ObjectiveConstruction and identification of wild-type and mutant human o-synuclein (SNCA) gene lentiviral expression vector, and its stable transfection into the rat pheochromocytoma cells.MethodsThe genes were synthesized with particular primer, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU ( with green fluorescent protein (GFP) gene) to construct a lentiviral vector expression plasmid pGC-FU-SNCA-GFP and pGC-FU-SNCA-GFP.After digestion and sequencing,pGC-FU-SNCA-GFP,pGC-FU-SNCA-GFP plasmidandpackaging plasmidpHelper1. 0,pHelper2. 0 were co-transfected into rat pheochromocytoma cells (PC12 cells).A stable transfection was established in the PC12 cells. Results By detecting the level of tagged protein of GFP and the target protein, the pGC-FU-SNCA-GFP and pGC-FU-SNCA-GFP expression in target cells was verified. MTT assay was employed to detect cell apoptosis in lentiviral pGC-FU-SNCA-GFP transfected group, lentiviral pGC-FU-SNCAmu-GFP transfected group ( experimental groups), without virus group ( control group) and empty vector group( total four groups cells). After transfection, at different timepoints ( 1, 2, 3, 4 and 5 d) the proliferation of cells was slowed ( the F value was 4. 534, 196. 285, 411. 829, 1282. 049, 3135. 559, all P ＜0.05). PI single staining was used to examine the cell cycle. The percentages of G1 phase, G2 phase,M phase cells were all statistically significant ( the F value was 885.79, 45.03,207.11 ,all P ＜0. 05). The percentage of G1 phase cells in the four groups cells increased significantly (CON group:59. 10 ±0. 35, NC group:68.24 ±0.60, OE group:71.73 ±0. 11, OE group:74.66 ±0.35). Conclusion This study constructs a foundation for further investigation on the basic function of SNCA and apoptosis related diseases.

BACKGROUND: The biomechanical studies about calcium sulfate cement vertebroplasty are only limited to the single fracture vertebra, not performed in spinal compression fracture unit with posterior transpedicular screw fixation. Furthermore, performing experimental study in the whole function spine unit (FSU) conforms to actual clinical situation. OBJECTIVE: To assess the biomechanical properties of calcium sulfate vertebroplasty combined with posterior transpedicular screw fixation in treatment of thoracolumbar compression fractures. DESIGN, TIME AND SETTING: A controlled experiment was performed at the Biomechanical Laboratory of Shanghai University in March 2009. MATERIALS: Fifteen fresh thoracolumbar spines were harvested from male calves and made into T_(11)-L_1 FSU, then divided into 3 groups randomly: normal control group, posterior transpedicular screw fixation group and transpedicular screw fixation plus vertebroplasty group. METHODS: T_(12) flexion-compression fracture models were made in all specimens of posterior transpedicular screw fixation group and transpedicular screw fixation plus vertebroplasty group, undergoing reduction and posterior transpedicular screw fixation, and calcium sulfate vertebroplasty combined with posterior transpedicular screw fixation respectively. MAIN OUTCOME MEASURES: All specimens were placed on the WE-10A universal testing machine for mechanical test. Load-straining, load-displacing, rigidity, strength and torsion of the FSU were performed in axial compression, flexion, extension and lateral bending states. The experimental outcomes were collected and compared by statistic analysis. RESULTS: The load-strain and loed-displacement showed a linear relationship. Straining values in vertebral body and intervertebral disc of calcium sulfate vertebroplasty plus posterior transpedicular screw fixation group were 14% and 12% less than that of posterior transpedicular screw fixation group, 21% and 13% less than that of normal control group. The thoracolumbar displacement in calcium sulfate vertebroplasty Plus posterior transpedicular screw fixation group decreased 25% and 37% as compared with other 2 groups respectively. Compared with normal control and posterior transpedicular screw fixation group, the thoracolumbar stiffness in calcium sulfate vertebroplasty plus posterior transpedicular screw fixation group increased 53% and 44% respectively. The strength in vertebral body and intervertebral disc of calcium sulfate vertebroplasty plus posterior transpedicular screw fixation group were 14% and 24% higher than that of posterior transpedicular screw fixation group, 13% and 20% higher than that of normal control group. The maximal twisting strength of FSU in calcium sulfate vertebroplasty plus posterior transpedicular screw fixation group were 18% and 30% higher than that of other 2 groups, the twisting stiffness were 30% and 40% higher than that of other 2 groups. The data above were significant differences statistically (P < 0.05). CONCLUSION: Posterior transpedicular screw fixation combined with calcium sulfate vertebroplasty show superior biomechanical properties for treatment of thoracolumbar compression fractures, which exhibits not only strong strength and stiffness, but also stable FSU, thus could decrease the stress loading of the internal fixation, the incidences of screw breakage and avoid the altitude loss of vertebral body.

