Objective To evaluate the influence of different antigen retrieval methods and pH values of fixatives on immunostaining of incised wound skin.Methods 3d-post-incision mouse skin were sampled and fixed in 4% formalin buffered with PBS at pH 5.0,6.0,7.0,8.0 and 9.0.The deparaffinized sections were treated 0.01mmol citrate buffuer(pH 6.0)in high pressure cooker or by microwave heating,or with pepsin,trypsin or microwave+pepsin.Thereafter,immunostainings of caspase-3,caspase-6,caspase-7,IL-8,and IL-10 were evaluated by immunohistochemical SABC method with ratio of immunostaining-positive cells,respectively.Results The immunostaining-positive cell ratios in sections treated by heating retrieval was higher than by proteinase treatment.The positive cell ratio was slightly higher by high pressure than by microwave heating.Sections were more easier peeled off during high pressure treatment than during micrwave heating.Sections treated with pepsin were fallen off easier than with trypsin digestion.The highest positive cell ratio was detected in sections by microwave+trypsin treatment without section peeling off among all antigen treatment methods.The best result was obtained in sections fixed in buffered formalin at pH 7.0 and 8.0.Conclusion The best immunostaining result may be obtained in mouse incised wound sections fixed in buffered formalin at pH 7.0 and 8.0 with microwave+trypsin treatment.

Hitherto, it is reported that caspase family includes 14 members at least, which play important roles in executing cell apoptosis. Experimental studies have proved that caspases are expressed and activated in the injured skin and during skin wound healing, which suggests that cell apoptosis induced by caspases may occur. Further investigation on the roles that caspases play in the injured skin and during skin wound healing may offer a new way for the clinical treatment of the skin wound and the medico-legal wound age determination. The article reviews the biologic characters of caspases and the advances in the studies on the caspases in the skin wound and wound healing. It is suggested that caspases exert significant parts during skin wound healing, which are worthy of further investigation.

Objective To investigate the expressions of caspase-6,-7 during skin wound healing and the applicability of the time-dependent expressions of caspase-6,-7 to determination of the wound age.Method The expressions of caspase-6,-7 were studied in skin incised wound in mice by immunohistochemical SABC method,and the non-incised mouse skin was used as control.Results Expressions of caspase-6,-7 were detectable in polymorphonuclear cells (PMNs) in the wound specimens aged 6h. In the wound specimens aged from 12h to 24h after injury, caspase-6,-7 were identified in a large number of infiltrating PMNs and part of mononuclear cells (MNCs). Afterwards, the MNCs and fibroblastic cells (FBCs) accounted for the most part of the caspase-6,-7-positive cells. Morphometrically, the ratios of the number of the caspase-6,-7-stained PMNs, MNCs and FBCs to total number of the cells in the wounds were evaluated and calculated. The ratios of the caspase-6,-7-positive cells were low in the wound specimens aged from 0h to 3h, and maximized in the wound specimens aged 3 day. Thereafter, the ratios decreased and minimized in the specimens aged 14 day.Conclusion The results suggest that caspase-6,-7 were expressed in neutrophils, macrophages and fibroblasts during healing process of skin incised wound in mice. Caspase-6,-7 may be used as a marker for the wound age determination.

OBJECTIVE: To understand the assessment on the criminal responsibility of drug-induced mental disorders and judicial experts' opinions. METHODS: The judicial experts from institutes of forensic psychiatry in Shanghai were selected. They were asked to finish a self-made questionnaire of assessment on the criminal responsibility of drug-induced mental disorders by letters and visits. RESULTS: Most of experts knew the special regulation, "not suitable for evaluation" towards the criminal responsibility of drug-induced mental disorders of the guideline promulgated by Ministry of Justice. Before and after the guideline was issued, no expert made a no-responsibility opinion in such cases. After the guideline was issued, some experts made a full-responsibility or limited-responsibility opinion in such cases. There was a little disagreement among the experts in the case that the crime was unrelated with mental symptoms or the criminals used drugs even though he knew it could induced insanity. But there were still many obvious disagreements among experts in the case that crime was related to such symptoms and person was no ability to debate. Most experts agreed to settle the disagreements with improved legislative perfection. CONCLUSION Most experts are not strictly complying with the assessment guidelines during their practice, and there is still an obvious disagreement towards the criminal responsibility of drug-induced mental disorders.
China ; Crime/psychology* ; Criminals/psychology* ; Data Collection ; Expert Testimony ; Forensic Psychiatry ; Humans ; Liability, Legal ; Male ; Mental Competency ; Mental Disorders/psychology* ; Psychotic Disorders ; Surveys and Questionnaires

