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AIM: To study the expression of c-fos mRNA in brain following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expression following percussion. METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury group. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa). The injury groups were then subdivided into 5 min, 15 min, 30 min, 1h, 2h groups according to the time elapsed after injury. The expression of c-fos mRNA was studied with reverse transcription polymerase chain reaction (RT-PCR) semi-quantitatively.RESULTS: At 5 min after percussion, the induction of c-fos mRNA was increased, and remained elevated up to 2 h after brain injury.CONCLUSION: The induction and expression of the c-fos mRNA in cortex and brain stem after fluid percussion brain injury were increased rapidly.
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AIM: To study the expression of c-jun in brain stem following moderate lateral fluid percussion brain injury in rats, and to observe the temporal patterns of its expressions following percussion.METHODS: Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of injury group subjected to moderate lateral fluid percussion injury (0.2 mPa), and then were subdivided into 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 12 h groups according to the time elapsed after injury. The expression of c-jun was studied by immunohistochemistry and in situ hybridization. RESULTS: After percussion for 15 min, Jun positive neurons increased in brain stem progressively, and peaked at 12h. At 5min after percussion, the induction of c-jun mRNA was increased, and remained elevated up to 1h-2h after brain injury. CONCLUSION: The induction and expression of the c-jun in brain stem after fluid percussion brain injury were increased rapidly and lasted for a long time.
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Anti-GC serum was successfully prepared in two New Zealand rabbits immunized with GC protein which was isolated and purified from GC2-2 serum previously in our laboratory. The results of identification showed that the specificity of the home made anti- GC serum and the commercial anti- GC serum (DAKOPATTS) were identical. The titer of the home made anti-GC serum was 128. Three common phenotypes, GC1-1, GC2-1 and GC2-2could be identified by immunoelectrophoresis with the home made anti-GC serum. The concentration of GC protein as low as 3.1 ?g/ml could be detected by double immunodiffusion. In addition the anti-GC serum does not cross react with other human serum proteins.
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The C3 cleavages of human blood kept at different temperatures in vitro were studied. In 14 autopsy cases, the postmortem intervals(PMI) were estimated based on the C3 cleavage rates and the rectal temperatures. The 95% confidence of predicting PMI was?11 hours. The phenotyping of C3 could be achieved while the C3 cleavage rate was measured. It is suggested that it is useful for the personal identification of unknown corpses.
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Simultaneous phenotyping of AHSG, Pi and GC by IEF is reported. The results showed that the cumulative discrimination power and the cumulative exclusion probability of paternity of this method were 0.9701 and 0.58.11 respectively. It was proved to be the most efficient method for individual identification among the simultaneous phenotypings of genetic markers.It has been applied to paternity test and the results were satisfactory.
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This paper reports the detection of G2m(n)factors in human sera using theenzyme-linked immunosorbent inhibition test with monoclonal antibodies agai-nst G2m(n)factor(SH-21).The gene freguency of G2m(n)factor among 517unrelated individuals of ban population in Chengeu area was 0.5493 and itsvariance was 0.0004.
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The distribution of C_3 phenotype frequencies in the Han population in Cheng-du area was studied by means of cellulose acetate electrophoresis followed byimmunofixation.In 400 unrelated healthy individuals three C_3 phenotypes weredemonstrated.Their frequencies were as follows:SS=397,FS=2 and SSvar=1.Their gene frequencies were as follows:C_3~S=0.9963,C_3~F=0.0025 and C_3~(svar)=0.0013.The C_3 phenotype frequencies were in good agreement with those expec-ted.C_3 phenotyping in the huma bloodstains kept at 37℃ and room temp-lasserature for two days,at 4℃ for 23 days(cotton bloodstains)and for 35 days(g-bloodstains),and -20℃ for at least 87 days could be performed.C_3 ph-enotypingin human serum could be performed at roon temperature for 3 days,as 4℃ for 13days and at -20℃ for at least 106 days.C_3 phenotyping was usedin five casesof paternity dispute.
