[Assessment of risk factors for bronchopulmonary dysplasia with pulmonary hypertension and construction of a prediction nomogram model].

Objective: To explore the risk factors of pulmonary hypertension (PH) in premature infants with bronchopulmonary dysplasia (BPD), and to establish a prediction model for early PH. Methods: This was a retrospective cohort study. Data of 777 BPD preterm infants with the gestational age of <32 weeks were collected from 7 collaborative units of the Su Xinyun Neonatal Perinatal Collaboration Network platform in Jiangsu Province from January 2019 to December 2022. The subjects were randomly divided into a training cohort and a validation cohort at a ratio of 8∶2 by computer, and non-parametric test or χ2 test was used to examine the differences between the two retrospective cohorts. Univariate Logistic regression and multivariate logistic regression analyses were used in the training cohort to screen the risk factors affecting the PH associated with BPD. A nomogram model was constructed based on the severity of BPD and its risk factors,which was internally validated by the Bootstrap method. Finally, the differential, calibration and clinical applicability of the prediction model were evaluated using the training and verification queues. Results: A total of 130 among the 777 preterm infants with BPD had PH, with an incidence of 16.7%, and the gestational age was 28.7 (27.7, 30.0) weeks, including 454 males (58.4%) and 323 females (41.6%). There were 622 preterm infants in the training cohort, including 105 preterm infants in the PH group. A total of 155 patients were enrolled in the verification cohort, including 25 patients in the PH group. Multivariate Logistic regression analysis revealed that low 5 min Apgar score (OR=0.87, 95%CI 0.76-0.99), cesarean section (OR=1.97, 95%CI 1.13-3.43), small for gestational age (OR=9.30, 95%CI 4.30-20.13), hemodynamically significant patent ductus arteriosus (hsPDA) (OR=4.49, 95%CI 2.58-7.80), late-onset sepsis (LOS) (OR=3.52, 95%CI 1.94-6.38), and ventilator-associated pneumonia (VAP) (OR=8.67, 95%CI 3.98-18.91) were all independent risk factors for PH (all P<0.05). The independent risk factors and the severity of BPD were combined to construct a nomogram map model. The area under the receiver operating characteristic (ROC) curve of the nomogram model in the training cohort and the validation cohort were 0.83 (95%CI 0.79-0.88) and 0.87 (95%CI 0.79-0.95), respectively, and the calibration curve was close to the ideal diagonal. Conclusions: Risk of PH with BPD increases in preterm infants with low 5 minute Apgar score, cesarean section, small for gestational age, hamodynamically significant patent ductus arteriosus, late-onset sepsis, and ventilator-associated pneumonia. This nomogram model serves as a useful tool for predicting the risk of PH with BPD in premature infants, which may facilitate individualized early intervention.

[Myocardial biopsy of Liwen procedure: representability and etiological diagnostic value of cardiac samples obtained by a novel technique in patients with hypertrophic cardiomyopathy].

