Bullying victimization and internalizing and externalizing problems in school-aged children: The mediating role of sleep disturbance and the moderating role of parental attachment.

BACKGROUND: Studies have shown that bullying victimization may be related to internalizing and externalizing problems; however, the mechanism underlying this relationship remains unknown. This study explored the mediating role of sleep disturbance and the moderating role of parental attachment. METHODS: A total of 1543 Chinese primary school students (M age = 8.92 years, SD1.7 years; range, 6-12) completed bullying victimization, sleep disturbance, and parental attachment measures, and provided information on their parents' occupations. The parents or guardians (n = 1995) also completed ratings on their children's internalizing and externalizing problems. RESULTS: It was found that bullying victimization directly affected internalizing and externalizing problems and also influenced sleep disturbance. Regardless of the parent's socioeconomic status, parental attachment was found to moderate the relationship between bullying victimization and internalizing problems. CONCLUSIONS: These findings contribute to understanding the partial mediating mechanism of sleep disturbance in the association between bullying victimization and internalizing and externalizing problems. The protective role of parental attachment proved central to preventing internalizing problems in bullied children. Intervention programs that enhance parental attachment and improve sleep quality could assist in mitigating the impact of bullying victimization on internalizing or externalizing problems.

Exploring the mechanism of Buxue Yimu Pill on hemorrhagic anemia through molecular docking, network pharmacology and experimental validation.

Buxue Yimu Pill (BYP) is a classic gynecological medicine in China, which is composed of Angelica sinensis (Oliv.) Diels, Leonurus japonicus Houtt, Astragalus membranaceus (Fisch.) Bunge, Colla corii asini and Citrus reticulata Blanco. It has been widely used in clinical therapy with the function of enriching Blood, nourishing Qi, and removing blood stasis. The current study was designed to determine the bioactive molecules and therapeutic mechanism of BYP against hemorrhagic anemia. Herein, GC-MS and UPLC/Q-TOF-MS/MS were employed to identify the chemical compounds from BYP. The genecards database (https: //www.genecards.org/) was used to obtain the potential target proteins related to hemorrhagic anemia. Autodock/Vina was adopted to evaluate the binding ability of protein receptors and chemical ligands. Gene ontology and KEGG pathway enrichment analysis were conducted using the ClusterProfiler. As a result, a total of 62 candidate molecules were identified and 152 targets related to hemorrhagic anemia were obtained. Furthermore, 34 active molecules and 140 targets were obtained through the virtual screening experiment. The data of molecular-target (M-T), target-pathway (T-P), and molecular-target-pathway (M-T-P) network suggested that 32 active molecules enhanced hematopoiesis and activated the immune system by regulating 57 important targets. Pharmacological experiments showed that BYP significantly increased the counts of RBC, HGB, and HCT, and significantly down-regulated the expression of EPO, IL-6, CSF3, NOS2, VEGFA, PDGFRB, and TGFB1. The results also showed that leonurine, leonuriside B, leosibiricin, ononin, rutin, astragaloside I, riligustilide and levistolide A, were the active molecules closely related to enriching Blood. In conclusion, based on molecular docking, network pharmacology and validation experiment results, the enriching blood effect of BYP on hemorrhagic anemia may be associated with hematopoiesis, anti-inflammation, and immunity enhancement.

Inhibitory Role of an Aeromonas hydrophila TIR Domain Effector in Antibacterial Immunity by Targeting TLR Signaling Complexes in Zebrafish.

