Objective To study the relationship between the ultrastructural changes of corneal matrix collagen fibrils diameter and the postmortem interval.Methods Twenty-eight rabbits were killed by the method of air embolism and the samples of cornea were separately cut down in 0,6,12,18,24,48 and 72h after death.The ultrastructure of the collagen diameter was observed by transmission electro microscopy(TEM)and the 4 parameters of area(Y1),perimeter(Y2),average-diameter(Y3)and mean-diameter(Y4)were measured by a computer image technique and analyzed by Excel 2000 and SPSS 10.0 software.Results The 4 parameters of Y1,Y2,Y3 and Y4 were respectively increased from 1 131?53nm to 1 628?26nm,from 132.8?23nm to167.5nm,from 38nm to 45nm and from 37.71?6nm to 44.89?5nm within 72 hours after the death.Conclusion The corneal matrix collagen diameter increase regularly with the postmortem interval in 72h after death,and the parameters of Y1,Y2,Y3 and Y4 could be used as a new reference mark for timing the postmortem interval after death.

ObjectiveTo study the effect and clinical meaning and pathogenic of extinction on cerebral cortex lesion. MethodsTactile extinction to classical extinction test(simple stimuli)and modified Quality extinction test(QET)were used. ResultsThe positive rate of ex-tinction using modified QET was higher than that of simple stimuli; left cortex lesion could result in extinction both in the contralateral side andthe ipsilateral side;left frontal sites lesion caused ipsilateral extinction. Conclusion: The modified QET was more sensitive than simplestimuli; modified QET showed extinction has competiting and restricting effect at 2 different levels, the mechanism of extinction is correlatedto competition and adventage restraint of diplo-hand cerebral hemisphere and liaise correliation fiber of diplo-hand cerebral hemisphere; thehighesttthe positive rate of extinction was observed in left parietal lesions.

Objective To observe and measure the facial nerve canal(FNC) in curved planar reformation by multislice spiral CT.Methods High resolution computed tomography (HRCT)of temporal bone was performed in 40 cases (80 ears) by multislice spiral CT. Curved planar reformation of FNC was performed separately in axial, coronal and sagittal plane of multiplanar reconstruction. The morphology and measurement of FNC were studied.Results The mean length of FNC was (29.73?1.07)mm. The mean length of the labyrinthine, tympanic and mastoid segment was (5.6?0.74)mm, (10.66?0.79)mm and (13.47?1.01)mm respectively. The mean diameter of the l-abyrinthine, geniculate fossa, tympanic and mastoid segment was (0.76?0.16)mm, (2.37?0.63)mm, (1.03?0.16)mm and (1.57?0.31)mm respectively. The mean angle of first and second genu was 67.04??9.41? and 114.25??8.44? respectively. The distance between the tympanic segment and ampulla of the lateral semicircular canal was (0.57?0.19)mm. The distance between the tympanic segment and arch of the lateral semicircular canal was (1.59?0.61)mm. The mean angle between the tympanic segment of the facial nerve canal and the lateral semicircular canal was 10.63??3.60?. The tympanic course of the facial nerve canal formed an angle of 34.65??5.39? with the sagittal plane. Dehiscences was found in 89% FNC, 73.7% of them only located in the tympanic portion and 26.3% both in the mastoid and tympanic portions. Conclusion Curved planar reformation of FNC by multislice spiral CT is the good method to study the anatomy of FNC,and in combination with HRCT axial scan and multiplanar reconstrution can obtain more valuable information.

