Beneficial Effects of High-Intensity Interval Training and Dietary Changes Intervention on Hepatic Fat Accumulation in HFD-Induced Obese Rats.

Lifestyle intervention encompassing nutrition and physical activity are effective strategies to prevent progressive lipid deposition in the liver. This study aimed to explore the effect of dietary change, and/or high-intensity interval training (HIIT) on hepatic lipid accumulation in high fat diet (HFD)-induced obese rats. We divided lean rats into lean control (LC) or HIIT groups (LH), and obese rats into obese normal chow diet (ND) control (ONC) or HIIT groups (ONH) and obese HFD control (OHC) or HIIT groups (OHH). We found that dietary or HIIT intervention significantly decreased body weight and the risk of dyslipidemia, prevented hepatic lipid accumulation. HIIT significantly improved mitochondrial fatty acid oxidation through upregulating mitochondrial enzyme activities, mitochondrial function and AMPK/PPARalpha/CPT1alpha pathway, as well as inhibiting hepatic de novo lipogenesis in obese HFD rats. These findings indicate that dietary alone or HIIT intervention powerfully improve intrahepatic storage of fat in diet induced obese rats. Keywords: Obesity, Exercise, Diet, Mitochondrial function, Lipid deposition.

Clinical correlation between volume of placental lakes and non-reassuring fetal status: a retrospective cohort study.

OBJECTIVE: This study evaluated the correlation between placental lakes and non-reassuring fetal status. SUBJECTS AND METHODS: We analyzed data from pregnant women who underwent fetal echocardiography at the Fujian Maternity and Child Health Hospital. Women with singleton pregnancies at a gestational age of 20-24 weeks were included. Sociodemographic and clinical data were collected. Pregnant women with (case group) and without (control group) placental lakes were screened, and their placental Doppler ultrasound data and pregnancy outcome were recorded. Univariate and multivariable analyses were done to evaluate the correlation between the volume of placental lakes and the non-reassuring fetal status. RESULTS: A total of 1,728 pregnant women (156 with placental lakes) were included in this study. There were no significant differences in age of delivery and BMI between the pregnant women with placental lakes and the control group. The non-reassuring fetal status rate in the case group was higher than that in the control population, without statistical significance (5.8% vs. 3.5%, p=0.226). Subgroup analysis showed that a higher volume of placental lakes was positively associated with non-reassuring fetal status risk, with an odds ratio (OR) (95% CI) of 1.90 (1.29-2.66) (p for trend < 0.001). This positive correlation persisted even after adjustment for confounding factors. CONCLUSIONS: Taken together, our analyses demonstrated a graded increase in the non-reassuring fetal status rate with increased volume of placental lakes. Thus, robust clinical monitoring of placental lakes would help in timely detection of non-reassuring fetal status.

Incidence and familial clustering of infantile haemangiomas: a multicentre study.

BACKGROUND: The exact incidence of infantile haemangiomas (IH) in the Chinese population is still unknown. A positive family history of IH was considered as a risk factor for the development of IH. OBJECTIVES: This study aims to investigate the incidence of IH in the Chinese population and the mechanism of family history increases the risk for IH development. METHODS: A total of 2489 women and their newborns were enrolled in the prospective study. All newborns were followed up for 12 months to determine whether they developed IH. In addition, 213 IH probands and their 174 siblings were enrolled in the study. The incidence of IH in siblings of the IH probands was investigated. Information regarding risk factors for IH and demographic data were collected on all children. RESULTS: Of the 2572 newborns, 58 IH were identified in 56 (2.2%) newborns. The majority of IH were located on the trunk (46.6%). Siblings of the IH probands were at increased risk for the development of IH (P = 0.024, relative risk 2.451), and the occurrence of prenatal risk factors for IH(P = 0.003) compared with the general population. CONCLUSIONS: Our study showed that the incidence of IH is 2.2% in the Chinese population. Siblings of the individuals with IH were at increased risk for the development of IH may be related to the family clustering of prenatal risk factors for IH. Further exploration of the mechanisms and common features of these prenatal risk factors may help to disclose the origin and pathogenesis of IH.

MiR-16 inhibits proliferation of cervical cancer cells by regulating KRAS.

