Function of IL-33 in Wound Age Estimation of Skin Wounds in Mice / 法医学杂志

Objective To explore the application value of interleukin-33 （IL-33） in wound age estimation in forensic practice by observing the sequential changes of IL-33 after skin wound. Methods Skin wound models were generated on the back of mice with a round file of 5 mm in diameter. Skin samples of the same size were taken from the same parts of mice in control group and injury group 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d and 10 d after skin wound. Hematoxylin-eosin （HE） staining method was applied to observe the morphological changes in the recovering process after skin wound. Western blotting, immunohistochemistry staining and double immunofluorescence staining methods were applied to detect the expression changes of IL-33 in the skin wound samples. Results The results of Western blotting showed that the expression of IL-33 protein decreased slightly at 3 h after skin wound, increased gradually at 6 h after skin wound, and reached the peak value at 3 d, then decreased gradually. Immunohistochemistry staining results showed that faint positive expression of IL-33 was observed in epidermis, hair follicles, sebaceous glands and dermal resident cells of the control group skin. The positive cell rate of IL-33 increased at 3 h after skin wound and reached the peak value at 3 d, then decreased gradually. The results of double immunofluorescence staining showed that the majority of IL-33 positive cells from 1 d to 3 d after wound were macrophages, while the majority of IL-33 positive cells from 5 d to 7 d after wound were myofibroblasts. In addition, the results of HE staining showed that the wound healing process of the skin wound model was consistent with the pathological development law of inflammation. Conclusion IL-33 could become a reference index for wound age estimation of skin wound in forensic practice.

Application of Virtual Anatomy Technology in Postmortem Examination of Medical Dispute Cases / 法医学杂志

Objective To discuss the application value of CT scanning technology in cause of death determination of medical dispute cases. Methods From July 2017 to December 2018, postmortem CT imaging data of 12 medical dispute cases were collected. CT imaging diagnosis results and anatomy findings as well as differences between antemortem and postmortem CT diagnosis were compared. The advantages and disadvantages of CT routine tests of the cadavers in terms of the diagnosis of disease and damage were analyzed. Results The comparison between CT imaging diagnosis and anatomical findings showed that CT scans had advantages in the diagnosis of disease and damage with large differences in density changes, such as atelectasis, pneumonia, calcification, fracture and hemorrhage, etc. The comparison of CT diagnosis in antemortem and postmortem examination showed that the cadavers of medical dispute cases were well preserved and that postmortem CT scan was meaningful for the diagnosis of antemortem diseases. Conclusion Virtual anatomy technology has a relatively high application value in postmortem examination of medical dispute cases. It can provide effective information for the appraisers before the autopsy and can also provide a reference for cause of death analysis when the anatomy cannot be performed.

Relationship between the Number of Neutrophils and Myofibroblasts during Diabetic Wound Healing and Wound Age / 法医学杂志

Objective To investigate the sequential changes of the number of neutrophils and myofibroblasts during diabetic wound healing, and discuss its application value in wound age estimation. Methods Diabetic DB mice and mice of the same age in the normal control group were selected, a wound healing model was established, wound samples were taken at different time points, while the number of neutrophils and myofibroblasts during diabetic wound healing were determined by immunohistochemical staining technique. Results The number of infiltrated neutrophils in the wounds of control and diabetic groups reached the peak respectively at 12 h and 5 d after injury. Compared with the control group, the number of neutrophils in the diabetic group decreased significantly from 6 h to 1 d after injury, but increased markedly from 5 d to 14 d. From 5 d to 10 d after injury, the average number of neutrophils at high magnification in wounds of the diabetic group was over 30, while that of neutrophils in wounds of the control group was less than 20. Myofibroblasts appeared in wounds from 3 d to 14 d after injury in the control group and from 5 d to 14 d after injury in the diabetic group. The difference in the number of myofibroblasts in wounds between control group and diabetic group from 3 to 7 d after injury had statistical significance. Conclusion In comparison with normal wound healing, the number of neutrophils and myofibroblasts during diabetic wound healing shows different sequential changes. The results of this study can provide reference for wound age estimation of patients with severe diabetes.

