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.

IL-17,as a pro-inflammatory cytokine,is one of the early initiation factors of the inflammato-ry response induced by T cells.It can induce and regulate multiple immune responses.Recent studies have re-vealed that myeloid neutrophils,macrophage,mast cell and other innate immune cells all can secrete large a-mount of early responding IL-17 in organs suffering ischemia/hypoxia,and in turn can activate,amplify and re-cruit neutrophils to the reperfusion-damaged tissue to release large amount of free radicals and lysozyme that cause IRI.Researchers have also provided evidence that appropriate administration of anti-IL-17 mono-antibody to neutralize IL-17 during early reperfusion stage would reduce the tissue damage.The purpose of this review is to summarize the research progress of the effects of IL-17 produced by innate immunocyte on organ reperfusion injury.

Objective To develop a rapid and sensitive method for the determination of bisphenol A in drinking water by liquid chromatography-electrospray tandem mass spectrometry. Methods Bisphenol A was extracted from the drinking water sample by a SEP-PAK C18 column,eluted with methanol and concentrated with a rotary evaporator. The extract was analyzed by liquid chromatography-tandem mass spectrometry with electrospray ionization. Results The calibration curve of bisphenol A was linear in the range of 5 -100 ng/ml,the linear equation was y =6 796.61x -8 655.64 and the correlation coefficients of linear calibration curve was 0.999 2. The limit of detection (S/N=3) and the limit of quantification (S/N=10) of bisphenol A were 0.007 5 ng/ml and 0.025 ng/ml,respectively. The rates of recovery of bisphenol A were from 83.46% to 94.00% and the relative standard deviation was 3.06% -4.60%. Conclusion The method is simple,rapid,accurate and is applicable to the qualitative and quantitative determination of bisphenol A in drinking water.

Focal adhesion kinase(FAK)and extracellular signal-regulated protein kinase(ERK),which are thought to play major roles in transducing surface receptor-derived signals into nucleus,are pivotal signal transduction molecules in mammalian cells.FAK is activated through phosphorylation by extracellular stimuli.p-FAK activates the downstream of ERK which changes cell action.FAK-ERK signal conduction factors are involved in a variety of biological responses,such as cell proliferation,differentiation,stress,apoptosis and malignant change.The biologic characters of FAK-ERK and advances in the study on its roles in tissue damage are reviewed in this article,which may provide a more effective way for estimation of wound age in forensic medicine.

To explore the applicability of interleukin 8(IL 8)to determination of wound age in the incised and stabbed human skin samples,an immunohistochemical study on the temporal expression of IL 8 was performed on 52 human skin wounds at different posttraumatic intervals.Expression of IL 8 was detectable in polymorphonuclear cells(PMNs)in the wound specimens aged 4h.In the wound specimens aged from 12～24h after injury,IL 8 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 IL 8 positive cells.Morphometrically,the ratio of the number of the IL 8 stained PMNs,MNCs and FBCs to total number of the cells in the wounds was evaluated and calculated.The ratio of the IL 8 positive cells was low in the wound specimens aged 4 and 6h(16 0?10 1%),and maximized in the wound specimens aged between 1 and 4 days(59 6%?8 7%).Thereafter,the ratio decreased and minimized in the specimens aged from 19 to 21 days(27 5?5 9%).The results suggest that IL 8 can be used as a marker for the wound age determination in both incised and stabbed human skin.

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 analysis the clinical feaures and risk factors of recurrent intracerebral hemorrhage(ICH). Methods Analysis clinical data of 68 recurrent ICH patients and compared with those of patients with first-ever ICH. Results 43 recurrences were seen in 38 patients.The mean interval between 2 episodes was(3.9?2.8)years.The most common type of recurrence was"ganglionic-ganglionic",next was"ganglionic-thalamic",and"ganglionic-lo- bar"seldom.Most recurrences occurred contrala-terally and seldom at the same site as the first episode.In contrast to those with first-ever ICH,patients with recurrent ICH most have a hypertension and course of longer,all with a poor control.Analysis showed that the hypertension course was positively and the age of the first hemorrhage was negatively associated with the recurrence of ICH.Conclusion The risk of recurrence of ICH is relatively high within the first 4 years after the previous hemorrhage.Positively control hypertension to normal level,It will have impartant meaning to prevent occurrence of recurrent ICH.

Objective To discuss the effect of expanded deltopectoral skin flaps in repairing scar contracture deformity of facio-cervical .Methods Twenty-four patients with scar contracture deformity of faciocervical were included into the study .We expanded the deltopectoral skin flap with dilator .8-10 weeks later ,the deltopectoral skin flap were transferred after resection of faciocervical scar .3-4 weeks later ,the pedicles were cut off ,and then the remaining wound were repaired .Results They had not exudation .Ex-pander exposure ,local hematoma and local infection respectively were 1 ,3 ,4 cases ,all did not affect operation effect by symptomatic treatment ,and obtain good postoperative effect .Deltopectoral flap was for 27 cases ,blood supply obstruction low temperature and skin flap necrosis respectively were 4 ,2 ,1 cases ,all get good healing after corresponding .Followed up for 0 .5 year ,all flaps were survived well ,no ectropion or mental cervical adhesion ,its color and texture were similar with surrounding normal skin appearance , so they were got high satisfaction .Conclusion Application of expanded deltopectoral skin flaps to repair scar contracture deformity of faciocervical can achieve good effect and may be an ideal method to repair ,which is worthy of clinical application .

Objective To investigate the experience of microinvasive surgical occlusion of ventricular septal defect (VSD).Methods A total of 142 children with VSD was given microinvasive surgical occlusion from March 2009 to December 2013 at our hospital.There were 90 males and 52 females,the age ranged from 8 months to 11 years,and body weight from 7 kg to 35 kg,and ventricular septal defects were divided into membranous type,film cycle headquarter type,pulmonary valve type,and muscle type.The diameter of VSD was 2 ～ 10 mm.Under general anesthesia,an incision was made in the lower part of sternum or intercostals space,and a special occluder was inserted to close the ostium via right ventricle puncture under the guidance of transesophageal echocardiography.Results A total of 139 cases had successful occluded with a 97.8％ of successful rate,using blocking umbrella 4 ～ 12#,including 25 eccentric umbrella.Two cases were operated under extracorporeal circulation because of aggravated aortic valve insufficiency.One case without handled muscular ventricular septal defect combined atrial septal defect for guide wire pass muscular defect failed.Full set did not have death and third degree A-V block.Conclusions Microinvasive surgical occlusion is easy to handle,operation-time short,and relative broad for the closure of ventricular septal defect.It has a fast recovery and good effectiveness with a beautiful outlook and safety.

Effects of glucose concentration and pH\|control upon production of exopolysaccharides of Grifola frondosa was studied.The results showed that glucose at concentration of 5% was favorable to exopolysaccharides production,and the pellets benefited from the controlled start pH as well as the controlled final pH of 3 5～4 0 to produce exopolysaccharides.Attempt on the exopolysaccharides production with recycled pellets in the saccharoid solution was also made in this work.