Pituitary tumor-transforming gene 1 enhances metastases of cervical cancer cells through miR-3666-regulated ZEB1.

Early cancer metastases often occur in cervical cancer (CC) patients, resulting in poor prognosis and poor therapeutic outcome after resection of primary cancer. Hence, there is a compelling requirement for elucidating the molecular mechanisms underlying the CC cell invasiveness. Recently, the role of microRNAs (miRNAs) and pituitary tumor-transforming gene 1 (Pttg1) in the carcinogenesis of CC has been reported. Nevertheless, the relationship between miRNAs and Pttg1 remains ill-defined. Here, we showed that the levels of miR-3666 were significantly decreased and the levels of zinc finger E-box binding homeobox 1 (ZEB1) and Pttg1 were significantly increased in the CC specimens from patients, compared to the paired non-tumor tissue. Moreover, the levels of miR-3666 and ZEB1 inversely correlated. Bioinformatics analyses showed that miR-3666 targeted the 3'-untranslated region (3'-UTR) of ZEB1 messenger RNA (mRNA) to inhibit its translation, which was confirmed by luciferase reporter assay. Moreover, Pttg1 overexpression inhibited miR-3666 and subsequently increased ZEB1 and cell invasion, while Pttg1 depletion increased miR-3666 and subsequently decreased ZEB1 and cell invasion. Together, our data suggest that Pttg1 may increase CC cell metastasis, possibly through miR-3666-regulated ZEB1 levels.

Design of Machine Learning Algorithms and Internal Validation of a Kidney Risk Prediction Model for Type 2 Diabetes Mellitus.

Objective: This study aimed to explore specific biochemical indicators and construct a risk prediction model for diabetic kidney disease (DKD) in patients with type 2 diabetes (T2D). Methods: This study included 234 T2D patients, of whom 166 had DKD, at the First Hospital of Jilin University from January 2021 to July 2022. Clinical characteristics, such as age, gender, and typical hematological parameters, were collected and used for modeling. Five machine learning algorithms [Extreme Gradient Boosting (XGBoost), Gradient Boosting Machine (GBM), Support Vector Machine (SVM), Logistic Regression (LR), and Random Forest (RF)] were used to identify critical clinical and pathological features and to build a risk prediction model for DKD. Additionally, clinical data from 70 patients (nT2D = 20, nDKD = 50) were collected for external validation from the Third Hospital of Jilin University. Results: The RF algorithm demonstrated the best performance in predicting progression to DKD, identifying five major indicators: estimated glomerular filtration rate (eGFR), glycated albumin (GA), Uric acid, HbA1c, and Zinc (Zn). The prediction model showed sufficient predictive accuracy with area under the curve (AUC) values of 0.960 (95% CI: 0.936-0.984) and 0.9326 (95% CI: 0.8747-0.9885) in the internal validation set and external validation set, respectively. The diagnostic efficacy of the RF model (AUC = 0.960) was significantly higher than each of the five features screened with the highest feature importance in the RF model. Conclusion: The online DKD risk prediction model constructed using the RF algorithm was selected based on its strong performance in the internal validation.

Pretreatment with anti-flagellin serum delays acute lung injury in rats with sepsis.

OBJECTIVE: This study examined the reduction of sepsis-induced ALI by inhibition of flagellin-stimulated TLR5 signaling. METHODS: Rats were randomly divided into three groups: one group served as the sham-operated group (control group), and the other two groups received the induction of sepsis (sepsis and treatment groups). The treatment group was injected with anti-flagellin serum before induction of sepsis. At 2, 4, 6, 12, 24, and 48 h following induction of sepsis (six time-point subgroups, n = 10 per subgroup), arterial PaO(2), wet/dry (W/D) lung weight ratios, levels of serum and BALF flagellin and TNF-α, pulmonary pathological alterations, and TLR5 mRNA expression in the lungs were examined. RESULTS: Compared to sham-operated rats, septic rats had: increased levels of serum and BALF flagellin at 6, 12, 24, and 48 h; reduced arterial PaO(2); elevated W/D lung weight ratio; increased serum and BALF TNF-α levels; and up-regulated TLR5 mRNA expression at 12, 24, and 48 h (P < 0.01). Pretreatment with anti-flagellin serum, however, significantly inhibited sepsis-associated declines in arterial PaO(2), increased W/D lung weight ratios, elevated serum and BALF TNF-α levels, and up-regulated TLR5 mRNA expression at 24 and 48 h (P < 0.01). CONCLUSION: Neutralizing the actions of circulating flagellin with anti-flagellin serum delayed the development of ALI in rats with sepsis.

