Effects of Cordyceps sinensis on the Expressions of NF-κB and TGF-ß1 in Myocardium of Diabetic Rats.

Objective. To investigate the effect of Cordyceps sinensis (CS) on the expressions of NF-κB and TGF-ß1 in myocardium of streptozotocin-induced diabetic rats. Methods. A total of 53 healthy male SD rats, mice age of 8 weeks and weight of 220 ± 20 g, were randomly divided into five groups by randomized block design: normal control group (n = 10), diabetic group (n = 10), low dose of CS group (n = 12; CS 0.6 g·kg(-1)·d(-1)), middle dose of CS group (n = 11; CS 2.5 g·kg(-1)·d(-1)), and high dose of CS group (n = 10; CS 5 g·kg(-1)·d(-1)). The diabetic models with tail intravenous injection by streptozotocin (45 mg·kg(-1)). Diabetic rats were sacrificed after 8 weeks; the expressions of NF-κB and TGF-ß1 proteins and mRNA in the cardiac muscle were determined by using immunohistochemistry staining and reverse transcription polymerase chain reaction (RT-PCR) method. The data were analyzed using one factor analysis of variance. Result. The expressions of NF-κB and TGF-ß1 proteins and mRNA in the cardiac muscle of diabetic rats were significantly raised (P < 0.05), which could be decreased by CS (P < 0.05). Conclusions. The changes on the expressions of NF-κB and TGF-ß1 in myocardium may be involved in the occurrence of diabetic cardiomyopathy (DC). CS may play its role on myocardial protection by regulating the expressions of NF-κB and TGF-ß1 in myocardium.

Efficacy and safety of avandamet or uptitrated metformin treatment in patients with type 2 diabetes inadequately controlled with metformin alone: a multicenter, randomized, controlled trial.

BACKGROUND: At present, China has listed the compound tablet containing a fixed dose of rosiglitazone and metformin, Avandamet, which may improve patient compliance. The aim of this study was to evaluate the efficacy and safety of Avandamet or uptitrated metformin treatment in patients with type 2 diabetes inadequately controlled with metformin alone. METHODS: This study was a 48-week, multicenter, randomized, open-labeled, active-controlled trial. Patients with inadequate glycaemic control (glycated hemoglobin [HbA1c] 7.5-9.5%) receiving a stable dose of metformin (≥1500 mg) were recruited from 21 centers in China (from 19 November, 2009 to 15 March, 2011). The primary objective was to compare the proportion of patients who reached the target of HbA1c ≤7% between Avandamet and metformin treatment. RESULTS: At week 48, 83.33% of patients reached the target of HbA1c ≤7% in Avandamet treatment and 70.00% in uptitrated metformin treatment, with significantly difference between groups. The target of HbA1c ≤6.5% was reached in 66.03% of patients in Avandamet treatment and 46.88% in uptitrated metformin treatment. The target of fasting plasma glucose (FPG) ≤6.1 mmol/L was reached in 26.97% of patients in Avandamet treatment and 19.33% in uptitrated metformin treatment. The target of FPG ≤7.0 mmol/L was reached in 63.16% of patients in Avandamet treatment and 43.33% in uptitrated metformin treatment. Fasting insulin decreased 3.24 ± 0.98 µU/ml from baseline in Avandamet treatment and 0.72 ± 1.10 µU/ml in uptitrated metformin treatment. Overall adverse event (AE) rates and serious AE rates were similar between groups. Hypoglycaemia occurred rarely in both groups. CONCLUSIONS: Compared with uptitrated metformin, Avandamet treatment provided significant improvements in key parameters of glycemic control and was generally well tolerated. REGISTRATION NUMBER: ChiCTR-TRC-13003776.

Randomized, double blinded, placebo-controlled trial to evaluate the efficacy and safety of tolvaptan in Chinese patients with hyponatremia caused by SIADH.