The Toll/interleukin-1 receptor (TIR) domain is a structural unit responsible for the assembly of signal protein complexes in Toll-like receptor (TLR) and interleukin-1 receptor signaling pathways. TIR domain homologs are found in a considerable number of bacteria and enhance bacterial infection and survival in host organisms. However, whether TIR domain homologs exist in Aeromonas hydrophila, a ubiquitous waterborne bacterium in aquatic environments, remains poorly understood. In this study, a TIR domain protein (TcpAh) was identified from A. hydrophila JBN2301. TIR domain of TcpAh is highly homologous to the counterpart domains in TLRs and myeloid differentiation factor 88 (MyD88). The zebrafish infected with mutant A. hydrophila with tcpAh deletion had a remarkably lower mortality than those infected with the wild-type strain. This result suggests that TcpAh is a crucial virulence factor for A. hydrophila infection. TcpAh exhibited a strong ability to associate with MyD88, tumor necrosis factor receptor-associated factor 3 (TRAF3) and TRAF-associated NF-κB activator-binding kinase 1 (TBK1) in TIR-TIR, TIR-Death domain (DD), and other alternative interactions. This finding suggests that TcpAh extensively interferes with MyD88 and TIR domain-containing adapter inducing interferon (IFN)-ß (TRIF) signaling pathways downstream of TLRs. Consequently, CD80/86 expression was suppressed by TcpAh via attenuating TLR-stimulated NF-κB activation, which ultimately led to the impairment of the major costimulatory signal essential for the initiation of adaptive humoral immunity against A. hydrophila infection. We believe that this study is the first to show a previously unrecognized mechanism underlying A. hydrophila evades from host antibacterial defense by intervening CD80/86 signal, which bridges innate and adaptive immunity. The mechanism will benefit the development of therapeutic interventions for A. hydrophila infection and septicemia by targeting TcpAh homologs.

Associations between hepatitis B virus exposure and the risk of extrahepatic digestive system cancers: A hospital-based, case-control study (SIGES).

OBJECTIVES: This case-control study was aimed to investigate associations between HBV infection and extrahepatic digestive system cancers. METHODS: The patients of gastric, small intestinal, colonic, rectal, anal, biliary tract, and pancreatic cancers were retrospectively collected between 2016.5 and 2017.12. Simultaneously, the healthy controls were collected from the health check-up registry, and cancer-free status was confirmed based on medical records. Propensity score matching was performed to reduce bias. Multinomial logit model and conditional logistic regression model were used to assess the risk of individual cancer according to HBV serological markers and classifications. RESULTS: Totally, 4748 patients involving seven cancers, and 57,499 controls were included. After matching, HBsAg was associated with increased risk of gastric cancer (aOR = 1.39, 95% CI: 1.05-1.85), and anti-HBs served as a protective factor for gastric (aOR = 0.72, 95% CI: 0.61-0.85), colonic (aOR = 0.73, 95% CI: 0.60-0.89), rectal (aOR = 0.73, 95% CI: 0.63-0.85), and pancreatic (aOR = 0.58, 95% CI: 0.42-0.82) cancers. Compared to subgroups with non-infection and vaccination status, inactive HBsAg carriers and active HBV infection subgroup were correlated with gastric carcinogenesis (aOR = 1.41, 95% CI: 1.03-1.93). However, no clear association was found between HBV infection and other cancers. CONCLUSIONS: HBV infection was potentially associated with an increased risk of gastric cancer. The development mechanism of HBV-associated gastric cancer needs to investigate further.

Predictors of pregnancy after intrauterine insemination in women with polycystic ovary syndrome.

OBJECTIVE: To evaluate the effects of body mass index (BMI) in patients with polycystic ovary syndrome (PCOS) undergoing controlled ovarian stimulation (COS) with intrauterine insemination (IUI). METHODS: This retrospective study evaluated couples with PCOS undergoing COS and IUI. The relationship between cumulative IUI pregnancy outcomes and BMI, treatment cycles, treatment schemes, number of dominant follicles, endometrial thickness, infertility duration and type of infertility was analysed. RESULTS: The study evaluated 831 IUI cycles in 451 couples with PCOS. Compared with normoweight women, overweight and obese women required more human menopausal gonadotropin (hMG) doses and more days of COS. Gestational diabetes mellitus occurred more frequently in the obese group than in the other BMI groups. The clinical pregnancy and live birth rates in the hMG, clomiphene citrate (CC) + hMG and letrozole (LE) + hMG groups were significantly higher than those in the CC and LE groups. The clinical pregnancy rate was higher in the secondary infertility group compared with the primary infertility group. CONCLUSION: Obese women might require more hMG doses and more days of COS to overcome the effects of weight. As BMI increases, the incidence of gestational diabetes might also increase. The number of cycles and type of infertility may have a predictive value for pregnancy outcomes.

Gamma-glutamyl transpeptidase-to-platelet ratio and the fibrosis-4 index in predicting hepatitis B virus-related hepatocellular carcinoma development in elderly chronic hepatitis B patients in China: A single-center retrospective study.