Objective: To investigate the representability and etiological diagnostic value of myocardium samples obtained from patients with hypertrophic cardiomyopathy (HCM) by transthoracic echocardiography-guided percutaneous intramyocardial septal biopsy (myocardial biopsy of Liwen procedure). Methods: This study was a retrospective case-series analysis. Patients with HCM, who underwent myocardial biopsy of Liwen procedure and radiofrequency ablation in Xijing Hospital, Air Force Military Medical University from July to December 2019, were included. Demographic data (age, sex), echocardiographic data and complications were collected through electronic medical record system. The histological and echocardiographic features, pathological characteristics of the biopsied myocardium of the patients were analyzed. Results: A total of 21 patients (aged (51.2±14.5) years and 13 males (61.9%)) were enrolled. The thickness of ventricular septum was (23.3±4.5)mm and the left ventricular outflow tract gradient was (78.8±42.6)mmHg (1 mmHg=0.133 kPa). Eight patients (38.1%) were complicated with hypertension, 1 patient (4.8%) had diabetes, and 2 patients (9.5%) had atrial fibrillation. Hematoxylin-eosin staining of myocardial samples of HCM patients before radiofrequency ablation evidenced myocytes hypertrophy, myocytes disarray, nuclear hyperchromatism, hypertrophy, atypia, coronary microvessel abnormalities, adipocyte infiltration, inflammatory cell infiltration, cytoplasmic vacuoles, lipofuscin deposition. Interstitial fibrosis and replacement fibrosis were detected in Masson stained biopsy samples. Hematoxylin-eosin staining of myocardial samples of HCM patients after radiofrequency ablation showed significantly reduced myocytes, cracked nuclear in myocytes, coagulative necrosis, border disappearance and nuclear fragmentation. Quantitative analysis of myocardial specimens of HCM patients before radiofrequency ablation showed that there were 9 cases (42.9%) with mild myocardial hypertrophy and 12 cases (57.1%) with severe myocardial hypertrophy. Mild, moderate and severe fibrosis were 5 (23.8%), 9 (42.9%) and 7 (33.3%), respectively. Six cases (28.6%) had myocytes disarray. There were 11 cases (52.4%) of coronary microvessel abnormalities, 4 cases (19.0%) of adipocyte infiltration, 2 cases (9.5%) of inflammatory cell infiltration,6 cases (28.5%) of cytoplasmic vacuole, 16 cases (76.2%) of lipofuscin deposition. The diameter of cardiac myocytes was (25.2±2.8)µm, and the percentage of collagen fiber area was 5.2%(3.0%, 14.6%). One patient had severe replacement fibrosis in the myocardium, with a fibrotic area of 67.0%. The rest of the patients had interstitial fibrosis. The myocardial specimens of 13 patients were examined by transmission electron microscopy. All showed increased myofibrils, and 9 cases had disorder of myofibrils. All patients had irregular shape of myocardial nucleus, partial depression, mild mitochondrial swelling, fracture and reduction of mitochondrial crest, and local aggregation of myofibrillary interfascicles. One patient had hypertrophy of cardiomyocytes, but the arrangement of muscle fibers was roughly normal. There were vacuoles in the cytoplasm, and Periodic acid-Schiff staining was positive. Transmission electron microscopy showed large range of glycogen deposition in the cytoplasm, with occasional double membrane surround, which was highly indicative of glycogen storage disease. No deposition of glycolipid substance in lysozyme was observed under transmission electron microscope in all myocardial specimens, which could basically eliminate Fabry disease. No apple green substance was found under polarized light after Congo red staining, which could basically exclude cardiac amyloidosis. Conclusion: Myocardium biopsied samples obtained by Liwen procedure of HCM patients are representative and helpful for the etiological diagnosis of HCM.

MiR-135b-5p affected malignant behaviors of ovarian cancer cells by targeting KDM5B.

The article "MiR-135b-5p affected malignant behaviors of ovarian cancer cells by targeting KDM5B, by R. Ren, J. Wu, M.-Y. Zhou, published in Eur Rev Med Pharmacol Sci 2020; 24 (7): 3548-3554-DOI: 10.26355/eurrev_202004_20815-PMID: 32329828" has been withdrawn from the authors stating that "some data cannot be repeated by our further research". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20815.

Long noncoding RNA OR3A4 promotes the migration and invasion of melanoma through the PI3K/AKT signaling pathway.

The article "Long noncoding RNA OR3A4 promotes the migration and invasion of melanoma through the PI3K/AKT signaling pathway, by J. Wu, M.-Y. Zhou, X.-P. Yu, Y. Wu, P.-L. Xie, published in Eur Rev Med Pharmacol Sci 2019; 23 (16): 6991-6996-DOI: 10.26355/eurrev_201908_18739-PMID: 31486499" has been withdrawn from the authors. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18739.

[Progress in diagnosis and treatment of rectal neuroendocrine neoplasms].

