Objective To investigate Smac/DIABLO and cytochrome c(cyt-c)mRNA levels in liver tissue of rats with acute hepatic failure treated by microencapsulated hepatocyte.Methods Acute hepatic failure were induced by intraperitoneal injection of D-galactosamine in rats.and the rats were treated with microencapsulated hepatocytes,free hepatocytes and physical saline(contr01),respectively.Smac/DIABLO and cyt-c mRNA in liver tissue was detected by RT-PCR and the mRNA expression levels among three groups were compared.Results Smac/DIABLO and cyt-c mRNA levels in liver tissues of rats with acute hepatic failure were higher than those of normal rata(F=4.345,14.821,47.565,42.178 and 62.961,P<0.05).The peak values of Smac/DIABLO and cyt-C mRNA expressions in free hepatocytes and control groups were at 48 h.while that in microencapsulated hepatoeytes group was at 24 h.Conclusion Smac/DIABLO and cyt-c mRNA expression is an indicator of apoptosis of hepatocytes.

Objective To investigate the clinical and laboratory features of severe cases of hand,foot and mouth disease (HFMD) in Wenzhou, Zhejiang Province. Methods Clinical data of 107 children with HFMD, including 97 severe and 10 critical cases treated in Children' s Hospital Affiliated to Wenzhou Medical College during January and May 2010 were retrospectively analyzed. One hundred and fifty children with mild HFMD were also selected as the controls. Clinical features and laboratory results were compared between the two groups. Results Fever, rash and infection in central nervous system were observed in all patients with severe HFMD, and symptoms on respiratory, digestive and cardiovascular systems were more serious than those of mild HFMD cases. White blood cell counts (WBC) were higher in severe group than those in controls (t = 12.72, P ＜0.01). Hyperglycemia (9. 2 mmol/L) and abnormal troponin (0. 3 -9. 0 ng/mL) were presented in all the critical patients. Cerebrospinal fluid WBC counts were raised in 97 severe HFMD patients (98.5 × 106/L for average) with predominance of lymphocytes. Among 107 severe patients, EV71 was positive in 70, including all 10 critical cases. Conclusion Involvement of nervous,respiratory and digestive symptoms is common in severe cases of HFMD, and EV71 is the predominant pathogen.

Objective:To study the possible immune escape mechanisms of HCoV-OC43 from human dendritic cells(DC).Methods:HCoV-OC43 was isolated from clinical specimen using BSC-1 cells and identified by Real-time PCR,and the cytopathic effect was observed by phase contrast microscope.DCs were induced in vivo using hu-GM-CSF and IL-4 cytokines,and after 7 days of differentiation,DCs were infected by HCoV-OC43.The morphology of HCoV-OC43 infected DC was observed by transmission electron microscope,and the cytokines related to DC functions were detected by Real-time PCR after infection.DC proportion and function related co-stimulatory molecules were analyzed by flow cytometry.Results:In vitro HCoV-OC43 infected human DC model was successfully built.HCoV-OC43 can infect DC and generate immune response of DC in vitro,but no virus nucleonic acid could be detected in culture supernatant.The DC expression of IFN-α,IFN-β,CCL3 and CCL5 were significant decreased when infected with HCoV-OC43,but the expression of costimulatory molecules including HLA-DR,CD1c and CD86 were not affected by HCoV-OC43 infection.Conclusion:Human DC could be infected by HCoV-OC43 and generate immune response,but could not produce progeny virus.HCoV-OC43 may escape from immune response by suppressing the expression of IFN-α and other inflammatory cytokines and chemokines in DC.

Objective To analyze the relationship between the level of serum calcium binding protein S 100A12 and the severity of acute pancreatitis (AP) ,so as to provide a simple and reliable method for judging the severity of AP .Methods A total of 68 pa-tients with AP from January 2015 to January 2016 in Luzhou Hospital of Traditional Chinese Medicine were selected as objects in this study ,and complete the examination after diagnosis ,modified CT severity index (MCTSI) was used to grade the severity ,serum calcium binding protein S100A12were tested and compared in patients with different disease grading .The ROC curve of different se-verity patients were made .Judging the severity of critical value according to the S 100A12 curve inflection point ,basing on the judg-ment of MCTSI ,the sensitivity ,specificity and accuracy of S 100A12 in grading condition of AP were calculated ,then the value of S100A12 was evaluated .Results The level of S100A12 increased with MCTSI grade rising ,there were significant differences in S100A12 level between different grades patients(P<0 .05) .ROC curve analysis showed that the clinical limits of gradeⅠto Ⅲ were 26-80 ,81-260 ,>260 ng/mL respectively .The sensitivities accuracy of S100A12 judging grade Ⅰ ,grade Ⅱ and grade Ⅲ were 88 .89% ,94 .12% ,80 .00% ,the specificity were 75 .00% ,94 .12% ,96 .55% ,the accuracy were 82 .35% ,94 .12% ,94 .12% respec-tively .Conclusion The serum calcium binding protein S100A12 estimates is consistent with MCTSI for judging the severity condi-tion of AP .