Gamma-glutamyl transpeptidase-to-platelet ratio (GPR) and fibrosis-4 (FIB-4) index have been reported to be useful predictors in predicting hepatocellular carcinoma (HCC) development in chronic hepatitis B (CHB) patients. However, their predictive performances on HCC development have not been validated in elderly patients. Thus, the aim of this study was to evaluate the predictive values of the GPR and FIB-4 index on HCC in elderly CHB patients with in China.Between January 2007 and December 2016, 1011 CHB patients older than 60 years were enrolled in the study, and their data were retrospectively analyzed. Receiver-operating characteristic (ROC) curve analysis was used to determine the optimal cutoff points of GPR and the FIB-4 index. Cumulative HCC incidence rates were calculated by the Kaplan-Meier method and compared by the log-rank test. Univariate and multivariate analyses were performed to detect risk factors for HCC development. The prediction performances of GPR and FIB-4 index were compared based on time-dependent ROC analyses.After a median follow-up of 6.8 (interquartile range 3.9-8.4) years, 39 (3.9%) patients developed HCC. The ROC analysis of GPR and the FIB-4 index at the 5-year time point revealed that the optimal cutoff point was 0.23 for GPR and 4.15 for the FIB-4 index. When stratified by low and high GPR values and FIB-4 indices, the patients' subgroups showed significantly different cumulative incidences of HCC. The multivariate analysis revealed that high GPR (hazard ratio [HR] 4.224; 95% confidence interval [CI] 1.891-9.434, P < .001) was an independent risk factor for HCC development, whereas a high FIB-4 index was not (HR 0.470; 95% CI 0.212-1.043; P = .063). In the time-dependent ROC analysis, GPR showed higher area under curve (AUC) values than the FIB-4 index did at all time points and reached statistical significance at the 5-, 7-, and 10-year time points (GPR vs FIB-4 index, AUC 0.725 vs 0.549 at 5 years, P = .005; GPR vs FIB-4 index, AUC 0.733 vs 0.578 at 7 years, P = .001; GPR vs FIB-4 index, AUC 0.837 vs 0.475 at 10 years, P < .001).In conclusion, our study suggests GPR is superior to the FIB-4 index in predicting HCC development in elderly CHB patients in China.

The risk of stroke and associated risk factors in a health examination population: A cross-sectional study.

Health examination is an important method for early detection of people with different risk of stroke. This study estimates the risk of stroke and identify risk factors for people who underwent health examinations at the Health Examination Center at West China Hospital, Sichuan University from July 2014 to February 2018.A total of 31,464 people were recruited in this study and divided into 3 groups (low risk, moderate risk, and high risk) according to risk of stroke. We explored possible factors associated with the risk of stroke by using multivariable stepwise logistic regression analysis.Among the participants, 17,959 were at low risk, 11,825 were at moderate risk, and 1680 were at high risk. Age, smoking, alcohol consumption, body mass index, uric acid, diastolic pressure, systolic pressure, triglycerides, low-density lipoprotein cholesterol, glucose, and brachial-ankle pulse wave velocity (baPWV) were independent significant risk factors for stroke, whereas high-density lipoprotein cholesterol was an independent protective factor for stroke. Interestingly, with increasing age, the percentage of people at moderate or high risk of stroke was increased. The percentages of people at moderate and high risk of stroke were also increased with respect to the stages of baPWV.This study showed that >40% of the participants were at moderate or high risk of stroke, especially the older participants. Several factors were related to the risk of stroke, especially baPWV. Some preventive action may be adopted early, and more attention can be paid to the health examination population.

Multiple sclerosis and stroke: a systematic review and meta-analysis.