OBJECTIVE: The aim of this study was to explore the effects of micro ribonucleic acid (miR)-16 on the proliferation and apoptosis of cervical cancer (CC) cells and its related regulatory mechanism. MATERIALS AND METHODS: The downstream regulatory targets of miR-16 were analyzed based on the miRNA online database. HCC94 cells were selected as experimental objects. Subsequently, the cells were transfected with miR-16 mimic (miR-16 mimic group), miR-16 small interfering RNA (siRNA) (miR-16 siRNA group) and only Lipofectamine 2000 transfection reagent [blank control group and miR-16 normal control (NC) group]. The expression level of miR-16 in HCC94 cells was measured via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Cell counting kit-8 (CCK-8) assay, 5-Ethynyl-2'-deoxyuridine (EdU) staining assay and flow cytometry were then conducted to detect the effects of miR-16 on the viability, proliferation and apoptosis of HCC94 cells, respectively. Additionally, the effect of miR-16 on the protein expression level of Kirsten rat sarcoma viral oncogene homolog (KRAS) in HCC94 cells was determined via Western blotting. RESULTS: MiRNA online database analysis showed that KRAS was the downstream target of miR-16. Compared with miR-16 NC group, the viability and proliferation ability of HCC94 cells increased significantly in miR-16 siRNA group but decreased significantly in miR-16 mimic group (p<0.05). However, the apoptosis rate evidently declined in miR-16 siRNA group while increased remarkably in miR-16 mimic group (p<0.05). In addition, the protein expression level of KRAS in HCC94 cells was significantly higher in miR-16 siRNA group but significantly lower in miR-16 mimic group when compared with miR-16 NC group (p<0.05). CONCLUSIONS: MiR-16 is lowly expressed in HCC94 cells. Moreover, highly expressed miR-16 represses the viability and proliferation of HCC94 cells and promotes their apoptosis by targeted regulation on KRAS.

MiR-376b-3p functions as a tumor suppressor by targeting KLF15 in non-small cell lung cancer.

OBJECTIVE: The aim of this study was to explore the regulatory effects of micro ribonucleic acid (miR)-376b-3p on proliferation and apoptosis of non-small cell lung cancer (NSCLC) cells by targeting Kruppel-like factor 15 (KLF15) and its mechanism of action. PATIENTS AND METHODS: The expression of miR-376b-3p in NSCLC and para-carcinoma normal tissues, as well as NSCLC cell lines, was detected via quantitative Polymerase Chain Reaction (qPCR). The effects of miR-548-3p on the proliferation, cycle distribution, and apoptosis of NSCLC cells were detected via colony formation assay, flow cytometry, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, respectively. The interaction between miR-376b-3p and KLF15 was determined using Dual-Luciferase reporter gene assay. In vivo tumorigenic ability of NSCLC cells was studied using nude mouse tumorigenicity assay. Furthermore, the expression of Ki67 in tumor in nude mice was detected via immunohistochemistry. RESULTS: The expression of miR-376b-3p was significantly downregulated in NSCLC tissues when compared with para-carcinoma normal tissues (p<0.05). MiR-376b-3p was lowly expressed in NSCLC cells as well (p<0.05). After overexpression of miR-376b-3p, the proliferation ability of NSCLC cells remarkably declined (p<0.05). The apoptosis rate rose, and cell cycle was arrested in the G1/G0 phase. Dual-Luciferase reporter gene assay confirmed that miR-376b-3p could specifically bind to KLF15 3'UTR to regulate the expression activity of KLF15. After overexpression of miR-376b-3p, tumor volume and weight were significantly reduced in tumor-bearing mice (p<0.05). CONCLUSIONS: MiR-376b-3p plays an important role in the occurrence and development of NSCLC, which affects the proliferation and apoptosis of NSCLC cells by targeting KLF15.

FKBP51 acts as a biomarker of early metastasis and is related to carmustine sensitivity in human glioma cells.