Neuroendocrine neoplasms (NENs) are relatively rare heterogeneous tumors that originate from peptidergic neurons and neuroendocrine cells and have been referred to as "carcinoids" in the past. Although this type of tumor had been previously considered to be indolent tumor with a low degree of malignancy, with the development of medicine and clinical study, researchers found that NENs had the potential to metastasize. They can occur in any part of the body where neuroendocrine cells are distributed and gastro-entero-pancreatic neuroendocrine neoplasms (GEP-NENs) are the most common type of NENs.Due to the improvement of techniques such as endoscopy and imaging, the incidence of rectal neuroendocrine tumors(R-NENs) and the number of related clinical researches have both increased significantly in recent years. Although researches in Chinese and foreign medical centers are mostly retrospective studies of small samples and the efficacies of different treatment methods are still under debating and lack of sufficient medical evidence to support, the diagnosis and treatment of this disease is gradually becoming standardized according to the proposal of corresponding guidelines. The recent advances in the epidemiology, diagnosis and treatment of rectal neuroendocrine neoplasms are reviewed in this paper.

MiR-135b-5p affected malignant behaviors of ovarian cancer cells by targeting KDM5B.

OBJECTIVE: The aim of this study was to investigate the potential effect of microRNA-135b-5p (miR-135b-5p) on the development of ovarian cancer (OC) and to explore the relevant mechanism. PATIENTS AND METHODS: The expression of miR-135b-5p in OC tissues and cells was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). MicroRNA online prediction websites were used to screen the potential targets of miR-135b-5p. Subsequently, luciferase reporter gene assay and Western blot (WB) were performed for further confirmation. In addition, the effects of miR-135b-5p on cell function were analyzed by relevant experiments in vitro. RESULTS: MiR-135b-5p was lowly expressed in both OC tissues and cell lines. Combined with online prediction software, luciferase reporter gene assay and WB, KDM5B was predicted and verified as a downstream target gene of miR-135b-5p. Down-regulating the expression of KDM5B by over-expressing miR-135b-5p in OC cells could effectively control the proliferation and apoptosis of OC cells. Cell proliferation was significantly reduced, while cell apoptosis was promoted after miR-135b-5p mimics transfection in OC cells. CONCLUSIONS: By targeting KDM5B, miR-135b-5p exerted an excellent anti-cancer effect in OC cells. Our findings indicated that miR-135b-5p/KDM5B might become a feasible and new target of OC treatment.

[Analysis of common gynecological diseases in 1142 married female workers].

Objective: To understand the prevalence of gynecologic diseases among married female workers. Methods: The data of married female workers who underwent occupational health examination in a physical examination center from January to December 2017 were collected. The relationship between the detection of common gynecological diseases, age and occupational types examined by gynecological routine, TCT, breast B-mode ultrasound, uterus and appendix B-mode ultrasound were analyzed. Results: Among the 1142 female workers, the total detection rate of reproductive tract infections was 67.25% (768/1142), the total detection rate of breast-related diseases was 75.22% (859/1142) ; the total detection rate of gynecological tumors and benign lesions was 14.71% (168/1142). The detection rate of breast hyperplasia was the highest 67.08% (766/1142), followed by vaginitis 51.66% (590/1142). Among the abnormalities detected in breast-related diseases, gynecological tumors and benign lesions, the highest detection rate was found in public institutions (85.66% and 27.13%), and the lowest was found in factory workers (70.24% and 7.89%). With the increase of age, the detection rate of breastrelated diseases (breast hyperplasia, breast cyst), gynecological tumors, benign lesions (uterine myoma), and Nessler's cyst abnormalities in married female workers increased (χ(2)(trend)=7.647、21.653、107.411、53.802, P<0.05), while the detection rate of columnar epithelium of cervix decreased (χ(2)(trend)=7.404, P<0.05). There was no significant difference in the total detection rate of reproductive tract infectious diseases (vaginitis, cervical polyps, cervical hypertrophy) among married famale workers of different ages (P<0.05) . Conclusion: The common gynecological diseases of married female workers are affected by many factors such as age and occupation. Health examination and health education should be carried out regularly to reduce the incidence of gynecological diseases among female workers according to different ages and occupations.

