.89, P=0.018). The anti-HBs level of children with A/A genotype［238.04(519.75) mIU/L］and G/A genotype［347.96(619.68) mIU/L］were significantly lower than those with G/G genotype［489.08(854.76) mIU/L］, and the differences were statistically significant (all P<0.05). Anti-HBs level of children carrying allele A ［317.20(608.72) mIU/L］was significantly lower than those carrying allele G［457.01(852.66) mIU/L］, and the difference was statistically significant (Z=-3.055， P<0.05). The rest of the TLR genes were not related to the immune response of Hepatitis B Vaccine (all P>0.05). Conclusions The allele A of TLR3 gene rs13126816 may be the influencing factor for the low response of primary immune response to Hepatitis B Vaccine in Han children.

By preparing 10 batches of the material reference of Linggui Zhugan Decoction,the methodology of the characteristic spectrum of the material reference was created. The creaming rate range,the contents and the transfer rate range of cinnamaldehyde,glycyrrhizin and glycyrrhizic acid,the characteristic peaks and the similarity range of the characteristic spectrum of Linggui Zhugan Decoction were determined to clarify key quality attributes of the material reference of Linggui Zhugan Decoction. In the 10 batches of the material reference of Linggui Zhugan Decoction,the similarity of characteristic spectrum was higher than 0. 9. Furthermore,after summarizing the characteristic peak information,we knew that Fuling had two characteristic peaks,Guizhi had six characteristic peaks,Baizhu had two characteristic peaks and Gancao had 11 characteristic peaks. The average creaming rate of the material reference of the ten batches was( 12. 13 ± 0. 35) %. The average content of cinnamaldehyde was 0. 32%,the average transfer rate was 10. 69%,the content of cinnamaldehyde in the different batches was between 0. 22% and 0. 42%,and the transfer rate was between 7. 48% and13. 90%. The average content of glycyrrhizin was 0. 84%,the average transfer rate was 50. 39%,the content of glycyrrhizin in the different batches was between 0. 42% and 1. 26%,and the transfer rate was between 35. 27% and 65. 51%. The average content of glycyrrhizic was 1. 88%,the average transfer rate was 40. 74%,the content of glycyrrhizic in the different batches was between 0. 94% and2. 82%,and the transfer rate was between 28. 52% and 52. 96%. In this paper,the quality value transmitting of substance benchmarks of Linggui Zhugan Decoction was analyzed by the combination of characteristic spectrum,creaming rate and the content of index component. A scientific and stable method was preliminarily established,which provided scientific basis for the quality control and formulation development of Linggui Zhugan Decoction.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/standards* ; Glycyrrhiza ; Glycyrrhizic Acid/analysis* ; Plant Extracts/standards* ; Quality Control

Objective To investigate the relationship between the expression of linc-VLDLR in extracellular vesicles (EVs) and the development and drug resistance of esophageal carcinoma.Methods Fifty percent of inhibitory concentration (ICs0) ofadriamycin (ADM) for Eca109 cells was detected by MTT assay,after the treatment of esophageal squamous cell carcinoma Eca109 cell line with different concentrations of ADM for 24h.The culture medium was treated with 3 concentrations of ADM based on the IC50 for 24h for extracting EVs in Eca109 cells.linc-VLDLR mRNA expression in EVs was detected by qRT-PCR assay.ICs0 of ADM for Eca109 cells intervened by EVs for 48h was detected by MTT assay.Cell cycle was detected by FCM and linc-VLDLR and ABCG2mRNA expressions in Eca109 cells were detected by qRT-PCR after the treatment of the EVs for 48h.Results ICs0 of ADM acting on Eca109 cells for 24h was 0.44 ± 0.02μg/ml,so ADM concentrations of 0.2,0.4,0.8μg/ml were choosed in the following studies.EVs were extracted from the supernatant after the treatment of 0,0.2,0.4,0.8μg/ml ADM for 24h and were labeled as EVs1,2,3 and 4 respectively.LincVLDLR mRNA expression in EVs4 was significantly higher than that in EVs1-3 (P＜0.01).ADM ICs0 for Eca109 cells in EVs4 group was significantly higher than that in other groups after the treatment of EVs1-4 on Eca109 cells for 48h (P＜0.05).Flow cytometry results showed that the proliferation index of Eca109 cells in EVs4 group was significantly higher than that in EVs 1-3 and control groups (P＜0.01).Linc-VLDLR and ABCG2 mRNA expression levels in Eca109 cells of EVs4 group were significantly higher than these of EVs1-3 and control groups (P＜0.05).Conclusions High expression of linc-VLDLR and ABCG2 gene in esophageal cancer cells is involved in the formation of esophageal cancer resistance.EVs released by drug-resistant cells can upregulate the expression of ABCG2 in esophageal cancer cells and regulate the drug resistance of esophageal cancer cells,which is related to the linc-VLDLR gene carried by EVs.

