A novel NSUN5/ENO3 pathway promotes the Warburg effect and cell growth in clear cell renal cell carcinoma by 5-methylcytosine-stabilized ENO3 mRNA.

OBJECTIVES: Clear cell renal cell carcinoma (ccRCC) cells often reprogram their metabolisms. Enolase 3 (ENO3) is closely related to the Warburg effect observed in cells during tumor progression. However, the expression and function of ENO3 in ccRCC cells remain unclear. Therefore, this study investigated the expression and functional significance of ENO3 in the Warburg effect observed in ccRCC cells. METHODS: In this study, B-mode and microflow imaging ultrasound examinations were performed to evaluate patients with ccRCC. The extracellular acidification rate test and glucose uptake and lactate production assays were used to examine the Warburg effect in ccRCC cells. Western blotting, quantitative reverse transcription polymerase chain reaction, and immunochemistry were used to detect the expression of ENO3 and NOP2/Sun RNA methyltransferase 5 (NSUN5). RESULTS: ENO3 upregulation in ccRCC tumor tissues was accompanied by an increase in tumor size. Importantly, ENO3 participated in the Warburg effect observed in ccRCC cells, and high levels of ENO3 indicated a poor prognosis for patients. Loss of ENO3 reduced glucose uptake, lactate production, and extracellular acidification rate as well as inhibited ccRCC cell proliferation. Furthermore, NSUN5 was involved in the ENO3-regulated Warburg effect and ccRCC cell progression. Mechanically, NSUN5 was upregulated in ccRCC tissues, and NSUN5 upregulation mediated 5-methylcytosine modification of messenger RNA (mRNA) in ccRCC cells to promote mRNA stability and ENO3 expression. CONCLUSIONS: Collectively, the destruction of the NSUN5/ENO3 axis prevents ccRCC growth in vivo and in vitro, and targeting this pathway may be an effective strategy against ccRCC progression.

Effect of combined treatment including surgery and postoperative adjuvant therapy on spinal metastases of Tomita type 7.

OBJECTIVES: Identify the efficacy of multidisciplinary treatment including palliative spinal surgery on patients with Tomita type 7 spinal metastases. PATIENTS AND METHODS: A retrospective analysis of surgery treated spinal metastatic patients from January 2013 to December 2016 in Tianjin Medical University Cancer Institute and Hospital were performed. Surgical procedures and intraoperative parameters and postoperative adjuvent treatments were studied. Patients' demographic characteristics and medical conditions including paralysis statues, quality of life and pain levels and postoperative survival time were identified. RESULTS: 50 patients were identified with mean age at the time of surgery of 57.68 years old (range 27-78 years). The mean Tokuhashi score was 8.48 and the spinal instability neoplastic score (SINS) averaged at 10.52 points. 48 patients (96%) encountered epidural spinal cord compression. Kaplan-Meier method determined median postoperative survival time was 12.00 months (95% CI: 7.05-16.95 months). The mean score of visual analogue scale (VAS) decreased from 7.66 preoperatively to 1.96 postoperatively. The Frankel scale was improved by at least one grade in 47 patients. Patient's quality of life showed significant improvements. CONCLUSION: Multidisciplinary treatment including palliative spinal surgery was associated with alleviating pain, improving neurologic function and quality of life in patients with Tomita type 7 spinal metastases.

Growth Performance and Meat Quality Evaluations in Three-Way Cross Cattle Developed for the Tibetan Plateau and their Molecular Understanding by Integrative Omics Analysis.