Long noncoding RNA OR3A4 promotes the migration and invasion of melanoma through the PI3K/AKT signaling pathway.

OBJECTIVE: Recent studies have revealed the crucial role of long non-coding RNAs (lncRNAs) in tumor progression. This study aims to identify the biological function of lncRNA OR3A4 in the progression of melanoma. PATIENTS AND METHODS: OR3A4 expression in melanoma cells and tissue samples was detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). The regulatory effects of OR3A4 on melanoma cells were identified by performing transwell assay and wound healing assay in vitro. The underlying mechanism of OR3A4 in mediating the progression of melanoma was explored by RT-qPCR and Western blot. RESULTS: OR3A4 expression was remarkably upregulated in melanoma tissues compared with normal tissues. Moreover, migration and invasion of melanoma cells were inhibited after knockdown of OR3A4 in vitro, which were promoted after overexpression of OR3A4. Furthermore, the targeted proteins in phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway were downregulated after knockdown of OR3A4 in vitro, which were upregulated by overexpressed OR3A4. CONCLUSIONS: OR3A4 could promote the invasion and migration of melanoma cells by inducing the PI3K/AKT signaling pathway, which may offer a new therapeutic intervention for melanoma patients.

[The inhibition effects of apatinib on cell proliferation, migration and apoptosis in esophageal carcinoma via Ras/Raf/MEK/ERK and JAK2/STAT3 pathways].

