Source identification of human biological materials in crime scene plays an important role in reconstructing the crime process. Searching specific genetic markers to identify the source of different human biological materials is the emphasis and difficulty of the research work of legal medical experts in recent years. This paper reviews the genetic markers which are used for identifying the source of human biological materials and studied widely, such as DNA methylation, mRNA, microRNA, microflora and protein, etc. By comparing the principles and methods of source identification of human biological materials using different kinds of genetic markers, different source of human biological material owns suitable marker types and can be identified by detecting single genetic marker or combined multiple genetic markers. Though there is no uniform standard and method for identifying the source of human biological materials in forensic laboratories at present, the research and development of a series of mature and reliable methods for distinguishing different human biological materials play the role as forensic evi-dence which will be the future development direction.

Objective:To investigate the alteration of delayed memory and its relationship with neurogenesis and angiogenesis in vascular dementia (VD) rats after moxibustion therapy. Methods: Two hundred adult male SPF Wistar rats were chosen for the experiment. Thirty-six rats were randomly selected as the sham operation group. Except for rats in the sham operation group (n=36), the others were made into VD models by bilateral common carotid arteries occlusion (BCCAo). After modeling, the 108 survived rats were randomly divided into 3 groups: a model group, a neural stem cells (NSCs) plus endothelial progenitor cells (EPCs) moxibustion group and a NSCs moxibustion group. Co-transplanted implant was transplanted into the rats in the NSCs plus EPCs moxibustion group, and the rats in the NSCs moxibustion group were transplanted by NSCs only. The NSCs plus EPCs moxibustion group and the NSCs moxibustion group received suspended moxibustion therapy at Baihui (GV 20), Dazhui (GV 14) and Shenting (GV 24), (each group was divided into 3 subgroups by the treatment course as 1, 2 and 3 courses). Every group was measured by Morris water maze to evaluate its delayed memory after 3 treatment courses and the rat’s brain was taken out after perfusion of 4% paraformaldehyde one day after 1, 2 and 3 treatment courses, respectively. Marker protein expression was detected by laser confocal microscope to analyze the effect on neurogenesis and angiogenesis. Results: VD rats showed delayed memory in Morris water maze test 3 d after ischemic injury. After 3 courses of moxibustion therapy, VD-induced delayed memory deficits were improved in the NSCs plus EPCs moxibustion group and the NSCs moxibustion group. The expressions of nestin, doublecortin (DCX) and CD34 increased significantly in the two moxibusiton groups after every treatment course (all P＜0.05), which might contribute to the neurogenesis and angiogenesis in hippocampus. In addition, compared with the rats in the NSCs moxibustion group, the expressions of nestin, DCX and CD34 increased significantly in the NSCs plus EPCs moxibustion group (P＜0.05). Conclusion: Moxibustion can reverse VD-induced delayed memory deficits, which may be related to the promotion of neurogenesis and angiogenesis.

The key technology of the device for the viviperception of the animal mesenteric microcirculation is to simulate the celiac environment in the device. The technical requirements of the device for microcirculation viviperception are that the observation box should be able to "keep warm, preserve moisture, continually perfuse, and fix the sample"; and the lighting should be "intense", "convergence", and "cool". After actual application, it was found that the newly designed and developed the device by research personnel of Wannan Medical College for the viviperception of the animal mesenteric microcirculation can meet the technical requirements, which is able to "keep warm, preserve moisture, continually perfuse, and fix the sample", and using LED lamp as the microscope light source is "intense", "convergence", and "cool". This device is ingenious and reasonable in design, stable in technology, convenient in operation, and competent in microcirculation viviperception. It solves the technical problem to simulate the celiac environment for mesenteric microcirculation viviperception. The device provides convenience to observe and study the microcirculation, which is worth to be applicated widely.