OBJECTIVE: Given that FK506 binding protein 51 (FKBP51) is upregulated in multiple cancers, we designed the present study to characterize its role as well as underlying regulatory mechanisms in glioma in the presence and absence of the chemotherapeutic carmustine (BCNU). MATERIALS AND METHODS: Through lentiviral overexpression and shRNA knockdown of FKBP51, we examined the effects on BT325 glioma cell proliferation, migration and invasion using quantitative reverse transcription PCR (qRT-PCR), CCK-8 assay, flow cytometry, and transwell assay. RESULTS: The upregulation of FKBP51 resulted in significantly decreased BT325 cell proliferation and cell viability, cell cycle arrest, reduced BCNU chemosensitivity and AKT pathway inactivation. However, FKBP51-overexpressed BT325 cells showed enhanced migration and invasion, which was supported by corresponding increase in phosphorylated IKKα (p-IKKα), MMP-2, and MMP-9 levels, as well as increased NF-κB p65 nuclear translocation. By contrast, FKBP51-suppressed BT325 cells showed excessive proliferation and BCNU resistance due to increased p-AKT activation and attenuated migration and invasion. CONCLUSIONS: We demonstrated that the effects of FKBP51 on BT325 glioma cell proliferation, migration, invasion and BCNU chemosensitization are modulated via the AKT and NF-κB pathways. Furthermore, our findings suggest the potential of FKBP51 as a prognostic glioma biomarker and an indicator of patient response to chemotherapy.

Effect of strontium ranelate on rabbits with steroid-induced osteonecrosis of femoral head through TGF-ß1/BMP2 pathway.

OBJECTIVE: To study the effect of strontium ranelate (SR) on steroid-induced osteonecrosis of the femoral head (SIONFH) in rabbits and its regulatory mechanism. MATERIALS AND METHODS: The ONFH model was established in 30 rabbits using steroid and they were randomly divided into Control group, Model group, and SR group. After SR intervention, the rabbits were sacrificed and sampled. The pathological injury of the femoral head in each group was detected via hematoxylin-eosin (HE) staining, the level of vascular endothelial growth factor (VEGF) in the femoral head in each group was detected via enzyme-linked immunosorbent assay (ELISA). The messenger ribonucleic acid (mRNA) and protein expression levels of transforming growth factor-ß1 (TGF-ß1), as well as the bone morphogenetic protein 2 (BMP2) in the femoral head in each group, were determined using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting. RESULTS: The rabbit model of SIONFH was successfully established. Compared with Control group, the Model group had a severer pathological injury of the femoral head, a lower level of VEGF in the femoral head, significantly decreased mRNA and protein levels of TGF-ß1 and BMP2. Compared with Model group, the SR group had markedly improved pathological injury of the femoral head, a higher level of VEGF in the femoral head, significantly increased mRNA and protein levels of TGF-ß1, as well as BMP2. CONCLUSIONS: SR can remarkably improve the pathological injury of the femoral head and increase the expression of VEGF in SIONFH rabbits, whose potential mechanism may be related to the activation of the TGF-ß1/BMP2 signaling pathway.

MicroRNA-544 inhibits inflammatory response and cell apoptosis after cerebral ischemia reperfusion by targeting IRAK4.

OBJECTIVE: Stroke remains the most common malignant cerebrovascular event in the world. The correlation between the expression of miR-544 and the degree of cerebral ischemia reperfusion (CIR) injury has not been well recognized in recent years. This study focuses on the effect of miR-544 on inflammation and apoptosis after CIR. PATIENTS AND METHODS: Plasma expression of miR-544 in ischemic stroke (IS) patients and healthy controls was determined by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). The effects of miR-544 on cerebral infarction and neurological deficits were verified in vitro by tail vein injection of Ago-miR-544. Western blotting was utilized to examine protein expressions of key proteins involving in inflammation and apoptosis in mouse brain. Western blotting, immunofluorescence staining and luciferase assays were used to demonstrate whether miR-544 influences the expression of interleukin-1 receptor-associated kinase 4 (IRAK4), downstream inflammatory and apoptosis-related proteins. RESULTS: MiR-544 was found decreased in peripheral blood of IS patients compared with healthy controls. MiR-544 has been shown to relieve neurological deficits and reduce the volume of cerebral infarction in mice. Overexpression of miR-544 ameliorated the inflammation and apoptotic responses in brain tissue after ischemia reperfusion by down-regulating the expression of IRAK4, whereas the low expression was opposite in vivo and in vitro. CONCLUSIONS: We found that miR-544 may participate in controlling inflammation and apoptosis after ischemia-reperfusion by targeting IRAK4, providing possible diagnostic indicators and therapeutic targets for IS.