Objective: To investigate the in vitro and in vivo effects of apatinib in esophageal squamous cell carcinoma and the underlying mechanisms. Methods: The esophageal cancer cells, KYSE-150 and ECA-109, were divided into control group and apatinib treatment group at the concentrations of 2.5, 5, 10, 20 and 40 µmol/L respectively. All of experiments were performed in triplicate. MTT and colony formation assays were used to measure cell proliferation. Transwell assay was used to determine the migration capacity. The effect of apatinib on cell cycle and apoptosis was analyzed by flow cytometry. The expression of VEGF and VEGFR-2 was measured by real-time quantitative PCR (qRT-PCR). The concentration of VEGF in the cell supernatant was assessed by enzyme-linked immunosorbent assay (ELISA). The expression levels of MEK, ERK, p-MEK, p-ERK, JAK2, STAT3 and p-STAT3 after VEGF stimulation were detected by Western blot. Furthermore, the nude mice xenograft model was established. The tumor-bearing mice were randomly divided into control group, apatinib low dose treatment group (250 mg) and apatinib high dose treatment group (500 mg), respectively. Tumor inhibition rates of different groups were calculated. And then the expressions of VEGF and VEGFR2 were detected in xenograft tissues by immunohistochemical staining. Results: In the presence of 20 µmol/L and 40 µmol/L of apatinib for 24 hours, the migration cell numbers of KYSE-150 and ECA-109 were 428.67±4.16 and 286.67±1.53 as well as 1 123.67±70.00 and 477.33±26.84, respectively, that were significantly lower than control group (P<0.05 for all). In addition, after treatment with 10 µmol/L, 20 µmol/L and 40 µmol/L of apatinib for 7 days on KYSE-150 and ECA-109, the colony formation rates were (65.12±25.48)%, (58.19±24.73)% and (29.10±22.40)% as well as (70.61±15.14)%, (61.12±17.21)% and (43.09±11.13)%, respectively. The colony formation rates of 20 µmol/L and 40 µmol/L of apatinib treatment groups were significantly lower than control group (100.00±0.00, P<0.05). The cell cycle ratio of G(2)/M phase and apoptosis rate of control group and 20 µmol/L apatinib group in KYSE-150 cells were (12.14±2.13)% and (3.49±0.74)% as well as (26.27±3.30)% and (15.65±1.54)%, respectively. The corresponding ratios in ECA-109 cells were (3.44±0.57)% and (6.31±1.43)% as well as (22.64±2.36)% and (49.26±1.62)%, respectively. The results show that apatinib suppressed cell cycle progression at G(2)/M phase and induced cell apoptosis in both KYSE-150 and ECA-109 cells (P<0.05 for all). In the presence of 20 µmol/L and 40 µmol/L of apatinib in KYSE-150 cells, the relative levels of VEGF mRNA were (42.57±10.43)% and (25.69±1.24)%, and those of VEGF-2 mRNA were (36.09±10.82)% and (13.99±6.54)%, which were all significantly decreased compared to control group (100.00±0.00, P<0.05 for all). For ECA-109 cells, the relative expression of VEGF and VEGFR2 showed similar tendency (P<0.05 for all). Moreover, after treatment with 20 µmol/L and 40 µmol/L of apatinib in KYSE-150 cells, the VEGF concentrations were (766.48±114.27) pg/ml and (497.40±102.18)pg/ml, which were significantly decreased compared to control group [(967.41±57.75) pg/ml, P<0.05)]. The results in ECA-109 were consistent (P<0.05). Furthermore, after treatment with 40 µmol/L of apatinib in KYSE-150 and ECA-109, the relative expression of p-MEK and p-ERK were 0.49±0.05 and 0.28±0.03 as well as 0.63±0.03 and 1.22±0.15, which were significantly lower than control group (1.23±0.19 and 0.66±0.07 as well as 1.03±0.20 and 1.76±0.20; P<0.05). The relative expression of STAT3, p-STAT3 in control group and experimental group were 0.96±0.15 and 0.85±0.16 as well as 0.62±0.09 and 0.36±0.13, respectively. The results showed that the protein levels of STAT3 and p-STAT3 were significantly lower than the control group (P<0.05 for all). The inhibition rates of apatinib in xenograft nude mice were 29.25% and 19.96% for 250 mg and 500 mg treatment groups. The concentration of VEGF were (25.11±4.12) pg/ml, (16.40±2.81) pg/ml and (15.04±4.88)pg/ml for control, 250 mg and 500 mg treatment groups, respectively. Conclusions: Apatinib can inhibit cell proliferation, induce apoptosis and suppress migration of esophageal cancer cells in vitro and in vivo. This effect was mainly mediated via the alterations of Ras/Raf/MEK/ERK pathway and JAK2/STAT3 pathway.

Effect of excess dietary fluoride on laying performance and antioxidant capacity of laying hens.

The objective of this study was to investigate the effects of excess dietary fluoride (F) on laying performance and antioxidant capacity of laying hens. A total of 576 laying hens, 51 wk old, was randomly divided into 6 groups, each of which included 6 replicates of 16 hens. Graded amounts of sodium fluoride (NaF) were added to the basal diet to achieve concentrations of 16 (control), 200, 400, 600, 800, and 1,000 mg/kg F, respectively. Dietary F at 1,000 mg/kg significantly decreased ADFI, laying rate, and average egg weight, and increased feed conversion ratio (FCR) (P < 0.05). No significant differences were observed in serum total antioxidant capacity (T-AOC) level or catalase (CAT) concentration among all the treatments, while hens fed F at 800 and 1,000 mg/kg had higher activity of serum glutathione peroxidase (GSH-PX) and concentration of malondialdehyde (MDA) (P < 0.05) as compared to the control group. Compared with the control group, dietary F at 400 mg/kg increased liver MDA concentration (P < 0.001), and decreased CAT concentration of liver (P < 0.001); 600 mg/kg F decreased liver T-AOC levels (P < 0.001); and 800 mg/kg of F decreased liver total superoxide dismutases (T-SOD) activity (P < 0.001). Compared with the control group, feeding F at 600 mg/kg decreased kidney T-AOC levels and T-SOD activity (P < 0.001), and increased MDA concentration of kidney (P < 0.001), while dietary 1,000 mg/kg of F decreased kidney GSH-PX activity (P < 0.05) and CAT concentration (P < 0.001). In conclusion, these results indicated that excessive F ingestion had an adverse effect on laying performance by inducing oxidative stress and impairing the antioxidant system of laying hens.

