Mining of Gene Modules and Identification of Key Genes for Early Diagnosis of Gastric Cancer

Gastric cancer (GC) is one of the most common malignant tumors with high incidence and mortality rates. Most patients with GC are not diagnosed until the advanced stage of cancer or during tumor screening, resulting in missing the best treatment time. This study identified key modules and hub genes associated with GC by weighted gene co-expression network analysis (WGCNA). The "limma" package in R was used to identify differentially expressed genes (DEGs) in GC samples from TCGA, and a total of 4892 DEGs were identified. GO enrichment and KEGG pathway enrichment analyses were conducted to detect the related pathways and functions of DEGs. These DEGs were primarily associated with extracellular matrix organization, DNA replication, cell cycle, and p53 signaling pathway. Gene modules associated with clinical characteristics were identified with WGCNA in tumor and normal samples. Six gene modules were obtained in the WGCNA network, of which two modules were significantly correlated with GC. Hub genes of key modules were identified using survival analysis and expression analysis. Finally, one-way ANOVA was used to explore the relationship between hub gene expression in normal tissues and different pathological stages of GC. Through survival and expression analysis, a total of 19 genes with good prognosis and significantly differential expressed were identified. The hub genes were significantly differential expressed in normal tissues and different pathological stages of GC, indicating that these genes have important diagnostic value for early GC and can be used as auxiliary indicators in the diagnosis of early GC.

Protective effects of sodium ferulate on flap transplantation via anti-inflammatory modulation and oxidative stress inhibition

Ischemia-reperfusion injury (IRI) has brought attention to flap failure in reconstructive surgery. To improve the prognosis of skin transplantation, we performed experimental IRI by surgical obstruction of blood flow and used sodium ferulate (SF) to prevent IRI in rats. After SF treatment, the morphological and histological changes of the skin flaps were observed by H&E and Masson's trichrome staining. We also detected the expression levels of COX-1, HO-1, and Ki67 by immunohistochemical and western blot analysis. Moreover, enzyme-linked immunosorbent assay was used to identify the content of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), malondialdehyde (MDA), and nitric oxide (NO) in peripheral blood and skin tissue. Compared with the model group, SF treatment significantly improved the recovered flap area (%) and promoted collagen synthesis. Cyclooxygenase-2 (COX-2) expression was significantly inhibited by heme oxygenase-1 (HO-1) induction after SF treatment. Furthermore, SF significantly inhibited the levels of TNF-α in peripheral blood, MPO and MDA in the skin tissue, and the increased synthesis of NO. Our results showed the protective effects of SF on IRI after flap transplantation and we believe that the protective effects of SF was closely related to the alleviation of the inflammatory response and the inhibition of the oxidative stress injury.

Cantharidin suppressed breast cancer MDA-MB-231 cell growth and migration by inhibiting MAPK signaling pathway

As an active constituent of the beetle Mylabris used in traditional Chinese medicine, cantharidin is a potent and selective inhibitor of protein phosphatase 2A (PP2A) that plays a crucial role in cell cycle progression, apoptosis, and cell fate. The role and possible mechanisms exerted by cantharidin in cell growth and metastasis of breast cancer were investigated in this study. Cantharidin was found to inhibit cell viability and clonogenic potential in a time- and dose-dependent manner. Cell cycle analysis revealed that cell percentage in G2/M phase decreased, whereas cells in S and G1 phases progressively accumulated with the increasing doses of cantharidin treatment. In a xenograft model of breast cancer, cantharidin inhibited tumor growth in a dose-dependent manner. Moreover, high doses of cantharidin treatment inhibited cell migration in wound and healing assay and downregulated protein levels of major matrix metalloproteinases (MMP)-2 and MMP-9. MDA-MB-231 cell migration and invasion were dose-dependently inhibited by cantharidin treatment. Interestingly, the members of the mitogen-activated protein kinase (MAPK) signaling family were less phosphorylated as the cantharidin dose increased. Cantharidin was hypothesized to exert its anticancer effect through the MAPK signaling pathway. The data of this study also highlighted the possibility of using PP2A as a therapeutic target for breast cancer treatment.