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Objective Study on the pattern of changes of bFGF and FGFRl mRNA occurred in the experimental brain injury model in order to provide scientific basis for the diagnosis, forensic identification and clinical treatment, and also for further ascertaining the molecular mechanism of brain injury. Methods Male Sprague-Dawley rats were divided into 3 groups: normal control, sham operation, and injury groups. The rats of injury groups were subjected to moderate lateral fluid percussion brain injury (0.2MPa). The injury groups was then subdivided into 30min, 1h, 3h, 6h, 12h, 1d, 3d and 7d groups according to the time elapsed after injury. In situ hybridization (ISH) and RT-PCR were used for studying the mRNA expression of both bFGF and FGFRl factors. Results (1) In the brain of normal control and sham operation control groups, mRNA levels of bFGF and FGFRl were low; (2) There is gradual increase of bFGF and FGFRl mRNA levels could be observed 6h to 3d after injury both in cortex and brain stem, then partly declined at 7d; (3) In hippocampus, the gradual increase occurred during 3h- 1d after injury, then partly declined at 3d, and returned to basal level at 7d. Conclusions The results suggested that brain injury induced the gene expressions of bFGF and FGFR1. The bFGF may contribute to maintenance of nerve cell survival and the repair of damaged neural tissues after CNS injury and the patterns of their level change were quite regular. It is potentially useful for timing of injury in forensic medical practice.
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?-1 acid glycoprotein, also named orosomucoid (ORM), is one kind of serum protein with genetic polymorPhism. Anti-ORM serum is necessary to phenotyping ORM. This communication describes the preparation of the antiORM serum. The anti-ORM sera were produced in three New Zealand rabbits cimmunized with ORM which was isolated and purified from human sera previously in our laboratory. The results of identification showed that the specificity of the home made anti-ORM and the commercial anti-ORM sera (Sigma) were identical. The titer of the home made anti-ORM serum was 128. 2.4?g/ml ORM could be detected by double immunodiffusion with the anti-ORM serum. In addition, the anti-ORM serum did not cross-react with other human serum proteins.
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This paper reports the determination of sex of human blood stains in threecriminal cases by the DNA amplification technique.The results fit the cases de-tails and gave the scientific evidence for the justice.The blood stains were dige-sted with proteinase K.The protein was extracted with the phenol-chloroform.The DNA was precipitated with NaCl and ethyl alcohol.Amplification of DNAwas carried out using two pairs of primer Y1.1 Y1.2 and Alu9.1 Alug.2 and heatstable FD polymerase.The amplified products were subjected to agarose gel(con-taining ethidium bromide)electrophoresis.Sex of blood stains was determinedaccording to the amplified products.PCR technique is rapid to perform,sensitiveand simple.No special equipments and isotope labelled probe are required.
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Identification of the human semen stain is reported using the latex particle agg-lutination inhibition test.The latex particles were coated with the human seminalplasma.The raw antihuman semen sera were previously absorbed with the humancolostrum.The results indicated that this method was more sensitive and acc-urate than sperm detection method.The sensitivty and the accuracy of the latex-particle agglutination inhibition test is the same as those of the double immunodiffusion test-using anti-p30 serum.Moreover,this method is easy to porform,nottime consuming and more practical.
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The distribution of ABH substances in human tissue cells was studied using the specific red cell adherence test(SRCA test).Tissues were taken from 11 cadavers of known ABO type and secrete status,fixed with 10% neutral formalin,Isolated abdominal skins kept in room temperature for 1～13 days were also observed for the purpose of studing the influence of the time elapsed after death on SRCA test results ABH substances were found in mucous membranes,mucous glands and prostate glands.ABH substances in those tissues were controlled by secrete status.ABH substances were also found in endothelia of blood vessels,stra- tified epithelia,acinar cells of pancreas and sweat glands.We firstly found that ABH substances were present in epithelia of pulmonary alveoli and epi- thelia of small bile duct in liver. Using SRCA test,the ABO blood type were correctly demonstrated in 11 isolated abdominal skins kept in room temperature for 1～13 days.