Multiple skin abscesses associated with bacteremia caused by Burkholderia gladioli: A case report.

BACKGROUND: Burkholderia gladioli (B. gladioli) is regarded as a rare opportunistic pathogen. Only a few patients with abscesses caused by B. gladioli infections have been reported, and these are usually abscesses at the incision caused by traumatic surgery. CASE SUMMARY: A 74-year-old male patient with abscesses and pain throughout his body for 1 mo was admitted to our hospital. Some of the abscesses had ruptured with purulent secretions on admission. Color Doppler ultrasound examination of the body surface masses showed mixed masses 75 mm × 19 mm, 58 mm × 17 mm, 17 mm × 7 mm, and 33 mm × 17 mm in size in the muscle tissues of both the right and left forearms, the posterior area of the right knee and the left leg, respectively. Abscess secretions and blood cultures grew B. gladioli. The following 3 methods were used to jointly identify the bacterium: an automatic microbial identification system, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and full-length 16S rDNA sequencing. After 27 d of treatment with meropenem, etimicin, trimethoprim-sulfamethoxazole and other antibiotics, most of his skin abscesses were flat and he was discharged without any symptoms. CONCLUSION: This is the first reported case of multiple skin abscesses associated with bacteremia caused by B. gladioli. Our study provides important reference values for the clinical diagnosis and treatment of B. gladioli infections.

Cell-surface nucleolin is involved in lipopolysaccharide internalization and signalling in alveolar macrophages.

C23 (nucleolin) shuttling between the nucleus, cytoplasm and cell surface has been implicated in controlling regulatory processes and may play a role in pathogen infection and autoimmune diseases. It has been reported that cell surface-expressed C23 on THP-1 monocytes is involved in the inflammatory response induced by LPS (lipopolysaccharide). This study investigates whether C23 is a membrane receptor for LPS during LPS-induced AMs (alveolar macrophages) activation. First, using immunofluorescence and microscopy, we detected the expression of C23 on the surface of AMs. Second, using LPS affinity columns, we demonstrated that C23 directly binds to LPS. Third, we found that LPS colocalized with C23 on both the cell surface and in the cytoplasm. Finally, knockdown of C23 expression on the cell surface using siRNA (small interfering RNA) led to significant reductions in the internalization of LPS, in LPS-induced NF-κB (nuclear factor κB)-DNA binding and in the protein expression of TNF (tumour necrosis factor)-α and IL-6 (interleukin-6). These findings provide evidence that cell-surface C23 on AMs may serve as a receptor for LPS and are essential for internalization and transport of LPS. Furthermore, C23 participates in the regulation of LPS-induced inflammation of AMs, which indicates that cell-surface C23 is a new and promising therapeutic target for the treatment of bacterial infections.

A meta-analysis of randomized controlled trials in targeted treatments of chronic thromboembolic pulmonary hypertension.

BACKGROUND: Chronic thromboembolic pulmonary hypertension (CTEPH) results in a progressively worsening course associated with substantial morbidity and mortality. The purpose of this comprehensive study was to determine the clinical efficacy of targeted therapeutic interventions for this disease. METHODS: We searched Medline, Embase, Cochrane databases and Pubmed for relevant clinical studies. Randomized controlled trials comparing the effects of targeted treatments to control in CTEPH population were included. Pooled estimates were calculated using a random effect model. Heterogeneity was determined using the I2 statistic. RESULTS: This analysis included 6 studies with a total of 565 patients. We found that targeted treatments approved for pulmonary arterial hypertension (PAH) were associated with a larger improvement in exercise capacity, haemodynamic parameters, functional status and clinical symptom. There were no statistically significant differences associated with targeted treatments compared with control in all-cause mortality and safety outcomes. CONCLUSIONS: This is the first systematic review and meta-analysis of randomized controlled trials revealing a positive role of PAH-targeted therapies in CTEPH. Future larger randomized trials with a focus on long-term clinical outcomes are urgently needed.

