Objective To observe the effect of fluvastatin on urinary albumin excretion rate(UAER)of microalbuminuria in type 2 diabetes patients.Methods Type 2 diabetes patients with mieroalbuminuria were randomly divided into fluvastatin goup(n=126)and control group(n=129).Fluvastatin group was given fluvastatin 40 mg each night.Follow up was conducted for 1.5 years.The UAER and glomerular filtration rate(GFR)as well as blood lipid level before and after therapy were compared.Results UAER in fluvastatin group was significantly lower than that in control group[(59.6±10.5)vs(87.5±12.3)mg/min,P＜0.05]before therapy[(104.4±25.2)vs(110.6±19.7)mg/min,P＜0.05],which was independent of its lowering-lipid effect.But GFR had no signifimicroalbuminuria which is independent of lowering-lipid effect in type 2 diabetic nephropathy patients and delay the progress of diabetic nephropathy.

Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.

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 clinical outcomes of pedicle screw fixation assisted with the 3D printing positioning guide template for treatment of unstable atlas fractures.Methods A retrospective case series review was made on 10 patients with unstable atlas fractures undergone direct posterior C1 pedicle screw fixation assisted with the 3D printing drill guide template from September 2012 to May 2015.There were 7 males and 3 females,with a mean age of 52.6 years (range,23-75 years).All patients complained of neck pain,stiffness and decreased range of motion without neurologic deficit.Preoperative skull traction was used routinely.After the three-dimension reconstruction of cervical vertebrae,ideal trajectory for C1 pedicle screws was designed with a complementary basal template for posterior surface of atlas corresponding anatomical structure.Then the drill guide template was materialized in a rapid prototyping machine and used during operation.Start point and direction of the ideal and actual trajectories were measured after matching the position of the pre-and post-operative patients' cervical spine.Safety of pedicle screw fixation was assessed in the transverse and sagittal planes of CT scan.Operation time and blood loss were recorded.Visual Analogue Scale (VAS) of neck pain was recorded before operation and 3 months after operation.Clinical efficacy,fracture reduction,stability and surgical complications were reviewed at the follow-up.Results A total of 20 screws were inserted safely.No significant differences existed in deviation of entry point and direction between ideal and actual trajectories (P ＞0.05).Operation time was 60-90 min (mean,75 min) and intraoperative blood loss was 110-300 ml (mean,160 ml).No spinal cord or vertebral artery injury was noted during operation.All patients were followed up for 12-36 months (mean,20.5 months).VAS was improved from preoperative 7.3 (6.3-9.5) points to 1.4 (0.3-2.5) points 3 months after operation (P ＜ 0.05).All patients had normal range of motion of the cervical spine 3 months after operation.Bony fusion was achieved 6 months after operation.At the follow-up,good cervical alignment was maintained with no instrument failure and C1.2 instability.Conclusion For treatment of unstable atlas fractures,direct posterior C1 pedicle screw fixation assisted with the 3D printing drill guide template can improve the precision of screw placement,reduce complications,and preserve the function of the occipital-atlantoaxial junction.

Objective To investigate the effect of glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide on hypoxia/reoxygenation (H/R)-induced cardiomyocytes death under high glucose condition and the potential mechanisms.Methods H9C2 cardiomyocytes were divided into 4 groups:normal glucose (N,5 mmol/L),high glucose (G,20 mmol/L),high glucose in combination with liraglutide (L,100 nmol/L),high glucose in combination with liraglutide and wortmannin (W,25 nmol/L).The apoptosis of H9C2 was detected by TUNEL assay.Nitric oxide synthetase(eNOS),nitric oxide (NO) and reactive oxygen(ROS) in supernatants were measured by enzymatic analysis,p-PI3K,PI3K,p-Akt,Akt,Bcl-2,caspase-3 were examined by western blotting.Results Compared with cells in N group,the apoptosis of H9C2 cells induced by H/R was markedly increased [(15.79 ± 3.92) ％ vs (9.74 ± 1.14) ％,P =0.028] in G group.The same was true for ROS [(489.63 ±21.01) U/ml vs (338.50 ±43.60) U/ml,P ＜0.001] and caspase-3 levels (1.87 ±0.03 vs 1.15 ±0.04,P ＜0.001),but not for Bcl-2 protein expression (1.79 ± 0.06 vs 1.89 ±0.03,P =0.047).Pretreatment of cells with liraglutide (100 nmol/L) prevented the cell death induced by high glucose and H/R together with decrease of ROS and caspase-3 levels and increase of Bcl-1 expression.Moreover,treatment of cells with liraglutide also significantly increased phosphorylation of PI3K and Akt (p-PI3K/PI3K:0.87 ± 0.07 vs 0.59 ± 0.09,P =0.002;p-Akt/Akt:0.34 ± 0.01 vs 0.08 ± 0.01,P＜0.001),eNOS[(41.29 ±0.56) μmpl/L vs (37.20 ±0.52)μxmpl/L,P ＜0.001]and NO [(31.24 ±0.40) μmpl/L vs (26.66 ±0.53) μmpl/L,P ＜0.001] levels.Furthermore,addition of PI3K/Akt inhibitor wortmanin markedly inhibited the expression of p-PI3K/PI3K,p-Akt/Akt,reversed the changes of eNOS,NO,caspase-3 and Bcl-2 by liraglutide,and abolished the protective effect of liraglutide on cell apoptosis.Conclusions GLP-1 receptor agonist liraglutide treatment could alleviate cardiomyoeytes apoptosis induced by high glucose and H/R through the activation of PI3K-Akt-eNOS-NO signaling pathway and inhibition of oxidative stress.

