Objective To investigate sequential expression of bone sialoprotein(BSP), alkaline phosphatase(ALP) and cementum attachment protein(CAP) in cementoblast(CB) during mineralized culture in vitro, and study the morphological and biologic characters of the CB in this process. Methods CB was seeded on the glass coverslips, and cultured for 6h, 12h, 1d, 2d, 3d, 4d, 5d, 6d, 8d, 10d, 12d and 16d, respectively. The expression of BSP, ALP and CAP proteins was detected using immunocytochemical method. Results 6 hours after plated, cementoblast expressed all of the three proteins. In the second and third days after plated, the cells became confluent and formed multiple layers, BSP and ALP expression decreased, and CAP did not be expressed at all. From the forth day, the cells formed cell nodules with mineralizing function. The cells in the nodules strongly expressed BSP,ALP and CAP, the cells around the nodule weakly expressed BSP and ALP, and did not express ALP. During the following 10 to 16 days, the cell nodules became mineralized nodules. Conclusion These results elucidated the changes of ALP, BSP and CAP expression as well as cell morphology during the CB proliferation, differentiation and mineralization in vitro, and provided some valuable information for studying the formation of cementum and CB proliferation, differentiation and mineralization in vivo.

Objective To study the ultrastrueture of 3 types of periodontal cells, including periodontal ligament cells (PDLC), os-teoblast cells (OB) and eementoblast cells (CB). Methods After culturing 3 kinds of periodontal cells - PDLC, OB and CB, we observed them with transmission electron microscope. Results There was rich rough endoplasmic reticulum and lots of microfilaments in the cyto-plasm of CB and PDLC cells. There was rich rough endoplasmic in the cytoplasm of OB cells. Conclusion The main characteristic ultra-structure feature of the bovine CB and PDLC was rich rough endoplasmic reticulum and microfilaments in the cytoplasm. Compared with CB and PDLC, OB contained fewer microfilaments in the cytoplasm.

Objective To analyze the effects of individualized lifestyle intervention on compliance and metabolic status of patients with type 2 diabetes mellitus (T2DM).Methods Two hundred T2DM patients were selected and randomly divided into experimental and control groups of 100 patients respectively.The experimental group was given individualized lifestyle intervention for 6 months in addition to conventional oral medications.The intervention was to prescribe diet control and exercise therapy according to the patients' individual conditions.The control group was given conventional treatment and verbal lifestyle intervention for 6 months.Comparison was made in patients compliance and various metabolic markers between the two groups.Results The percentage of conduction of diet control and exercise therapy in experimental group was significantly higher than control group ( Diet control:80 vs.52,x2=7.08,P=0.029;Exercise therapy:78 vs.44,x2=11.207,P=0.004).After intervention,the fasting plasma glucose (FPG),2-hour postprandial blood glucose (2hPG),glycated hemoglobin ( HbA1c),body mass index ( BMI),triglyceride ( TG),total cholesterol (TC),low-density lipoprotein ( LDL-C ),and insulin resistance index ( HOMA-IR ) in experimental group decreased significantly,and high-density lipoprotein ( HDL-C ) increased significantly [FPG:( 8.45 ± 1.46 ) mmol/L vs.(6.66 ± 0.67) mmol/L,P=0.000;2hPG:( 12.76 ± 2.25 ) mmol/L vs.(8.22 ± 1.79) mmol/L,P=0.000;HbA1c:(7.68 ± 1.06 ) ％ vs.( 6.48 ± 0.69 ) ％,P=0.000;BMI:( 25.90 ± 1.72 ) kg/m2 vs.( 22.81 ±1.41 ) kg/m2,P=0.016;TG:(2.57 ±0.68) mmol/Lvs.( 1.88 ±0.35) mmol/L,P=0.006;TC:(5.72 ±0.13) mmol/L vs.(5.14 ± 1.38) mmol/L,P=0.043;LDL-C:(3.28 ±0.10)mmol/L vs.(2.81 ±0.57)mmol/L,P=0.009;HOMA-IR:7.58 ± 0.19 vs.4.58 ± 1.98,P=0.000;HDL-C:( 1.29 ± 0.04) mmol/L vs.( 1.62 ± 0.27 ) mmol/L,P=0.003].The levels of FPG,2hPG,HbA1c,BMI,TG,HOMA-IR also decreased in control group after intervention compared with before intervention [FPG:( 8.67 ± 2.71 ) mmol/L vs.( 7.26 ± 1.21 ) mmol/L,P=0.001;2hPG:( 12.82 ± 2.15 ) mmol/L vs.( 10.85 ± 1.98 ) mmol/L,P=0.000,HbA1c:( 7.75 ± 1.08 ) ％ vs.( 7.01 ± 0.87 ) ％,P=0.002;BMI:( 25.82 ± 1.74 ) kg/m2 vs.( 24.23 ± 1.36 ) kg/m2,P=0.024;TG:(2.47 ±0.75) mmol/L vs.(2.13 ± 0.43 ) mmol/L,P=0.018;HoMA-IR:7.88 ± 0.20 vs.6.15 ± 2.01,P=0.042].No significant difference was found on the values of TC,HDL-C and LDL-C before and after intervention in control group (P ＞ 0.05).The effect of intervention of experimental group was more obvious when compared with control group ( FPG:P=0.036;2hPG:P=0.000;HbA1c:P=0.045;BMI:P=0.037;TG:P=0.022;HoMA-IR:P=0.000).Conclusion Individualized lifestyle intervention can improve the compliance of T2DM patients,and was in favor of control metabolic status of T2DM patients to delay the occurrence and development of complications.