Despite of favorable characteristics of high protein, low fat, and free-pollution, yak meat has intrinsically poor performance in tenderness and color, which is ever challenging yak sector. To this end, a three-way cross system was first developed for high quality beef of the Tibetan Plateau using Angus cattle ( Bos taurus) as terminal sire to mate with 1/2 yak (F1) generated from♂Qaidam cattle ( Bos taurus) × â™€yak ( Bos grunniens). The withers height, chest girth, and body weight of 1/4 yak (F2) were all great higher than that of yak and 1/2 yak ( P < 0.01), especially at later period, suggesting the faster growth rate of 1/4 yak. Also the dressing percentage was much better in 1/4 yak ( P < 0.01). Tenderness and meat color were both significantly improved in 1/4 yak with some unpleasant sacrifice of PUFAs, such as EPA and DHA, and meat protein, given the significantly lower shear force and higher L* ( P < 0.01). A total of 769 genes, including SREBF1, GHR, and FASN, the widely recognized causal genes of meat quality, were identified from 11947 differently expressed genes by the data integration of transcriptome, GWAS and QTL. These genes were significantly enriched for important pathway and GO terms, such as insulin signaling pathway, fatty acid biosynthesis, calcium signaling pathway, metabolic pathway, and cellular response to stress ( P < 0.01). And 12 promising candidates were exemplified with annotation of H3K4me3 data from divergent meat quality, such as OSTF1, NRAS1, and KCNJ11. Interestingly, 75 high-altitude adaptive candidate genes were also detected in the list. This study is a first step toward high quality beef of the Tibetan Plateau and provides useful information for their molecular understanding.

Suppressing Irf2bp2 expressions accelerates metabolic syndrome-associated brain injury and hepatic dyslipidemia.

Increasing studies indicate that high fat diet (HFD) induces neuroinflammation in animal models with obesity, yet the pathology of it is unclear. Interferon Regulatory Factor 2 Binding Protein 2 (Irf2bp2) is a key regulator of macrophage polarization, playing an essential role in regulating inflammatory response. This study investigated the effects of Irf2bp2 on HFD-induced brain injury, and explored the possible molecular mechanisms using wild type (WT) and Irf2bp2 knockout (KO) mice. The results indicated that HFD-increased body weight of mice was further elevated by Irf2bp2-knockout. In addition, Irf2bp2-deletion accelerated HFD-induced metabolic syndrome, as evidenced by the promoted fasting glucose and insulin levels. In the results of behavioral measurements, Irf2bp2-knockout intensified cognitive deficit in HFD-fed mice by using Y-maze, passive avoidance, and morris water maze (MWM) tests. Further, Irf2bp2-deficiency accelerated the activation of astrocytes and microglia cells, as evidenced by the promoted expressions of glial fibrillary acidic protein (GFAP) and Iba-1 in hippocampus and hypothalamus of HFD-fed mice. HFD for 16 weeks induced oxidative stress in serum and brain of mice, as proved by the up-regulated malondialdehyde (MDA) levels and down-regulated superoxide dismutase (SOD) activity, which were significantly enhanced due to Irf2bp2 knockout. Moreover, HFD-triggered systematic and central nervous inflammation by increasing the release of interleukin 1ß (IL-1ß) and tumor necrosis factor (TNF)-α, accompanied with elevated p-nuclear factor-κB (NF-κB) expressions. Notably, HFD-induced inflammation was significantly exacerbated by Irf2bp2 deletion. Intriguingly, HFD-induced dyslipidemia in liver of mice was further aggravated by Irf2bp2 suppression. Our in vitro results verified the effects of Irf2bp2-inhibition on the promotion of inflammatory response in BV2 cells and lipid dysfunction in primary hepatocytes. Therefore, the findings above suggested that inhibiting Irf2bp2 expression provided a potential therapeutic approach for the prevention of metabolic syndrome-associated central nervous injury.

Integrating CNVs into meta-QTL identified GBP4 as positional candidate for adult cattle stature.

Copy number variation (CNV) of DNA sequences, functionally significant but yet fully ascertained, is believed to confer considerable increments in unexplained heritability of quantitative traits. Identification of phenotype-associated CNVs (paCNVs) therefore is a pressing need in CNV studies to speed up their exploitation in cattle breeding programs. Here, we provided a new avenue to achieve this goal that is to project the published CNV data onto meta-quantitative trait loci (meta-QTL) map which connects causal genes with phenotypes. Any CNVs overlapping meta-QTL therefore will be potential paCNVs. This study reported potential paCNVs in Bos taurus autosome 3 (BTA3). Notably, overview indexes and CNVs both highlighted a narrower region (BTA3 54,500,000-55,000,000 bp, named BTA3_INQTL_6) within one constructed meta-QTL. Then, we ascertained guanylate-binding protein 4 (GBP4) among the nine positional candidate genes was significantly associated with adult cattle stature, including body weight (BW, P < 0.05) and withers height (WHT, P < 0.05), fitting GBP4 CNV either with three levels or with six levels in the model. Although higher copy number downregulated the mRNA levels of GBP2 (P < 0.05) and GBP4 (P < 0.05) in 1-Mb window (54.0-55.0 Mb) in muscle and adipose, additional analyses will be needed to clarify the causality behind the ascertained association.

