Objective To investigate the value of brain ~ 99mTc-TRODAT-1 SPECT in differentiation of essential tremor (ET) from Parkinson's disease (PD) at early stage. Methods SPECT imaging with ~ 99mTc-TRODAT-1 was conducted in 28 patients with PD at early stage, 15 patients with ET, 8 patients with ET combined with PD and 15 healthy subjects. The interesting regions including bilateral striatum (ST) and occipital lobe (OC) were figured, and the radioactivity counts of these three regions were obtained by computer automatically. The ratio of specific radioactivity uptake and asymmetry indexes of the four groups were calculated and compared.Results The ratio of specific radioactivity uptake was 0.58?0.16 (left side) and 0.56?0.32 (right side) in healthy group, 0.55?0.22 (left side) and 0.56?0.24 (right side) in ET group. There was no significant difference between the two groups. The ratio of specific radioactivity uptake was 0.44?0.33 (left side) and 0.45?0.18 (right side) in ET-PD group, 0.40?0.33 (side contralateral to onset or more severe limbs) and 0.51?0.12 (ipsilateral side) in PD group. The ratios of specific radioactivity uptake in PD and ET-PD groups were significantly lower than those in ET and healthy groups ( P

Objective To investigate the inhibitory effect of the lentiviral vector (LV)-mediated RNA interference (RNAi) targeting HIF-1α on the expression of HIF-1α and Glut-1 in human pancreatic cancer Patu8988 cells.Methods The RNAi targeting HIF-1α was combined to LV,and transfected into Patu8988 cells.The Patu8988 cells transfected with the empty vector and exposed to 0.5％ O2 for 4 h served as hypoxia negative control,the Patu8988 cells not transfected with vector and exposed to 0.5％ O2 for 4 h as hypoxia blank control,and the Patu8988 cells transfected with LV-RNAi-HIF-1α and exposed to 0.5％O2 for 4 h as experimental group.The expression of HIF-1α was measured by RT-PCR and Western blot respectively.The expression of Glut-1 was measured by RT-PCR.Each group was compared according to oneway analysis of variance and two-sample t test.Results After transfection with LV-RNAi-HIF-1α,HIF-1α mRNA expression decreased by 65.1％ (0.209/0.321) and 80.6％ (0.791/0.982) (t=10.52,15.24,both P＜0.05) under normoxia and hypoxia conditions,meanwhile with the empty vector,HIF-1α mRNA expression decreased by 0.6％ (0.002/0.321) and 7.2％ (0.071/0.982) (t =5.26,7.38,both P＜0.05).Under hypoxia conditions,the protein of HIF-1α in experimental group cells (0.159±0.010) was down-regulated obviously compared to the negative control group (0.745± 0.012) and the blank control group (0.711 ± 0.023)(F=35.52,t =6.72,10.56,all P＜0.05).The expression of Glut-1 mRNA in experimental group cells (0.040±0.003) decreased obviously compared to the negative control group (0.054±0.003) and blank control group (0.062±0.004) (F=35.28,t=5.94,8.55,all P＜0.01).Conclusion Gene silencing of HIF-1α using LV-mediated RNAi can inhibit the expression of HIF-1α and decrease the expression of Glut-1mRNA in Patu8988 cells.

Objective To investigate the relationship of the changes of bone density and biochemical marker of bone turnover with osteoporosis in the elderly with type 2 diabetes mellitus(T2DM). Methods A total of 102 elderly patients with type 2 diabetes and 42 healthy subjects (normal controls) were included in this study.The bone mineral density (BMD) of lumbar vertebrae and femur were measured by dual-energy X-ray absorptiometry (DXA).The T2 DM subjects were divided into two groups:osteoporosis group (56 cases) and non-osteoporosis group (46 cases).The levels of total procollagen type Ⅰ amino-terminal propeptide (TP Ⅰ NP),β-isomerized carboxyterminal propeptide (β-CTx),parathyroid hormone (PTH),serum calcium (Ca2+),serum phosphorus (P),alkaline phosphatase (ALP),fasting blood glucose(FBG),glycosylated hemoglobin (HbAlc) and body mass index (BMI) were detected. The correlations between BMD of the osteoporosis group and other related factors were analyzed. Results The BMD values of lumbar vertebrae and femur neck,troch,ward's triangle in the osteoporosis group were decreased as compared with control(t=9.006,6.347,7.387,5.321,P＜ 0.01) and the serum TP Ⅰ NP,β-CTx,PTH,ALP,FBG,HbAlc,Creatinine and albumin/Cr in the osteoporosis group were significantly higher than controls(t=2.212,4.431,2.215,3.544,0.433,1.629,0.365,5.436,P＜0.01 or P＜0.05),the 25-(OH)VD3 in the osteoporosis group was lower than in controls(t=2.700,P＜0.01).Correlation analysis showed TP Ⅰ NP was negatively correlated with the BMD of femoral Troch and Ward's triangle (r=-0.413,-0.375,P＜0.01),and β CTx was negatively correlated with the BMD of femoral Troch (r=-0.301,P＜0.05).The TP Ⅰ NP,β-CTx in the overweight group and obesity group were higher than in normal group(F=50.59,51.28; F =96.20,95.71,all P＜0.01);the melbine group had higher TP Ⅰ NP,β-CTx than the thiazolidinedione and sulfonylurea group(F=32.33,33.35,F=31.07,39.18,P＜0.01);the TP Ⅰ NP,β-CTx in the better blood glucose control group were lower than in the bad control group(F=11.32,13.69,P＜0.01);the TP Ⅰ NP,β-CTx in the group with complication were increased as compared with no complication group(F=52.75,70.34,P＜0.01).Conclusions The spine BMD in elderly T2DM patients is decreased to some degree.The combined application of TP Ⅰ NP and β-CTx is helpful for the early diagnosis of osteoporosis in patients with type 2 diabetes.