Objective To investigate the efficacy and security of cypher stent(sirolimus-eluting stent)in the treatment of old patients with coronary heart disease(CHD).Methods From November 2002 to May 2005,328 elderly CHD cases(age:60-86 years)were treated with 415 Cypher stents.Among the 328 patients,66 had ST-segment elevation of myocardial infarction,21 had non ST-segment elevation of myocardial infarction,149 had unstable angina and 92 had stable angina.As for lesion characteristics,diffuse disease was found in 91 case(26.1%),bifurcation lesions in 68 cases(19.6%),chronic total occlusion lesions in 56 cases(16.0%),in-stent restenosis in 14 cases and ostial lesions in 15 case.The immediate angiographic outcome,major cardiac event(MACE) and angiographic follow-up at 6 months were assessed.Results Stent implantation was successfully achieved in 99% patients with CHD.Acute and sub-acute stent thrombosis occurred in 2 patients,late stent thrombosis with AMI occurred in 2 patients,1 died during the 6 months follow-up.The MACE rate during hospitalization was 0.6% and 3.6% during 6 months follow-up.Angiographic follow-up in 84 patients at 6 months showed that in-stent restenosis rate(ISR)was 8.3%(restenosis within the stents was 2.4%).The target vessel revascularization(TLR)rate was 5.9%.Conclusions Cypher stent implantation in CHD is safe and effective,the ISR rate and TLR rate are significantly lower than those of bare metal stents.

OBJECTIVETo explore the influence of exogenous putrescine and cadaverine on pro-inflammatory factors in the peripheral blood of rabbits.

METHODSForty ordinary adult New Zealand rabbits were divided into saline, necrotic tissue homogenate (NTH), putrescine, and cadaverine groups according to the random number table, with 10 rabbits in each group. Saline, NTH, 10 g/L putrescine, and 10 g/L cadaverine were respectively peritoneally injected into rabbits of corresponding group in the amount of 1 mL/kg. The blood sample in the volume of 2 mL was collected from the central artery of rabbit ears before injection and at 2, 6, 12, 24, 30, 36, 48, 60 hours post injection (PIH). Contents of TNF-α, IL-1, and IL-6 in the serum were determined with enzyme-linked immunosorbent assay. Data were processed with repeated measurement data analysis of variance and Spearman correlation analysis, and cubic model curve was applied in curve fitting for the contents of inflammatory factors.

RESULTS(1) The serum contents of TNF-α, IL-1, and IL-6 were increased in NTH, putrescine, and cadaverine groups in different degrees at most post injection time points. There was no significant change in the concentrations of the three pro-inflammatory factors in saline group, and they were significantly lower than those of the other three groups at most post injection time points (with F values from 3.49 to 13.58, P values all below 0.05). The serum contents of TNF-α, IL-1, and IL-6 in putrescine group began to increase at PIH 2, 6, and 6, which was similar to the trend of NTH group, but the changes were delayed compared with those of cadaverine group(all at PIH 2). The peak values of TNF-α, IL-1, and IL-6 in putrescine group were respectively (339 ± 36), (518 ± 44), and (265.9 ± 33.5) pg/mL, which were significantly lower than those of cadaverine group [ (476 ± 86), (539 ± 22), and (309.4 ± 27.1) pg/mL], with F values respectively 5.11, 1.90, and 5.56, P values all below 0.05. (2) The period of time in which contents of TNF-α, IL-1, and IL-6 began to increase (PIH 3-4) and the peaking time of the three pro-inflammatory cytokines (PIH 18-30) in putrescine group appeared later than those of cadaverine group (PIH 2 and 12-30). The duration of peaking time of the three pro-inflammatory cytokines in putrescine group was shorter than that of cadaverine group (PIH 18-30 vs. PIH 12-30). The increasing period and the duration of peaking time of TNF-α, IL-1, and IL-6 in putrescine group were close to those of NTH group (PIH 3-5 and 18-30). The correlation coefficient test analysis showed that the trends of changes in contents of three pro-inflammatory cytokines in putrescine group were significantly correlated with those of NTH group (r(TNF-α) = 0.933, P < 0.01; r(IL-1) = 0.967, P < 0.01; r(IL-6) = 0.950, P < 0.01). The obvious correlation between cadaverine group and NTH group was only found in the contents of IL-1 and IL-6 (r(IL-1) = 0.913, P < 0.01; r(IL-6) = 0.883, P < 0.05).

CONCLUSIONSBoth exogenous putrescine and cadaverine can cause inflammatory reaction in rabbits. The trend of the inflammatory reaction induced by putrescine was similar with that by NTH, suggesting that putrescine may play a leading role in the inflammatory reaction induced by necrotic tissue decomposition.