OBJECTIVETo investigate the clinical features and prognosis of malignancy-associated hemophagocytic lymphohistiocytosis (MAHS) in children.

METHODSA retrospective analysis was performed for the primary diseases, clinical features, and prognosis of 24 children with MAHS.

RESULTSAmong the 24 children, 11 (46%) had MAHS induced by tumor and 13 (54%) had chemotherapy-associated MAHS. As for primary diseases, 17 children had acute leukemia, 6 had lymphoma, and 1 had neuroblastoma. The most common clinical manifestations were pyrexia, respiratory symptoms, and hepatosplenomegaly. The most common laboratory abnormalities were hemocytopenia, elevated serum ferritin, and elevated lactate dehydrogenase. Of the 24 children, 22 were treated according to the HLH-2004 protocol and 2 gave up treatment; 18 children died, 1 was lost to follow-up, and 5 survived. The survival time ranged from 3 days to 2 years and 4 months (median 28 days).

CONCLUSIONSChildren with MAHS have various clinical features and extremely poor treatment outcomes.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; mortality ; therapy ; Male ; Neoplasms ; complications ; Prognosis ; Retrospective Studies ; Treatment Outcome

Inflammation is one of the important risk factors of rheumatic diseases. Aconiti Radix is widely used for the treatment of rheumatism, which has significant anti-inflammatory effects. However, its anti-inflammatory mechanism on molecular level is still not clear. The purpose of this study is to illuminate the anti-inflammatory mechanism of Aconiti Radix based on the protein interaction network (PIN) analysis on molecular network level. The main anti-inflammatory components (aconitine, hypaconitine and mesaconitine) were chosen in this study to obtain the targets of the components and protein-protein information though databases retrieval and construct the PIN of Aconiti Radix. By a graph theoretic clustering algorithm molecular complex detection(MCODE), 13 modules were identified and analyzed by gene ontology(GO) enrichment. The results showed that the anti-inflammatory mechanism of Aconiti Radix was mainly associated with prostanoid metabolic process and leukocyte chemotaxis mediated by chemokines. In this study, the anti-inflammatory mechanism of Aconiti Radix was elucidated systematically from molecular network level, which provided the scientific basis for the treatment of rheumatic diseases.

OBJECTIVETo analyze the phenotype-genotype correlation of MYH7-V878A mutation.

METHODSExonic amplification and high-throughput sequencing of 96-cardiovascular disease-related genes were carried out on probands from 210 pedigrees affected with hypertrophic cardiomyopathy (HCM). For the probands, their family members, and 300 healthy volunteers, the identified MYH7-V878A mutation was verified by Sanger sequencing. Information of the HCM patients and their family members, including clinical data, physical examination, echocardiography (UCG), electrocardiography (ECG), and conserved sequence of the mutation among various species were analyzed.

RESULTSA MYH7-V878A mutation was detected in five HCM pedigrees containing 31 family members. Fourteen members have carried the mutation, among whom 11 were diagnosed with HCM, while 3 did not meet the diagnostic criteria. Some of the fourteen members also carried other mutations. Family members not carrying the mutation had normal UCG and ECG. No MYH7-V878A mutation was found among the 300 healthy volunteers. Analysis of sequence conservation showed that the amino acid is located in highly conserved regions among various species.

CONCLUSIONMYH7-V878A is a hot spot among ethnic Han Chinese with a high penetrance. Functional analysis of the conserved sequences suggested that the mutation may cause significant alteration of the function. MYH7-V878A has a significant value for the early diagnosis of HCM.

Adult ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Cardiac Myosins ; genetics ; Cardiomyopathy, Hypertrophic ; genetics ; Female ; Genetic Association Studies ; methods ; Genotype ; Humans ; Male ; Middle Aged ; Mutation ; genetics ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Young Adult