Objective To evaluate the significance of tumor markers CEA and CA15-3,and biochemical markers of bone turnover (total procol]agen type Ⅰ amino-terminal propeptide (TP I NP),β-isomerized carboxyterminal propeptide (β-CTx),ALP and PTH) in the diagnosis of bone metastasis from breast cancer.Methods A total of 78 patients (all females) with mean age (56.72 ± 10.76) years,who were diagnosed with breast cancer,were included in this study.The patients were divided into two groups based on radionuclide bone imaging:with bone metastasis (n =32) and without bone metastasis (n =46).The serum concentrations of CEA,CA15-3,TP I NP,[β-CTx,PTH,ALP were measured.Gleason scores were evaluated.The diagnostic value was evaluated by ROC curve.The two groups were compared using two-sample t test.The correlations between bone metastasis and tumor markers,bone metastasis and biochemical markers of bone turnover were analyzed with Pearson correlation and logistic analysis.Results The serum levels of CEA,CA15-3,TPINP,β-CTx,PTH and ALP were significantly higher in the group with bone metastasis than those in the group without bone metastasis (t:4.16-7.56,all P ＜ 0.05).For the diagnosis of bone metastasis from breast cancer,the AUC of CEA,CA15-3,TPINP,[β-CTx,PTH and ALP was 0.815,0.887,0.869,0.852,0.844,0.731,respectively.Using the cut-off values of 4.18 μg/L for CEA,0.04 U/L for CA15-3,49.70 μg/L for TP I NP,0.47 pg/L for β-CTx,54.90 ng/L for PTH and 49.90 U/L for ALP,the sensitivities were 56.3％ (18/32),75.0％ (24/32),78.1％ (25/32),81.3％ (26/32),78.1％ (25/32),68.8％ (22/32) and the specificities were 80.4％ (37/46),84.8％ (39/46),76.1％ (35/46),78.3％ (36/46),69.6％ (32/46),58.7％ (27/46),respectively.CEA,CA15-3,TPINP,β-CTx,PTH,ALP and Gleason score were positively correlated with the presence of bone metastasis (r:0.267-0.636,all P ＜ 0.05).CEA,CA15-3,TP I NP,β-CTx,PTH and Gleason score were independent predictors for bone metastasis of breast cancer (odds ratios:2.45,3.44,1.24,1.54,1.11,2.22,all P ＜0.05).The total coincidence rate of regression model was 81.3％ (26/32) in patients with bone metastasis.Conclusions The diagnostic values of CEA,CA15-3,TP I NP,β-CTx and PTH are comparable.Combined use of these parameters may be helpful for the early diagnosis of bone metastasis from breast cancer.

Objective To investigate the relationship between the biochemical markers of bone turnover and renal osteodystrophy (ROD) in patients with primary nephrotic syndrome (PNS).Methods A total of 30 patients with PNS and 50 healthy subjects (controls) were included in the study.The BMD of lumbar vertebrae and femur was measured by dual-energy X-ray absorptiometry.The levels of total procollagen type Ⅰ amino-terminal propeptide (TP Ⅰ NP),β-isomerized carboxyterminal propeptide (β-CTx),intact PTH (iPTH),serum calcium,serum phosphorus,ALP,25-OH-Vitamin D3 (25-OH-VD3),β2-micro-globulin(β2-MG),and ratio of urinary to creatinine (UA/Cr) were measured and calculated.The risk factors related to ROD in PNS patients were analyzed.Two-sample t test,multiple linear regression and partial correlation analysis were used to analyze data.Results The BMD values of lumbar vertebrae and femur in the PNS group were significantly decreased compared with those in controls (t =6.162,5.583,3.891 (＜40 years),5.923,5.324,3.129 (≥40 years),all P＜0.05) and the serum levels of TPⅠNP,β-CTx,iPTH,ALP,β2-MG and UA/Cr in the PNS group were significantly higher than those in controls (t:2.738-10.129(＜40 years),3.226-12.581 (≥40 years),all P＜0.05),and the levels of serum calcium and 25-OH-VD3 in the PNS group were significantly lower than those in controls (t =3.624,7.223 (＜40 years),2.011,2.564 (≥40 years),all P＜0.05).But there was no significant difference for serum phosphorus between the 2 groups (t=0.811,0.513,both P＞0.05).TP Ⅰ NP was positively correlated with β-CTx,iPTH,ALP,UA/Cr,β2-MG(r:0.512-0.682,all P＜0.01),and TP Ⅰ NP was negatively correlated with serum calcium and 25-OH-VD3(r=-0.314,-0.562,both P＜0.01)in the PNS group.β-CTx was positively correlated with iPTH,ALP,UA./Cr,β2-MG(r:0.459-0.618,all P＜0.01),and negatively correlated with serum calcium and 25-OH-VD3(r=-0.212,-0.589,both P＜0.01).The iPTH was positively correlated with ALP,UA/Cr and β2-MG (r =0.410,0.606,0.508,all P＜0.05),and negatively correlated with serum calcium and 25-OH-VD3(r=-0.315,-0.516,both P＜0.05).Conclusions The BMD in PNS patients is lower than that in healthy subjects.Combined measurement of TP Ⅰ NP,β-CTx and the BMD is helpful for the diagnosis of ROD in PNS patients.