[The expression of scavenger receptor class A in the lung in mice with acute lung injury induced by lipopolysaccharide].

OBJECTIVE: To investigate the expression of scavenger receptor class A (SR-A) in lung tissue and alveolar macrophage (AM) in mice with acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: The model of ALI was reproduced by intraperitoneal (ip) injection of LPS (5 mg/kg), and the expression of SR-A was observed in lung tissue and AM, and the expression and distribution of J774 A.1 were also studied by using mouse macrophage line. RESULTS: Partial pressure of oxygen in artery (PaO(2)) of lung tissue in LPS groups was significantly lower while wet/dry weight (W/D) ratio was higher than those in normal saline control group (both P<0.01). SR-A was widely expressed in murine lung, including AM, pulmonary vascular endothelial cells, vascular smooth muscle cells, lung epithelial cells and polymorphonuclear neutrophils. Immunohistochemical staining showed that, during the course of development of ALI in mice, the expression of SR-A was gradually down-regulated with the elapse of time after ip injection of LPS, The same result was seen in the isolated AM, and especially marked in group 4 hours and group 8 hours. In vitro, the expression of SR-A on J774 A.1 cells decreased was also down-regulated when treated with LPS. CONCLUSION: SR-A was widely expressed in murine lung. During the course of ALI, the expression of SR-A in the murine lung and AM is down-regulated, which may be induced by LPS.

Establishment of age- and gender-specific pediatric reference intervals for liver function tests in healthy Han children.

BACKGROUND: The development and growth of children influence values of liver function tests. This study aims to establish age- and gender-specific pediatric reference intervals of liver function among Han children in Changchun, China. METHODS: A total of 1394 healthy Han children, aged 2-14 years, were recruited from communities and schools with informed parental consent in Changchun. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), alkaline phosphatase (ALP), total protein (TP), albumin (ALB), total bilirubin (TBIL) and direct bilirubin (DBIL) were measured on Hitachi 7600-210 automatic biochemical analyzer. The age- and gender-specific reference intervals were partitioned using Harris and Boyd's test and calculated using nonparametric rank method. The pediatric reference intervals were validated in five representative hospitals located in different areas in Changchun. RESULTS: All the analytes required some levels of age partitioning. Proteins (TP, ALB) and bilirubins (TBIL, DBIL) required no gender partitioning. In contrast, considerable gender partitioning was required for serum ALT, AST, GGT, and ALP. TP, TBIL, and DBIL showed steady increases, and AST showed apparent decreases over time, whereas ALT, GGT, ALP, and ALB demonstrated complex trends of change. ALT and GGT increased sharply in males from 11 to 14 years old. However, ALP declined in females from 13 to 14 years. All five laboratories passed the validation of reference intervals. CONCLUSIONS: There were apparent age or gender variations of the reference intervals for liver function. When establishing pediatric reference intervals, partitioning according to age and gender is necessary.

Statins may be beneficial for patients with pulmonary hypertension secondary to lung diseases.

BACKGROUND: Previous animal studies and clinical trials report inconsistent findings regarding the role of statins in pulmonary hypertension (PH). Systematic reviews have shown no use of statins on pulmonary arterial hypertension (PAH). This is the first meta-analysis of randomized controlled trials (RCTs) determining the clinical impacts of statin therapy on patients with PH secondary to lung diseases. METHODS: Electronic databases and manual bibliographical searches were conducted. Eligible studies included RCTs of at least 3 months that evaluated statin therapy as compared with control in adult patients with PH due to pulmonary diseases. Statistical analyses were performed to calculate mean difference, relative risks (RRs), and 95% confidence intervals (CIs) using random-effect model. RESULTS: A total of 6 RCTs were identified and included in this study. Five trials reported the effects of statins in patients with both chronic obstructive pulmonary disease (COPD) and PH, and the remaining 1 was based on PH due to pneumoconiosis. We found that statin therapy was associated with increased 6-minute walk distance and reduced pulmonary artery systolic pressure. There was no observed difference in the incidence of death, drug withdrawal, and adverse event between statin and control group. CONCLUSIONS: Our findings suggest that statins might be safe and beneficial for patients with PH due to chronic lung diseases. However, larger RCTs with more patients and longer observational duration are needed.

