CN104087660B - Method for deducting postmortem interval of human body - Google Patents
Method for deducting postmortem interval of human body Download PDFInfo
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- CEAZRRDELHUEMR-UHFFFAOYSA-N gentamicin Chemical class O1C(C(C)NC)CCC(N)C1OC1C(O)C(OC2C(C(NC)C(C)(O)CO2)O)C(N)CC1N CEAZRRDELHUEMR-UHFFFAOYSA-N 0.000 description 2
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
- G01N15/0205—Investigating particle size or size distribution by optical means
- G01N15/0227—Investigating particle size or size distribution by optical means using imaging; using holography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
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Abstract
The invention discloses a method for deducting the postmortem interval of a human body, and belongs to the medical science examination field. The method comprises the following steps: extracting human gallbladder mucous membrane epithelial cells at different death times as samples, and smearing the samples; dyeing the smeared samples; carrying out image analysis on the dyed samples, wherein analysis parameters comprise index of density, gray scale parameters and geometric parameters of the nuclei of the gallbladder mucous membrane epithelial cells, and the gray scale parameters comprise average optical density, average gray scale and integral optical density; and carrying out statistics on the parameters to obtain linear regression equations of the parameters, and determining the postmortem interval according to one or more of the linear regression equations. The index of density, gray scale parameters and geometric parameters of the nuclei of the gallbladder mucous membrane epithelial cells are analyzed within 5-36h after human body death through method for deducting the postmortem interval of the human body to obtain a relation between each of the index of density, gray scale parameters and geometric parameters and PMI in order to deduct the postmortem interval of the human body.
Description
Technical field
The present invention relates to field of medical examination, more particularly to a kind of method for inferring human body postmortem interval.
Background technology
Human body postmortem interval (postmortem interval, PMI) infers the weight for being always forensic study
One of point, difficult point and focus, the especially deduction of early stage PMI, suspect, the dead medical treatment of arbitration are investigated in criminal investigation and are entangled
Confused grade have more in juridical practice realistic meaning.Objective, accurate early stage PMI deductions, can provide weight for Public Security Organss' criminal investigation
The objective clue wanted, is conducive to Public Security Organss to distinguish whether suspect has crime time, it is determined that or reduce scope of investigation, carry
Height investigation efficiency and accuracy.Additionally, the analysis and assessment inferred sometimes to natural disaster of postmortem interval, insurance and property
Inherit also most important.
DNA is the hereditary material with notable biochemical stability, is permanent with the somatic DNA content of species different tissues
Fixed.After body death, there is ischemia, anoxia in cell tissue, increase as oxygen-derived free radicals are generated, cell autodigestion, its
Structure is disintegrated, and DNA also gradually degrades, and the content of its DNA and nuclear metamorphosis also have regularity, show as
Core solidification, side collection, fragmentation, dissolving etc. change.Research worker is when postmortem interval is studied, and it is thin that its object of study is brain
Born of the same parents, splenocyte or hepatocyte, and research worker thinks that the DNA only in brain cell, splenocyte or hepatocyte can be accurately anti-
Reflect postmortem interval.
The content of the invention
Not for gallbladder reflection postmortem interval in order to solve the problems, such as prior art, the embodiment of the present invention is provided
A kind of method for inferring human body postmortem interval.The technical scheme is as follows：
The invention provides a kind of method for inferring human body postmortem interval, methods described includes：
The mucous membrane of gallbladder epithelial cell of people of different death times is extracted as sample, and by the sample smear；
The sample after by smear is dyeed；
The sample after to dyeing carries out graphical analyses, and the parameter of the analysis includes：The mucous membrane of gallbladder epithelium is thin
The nuclear special-shaped index of born of the same parents, grey parameter and geometric parameter, the grey parameter include average optical, average gray and
Integral optical density；
The parameter is counted, the equation of linear regression of the parameter is obtained, the equation of linear regression is：
YID=11.333XID- 33.338, wherein, YIDFor the postmortem interval inferred, XIDAbnormity for correspondence measurement refers to
Number；
YAG=0.679XAG- 54.277, wherein, YAGFor the postmortem interval inferred, XAGFor the average ash of correspondence measurement
Degree；
YAOD=-178.868XAOD+ 84.222, wherein, YAODFor the postmortem interval inferred, XAODIt is flat that correspondence is measured
Equal optical density；
YIOD=-6.422XIOD+ 91.398, wherein, YIODFor the postmortem interval inferred, XIODFor the product of correspondence measurement
Light splitting density；
One or more in the equation of linear regression determine postmortem interval.