Angiotensin-(1-7) relieved renal injury induced by chronic intermittent hypoxia in rats by reducing inflammation, oxidative stress and fibrosis

We aimed to study the renal injury and hypertension induced by chronic intermittent hypoxia (CIH) and the protective effects mediated by angiotensin 1-7 [Ang(1-7)]. We randomly assigned 32 male Sprague-Dawley rats (body weight 180-200 g) to normoxia control, CIH, Ang(1-7)-treated normoxia, and Ang(1-7)-treated CIH groups. Systolic blood pressure (SBP) was monitored at the start and end of each week. Renal sympathetic nerve activity (RSNA) was recorded. CTGF and TGF-β were detected by immunohistochemistry and western blotting. Tissue parameters of oxidative stress were also determined. In addition, renal levels of interleukin-6, tumor necrosis factor-α, nitrotyrosine, and hypoxia-inducible factor-1α were determined by immunohistochemistry, immunoblotting, and ELISA. TUNEL assay results and cleaved caspase 3 and 12 were also determined. Ang(1-7) induced a reduction in SBP together with a restoration of RSNA in the rat model of CIH. Ang(1-7) treatment also suppressed the production of reactive oxygen species, reduced renal tissue inflammation, ameliorated mesangial expansion, and decreased renal fibrosis. Thus, Ang(1-7) treatment exerted renoprotective effects on CIH-induced renal injury and was associated with a reduction of oxidative stress, inflammation and fibrosis. Ang(1-7) might therefore represent a promising therapy for obstructive sleep apnea-related hypertension and renal injury.

Angiotensin- (1-7) inhibits inflammation and oxidative stress to relieve lung injury induced by chronic intermittent hypoxia in rats

Obstructive sleep apnea is associated with inflammation and oxidative stress in lung tissues and can lead to metabolic abnormalities. We investigated the effects of angiotensin1–7 [Ang-(1–7)] on lung injury in rats induced by chronic intermittent hypoxia (CIH). We randomly assigned 32 male Sprague-Dawley rats (180–200 g) to normoxia control (NC), CIH-untreated (uCIH), Ang-(1–7)-treated normoxia control (N-A), and Ang-(1–7)-treated CIH (CIH-A) groups. Oxidative stress biomarkers were measured in lung tissues, and expression of NADPH oxidase 4 (Nox4) and Nox subunits (p22phox, and p47phox) was determined by Western blot and reverse transcription-polymerase chain reaction. Pulmonary pathological changes were more evident in the uCIH group than in the other groups. Enzyme-linked immunosorbent assays and immunohistochemical staining showed that inflammatory factor concentrations in serum and lung tissues in the uCIH group were significantly higher than those in the NC and N-A groups. Expression of inflammatory factors was significantly higher in the CIH-A group than in the NC and N-A groups, but was lower than in the uCIH group (P<0.01). Oxidative stress was markedly higher in the uCIH group than in the NC and N-A groups. Expression of Nox4 and its subunits was also increased in the uCIH group. These changes were attenuated upon Ang-(1–7) treatment. In summary, treatment with Ang-(1-7) reversed signs of CIH-induced lung injury via inhibition of inflammation and oxidative stress.

Effects of fluoride on proliferation and mineralization in periodontal ligament cells in vitro

Fluoride, which is often added to toothpaste or mouthwash in order to protect teeth from decay, may be a novel therapeutic approach for acceleration of periodontal regeneration. Therefore, we investigated the effects of fluoride on proliferation and mineralization in human periodontal ligament cells in vitro. The periodontal ligament cells were stimulated with various concentrations of NaF added into osteogenic inductive medium. Immunohistochemistry of cell identification, cell proliferation, alkaline phosphatase (ALP) activity assay, Alizarin red S staining and quantitative real-time-polymerase chain reaction (RT-PCR) were performed. Moderate concentrations of NaF (50-500 μmol/L) had pro-proliferation effects, while 500 μmol/L had the best effects. ALP activity and calcium content were significantly enhanced by 10 μmol/L NaF with osteogenic inductive medium. Quantitative RT-PCR data varied in genes as a result of different NaF concentrations and treatment periods. We conclude that moderate concentrations of NaF can stimulate proliferation and mineralization in periodontal ligament cells. These in vitro findings may provide a novel therapeutic approach for acceleration of periodontal regeneration by addition of suitable concentrations of NaF into the medication for periodontitis treatment, i.e., into periodontal packs and tissue patches.

Impact of System Factors and Modifiable ICU Interventions on the Outcome of Cardio-pulmonary Resuscitation in PICU.