Macrophage migration inhibitory factor triggers vascular smooth muscle cell dedifferentiation by a p68-serum response factor axis.

AIMS: Macrophage migration inhibitory factor (MIF) is an important proinflammatory mediator linked to arterial diseases. Although its inflammatory property such as macrophage recruitment is known for contributing to vascular pathogenesis, the direct effects of MIF on homeostasis and biological function of vascular smooth muscle cell (VSMC) that are crucial for development of arterial abnormalities, are poorly understood. METHODS AND RESULTS: We show that MIF is able to directly induce VSMC dedifferentiation, a pathophysiological process fundamental for progression of various arterial diseases. Mechanistically, MIF suppresses p68 protein, a crucial regulator of cell growth and organ differentiation, via activation of JNK and p38 MAPKs. siRNA targeting of p68 facilitated dedifferentiation state in VSMCs, whereas p68 overexpression blocked MIF-elicited transition. In addition, MIF decreased the expression of serum response factor (SRF) that governs VSMC differentiation marker genes transcription, through repression of p68 protein. Furthermore, we showed a previously uncharacterized molecular interaction between p68 and SRF by co-immunoprecipitation assay. p68 attenuated MIF-elicited suppression of SRF recruitment to VSMC-specific promoter. Finally, anti-MIF treatment could reverse VSMC dedifferentiation, preserve vascular function, and inhibit remodelling due to vascular injury. CONCLUSIONS: Our results demonstrate a novel mechanism for the regulation of VSMC differentiation by MIF involving p68 and SRF. Strategy for targeting of MIF could inhibit aberrant transition of VSMC in cardiovascular pathogenesis, and may be of therapeutic benefit in phenotype-related arterial remodelling.

Polymorphism of SARS-CoV genomes.

In this work, severe acute respiratory syndrome associated coronavirus (SARS-CoV) genome BJ202 (AY864806) was completely sequenced. The genome was directly accessed from the stool sample of a patient in Beijing. Comparative genomics methods were used to analyze the sequence variations of 116 SARS-CoV genomes (including BJ202) available in the NCBI GenBank. With the genome sequence of GZ02 as the reference, there were 41 polymorphic sites identified in BJ202 and a total of 278 polymorphic sites present in at least two of the 116 genomes. The distribution of the polymorphic sites was biased over the whole genome. Nearly half of the variations (50.4%, 140/278) clustered in the one third of the whole genome at the 3' end (19.0 kb-29.7 kb). Regions encoding Orf10-11, Orf3/4, E, M and S protein had the highest mutation rates. A total of 15 PCR products (about 6.0 kb of the genome) including 11 fragments containing 12 known polymorphic sites and 4 fragments without identified polymorphic sites were cloned and sequenced. Results showed that 3 unique polymorphic sites of BJ202 (positions 13 804, 15 031 and 20 792) along with 3 other polymorphic sites (26 428, 26 477 and 27 243) all contained 2 kinds of nucleotides. It is interesting to find that position 18379 which has not been identified to be polymorphic in any of the other 115 published SARS-CoV genomes is actually a polymorphic site. The nucleotide composition of this site is A (8) to G (6). Among 116 SARS-CoV genomes, 18 types of deletions and 2 insertions were identified. Most of them were related to a 300 bp region (27,700-28,000) which encodes parts of the putative ORF9 and ORF10-11. A phylogenetic tree illustrating the divergence of whole BJ202 genome from 115 other completely sequenced SARS-CoVs was also constructed. BJ202 was phylogeneticly closer to BJ01 and LLJ-2004.

[Study on pollution for the photoelectronic material InP].

The mass spectrum analysis of crystal face (100) and (111) and the photoluminescence analysis of crystal face (100) in the photoelectronic material InP were given. The Hall coefficient, charge carrier concentration and Hall mobility were determined. Experimental results indicate that the pollution of silicon is predominant.