Specifically, the mucous membrane of gallbladder epithelial cell of the sample clip from the people after death 5～36h.
Further, the method for the smear is：The sample for taking out is placed in into the gentamycin containing 400mg/L
In normal saline, first after death 5h is completed in the people time of sample described in clip, 0.5h is deposited in 18 DEG C of calorstats, so
Afterwards in moisture preservation box, clip 0.5cm × 1 part of sample described in 0.5cm × 1cm, and, by the sample of clip. every 1h clips once
Originally it is dipped in clear water and washs, takes out the sample and blotted with filter paper, the sample coating after blotting.
Further, by coating after the sample dyeed using inspiring confidence in your root dyeing method.
Specifically, your the root dyeing method that inspires confidence in includes：0.5h will be fixed in the smear immersion fixative of standby dye, taken
It is placed in after going out in 6N hydrochloric acid solutions and hydrolyzes 30min, be placed in after taking-up in distilled water and wash, then immerses the Schiff prepared by paramagenta
30min is placed in family name's reagent, with the sodium metabisulfite solution that concentration is 0.5% 2min is embathed, repetition is embathed three times, flowing water punching
After washing 5min, mounting is carried out.
Further, the fixative is Ka Nuoshi fixatives.
Specifically, described image analysis adopts color pathological image analysis system.
Further, the field of view of the smear under microscope high power objective after intake dyeing, with " bow " font
Shape as collection circuit, it is ensured that the image of collection takes at random 10 visuals field without repeating, without intersection per the smear,
Each visual field collection piece image.
Further, the geometric parameter includes area, average diameter, equivalent diameter and girth.
The beneficial effect that technical scheme provided in an embodiment of the present invention is brought is：The present invention provide deduction human body after death between
Every the method for time, by nuclear special-shaped index, grey parameter and the geometric parameter of analyzing mucous membrane of gallbladder epithelial cell, obtain
To special-shaped index, grey parameter and geometric parameter and PMI relations, so as to infer human body postmortem interval, additionally, being compared to
Other histiocytes, the nucleus of mucous membrane of gallbladder epithelial cell are easy to extract and preserve, and the process of sample is easier to be avoided
The impact of individual variation and extraneous factor, overcomes certain subjectivity, empirical, it is ensured that the accuracy of corrupt corpse,
Reliability and repeatability.
Description of the drawings
Technical scheme in order to be illustrated more clearly that the embodiment of the present invention, below will be to making needed for embodiment description
Accompanying drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the present invention, for
For those of ordinary skill in the art, on the premise of not paying creative work, can be obtaining other according to these accompanying drawings
Accompanying drawing.
Fig. 1 is the variation diagram of the nuclear area with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention；
Fig. 2 is the change of the nuclear average diameter with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Figure；
Fig. 3 is the change of the nuclear equivalent diameter with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Figure；
Fig. 4 is the variation diagram of nuclear all personal attendant PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention.
Fig. 5 is the change of the nuclear special-shaped index with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Figure
Fig. 6 is the change of the nuclear average gray with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Figure；
Fig. 7 is the change of the nuclear average optical with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Change figure；
Fig. 8 is the change of the nuclear integral optical density with PMI of mucous membrane of gallbladder epithelial cell provided in an embodiment of the present invention
Change figure.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing to embodiment party of the present invention
Formula is described in further detail.