BACKGROUND: Multiple sclerosis (MS) and stroke are two common causes of death and disability worldwide. The relationship between these two diseases remains unclear. Effective early preventative measures and treatments are available to reduce the morbidity and mortality of acute stroke. The objectives of our systematic review are to estimate the risk of stroke in patients with MS and to collate related studies to draw preliminary conclusions that may improve clinical practice. METHOD: Relevant studies were systematically searched in MEDLINE, Embase, the Chinese Biomedical Literature Database (CBM), the China National Knowledge Infrastructure and the VIP database of Chinese periodicals from January 1983 to May 2017, with no restrictions on language. Patients included in this review were adults who suffered from MS. Review Manager 5.3 software program was used to pool data and calculate the risk ratio (RR) and its 95% confidence interval (CI). We also performed heterogeneity and sensitivity analyses and evaluated bias in the meta-analysis. RESULTS: Nine studies including more than 380,000 participants that met our inclusion criteria were incorporated into the meta-analysis. During different follow-up periods, patients with MS had an increased risk of any type of stroke [RR = 3.48, 95% CI (1.59, 7.64), P = 0.002 for 1 year; RR = 2.45, 95% CI (1.90, 3.16), P < 0.00001 for 10-13 years]. The total prevalence of stroke (any type) in patients with MS exceeded expectations compared to different groups [Comparing with general veteran: RR = 2, 95% CI (1.19, 3.38), P = 0.009. Comparing with general population: RR = 2.93, 95% CI (1.13, 7.62), P = 0.03]. Furthermore, ischemic stroke was particularly more common in the MS population than in people without MS [RR = 6.09, 95% CI (3.44, 10.77), P < 0.00001]. CONCLUSION: Compared with the general population, people with MS have an increased risk of developing any type of stroke and ischemic stroke in particular. Consistent results were obtained from patients of different sexes and age groups. Preventative measures and treatments should be administered at earlier time points to improve patient outcomes.

Long noncoding RNA LINC00483/microRNA-144 regulates radiosensitivity and epithelial-mesenchymal transition in lung adenocarcinoma by interacting with HOXA10.

Lung adenocarcinoma (LAD) is the leading cause of cancer death worldwide. Long noncoding RNAs (lncRNAs) have been shown to play an important regulatory role in cancer biology, including that of LAD. The aim of this experiment was to explore the interaction of LINC00483, microRNA-144 (miR-144), and homeobox A10 (HOXA10), and their effects on radio sensitivity and epithelial-mesenchymal transition (EMT) of LAD. Initially, microarray analysis was used to screen out miRNAs and lncRNAs, as well as the differentially expressed genes related to LAD. Following the screening process, the targeting relationship of LINC00483, miR-144, and that of miR-144 and HOXA10 was determined. Following that, the expression of LINC00483, miR-144, messenger RNA (mRNA), as well as protein expression of HOXA10, MMP-2, MMP-9, E-cadherin, vimentin, and N-cadherin that followed in cells was determined. Also, the effect of LINC00483 on cell migration and invasion ability, and cell tumorigenic ability was detected. LINC00483 and HOXA10 were found to be upregulated whereas miR-144 was downregulated in LAD. Silencing of LINC00483 could competitively bind to miR-144, thereby upregulating HOXA10. LINC00483 or HOXA10 silencing led to decreased HOXA10, MMP-2, MMP-9, vimentin, and N-cadherin but elevated miR-144 and E-cadherin. Moreover, after being transfected with silenced LINC00483, the cell proliferation, migration, and invasion were inhibited with enhanced radiosensitivity. Consequently, the data of the study indicates that interference of LINC00483 weakens its competitive binding ability to miR-144, thus reducing HOXA10 expression, and enhancing radiosensitivity in LAD.

[Component Changes of Metabolic Syndrome in Pre-elderly People with Healthy Obese Phenotype].

OBJECTIVE: To determine component changes of metabolic syndrome in pre-elderly people with healthy obese phenotype. METHODS: A total of 1 686 adults aged between 45-59 yr. who underwent health examinations from 2010 to 2016 in West China Hospital of Sichuan University participated in this study. The participants had healthy obese phenotype at the baseline but no history of diabetes,high blood pressure,high cholesterol and cardiovascular disease. Component changes of metabolic syndrome (MS) and associated factors over the seven-year period were analysed using logistic regression modeling. RESULTS: The number of MS components increased over the years in centrally obese individuals,and 11.0% developed MS,including 118 men [(53.29±4.00) years old,66.95% current smokers,5.93% past smokers,24.58% alcohol drinkers] and 67 women [(52.01±4.06) years old,26.87% current smokers,1.49% past smokers,11.94% alcohol drinkers]. The most frequently presented MS components included higher fasting glucose,higher blood pressure and higher triglyceride. Healthy status (0 MS component) resumed in 44 participants who had abdominal obesity (1 MS component) at the baseline: 27 women and 17 men. Age (OR=1.732, 95%CI：1.594-1.882, P<0.000 1),smoking (OR=7.188, 95%CI：4.311-11.986, P<0.000 1) and drinking (OR=3.986, 95%CI：2.283-6.959, P<0.000 1) were identified as risk factors of MS. CONCLUSION: MS components increase over years in both men and women. Smoking and drinking are the main risk factors of MS progression. Regular MS surveillance and behavioral interventions are recommended for pre-elderly people with healthy obese phenotype.