Strontium ranelate prevents bone loss in a rat model of localized muscle paralysis.

Twenty-one 3.5-month-old female Sprague-Dawley rats were randomly assigned to three groups: BTX group, in which each rat received a single intramuscular injection of 2 U of Clostridium botulinum toxin (BTX) in the quadriceps femoris muscle of the right hind limb; BTX + SR group, in which each rat received a BTX injection and a dose of strontium ranelate (dose level of 625 mg/kg/day); and the control group. All the rats were killed at 9 weeks post-treatment. It was showed that BTX-induced rats a rapid loss of body weight in the first 3 weeks, after which their body weight showed a slow increase similar to that observed in the control rats. The net body weight loss was mainly attributed to muscle atrophy. BTX caused remarkable bone degradation in either the trabecular bone or the cortical bone of the disuse femur. The deteriorations in the bone mass and bone microstructure were locally limited and could be prevented by strontium ranelate treatment. Biomechanical analysis showed that strontium ranelate treatment improved the mechanical performance of the tibia in BTX-treated rats. It was showed that a clinical-corresponding dose of strontium ranelate could prevent bone loss in long-term immobilized rats.

Omentin inhibits osteoblastic differentiation of calcifying vascular smooth muscle cells through the PI3K/Akt pathway.

Arterial calcification is positively associated with visceral adiposity, but the mechanisms remain unclear. Omentin is a novel adipokine that is selectively expressed in visceral adipose tissue. The levels of circulating omentin are decreased in obesity, and they correlate negatively with waist circumference. This study investigated the effects of omentin on the osteoblastic differentiation of calcifying vascular smooth muscle cells (CVSMCs), a subpopulation of aortic smooth muscle cells putatively involved in vascular calcification. Omentin inhibited mRNA expression of alkaline phosphatase (ALP) and osteocalcin; omentin also suppressed ALP activity, osteocalcin protein production, and the matrix mineralization. Furthermore, omentin selectively activated phosphatidylinositol 3-kinase (PI3K) downstream effector Akt. Moreover, inhibition of PI3K or Akt activation reversed the effects of omentin on ALP activity and the matrix mineralization. The present results demonstrate for the first time that omentin can inhibit osteoblastic differentiation of CVSMCs via PI3K/Akt signaling pathway, suggesting that the lower omentin levels in obese (specially visceral obese) subjects contribute to the development of arterial calcification, and omentin plays a protective role against arterial calcification.

Glucose metabolism disorders in cancer patients in a Chinese population.

BACKGROUND: Characteristics of glucose metabolism disorders (GMDs) in different cancers and the contributory role of GMDs in developing cancers are still not so clear. METHODS: Two thousand four hundred and five patients with malignancy who had been hospitalized in the First Affiliated Hospital of Jinan University were pooled as case group. Two thousand and sixteen non-cancer people who finished health examinations in the Affiliated Yangcheng Hospital of Guangzhou Medical College were enrolled as control group. We compared glucose metabolism among patients with different kinds of malignancy. Based on logistic regression models, we analyzed factors that affect the development of carcinoma. RESULTS: (1) Among 2,408 malignancy patients, the total prevalence of diabetes mellitus (DM) and impaired fasting glucose (IFG) reached 28.0%. Pancreatic cancer, lymphoma, liver cancer, leukemia, and colorectal cancer showed most striking hyperglycemia. (2) Leukemia and esophageal cancer accounting for 12.5% and 12.1%, respectively, were the most likely to suffer from hypoglycemia. (3) Older cancer patients seem to be more vulnerable to hyperglycemia, while the younger tend to be more likely to develop hypoglycemia. (4) High level of fasting plasma glucose (FPG) was associated with lung cancer, breast cancer, leukemia, lymphoma, thyroid cancer, bladder cancer, and pancreatic cancer. Patients with DM increased risks for developing colorectal cancer, liver cancer, esophageal cancer, thyroid cancer, cervical cancer, and pancreatic cancer. CONCLUSIONS: GMDs are frequent events in malignancy patients. Hyperglycemia and hypoglycemia are found in the same kinds or different kinds of cancers, and the incidence of hyperglycemia is higher than that of hypoglycemia. Characteristics of GMDs were dissimilar in different cancers and different ages. Hyperglycemia was a risk factor for many cancers.