Time-dependent Expression and Distribution of AChE during the Skin Incised Wound Healing in Mice / 法医学杂志

Objective To study the time-dependent expression and distribution of acetylcholinesterase （AChE） during skin incised wound healing in mice, and discuss its effect in wound healing as well as the feasibility of using it as a reference index for wound age estimation. Methods A total of 45 C57BL/KsJ mice were randomly divided into one control group and eight incised groups. The skin incised wound model was established in the incised groups with samples of skin wounds taken at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d post-injury respectively, while the uninjured skin tissue was extracted in the control group. Expression and distribution of AChE in skin samples were detected by immunohistochemistry, double immunofluorescence and Western blotting. Results Immunohistochemistry results indicated that AChE was mainly detected in infiltrating polymorphonuclear cells （PMNs） 6 to 12 h post-injury. A large number of AChE-positive mononuclear cells （MNCs） were observed 1 to 3 d post-injury. The AChE-positive cells were mainly fibroblastic cells （FBCs） 5 to 14 d post-injury. The ratio of the AChE-positive cells increased initially 6 h post-injury, and reached the peak at 1 d post-injury. Double immunofluorescent staining showed that the majority of AChE-positive MNCs and FBCs expressed macrophage marker and myofibroblast marker, respectively. Western blotting results showed that the relative expression level of AChE in the incised group was higher than that in the control group averagely, reached the peak at 1 d post-injury, then reached a second peak at 7 d post-injury. Conclusion The expression of AChE is found in PMNs, macrophages and myofibroblast during skin wound healing, which indicates it might be involved in the adjustment of inflammatory response and fibrotic repair after injury. Moreover, combined use of various methods for the detection of the expression of AChE would provide reference for skin wound age estimation.

Expression of Annexin A1 during Skin Incised Wound Healing in Mice / 法医学杂志

OBJECTIVES@#To investigate the expression changes of annexin A1 （ANXA1） during the process of skin incision healing, and to explore its expression and function during skin injury repair.@*METHODS@#The skin injury model of mice was prepared, and skin tissues of the controls and the injured group at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d after injuries were taken. The morphological changes of the wound were observed by hematoxylin-eosin （HE） staining, and the expression of ANXA1 was detected by immunohistochemistry （IHC） and Western blotting.@*RESULTS@#HE staining showed normal healing of skin wounds. IHC results revealed that ANXA1 was expressed in the epidermis, hair follicle, sebaceous gland and vascular endothelium. In the injured group, the expression of ANXA1 was enhanced in epidermis and skin appendages around the wound 6-12 h after injury, and ANXA1 was also highly expressed in neutrophils and a small number of mononuclear cells. ANXA1 was mainly positively expressed in monocytes, neovascular endothelial cells and fibroblasts, and small amount of fibroblasts at 1-3 d, 5-10 d, and 14 d after injury, respectively. Western blotting showed that, compared with the controls, the expression of ANXA1 was significantly increased at 6 h after injury, peaked at 1 d, and then decreased gradually in the injured group.@*CONCLUSIONS@#ANXA1 may be involved in the regulation of skin damage repair, with time-dependent expression during skin wound healing, and thus is expected to be a biological marker for inferring the wound formation time.

Relationship between FoxO1 Expression and Wound Age during Skin Incised Wound Healing / 法医学杂志

Objective To investigate FoxO1 expression and its time-dependent changes during the skin incised wound healing. Methods After the establishment of the skin incised wound model in mice, the FoxO1 expression of skin in different time periods was detected by immunohistochemistry and Western blotting. Results Immunohistochemistry staining showed that FoxO1 was weakly expressed in a few fibroblasts of epidermis, hair follicles, sebaceous glands, vessel endothelium and dermis in the control group. The FoxO1 expression was enhanced in the epidermis and skin appendages around the wound during 6-12 h after injury, which could be detected in the infiltrating neutrophils and a small number of monocytes. FoxO1 was mainly expressed in monocytes during 1-3 d after injury, and in neovascular endothelial cells and fibroblasts during 5-10 d. On the 14th day after injury, the FoxO1 expression still could be detected in a few fibroblasts. The Western blotting results showed that the FoxO1 expression quantity of the tissue samples in injury group was higher than in control group. The FoxO1 expression peaked at 12 h and 7 d after injury. Conclusion FoxO1 is time-dependently expressed in skin wound healing, which can be a useful marker for wound age determination.