Objective:To observe the efficacy and safety of oral administration of low-molecular-weight collagen peptide on facial skin rejuvenation.Methods:A total of 66 female volunteers in Department of Dermatology, Beijing Friendship Hospital, Capital Medical University aged from 35 to 50 years old (average 42.89±4.44) from November 2018 to February 2019 were enrolled and randomly divided into two groups, 33 cases each. The testing group was given 5 g Su Yan Xin Ji collagen peptide for 12 weeks; the control group was given 5 g Tai Duo Jian collagen protein powder for 12 weeks. The effects were evaluated and analyzed using VISIA and CK complexion analysis system before and after 1, 2, 4 , 12 weeks of oral administration collagen; meanwhile, the volunteers' overall satisfaction and adverse reactions were also recorded.Results:In total, 61 volunteers completed the study. After 12 weeks of oral administration, the skin hydration of testing group and control group were all increased (65.41±10.60 vs 59.82±9.26), the transepidermal water loss, wrinkles, textures, pores, red areas and porphyrin were all decreased (19.19±4.24 vs.21.50±5.10; 7.38±3.67 vs. 8.98±6.67; 5.55±3.07 vs.6.60±4.84; 16.94±9.30 vs. 17.95±8.85; 21.92±4.60 vs. 22.11±5.34; 10.31±7.03 vs. 11.62±8.58). There were statistically significant differences between the 7 parameters before and after oral administration ( P<0.05). The difference of skin hydration between the testing and control group was statistically significant ( t=2.317, P=0.024). Although there was no statistical difference among the other six parameters, the improvement degree of the testing group was better than that of the control group. Surface spots, UV spots and brown spots of two groups showed no significant difference before and after treatment. Overall satisfaction of the testing group and the control group were 86.67% vs. 61.29% with significant difference 12 weeks after treatment ( χ2=5.074, P=0.024), while there were no significant differences in incidence of adverse reactions between two groups ( P>0.05). Conclusions:Oral administration of low-molecular-weight-collagen peptide can improve skin textures, moisturize skin with high overall satisfaction. It is an effective method of facial rejuvenation and better than that of collagen protein powder.

Humans ; Abdomen ; Dantrolene ; East Asian People ; Malignant Hyperthermia

Objective: To study the effect of stem cell factor (SCF) on the angiogenic ability of cocultured dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs). Methods : This study has been reviewed and approved by the Ethics Committee. The experiment was split into the HUVECs, SCF+HUVECs, DPSCs+HUVECs, and SCF+DPSCs+HUVECs groups. A mixture of SCF and culture medium was used to prepare a mixed culture medium with an SCF concentration of 100 ng/mL. In vitro coculture of DPSCs and HUVECs was performed at a 1∶5 ratio. CCK-8 proliferation assay was used to observe the proliferative capacity of cells in each group on days 1, 3, 5, and 7. Wound healing and Transwell migration assays were used to detect the effect of SCF on cell migration under either direct or indirect coculture conditions, respectively. In vitro angiogenesis experiments were performed to detect the angiogenic capacity of the cells in each group. The vascular endothelial growth factor A (VEGFA) concentration in the cell culture supernatant was detected using ELISAs, and the protein expression levels of CD31, CD34, and VEGFA were detected using Western blot analysis. Results : Wound healing and Transwell migration experiments showed that SCF significantly promoted the migration of cocultured DPSCs and HUVECs (P<0.05). The in vitro angiogenesis experiment showed that the number of branches and the total length of branches of tubular structures in the SCF+DPSCs+HUVECs group were significantly greater than those of the other groups (P<0.05), and the expression levels of the vascular-related proteins CD31, CD34, and VEGFA in this group were greater (P<0.01). Conclusion SCF can enhance the migration and in vitro angiogenesis of cocultured DPSCs and HUVECs.

