Objective To discriminate the species and individual identification with mitochondrial DNA (mtD-NA) sequencing in forensic medicine practice. Methods The multiplex PCR of mtDNA loop - D high - variation region and cytochrome- b region were investigated. The PCR products were detected with silver- stain method,followed by analysis of the PCR products with fluorescence sequence technique. Results The presence of two bands (358bp,279bp ) indicated the samples were from human, while only one band of 358bp indicated nonhuman origin. The part of mitochondrial DNA loop - D high - variation region (15997 ~ 16236) from 131 unrelated individuals of Guangdong population were sequenced. In all of these samples there were 69 nucleotide variations and 67 haplo-types.There was 2.679 mutation sites on average per person. The polymorphism was 97.92% . Conclusion The methods described here are reliable and very useful in species and personal identification of degraded samples.

[Summary] 2015 AACE/ACE Clinical Practice Guidelines for Integrated Management of Diabetes are updated and revised on the basis of 2011 edition. The guidelines highlight the comprehensive consideration of micro- and macro-vascular risks rather than merely focusing on the strategies and approaches of glycemic control. The guidelines also emphasize individualized targets for weight loss and the management of hyperglycemia ,dyslipidemia and hypertension and that more attention should be paid to safety of individualized treatment other than efficacy.

Objective To investigate the image quality differences of clinical conventional contrast injection solutions and personalized injection program in 64 -slice spiral coronary CT angiography (CTA),thus to obtain the CTA optimum contrast agent injection protocol.Methods 64 patients who needed to receive CTA angiography,were randomly divided into observation group and control group,32 cases in each group.The control group underwent rou-tine clinical angiography contrast agent injection protocol,the observation group received personalized contrast injec-tion solutions,the image difference of the two groups was compared.Results Compared with the control group,the right ventricular (RV)and pulmonary CT value of the observation group increased significantly[(272.4 ±72.3)HU, (372.1 ±78.2)HU],the differences were statistically significant (t =3.827,3.921,all P <0.05).CT values of left ventricular (LV),ascending /descending aorta had no significant differences between the two groups (P >0.05). Compared with the control group,the advantages of septal imaging of the observation group was significant (control group 3 points in 20 cases,29 cases of the observation group),the difference was statistically significant (χ2 =16.023,P <0.05).Conclusion In CTA angiography,personalized contrast injection speed can effectively improve the enhancement degree of right heart,so that the right ventricular septal has clearer image,provides a great conven-ience for clinical diagnosis and prognosis,it is worth further clinical application.

Objective To discuss the different contrast injection rate on the 64 -slice spiral CT scan liver arterial phase reinforced effect.Methods 60 patients who received abdominal CT scan were selected from January 2014 to April 2015.According to the different conditions,they were divided into control group (30 cases)and liver cirrhosis group (30 cases).Then each group based on packet injection rate into 2.5 mL/s (control group 1,cirrhosis group 1)and 3.5 mL/s (control group 2,cirrhosis group 2).Select Smartprep technology contrast agent tracking all patients,when the abdominal aorta CT value ≥150 HU start scanning.The departure time,CT value of each group were recorded,abdominal aortic artery,simultaneous analysis of each component of image quality comparison.Results When the contrast agent injection rate of 3.5 mL/s,excellent rate of the control group (95.5%)compared with the contrast injection rate of 2.5 mL/s (91.7%)had no significant difference (χ2 =0.021,P =0.638);The cirrhosis good rate (94.7%)compared with the contrast agent injection rate of 2.5ml/s (72.7%)had obvious advantages, the difference was statistically significant (χ2 =5.233,P =0.032).When the contrast agent injection rate of 2.5mL/s, cirrhosis abdominal aorta average peak enhancement [(187.25 ±21.00)HU]and the control group [(195.35 ± 19.00)HU]had no significant difference (t =0.826,P =0.436).The average trigger time of the control group (21.68 ±1.93)s was significantly less than the cirrhosis group (25.13 ±2.13)s,the difference was statistically significant (t =2.064,P =0.047).When the contrast agent injection rate of 3.5 mL/s,the control group,the mean abdominal aortic peak enhancement [(247.82 ±39.00)HU]was significantly greater than the cirrhosis group [(223.81 ±35.00)HU],the difference was statistically significant (t =2.652,P =0.037).The average trigger time of the control group (18.62 ±1.36)s was significantly less than the cirrhosis group (24.57 ±0.92)s,the difference was statistically significant (t =3.362,P =0.033).Conclusion The greater the contrast injection rate,arterial peak at the same time it increases the peak time come to shorten.When the contrast agent injection rate consistent with the control group,the average trigger time is short,high average peak enhancement.Cirrhosis needs higher contrast injec-tion rate.