Gallbladder is located at the pyriform pouch construction at liver rear under the rib of right, has concentration and stores the effect of bile.Gallbladder
Divide bottom, body, neck, pipe four, cervical region connects cystic duct.Gallbladder wall is by mucosa, muscle layer and adventitia up of three layers.The mucosa has typical case
Absorption-type cell feature, with stronger absorption and concentrating function, the mucosa also has muciparous function, in mucosa
In, except general extracellular, also endocrine cell.Inventor has found gallbladder this device during the present invention is realized
Official can also be used for inferring human body postmortem interval.Below using in conjunction with specific embodiments the present invention is described in detail.
Embodiment
Select Hubei province that there is the gallbladder tissue 30 of the in vitro people of definite death time for 2013, the dead's age is with 30
± 10 years old are in the majority, wherein, male 28 (accounts for the 93.33% of sum), and women 2 (accounts for the 6.67% of sum).Corpse before drawing materials
Body is stored in 16～25 DEG C of environment, and, at 14.5～25 DEG C, the time of drawing materials first is in human body for the experimental situation temperature drawn materials
After death 5h is completed.
Complete gallbladder is taken out under the conditions of aseptic operation, in being placed in the normal saline containing 400mg/L gentamycins,
0.5h is deposited in 18 DEG C of calorstats, then in the moisture preservation box of aseptic, clip 0.5cm × 1 part of 0.5cm × 1cm samples, and
Every 1h clips once, washing, cleaning blood plasma components and hemocyte will be swung in the sample immersion clear water of clip, takes out the sample
And blotted with filter paper, the sample after blotting is clamped with tweezers carries out smear, and is coated according to uniform straight line coating method.
The present invention in sample preparation procedure, without destruction mucous membrane of gallbladder epithelial cell morphosiss, without make gallbladder tissue
Unicellular homogenate suspension.
Smear is analyzed by special messenger's unified measurement, is respectively provided with identical system environmentss before collection every time, and image analyzer is each
After reopening, before measuring, it is intended to carry out yardstick calibration, optical density calibration.The step of all operations program, order one
Cause.
Wherein, gallbladder inner face is by mucosa, muscle layer and adventitia up of three layers, and constituent is thin except mucomembranous cell and endocrine
It is extracellular, also including kytoplasm mitochondrial, rough endoplasmic reticulum, MG, blood vessel, lymphatic vessel and nerve etc., gallbladder collection and
In sample fritter cutting procedure, all can be different degrees of by antibacterial or microorganism pollution, some compositions inherently carry micro-
Biological or antibacterial, so an any of which part is contaminated, can all affect the other compositions of surrounding, the present invention to take out complete
Gallbladder is placed in the normal saline of 400mg/L gentamycins, and antibacterial for dispelling sample surface that so can be more thoroughly etc. may
The material of interference experiment result, while and cell interior structure will not be destroyed, do not affect cell extraction and smear effect.
Mucous membrane of gallbladder epithelial cell kytoplasm acidophilia, and water and inorganic salt (mainly Na+) in bile are absorbed, it is placed at
0.5h is deposited in 18 DEG C of calorstats, the balance of cell acid-base value and electrolyte is on the one hand can ensure that, makes cell that acid will not occur
Alkali is unbalance；On the other hand the freshness of sample is can ensure that, cell surface and internal moistening is made, more serious dehydration is not occurred and is showed
As farthest ensureing the integrity of sample cell.
According to aforesaid operations method, sample is sterilized more thoroughly, and smoothly, cellularity is complete for cell extraction process, extracts effect
Fruit is good, and smear effect reaches the requirement of experiment setting, it is ensured that the objectivity of experimental result, scientific and accuracy.