[Surgical treatment and prognosis of cancer of hepatic flexure of colon invading the duodenum in 65 patients].

OBJECTIVE: To discuss surgical treatment of right colon carcinoma of hepatic flexure invading the duodenum. METHODS: Sixty-five patients with right colon carcinoma of hepatic flexure invading the duodenum, treated in our department from 1987 to 2007, were included in this study. Their clinicopathological data were retrospectively reviewed and analyzed. All the cases were divided into three types (local invasion, regional invasion, and cancer with internal fistula) according to duodenal defect, including local invasion (< 2.0 cm), wide invasion (> 2.0 cm) and the presence of internal fistula. RESULTS: 25 patients with local invasion underwent en bloc resection of the duodenal wall. Pedicled ileal flap was used to cover the large duodenal defect measuring 2.0 - 3.0 cm in 5 patients. Dudenojejunostomy was used to reconstruct the large defect measuring more than 5 cm in 3 patients. Conservative resection of right-sided colon was performed in 18 patients with wide invasion. 4 patients underwent pancreaticoduodenectomy combined with right hemicolectomy for colon cancer involving the pancreatic head. 10 underwent duodenal diverticularization. One patient with anastomotic leakage healed within 3 weeks. Other patients were cured without postoperative complications. The total 3-year and 5-year survival rates after surgery were 53.8% and 9.2%, respectively. CONCLUSION: The surgical procedure to be performed is usually decided according to the cancer location, extent, and duodenal defect and invasion, which are important for prolonging life time, improving of quality of life and prognosis in these patients.

[Dynamic effects of glucocorticoid on bone mineral density and microarchitecture: experiment with rats].

OBJECTIVE: To investigate the Dynamic effects of glucocorticoid (GC) on bone mineral density and microarchitecture time-related changes of trabecular bone in bone mineral density (BMD) and microarchitecture in glucocorticoid-treated rats. METHODS: Fifty-two 3.5-month-old female SD rats were randomly divided into 3 groups. Ten rats were killed at the beginning of experiment with their right tibiae taken out as the baseline group; 22 rats underwent subcutaneous injection of methylprednisolone once daily (GC-treated group), and the other 20 rats underwent subcutaneous injection of normal saline once daily as control group. One and 9 weeks after the beginning of experiment 11 and 10 rats from GCT Group and control group each were killed with their right tibiae taken out. High resolution micro-CT was used to identify the densitometric and microarchitectural properties of the trabecula in the proximal metaphysic of tibia. RESULTS: Compared with the control group the values of volumetric BMD (vBMD), tissue BMD (tBMD), bone volume fraction (BVF), trabecular number (Tb.N), degree of anisotropy (DA), and trabecular connectivity (Conn.D) in the trabecular bone at different time-points, of the GCT group all decreased; and the values in the ninth week were the lowest (all P < 0.05). The values of trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), and structure model index (SMI) at different time-points of the GCT group were higher than those of the control group. A time-related analysis within the GCT group showed there was a declination in BVF, Conn.D, Tb.N, and DA with administration time, but Tb.Th and Tb.Sp were increased significantly (all P < 0.05). The mean values of Tb.Th in the first week and the ninth week of GCT Group were (0.076 +/- 0.020) mm and (0.086 +/- 0.026) mm respectively, both higher than the baseline value [(0.067 +/- 0.014) mm] and the values of the control group in the first and ninth weeks [(0.075 +/- 0.022) mm and (0.072 +/- 0.009) mm respectively]. CONCLUSION: Administration of GC time dependently decreases the BMD and causes deterioration in microarchitecture of trabecular bone; and the remaining trabeculae seem thicken to increase their strength as compensation.