Study on the mechanism of how curcumin improves pulmonary vascular remodeling associated with chronic pulmonary arterial hypertension / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of how curcumin improves pulmonary vascular remodeling associated with chronic pulmonary arterial hypertension.</p><p><b>METHODS</b>The model of chromic hypoxia hypercapniapulmoary remodeling was made. Twenty-four male rats were randomly divided into 4 groups (n = 6): group I (normoxia control group), group II (hypxia and hypercapnia model group), group II (disodium cromoglycate control group), group IV (curcumin treated group). The last 3 group rats were put in a hypoxia cabin where the concentrate of O2 was 8% - 11% and the concentrate of CO2 was 3% - 5%, for 8 h a day and lasting 4 w in total. Group III rats were intraperitoneally injected with disodium cromoglycate (20 mg/kg) and group IV rats were administrated with curcumin by gavage (150 mg/kg). The morphological changes of pulmonary vessel walls and the ultrastructure of mast cells were observed by the optics microscope and the transmission electron microscope. Mast cells and its degranulation state were measured by toluidine blue staining and immunohistochemistry. Data were expressed as means ± SD (standard deviation) and analyzed with SPSS17.0 software.</p><p><b>RESULTS</b>(1) By optics microscopy observation, the value of WA/TA was significantly higher in II group than other groups (P < 0.05). (2) Electron microscope showed that the endothelial cells of pulmonary arterioles in III and IV group were near to I group and the proliferation of pulmonary arterial media smooth cell layer and collagen fibers in adventitia was much lighter than those in II group. The membrane of mast cells was more intact in I, III, IV group than II group. (3) The number of mast cells, the degranulation rate of master cells and the number of positive tryptase stained cells in II group were significantly more than those in other groups. (P < 0.05).</p><p><b>CONCLUSION</b>Curcumin may inhibit the remodeling of pulmonary vessel induced by chronic hypoxia hypercapnia by mast cell regulation.</p>

Analysis of 90 autopsy cases in medical legal dispute / 法医学杂志

OBJECTIVE@#To analyze the significance of forensic autopsy in medical tangle.@*METHODS@#Ninety autopsy cases of medical legal dispute were retrospectively analyzed from the database of our department from 2001 to 2008. All cases were analyzed and classified based on age, sex, cause of death, clinic diagnosis and forensic diagnosis.@*RESULTS@#The age ranged from 1 day to 72 years, and the ratios of male to female is 1:1. The most common healthcare facilities involved were county hospitals (30 cases, 33.33%). The coincidence rate between clinical diagnoses and pathological diagnoses was 33.33%.@*CONCLUSION@#The forensic autopsy is valuable to solve or even avoid the occurrence of medical legal dispute.

Effects of curcumin on malondialdehyde and c-fos protein in hypoxia ischemia brain tissue in rats / 法医学杂志

OBJECTIVE@#To explore the effects of curcumin on the content of malondialdehyde (MDA) and the expression level of c-fos protein following hypoxia ischemia brain damage (HIBD) in rats.@*METHODS@#Sprague-Dauley (SD) rats were randomly divided into four groups as the following: sham group, hypoxia ischemia brain damage group, curcumin group and solvent control group. The content of MDA in the brain was measured by colorimetry. The expression level of c-fos protein in the cortex tissue was detected by immunohistochemistry. Morphologic and structural changes of neuron cells of the cortex were observed by electron microscopy.@*RESULTS@#The content of MDA was clearly lower in curcumin group than that in the other groups at the same time after HIBD. The expression level of c-fos protein was higher in the curcumin group than that in the other groups (P<0.05). Electron microscopy showed that the morphologic and structural changes of neuron cells of cortex in the curcumin group were reduced.@*CONCLUSION@#Curcumin could significantly decrease the content of MDA, increase the expression level of c-fos protein and reduce the damage of the neuron cells.