After sample smear, using the cell for inspiring confidence in your root (Feulgen-Vans) colouring method display mucous membrane of gallbladder epithelial cell
Core, for the quantitative measurement of nucleus DNA.
Using inspiring confidence in comprising the following steps that for your root dyeing method：
Smear immersion Ka Nuoshi (Carnoy) fixative of standby dye is fixed into 0.5h, water in 6N hydrochloric acid solutions is placed in after taking-up
Solution 30min, is placed in distilled water after taking-up and washs, then immerses placement in Xi Fushi (Schiff) reagent prepared by paramagenta
30min, with the sodium metabisulfite solution that concentration is 0.5% 2min is embathed, and repetition is embathed three times, and flowing water is rinsed after 5min, is used
Conventional neutral glue (such as resinenes) carries out mounting, all smears of each gallbladder tissue in after death 5～36h, is all from
Same individuality, must disposably concentrate and complete dyeing.
Mucous membrane of gallbladder epithelial cell smear time earliest 3～5h after death first, therefore, the significant figure that the present invention is selected
According to unified from 5h, shown by the early stage animal experiment study of the present invention, DNA degradation is in almost plateau phenomenon in 0～6h.Together
When, because under the experimental situation of the present invention, the self-dissolving of dead 36h descendants gallbladder tissue is seriously more difficult by thin smear, therefore grinds
Study carefully selection of time to complete in after death 36h.
Wherein, the preparation of Carnoy fixatives is anhydrous alcohol according to volume ratio：Chloroform：Glacial acetic acid=6:3:1.
Using the CMIAS color pathological image analysis systems of Motic Med 6.0, (Beijing Mike Audi image technique is limited
Company provides), the field of view of the smear under microscope high power objective (400 ×) after intake dyeing, with the shape of " bow " font
Shape is used as collection circuit, it is ensured that without repetition, without intersection, every smear takes at random 10 visuals field to the image of collection, and each visual field is adopted
Collection piece image.
Specifically, ensure compliance with it is statistical under the premise of, it is desirable to increase sample size as far as possible.Provide in coincidence statistics metering
On the premise of material significance test result, the present invention at least needs 11 individual specimen data to react overall situation, together
When need by every smear be divided into 9 visuals field and take at random 6 visuals field, per two field picture (adopting a two field picture at random in each visual field) with
Machine takes 10 cells, you can meet the needs to overall inference.
The parameter of analysis includes：Mucous membrane of gallbladder epithelial cell nuclear special-shaped index (Index of density,
ID), grey parameter and geometric parameter, wherein, grey parameter includes：Average optical (Average Optical Density,
AOD), average gray (Average Gray, AG) and integral optical density (Integral Optical Density, IOD)；Geometry
Parameter includes：Area (Area, A), average diameter (Average Diameter, AD), girth (Perimeter, P) and equivalent straight
Footpath (Equivalent Diameter, ED)
Above-mentioned parameter is measured, then respectively selective body carries out automatic measurement depending on analysis system and DNA analysis system, obtains
Obtain corresponding related data.
Data carry out dependency and linear regression analyses using SPSS10.0 statistical softwares, specific as follows：
The nuclear geometric parameter of mucous membrane of gallbladder epithelial cell
Each final geometric parameter values for counting are 30 all of sample in the time point of same PMI and are detected
The average of a certain parameter of cell.
The average of each geometric parameter measurement of nucleus of the difference PMI mucous membrane of gallbladder epithelial cells of table 1
Experimental result：From the parameter in table 1, geometric parameter is with postmortem interval without correlation statistically.
Geometric parameter is counted, geometric parameter is obtained with postmortem interval without statistical dependency.
The nuclear area of mucous membrane of gallbladder epithelial cell, in human body, after death 5～15h is in obvious ascendant trend, in 18～30h
In a platform fluctuation, it is on a declining curve in 30～36h, referring to Fig. 1, and in after death 5～36h its change with corresponding PMI without
Correlation statistically.