Animals ; Cadaverine ; adverse effects ; Inflammation ; blood ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Necrosis ; blood ; Putrescine ; adverse effects ; Rabbits ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate the expressions of the active form of glycogen synthase kinase-3(GSK-3)-pGSK-3α/β (Tyr279/216) and its downstream moleculor X-linked inhibitor of apoptosis protein (XIAP) in cholangiocarcinoma and to analyze their correlation with clinicopathological and survival significance.

METHODSImmunohistoehemistry was used to detect the expressions of the active form of GSK-3- pGSK-3α/β (Tyr279/216) and its downstream moleculor XIAP proteins in 50 cholangiocarcinoma tissues and 20 normal bile duct tissues.

RESULTSThe positive rates of pGSK-3α/β (Tyr279/216) and XIAP were 62.0% and 68.0% in cholangiocarcinoma, and 10.0% and 25.0% in normal bile duct tissues, respectively. The intensity of pGSK-3α/β (Tyr279/216) and XIAP expressions in cholangiocarcinoma were significantly higher than that in the normal bile duct tissues (P < 0.001), and there was a significant correlation between pGSK-3α/β (Tyr279/216) and XIAP expressions (r = 0.544, P < 0.001). The expression of pGSK-3α/β(Tyr279/216) protein in cholangiocarcinoma was associated with TNM stage (P = 0.042), histological grade (P = 0.031), whereas the expression of XIAP protein in cholangiocarcinoma was correlated with CEA level (P = 0.006). Patients with positive expression of pGSK-3α/β (Tyr279/216) and XIAP demonstrate a significantly worse prognosis than that of patients with negative expression of pGSK-3α/β (Tyr279/216) and XIAP for overall survival (P = 0.002, P = 0.018). Multivariate survival analysis revealed that positive pGSK-3α/β (Tyr279/216) expression provided significant independent prognostic value for overall survival (P = 0.002).

CONCLUSIONSThe expressions of pGSK-3α/β(Tyr279/216) and XIAP proteins were significantly associated with the development and progression of cholangiocarcinoma. pGSK-3α/β(Tyr279/216) may be an important prognostic factor for survival of patients with cholangiocarcinoma.

Bile Duct Neoplasms ; metabolism ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Carcinoembryonic Antigen ; blood ; Cholangiocarcinoma ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Prognosis ; Survival Rate ; X-Linked Inhibitor of Apoptosis Protein ; metabolism

OBJECTIVE: To study the changes of HSP 70 mRNA and c-fos mRNA expression and to find a method to differentiate antemortem from postmortem electrocution. METHODS: Fifteen New Zealand rabbits were randomly divided into three groups, the antemortem electrocution group, the postmortem electrocution group, and the control group. Each group consists of five rabbits. The levels of HSP 70 mRNA and c-fos mRNA in skeletal muscle and cardiac muscle were examined with quantitative fluorescent RT-PCR. RESULTS: The levels of HSP 70 mRNA and c-fos mRNA in the antemortem electrocution group increased significantly (P<0.05), compared with that of the postmortem electrocution group. CONCLUSION The changes of HSP 70 mRNA and c-fos mRNA expression in skeletal muscle and cardiac muscle can be used as an indicator to distinguish antemortem from postmortem electrocution.
Animals ; Electric Injuries/metabolism* ; Forensic Pathology ; HSP70 Heat-Shock Proteins/metabolism* ; Male ; Muscle, Skeletal/metabolism* ; Myocardium/metabolism* ; Postmortem Changes ; Proto-Oncogene Proteins c-fos/metabolism* ; RNA, Messenger/metabolism* ; Rabbits ; Random Allocation

OBJECTIVETo explore the effect of nitric oxide (NO) on human sperm capacitation and acrosome reaction (AR).

METHODSDifferent concentrations of sodium nitroprusside (SNP) were added to the sperm suspension from 48 healthy fertile men, and the suspension was incubated in 1 x Earle at 37 degrees C for 1 hour. Progesterone was used to induce AR for 15, 30, 45 and 60 min, and then acid phosphatase (ACP) activity in the suspension before and after capacitation and at different time of AR was measured by p-nitrophenyl sodium phosphate assay. In the meantime, sperm motile parameters were assayed by CASA to observe sperm capacitation and AR.

