Objective The purpose of this study was to investigate the feasibility of short tandem repeat(STR) genotyping of cell free DNA in plasma for individual identification and paternity testing. Methods EDTA-Na2 DNA anti-coagulant blood samples were collected from 36 unrelated healthy volunteers,and both DNA in leukocytes and cell free DNA in plasma were extracted respectively using phenol-chloroform method. Target DNA in blood cells and plasma were amplified using regular STR typing and fluorescent multiplex STR assay separately,accordingly,the PCR products were analyzed by polyacrylamide gel electrophoresis and capillary electrophoresis. Results Using either normal PCR-STR or fluorescent multiplex STR assay,the consistent STR genotyping results were detected with similar efficiency for cell DNA and plasma DNA samples from the same individual. Conclusion Cell free DNA in plasma samples can be used as useful biological samples for STR genotyping,which can be applied to individual identification and paternity testing in forensic practice.

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 the risk factors of portal venous thrombosis ( PVST) following laparoscopic splenectomy ( LS) in cirrhotic patients with hypersplenism .Methods The clinical data of 62 patients with secondary hypersplenism due to cirrhotic portal hypertension admitted to the Clinical Medical College of Yangzhou University were retrospectively analyzed .These patients underwent LS from January 2013 to April 2014.Based on whether the patients had PVST on postoperative day (POD) 7 or not, the pa-tients were categorized into the non-PVST and PVST groups .Fifteen perioperative variables were analyzed in these two groups .Results On univariate analysis , age and portal vein diameter in the PVST group were significantly higher and velocity of portal blood flow in the PVST group were significantly lower than those in the non-PVST group, (all P<0.05).Bivariate correlation analysis performed for the relationship between these variables and occurrence of PVST on POD 7 showed age >50 years, portal vein diameter >13 mm and velocity of portal blood flow >18 cm/s were significantly correlated .Multivariate logistic regression showed that age >50 years and portal vein diameter >13 mm were significantly independent risk factors of PVST, and velocity of portal blood flow >18 cm/s was an independent protective factor .Conclusion Age>50 years and portal vein diameter >13 mm were independent risk factors , while velocity of portal blood flow >18 cm/s was an independent protective factor of postoperative PVST .

Objective: In order to solve the problem of moisture absorption for Biantong Capsule, the extract granule of Chinese medicine is covered with HPMC for Biantong Capsule. Methods: Taking moisture absorption rate as measure index, various facters effected the moisture absorption rate, and the optimal coating process can be selected with well disturbed uniform design and computer analog test. Results: Under the condition of 25℃, relative moisture 75% and 100% respectively, the moisture absorption rate is lower than uncovered granules, the critic relative moisture of covered granules is greater than that of uncovered granules. Conclusion: Chinese medicine of Biantong Capsule covered with HPCM can be served as a good anti moisture method.

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 construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase (COX) activity in mouse Sertoli cell line TM4. Methods The coding region of Cox7a2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. The PCR product was inserted into pEYFP-C1 vector with BamH I and EcoR I restriction site, and confirmed by DNA sequencing. The recombinant fusion protein vector was amplified by transforming into DH5a and transfected into TM4 cells. The protein expression was identified by Western blot. COX activity was measured by spectrophotometer 6, 12, 24 and 48 h after the transfection of recombinant vector into the TM4 cell line. Results The entire coding sequence of Cox7a2 was cloned with 252 bp length. Plasmid pEYFP-C1-Cox7a2 vector was constructed and the positive clones were verified by restriction enzymes digestion and DNA sequencing. The transfection efficiency of the TM4 cell line was about 70% and 37000 D fusion protein was obtained. The COX activities were (0.642±0.051), (0.542±0.049), (0.311±0.021) and (0.216±0.010) U/mg 6, 12, 24 and 48 h after the transfection of recombinant vector in the TM4 cell line. Meanwhile, the COX activities were (0.714±0.064) and (0.653±0.031) U/mg in non-tranfected and naked vector group respectively. Compared with the non-tranfected group, COX activity decreased significantly 12, 24 and 48 h after the transfection. Conclusions The recombint plasmid vector was successfully constructed. Cox7a2 gene has an inhibiting effect on COX activity and may play an important role in the regulation of COX activity in mouse Sertoli cell line TM4.