Objective To investigate the mechanisms of nitric oxide (NO) in decreasing intestinal mesenteric arterial hypocontractility in rats with hepatic cirrhosis and portal hypertension,and to analyze the interaction of NO and RhoA/ROCK pathway.Methods The levels of NO in the peripheral blood and mesenteric artery of normal rats (normal control group,5 rats),rats with portal hypertension (experimental control group,6 rats)and rats with portal hypertension treated by L-NAME (L-NAME group,6 rats) were detected.Mesenteric arteriole contractility to norepinephrine in the 3 groups was determined using a vessel perfusion system.The expressions of proteins of NO-cGMP-PKG pathway and RhoA/ROCK pathway in the 3 groups were detected by Western blot.All data were analyzed using the analysis of variance or LSD-t test.The changes of mesenteric arteriole contractility to norepinephrine was expressed in dose-response curve,and was analyzed using the nonlinear regression method,and the EC50 value was calculated.Results (1) The pressures of portal veins of the normal control group,experimental control group and L-NAME group were (6.2 ± 0.9)mm Hg (1 mm Hg =0.133 kPa),(13.9 ± 1.7)mm Hg and (16.6 ± 1.3) mm Hg,respectively,with a significant difference among the 3 groups (F =94.4,P ＜ 0.05).(2) The levels of NO in the normal control group,experimental control group and L-NAME group were (43 ± 5) μmol/L,(82 ± 16) μmol/L and (45 ± 9) μmol/L,respectively,with a significant difference among the 3 groups (F =24.77,P ＜ 0.05).The level of NO of the L-NAME group was significantly lower than that of the experimental control group (P ＜ 0.05).(3) The levels of NO in the mesenteric artery of the normal control group,experimental control group and L-NAME group were (236 ±41) μmol/g,(407 ± 82) μmol/g and (216 ± 42) μmol/g,respectively,with a significant difference among the 3 groups (F =20.29,P ＜ 0.05).The NO level of the L-NAME group was significantly lower than that of the experimental control group (P ＜ 0.05).(4) Compared with the experimental control group,the dose-response curve of mesenteric arterioles to norepinephrine shifted to the left,while it did not reach the level of the normal control group.The levels of EC50 of the normal control group,experimental control group and the L-NAME group were 6.458 × 10-7 mol,9.546 × 10-7 mol and 7.494 × 10-7 mol,respectively.There was a significant difference in the EC50 level between the L-NAME group and the other two group (t =2.726,3.112,P ＜ 0.05).(5) Compared with the normal control group,the protein expression levels of eNOS and p-VASP of mesenteric artery of the experimental control group were significantly increased (P ＜ 0.05),while they were decreased in the L-NAME group (P ＜ 0.05).The protein expression levels of eNOS and p-VASP of mesenteric artery of the L-NAME group were significantly higher than those of the normal control group (P ＜0.05).There were no obvious changes of protein expression levels of PKG-1,ROCK-1 and p-moesin in the 3 groups (P ＞ 0.05).(6) The activity of ROCK-1 was significantly increased with norepinephrine stimulation in the normal control group and the L-NAME group,while no obvious changes were detected in the experimental control group.Conclusions The NO expression is upregulated in mesenteric arteries in rats with hepatic cirrhosis and portal hypertension.Such changes induce ROCK activation via influencing the expression of vasoconstrictors.L-NAME can reduce the NO levels in the mesenteric arteries,which may improve RhoA/ROCK signal pathway transduction.This can help vasoconstrictors induce ROCK activation without affecting the protein expression of ROCK.