Normal size (6 of the measured value of the nuclear average diameter of mucous membrane of gallbladder epithelial cell in mature lymphocyte
～8 μm) scope fluctuates up and down, and referring to Fig. 2, it is in after death 5～36h with corresponding PMI without correlation statistically.
The nuclear equivalent diameter of mucous membrane of gallbladder epithelial cell, human body after death 5～15h be in obvious ascendant trend, 18
～30h is in a platform, and 30～36h is on a declining curve, and referring to Fig. 3, its equivalent diameter overall variation trend is similar with area parameters,
It changes with corresponding PMI without correlation statistically in after death 5～36h.
The nuclear girth of mucous membrane of gallbladder epithelial cell, with corresponding PMI without correlation statistically in after death 5～36h,
The average of its girth fluctuates about 29 μm, referring to Fig. 4.
The nuclear special-shaped index of mucous membrane of gallbladder epithelial cell
After death in 5～36h, the nuclear special-shaped index (ID) of the mucous membrane of gallbladder epithelial cell of human body is with PMI's for human body
Extend and it is in rising trend, referring to Fig. 5.The corresponding correlation coefficient square of concrete statistics, correct correlation coefficient square, standard error,
Equation of linear regression and its fit conformity test the results are shown in Table 2 and table 3.As a result show：Have very strong between special-shaped index and PMI
Correlation statistically, and there is linear relationship.
The correlation analysiss of the special-shaped index of table 2 and PMI
The linear regression analyses of the special-shaped index of table 3 and PMI
From table 2 and table 3, there are very strong correlation statistically, and R between special-shaped index and PMI2For 0.967, R2For
The linear relationship of coefficient of determination, special-shaped index and PMI, the statistically significant (p of classical prescription difference inspection<0.01, p value is credible result
One declining indicator of degree), show that the special-shaped exponent pair PMI has Regress estimate, i.e., the special-shaped index exists clear and definite with PMI
Quantity dependence.Wherein, equation of linear regression is YID=11.333XID-33.338(YIDFor the PMI for inferring；XIDFor correspondence
The ID averages of PMI measurements), the partial regression coefficient in its regression equation, Jing statistics null hypothesises inspection (T test) show：
Refuse null hypothesises (P in the level of ɑ=0.01<0.001), illustrate there is linear relationship between special-shaped index and PMI.
The nuclear grey parameter of mucous membrane of gallbladder epithelial cell
Each final grey parameter value for counting is 30 all of sample in the time point of same PMI and is detected
The average of a certain parameter of cell.
The each grey parameter of nucleus of the difference PMI mucous membrane of gallbladder epithelial cells of table 4 measures average
The CMIAS color pathological image analysis systems of Motic Med 6.0 are only required and are accurate to one decimal place number, make
Obtain AG values and closely retain one decimal place.
After death in 5～36h, the nuclear average gray of mucous membrane of gallbladder epithelial cell is in rising to human body with PMI prolongations
Trend, referring to Fig. 6.The corresponding correlation coefficient square of statistics, correction correlation coefficient square, standard error, equation of linear regression and its
Fit conformity test the results are shown in Table 5 and table 6.
The correlation analysiss of the average gray of table 5 and PMI
The linear regression analyses of the average gray of table 6 and PMI
From table 5 and table 6, there is very strong correlation statistically (R between average gray and PMI2For 0.921), averagely
The linear relationship of gray scale and PMI, the statistically significant (p of classical prescription difference inspection<0.01), show that the average gray has back to PMI
Return contribution, i.e., the parameter has clear and definite quantity dependence with PMI.The equation of linear regression for drawing is：YAG=0.679XAG–
54.277(YAGFor the PMI for inferring；XAGFor the AG averages of average gray correspondence PMI measurements), the partial regression system of the regression equation
Number, Jing statistics null hypothesises inspection (T test) show：Refuse null hypothesises (P in the level of ɑ=0.01<0.001), explanation
There is linear relationship between average gray and PMI.