[Caudal-related homeobox 2 gene expression and its effects on human gastric carcinoma cell line].

OBJECTIVE: To investigate the effect of human caudal-related homeobox 2 (Cdx2) gene expression on human gastric carcinoma cell line MGC-803. METHODS: pCMV-Cdx2-HA eukaryotic expression plasmid was constructed by using DNA recombinant method. The MGC-803 cells were divided into 4 groups: non-transfected, transfected with pCMV-HA, transfected with pCMV-GAPDH-HA, transfected with pCMV-Cdx2-HA. Cdx2 gene and its protein expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot techniques respectively. The effects of Cdx2 overexpression on the growth of MGC-803 cells in vitro was assessed by measuring MTT, flow cytometry and transmission electron microscopy analysis. RESULTS: In MGC-803 cells transfected with pCMV-Cdx2-HA, cell proliferation was significantly suppressed, the cells was ultrastructurally destroyed, cell cycle progression was blocked in G0/G1 and apoptosis rate was higher (P<0.05) in comparison with non-transfected cells or cells transfected with empty vector. CONCLUSION: It indicated that Cdx2 gene can suppress the growth of gastric carcinoma and may save as a novel therapeutic target.

The influence of center of rotation on the assessment of trabecular bone densitometric and structural properties.

The center of rotation is a physical location in the microCT scanner, defined by the axis of rotation of the sample stage. This physical location is always well defined during calibration of the instrument and fitted by an appropriate algorithm. However, in real images of limited contrast and with X-ray photon noise, this algorithm exhibits poorer precision and the optimum center of rotation cannot be always acquired. Thus, adjustment by operator is necessary to determine whether the center of rotation was correct, in order that the structural information of the sample can be correctly interpreted. In this paper, the effect of center of rotation on the assessment of densitometric and structural properties of trabecular bone was firstly evaluated. Twenty female Sprague-Dawley rats of 7-month-old were randomly assigned to ovariectomized (OVX) and SHAM-operated (SHAM) groups. The left tibiae were harvested at 3 weeks postoperatively. High resolution microCT was used to identify the densitometric and microstructural properties of trabeculae in the proximal ends of tibia. After CT scanning, the best artificial center of rotation for each scan was obtained. Bone parameters analyses were performed on the centers at different places away from the best artificial center of +/-0.2, +/-0.5, +/-1.0, +/-1.5, and +/-2.0 pixels, respectively. The general linear model (GLM) repeated measures procedure was used to investigate the difference in the parameters between the two groups (OVX vs. SHAM) and the possible effects of center displacements. A significant difference between OVX and SHAM groups was found in all parameters (p < 0.05) except Tb.Th, DA, and BS/BV. TBMD, DA, BS/BV, and Conn.D were decreased while BV/TV and Tb.Th were increased with the center deflection. Variations of these parameters were acceptable when the displacements were limited within +/-1.5 pixels for tBMD, BV/TV, DA, and Conn.D, and +/-1.0 pixels for Tb.Th and BS/BV. These changes were similar in both OVX and SHAM groups. The changing curves of bone parameters vs. centers could be well fitted by quadric regression models, by which the real center could be acquired, and thus the precision of microCT analysis would be improved. There were some inevitable differences between the best artificial and real centers.

[Inhibitory effects of transcription factor E2F-1 siRNA on invasion and proliferation of gastric cancer cell line MGC803].