ObjectiveTo investigate the effects and mechanism of rosuvastatin on the expression of tissue factor in cultured human monocyte-macrophages cells which was induced by oxidized low density lipoprotein(ox-LDL).MethodsThe human monocyte-macrophages cells were divided into four groups:control group,ox-LDL group,Poly-inosine monophosphate (Poly Ⅰ) group,rosuvastatin group.The expression of LOX-1mRNA and TF mRNA was assayed by RT-PCR.The ELISA was performed to determine the protein concentration of TF.ResultsCompared with control group,the expression of LOX-1 mRNA and TF mRNA was increased in the ox-LDL group[ (3.25156±0.15772) vs (1±0) ; (2.522451±0.138967) vs (1±0) ],and it was in a concentration-dependent manner (P＜0.01).Compared with the expression of LOX-1 mRNA in the Poly-inosine monophosphate group and rosuvastatin group,TF mRNA were decreased in the ox-LDL group[ (2.95139±0.157253) vs(3.25156±0.15772) ; (2.877343±0.156558) vs(3.25156±0.15772) ; (1.811956±0.169699) vs (2.522451±0.138967) ; (1.687701±0.174647) vs (2.522451±0.138967)],and it was in a concentration-dependent manner(P＜0.05).Compared with control group,the expression of TF in the ox-LDL group was increased [(207.7233±1.154701) vs (184.8467±0.871799) ],and it was in a concentration-dependent manner (P＜0.01).Compared with the Poly-inosine monophosphate group and rosuvastatin group [(197.8733±1.505003) vs (207.7233±1.154701) ;(202.9567±2.722744)vs(207.7233±1.154701) ],the expression of TF in the ox-LDL group were decreased,and it was in a concentration-dependent manner (P＜0.05).ConclusionsLOX-1 may be responsible for the expression of TF in human monocyte-macrophages cells induced by ox-LDL.Rosuvastatin is able to down-regulate the expression of LOX-1mRNA in human monocyte-macrophages cells through oxLDL,and TF mRNA and TF expression can be reduced.

Objective To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle me-chanical injury in rats. Methods The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury ) and con-trol group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. Results At post-injury 6-12 h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN de-creased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14 d, the percentage of FBC reached peak. Conclusion The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.

0.05). Conclusions Silencing STAT3 gene can decrease STAT3 gene expressions , increase Bax gene expressions , induce cell apoptosis and suppress the tumor growth in the human breast carcinoma model by the RNAi technology .

Objective To develop a software for diabetic dieting guide.Method One set of food catering software for diabetic patients was developed based on the energy exchange among a variety of foods using VEB on conditions of a balance of total energy and three major energy material.Result The food catering software was feasible and reasonable so that it could control the indexes of blood glucose,blood lipid and body mass index(BMI).Conclusion The food catering software for diabetic patients is perfect in function and can enhance diabetic compliance and feasiblility.

Objective To study the effects of Toll-like receptor 4(TLR4) on oxidized low density lipoprotein ( ox-LDL) induced macrophage apoptosis and its possible mechanism .Methods THP-1 derived macrophages were divided into four groups including untreated control group , ox-LDL treated group , ox-LDL+LPS treated group and tunicamycin treated group .MTT assay and flow cytometry analysis were performed to measure cell vitality and cell apoptosis , respectively .Oil red O staining was used to observe the phagocytosis of lipids by macrophages .The persistent and intense endoplasmic reticulum ( ER) stress markers were de-tected by analyzing the expression of glucose-regulated protein 78 ( GRP78 ) and CCAAT/enhancer-binding protein homologous protein ( CHOP) at mRNA and protein levels by q-RT-PCR and Western blot .Small in-terfering RNA ( siRNA) was used to silence the expression of TLR 4 to further elucidate its possible mecha-nism.Results Flow cytomotry and MTT assay showed that the number of apoptotic cells in ox-LDL+LPS treated group were increased more significantly than that in ox-LDL treated group (P<0.01), and cell apop-tosis in both two groups were greater than that in control group (P<0.01).Compared with control group, the expression of GRP78 and CHOP at mRNA and protein levels were up-regulated in ox-LDL+LPS treated group and ox-LDL treated group (P<0.01), and the expression of GRP78 and CHOP in ox-LDL+LPS treated group was significantly higher than that in ox-LDL treated group (P<0.01).Silenced expression of TLR4 al-leviated the endoplasmic reticulum stress (P<0.05).Conclusion Increased expression of CHOP contribu-ted to cell apoptosis .TLR4 might promote ox-LDL induced macrophage apoptosis through accelerating endo-plasmic reticulum stress .

OBJECTIVE: To investigate the expression of caspase-3 during skin wound healing and explore the applicability of caspase-3 to determination of wound age. METHODS: The expression of caspase-3 were performed on 33 mice skin incised wounds at different posttraumatic intervals and 3 mice as control by immunohistochemical technique. RESULTS: Expression of caspase-3 was detectable in polymorphonuclear cells (PMNs) in the wound specimens aged 6 h. In the wound specimens aged from 12 h to 24 h after injury, caspase-3 was identified in a large number of infiltrating PMNs and part of mononuclear cells (MNCs). Afterwards, the MNCs and fibroblastic cells (FBCs) accounted in the most part of the caspase-3 positive cells. Morphometrically, the ratio of the number of the caspase-3 stained PMNs, MNCs and FBCs to total number of the cells in the wounds was evaluated and calculated. The ratio of the caspase-3 positive cells was low in the wound specimens aged from 0 h to 3 h (4.53 +/- 6.53)%, and maximized in the wound specimens aged 3 day (62.66 +/- 4.84)%. Thereafter, the ratio decreased and minimized in the specimens aged 14 day (21.56 +/- 4.54)%. CONCLUSION The results suggest that caspase-3 may play an important role in inducing neutrophil, macrophage and fibroblast apoptosis during skin wound healing. Furthermore, caspase-3 may be used as a marker for the wound age determination.
Animals ; Apoptosis ; Caspase 3 ; Caspases/biosynthesis* ; Female ; Immunohistochemistry ; Male ; Mice ; Skin/metabolism* ; Staining and Labeling ; Time Factors ; Wound Healing ; Wounds and Injuries/physiopathology*