Objective To compare the clinical efficacy and fusion rate of unilateral and bilateral C1 and C2 pedicle screw fixation of unstable Jefferson fractures.Methods This retrospective casecontrol study enrolled 22 patients with unstable Jefferson fractures admitted between April 2012 and May 2015.There were 18 males and four females,with the mean age of 52.9 years (range,35-67 years).Mean preoperative visual analogue scale (VAS) was 6.09 points (range,4-8 points).According to the American spinal injury association (ASIA) classification,two patients were rated grade D and one patient grade C.Mean Japanese orthopedic association (JOA) score was 12.3 points.Bilateral C1 and C2 pedicle screw fixation was performed for 15 patients (bilateral group).Advantage side unilateral C1 and C2 pedicle screw fixation was performed for seven patients with extremely unstable fracture or narrow pedicle (unilateral group).Operation time,blood loss and surgical complications were recorded.VAS was used to evaluate the improvement of neck pain after operation.ASIA classification and JOA score were used to assess nerve function recovery.Atlanto-dental interval (ADI),srew position and bone fusion were evaluated after operation.Results All patients successfully completed the operation.Operation time was (119.5 ±21.2)min,and blood loss was (280.1 ±83.1)ml.A total of 74 screws were placed and CT scan showed satisfactory position of the screws.No complications were noted either during the operation or after surgery.All patients were followed up for mean 20.7 months (range,13-33 months).VAS was improved in both groups after operation (P ＜ 0.01),and there was no significant difference between the two groups (P ＞ 0.05).Two patients with ASIA grade D in bilateral group were improved to ASIA grade E after operation.One patient with ASIA grade C in unilateral group was improved to ASIA grade D after operation.JOA score increased to mean 15.7 points at last follow-up.ADI were decreased in both groups after operation(P ＜0.05),but there was no significant difference between the two groups (P ＞ 0.05).All patients had bony fusion 6 months after operation,with similar fusion rate between the two groups (P ＞ 0.05).Conclusion Advantage side unilateral screw fixation can be used for the patients with bilateral C1 and C2 pedicle screw fixation failure,for the technique can improve cervical pain and provide relatively high stability and fusion rate.

Objective To compare the safety and efficacy of insulin degludec (IDeg) with those of insulin glargine (IGlar) in insulin-naive subjects with type 2 diabetes (T2DM).Methods This was a 26-week,randomized,open-label,parallel-group,treat-to-target trial in 560 Chinese subjects with T2DM (men/women:274/263,mean age 56 years,mean diabetes duration 7 years) inadequately controlled on oral antidiabetic drugs (OADs).Subjects were randomized 2:1 to once-daily IDeg (373 subjects) or IGlar(187 subjects),both in combination with metformin.The primary endpoint was changes from baseline in glycosylated hemoglobin(HbA1c) after 26 weeks.Results Mean HbA1c decreased from 8.2％ in both groups to 6.9％ in IDeg and 7.0％ in IGlar,respectively.Estimated treatment difference (ETD) of IDegIGlar in change from baseline was-0.10％ points (95％ CI-0.25-0.05).The proportion of subjects achieving HbA1c ＜ 7.0％ was 56.3％ and 49.7％ with IDeg and IGlar,respectively [estimated odds ratio of IDeg/IGlar:1.26 (95 ％ CI 0.88-1.82)].Numerically lower rateof overall confirmed hypoglycaemia and statistically significantly lower nocturnal confirmed hypoglycemia were associated with IDeg compared with IGlar,respectively [estimated rateratio of IDeg/IGlar 0.69 (95％ CI 0.46-1.03),and 0.43 (95％ CI 0.19-0.97)].No differences in other safety parameters were found between the two groups.Conclusions IDeg was non-inferior to IGlar in terms of glycaemic control,and was associated with a statistically significantly lower rate of nocturnal confirmed hypoglycaemia.IDeg is considered to be suitable for initiating insulin therapy in Chinese T2DM patients on OADs requiring intensified treatment.Clinical trail registration Clinicaltrials.gov,NCT01849289.