Correlations of neuronal apoptosis with expressions of c-Fos and c-Jun in rats with post-ischemic reconditioning damage.

OBJECTIVE: Transcription factors (c-Fos and c-Jun) have been considered to play roles in the initiation of programmed nerve cell death. However, the roles of c-Fos and c-Jun protein expressions in neuronal apoptosis of rats with post-ischemic reconditioning damage were not clarified. Therefore, the aim of this study was to investigate the correlations of protein expressions of c-Fos and c-Jun with neuronal apoptosis of rats with post-ischemic reconditioning damage. MATERIALS AND METHODS: Rat models of post-ischemic reconditioning were established firstly. Then, apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay, and the gene expression levels of apoptosis-related proteins [cytochrome c (Cyt c), B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax)] were detected by reverse transcription-polymerase chain reaction (RT-PCR). Lastly, Western blotting was used to determine the protein expression levels of c-Fos and c-Jun, and the expressions of c-Fos and c-Jun in brain tissues of models were measured by immunohistochemistry. RESULTS: Treatment group had significantly increased malonaldehyde (MDA) level and significantly decreased superoxide dismutase (SOD) activity in rat cortex compared with those in control group (p<0.05). The number of TUNEL positive cells in the right cortex of rats in the treatment group was clearly higher than that in control group. Among them, post-ischemic reperfusion group had reduced level of Bax in the cytoplasm, but increased Bax level in the mitochondrion, and lowered expression level of Bcl-2 in both mitochondrion and cytoplasm in comparison with control group. Dynamic detection results of c-Jun were in synchronization with those of apoptosis proteins, and maximum expression occurred at 24 h after treatment. CONCLUSIONS: c-Jun may play a role in the initiation of apoptotic cell death in these neurons.

Research on the nutrition and cognition of high-risk stroke groups in community and the relevant factors.

OBJECTIVE: To investigate the prevalence rate of nutritional risk in high-risk stroke groups in community, analyze its influencing factors, and analyze and compare the relationship between nutritional risk or malnutrition assessed by different nutritional evaluation methods and cognitive function, so as to provide the basis and guidance for clinical nutritional assessment and support. PATIENTS AND METHODS: A cross-sectional survey was performed for 1196 cases in high-risk stroke groups in community from December 2015 to January 2017. At the same time, the nutritional status of patients was evaluated using the mini nutritional assessment (MNA) and MNA-short form (MNA-SF), and the cognitive status of patients was evaluated using the mini-mental state examination (MMSE). Moreover, the relevant influencing factors of nutritional risk and MMSE score were analyzed and compared. RESULTS: High-risk stroke groups in community suffered from a high risk of malnutrition. MNA-SF had a higher specificity and lower false positive rate than MNA. Nutritional risk occurred more easily in high-risk stroke groups in community with a history of diabetes mellitus, less physical exercise or light manual labor, daily use of multiple drugs, and higher age. Those with a higher nutritional risk were more prone to cognitive impairment. High-risk stroke groups in community, complicated with hyperhomocysteinemia, daily use of three or more kinds of prescription drugs, and a previous history of stroke, were accompanied by cognitive impairment easily. CONCLUSIONS: MNA-SF can be used for the nutritional screening of high-risk stroke groups in community. For the high-risk stroke groups in community, the rational nutritional diet should be publicized, blood sugar should be controlled in a scientific manner and physical exercise should be moderately increased.

Immune complexes induce TNF-α and BAFF production from U937 cells by HMGB1 and RAGE.