To study the effect of tolvaptan on non-acute, non-hypovolemic hyponatremia in inappropriate secretion of antidiuretic hormone (SIADH) syndrome in Chinese patients. Hyponatremic SIADH patients received placebo (N = 18) or tolvaptan (N = 19) at an initial dose of 15 mg/day with further titration to 30 mg/day and 60 mg/day based on serum sodium concentrations. Randomized, double-blind, placebo-controlled trial. Primary endpoint was the change of the serum sodium from baseline to days 4 and 7. Analysis of covariance (ANCOVA) was used for statistical analysis. At day 4, average daily changes in serum sodium levels from baseline was 1.9 ± 2.9 mmol/L (1.9 ± 2.9 mEq/L) in the placebo group and 8.1 ± 3.6 mmol/L (8.1 ± 3.6 mEq/L) in the tolvaptan group; at day 7, the values were 2.5 ± 3.9 mmol/L (2.5 ± 3.9 mEq/L) and 8.6 ± 3.9 mmol/L (8.6 ± 3.9 mmEq/L) for the placebo and tolvaptan groups (ANCOVA, P < 0.001). At days 4 and 7, daily urine output and proportions of patients with normalized serum sodium were significantly superior in the tolvaptan group. The most common adverse events occurring in the tolvaptan group were dry mouth and thirst. Tolvaptan demonstrated superiority to placebo in the treatment of Chinese SIADH patients with hyponatremia by elevating serum sodium concentration with acceptable safety profile.

STAT1/3 and ERK1/2 synergistically regulate cardiac fibrosis induced by high glucose.

BACKGROUND: Hyperglycaemia promotes the proliferation of cardiac fibroblasts (CFs) and collagen synthesis in CFs. However, the molecular mechanism underlying the effects of HG on proliferation and collagen synthesis of CF, is not completely understood. OBJECTIVES: The objectives of the present study were to determine whether the STAT proteins has a functional role in high glucose-induced proliferation of CFs and collagen synthesis in vitro and whether the STAT signaling pathway and MAPK signaling pathway have synergetical effects on high glucose-mediated cardiac fibroblasts proliferation and collagen synthesis. METHODS: Rat CFs were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 or 25 mmol/L D-glucose, in the presence of absence of STAT1 inhibitor Fludarabine, STAT3 inhibitor S31-201 and ERK1/2 inhibitor PD98059. Proliferation were measured by the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, the production of Type I and III collagen was evaluated using real-time quantitative RT-PCR and ELISA, and the phosphorylation expression of STAT1 and STAT3 were analyzed by Western blot. RESULTS: High glucose treatment promoted the proliferation of cardiac fibroblasts and collagen types I and III synthesis. High glucose treatment induced STAT1 and STAT3 phosphorylation in cardiac fibroblasts, the mode and level of STAT1 and STAT3 phosphorylation were significantly different. Fludarabine and S31-201 could both inhibited high glucose stimulated proliferation of cardiac fibroblasts and collagen types I and III synthesis with different effects. Combination of Fludarabine and PD98059 or combination of S31-201 and PD98059 both exhibited stronger inhibitions on proliferation of cardiac fibroblasts and collagen types I synthesis, but the effects and functional modes are different. CONCLUSION: Both STAT1 and STAT3 mediate the proliferation of cardiac fibroblasts and collagen synthesis induced by high glucose. STAT1 and STAT3 both have synergetic effects with ERK1/2 on regulating proliferation of cardiac fibroblasts and collagen types I synthesis.

Autoimmune hypophysitis may eventually become empty sella.

Autoimmune hypophysitis (AH) is commonly believed to be a rare chronic inflammatory condition of the pituitary gland. In clinical practice, however, the disease is often seen indeed. It typically presents with hypopituitarism and pituitary mass found by MRI. We report here unusual presentations of two females with AH followed by empty sella syndrome. The two females, aged at 64 and 57-years-old, presented with anterior pituitary dysfunction, diplopia and diabetes insipidus. By MRI the two patients shared the common characteristics with diffuse homogenous contrast enhancement of the gland and increased stalk thickness. After a long period treatment with glucocorticoids, empty sella was eventually detected by MRI.

[Clinical analysis of drug-induced Pseudo-Bartter's syndrome: a report of five cases].

OBJECTIVE: To summarize the clinical characteristics and outcomes of Pseudo-Bartter's syndrome and explore its pathogenesis. METHODS: The clinical data of 5 cases of Pseudo-Bartter's syndrome at our ward from May 2008 to December 2010 was analyzed retrospectively. RESULTS: All patients were female. Long-term regimen of purgative or diuretics was prescribed. The clinical features included normotension, hypokalemic alkalosis and activation of renin-angiotensin-aldosterone. The pathological results of 3 cases of kidney biopsy showed the hyperplasia of juxtaglomerular apparatus, thickness of arteriole, infiltration of lymphocytes and monocytes and degeneration of renal tubule. Upon a definitive diagnosis, purgative or diuretics was discontinued and supplement therapy of potassium chloride initiated. The results of laboratory tests reverted to normal ranges within 4 weeks. CONCLUSION: Purgative or diuretics should be prescribed appropriately to avoid the occurrence of Pseudo-Bartter's syndrome.