Objectives: To assess the impact of system factors and modifiable interventions on outcome of cardiac arrest in a pediatric intensive care unit. Design: Retrospective medical record review. Setting: Pediatric intensive care unit of a hospital in China. Participants: Children (age<14 yrs) who had cardiac arrest within our PICU over a period of two years. Results: Sixty-one of the 94 cardiac arrest events were successfully resuscitated. There was no significant association between personal and unit factors with immediate outcomes in our unit. The rate of unsuccessful resuscitation in sedated patients and those without sedation was 26% and 50%, respectively. Unsuccessful resuscitation occurred in 19% of patients who were on positive pressure ventilation as compared with 74% for those without positive pressure ventilation. Arrests which had resuscitation attempts that lasted more than 30 min had 135-fold higher odds of unsuccessful outcome. 78% of patients who received base supplement at the time of arrest had unsuccessful resuscitation compared with 21% for those without base supplement. Conclusions: Our data shows no impact of system factors on the outcome of cardio-pulmonary resuscitation in our PICU. Prearrest sedation in pediatric critical ill patients might be beneficial to the outcome of cardiac arrests.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

Outcome analysis of benign vocal cord tumors treated by laryngeal endoscopy under low temperature-controlled radiofrequency.

OBJECTIVE: This study aimed to evaluate the outcome of benign vocal cord tumors treated using a laryngeal endoscopy under low temperature‑controlled radiofrequency and to elucidate the application of a dynamic laryngoendoscopy in the operation. MATERIALS AND METHODS: 85 patients with benign vocal cord tumors were treated by laryngeal endoscopy under low temperature‑controlled radiofrequency from September 2011 to October 2013. A XION electronic dynamic laryngoendoscopy (Germany) was used to observe curative effects 3 months after operation. Wave images were recorded with larynx‑wave recording software to analyze tumor characteristics. RESULTS: Among the 85 patients, 81 showed smooth surface of operation wounds without any residue. The mucosal wave was also basically normal. Sound was generally recovered after 1–3 months. Three cases presented improved pronunciation function after the operation, whereas 1 patient with residual tumor at the front of vocal chords underwent another operation after 6 months.CONCLUSION: Low temperature‑controlled radiofrequency exhibited many advantages, including minimal trauma, minimal bleeding, high safety, and few complications. Moreover, treatment of benign vocal cord tumors with a laryngeal endoscopy presented satisfactory outcomes. Therefore, this technology has broad application prospects.

Detection of acute cerebral hemorrhage in rabbits by magnetic induction

Acute cerebral hemorrhage (ACH) is an important clinical problem that is often monitored and studied with expensive devices such as computed tomography, magnetic resonance imaging, and positron emission tomography. These devices are not readily available in economically underdeveloped regions of the world, emergency departments, and emergency zones. We have developed a less expensive tool for non-contact monitoring of ACH. The system measures the magnetic induction phase shift (MIPS) between the electromagnetic signals on two coils. ACH was induced in 6 experimental rabbits and edema was induced in 4 control rabbits by stereotactic methods, and their intracranial pressure and heart rate were monitored for 1 h. Signals were continuously monitored for up to 1 h at an exciting frequency of 10.7 MHz. Autologous blood was administered to the experimental group, and saline to the control group (1 to 3 mL) by injection of 1-mL every 5 min. The results showed a significant increase in MIPS as a function of the injection volume, but the heart rate was stable. In the experimental (ACH) group, there was a statistically significant positive correlation of the intracranial pressure and MIPS. The change of MIPS was greater in the ACH group than in the control group. This high-sensitivity system could detect a 1-mL change in blood volume. The MIPS was significantly related to the intracranial pressure. This observation suggests that the method could be valuable for detecting early warning signs in emergency medicine and critical care units.

Induced maturation of hepatic progenitor cells in vitro

Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

General practice education and training in southern China: recent development and ongoing challenges under the health care reform

China has launched a general practice (GP)-orientated primary care reform in 2009 to develop a more productive, coordinated, and cost-effective system to maintain and improve the health and wellbeing of one-fifth of the world population. The restructure of the health care system with a focus on primary care requires practitioners working on GP as gatekeepers for service delivery that is responsive to the needs of people. It is particularly prioritised to establish a sound education and training system to ensure that the competencies of practitioners are aligned with local health care needs. This article aims to provide a brief review of the development of GP, including exemplary model of education and training currently implemented in southern China, as well as the challenges to be addressed in the next step. There is a shortage of well-trained and qualified general practitioners in China where more than half of the licensed clinicians in primary care are educated below the undergraduate level. Although there is a stepwise increase in recognition that the capacity of GP is pivotal to the success of primary care development in China, challenges coming from resource restriction, rural and urban disparity, social attitude, and community involvement are highlighted as major bottlenecks that currently hinder the rapid development of GP in China. Supportive policy and guidelines are necessary to build up strong GP recognition and ensure adequate resources to underpin a robust primary care system to deliver affordable and effective health care services for the world’s largest population. It might share some similar experiences with other countries that are struggling to develop a GP-based primary care system.