[Studies on the expression of mRNA of anti- and pro-inflammatory cytokines in acute lung injury induced by lipopolysaccharide in rat].

OBJECTIVE: To study the role of pro-inflammatory cytokines including tumor necrosis factor-alpha(TNF-alpha), interleukin-1beta(IL-1beta), IL-6, and anti-inflammatory cytokines including IL-4, IL-10, and IL-13 in acute lung injury(ALI). METHODS: Reverse transcription-polymerase chain reaction(RT-PCR) were used to assess the expressions of mRNA of TNF-alpha, IL-1beta, IL-6, IL-4, IL-10 and IL-13 in the lung tissue after lipopolysaccharide challenge. RESULTS: The expression of TNF-alpha mRNA was seen to peak at 1 hour and IL-6 mRNA at 4 hours in lung tissue after the challenge. The highest expression of mRNA of other cytokines, such as IL-1beta, IL-4, IL-10 and IL-13 in lung tissue appeared 2 hours after LPS-induced lung injury. CONCLUSION: TNF-alpha is an early presenting cytokine in ALI, and IL-6 may play an important role in delayed development of ALI. The over-expression of anti-inflammatory cytokines IL-4, IL-10 and IL-13 may act as promoters rather than protectors in amplification of inflammatory process.

Hypoxia-inducible factor 1 promoter-induced JAB1 overexpression enhances chemotherapeutic sensitivity of lung cancer cell line A549 in an anoxic environment.

The presence of lung cancer cells in anoxic zones is a key cause od chemotherapeutic resistance. Thus, it is necessary to enhance the sensitivity of such lung cancer cells. However, loss of efficient gene therapeutic targeting and inefficient objective gene expression in the anoxic zone in lung cancer are dilemmas. In the present study, a eukaryotic expression plasmid pUC57-HRE-JAB1 driven by a hypoxia response elements promoter was constructed and introduced into lung cancer cell line A549. The cells were then exposed to a chemotherapeutic drug cis-diamminedichloroplatinum (C-DDP). qRT-PCR and western blotting were used to determine the mRNA and protein level and flow cytometry to examine the cell cycle and apoptosis of A549 transfected pUC57-HRE-JAB1. The results showed that JAB1 gene in the A549 was overexpressed after the transfection, cell proliferation being arrested in G1 phase and the apoptosis ratio significantly increased. Importantly, introduction of pUC57-HRE-JAB1 significantly increased the chemotherapeutic sensitivity of A549 in an anoxic environment. In conclusion, JAB1 overexpression might provide a novel strategy to overcome chemotherapeutic resistance in lung cancer.

Intravenous delivery of bone marrow-derived endothelial progenitor cells improves survival and attenuates lipopolysaccharide-induced lung injury in rats.

Acute lung injury (ALI) is a devastating disease, which is characterized by diffuse endothelium, epithelial damage, and increased pulmonary capillary permeability. Recent data have suggested that the circulating endothelial progenitor cells (EPCs) play an important role in endothelial repair after vascular injury. This study was undertaken to investigate possible endothelial-repairing effects of EPC transplantation after LPS-induced ALI in rats. Using Y-chromosome in situ hybridization and reverse transcription polymerase chain reaction assay, we detected the expression of sex-determining region y in the injured lungs of female model rats, suggesting that allogenic EPCs can migrate to the injured lung tissues. Rats that have received the EPC treatment had a reduced pulmonary edema level, inflammation, hemorrhage, and hyaline membrane formation, as well as an increased survival rate from 44% to 81%. Furthermore, anti-inflammatory cytokine IL-10 levels were dramatically increased in the EPC-treated rats compared with the phosphate buffered saline-treated rats. On the contrary, endothelin-1 and iNOS were downregulated in the EPC-treated group. These findings provide evidence that i.v. EPC treatment results in engraftment of EPCs to the injured lung tissue, which can significantly attenuate lung injury and improve survival in ALI rats. The beneficial effects of EPC engraftment is likely to come from maintaining the integrity of pulmonary alveolar-capillary barrier, reestablishing the endothelial function in vessels and ameliorating the inflammatory state.

The localization of two mimic epitopes of lipopolysaccharide binding protein and the preparation of their tandem multiple antigen peptide.