OBJECTIVETo isolate and culture bovine cementum-derived cells and investigate their biological properties.

METHODSCulture of primary bovine cementoblast (CB) were established from new-born bovine teeth. Cementum was manually dissected, fragmented, and digested twice with collagenase. Following a thorough wash to remove liberated cells, the remaining cementum fragments were plated and cultured in RPMI1640 containing 10% FCS and 1.5% Hepes. The cells in culture were identified using immunocytochemistry by expression of cementum attachment protein (CAP), osteocalcin (OCN) and alkaline phosphatase (ALP). ALP activity was measured by modified Gomori staining.

RESULTS(1) The cells in culture possessed the typical morphology of CB and there were no obvious change of the cell morphology up to 5th passage; (2) Cells in culture exhibited alkaline phosphatase activity and were also positive to CAP, OPN and ALP immunohistochemistry staining.

CONCLUSIONThe cells isolated and cultured in this experiment possess the properties of CB. This enables us to further observe its biological characteristics during cementogenesis and periodontal regeneration.

Alkaline Phosphatase ; metabolism ; Animals ; Cattle ; Cell Separation ; methods ; Cells, Cultured ; Dental Cementum ; cytology ; enzymology ; Immunohistochemistry

Objective: To compare the short-term outcomes of a collagen matrix (CM) and free gingival graft (FGG) in augmenting keratinized mucosa around dental implants. Methods: Nineteen partially edentulous patients who had undergone implant surgery or implant review from June 2017 to June 2018 at Department of Periodontology, Peking University School and Hospital of Stomatology with lack of keratinized mucosa at buccal aspect of implants (<2 mm) were recruited in this study. According to the width of keratinized mucosa (KW) pre-operation, 9 patients including 5 males and 4 females were assigned into control group (KW<0.5 mm) which performed free gingival graft (17 implants) and 10 patients including 3 males and 7 females were assigned into experimental group (KW≥0.5 mm) which used collagen matrix as the grafts (15 implants). The KW at buccal aspect of each implant were measured pre-operation and 2 weeks, 1 month, 2 months, 3 months after surgery respectively. Each of the patients was required to fill out a questionnaire using a visual analogue scale to assess the postoperative morbidity. Results: The KWs around implants were increased significantly during the 3 months follow-up period in both groups (P<0.01). At 3 months after surgery, KW gain in control group was (3.44±1.64) mm, in experimental group was (2.30±0.82) mm, the difference between two groups was statistically significant (P<0.05). Meantime, the total shrinkage of KW in control group [(34±25)%] and experimental group [(51±11)%] also showed a statistically significant difference (P<0.01). However, by using collagen matrix as the grafts, augmented tissues had a much more comparable appearance with adjacent tissues than that in control group. And the patients of experimental group experienced much less postoperative bleeding than those of control group. Conclusions Both collagen matrix and free gingival graft can significantly increase the KW around implants within the 3 months post-surgery follow-up period. There were more KW gain and less shrinkage in group FGG than that in group CM. However, surgery time were reduced and the postoperative bleeding were less in group CM than in group FGG as no harvesting procedure was needed.

Dbait is a small double-stranded DNA molecule that has been utilized as a radiosensitizer to enhance the sensitivity of glioma to radiotherapy (RT). However, there is no effective drug delivery system to effectively overcome the blood-brain barrier (BBB). The aim of this study was to develop a gene delivery system by using the BBB and glioma dual-targeting and microenvironment-responsive micelles (ch-K(s-s)R8-An) to deliver Dbait into glioma for RT. Angiopep-2 can target the low-density lipoprotein receptor-related protein-1 (LRP1) that is overexpressed on brain capillary endothelial cells (BCECs) and glioma cells. In particular, due to upregulated matrix metalloproteinase 2 (MMP-2) in the tumor microenvironment, we utilized MMP-2-responsive peptides as the enzymatically degradable linkers to conjugate angiopep-2. The results showed that ch-K(s-s)R8-An micelles maintained a reasonable size (80-160 nm) with a moderate distribution and a decreased mean diameter from the cross-linking as well as exhibited low critical micelle concentration (CMC) with positive surface charge, ranging from 15 to 40 mV. The ch-K5(s-s)R8-An/pEGFP showed high gene transfection efficiency , improved uptake in glioma cells and good biocompatibility and . In addition, the combination of ch-K5(s-s)R8-An/Dbait with RT significantly inhibited the growth of U251 cells . Thus, ch-K5(s-s)R8-An/Dbait may prove to be a promising gene delivery system to target glioma and enhance the efficacy of RT on U251 cells.