[Analysis of sonographic characteristics of thyroid nodules diagnosed as false positive or negative results in shear wave elastography].

Objective:In order to improve diagnostic accuracy, we study the characteristics of two dimensional ultrasound and shear wave elastography in the diagnosis of false negative or false positive thyroid nodules by shear wave elastography.Method:One hundred and eighty-nine nodules in 189 consecutive patients who had been determined by surgical operation and pathology. Conventional ultrasound features and SWE elasticity imaging characteristics and properties of the final postoperative pathology were recorded. A comparative study between true and false results of quantitative SWE elasticity imaging, and the corresponding conventional ultrasound nodule characteristics were compared.Result:Postoperative pathology showed 189 nodules, 74(39.2%) were benign and 115(60.8%) were malignant. The sensitivity, specificity of conventional ultrasound in the diagnosis of thyroid nodules were 56.5% and 81.1% respectively, and those of SWE were 60.9% and 85.1%. The false positive rate of shear wave elastography in diagnosing benign nodules and the false negative rate of malignant nodules were 14.9% and 39.1%, respectively. The false negative rate was higher than the false positive rate. A vertical growth (P< 0.01) and smaller diameter of the masses were significantly associated with false SWE findings (P< 0.01).Conclusion:The SWE imaging has important significance for differentiating benign and malignant thyroid nodules, but false results are inevitable, which requires clinicians conjunction with other test results to prevent errors judgment when reviewing the SWE imaging.

Effects of arsenic supplementation in feed on laying performance, arsenic retention of eggs and organs, biochemical indices and endocrine hormones.

1. The primary objective of this experiment was to estimate the toxic effects of arsenic (As) supplementation in feed on laying performance, As retention by eggs and organs, serum biochemical indices and endocrine hormones in laying hens. 2. A total of 320 "Jinghong Number 1" hens, 56-week-old, were randomly allocated into four treatments of four replicates with 20 layers in each. Graded arsenical was added to the basal diet in the experimental diets at As levels of 0, 17, 34 and 51 mg/kg, respectively. The trial lasted for 9 weeks including 1 week for acclimatisation. 3. Supplementation of dietary As for eight weeks had no effect on laying performance. As retention in albumen, yolk, egg, liver and kidney increased as As levels increased The level of serum phosphorus (P) was minimised at the 17 mg As/kg group. The activity of serum glutamic oxaloacetic transaminase (GOT) increased linearly. No differences were observed for levels of serum calcium (Ca), alkaline phosphatase (AKP) and serum glutamic pyruvic transaminase (GPT). Concentrations of estradiol (E2) and progesterone (PG) declined at 34 and 51 mg/kg As levels compared with the control group. As supplementation exerted no influence on levels of serum follicle stimulating hormone (FSH), luteinising hormone (LH), triiodothyronine (T3), thyroxine (T4) and the ratio between T3 and T4. 4. In conclusion, dietary As supplementation accelerated retention in tissues and eggs, and affected the laying rate by diminishing hormone levels of E2 and PG at 51 mg/kg.

Physical and functional interaction of Runt-related protein 1 with hypoxia-inducible factor-1alpha.