OBJECTIVE: This study investigated the effects of immune complexes (ICs) on tumor necrosis factor α (TNF-α) and B cell-activating factor (BAFF) production from U937 cells and further explored the mechanism. MATERIALS AND METHODS: U937 cells were incubated with necrosis supernatant or systemic lupus erythematosus (SLE) sera alone, or their combination. The expression of TNF-α and BAFF was determined by Real-time polymerase chain reaction and enzyme-linked immunosorbent assay. High mobility group box protein 1(HMGB1) A-box was produced by gene recombination. HMGB1 A-box and anti-receptor for advanced glycation end products (RAGE) antibody were adopted in the blocking experiments. The importance of DNA for cytokine induction was investigated by DNase treatment. RESULTS: The combination of necrosis supernatant and SLE sera induced the expression of TNF-α and BAFF significantly increased compared to necrosis supernatant or SLE sera alone. Recombinant HMGB1 A-box protein was purified, and TNF-α and BAFF production, which were induced by this combination, was blocked via HMGB1 A-box and anti-RAGE antibody. Moreover, we found that DNA component is important for the immunostimulatory activity of this combination. CONCLUSIONS: ICs containing DNA can promote TNF-α and BAFF production in U937 cells, and this process can be mediated by HMGB1 and RAGE. One possible mechanism of increasing BAFF production in SLE is proposed in this study whereby B cell activation, antibody production and ICs stimulated monocytes may create a vicious cycle that leads to B cell hyperactivity, which can be of importance for SLE etiopathogenesis.

Propranolol inhibits the proliferation, migration and tube formation of hemangioma cells through HIF-1α dependent mechanisms

The aim of this study was to investigate the mechanism of propranolol on the regression of hemangiomas. Propranolol-treated hemangioma tissues were collected and the expression of hypoxia inducible factor-1α (HIF-1α) was examined. We also established HIF-1α overexpression and knockdown hemangioma cells, and determined the effects of HIF-1α on the hemangioma cells proliferation, apoptosis, migration and tube formation. Significantly increased HIF-1α level was found in the hemangioma tissues compared to that in normal vascular tissues, whereas propranolol treatment decreased the HIF-1α level in hemangioma tissues in a time- and dose-dependent manner. Moreover, propranolol treatment significantly decreased cell proliferation, migration and tube formation as well as promoted cell apoptosis in HIF-1α overexpression and knockdown hemangioma cells. Propranolol suppressed the cells proliferation, migration and tube formation of hemangioma cells through HIF-1α dependent mechanisms. HIF-1α could serve as a novel target in the treatment of hemangiomas.

Clinical significance of changes of expression of the Wnt/ß-catenin signaling pathway in renal clear cell carcinoma.

OBJECTIVE: To explore the changes and clinical significance of expression of ß-catenin in renal carcinoma. PATIENTS AND METHODS: We selected 46 patients with renal clear cell carcinoma who were hospitalized from May 2013 to March 2016 and healthy adults (controls) matched for age and body weight, who were hospitalized in the physical examination center of our hospital during the same period. Peripheral blood of patients and controls was drawn for ELISA. After surgery, renal carcinoma and normal peritumoral tissue samples were harvested for immunohistochemical staining, Western blot analysis and qRT-PCR was used to observe changes of ß-catenin expression in renal carcinoma tissues. RESULTS: Compared with controls, ELISA showed that there were significant differences in ß-catenin levels in peripheral blood of patients with renal carcinoma (p<0.05). Western blot and qRT-PCR showed that the expression levels of ß-catenin in renal carcinoma tissues were higher than in normal peritumoral tissues and the differences were statistically significant (p<0.05). Immunohistochemical staining showed that ß-catenin was increased significantly in renal carcinoma tissue compared with normal peritumoral tissues (p<0.05). CONCLUSIONS: ß-catenin expression was significantly increased in the tumor tissues of patients with renal carcinoma. The measurement of ß-catenin expression levels in peripheral blood from patients could be used for early diagnosis of renal carcinoma, which is of great clinical significance.

Diffusion-weighted imaging to differentiate metastatic from non-metastatic retropharyngeal lymph nodes in nasopharyngeal carcinoma.

OBJECTIVES: To evaluate the diagnostic value of diffusion-weighted MRI for differentiating metastatic from non-metastatic retropharyngeal lymph nodes (RLNs) in patients with nasopharyngeal carcinoma (NPC). METHODS: Untreated patients with NPC (n = 145) were scanned with both morphological MRI and diffusion-weighted imaging (DWI). RLNs (n = 335) were classified as metastatic on the basis of response to therapy as assessed on follow-up MRI. Morphological (short- and long-axial diameters) and functional [mean apparent diffusion coefficient (ADC) and minimum ADC values] parameters of the RLNs were derived from DWI and compared between metastatic and non-metastatic groups. A receiver operating characteristic curve and the area under the curve were used to evaluate the effectiveness of individual criteria and to generate threshold values to diagnose RLN metastases. RESULTS: Statistically significant differences between metastatic and non-metastatic RLNs were found for all four parameters derived from DWI (p < 0.001). At threshold values, accuracies of the ADC-based criteria (0.938 and 0.965 for mean and minimum ADC values, respectively) were greater than that of size-based criteria (0.838 and 0.809 for short- and long-axial diameters). The minimum ADC value at the threshold of 0.89 × 10(-3) mm(2) s(-1) was the most effective of all parameters in differentiating metastatic from non-metastatic RLNs with the sensitivity of 95.7%, specificity of 95.1% and accuracy of 96.5%. CONCLUSIONS: DWI is feasible for differentiating metastatic RLNs from non-metastatic nodes in patients with NPC with high accuracy, and the minimum ADC derived from DWI could serve as a standard clinical marker for disease status.