AIM: To study the hypoglycemic action of cortex moutan polysaccharide 2b (CMP 2b) and the its mechanism. METHODS: Some animal models in this study included the glucose induced hyperglycemic model in mice, the alloxan induced diabetic model in mice and rats, and the HCSS induced insulin resistance model in mice. Those models were used to investigate the effects of CMP 2b on blood glucose of normal and hyperglycemic model animals, on serum superoxide dismutase (SOD), serum insulin, glycated hemoglobin (GHb), and apolipoproteinA 1 (ApoA 1) in alloxan diabetic animals. RESULTS: CMP 2b significantly lowered the blood glucose in hyperglycemic mice and rats induced by glucose and alloxan. CMP 2b also raised the SOD and ApoA 1 level, decreased the GHb level in diabetic animals, and improved insulin resistance in mice. CONCLUSION: CMP 2b can control experimental hyperglycemia. Its mechanism is related to improving insulin sensitivity and enhancing the utilization of glucose in peripheral tissues.

OBJECTIVEThis study explores the effects of different fibrin glue combination modes on the survival, proliferation, and apoptosis of dental follicle cells (DFCs), as well as to evaluate the feasibility and effectiveness of fibrin glue as transplantation material.

METHODSThe membranes of surviving DFCs were marked using 3,3'-dioctadecyloxa carbocyanine perchlorate (DIO), and the cell number was counted by using ImageJ2x software. The apoptotic cells were marked with prodium iodide (PI).

RESULTSCompared with that of the 3D-2 and 2D-1 groups, the degradation speed of the 3D-1 group was the slowest. DFCs could survive and grow well in fibrinogen with a concentration of 15 mg · mL⁻¹ supplemented with thrombin with a concentration of 2 U · mL⁻¹. In particular, the 3D-1 combination mode was significantly conducive to cell proliferation and stretching.

CONCLUSIONFibrin glue can be used as an effective cell transplantation material. The different combination modes have certain effects on cell proliferation. The 3D-1 combination mode is more conducive to the survival and proliferation of DFCs than other modes.

Apoptosis ; Cell Proliferation ; Cell Survival ; Dental Sac ; cytology ; Fibrin Tissue Adhesive ; pharmacology ; Fibrinogen ; Humans ; Thrombin

Some grain sprouts herbs recorded in Chinese Pharmacopoeia 2010, including Oryzae Fructus Germinatus, Setariae Fructus Germinatus and Hordei Fructus Germinatus, together with their adulterants, were identified by ITS2 sequences. The genomic DNA were extracted, their ITS2 sequences were amplified, and purified PCR products were
sequenced. Sequences were assembled using the CodonCode Aligner. The genetic distances, variable sites and the neighbor-joining (NJ) phylogenetic tree were computed by MEGA 6.0 in accordance with the Kimura 2-Parameter (K2P) model. The results showed that their intraspecific genetic distances were all much lower than the interspecific ones of themselves and their adulterants. The NJ tree based on ITS2 sequence indicated that Oryza sativa, Setariaitalica, and Hordeum vulgar formed one monophyletic clade respectively, in addition any one of the grain sprouts herbs and its adulterants could be distinguished clearly. Therefore, ITS2 sequence is suitable to be as a barcode to identify the grain sprouts herbs and their adulterants.

BacKground:Fanconi anemia( FA)pathway as a DNA crossIink damage repair pathway pIays an important roIe in maintaining genome stabiIity. In recent years,FA pathway was wideIy studied in DNA damage and cancer pathogenesis. Aims:To investigate the interaction between RAD18 and FANCD2 protein in human coIorectaI cancer ceII Iine SW480. Methods:Antigen-antibody compIex was co-precipitated by immunoprecipitation. Expressions of rabbit anti-human FANCD2 and RAD18 protein in antigen-antibody compIexes were detected by Western bIotting. The pIasmids of GST-RAD18 and GST were transferred into BL21 ceIIs and induced to express the target proteins. TotaI proteins of the ceII was extracted and GST-beads were used to conjugate the GST-RAD18 protein,and then incubated with the SW480 ceII Iysates, and Western bIotting was performed with the addition of rabbit anti-human FANCD2 antibodies. Results:RAD18 protein was detected in the antigen-antibody compIex from immunoprecipitation by using anti-FANCD2 antibody,and FANCD2 protein was detected by using anti-RAD18 antibody. FANCD2 protein was aIso detected by using anti-GST-RAD18 antibody. GST-RAD18 protein used as bait protein couId capture the FANCD2 protein in SW480 ceIIs. Conclusions:There is an interaction between RAD18 and FANCD2 protein in SW480 ceIIs,and aIso an interaction between GST-RAD18 and FANCD2 protein.