RESULTSACP activity and sperm motile parameters increased in the 50 approximately 100 nmol/L NO concentration group, showed no significant variation in the 150 approximately 200 nmol/L group, and decreased in the 250 approximately 300 nmol/L group.

CONCLUSIONNO can facilitate sperm capacitation, AR and sperm motile parameters in low concentration and suppress them in high concentration. ACP activity assay of sperm is an objective and reliable method to evaluate sperm capacitation and AR in whole sperm population.

Acid Phosphatase ; metabolism ; Acrosome Reaction ; drug effects ; physiology ; Adult ; Dose-Response Relationship, Drug ; Humans ; Male ; Nitric Oxide ; physiology ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Sperm Capacitation ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; enzymology

OBJECTIVETo investigate the relationship between mTOR signaling pathway and ALK-positive lymphoid cell lines.

METHODSThe expression of the downstream effector proteins of mTOR were analyzed by Western blot before and after Karpas299, BaF3/NPM-ALK and BaF3 cell lines treated with rapamycin. Effect of rapamycin on cell proliferation was detected by MTT assay. FACS was used to analyze apoptosis and cell cycles.

RESULTSmTOR signaling phosphoproteins, p-p70S6K and p-4E-BP1 were highly expressed in ALK(+) Karpas299, BaF3/NPM-ALK and parental BaF3 cell lines, and they were dephosphorylated after 1 h withdrawal of IL-3 in BaF3 cells. After 48 h exposure to 10 nmol/L rapamycin, p-p70S6K and p-4E-BP1 proteins expression were decreased, and mainly for the former. The relative inhibitory rate to its control cells was 24.4% in Karpas299, 37.8% in BaF3/NPM-ALK and 61.6% in BaF3. The apoptotic ratio was increased from (11.97 +/- 0.11)% to (15.87 +/- 0.62)% in Karpas299 (P < 0.05), from (3.23 +/- 0.11)% to (7.67 +/- 0.49)% in BaF3 (P < 0.05) and from (1.90 +/- 0.47)% to (2.80 +/- 0.27)% in BaF3/NPM-ALK (P > 0.05). The fraction of G(1) phase cells increased from (37.63 +/- 1.91)% to (69.77 +/- 5.44)% in BaF3/NPM-ALK, from (31.13 +/- 2.51)% to (40.70 +/- 1.47)% in Karpas299 and (53.57 +/- 2.22)% to (63.70 +/- 1.20)% in BaF3 (P < 0.05).

CONCLUSIONNPM-ALK kinase can activate mTOR signaling pathway. Rapamycin can inhibit the proliferation of ALK(+) lymphoid cells by blocking mTOR signaling pathway and inducing cell cycling arrest at G(1) phase.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Lymphoma ; metabolism ; pathology ; Mice ; Protein-Serine-Threonine Kinases ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Receptor Protein-Tyrosine Kinases ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; drug effects ; Sirolimus ; pharmacology ; TOR Serine-Threonine Kinases

OBJECTIVETo study the tissue culture and rapid-proliferation techniques of Pueraria mirifica.

METHODThe tender branch were used as explants and cultivated in different media. The optimum media for inducing buds, proliferation and rooting were selected by adjusting the kinds and doses of plant hormones and special compounds.

RESULTThe medium of MS + IBA 0.05 mg L(-1) + BA 0.5 mg L(-1) was suitable for buds inducing and could be used in the first generation cultivation; MS + IBA 0. 02 mg L(-1) + BA 0.2 mg L(-1) and MS +BA 0.1 mg L(-1) were employed by turns in subculture, 25 days propagation coefficient was 3.0; and the medium of 1/2MS + IBA 0.1 mg L(-1) + IAA 0.2 mg L(-1) + C (special compound) 10 mg L(-1) was used for roots inducing, the rooting rate was 76.9%. Rooting plantlets were transplanted in spring and summer; the surviving rate was 81.0%.

CONCLUSIONThis technique system could be employed for rapid propagation of P. mirifica.

Pueraria ; growth & development ; Tissue Culture Techniques ; methods