[Clinical Research on the Post-operative Hemodynamics Changes in Donors after Living Donor Liver Transplantation].

OBJECTIVE: To investigate the postoperative hemodynamics changes and their influence factors in the donors after right lobe living donor liver transplantation. METHODS: A total of 53 consecutive living donors from Dec 2010 to Aug 2015 who underwent donor right lobe hepatectomy were retrospectively analyzed. We measured residual liver hemodynamics with color doppler ultrasound, detected liver stiffness by transient elastography, also analyzed postoperative liver function, hemodynamics, and the long term variation tendency of hepatocirrhosis and spleen. RESULTS: One week after operation, transient liver damage was observed. Post-operative hemodynamics within the follow-up time showed:portal vein diameter was gradually increasing, the velocity decreased gradually;Hepatic vein diameter increased, and the velocity decreased gradually. There was a negative correlation between portal vein diameter and portal vein velocity (P=0.012, r=-3.11). Liver stiffness (Kpa value) decreased gradually with time, while spleen volume gradually increased. Correlation analysis showed that postoperative liver stiffness (Kpa value) was negatively related to portal vein diameter (P=0.013, r=-0.338) and positively related to hepatic venous velocity (P=0.038,r=0.246). CONCLUSIONS: The donor presented a transient liver injury after operation, but tended to be recovery after one week. Despite post-operative hemodynamic undergo a series of changes, but it does not affect the post-operative long-term donor safety.

The Ethanol Extract of Licorice (Glycyrrhiza uralensis) Protects against Triptolide-Induced Oxidative Stress through Activation of Nrf2.

To investigate the potential role of nuclear factor erythroid 2-related factor 2 (Nrf2) in licorice ethanol extract (LEE) against triptolide- (TP-) induced hepatotoxicity, HepG2 cells were exposed to LEE (30, 60, and 90 mg·L-1) for 12 h and then treated with TP (50 nM) for 24 h. Besides, an acute liver injury model was established in ICR mice by a single dose of TP (1.0 mg·kg-1, i.p.). Relevant oxidant and antioxidant mediators were analyzed. TP led to an obvious oxidative stress as evidenced by increasing levels of ROS and decreasing GSH contents in HepG2 cells. In vitro results were likely to hold true in in vivo experiments. LEE protected against TP-induced oxidative stress in both in vitro and in vivo conditions. Furthermore, the decreased level of Nrf2 in the TP-treated group was observed. The mRNA levels of downstream genes decreased as well in ICR mice liver, whereas they increased in HepG2 cells. In contrast, LEE pretreatment significantly increased the level of Nrf2 and its downstream genes. LEE protects against TP-induced oxidative stress partly via the activation of Nrf2 pathway.

[Helicobacter pylori infection and associated risk factors in Chengdu].

OBJECTIVE: To investigate the prevalence of Helicobacter pylori (Hp) infection in Chengdu and its risk factors. METHODS: Permanent residents in Chengdu who took routine health examinations in the West China Hospital of Sichuan University were recruited in the study. All participants received 14C-urea breath test (14C-UBT) for the diagnosis of Hp infection. A random sample (n = 500) was drawn for a questionnaire survey. RESULTS: Of 8,365 participants, 53.1% were diagnosed with Hp infection. Higher prevalence was found in male participants (54.1%) than in female participants (51. 7%, P < 0.05). More than half of those older than 30 years old were infected. Of the 500 people who were invited to participate in the questionnaire survey, 497 returned questionnaires eligible for analyses. Compared with Han, Tibetans had a higher level of Hp infection (74.2% vs. 48.6%, P < 0.05). Higher levels of Hp infection were also found in those with vomiting symptoms (63.4% vs. 52.2%, P < 0.05). The participants who ate raw garlic had a lower level of Hp infection than those who did not eat raw garlic (52.6% vs. 67.9%, P < 0.05). CONCLUSION: The prevalence of Hp infection in Chengdu is high. Male, Tibetan and vomiting symptoms are predictors of Hp infection, while raw garlic seems to reduce the infection.