Acute necrotizing pancreatitis and postmortem autolysis of pancreas / 法医学杂志

OBJECTIVE@#To compare the pathomorphologic changes between the pancreas in acute necrotizing pancreatitis (ANP) and that in acute deaths of rats (within 48 hours) so as to find the distinctions.@*METHODS@#The animal models of ANP and other acute deaths (electroshock, mechanic asphyxia/strangle, and acute poisoning with tetramine) were established according to the criteria. Half-quantitative grading and image quantitative analysis methods were employed to observe the gross and microscopic changes of the pancreases.@*RESULTS@#Three features including inflammation infiltrate, fat necrosis and calcium deposit in the ANP group were considerably different from that in other acutely died rat group (P<0.05).@*CONCLUSION@#Inflammation infiltrate, fat necrosis and calcium deposit are the most important pathologic features found in ANP by common light microscope, distinguishing ANP from postmortem pancreatic autolysis.

Application of diatom detection using knead pulp method / 法医学杂志

OBJECTIVE@#A novel technology for detection of diatom was discussed.@*METHODS@#Five grams of testing sample were taken and the organics were removed using simple mechanical knead pulp method. The homogenized samples were concentrated by centrifugation, smeared, and then examined under light microscope.@*RESULTS@#Except for a few feather's grains, the vast majority of diatom could be identified easily with clear diatom striations. The organic diatom could also be easily detected by this methodology.@*CONCLUSION@#The detection of diatom using knead pulp method is not only simple and inexpensive with a higher successful rate, but also causes nearly no harm to human and environment.

Changes of the brain NSE, S100 and effect of ligustrazine in rats of chronic hypoxia and hypercapnia / 中国应用生理学杂志

<p><b>AIM</b>To investigate the changes of the brain NSE, S100 and ultrastructure and effect of ligustrazine in rats of chronic hypoxia and hypercapnia.</p><p><b>METHODS</b>Thirty rats were randomly divided into three groups: control group (A), hypoxia hypercapnia group (B), hypoxia hypercapnia added ligustrazine group (C). The brain NSE, S100 and ultrastructure were observed in rats using the technique of immunohistochemistry and electronic microscope.</p><p><b>RESULTS</b>(1) The mPAP was significantly higher in rats of group B than that of group A and it was much lower in rats of group C than that of group B. Differences of mCAP were not significant in three groups. (2) Serum NO of group B was significantly lower than that of group A, Serum NO of group C was higher than that of group B. (3) Immunohistochemistry showed the average value of integral light density (LD) of NSE and S100 was significantly much lower in rats of group B than that of group A and it was higher in rats of group C than that of group B. (4) The neuron and astrocyte of group B showed vacuolar degeneration and the myelin sheath showed separate. Damage of neuron is alleviated in rats of group C.</p><p><b>CONCLUSION</b>The hypoxia hypercapnia could induce damage of neuron and astrocyte in rats. The ligustrazine may be useful in protecting against hypoxia hypercapnia brain damage.</p>

Influences of perindopril on pulmonary vascular remodeling in normobaric hypoxia-hypercapnia rats / 中国临床药理学与治疗学

Aim To investigate the effects of perindopril, an angiotensin converting enzyme inhibitor, on mean pulmonary arterial pressure(mPAP) and pulmonary vascular remodeling.Methods Using the normobaric hypoxia-hypercapnia rat model,the changes of plasma angiotensin Ⅱ (Ang Ⅱ), mPAP and ultrastructure of pulmonary arteriolar wall were observed after administration of perindopril.Results The mPAP and AngⅡ were significantly greater and the ultrastructure of the small lung vessels had a more obvious change in the 4 weeks hypoxia-hypercapnia group than those in the control group.It was also found that perindopril decreased AngⅡ, declined mPAP and ameliorated vascular ultrastructure change.But the change of ultrastructure was similar in the 8 weeks hypoxia-hypercapnia group and perindopril treatment group.Conclusion It is suggested that chronic hypoxia-hypercapnia should induce the remodeling of pulmonary arteriolar structure via AngⅡ and that perindopril could ease pulmonary vascular remodeling in early stage.