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Objective Study on the pattern of changes of TGF-?1 and type I receptor occurred in the experimental fluid percussion brain injury model for the purpose of providing the scientific basis for molecular pathological diagnosis, forensic identification, clinical treatment as well as further ascertaining the molecular mechanism of brain injury. Methods Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of injury groups were subjected to moderate lateral fluid percussion brain injury (0.2 MPa). The injury groups were then subdivided into 30min, 1h, 3h, 6h, 12h,1d, 3d and 7d sub-groups according to the time elapsed after injury. Immunohistochemistry SP method was used for studing the immunoreactivity of both TGF-?1 and T?R I factors. Results (1) In the brain of normal control and sham operation control groups, the low expression levels of TGF-?1 and T?R I were observed; (2) The gradual increase of TGF-?1 and T?R I immunoreactivity could be observed 1 to 3d after injury both in cortex and brain stem, and sustained at the high level up to 7d; (3) In hippocampus, the gradual increase occur during 12h to Id after brain injury, and sustained the high level at 3d, then declined at 7d. Conclusion The results suggested that brain injury induced the gene eypressions of the TGF-?1/ T?R I . The TGF-?1/ T?R I may contribute to maintance of nerve cell survival and the repair of damaged neural tissues after CNS injury and the patterns of their level change were quite regular and can be used for timing of injury in forensic medicine aspect.
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Two hybridoma cell lines (G8,E10) secreting monoclonal antibodies against human spermatozoa were established by fusing Sp2/0 mouse myeloma cells with splenocytes from Balb/c mice immunized with washed human spermatozoa. The results of the IIF, Elisa or peroxidase conjugated antibody im- munohistochemical staining demonstrated that both antibodies reacted with the spermatozoa in the human semen, testis, epididymus and vas deferens as well as the germ cells in the testis, cross reacted with the epithelial cells Of the thyroid gland, renal tubules, pancreatic ducts as well as the ilets of Langhans of the pancrease and not cross reacted with 33 normal human tissue cells, 9 different sources of body fluids and secretions as well as 8 different species of animal's spermatozoa. It is concluded that both monoclonal antibodies can be used for the species identification of semen stain in sex assault cases. The results of IIF demonstrated that each antibody bind to the localized region of the sperm cell surface. The antibody binding patterns were restricted to the acromosal cap, equatorial segment, postacromosal region, neck and the midpiece. The observed antibody binding patterns suggest that the human sperm surface is divided into a minimum of five domains. Both monoclonal antibodies were belong to IgG_1 subclass. The titre of Gs was 1∶512, while of E_(10) 1∶1024.
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Group-specific component(Gc), one of the polymorphic proteins of human plasma,has been isolated and purified from human plasma of Gc 2-2 phenotype by a procedure including DEAE-Sephadex-A50,Sephadex G100 and DEAE-Sephadex-A50 chromatography. The purified Gc identified by PAGE,SDS-PAGE and immunoelectrophoresis was homogeneous and reacted specifically with the commercial anti-Gc serum (DAKO) kit. The molecular weight of the purified Gc was about 58 kd.
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0.05).The exclusion probability of Gc is 34.8% and discrimination probability of Gc is 79.44%.There are not any significant difference of the distribution of Gc subtypes between the Hun population in Chengdu and those in Hong Kong and Japanese.The difference of the distribution of Gc subtypes between the Han population in Chengdu and those in Malaysia,Indonesia,India as well as the American caucasians,Belgians,Icelander and West German are sig- nificant. The phenotyping of Gc in 11 bloodstain samples kept in room temperature for twenty weeks were carried out successfully also using PAGIF followed by immunofixation method.