Objective To investigate the relationship between serum homocysteine and post-stroke depression in elderly. Methods 62 elderly first stroke patients with post-stroke depression, 20 first stroke patients without depression, and 100 healthy elderly as control were detected their fasting serum homocysteine with fluorescence polarization immunoassay. Results Serum homocysteine were (25.59±9.47) μmol/ L, (18.56±7.84) μmol/L, and (12.67±6.45) μmol/L in the depression group, non depression group and healthy group, respectively (P<0.05). The level of homocysteine correlated with the score of Hamilton Depression Scale (r=0.8953, P<0.05) and age (r=0.8462, P<0.05). Conclusion The level of serum homocyteine might be one of risk factor of depression in patients with stroke.

Objective To investigate alterations of balance function in patients with mild-moderate Alzheimer's disease (AD) and with amnestic mild cognitive impairment (aMCI),and the possibility of using posturography to differentiate aMCI,mild-moderate AD and normal subjects. Methods The balance function of 20 patients with mild-moderate AD and 20 patients with aMCI were evaluated by posturography,and 20 healthy subjects of the same age range were recruited as controls.Results All posturography measures were significantly altered in mild-moderate AD patients compared with normal controls,with limits of stability( ( 15 398 ± 926 ) mm2 vs ( 31 654 ± 2132 ) mm2 ),open-eyed Mean X ( ( 10. 2 ± 4. 1 ) mm vs (5.8 ± 1. 4)mm) ,Mean Y(( -29.8 ± 10.2)mm vs ( -14.9 ±4.4) mm),Max X((30.5 ±9.5)mm vs (18.3 ±4. 1)mm ),Max Y((42.7 ± 11.4)mm vs (23.3 ±6.8)mm),LSKG((528.4 ± 105.4)mm vs (390. 3 ± 68.4 ) mm ),SSKG ( ( 252. 5 ± 89. 7 ) mm2 vs ( 178.8 ± 40. 9 ) mm2 ),close-eyed Mean X ((13. 1 ±4. 5) mm vs (7.9 ± 1.5)mm) ,Mean Y (( -58.2 ± 16. 9) mm vs ( -25.6 ±5.4) mm) ,Max X ((37.7±10.5)mm vs (24.7 ±7.3) mm ),Max Y ((78.5±18.7)mm vs (39.9 ±9.9) mm),LSKG ((816.6±171.3) mm vs (533.5 ±97.4) mm),SSKG((649.0 ± 129.7) mm2 vs (290.5 ±73.3) mm2),respectively ( t = 8.57; open-eyed F = 17.41,38. 10,60. 46,102. 10,29. 31,27. 85; close-eyed F = 37.20,541.79,34. 51,185.56,122. 83,384. 27 ;all P ＜0. 05) ;limits of stability ( (23 921 ± 1637 )mm2 vs (31 654 ±2132 ) mm2 ) and mean Y ( Antero-posterior sway,( - 39. 8 ± 8. 6 ) mm vs ( - 25.6±5.4 ) mm) were the only parameters which discriminated between aMCI and normal controls,respectively ( t = 6. 50,P = 0. 038; t =- 15.34,P = 0. 012). Conclusions Impairment in balance is a feature not only of mild-moderate AD,but also of aMCI,and posturography may be used as a possible test in differentiating between normal subjects,patients with aMCI and patients with mild-moderate AD whose motor performance and balance features are otherwise clinically normal,limits of stability and mean Y are the most sensitive parameters.

