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Objectives: To investigate the protective effect and mechanism of total flavonoids from Citrus paradise cv. Changshan-huyou extracts (TFC) on oxygen-glucose deprivation (OGD) of primary neurons and chronic ischemia-induced cerebral injury in mice., Methods: Primary hippocampal neurons of 18-day fetal rats were isolated and cultured for 1 week, then treated with 0.25, 0.50 and 1.00 mg/mL TFC. After oxygen-glucose deprivation for 1 h, cells were reperfused for 6 and 24 h, respectively. The cytoskeleton was observed by phalloidin staining. In animal study, 6-week ICR male mice were randomly divided into sham operation group, model group, low-dose (10 mg/kg), medium-dose (25 mg/kg) and high-dose (50 mg/kg) TFC treatment groups, with 20 mice in each group. After 3 weeks, chronic cerebral ischemia was induced by unilateral common carotid artery ligation in all groups except sham operation group. Mice were treated with different concentrations of TFC in the three TFC treatment groups for 4 weeks. Open field test, novel object recognition test and Morris water maze test were used to evaluate anxiety, learning and memory of these mice. Nissl, HE and Golgi stainings were used to detect neuronal degeneration and dendritic spine changes in the cortex and the hippocampus. The expression levels of Rho-associated kinase (ROCK) 2, LIM kinase (LIMK) 1, cofilin and its phosphorylation, as well as the expression of globular actin (G-actin) and filamentous actin (F-actin) protein in hippocampus of mice were detected by Western blotting., Results: Neurons subjected to OGD showed that neurites displayed shortening and breakage; while treatment with TFC reversed OGD-induced neurite injury, especially in the 0.50 mg/mL TFC group. Compared with the sham operation group, the mice in the model group showed a significant decline in anxiety and cognitive ability ( P <0.01), whereas treatment with TFC significantly reversed anxiety and cognitive deficits ( P <0.05). Improvement in the medium-dose TFC group was the most obvious. Histopathological analysis indicated that the number of Nissl bodies and dendritic spines in hippocampus and cortex were decreased in the model group (all P <0.01). However, after treatment with medium dose of TFC, the number of Nissl bodies and dendritic spines (all P <0.05) was significantly recovered. Compared with the sham operation group, the phosphorylation level of ROCK2 in the brain tissue of the model group was significantly increased ( P <0.05), while the phosphorylation levels of LIMK1 and cofilin were significantly decreased ( P <0.05), and the relative content ratio of G-actin/F-actin was significantly increased ( P <0.05). After administration of TFC, the phosphorylation level of ROCK2 in brain tissue of each group was significantly decreased ( P <0.05), while the phosphorylation levels of LIMK1 and cofilin were significantly up-regulated ( P <0.05) and the relative content ratio of G-actin/F-actin was significantly decreased ( P <0.05)., Conclusions: TFC protects from ischemia-induced cytoskeletal damage, reduces neuronal dendritic spine injury and protects mice against chronic cerebral ischemia through RhoA-ROCK2 signaling pathway, indicating that TFC might be a potential candidate for treatment of chronic ischemic cerebral injury.

Objective: To explore the impact of dietary sodium-intake on residual renal function in patients undergoing peritoneal dialysis (PD)., Methods: Thirty-three patients on PD with stable dialysis were regularly followed up for 12 months. The daily sodium intake of the patients was calculated based on the 3-day dietary record. Based on the mean daily sodium intake, the patients enrolled were divided into low-salt group (sodium intake≤3.0 g/day, 19 patients) and high-salt group (sodium intake&gt;3.0 g/day, 14 patients). The baseline data of the patients were recorded, and the indicators of residual renal function and peritoneal function were regularly tested. The patients were followed-up at 3-month intervals, and their urine volume, peritoneal ultrafiltration volume and other clinical indicators were recorded and the biochemical indexes were detected to evaluate the changes in the residual renal function and peritoneal function., Results: There was a positive correlation between the total sodium excretion and dietary sodium intake in these patients ( r =0.536, P =0.0013), and sodium excretion by dialysis was positively correlated with their sodium intake ( r =0.901, P =0.000). Regression analysis suggested that the total sodium excretion was correlated with dietary sodium intake (β=0.416, 95% CI : 0.170-0.666; P &lt; 0.0018); sodium excretion by dialysis was associated with dietary sodium intake (β=0.489, 95% CI : 0.395-0.582; P &lt; 0.001). The residual renal function was reduced by 17.48±11.22 L /(w·1.73 m 2 ) in the low-salt group, as compared to 30.20±18.30 L /(w·1.73 m 2 ) in the high-salt group ( P =0.032). The reduction in the residual renal function was correlated with sodium intake in the PD patients ( r =0.409, P =0.018). Multivariate regression analysis showed that sodium intake was an independent factor contributing to the reduction of residual renal function (β=14.646, 95% CI 7.426-21.866, P &lt; 0.001)., Conclusions: Sodium excretion by PD in patients with continuous ambulatory PD is positively correlated with their dietary sodium intake, which contribute to the decrease of residual renal function. A high dietary sodium intake may accelerate the reduction of residual renal function in these patients.