AIM : To investigate the effects of Ca 2+ /CaM dependent calcineurin(CaN) signaling pathway on cardiomyocytes hypertrophy of rat induced by neuropeptide Y(NPY). METHODS : Cardiomyocytes of neonatal Wistar rats were cultured with NPY of various concentrations (10,100 nmol?L -1 ). Cyclosporine A (CsA) was used to inhibit the activity of CaN. The methods of 3H Leu incorporation was used to assess protein synthesis rate in cardiomyocytes. Western blot and histochemistry were used to measure CaN protein expression and CaN activity in cardiomyocytes. RESULTS : 3 H Leu incorporation of cardiomyocytes were increased significantly by 100 nmol?L -1 NPY ( P

Objective To investigate the protective effect of methylene blue (MB) on blood-brain barrier (BBB) injury after focal cere-bral ischemia-reperfusion in rats. Methods 18 male Sprague-Dawley rats were randomly divided into sham-operated group (n=6), model group (n=6) and MB treatment group (n=6). The left middle cerebral arteries were occluded for 1 hour and reperfused. MB was infused intra-venously immediately after reperfusion (3 mg/kg) and again 2 hours post-reperfusion (1.5 mg/kg), while normal saline was administered in the model group. The sham-operated group was treated as same as the model group without occlusion and infusion. HE staining was used to observe the histological injury in the cortex around the infarcted region 47 hours after reperfusion, while albumin immunohistochemistry was used to evaluate the permeability of the BBB, and immunohistochemistry and double immunofluorescence staining were used to exam-ine the expressions of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4). Results HE staining showed that cells and blood ves-sels were not intact in the cortex around the infarcted region in the model group and they were better in the MB treatment group. The expres-sions of the albumin, GFAP and AQP-4 were higher in the model group than in the sham-operated group (P<0.01), and were lower in MB treatment group than in the model group (P<0.05). The double immunofluorescence staining showed the colocalization of GFAP and AQP-4 in the astrocytes. Conclusion MB may ameliorate the BBB disruption induced by focal cerebral ischemia-reperfusion through reducing glio-cyte proliferation and down-regulation of AQP-4 expression in rats.

As a rising histological specimen carrier,digital slide has advantages of easy search and fast browse.By digital scanning and stitching of traditional slides and uploading traditional slides as well as pathological information to network servers,the construction of digital slides data can take its own superiority to assist forensic medicine teaching.Combined with our research and teaching experience,this article discussed the application prospects of digital slide technology and digital slides data so as to provide references for the improvement and reformation of forensic medicine education.

Objective To do research on Multi-points EGFR gene mutation and heterogeneity in lung adenocarcinoma and its influence on the prognosis,to analyze EGFR gene mutation and its heterogeneity influence on patients'overall prognosis.Methods The clinical features of patients with lung adenocarcinoma at stage Ⅲa from January 2006 to January 2007 at our institution were retrospectively reviewed.The primary lung tumors and corresponding metastatic lymph nodes tissue specimens were obtained by surgery.The adenocarcinoma primary nodes and corresponding metastatic lymph nodes EGFR mutation were detected by amplification refractory mutation system (ARMS).Univariate analysis and multivariate analysis by Cox proportional-hazard model were used to analyze the impact of EGFR mutation and its heterogeneity as influential factor on patients 'prognosis.Results 76 patients with the adenocarcinoma primary nodes and corresponding metastatic lymph nodes were detected by epidermal growth factor receptor (EGFR) mutation.40 patients with EGFR mutation were detected (40/76,52.63％).There were 9 specimens out of 40 who had lung primary nodes and corresponding metastatic lymph nodes EGFR gene heterogeneity (9/40,22.5％).Log-Rank univariate analysis showed that there was no significant difference in overall survival period between EGFR mutation patients and wild-type patients(x2 =0.382,P =0.537),but there was significant difference in illnessfree progression period(x2 =4.147,P =0.042).Gene heterogeneity factor does not affect on the overall survival period and illness-free progression period of the patients with EGFR gene mutation (x2 =1.774,P =0.183 ;x2 =1.249,P =0.264).Multivariate analysis by Cox proportional-hazard model showed that EGFR gene mutation is not the independent risk factor that has 赵 impact on the prognosis of patients with lung adenocarcinoma.Conclusion Assessment of EGFR gene mutations in a single-point specimen can not reflect the whole EGFR gene mutation status,which may probably cause difference between targeted drugs'predicted effect and its actual usage effect.