Relationship between leptin and chronic inflammatory state in uremic patients.

OBJECTIVE: To explore the relation between high leptin and inflammation in uremic patients. PATIENTS AND METHODS: A group of 73 uremic patients in dialysis center of our Department were assigned as uremic group; a group of 30 healthy persons who were examined over the same period were regarded as control group. The level of body mass index (BMI), serum creatinine (SCr), blood urea nitrogen (BUN), leptin, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10, and the neutrophils phagocytosis function were compared in two groups. RESULTS: BMI and IL-10 of uremic group were lower than the control group. The levels of SCr, BUN, leptin, TNF-α, IL-6 of uremic group were higher than the control group (p < 0.05). The neutrophils phagocytosis function in uremic group significantly decreases, compared to control group (p < 0.05). Using one-way ANOVA analysis, serum leptin was positively correlated with the level of TNF-α, IL-6 (r = 0.58, 1.00 respectively, p < 0.05), and was negatively correlated with the level of IL-10 (r = -0.45, p < 0.05). CONCLUSIONS: The high level of leptin and correlated inflammation were involved in the initiation and development of uremia; moreover, leptin was an important mediator.

Effect of nicotine withdrawal on pain sensitivity in rats to mechanical stimulation and thermal stimulation.

OBJECTIVE: To establish an improved rat model of nicotine withdrawal and dependence by subcutaneous injection of pure nicotine, and observe the effect of nicotine withdrawal on the pain sensitivity in rats. MATERIALS AND METHODS: 30 SD rats were randomly divided into 5 groups with 6 rats in each group, including the control group, normal saline group (NS group), nicotine group of 3 mg/kg/d (NT3 group), nicotine group of 9 mg/kg/d (NT9 group) and nicotine group of 18 mg/kg/d (NT18 group). The 5 groups were respectively subcutaneously injected with nothing, normal saline, 1 mg/kg nicotine, 3 mg/kg nicotine and 6 mg/kg nicotine with 3 times per day for 7 consecutive days. 60 min after last injection in the 7th d, 1 mg/kg mecamylamine was subcutaneously injected. The body weight change, survival and nicotine withdrawal score of rats were observed during injection of nicotine and after withdrawal. Mechanical withdrawal threshold (MWT) and Thermal withdrawal latency (TWL) in the right hind sole of another 18 rats selected from the control group, NS group and NT9 group (6 rats from each group) were respectively tested in 7d after injection of normal saline or nicotine. RESULTS: Compared with the NT3 group, the body weight of rats in the NT9 group and NT18 group were slowly increased in 7d after injection of nicotine (p < 0.05), but were rapidly increased in 1d and 2d after withdrawal (p < 0.01). Rats in the NT9 group and NT18 group had more withdrawal symptoms after stimulation with mecamylamine (p < 0.01), but the mortality of rats in the NT18 group reached 17%. Compared with the control group, MWT in the rats of the NT9 group were significantly decreased in 1-7d after nicotine withdrawal (p < 0.01), and were particularly significantly decreased in 1d and 2d (p < 0.01); TWL was also significantly decreased (p < 0.01), and was most significantly decreased in 4d (p < 0.01). CONCLUSIONS: An improved rat model of nicotine dependence and withdrawal can be successfully established by intermittent subcutaneous injection of nicotine at 9 mg/kg/d for 7 days, and the pain sensitivity in rats is increased after nicotine withdrawal.

Primary culture of neurospheres obtained from fetal mouse central nervous system and generation of inner ear hair cell immunophenotypes in vitro.