The survey of HIV/AIDS related knowledge, attitude and behaviour among migrant peasant workers in Middle City in China / Encuesta de conocimientos, actitudes, y comportamientos en relación con el VIH/SIDA entre los trabajadores campesinos migratorios de la Ciudad Media en China

OBJECTIVES: To explore HIV/AIDS related knowledge, attitude and behaviour among migrant peasant workers in Changsha, which is a inland city in central China, and to provide evidence that educating this population may help to prevent the spread of HIV/AIDS in China. METHODS: A total of 1782 migrant peasant workers participated in the study. A questionnaire was used to determine their knowledge ofHIV/AIDS and related attitude and behaviour items. RESULTS: The majority of the respondents had a rather low level of overall HIV/AIDS knowledge and its prevention, had what constituted risky behaviour in this regard and a generally low acceptance of HIV-infected persons. Furthermore, their behaviour and attitude were analysed with respect to their educational level, occupational class and score ofHIV/AIDS related knowledge. CONCLUSIONS: The results of this investigation will help healthcare professionals develop a sensitive and specific educational programme for migrant peasant workers. Educating this population will be a very important aspect ofHIV/AIDS prevention in China.

OBJETIVOS: Explorar los conocimientos, las actitudes, y el comportamiento en relación con el VIH/SIDA, entre los trabajadores campesinos migratorios en Changsha - una ciudad del interior en China central -y proporcionar evidencias en cuanto al hecho de que educar a esta población puede ayudar a prevenir la diseminación del VIH/SIDA en China. MÉTODOS: Un total de 1782 trabajadores campesinos migratorios participó en el estudio. Se usó un cuestionario a fin de determinar sus conocimientos sobre el VIH/SIDA, así como aspectos relacionados con la actitudy el comportamiento hacia este último. RESULTADOS: La mayoría de los encuestados tenían un nivel bastante bajo de conocimientos generales sobre el VIH/SIDA y su prevención, poseían lo que pueden considerarse como una conducta riesgosa al respecto, y manifestaban una aceptación generalmente baja de las personas infectadas con el VIH. Además, se analizaron su comportamiento y actitud con respecto a su nivel educacional, clase ocupacional y puntuación en cuanto a conocimientos en relación con el VIH/SIDA. CONCLUSIONES: Los resultados de esta investigación ayudarán a los profesionales de la salud a desarrollar un programa educativo sensible y específico para los trabajadores campesinos migratorios. Educar a esta población será un aspecto muy importante para la prevención del VIH/SIDA en China.

Nationwide survey of human parasite in China.

A nationwide survey of human parasites in China was conducted during 1988-1992, with a coverage of 30 provinces/autonomous regions/municipalities (P/A/M). A total of 2,848 pilot sites in 726 counties were selected by random sampling, and 1,477,742 individuals residing on were surveyed by fecal examination. The status of paragonimiasis, hydatid diseases, cysticercosis and trichinellosis were summarized through data review. The overall infection rate of intestinal parasites was 62.6% whereas at provincial level, the highest infection rate (94.7%) was recovered in Hainan, and the lowest (17.5%) in Heilong-jiang. A high proportion (43.3%) of polyparasitism among the infected population (882,080) was revealed. Altogether 56 species of parasites comprising protozoa (19), trematode (16), cestodes (8), nematodes (12) and thorny-headed worm (1) were discovered. During the survey a new species and several new records were documented. The number of the population infected with common intestinal parasites was estimated. The diversities of parasite distribution were noted in different nationalities as well as in varied occupations.

Antibody detection ELISAS for malaria diagnosis.

Parasite extracts of Plasmodium falciparum and P. chabaudi and three synthetic peptides from the P. falciparum MSA2 merozoite antigen were tested for suitability as antigens in an antibody detection ELISA using sera from malaria patients in Brisbane. The P. chabaudi extract was superior to P. falciparum extract for detecting P. vivax cases, while for P. falciparum cases the two parasite extracts were equivalent. Single peptide antigens were generally less sensitive than parasite extracts; however, peptides G3 and G7 were more sensitive than parasite extracts in detecting first attacks of P. vivax. Examination of isotype specific responses demonstrated that this may be explained by higher IgG responses to these peptides in first than in subsequent P. vivax attacks. Because of the differing antibody specificities in primary and secondary P. falciparum and P. vivax cases, the best sensitivity was achieved by using the combined results of assays with three antigens: P. chabaudi, peptide G3 and peptide G7. The combined sensitivity was 77.1% for P. falciparum and 88.6% for P. vivax acute cases with 91.1% specificity.