Nonalcoholic fatty liver disease (NAFLD), especially nonalcoholic steatohepatitis (NASH), is a common hepatic manifestation of metabolic syndrome. However, there are no effective therapy to treat this devastating disease. Accumulating evidence suggests that the generation of elastin-derived peptides (EDPs) and the inhibition of adiponectin receptors (AdipoR)1/2 plays essential roles in hepatic lipid metabolism and liver fibrosis. We recently reported that the AdipoR1/2 dual agonist JT003 significantly degraded the extracellular matrix (ECM) and ameliorated liver fibrosis. However, the degradation of the ECM lead to the generation of EDPs, which could further alter liver homeostasis negatively. Thus, in this study, we successfully combined AdipoR1/2 agonist JT003 with V14, which acted as an inhibitor of EDPs-EBP interaction to overcome the defect of ECM degradation. We found that combination of JT003 and V14 possessed excellent synergistic benefits on ameliorating NASH and liver fibrosis than either alone since they compensate the shortage of each other. These effects are induced by the enhancement of the mitochondrial antioxidant capacity, mitophagy, and mitochondrial biogenesis via AMPK pathway. Furthermore, specific suppression of AMPK could block the effects of the combination of JT003 and V14 on reduced oxidative stress, increased mitophagy and mitochondrial biogenesis. These positive results suggested that this administration of combination of AdipoR1/2 dual agonist and inhibitor of EDPs-EBP interaction can be recommended alternatively for an effective and promising therapeutic strategy for the treatment of NAFLD and NASH related fibrosis.

Objective: To investigate the molecular evolution characteristics of human coronavirus (HCoV) subtypes in patients with fever and respiratory tract infection in Guangzhou from 2010 to 2012. Methods: Partial fragments of NP, RdRp and S genes of HCoV-OC43, HCoV-229E and HCoV-NL63 positive samples were amplified by RT-PCR and sequencing. Bioinformatics software, including Bio-edit, Mega4.0 and Clustal1.83 were used for comparison and analysis of NP, RDRp and S gene sequences. Molecular evolutionary tree of different gene regions of HCoV-OC43, HCoV-229E and HCoV-NL63 were built. Results: No remarkable variation or recombinant strain of HCoV-OC43, HCoV-229E and HCoV-NL63 was found in Guangzhou during 2010—2012. The HCoV-OC43 substrains were genetically closest to the strains found in Belgium and Hong Kong (GenBank accession number JN129834 and AY903460). HCoV-229E substrains were genetically closest to those found in Amsterdam (GenBank accession number JX503060) and HCoV-NL63 most genetically close to those in Amsterdam and Beijing (GenBank accession number JX104161 and DQ445911). The NP and RDRp genes of all subtypes were highly conserved, while S gene was more variable. Conclusions There were at least 3 substrains of HCoV-OC43, HCoV-229E and HCoV-NL63 epidemic in Guangzhou during 2010—2012, and no remarkable variation or recombinant viral strain was found. The NP and RDRp genes of all subtypes were highly conserved and can be used in virus detection, while S gene was more variable and suitable for phylogenetic and variation study.

Currently, there is still no effective curative treatment for the development of late-stage liver fibrosis. Here, we have illustrated that TB001, a dual glucagon-like peptide-1 receptor/glucagon receptor (GLP-1R/GCGR) agonist with higher affinity towards GCGR, could retard the progression of liver fibrosis in various rodent models, with remarkable potency, selectivity, extended half-life and low toxicity. Four types of liver fibrosis animal models which were induced by CCl₄, α-naphthyl-isothiocyanate (ANIT), bile duct ligation (BDL) and Schistosoma japonicum were used in our study. We found that TB001 treatment dose-dependently significantly attenuated liver injury and collagen accumulation in these animal models. In addition to decreased levels of extracellular matrix (ECM) accumulation during hepatic injury, activation of hepatic stellate cells was also inhibited via suppression of TGF-β expression as well as downstream Smad signaling pathways particularly in CCl₄-and S. japonicum-induced liver fibrosis. Moreover, TB001 attenuated liver fibrosis through blocking downstream activation of pro-inflammatory nuclear factor kappa B/NF-kappa-B inhibitor alpha (NFκB/IKBα) pathways as well as c-Jun N-terminal kinase (JNK)-dependent induction of hepatocyte apoptosis. Furthermore, GLP-1R and/or GCGR knock-down results represented GCGR played an important role in ameliorating CCl₄-induced hepatic fibrosis. Therefore, TB001 can be used as a promising therapeutic candidate for the treatment of multiple causes of hepatic fibrosis demonstrated by our extensive pre-clinical evaluation of TB001.