[Clinical analysis of 6 cases of Bartter syndrome].

OBJECTIVE: To summarize the clinical characteristics of Bartter syndrome and investigate its pathogenesis. METHODS: The clinical data of 6 cases of Bartter syndrome at our hospital from November 2006 to May 2010 were analyzed retrospectively. RESULTS: The onset age of Bartter syndrome was 13-35 years old. The main symptoms included weakness (6/6), paralysis (1/6), numbness (5/6) and tetany (4/6). All patients had normal blood pressure. The biochemical tests showed persistent hypokalemia, metabolic alkalosis (6/6) and hyperreninemia. The pathological examination of deltoid muscle biopsy showed the swelling, degeneration and necrosis of myocytes and the deposition of immunocomplex in myolemma. And the pathological examination of renal biopsy showed the hyperplasia of juxtaglomerular apparatus (5/6) and the deposition of immunocomplex. All symptoms were relieved after a therapy of potassium supplementation or a combination of indomethacin, spironolactone and immunosuppressant. CONCLUSION: When such clinical features as weakness, paralysis, tetany, hypokalemic alkalosis and normotension are encountered, Bartter syndrome should be suspected. Serum electrolytes, blood gas analysis and activation of the renin-angiotensin-aldosterone system should be examined for a definite diagnosis. The treatment of choice includes potassium and magnesium supplementation or in combination with prostaglandin synthetase inhibitor, aldosterone antagonist and immunosuppressant. Immunologic mechanism may participate in the course of Bartter syndrome.

[The protective effects of cyclosporine A on aortic immunological injuries in STZ-induced diabetic rats].

OBJECTIVE: To investigate the autoimmune injuries of diabetic macrovascular disease (aorta) and the protective effects of immunosuppressive agent (cyclosporine A, CsA) on aortic injuries in streptozotocin (STZ)-induced diabetic rats. METHODS: STZ-induced diabetic rats were assigned randomly to 6 groups which received low (BML or AML, 1 mgxkg(-1)xd(-1)), middle (BMM or AMM, 4 mgxkg(-1)xd(-1)) or high (BMH or AMH, 8 mgxkg(-1)xd(-1)) dose of CsA from 1 week before or after STZ for 8 weeks. Diabetic rats without any treatment, insulin-treated diabetic rats and normal rats were also monitored simultaneously and served as control groups. The pathologic abnormalities of the aorta were verified by HE, Masson staining and electronmicroscopy. The depositions of immunoglobulins (IgG, IgM and IgA) were determined by immunohistochemistry and immunofluorescence methods. RESULTS: At the end of study, lymphocytes infiltration and collagen content (26 582 +/- 6901) were significantly higher in diabetic aorta than those in non-diabetic aorta (Collagen: 7482 +/- 3491, P < 0.01). The deposited IgG and IgA were also significantly increased in diabetic aorta compared with non-diabetic aorta (IgG: 11 789 +/- 2491 vs. 2518 +/- 1066, P < 0.01; IgA: 17 430 +/- 3159 vs. 1135 +/- 758, P < 0.01). These changes were not affected by insulin while CsA intervention significantly reduced aortic collagen content (BMH: 13 518 +/- 5440, P < 0.01 vs. STZ) and immunoglobulin deposition (BMH: IgG: 7584 +/- 4462; IgA: 6176 +/- 1900, all P < 0.01 vs. STZ). These immunoglobulin deposition changes were confirmed by results of immunofluorescence. Aortic collagen accumulation was positively correlated to aortic immunoglobulin deposition (IgG, r = 0.556, P < 0.01; IgA, r = 0.661, P < 0.01). CONCLUSIONS: Our data suggest that the autoimmune injuries might be a promoting factor in the pathogenesis of the diabetic macrovascular disease which could lead to the development of macrovascular disease. Immunosuppressive agent, such as CsA, could inhibit the abnormal deposition of immunoglobulins and therefore, delay the development of diabetic macrovascular disease in this model.