Objective To investigate the feasibility and safety,and short-term therapeutic effect of laparoscopic azygoportal disconnection without splenectomy for cirrhotic portal hypertension (PLT count ＞ 50 × 109/L).Methods Clinical data of 48 patients with bleeding portal hypertension and secondary hypersplenism (PLT count ＞ 50 × 109/L) undergoing laparoscopic splenectomy and azygoportal disconnection (LSD,n =26) vs.laparoscopic azygoportal disconnection (LD,n =22) between January 2014 and August 2015 were analyzed.Results Operative time (82 ± 29) min,intraoperative blood loss 20(10-50) ml,days of postoperative fever 0(0-3) d,rate of postoperative fever 10/22,postoperative hospital stay (7.0 ± 1.3) d,and WBC counts (3.8 ± 1.6) × 109/L,PLT counts 64 (49-88) × 109/L,and the incidence of portal vein thrombosis on POD 7 (14％),were significantly less in LD group than in LSD group [(180±41) min,80(20-500) ml,2(0-4) d,(22/26),(10.8 ±3.0) d,(9.1 ±3.1) × 109/L,156 (78-630) × 109/L,(42％)],(t =9.637,Z =-4.746,Z =-2.314,x2 =8.224,t =5.794,t =7.785,Z=-5.508,x2 =4.742,all P ＜ 0.05).Immune function was better in LD group than in splenectomy group at postoperative month 3.The serum proportion of CD4 + (58 ± 11) and the CD4 +/CD8 + ratio (1.9 ±0.7) at postoperative month 3 were significantly higher after LD than after LSD [(43 ± 14),(1.2 ± 0.9)],(t =-3.755,t =-2.509,all P ＜ 0.05).Conclusion Laparoscopic azygoportal disconnection without splenectomy is safe and effective for esophagogastric variceal hemorrhage and moderate hypersplenism (PLT ＞ 50 × 109/L) secondary to portal hypertension.

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.

[Objective]This study was designed to determine growth inhibition of diallyl trisulfide(DATS)in human prostate cancer cells by inducing apoptosis and further to investigate the mechanism underlying such effect.[Methods]Growth inhibition by DATS was estimated by the tetrazolium(MTr)assay.Apoptosis induction in DATS-treated cells was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with DAPI and flow cytometric analysis of cells with sub-G1 DNA content following staining with propidium iodide.Protein levels of apoptosis regulating proteins were determined using western blot.The activity of caspase-3 was measured using a colorimetric assay.[Result]DATS showed tumor growth inhibition in a time-and dose-dependent manner,IC_(50) of DATS was 14 μmol/L at 72 h.DATS evoked apoptosis as confirmed by cell morphology and by the analysis of flow cytometry.The expression of Bcl-2 and Bcl-xL,the apoptosis-suppressing proteins,was more down-regulated.The activity of caspase-3 was enhanced by DATS.[Conclusion]DATS inhibits growth of prostate cancer cells by inducing apoptosis in association with down-regulation of Bcl-2 and Bcl-xL and activation of caspase-3.

Objective To find an ideal animal model of acute respiratory distress syndrome (ARDS) through investigating the characteristics of three two-hit animal models of ARDS.Methods Forty-eight SD rats were randomly divided into 4 groups: Control group [2.5 mL/kg normal saline (NS) i.v.given at 0 min and 30 min];OA+OA group [0.5 mL/kg oleic acid (OA) i.v.given at 0 min and 30 min];LPS+LPS group [2.5 mg/kg lipopolysaccharide (LPS) i.v.given at 0 min and 30 min];and OA+LPS group [0.5 mL/kg OA i.v.given at 0 min and 2.5 mg/kg LPS,i.v.given at 30 min].The samples were collected at 5 h after the second drug injection.White blood cells count (WBC),polymorphonuclear leukocyte ratio (PMN%),total protein concentration,tumor necrosis factor α (TNF-α) level in bronchoalveolar lavage fluid (BALF),arterial blood gas analysis and lung wet-dry weight ratio (W/D) were measured,respectively.Pathological changes in the lung tissues were observed and histological scores were evaluated.Results Compared with those in the control group,PaCO2,WBC,PMN%,total protein concentration and TNF-α levels in BALF were significantly increased,while PaO2 was dramatically decreased (P<0.01) in the OA+OA,LPS+LPS and OA+LPS groups.The levels of protein concentration in BALF and lung W/D ratio in the OA+LPS group were significantly higher than these in the LPS+LPS group (P<0.05 for all),but had no statistically significant difference compared with these in the OA+OA group.The levels of WBC,PMN% and TNF-α in BALF in the OA+LPS group were significantly higher than those in the OA+OA group (P<0.05),but not significantly different from those in the LPS+LPS group.The most typical pathological changes and the highest pathological scores were found in the OA+LPS group.Conclusions All the three different methods including OA+OA,LPS+LPS,and OA+LPS can be used to establish “two-hit” animal models of acute respiratory distress syndrome.The “two-hit” animal model of acute respiratory distress syndrome induced by OA+LPS is more closer to clinical ARDS and is useful for studies on the pathophysiology of ARDS,and is an ideal “two-hit” animal model of ARDS.