BACKGROUND:Recombinant fusion protein is cascaded by staphylokinase and hirudin according to the thrombin cognition sequence, and has double functions and a molecular weight of 23 ku. The recombinant fusion protein can be highly expressed in the engineering bacteria at high-density fermentation. OBJECTIVE:To construct and purify recombinant staphylokinase-hirudin fusion protein in the engineering bacteria after high-density fermentation, and to explore the feasibility of construction and the expression value. METHODS:The engineering bacteria were cultured at high density and staphylokinase-hirudin fusion protein was induced to express. The bacteria were centrifuged and ultrafiltrated after repeated freezing and thawing. The supernatant was colected with ion exchange chromatography method. The staphylokinase-hirudin fusion protein was isolated and purified, then the fibrinolytic activity and expression in bacteria were observed. RESULTS AND CONCLUSION: The engineering bacteria were cultured and the fusion protein was induced at 17 hours. The results showed that, staphylokinase-hirudin fusion protein expression was detected at 0.5 hours after induction, and the expression levels were increased as the fermentation time; at 20 hours, the expression level reached the peak. The dried weight of the bacteria was 32.20 g/L and the expression level of target proteins was 1.48 g/L. After purification, the purity of recombinant staphylokinase-hirudin fusion protein was as high as 98%, fibrinolytic activity was about 2.6×104 IU/mg, the probability of activity recovery was 56%. The purification process of recombinant staphylokinase-hirudin fusion protein is convenient, less time, repeatable and alows large-scale production.

Objective To screen an ssDNA aptamer for rabbit mesenchymal stem cells (MSCs) and to identify the ability of the aptamer to recognize MSCs of a variety of species origin.Methods MSCs were isolated from the thigh bone of immature rabbits and identified by induced osteogenic and adipogenic differentiation,respectively.Aptamers were screened by cell SELEX (systematic evolution of ligands by exponential enrichment) technique targeting to isolated MSCs.Enrichment of the 5th pool was evaluated through binding assay of FAM modified pool to MSCs by confocal microscopy.The enriched 5th pool was then cloned into pGE-T vector and the cloned sequences were determined randomly.The candidates were chosen based on primary sequence conservation and predicted secondary structure by RNA structure and MEME online analysis.Flow cytometry analysis was used to identify the aptamers binding to MSCs of rabbit, rat, and human origin.Results The isolated MSCs had the potential of osteogenic differentiation and adipogenic differentiation under certain conditions.Aptamer 5-1-12 from 5th enriched pool was characterized as MSCs recognizing aptamer binding to MSCs of rabbit, rat and human origin.Conclusion Aptamer 5-1-12 that recognizes MSCs of different species origin is obtained through live cell-SELEX.

Objective To explore the expression of aquaporin 4 (AQP4) in primary and secondary cultured rat astrocytes. Methods Rat cortical astrocytes from a newborn (one day) Wistar rat were cultured. Astrocytes were identified with immunofluorescence staining of glial fibrilillary acidic protein (GFAP). The expression of AQP4 was determined with real-time quantitative polymerase chain reaction and immu-nofluorescence staining three, five, seven and nine days of primary culture, and nine days of secondary culture. Results The purity of GFAP-positive cells was more than 95％. The expression of AQP4 mRNA was found three days of primary culture, remained unchanged five days of primary culture (P>0.05), and increased seven and nine days of primary culture (P<0.05). The expression of AQP4 mRNA was not different between nine days of primary culture and nine days of secondary culture (P>0.05). AQP4 immunofluorescence staining showed the same trend of AQP4 mRNA. Conclusion AQP4 may express since three days of primary culture in rat astrocytes in vitro, and increase slowly until nine days of primary culture.