[Regulation of receptor activator of nuclear factor kappaB ligand and osteoprotegerin expression by prostaglandin E2 in osteoblastic-like cells].

OBJECTIVE: To investigate the regulation of receptor activator of nuclear factor kappaB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression by prostaglandin E2 (PGE2) in osteoblastic-like cells and involved signaling pathways. METHODS: Rat UMR106 osteoblast-like cells were cultured and treated with various dose of PGE2 or regulators of different signaling pathways such as protein kinase A, protein kinase C, ERK-MAPK and calcium/calmodulin pathways for different period of time,the cells were then harvested at indicated time points, total RNA were isolated and RANKL/OPG mRNA expression were studied by real-time PCR. RESULTS: PGE2, Forskolin and db-cAMP increased RANKL mRNA by 2.8 times (P = 0.002), 2.2 times (P = 0.006) and 2.1 times (P = 0.005) respectively, and inhibited OPG mRNA expression by 12% (P < 0.01), 85% (P = 0.005) and 70% (P = 0.013) respectively, while RANKL and OPG mRNA expression were down-regulated by A23187 by 58% (P = 0.002) and 53% (P = 0.017) respectively. As for inhibitory experiments, the stimulatory effects of PGE2 on RANKL mRNA expression could be inhibited only by KT-5720 by 53% (P < 0.01), while the other inhibitors did not have any effect at all. The inhibitory effects of PGE2 on OPG mRNA expression were partially blocked by KT-5720 by 47% (P = 0.01), verapamil by 38% (P = 0.029) and W7 by 43% (P < 0.01) respectively, while KN-62, chelerythrine and PD98059 had no effects. CONCLUSION: PGE2 can up-regulate the expression of RANKL but down-regulate the expression of OPG. The up-regulation of RANKL mRNA expression by PGE2 was mediated through the activation of PKA signaling pathway while PGE2 induced down-regulation of OPG mRNA expression was predominantly mediated via PKA as well as the Ca2+ /Calmodulin signaling pathways.

[The effects of cyclosporine A on immunoglobulins deposition in retina of streptozotocin-induced diabetic rats].

OBJECTIVE: To study the autoimmune injuries on diabetic retinopathy (DR) and the protective effects of immunosuppressive therapy with cyclosporine A (CsA) on the DR of streptozotocin (STZ)-induced diabetic rats. METHODS: STZ-induced diabetic rats were randomly divided into 3 groups: group 1: diabetic group without any treatment; group 2: insulin-treated group; group 3: CsA-treated group which was further divided into 2 subgroups: subgroup A: CsA was given 1 week before hyperglycemia appeared and subgroup B: CsA was given one week after hyperglycemia appeared. Subgroup A and subgroup B were further subdivided into 3 groups respectively, based on the dose of CsA (1, 4 and 8 mg x kg(-1) x d(-1)). As a control group, normal rats were also simultaneously monitored. The pathologic changes in the retina were investigated with HE stain and the deposition of immunoglobulins was detected with immunohistochemistry and immunofluorescent microscopy. RESULTS: After 8 week, the deposition of IgG, IgA and IgM was quite significant in the retina of diabetic rats. The data also suggested that insulin treatment had no effects on the DR. In contrast,with CsA intervention, the deposition of immunoglobulins on the retina of diabetic rats vanished. CONCLUSIONS: Autoimmune injuries were shown, in the present study, to play a critical role in the pathogenesis of diabetic retinopathy. Immunosuppressive treatment with CsA showed protective effects by inhibiting the deposition of immunoglobulins in the retina of diabetic rats.

Open-label, randomized, multiple-center, parallel study comparing glycemic responses and safety profiles of Glucerna versus Fresubin in subjects of type 2 diabetes mellitus.