[Association of chronic obstructive pulmonary disease and lung cancer].

OBJECTIVE: To investigate the association between chronic obstructive pulmonary disease (COPD) and lung cancer. METHODS: A case-control study was undertaken, with 180 cases of lung cancer and 200 cases of controls. RESULTS: The odd of lung cancer was higher in patients with COPD, emphysema, chronic bronchitis, and pulmonary tuberculosis (P < 0.05). The odd of lung cancer increased significantly in patients with a family history of lung cancer or COPD (P < 0.05). The odd of lung cancer also increased when forced expiratory volume in one second (FEV1) < 80%. CONCLUSION: Patients with COPD or a family history of COPD have higher risk of lung cancer.

HPLC-DAD method for comprehensive quality control of Semen Strychni.

CONTEXT: Semen Strychni is the seed of Strychnos nux-vomica L. (Loganiaceae). Its quality control procedure remains an issue since previous reports only focused on Strychnos alkaloids. To the best of our knowledge, chlorogenic acid (a phenolic acid) and loganin (an iridoid glycoside) are selected for the first time as marker constituents of quality control for Semen Strychni because of their bioactive activity correlating with therapeutic effects. OBJECTIVE: This study aimed to develop a simple and comprehensive quantity control method for Semen Strychni. MATERIALS AND METHODS: The optimal ultrasonic extraction procedure was carried out for 45 min using 50% aqueous methanol with 1% formic acid. The satisfactory chromatographic separation was achieved on an Ultimate LP-C18 column with gradient elution using acetonitrile and water containing 30 mmol/L ammonium acetate and 1% formic acid. The high performance liquid chromatography method with diode array detector was validated for linearity, limit of detection and quantification (LOQ), precision, repeatability, accuracy and stability. RESULTS: All the calibration curves showed good linearity (r(2) ≥ 0.999). The LOQ values for chlorogenic acid, loganin, strychnine, brucine, strychnine N-oxide and brucine N-oxide were 0.54, 0.83, 0.48, 0.50, 0.52 and 0.54 µg/mL, respectively. The method was reproducible with good accuracy in the range 95.6-104.4% and relative standard deviation (RSD) values less than 4.55%. The method was then applied to determine the components of the seed coat, seed leaf, endosperm and whole seed of Semen Strychni. CONCLUSION: This newly established method is validated as a simple and practical tool for authentication and quality control of Semen Strychni.

Triangular Ising antiferromagnets with quenched nonmagnetic impurities.

In a random spin system, the cooperation of randomness and frustration will lead to a spin-glass phase. However, in geometrically frustrated spin systems, quenched nonmagnetic impurities lift frustration locally. This makes randomness and frustration in these systems as competitors rather than cooperators. By mapping the dilute triangular Ising antiferromagnetic system to elastic array of noncrossing strings, we find that the nonmagnetic impurities in the spin system play roles of pinning centers in the string system. Calculation shows that in the ground state of this system, the spin-glass correlation is power-law decayed, quite different from the standard behavior of spin glass in which spin-glass correlation between two spins at infinite distance tends to a finite value. This indicates that triangular Ising antiferromagnets with quenched nonmagnetic impurities cannot be a spin glass. Instead, in the ground states, they present properties of vortex glass.

[Adaptive evolution of chemokines and their receptors genes].

Branch-Site Model is a statistical method for detecting molecular adaptation at individual along specific lineages. Not only the model could indicate whether genes in phylogeny under the positive selection or not, but also could forecast positive selection sites in these genes. The sites promote divergence and polymorphism of genes. Chemokines are chemotactic cytokines that can induce immune cells migration, conducting their functions via chemokine receptors. In the test, the molecular adaptation of chemokines and chemokines receptors genes had been analyzed by Branch-Site model. The results showed that only a few of genes, such as RANTES and CCR5, are in the course of positive selection. Several CCR5 positive selection sites are located in a region that involved in binding to receptor and its chemokine.

A small molecule compound inhibits AKT pathway in ovarian cancer cell lines.