Human body is after death in 5～36h, is in that decline becomes extending with PMI for the average optical of mucous membrane of gallbladder epithelial nucleus
Gesture, referring to Fig. 7, the corresponding correlation coefficient square of statistics, correction correlation coefficient square, standard error, equation of linear regression and its matches somebody with somebody
Appropriate degree assay is shown in Table 7, table 8.
The correlation analysiss of the average optical of table 7 and PMI
The linear regression analyses of the average optical of table 8 and PMI
From table 7 and table 8, there is very strong correlation statistically (R between average optical and PMI2For 0.921).It is flat
The linear relationship of equal optical density and PMI, the statistically significant (p of classical prescription difference inspection<0.01), show that the parameter has back to PMI
Return contribution, i.e., the parameter has clear and definite quantity dependence with PMI.The equation of linear regression for drawing is：YAOD=-
178.868XAOD+84.222(YAODFor the PMI for inferring；XAODFor the average optical average of average optical correspondence PMI measurements),
Partial regression coefficient in its regression equation, Jing statistics null hypothesises inspection (T test) show：Refuse in the level of ɑ=0.01
Null hypothesises (P<0.001), illustrate there is linear relationship between average optical and PMI.
After death in 5～36h, the integral optical density of mucous membrane of gallbladder epithelial nucleus is in that decline becomes with the prolongation of PMI to human body
Gesture, referring to Fig. 8, the corresponding correlation coefficient square of concrete statistics, correct correlation coefficient square, standard error, equation of linear regression and
Its fit conformity test the results are shown in Table 9, table 10.
The correlation analysiss of the integral optical density of table 9 and PMI
The linear regression analyses of the integral optical density of table 10 and PMI
From table 9 and table 10, there is very strong correlation statistically (R between integral optical density and PMI2For 0.892).Product
The linear relationship of light splitting density and PMI, the statistically significant (p of classical prescription difference inspection<0.001), show that the parameter has back to PMI
Return contribution, i.e., the integral optical density has clear and definite quantity dependence with PMI.Equation of linear regression is YIOD=-6.422XIOD
+91.398(YIODFor the PMI for inferring；XIODFor the IOD averages of integral optical density correspondence PMI measurements), its regression equation is biased back to
Coefficient Jing statistics null hypothesises inspection (T test) are returned to show：Refuse null hypothesises (P in the level of ɑ=0.01<0.001),
Illustrate there is linear relationship between integral optical density and PMI.
One or more in equation of linear regression determine postmortem interval, specifically, by linear regression
When equation obtains the postmortem interval inferred, an equation of linear regression can be adopted, it would however also be possible to employ multiple linear regressions
Equation, if the postmortem interval of multiple deductions of gained is inconsistent, take multiple deductions postmortem interval it is average
Value, the meansigma methodss are the postmortem interval inferred.
The method of the deduction human body postmortem interval that the present invention is provided, in terms of the selection of the species of sample, the present invention
With the mucous membrane of gallbladder epithelial cell of people as first-selected object of study, gallbladder composition cell is mainly mucomembranous epithelial cell, tissue-derived
It is relatively single, its corresponding iuntercellular difference and disturb less, and for other internal organs, its pathology pathological changes is few.Together
When the present invention early stage zoopery also confirm that gallbladder is one of best selected organ of graphical analyses.Human body after death 5～
In 36h, by nuclear special-shaped index, grey parameter and the geometric parameter of analyzing mucous membrane of gallbladder epithelial cell, abnormity is obtained
Index, grey parameter and geometric parameter and PMI relations, so as to infer human body postmortem interval, additionally, being compared to other groups
Cell is knitted, the nucleus of mucous membrane of gallbladder epithelial cell are easy to extract and preserve, the process of sample is easier to avoid individual diversity
Different and extraneous factor impact, overcomes certain subjectivity, empirical, it is ensured that the accuracy of corrupt corpse, reliability
And repeatability.