Since the increase of prevalence of type 2 diabetes mellitus (T-2DM), the replacing quickly absorbed carbohydrates with a fat source rich in monounsaturated fatty acid to provide improved glycemic control in these patients has become an important assistant therapy. In the present study, we compared glycemic response and safety of two nutritional products, Glucerna and Fresubin, in Chinese subjects with T-2DM. Overall, 203 T-2DM subjects were randomly assigned (1:1) to either Glucerna or Fresubin. The primary endpoint was the adjusted area under the curve (adj-AUC) for plasma glucose at 0-240 min. Blood samples were collected at 0, 30, 60, 90, 120, 180, and 240 min to compare the adjusted area under the curve (AUC) for the change in plasma glucose or insulin from 0 to 240 min. Adjusted peak values and times of glucose and insulin responses and adjusted glucose and insulin values were collected at the same time points. Safety parameters were also evaluated. The adjusted AUC for the change in plasma glucose in the Glucerna group was significantly lower than in Fresubin group (5.60 +/- 5.88 mmol/l*h vs. 7.97 +/- 6.32 mmol/l*h, P = 0.0061), as was the adjusted peak value of glucose (3.51 +/- 2.04 mmol/l vs. 4.69 +/- 1.99 mmol/l, P < 0.0001). Glucerna subjects had a longer adjusted peak time to insulin response compared to Fresubin subjects (105.00 +/- 43.4 min vs. 88.81 +/- 37.69 min, P = 0.0050). Glucerna subjects also experienced more gradual changes in glucose and insulin values. In conclusion, Glucerna provided better control of postprandial plasma glucose and insulin levels in Chinese subjects with T-2DM. Variation of postprandial glucose tended to be relatively stable after patients took Glucerna. Study results suggest that Glucerna may be beneficial in the reduction of postprandial glycemia.

[Relationship between gastrointestinal dyskinesis and gastrointestinal neurons in diabetic mellitus: experiment with rats].

OBJECTIVE: To investigate the relationship between gastrointestinal dyskinesis and histologic changes of gastrointestinal myenteric plexus cholinergic and nitrergic neurons in STZ-induced diabetic rats. METHODS: 45 SD rats were randomly divided into control group, diabetic group and insulin group. 16 weeks after diabetic model established, gastrointestinal motility of rats was measured and histologic changes of myenteric plexus cholinergic neuron and nitrergic neuron was observed. RESULTS: Compared with control group, gastrointestinal motility of diabetic group was markedly slow (P < 0.01), the myenteric plexus cholinergic neuron counting of gastric antrum and small intestine were significantly decreased (6.6 +/- 2.9 vs 15.7 +/- 3.8 15.6 +/- 10.3 vs 22.6 +/- 7.4, P < 0.01), yet the number of nitrergic neuron only markedly reduced in gastric antrum (5.3 +/- 1.2 vs 11.8 +/- 2.2, P < 0.01). The gastrointestinal mobility, gastric antrum nitrergic neuron and small intestine cholinergic neuron counting of insulin group were markedly higher than that of diabetic group (P < 0.05), yet lower than that of control group (P < 0.01). CONCLUSION: The gastrointestinal dyskinesis of STZ-induced diabetic rats might be associated with lesions of gastrointestinal myenteric plexus cholinergic neuron and nitrergic neuron. Insulin intensive therapy can partly ameliorate diabetic gastrointesternal dyskinesis.

[Effects of OGP(10-14) and its derivative G38I on proliferation and differentiation of rat osteoblasts in vitro].

OBJECTIVE: To investigate the effects of the carboxyl end of osteogenic growth peptide (OGP)-OGP((10-14)) and its derivative G38I on the proliferation and differentiation of osteoblasts (OBs). METHODS: Osteoblasts were isolated from the calvariae of newborn SD rats and cultured to G3. OGP((10-14)) or G38I of the concentrations of 10(-15) to 10(-7) mol/L were added to culture medium for 48 hours respectively. The number of cells was counted and MTT analysis was used to examine the proliferation of the cells. The ultrastructure of cells was investigated by electron microscopy. The osteoblasts of G3 were divided into experimental groups, treated with OGP((10-14)) or G38I of the concentration of 10(-11) mol/L for 48 hours, and control group. The alkaline phosphatase activity in the culture medium was measured. The protein expression level of type-I collagen was evaluated by immunohistochemistry. The core binding factor 1 (Cbfa1) and type-I collagen mRNA level of osteoblasts were determined by RT-PCR. RESULTS: With a biphasic effect on, OGP((10-14)) and G38I stimulated the number enhancement of OBs dose-dependently at low concentration and inhibited it at high concentration. The numbers of OB were the highest (37 x 10(4)/ml +/- 7 x 10(4)/ml and 30 x 10(4)/ml +/- 5 x 10(4)/ml respectively) when treated by OGP((10-14)) or G38I of the concentration of 10(-11)mol/L. The rough endoplasm net was flourishing and the secreting vesicle was abounding in the experimental cells. There was calcium crystal in the control cells. The activity of alkaline phosphatase in the culture medium of the OGP (10(-14)) and G38I groups were higher than that in the control group (4.47 U/g and 3.82 U/g vs 2.21 U/g). The protein expression level of type-I collagen was higher and the mRNA levels of Cbfa1 and type-I collagen were higher in the OGP((10-14)) and G38I groups were increased in the experimental groups in comparison with the control group (P < 0.05, P < 0.01, and P < 0.05). CONCLUSION: They stimulated cell number enhancement dose dependently at low concentration and followed by inhibition at high concentration. Just as the native OGP, OGP((10-14)) and its derivative G38I stimulate the proliferation of osteoblasts, and improve their activity, up-regulate the Cbfa1 and type-I collagen mRNA expression levels and increase the collagen synthesis, thus promoting the differentiation and osteogenic effect of osteoblasts.