Objective To further investigate the accuracy of the real-time three-dimensional echocardiography(RT-3DE) in evaluating right ventricular(RV) systolic function.Methods RT-3DE,MRI and SPECT were used to study a total 30 patients including 10 with coronary artery disease,7 with hypertension,5 with cardiomyopathy,5 with atrial septal defect,and 3 with pulmonary hypertension.The RV end-diastolic volume(RVEDV),RV end-systolic volume(RVESV),RV stroke volume(RVSV) and RV ejection fraction(RVEF) were measured.The correlation and the difference between different modalities were compared.Results RT-3DE had a good correlation with MRI in RVEDV,RVESV,RVSV and RVEF (rEDV =0.811,rESV =0.837,rSV =0.818,and rEF =0.701).Also RT-3DE had a correlation with SPECT in RVEDV,RVESV,and RVSV(rEDV =0.526,rESV =0.493,rSV =0.514),but there was no correlation between RT-3DE and SPECT in RVEF (rEF =0.235).Conclusions RT-3DE can assess RV systolic function accurately.

Objective To explore the effects of graphene oxide (GO)-polyethylene glycol (PEG)-folic acid (FA)-pyrenemethylamine hydrochloride (PyNH2)-mediated RNA interference (RNAi) of hypoxia-inducible factor-1α (HIF-1α) on the biological behaviors of human pancreatic cancer Patu8988 cells.Methods GO-PEG-FA-PyNH2 and RNAi targeting HIF-1α gene (GO-PEG-FA-PyNH2-HIF-1α-RNAi)was constructed.The expressions of HIF-1α and glucose transporter 1 (Glut-l) in Patu8988 cells were determined after knockdown of HIF-1α by RNAi.The invasive ability,the proliferation and the cell cycle of Patu8988 cells were investigated.The effect of HIF-1α knockdown on the uptake of 18F-fluorodeoxyglucose (FDG) in Patu8988 cells was also detected.Comparison of data was conducted by one-way analysis of variance and least significant difference t test.Results The GO-PEG-FA-PyNH2 was successfully constructed,and no cytotoxicity was found.Under the hypoxia or normoxia state,the mRNA and protein levels of HIF-1α and mRNA level of Glut-1 in cells transfected with GO-PEG-FA-PyNH2-HIF-1α-RNAi (study group) were lower than those in cells transfected with GO-PEG-FA-PyNH2 (negative group) and cells transfected with Opti-minimal essential medium (Opti-MEM,control group;F=30.25-32.58,t=3.66-5.81,all P＜0.05);the numbers of migrated cells in the study group were much lower than those in the negative group and the control group (F=38.63 and 41.35,t=20.51-35.25,all P＜0.01);the cell proliferation in the study group was significantly lower than that in the negative group and the control group (F=35.19 and 38.11,t =15.11-22.19,all P＜0.05).The proportions of G0/G1 cells in the study group were higher than those in the negative group and the control group (F=34.60 and 34.83,t=11.55-34.56,all P＜0.05);the 18 F-FDG uptake in the study group was lower than that in the negative group and control group (F=29.85 and 31.69,t =3.35-4.35,all P＜0.05).Conclusion GO-PEG-FA-PyNH2-mediated HIF-1α RNAi inhibits the expression of HIF-1α in pancreatic cancer cells,leading to changes in related biological behaviors.

Whether direct manipulation of Parkinson's disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6-10 months), and thus provides a practical transgenic monkey model for future PD studies.
Animals ; Brain ; CRISPR-Cas Systems/genetics* ; Dependovirus/genetics* ; Haplorhini ; Phenotype ; Protein Kinases/genetics*

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.