Angiogenesis and hematopoiesis are closely linked and interactive with each other, but few studies were given to identify possible links between angiogenesis-promoting proteins and hematopoiesis-related transcription factors. Here we investigated the potential relationship of oxygen-sensitive alpha-subunit of angiogenesis-related hypoxia-inducible factor-1alpha (HIF-1alpha) with Runt-related protein 1 (Runx1, also known as acute myeloid leukemia-1, AML-1), an important hematopoietic transcription factor. The results demonstrated that Runx1 and HIF-1alpha proteins directly interacted with each other to a degree, in which Runt homology domain of Runx1 was mainly involved. Leukemia-related abnormal Runx1 fusion protein AML1-ETO, which fuses the N-terminal 177 amino acid residues of the Runx1 protein in frame to ETO (eight-twenty-one) protein, also interacted with HIF-1alpha protein with greater ability than Runx1 itself. More intriguingly, Runx1 overexpression inhibited DNA-binding and transcriptional activity of HIF-1 protein with reduced expression of HIF-1-targeted genes such as vascular endothelial growth factor, while silence of Runx1 expression by specific small interfering RNA significantly increased transcriptional activity of HIF-1 protein, suggesting that Runx1 inhibited transcription-dependent function of HIF-1. Vice versa, HIF-1alpha increased DNA-binding ability and transcriptional activity of Runx1 protein. All these data would shed new insight to understanding Runx1 and HIF-1alpha-related hematopoietic cell differentiation and angiogenesis.

CCAAT/enhancer-binding protein alpha antagonizes transcriptional activity of hypoxia-inducible factor 1 alpha with direct protein-protein interaction.

Hypoxia-inducible factor 1 (HIF-1), a master heterodimeric transcriptional regulator consisting of HIF-1alpha and HIF-1beta subunits for cellular response to hypoxia, plays an important role in carcinogenesis, while CCAAT/enhancer-binding protein alpha (C/EBPalpha) is proposed to act as a tumor suppressor in C/EBPalpha-expressing tissues. Previously, we reported that ectopically expressed HIF-1alpha protein interacts with and enhances transcriptional activity of C/EBPalpha, which favors leukemic cell differentiation. Here we further showed that such an interaction also occurred in their endogenously expressing state of leukemic U937 cells. Glutathione S-transferase pull-down assay proposed that the protein-protein interaction was direct, and transactivation domains of C/EBPalpha and the basic helix-loop-helix domain of HIF-1alpha were essential for such an interaction. More intriguingly, we provided the first demonstration that C/EBPalpha competed with HIF-1beta for direct binding to HIF-1alpha protein. Correspondingly, C/EBPalpha overexpression significantly inhibited the DNA-binding ability of HIF-1 and expressions of hypoxia-responsive element-driven luciferase and HIF-1-targeted genes vascular endothelial growth factor, glucose transporter-1 and phosphoglycerate kinase 1. In parallel, suppression of C/EBPalpha expression by specific small hairpin RNA increased DNA-binding ability of HIF-1 and expression of these HIF-1-targeted genes in leukemic U937 cells. These results would provide new insights for antitumor potential of C/EBPalpha protein.

HLA-G expression in human ovarian carcinoma counteracts NK cell function.

BACKGROUND: Human leukocyte antigen-G (HLA-G) is an important immunotolerant which could be a part of the strategies applied by malignant cells applied to avoid host immunosurveillance. Aberrant expression of HLA-G has been found in ovarian carcinoma. The aim of this study was to evaluate the HLA-G expression in ovarian cancer tissues and to explore its function in vitro. MATERIALS AND METHODS: HLA-G expression in 33 primary ovarian carcinoma tissues was analyzed using immunohistochemistry with the anti-HLA-G monoclonal antibody (mAb) 4H84. Furthermore, the function of HLA-G in NK cell cytotoxicity was determined in vitro by cloning and expression of HLA-G on the ovarian carcinoma cell OVCAR-3. RESULTS: HLA-G expression was detected in 22/33 (66.7%) primary tumor tissues, but was absent in normal ovarian tissues (P<0.01). Cytotoxicity studies showed that HLA-G expression dramatically inhibits cell lyses by NK-92 cells (P<0.01), which could be restored by the anti-HLA-G conformational mAb 87G (P<0.01). CONCLUSION: HLA-G was expressed in a significant number of primary ovarian carcinoma tissues, and HLA-G expression in OVCAR-3 could directly inhibit NK-92 cell lysis. Taken together, our results indicated that expression of HLA-G plays an important role in evasion of ovarian cancer cells from host immunosurveillance.