Objective To compare the clinical effects of posterior pedicle screw fixation combined with anterior monosegmental or bisegmental fusion reconstruction in patients with unstable Denis type B thoracolumbar burst fractures.Methods A retrospective case-control analysis was made on 62 cases of Denis type B thoracolumbar burst fractures treated from June 2010 to June 2014.There were 51 males and 11 females,aged 19-55 years (mean,35.1 years).The injury causes included fall from height in 32 cases,traffic accidents in 24,hit by heavy objects in 6.Fifty cases were with monosegmental burst fractures,and 12 with monosegmental burst fractures combined with other vertebral compressive fractures.The burst fracture levels were at T~in 8 cases,T12in 20,L1 in 24,L2 in 6,and L3 in 4.According to the different surgical methods,the patients were divided into two groups:Group A (n =30,treated with posterior pedicle screw fixation combined with anterior monosegmental fusion reconstruction) and Group B (n =30,treated with posterior pedicle screw fixation combined with anterior bisegmental fusion construction).The operation time and intraoperative blood loss were compared between the two groups.Visual analogue scale (VAS),Oswestry disability index (ODI),and Frankel scale of neurologic function were compared at last follow-up.The changes of anterior height of fracture vertebrae and Cobb angle of the two groups were also compared preoperatively,postoperatively and at last follow-up.Implant looseness and breakage,titanium mesh tilt,and bone fusion were recorded postoperatively.Results All patients were followed up for 18-24 months (mean,20.8 months).The operation time and blood loss in Group A was (208.2 ± 15.6) min and (598.3 ± 55.3) ml,respectively.The operative time and blood loss in Group B was (260.1 ± 17.4)min and (662.2± 58.3)ml,respectively.There were significantly statistical differences between two groups in operation time and blood loss (P ＜0.05).There were no statistical differences between two groups in terms of Cobb angle restoration and fracture height restoration rate after surgery,Cobb angle loss and fracture vertebral body front height loss rate at final follow-up (P ＞ 0.05).At the final follow-up,VAS in Group A was (2.5 ± 0.8) points,less than (3.2 ± 1.1) points in Group B (P ＜ 0.05);ODI in Group A was (20.3 ± 5.8) points,less than (28.2 ± 5.1) points in Group B (P ＜ 0.05).Frankel scale was improved to some degree after operation (P ＜ 0.05).One case did not achieve bone fusion in Group A versus three cases in Group B (P＜0.05).No implant Loosening,titanium mesh tilt or settlement was found in Group A,while there were eight cases of titanium mesh tilt or settlement in Group B (P ＜0.05).Conclusions For unstable Denis type B thoracolumbar burst fractures,posterior pedicular fixation combined with anterior monosegmental fusion or bisegmental fusion can achieve equal clinical effects.However,posterior pedicular fixation combined with anterior monosegmental fusion can have less injury,higher bone fusion rate,better function restoration,and less incidence of complication.

Objective:To evaluate the feasibility of C 2 laminar screw fixation with double holes, and to explore a new method to judge the safe position of screws during operation. Methods:There were 22 axial specimens, 11 males and 11 females, aged from 26-69 years (mean 47.2 years). The Dicom format of Cervical spine specimens were obtained by CT scanning. It was transferred into the Mimics software for reconstructing the three-dimensional cervical spine model. The central horizontal plane of the axial lamina was cut off as the measuring plane by using the segmentation function. The cortical bone of the start and end lamina was removed naming hole 1 and 2. The distances of D1 and D2 are measured at the trajectory positions of maximum safe ventral inclination, parallel, or maximum safe dorsal inclination, with each entry point (A, M, P). A paired comparison was performed for each group.Results:Forty-four groups of parallel, introversion and extroversion screw path parameters were obtained from 22 axial specimens, and there was no significant difference between the left D1 and D2 groups in each group A, M, P [group A: 3.72±0.95 mm, 3.37±1.24 mm; group M: 2.29±0.72 mm, 1.94±0.58 mm; group P: 1.17±0.44 mm, 0.86±0.69 mm] ( t=1.051, 1.776, 1.777; P>0.05), while the right D1 and D2 had statistical significance[group A: 4.44±1.20 mm, 3.36±1.37 mm; group M: 3.01±0.76 mm, 1.97±0.90 mm; group P: 2.06±0.73 mm, 1.00±0.87 mm]( t=2.781, 4.141, 4.378; P<0.05). In introversion, there was no significant difference in left D1 and D2 in group M [3.11±0.92 mm, 3.79±1.36 mm] ( t=1.942, P=0.058), and was no significant difference in right D1 and D2 in group M [3.79±1.20 mm, 3.69±1.55 mm] ( t=0.239, P=0.812). In group P, the left D1 and D2 were compared, and the results had statistically differences [2.67±0.77 mm, 4.25±1.39 mm] ( t=4.644, P=0.000). In group P, the right D1 and D2 were compared, and the results were statistically different [2.57±1.14 mm, 3.94±1.53 mm] ( t=3.368, P=0.001). In extroversion, the left D1 and D2 in group A and M were compared, and the results had statistically differences [group A: 3.44±1.05 mm, 1.22±0.71 mm; group M: 2.26±0.73 mm, 0.90±0.75 mm] ( t=8.215, 6.095; P<0.05); the results of group A and group M were statistically different between groups of D1 and D2 on the right side[group A: 4.56±1.51 mm, 1.48±0.97 mm; group M: 2.96±1.12 mm, 1.06±0.75 mm] ( t=8.049, 6.611; P<0.05). Conclusion:The double hole screw technique of the C 2 lamina can judge the safe position of the screw in the operation without additional fluoroscopy.