OBJECTIVE: To investigate whether hair cell immunophenotypes can be derived from the central nervous system. DESIGN: We established in vitro cell cultures from embryonic day 14.5 fetal rat brain tissue, and analysed changes in the immunohistochemical features of these cell cultures following differentiation. RESULT: The immature neural progenitors obtained from the fetal mouse central nervous system generated cell immunophenotypes which expressed epitopes of the hair cell marker proteins myosin VIIa and Brn-3c and the supporting cell marker pan-cytokeratin. CONCLUSION: Neural progenitors have the potential to differentiate into inner ear hair cell and supporting cell phenotypes, and thus may be a useful material for cell transplantation therapy aiming to replace damaged inner ear hair cells.

Insulin-like growth factor-1 suspended in hyaluronan improves cartilage and subchondral cancellous bone repair in osteoarthritis of temporomandibular joint.

This study sought to evaluate the effects of intra-articular injection of insulin-like growth factor-1 (IGF-1) suspended in hyaluronan (HA) on the cartilage and subchondral cancellous bone repair in osteoarthritis (OA) of the temporomandibular joint (TMJ). Disc perforation was performed bilaterally in rabbit TMJs to induce OA. Four groups of animals (n=12) received OA induction only, and either intra-articular HA injection alone, intra-articular IGF-1 injection alone, or a combination of HA and IGF-1 injection. All therapy was begun 4 weeks after OA induction. The animals were killed 12 or 24 weeks after the first injection, for histology and micro-CT examinations. Two additional animals were used as normal controls. Typical cartilage and subchondral cancellous bone lesions were observed in the OA group. No protective effect on cartilage and subchondral cancellous bone was found in the HA or IGF-1 alone groups. Better histological repair and nearly normal micro-architectural properties of the subchondral cancellous bone were observed in the HA+IGF-1 group compared with the HA or IGF-1 alone groups. HA may be used as an effective carrier for intra-articular injection of IGF-1 and the combination of HA/IGF-1 shows promise as a new rational approach to therapy of TMJ OA.

Genetic relationships among Chinese pigs and other pig populations from Hunan Province, China.

In this study, protein-level polymorphisms of transferrin, pre-albumin, hemopexin, ceruloplasmin and amylase were investigated in Hunan native pigs and Large Yorkshire pigs collected from Hunan (a province of China), allowing calculations of allele frequencies, average heterozygosities, inbreeding coefficients and genetic distances. The genetic relationship between Southeast Asian native pigs and American pigs was more distant than those among Southeast Asian native pig breeds. The genetic relationship between Southeast Asian native pig breeds and Hampshire pigs was the most distant.

TIMP-1 regulation of cell cycle in human breast epithelial cells via stabilization of p27(KIP1) protein.

Increasing evidence suggests that tissue inhibitor of metalloproteinases-1 (TIMP-1) can directly regulate cell growth and apoptosis independent of its matrix metalloproteinases (MMPs)-inhibitory activity. While TIMP-1's antiapoptotic activity has been well demonstrated, conflicting data has been reported regarding TIMP-1's role in growth regulation. Here we show that TIMP-1 reduces the growth rate of human breast epithelial (MCF10A) cells by inducing cell cycle arrest at G(1). TIMP-1-mediated cell cycle arrest is associated with its downregulation of cyclin D(1) and upregulation of p27(KIP1), resulting in inhibition of cyclin-dependent kinase activity necessary for phosphorylation of the tumor suppressor retinoblastoma protein. We further show that TIMP-1 modulation of cyclin D(1) and p27(KIP1) is achieved through TIMP-1-mediated differential regulation of protein stability independent of growth factor signaling. We also show that TIMP-1-mediated differential regulation of cyclin D(1) and p27(KIP1) is independent of cell adhesion signaling. Whereas approximately 50% of MCF10A cells with reduced TIMP-1 expression underwent cell death following loss of cell adhesion (anoikis), TIMP-1 overexpressing cells remained viable with prominent cell cycle arrest without detectable cell death. Taken together, we propose that TIMP-1-mediated cell survival independent of cell adhesion is accompanied with cell cycle arrest in human breast epithelial cells, although cell cycle regulation may not be a prerequisite for TIMP-1 regulation of apoptosis in general.