Angiotensin stimulates the expression of interferon-inducible genes in H295R cells.

Angiotensin-II (A-II) induces proliferation of zona glomerulosa cells and stimulates expression of cytochrome P-450 aldosterone synthase. The genes activated during this adrenal remodeling are not well defined. To clarify this mechanism, we sought to identify the genes whose expression is stimulated by A-II in the H295R cell line. Using a subtractive hybridization technique, we identified one clone whose expression was stimulated by A-II. The sequence of this gene was homologous to the human interferon-inducible genes, 9-27, 1-8D and 1-8U. The 5' portion of the gene was identical to the 1-8D gene product and the 3' was identical to the 9-27 gene product, but the existence of a transcript was not demonstrated by RT-PCR. The expression of these three genes was stimulated by A-II, with the 9-27 gene being most abundant. Potassium and forskolin also stimulated the expression of the 9-27 gene in the H295R cells, but not as effectively as did A-II or interferon-gamma.

[Expression of EGFR and PCNA, and DNA content in squamous cell carcinoma of larynx].

OBJECTIVE: To investigate the expression of epidermal growth factor (EGF), proliferating cell nuclear antigen (PCNA) and DNA index (DI) in laryngeal carcinoma, to analyse the correlation between these index and the biological characteristics of laryngeal carcinoma and their values of clinical prognosis. METHOD: Immunohistochemical staining was used to detect the expression of EGFR and PCNA in laryngeal cancer and normal tissue, and with MIPS-I image analysis system DNA contents of cancer cell were measured and made out DNA index. RESULT: The positive rate of EGFR in laryngeal carcinoma was 54.8%, and it was negative in all 10 normal laryngeal mucosa specimens (P < 0.01). The expression of EGFR did not correlate with histological grading and 5-years survival rate (P > 0.05), The positive expression of PCNA and DNA contents in the laryngeal carcinoma were increased with the decrease of tumorous differentiation (P < 0.05). With the increasing of PCNA positive expression and DI, the prognosis of the patients were poorer (P < 0.05). CONCLUSION: EGFR may be related to the process of carcinogenesis in laryngeal carcinoma and was used as an early biomarker identifying premalignant lesions which had the greatest risk of carcinogenesis. PCNA and DI were simultaneously detected can be used as the prediction of tumor malignancy and prognosis.

Universal TA cloning.

TA cloning is one of the simplest and most efficient methods for the cloning of PCR products. The procedure exploits the terminal transferase activity of certain thermophilic DNA polymerases, including Thermus aquaticus (Taq) polymerase. Taq polymerase has non-template dependent activity which preferentially adds a single adenosine to the 3'-ends of a double stranded DNA molecule, and thus most of the molecules PCR amplified by Taq polymerase possess single 3'-A overhangs. The use of a linearized "T-vector" which has single 3'-T overhangs on both ends allows direct, high-efficiency cloning of PCR products, facilitated by complementarity between the PCR product 3'-A overhangs and vector 3'-T overhangs. The TA cloning method can be easily modified so that the same T-vector can be used to clone any double-stranded DNA fragment, including PCR products amplified by any DNA polymerase, as well as all blunt- and sticky-ended DNA species. This technique is especially useful when compatible restriction sites are not available for the subcloning of DNA fragments from one vector to another. Directional cloning is made possible by appropriate hemi-phosphorylation of both the T-vectors and the inserts. With a single T-vector at hand, any DNA fragment can be cloned without compromising the cloning efficiency. The universal TA cloning method is thus both convenient and labor-saving.