Epigenetics is the study of heritable changes in gene expression other than the changes in the underlying DNA sequence. Such changes include DNA methylation,genomic imprinting,X chromosome inactivation and non-coding RNA regulation. Recent progresses on epigenetics offer new ideas to tackling these problems in forensic science,including determination of the necessary allele in paternity,identification of fetal paternity testing in embryonic period,discrimination of identical twins,origination analysis of tissue,and individual age estimation. This review focuses on the main concept of epigenetics and its application in the field of forensic science.

Objective To investigate the safety and clinical efficacies of synchronous laparoscopic hepatectomy combined with splenectomy for the treatment of hepatocellular carcinoma (HCC) associated with cirrhotic portal hypertensive hypersplenism.Methods The clinical data of 5 patients with HCC associated with cirrhotic portal hypertensive hypersplenism who were admitted to the Clinical Medical College of Yangzhou University between January 2015 and April 2015 were retrospectively analyzed.Five patients underwent synchronous laparoscopic hepatectomy combined with splenectomy and intraoperative autologous blood transfusion.The operation time,volume of intraoperative blood loss,time for diet intake,postoperative drainage tube removal time,duration of hospital stay and occurrence of complications were observed.Patients were followed up by outpatient examination and telephone interview till June 2015.Measurement data with normal distribution were presented as x ± s.Results Of the 5 patients without conversion to open surgery,3 patients underwent synchronous laparoscopic hepatectomy combined with splenectomy,1 patient underwent laparoscopic hepatectomy + splenectomy + cholecystectomy and 1 patient underwent laparoscopic hepatectomy + splenectomy + devascularization.The operation time and volume of blood loss were (225 ± 41)minutes and (221 ± 81)mL.All the patients received intraoperative autologous blood transfusion without homologous blood transfusion.Two patients were complicated with abdominal effusion including 1 patient associated with pleural effusion,and symptoms of 2 patients disappeared after conservative treatment.All the patients took fluid diet at postoperative day 1 and out-of-bed activity at postoperative day 2,and drainage tubes were removed at postoperative day 3,with good recovery and without perioperative death.The duration of postoperative hospital stay was (7.8 ±0.8)days.All the 5 patients were followed up and survived well up to the end of follow-up.Conclusion Synchronous laparoscopic hepatectomy combined with splenectomy is safe and feasible for the treatment of HCC associated with cirrhotic portal hypertensive hypersplenism with an exact curative effect.

Objective Exogenous small-diameter vascular grafts have been developed with tissue-engineered small-di-ameter vascular grafts for the reconstructive surgery in the treatment of patients with coronary artery diseases in whom restenosis of the initially transplanted autografts occurred.This study was conducted to develop decellularized umbilical arteries (HUAs) and evaluate their physical and mechanical properties,as well as to assess the usefulness of decellularized HUAs in the coronary artery bypass grafting procedures.Methods After HUAs were harvested,their inner diameters were measured with a sliding caliper and the blasting pressure was measured with a pressure gauge.HUAs were firstly infused with a compound liquid consisted of 0.25％ Trypsin and 0.01％ Ethylene Diamine tetraacetic Acid(EDTA) and then with 1％ Sodium Lauryal Sulfate (SDS).Fragments of the vessels were collected and observed under light microscope and electron microscope.The mechanical characteristics of HUAs were identified with an electronic experiment machine before and after decellularization.Fibroblasts and endothelial progenitor cells growing along the lining of decelluarized human umbilical arteries(dHUAs) could be observed.Results The average inner diameter of HUAs was ( 3.50 ± 0.55 ) mm.Two of thirty HUAs were broken at a pressure of 300 mm Hg.The process for the lining of HUAs to be decellularized with a peristaltic pump took 0.5 hour with 0.25％ Trypsin and 0.01％ EDTA and 3 hours with 1％ SDS.Observations with light microscope and electron microscope demonstrated that all of the original lining cells were decellularized.The ultimate stress of the HUAs did not change significantly after decellularization ( P ＞0.05 ).Attached fibroblasts and endothelial progenitor cells could be seen along the decellular lining of the grafts and the ultimate stress of the HUAs did not change significantly after implantation of the two kinds of cells.Conclusion dHUAs,with good histocompatibility and properties mentioned above,may be used as potential vascular grafts in CABG.