The foregoing is only presently preferred embodiments of the present invention, not to limit the present invention, all spirit in the present invention and
Within principle, any modification, equivalent substitution and improvements made etc. should be included within the scope of the present invention.
Claims (1)
1. it is a kind of infer human body postmortem interval method, it is characterised in that methods described includes：
The mucous membrane of gallbladder epithelial cell of people of different death times is extracted as sample, and by the sample smear, the sample
Clip is from the mucous membrane of gallbladder epithelial cell of the people after death 5～36h, the method for the smear：In aseptic operation condition
It is lower to take out complete gallbladder, in being placed in the normal saline of the gentamycin containing 400mg/L, deposit in 18 DEG C of calorstats
0.5h, then in moisture preservation box, clip 0.5cm × 1 part of sample described in 0.5cm × 1cm, first the time of sample described in clip exist
After death 5h is completed the people, and, the sample of clip is dipped in clear water and is washed every 1h clips once, takes out the sample
And blotted with filter paper, the sample coating after blotting, and be coated according to uniform straight line coating method；
The sample after by smear is dyeed using your root dyeing method is inspired confidence in, and your the root dyeing method that inspires confidence in includes：Will be standby
0.5h is fixed in the smear immersion Ka Nuoshi fixatives of dye, is placed in after taking-up in 6N hydrochloric acid solutions and is hydrolyzed 30min, after taking-up
Be placed in distilled water wash, then immerse by paramagenta prepare Xi Fushi reagents in place 30min, with concentration be 0.5% it is inclined
Sodium bisulfite solution embathes 2min, and repetition is embathed three times, and flowing water is rinsed after 5min, and with conventional neutral glue mounting is carried out, each
All smears of the gallbladder tissue in after death 5～36h, are all from same individuality, must disposably concentrate and complete dyeing, wherein blocking
The preparation of Nuo Shi fixatives is anhydrous alcohol according to volume ratio：Chloroform：Glacial acetic acid=6:3:1；
The sample after to dyeing carries out graphical analyses, the employing color pathological figure using color pathological image analysis system
As the method that analysis system carries out graphical analyses is：The image of the smear under microscope high power objective after intake dyeing is regarded
Open country, using the shape of " bow " font as collection circuit, it is ensured that the image of collection without repeating, without intersection, and per the smear with
Machine takes 10 visuals field, and each visual field collection piece image, the parameter of the analysis includes：The mucous membrane of gallbladder epithelial cell it is thin
The special-shaped index of karyon, grey parameter, the grey parameter includes average optical, average gray and integral optical density；
The special-shaped index, average optical, average gray and integral optical density are counted respectively, the abnormity is obtained and is referred to
The equation of linear regression of number, average optical, average gray and integral optical density, the equation of linear regression is：
YID=11.333XID- 33.338, wherein, YIDFor the postmortem interval inferred, XIDFor the special-shaped index of correspondence measurement；
YAG=0.679XAG- 54.277, wherein, YAGFor the postmortem interval inferred, XAGFor the average gray of correspondence measurement；
YAOD=-178.868XAOD+ 84.222, wherein, YAODFor the postmortem interval inferred, XAODThe average light of correspondence measurement
Density；
YIOD=-6.422XIOD+ 91.398, wherein, YIODFor the postmortem interval inferred, XIODFor the integral light of correspondence measurement
Density；
One or more in the equation of linear regression determine postmortem interval.
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Non-Patent Citations (2)
| Title |
|---|
| 人脾细胞核几何参数变化与早期死亡时间的相关性研究;舒细记等;《法医学杂志》;20041231;第20卷(第3期);140-142 * |
| 比较人和兔细胞DNA降解来推断人死亡时间的可行性研究;罗国厂;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20080115(第1期);E076-7 * |
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