Objective To study the effect of muscle segment homeodomain homeobox 2 (MSX2) on the expression of enamel matrix protein and the formation of enamel. Methods Immunohistochemical staining was used to detect the expression of MSX2 in mouse tooth embryos and its localization in ameloblasts. The short hairpin RNA (shRNA) of the MSX2 gene was designed and synthesized, and then the annealed double stranded DNA was constructed into the pGMLV-SC5 RNAi lentivirus vector, and finally it was packaged with lentivirus. The lentivirus was used to infect ameloblasts. Real-time fluorescent quantitative PCR was performed to screen the best interference fragment, and detect the mRNAs of amelogenin (Amelx), ameloblastin (Ambn), enamelin (Enam), amelotin (Amtn) and kallikrein 4 (Klk4). The embryos were isolated for 18.5 days and then infected with RNAi recombinant lentivirus targeting MSX2. The tooth germ was implanted under the renal capsule of the mouse. Ten weeks later, the tissue was harvested to separate and observe the tooth form and contour. Results MSX2 was expressed in the secretory phase and maturation phase of mouse ameloblasts, but the expression signal was weaker in the secretory phase and was stronger in the mature stage. The lentivirus of MSX2-shRNA targeting MSX2 gene we constructed inhibited the expression of Amelx and Klk4 mRNAs. The RNAi lentivirus targeting MSX2 gene infected the tooth enamel and led to a decrease in the degree of enamel mineralization. Conclusion The MSX2 gene is expressed in ameloblasts. The knockdown of MSX2 can inhibit the expression of enamel matrix proteins and the enamel mineralization.

Objective: To evaluate the association between central pulse pressure level and cardiovascular diseases (CVD) in patients receiving maintenance peritoneal dialysis., Methods: This cross-sectional study was conducted in 234 patients on maintenance peritoneal dialysis. Central pulse pressure levels were measured using a SphygmoCor analyzer, and cardiovascular diseases were defined as diagnosed ischemic heart disease, heart failure, stroke or peripheral vascular disease., Results: The central pulse pressure levels of patients with CVD were significantly higher than those without CVD (51.7 ± 22.5 vs 43.7 ± 17.8 mmHg, P=0.004), while the brachial pulse pressure levels were comparable between the two patient groups. After adjusting for brachial pulse pressure and other relevant risk factors, central pulse pressure level was found to independently associate with CVD (adjusted OR=1.33, 95% CI 1.01-1.73, P=0.04 )., Conclusion: High central pulse pressure level may serve as a risk factor for CVD in patients on maintenance peritoneal dialysis.

Objective: To evaluate the quality of Rhizome of D. persimilis Prain et Burk., Methods: Determined the contents of 6 qualitative indexes in samples of Rhizome of D. persimilis Prain et Burk that were cultivated in Yongning, Bobai, Beiliu, Guiping and Rongxian of Guangxi and in sample of Rhizome of Dioscorea opposita Thunb, that was cultivated in Wenxian, Henan province., Results: The contents of water in them (%): 15.39, 18.52, 15.41, 13.96, 15.48, 15.74; the contents of protein in them (%): 7.06, 8.45, 9.20, 9.67, 8.63, 9.79; the contents of starch in them (%): 84.51, 86.51, 82.72, 83.65, 84.19, 79.13; the contents of water extracts in them (%): 6.19, 6.98, 8.95, 7.41, 6.70, 9.40; 90% EtOH extracts in them (%): 2.00, 2.04, 2.68, 2.20, 2.11, 3.53; the contents of 18 kinds of amino acids in them (%): 5.23, 6.31, 6.22, 6.78, 6.59, 6.83., Conclusion: The quality of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu is beter than that of Rhizome of D. persimilis Prain et Burk cultivated in Yongning. The quality of Rhizome of D. persimilis Prain et Burk cultivated in Rongxian or Bobai are between those of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu and Rhizome of D. persimilis Prain et Burk cultivated in Yonging. The qualities of Rhizome of D. persimilis Prain et Burk cultivated in Guiping or Beiliu are similar to that of Yam.