Objective To investigate the influence of cartilage degeneration grades of patellar on the clinical outcomes after total knee arthroplasty (TKA) without patellar resurfacing.Methods 151 patients performed TKA without patellar resurfacing from February 2007 to January 2010 were retrospective studied.There were 65 males and 86 females,with the mean age of 65±5.0 years (range,56-82 years).The mean duration of follow-up was 3.5 years (range,2-5 years).Intraoperatively,the cartilage degeneration was classified according to Outerbridge classification.At the final follow-up,the patients' satisfaction was evaluated,and the incidence of anterior knee pain was assessed using visual analogue scale (VAS).If VAS was more than 3 points,patient was defined suffered anterior knee pain.Compare KSS and patella score of patients with different grades of cartilage degeneration.Results The articular cartilage of the patella was graded according to Outerbridge:grade Ⅰ in 18 patients,grade Ⅱ in 36 patients,grade Ⅲ in 62 patients,and grade Ⅳ in 35 patients.At the final follow-up,the patients' satisfaction rate and the anterior knee pain rate was 96.7％ 046/151) and 4.0％ (6/151),respectively.In the patients with different grades of cartilage degeneration,the patient satisfaction rate (H=5.54,P=0.14),anterior knee pain rate (H=0.56,P=0.91),KSS (knee score:F=1.95,P=0.12； function score:F=2.11,P=0.10) and patella score (F=1.35,P=0.26) were not found to be significantly difference.Conclusion The differentiae grades of cartilage degeneration may not affect the clinical outcomes of TKA.TKA without patella resurfacing appears to be a reasonable option for degenerative osteoarthritis.

OBJECTIVETo evaluate the clinical value of T lymphocyte subsets in prediction of chemotherapy responses of patients with pulmonary adenocarcinoma.

METHODSFifty-five chemotherapy-naive patients with pathologically or cytologically confirmed pulmonary adenocarcinoma were examined for peripheral blood T lymphocyte subsets using flow cytometry, including CD3(+) T cells, CD3(+)CD4(+) T cells, CD3(+)CD8(+) T cells, CD45RO(+) T cells and CD45RA(+) T cells.

RESULTSPatients who responded favorably to chemotherapy (CR(+)PR) showed a significantly lower percentage of CD45RA(+) T cells than those who failed to respond to chemotherapy (P=0.04). CD45RO(+) T cell percentage were slightly higher in the response group than in the non-response group, but this difference was not statistically significant (P=0.25). The other T cell subsets, namely CD3(+), CD3(+)CD4(+), and CD3(+)CD8(+) T cells showed no significant differences between the two groups.

CONCLUSIONA high percentage of peripheral blood CD45RA(+) T cells is associated with a poor short-term outcome of chemotherapy in patients with advanced pulmonary adenocarcinoma. Peripheral blood CD45RA(+) T cell level can be a reliable index for predicting chemotherapy efficacy in these patients.

Adenocarcinoma ; blood ; immunology ; Adult ; Aged ; Female ; Humans ; Leukocyte Common Antigens ; metabolism ; Lung Neoplasms ; blood ; immunology ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

Objective To genotype mixed samples with next generation sequencing and evaluate its prospects in forensic DNA application. Methods Three mixed biological samples from rapes cases and their reference samples were collected. DNA was extracted using the MagAttract M48 DNA Manual Kit(200). The ForenSeqTMDNA Signature Prep Kit was used for library preparation, and next generation sequencing was performed on the MiSeq FGx system. The ForenSeqTMUniversal Analysis v1.2.1 software was used for data analysis. NGS-based STR results were compared with CE-based genotypes. Results A single length polymorphic STR allele in the mixed profile could be recognized as two sequence polymorphic STR alleles from different donors, which would assist mixed profile analysis. Such phenomenon was observed in D3S1358, D9S1122 and D13S317 in this work. Conclusion Our results suggested that precision STR genotyping of mixed samples based on NGS can provide more information and hints for mixed STR profile separation.