Objective To observe the effects of progesterone on the mRNA and protein expressions of nerve growth factor(NGF) and brain-derived neurotrophic factor(BDNF) in the focal cerebral ischemia-reperfused rats,and to explore its mechanism of brain protection. Methods Totally 96 healthy male SD rats were randomly divided into sham surgery group,ischemic reperfusion group,vehicle-treated group and progesterone-treated group (n=24 for each).The model of focal brain ischemia-reperfusion injury was induced by middle cerebral artery occlusion (MCAO).After 2 h temporary MCAO,rats were subjected to reperfusion for 3 h,6 h,12 h and 24 h,respectively.The mRNA and protein expressions of NGF and BDNF were analyzed by real time-PCR and Western blot,respectively. Results In the injured cortex,the mRNA and protein expressions of NGF and BDNF in ischemic reperfusion group came to the peak at 6 h after reperfusion,and then gradually declined to the level of sham operation group at 24 h after reperfusion.In progesterone treatment group,BDNF and NGF mRNA and protein expressions reach the peak at 12 h after reperfusion,and were still higher at 24 h after reperfusion than in ischemic reperfusion group(P＜0.05). Conclusions Progesterone plays a protective role in focal cerebral ischemia-reperfusion by increasing the expressions of BDNF and NGF in rats.

OBJECTIVETo explore the molecule mechanism of Salidroside inducing directional differentiation of mouse mesenchymal stem cells (MSCs) into neuronal cells.

METHODSThe mouse multipotent mesenchymal precursor cell line (D1) was taken as the objective. Cultured MSCs were divided into the negative control group (complete culture solution), the positive control group (containing 1 mmol/L β-mercaptoethanol), the Salidroside induced group (20 mg/L Salidroside), and the blocked group (20 ng/ ml DKK1, a special inhibitor of Wnt/β-catenin signal pathway). All cells were inoculated in a 6-well plate (1 x 10(4) cells/cm2) and grouped for 24 h. The expression of p-catenin was detected by fluorescence Immunochemistry in the negative control group, the positive control group, and the Salidroside induced group. The expression of neuron-specific enolase (NSE), beta 3 class III tubulin (β-tubulin III), nuclear receptor related factor 1 (Nurr1), glial fibrillary acidic protein (GFAP) mRNA, Wnt3a, β-catenin, low-density lipoprotein receptor-related protein6 (LRP6), Axin mRNA were detected using reverse transcrip- tion PCR (RT-PCR). The expression of β-catenin and NSE protein were analyzed by Western blot in the negative control group, the positive control group, and the Salidroside induced group. Ca2+ chelating agents (EGTA), L-type Ca2+ channel blocker (Nifedpine), and IP3Ks special inhibitor (LY294002) were used to block Ca2+ signal pathway respectively. The expression of Wnt3a, LRP-6, Axin, glycogen syn- thase kinase (GSK-3), and β-catenin mRNA were detected by RT-PCR. The β-catenin protein expression was analyzed using Western blot.

RESULTSCompared with the positive control group, β-catenin protein was strong positively expressed; the expression of Wnt3a, β-catenin, LRP6, Axin, NSE, β-tubulin III, Nurr1 mRNA, and NSE protein were obviously up-regulated in the Salidroside induced group (P < 0.01). Compared with the positive control group and the Salidroside induced group, β-catenin, NSE, Nurr1, and β-tubulin III mRNA expression decreased; β-catenin and NSE protein expression were also down-regulated in the blocked group (P < 0.01). Compared with the Salidroside induced group, the expression of Wnt3a, LRP-6, β-catenin, and Axin mRNA were down-regulated in the Ca2+ signal blocked group and the salidroside induced group (P < 0.01, P < 0.05).

CONCLUSIONSalidroside affected directional differentia- tion of MSCs into neuronal cells through Wnt/β-catenin and Ca2+ signal pathway.

Animals ; Cell Differentiation ; drug effects ; Glucosides ; pharmacology ; Glycogen Synthase Kinase 3 ; Lipoproteins, LDL ; Low Density Lipoprotein Receptor-Related Protein-6 ; Mesenchymal Stromal Cells ; physiology ; Mice ; Neurons ; Phenols ; pharmacology ; Phosphopyruvate Hydratase ; RNA, Messenger ; Signal Transduction ; Wnt Signaling Pathway ; physiology ; beta Catenin ; metabolism