[Impacts of osteoclast-like cells cultured on bone wafer and their sub-cellular structures on osteoblast].

OBJECTIVE: To investigate the effects of osteoclast-like cells (OLC) and its sub-cellular structures on the osteoblast (OB) differentiation and function. METHODS: Spleen cells from C57 mice administrated with 5-fluorouracil were induced by IL-3, 6 and granulocyte-macrophage colony stimulating factor (GM-CSF), and 1alpha, 25-(OH)(2)D(3) to obtain massive OLCs. These OLC cells were cultured in culture fluid and on bone wafers (called bolcs). Osteoblasts were cultured and added with NaF, OLCs of two kinds, culture fluid free of OLC, and sub-unit structures such as nucleus, mitochondria, and cytoplasma from OLCs for 5 days. The proliferation rate of OBs was measured by MTT method and the alkaline phophatase (ALP) activity was measured by PNPP method. Immunochemistry was used to detect the core-binding factor alpha1 (Cbfalpha1) in the OBs, Enzyme linked immunosorbent assay was used to measure the osteocalcin. RESULTS: The OB number was lower in the OLC (1.288 +/- 0.039), OLC cytoplasm (1.138 +/- 0.024), 50% OLC culture fluid (1.203 +/- 0.033), 50% OLC culture medium of OLCs cultured on bone wafer (1.128 +/- 0.028) in comparison with the pure OB group (1.393 +/- 0.016, all P < 0.05). The increase functions of OBs by OLC cultured on bone wafer and their nucleus and mitochondria were all more significant than those of the OLCs not cultured on bone wafer. The ALP activity was increased in the NaF (1.027 +/- 0.024), OCL cytoplasm (1.850 +/- 0.033), 50% OLC medium (2.074 +/- 0.065), 50% OLC medium of OLCs cultured on bone wafer (1.718 +/- 0.048), and mitochondria and cytoplasm of the OLC cultured on bone wafer groups (1.246 +/- 0.037, all P < 0.05). NaF (0.0825 +/- 0.0025), OLCs (0.0775 +/- 0.0025), nucleus (0.0775 +/- 0.0025), mitochondria (0.0875 +/- 0.0025), and cytoplasm of OLCs (0.1100 +/- 0.0007), 50% OLC medium (0.0900 +/- 0.0000), 50% OLC medium of OLCs cultured on bone wafer (0.1200 +/- 0.0041), OLCs cultured on bone wafer and nucleus, mitochondria, and cytoplasm of OLCs cultured on bone wafer all significantly increase the oeteocalcin activity of OBs (0.525 +/- 0.0063, all P < 0.05). NaF (57.6% +/- 2.6%), OLC cytoplasm (45.3% +/- 4.7%), 50% OLC medium (46.6% +/- 3.3%), 50% medium of OLCs cultured on bone wafer (54.0% +/- 2.1%), OLCs cultured on bone wafer (44.8% +/- 3.0%), and cytoplasm of OLCs cultured on bone wafer (48.7% +/- 3.5%) all significantly increased the Cbfalpha1 protein in the OBs (32.8% +/- 4.5%, all P < 0.05). CONCLUSION: The sub-cellular elements of OLC and the supernatant of OLC culture media free of OLC promote the functions of OB, especially the OLCs cultured on bone wafer.