BACKGROUND AND OBJECTIVE: Overactivation of AKT1 and gene amplification of AKT2 are frequently detected in ovarian cancer. Activated AKT kinases provide a cell survival signal that may confer resistance to apoptosis induced by conventional therapies in cancer cells. Therefore, development of potent inhibitors that block AKT pathway is an attractive therapeutic strategy for treating ovarian carcinoma. METHODS: Ovarian cancer cell lines, A2780, MDAH2774, OVCAR-8, Caov-3, and normal murine fibroblasts (NIH3T3) were used. Cells were treated with different doses of a non-peptide small molecule compound, 9-methoxy-2-methylellipticinium acetate (termed API-59-OME) that potentially inhibit AKT pathway. Kinase assays and the phosphorylation of AKT, GSK-3alpha/beta, PDK1, ERK1/2, SGK, p38, FAK, EGFR, JAK2, PKC isoforms, and the cleavage of poly (ADP-ribose) polymerase (PARP) were examined in treated and untreated cell lines. Further, cells treated with API-59-OME were analyzed for induction of apoptosis using sub-G1 profile with propidium iodide staining. RESULTS: API-59-OME inhibited AKT kinase activity but did not inhibit ERK or JNK kinase activities in A2780, MDAH2774, and OVCAR-8 cell lines. API-59-OME did not reduce phosphorylation of other protein kinases in these cell lines. API-59-OME induced apoptosis and the cleavage of PARP in A2780, MDAH2774, and OVCAR-8 ovarian cancer cell lines that express elevated levels of phosphorylated AKT. In contrast, in Caov-3 and NIH3T3 cell lines, which lack constitutive AKT activity, API-59-OME only had minimal effect to induce apoptosis. CONCLUSION: These data suggest that API-59-OME may be a potent agent to target constitutively activated AKT pathway in ovarian cancer cells.

Modulation of Janus kinase 2 by p53 in ovarian cancer cells.

The constitutive activation of the Janus kinase 2 (JAK2) and mutation of the p53 tumor suppressor are both detected in human cancer. We examined the potential regulation of JAK2 phosphorylation by wild-type (wt) p53 in human ovarian cancer cell lines, Caov-3 and MDAH2774, which harbor mutant form of p53 tumor suppressor gene and high levels of phosphorylated JAK2. The wt p53 gene was re-introduced into the cells using an adenovirus vector. In addition to wt p53, mutant p53 22/23, mutant p53-175, and NCV (negative control virus) were introduced into the cells in the control groups. Expression of wt p53, but not that of p53-175 mutant, diminished JAK2 tyrosine phosphorylation in MDAH2774 and Caov-3 cell lines. Expression of wt p53 or p53 22/23 mutant did not cause a reduction in the phosphorylation of unrelated protein kinases, ERK1 and ERK2 (ERK1/2). The inhibition of JAK2 tyrosine phosphorylation can be reversed by tyrosine phosphatase inhibitor, sodium orthovanadate. Protein tyrosine phosphatase 1-B levels increased with introduction of wt p53 and may be involved in the dephosphorylation of JAK2. These findings present a possible p53-dependent cellular process of modulating JAK2 tyrosine phosphorylation in ovarian cancer cell lines.

[Transcriptional inactivation of RASSF1A in lung cancer and its clinical significance].

OBJECTIVE: To explore the relationship between the transcriptional expression of RASSF1A (Ras association domain family 1A gene) and oncogenesis and development of lung cancer. METHOD: RT-PCR was used to detect the expression of RASSF1A mRNA in 47 human lung cancer tissues and matched 47 non-cancer tissues. RESULTS: (1). The RASSF1A mRNA was identified in all non-cancer tissues but not found in 53.2% carcinoma tissues. (2). The rate of loss of RASSF1A mRNA was significantly higher in the patients with positive lymph node metastasis (70.4%) than in those without lymph node metastasis (30.0%) (P < 0.01). (3). Much more frequent in advanced tumor tissues, loss of RASSF1A mRNA was correlated with TNM stage (P < 0.05). (4). No significant association of abnormal RASSF1A expression was identified with histological type, differentiation grade of tumors or age, sex, and smoking index of the patients (all P > 0.05). CONCLUSION: RASSF1A is one of the new candidates of tumor suppressor genes. Loss or abnormal down-regulation of RASSF1A mRNA is a frequent event in lung carcinogenesis, which may play an important role in the malignant progression and prognosis of lung cancer.