Objective To optimize the culture medium for scarless wound healing in outbred fetal Kunming (KM) mice,and to locate and preliminarily analyze the region with active cell proliferation.Methods After 6 pregnant mice were sacrificed at the gestational age of 15 days,a total of 180 skin grafts were obtained from the back of 60 fetal mice,and wounds on the skin grafts were made uniformly with minimally invasive sterile device.Then,these skin grafts with wounds were randomly and equally divided into 6 groups to be treated with high-glucose Dulbecco's modified Eagle's medium (DMEM),DMEM + 5％ fetal bovine serum (FBS),DMEM + 10％ FBS,low-glucose Eagle's minimum essential medium (MEM),MEM + 5％ FBS,and MEM + 10％ FBS for 3 days,respectively.Then,these skin grafts were embedded in optimal cutting temperature (OCT) compound,and subjected to hematoxylin and eosin (HE) staining to select the optimal culture condition for wound healing.Under the optimal culture condition,the region with active cell proliferation was located by labeling and tracking cutaneous stem cells with 5-ethynyl-2-deoxyuridine (EdU),and activator protein-1 (AP-1) expression was detected by immunofluorescent staining.Results After 3-day cultivation,wound healing rates significantly differed among the DMEM group,DMEM + 5％ FBS group,DMEM + 10％ FBS group,MEM group,MEM + 5％ FBS group and MEM + 10％ FBS group (0,3.33％,6.67％,3.33％,46.67％ and 26.67％,respectively,x2 =41.39,P ＜ 0.05).Additionally,the MEM + 5％ FBS group showed a significantly higher wound healing rate than the other groups except the MEM + 10％ FBS group (all P ＜ 0.01).Logistic regression analysis showed that the type of basal medium (MEM:OR =11.717,95％ CI:3.274-41.934,P ＜ 0.001) and FBS concentrations (5％ FBS:OR =24.625,95％ CI:3.027-200.299,P =0.003;10％ FBS:OR =13.449,95％ CI:1.618-111.813,P =0.016) were factors influencing fetal wound healing.Under the culture condition of 5％ FBS + MEM,the dermis and epidermis healed well without epidermal thickening.The cutaneous stem cell labeling technique showed that the papillary dermis and basal layer of the epidermis were the two major regions with active cell proliferation in the wound of fetal mouse skin.Moreover,AP-1 was expressed abundantly in these two regions as well.Conclusions The culture condition of 5％ FBS + MEM is considered to be optimal for in vitro wound healing in fetal mice.The papillary dermis and basal layer of the epidermis are two major regions with active cell proliferation during wound healing in fetal mouse skin,and play important roles in wound healing process.

Objective To observe the changes of brainstem auditory evoked potential (BAEP) and lower limb short latency somatosensory evoked potential ( SLSEP) in acute carbon monoxide poisoning patients and explore the predictive value of BAEP and SLSEP for delayed encephalopathy after acute carbon monoxide poisoning (DEACMP). Methods Sixty-six acute CO poisoning patients was divided into a DEACMP group and a non-DEACMP group according to their symptoms. At the same time, 60 healthy subjects were included in the control group. BAEP and lower limb SLSEP dynamic testing were conducted at different time points. Results The abnormal rates in the first test of BAEP and lower limb SLSEP in the acute CO poisoning patients were 31.8% and 68.2% respectively.The BAEP results in the acute stage and later in the DEACMP group were comparable to those of the non-DEACMP group and the normal control group. Interpeak latency of Ⅰ-Ⅲ was not significantly different, but the remaining indices were significantly longer. Compared to the control group, the lower limb SLSEP readings in the acute DEACMP group and the non-DEACMP group were significantly longer. Compared to the non-DEACMP group and the control group, the SLSEP results of the DEACMP group were significantly extended. The specificity index of the BAEP predicted DEACMP up to 85.7% , significantly more than SLSEP testing. Conclusions Early detection of BAEP can help predict DEACMP. This finding should be promulgated and applied.

【Objective】 To compare the desalination effects of five desalination methods and their effects on the components for human coagulation factor Ⅷ(FⅧ), and provide reference for selection of protein desalination methods. 【Methods】 Sephadex G-25 Medium gel, Fractogel EMD BioSEC gel, ultrafiltration, room temperature dialysis and 4℃ dialysis were used to desalt human FⅧ. The desalination effect was evaluated by the removal rate of Na +, citrate ion and glycine. FⅧ protein recovery, FⅧ activity (FⅧ∶C), VWF antigen (VWF∶Ag), VWF activity(VWF∶Ac), VWF polymers and SDS-PAGE analysis before and after desalination were compared to evaluate the effect of desalination on FⅧ components. 【Results】 In terms of desalination effect, the removal rate of Na⁺ was the lowest in ultrafiltration desalination, while that of Fractogel EMD BioSEC gel was the highest [(97.90±0.06) % vs (99.82±0.07) %]. Except that there was no statistical significance between Sephadex G-25 Medium gel desalination and Fractogel EMD BioSEC gel desalination (P=0.90), the removal rates of the other four methods were statistically significant. The removal rate of glycine was the lowest in ultrafiltration desalination, wihle that of Fractogel EMD BioSEC gel desalination was the highest [(95.78±0.42) % vs (99.81±0.08) %]. Significant difference in glycine removal was noticed in ultrafiltration desalination, but not among the other four desalination methods. There was no significant difference in the removal rate of citrate ions among the five methods (P=0.85). For the effect of FⅧ components, FⅧ∶C, VWF∶Ag, VWF∶Ac and protein recovery rates of ultrafiltration desalination were the highest, with (18.34±1.99) IU/mL, (11.81±0.33) IU/mL, (12.26±0.58) IU/mL and (97.13±1.37) %, respectively. There was no significant change in VWF∶Ac/VWF∶Ag before and after desalination by the five methods. SDS-PAGE and VWF polymer analysis showed that different desalination methods had no significant impact on protein composition. 【Conclusion】 Although different desalination methods had no significant effect on the composition of FⅧ protein, the desalination effect was different. Moreover, different desalination methods had significant effects on protein recovery, FⅧ∶C, VWF∶Ag and VWF∶Ac. The selection of desalination methods should be more considered during protein processing,