BACKGROUND & OBJECTIVE: As an important regulatory factor of cell cycle, transcription factor E2F-1 is closely related to tumorigenesis. This study was to investigate the effects of E2F-1 small interfering RNA (siRNA) on invasion and proliferation of human gastric cancer MGC803 cells. METHODS: E2F-1 siRNA vector containing short hairpin structure was transfected into MGC803 cells. Untransfected and pSilencer4.1-negative-transfected cells were used as controls. The expression of E2F-1 was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Cell matrigel invasion assay and cloning assay were used to detect the invasion and proliferation of MGC803 cells after E2F-1 siRNA transfection. RESULTS: At 48 h after E2F-1 siRNA transfection, the mRNA level of E2F-1 was down-regulated by over 90.0% of controls; the protein level of E2F-1 was down-regulated by 79.6% of negative control and by 81.5% of empty control. The number of migrated cells was significantly smaller in E2F-1 siRNA group than in negative and empty control groups (18.0+/-2.6 vs. 48.0+/-4.6 and 54.0+/-5.6, P<0.05). The number of cell clones was also significantly smaller in E2F-1 siRNA group than in negative and empty control groups (46.0+/-2.0 vs. 122.3+/-1.5 and 128.7+/-2.1, P<0.05) CONCLUSION: E2F-1 siRNA could down-regulate E2F-1 expression in human gastric cancer MGC803 cells and suppress its invasion and proliferation to some extent.

Quantitative associations between osteocyte density and biomechanics, microcrack and microstructure in OVX rats vertebral trabeculae.

Osteocytes actively regulate bone modeling and remodeling, direct skeletal mineralization, and regulate calcium/phosphate homeostasis and extracellular matrix metabolism; yet the specific role of osteocytes in maintaining bone structural integrity and strength is unknown. Studies have shown that the density of osteocytes decreases with age and estrogen deficiency, as seen in postmenopausal women. Here, we examined the relationships between osteocyte density and the related variables, including biomechanics, bone mineral density, microcrack and microstructure of vertebral trabeculae, in ovariectomized rats. We found that osteocyte density correlated with some of the parameters that determine the biomechanical quality of bone. Our findings suggest that osteocytes could play a crucial role in maintaining the mechanical quality of bone, and osteocyte density could be considered as an alternative index in assessing bone quality.

[Clinical analysis of gastroparesis syndrome after nongastrectomy abdominal operation: a report of 22 cases].

OBJECTIVE: To investigate the causes, diagnosis and treatment of gastroparesis syndrome after nongastrectomy abdominal operation. METHODS: The clinical data of 22 cases with gastroparesis syndrome after nongastrectomy abdominal operation from 1972 to 2004 were retrospectively analyzed. RESULTS: Gastroparesis syndrome after nongastrectomy abdominal operation often occurred during 4-6 days postoperatively when the patients began to take in food, characterized by upper abdominal distension, nausea, vomiting, strong splashing bowel sound, weak bowel sound and large quantity of gastric drainage ranging from 1000 to 3000 ml every day. Barium meal was valuable not only in the diagnosis but also effective for promoting gas motility. It revealed a non-peristaltic, flabby and static stomach, and retention of contrast medium in the stomach even 5-6 hours later. All the patients recovered through non-operative therapy for 5-25 days including continuous gastrointestinal decompression, TPN and gastro-intestinal dynamic medicine. CONCLUSIONS: The causes of gastroparesis syndrome after nongastrectomy abdominal operation are multifactorial, most of such patients can be cured by non-operative therapy.

[Factors influencing intelligence in elderly population in a community-based study in Taiyuan city].

OBJECTIVE: To explore the factors which influencing the intelligence in elderly in a community, so as to provide reference on primary prevention of dementia. METHODS: A door to door survey was conducted. A total number of 830 elderly were assessed using WAIS-RC, H-NTLA and data collected through questionnaires on lifestyles, family and social activity were analysed, using the factor analysis, stepwise regression and canonical correlation analysis. RESULTS: The main variables related to the verbal were eggs diet, blood pressure, fruit, visit their children, alcohol intake (standard coefficients of regression are -0.118, -0.079, -0.060, -0.036, -0.117, respectively). The main variables related to cognition were recreation, sports and hereditary history of mental disorders (standard coefficients of regression were 0.035, -0.127, respectively). The main variables related to memory were contact to chemical materials, age of their parents during delivery, alcohol intake and the relationship between husband and wife (standard coefficients of regression were -0.063, 0.055, -0.030, -0.037, respectively). The variables related to canonical variable V(1) would include education and occupation (canonical correlation = 0.5993, P = 0.0001) while V(2) would include cerebrovascular accident (canonical correlation = 0.3925, P = 0.0005). CONCLUSION: Intelligent work, family harmony, prevention of cerebrovascular diseases were the main areas to prevent intellectual disability in the elderly.