ObjectiveTo observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.MethodsThirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.ConclusionThe effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis

Objective To observe the expression of matrix gla protein(MGP) mRNA in primary osteoblasts of Sprague-Dawley (SD) rat in vitro after treatment with anti-osteoporosis agents [vitamin K2,PTH,1,25 (OH)2D3,and alendronate],and to investigate the potential role of MGP in the pathogenesis of osteoporosis.Methods Primary osteoblasts(OBs) were derived from sequential trypsin/collagenase-digested calvaria isolated from newborn SD rat (postnastal day 1-3).OBs of the second generation were identified by Van Gieson collagen staining,alkaline phosphatase(ALP) staining and calcified nodules staining.OBs of the fourth generation were selected to interfere with vitamin K2,PTH,1,25 (OH)2D3,and alendronate,then cultured for 24 h in mediums which contained various concentrations of vitamin K2 (10-7,10-6,and 10-5 mol/L),PTH (10-9,10-8,and 10-7 mol/L),1,25 (OH) 2D3(10-10,10-9,and 10-8mol/L),alendronate(10-6,10-5,and 10-4mol/L).After being cultured for 24 h,total RNA was extracted and examined by real-time quantitative RT-PCR.Results The primary cultured cells had typical morphological characters of osteoblast.van Gieson collagen staining,ALP staining,and calcified nodules staining were all positive.Vitamin K2,PTH,1,25 (OH)2D3,and alendronate could modulate the expression of MGP mRNA in osteoblasts in a dose-dependent fashion.MGP mRNA expressions were 2.56-fold,2.12-fold,and 1.57-fold with 10-5,10-6,and 10-7 mol/L of vitamin K2 treatment,respectively.The expressions were 6.78-fold,5.31-fold,and 2.23-fold with 10-7,10-8,and 10-9mol/L of PTH(1-34) treatment,8.93-fold,6.95-fold,and 3.47-fold with 10-8 10-9,and 10-10mol/L of 1,25 (OH)2D3 treatment,and 3.47-fold,2.49-fold,and 1.98-fold with 10-4,10-5,and 10-6mol/L of alendronate treatment.Conclusion Vitamin K2,PTH,1,25 (OH)2D3,and alendronate all canregulate MGP mRNA expression in calvarial osteoblasts in a dose-dependent manner.MGP seems to be a potent target of anti-osteoporosis agents,and involved in the pathogenesis of osteoporosis.

Objective: To investigate the effect of allogeneic red blood cell transfusion on T lymphocyte subsets and natural killer (NK) cells in peripheral blood of patients with gastric cancer during perioperative period. Methods: 50 patients with gastric cancer in our hospital from January 2017 to December 2017 were randomly divided into control group (n=25) and observation group (n=25). Perioperative allogeneic erythrocyte transfusion was performed in the observation group, while no allogeneic erythrocyte transfusion was performed in the control group. Blood samples were taken from two groups before and after operation 3 d, and serum T lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺, CD4⁺ /CD8⁺ ) and NK cell levels were measured. T cell subsets and NK cell levels of patients in observation group, control group and observation group at different clinical stages before and after operation were counted. According to blood transfusion volume, the patients in the observation group were divided into <4 U group and <4 U group. T cell subsets and NK cell levels before and after operation were compared between the two groups. Results: There was no significant difference in CD3⁺, CD4⁺, CD8⁺, CD4⁺ /CD8⁺, NK cells between the two groups before operation (P>0.05). The levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells in the two groups after operation were lower than those before operation and CD8⁺ levels were higher than those before operation. The levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells in the observation group were lower than those in the control group, and the levels of CD8⁺ in the observation group were higher than those in the control group (P<0.05). The levels of CD3⁺, CD4⁺, CD8⁺, CD4⁺ /CD8⁺, NK cells in patients with stage Ⅰ-Ⅱ and stage Ⅲ-Ⅳ had no significant difference before operation (P>0.05). After operation, the levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells in the two groups were lower and CD8⁺ levels were higher than those before operation. The levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells in stage Ⅰ-Ⅱ patients were higher than those in stage Ⅲ-Ⅳ patients, and CD8⁺ levels were lower than those in stage Ⅲ-Ⅳ patients (P< 0.05); There was no significant difference in the levels of CD3⁺, CD4⁺, CD8⁺, CD4⁺ /CD8⁺, NK cells between the <4 U group and <4 U group before operation (P>0.05). After operation, the levels of CD8⁺ in the two groups were higher than those before operation, while the levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells were lower than those before operation; The levels of CD3⁺, CD4⁺, CD4⁺ /CD8⁺, NK cells in the <4 U group were higher than those in >4 U group, and CD8⁺ levels were lower than those in >4 U group (P<0.05). Conclusions Perioperative allogeneic red blood cell transfusion can affect T cell subsets and NK cell content in peripheral blood of patients with gastric cancer, and there are some differences in the effects of different clinical stages of gastric cancer patients, which can cause the suppression of cellular immune function.