OBJECTIVE: To screen and identify two mimic epitopes in the inflammatory site of lipopolysaccharide binding protein (LBP), and to synthesize and purify their corresponding mimic epitope four branched peptide (multiple antigen peptide, MAP). METHOD: Using an anti-full length LBP monoclonal antibody as the target molecule, the amino acid sequences of the exogenous peptides were deduced by combining several different techniques including: affinity screening of the phage display peptide library, the lipopolysaccharide (LPS) binding activity assay and competitive inhibition test, cytokine production inhibition, flow cytometry, and DNA sequencing. Basic Local Alignment Search Tool (BLAST) software was used to compare the resulting peptide sequences with the primary structure sequence of the LBP molecule, and thus the amino acid sequences for two mimic inflammatory epitopes for the binding of LBP and LPS were determined. Additionally, the two target sequences were coupled, and the 9-fluorenylmethyloxycarbonyl (FMOC) solid-phase synthesis method was used to synthesize the 24aa peptide. The design program of the multiple antigen peptide (MAP) was used to couple the four tandem peptides with lysine as the core base to produce the branch like structure, and thus, the four branched peptide was synthesized and purified. RESULTS: Fourteen phage clones (C) with competitive LPS binding activity with LBP were successfully obtained. Among these, the amino acid sequences of the peptides in C2, C19, C57, C77, C85 and C91 showed a homology of more than 90% to the primary structure of LBP. However, the amino acid sequences of C29 and C90, WKAQKRFMKKSG and LKTRKLFWKYKD, respectively, did not show homology to the primary structure of LBP, which were determined to be mimic epitopes of the inflammatory sites in LBP. Further synthesis of the 24aa peptide using FMOC solid-phase synthesis and MAP modification were carried out, the four branched peptide was synthesized and purified, and the purity was found to be higher than 95%. The purified peptide was subjected to mass spectrometry analysis and amino acid analysis, and its molecular weight (3102.77 kDa) and amino acid composition were in accordance with theoretical values. CONCLUSION: The amino acid sequence for two mimic epitopes of the inflammatory site of LBP were determined to be WKAQKRFMKKSG and LKTRKLFWKYKD. The MAP was successfully prepared simultaneously and is able to be used as the core antigen protein for the formulation of vaccines. This knowledge will help in future investigations of the functional characteristics of LBP protein, and enhance exploration into new pathways for the prevention and treatment of LPS inflammatory diseases.

[Studies on the changes in the interleukin-13 expression and on activator protein-1 activity in rat pulmonary tissue with acute lung injury induced by endotoxin].

OBJECTIVE: To investigate the changes in plasma level of interleukin-13 (IL-13) and the changes in the pulmonary IL-13 mRNA content and the pulmonary activator protein-1 (AP-1) activity of the rats inflicted with acute lung injury (ALI) induced by lipopolysaccharide (LPS), so as to explore the relationship between IL-13 expression and AP-1 activity. METHODS: One hundred and twenty Wistar rats were employed in the study and were randomly divided into A (2 mg/kg), B (4 mg/kg), C (6 mg/kg) and D (8 mg/kg) groups according to different dosage of LPS administration and a control group (NS group) at each observing time point. The rats were observed at 1, 2, 4 and 6 postburn hours (PBHs) and every 6 rats were deployed in every group and each time points. A model of systemic inflammatory response syndrome-acute lung injury (SIRS-ALI) was replicated in Wistar rats. The plasma content of IL-13 was assayed by enzyme-linked immunosorbent assay (ELISA), and the pulmonary tissue content of IL-13 mRNA and AP-1 activity by reverse transcriptase-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assays (EMSA). RESULTS: The plasma content of IL-13, pulmonary content of IL-13 mRNA and AP-1 activity increased simultaneously after LPS administration. All the above indices were significantly different statistically between the LPS groups and the control group (P < 0.05 - 0.01). The plasma level of IL-13 and pulmonary tissue mRNA content and AP-1 activity in A, B, C and D groups were increased significantly with peak levels at 2 PBHs. CONCLUSION: The pulmonary AP-1 activity increased with the enhanced expression of IL-13, which was related to the development of SIRS-ALI.