Objective To compare the effects of nitinol alloy memorial stent with silastic tube in treating the stenosis of the rabbit.Methods 16 rabbits with external ear canal(EEC) stenosis were randomly divided into two groups.One group was implanted with skin on the EEC wound while the other not.By self-comparison method nitinol alloy memorial stem was implanted in a rabbit's one ear and the silastic tube in the other.After days 5,15,30,and 60 later,the diameters of the external ear canal (with two materials planted) were measured respectively in the skin-planting group.In the naked group scar tissues were harvested and tested separately according to the planting material.HE coloration were used to study fibroblast hyperplasty while RT-PCR were applied to detecting the TGFβ1mRNA expression.The two brackets were compared according to their effect to scar hyperplasty.Results The EEC diameters using nitinol alloy memorial stem were found more spacious than using silastic tube.HE coloration showed the fibroblast hyperplasia was more mitigatory by using the nitinol alloy memorial stent.RT-PCR also found the TGFβ1mRNA expression was low by using same material.Conclusion The nitinol alloy memorial stent shows obvious superiority over silastic tube in external ear canal stenosis therapy.

Objective: To determine the changes of serum Tau protein, glial fibrillary acidic protein (GFAP), tumor necrosis factor alpha (TNF-α), and malonaldehyde (MDA) in rats after blast-related traumatic brain injury (BTBI) and to provide relative information for further studies on BTBI mechanism and seek specific biomarkers for BTBI.
Methods: Ninety male Sprague-Dawley rats were randomly assigned into three groups: control group, moderate blast injury group, and severe blast injury group (n=30 for each). Rats in the moderate and severe blast injury groups were respectively exposed to corresponding levels of BTBI. After explosion, serum levels of Tau, GFAP, TNF-α, and MDA in each group were determined by Elisa assay at different time points after injury (8 h, 24 h, 3 d, and 6 d). The extent of brain damage was detected by Nissl staining and TUNEL assay.
Results: Serum levels of Tau and GFAP rapidly increased and reached the peak at 24 h after either moderate or severe blast injury. All the values were significantly higher than control group at all time points (P<0.05). Serum TNF-α level of both injury groups peaked at 8 h after BTBI and stayed significantly higher than control group at all time points (P<0.05). Serum MDA of two injury groups began to significantly increase at 3 d and the level stayed significantly higher than control group until 6 d (P<0.05). Moreover, unlike the other biomarkers, serum MDA of severe blast injury group was significantly higher than moderate blast injury group at 6 d (P<0.05).
Conclusion: The changes of serum Tau, GFAP, and TNF-αshowed a good sensitivity at the acute phase after BTBI (within 24 h). However, their specificity and correlation with the extent of injury were limited in this experiment. Moreover, although the change of serum MDA showed a poor sensitivity and specificity to the diagnosis of BTBI during the first few days, it can reflect the injury degree at 6 d after injury. Therefore, further studies are needed to improve the methods of detecting more serum markers and investigate the significance of multiple markers in diagnosing BTBI.

Objective To express nonstructural protein 2 transactivated protein (NS2TP) of hepatitis C virus (HCV) in the prokaryotic expression system and prepare polyclonal antibody,and to study the expressions in different liver tissues.Methods NS2TP gene was amplified by polymerase chain reaction (PCR) technique and cloned into the prokaryotic expression vector pET-32a(+),which was transformed into E.coli BL21.The protein was induced by isopropyl thiogalactose (IPTG) and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and confirmed by Western blotting assay.The recombinant protein were expressed and purified in a large amount.The rabbit was immunized with the purified protein to prepare polyelonal antibody.The liver tissues of patients with chronic HCV infection and healthy controls were detected by immunohistochemistry method.Results The recombinant NS2TP protein (relative molecular mass:33×103 ) and polyclonal antibody with high titer and specificity were successfully prepared.NS2TP expressions in the liver of patients with chronic HCV infection were higher than those of healthy controls,and were mainly distributed in the nucleus of hepatocytes.Conclusions The NS2TP expression level and intracellular location in liver tissue of patients with chronic HCV infection are understanded,which could bring new clues for further study of the biological function of NS2TP and the pathogenesis of HCV infection.

Objective To investigate the changes of CD4+ CD25+ regulatory T lymphocyte (Treg) and expressions of folkhead helix transcription factor 3 (FoxP3) in intestinal mucosa in human immunodeficiency virus (HIV) infected patients. Methods Twenty-one HIV infected patients and 17 control subjects without HIV infection were included in this study. The expression of FoxP3, which was considered as a specific marker of CD4+ CD25 + Treg, was detected in intestinal mucosa specimens from HIV infected patients by immunohistochemistry. Meanwhile, the in situ expression of CD4+ T lymphocyte was also determined by immunohistochemistry. The data were analyzed by t test. Results The positive labeling index of CD4+ T lymphocyte in intestinal mucosa was significantly lower in HIV infected patients compared to the controls (11. 56%±4. 44% vs 43. 49% ±8. 90% ,t=-11. 86,P<0. 01). The positive labeling index of FoxP3 in intestinal mucosa was also significantly lower in HIV infected patients compared to the controls (0.46% ± 0.20% vs 1. 18% ± 0. 44% ,t= - 5. 98,P<0.01). Conclusion The depletion of CD4+ CD25+ Treg is accompanied with the depletion of CD4 + T lymphocyte and the reduction of FoxP3 expression in intestinal mucosa of HIV infected patients.

Objective To explore the effects of deletion of protein 4.1R on hepatocyte proliferation,apoptosis,and glycolysis and the molecular mechanisms.Methods A 4.1R-/-HL-7702 cell line was constructed using CRISPR/Cas9 technique,and with 4.1R+/+HL-7702 cells as the control,its proliferative capacity and cell apoptosis were assessed using CCK-8 assay,EdU-488 staining,flow cytometry and Annexin V-FITC/PI staining at 24,48,72 h of cell culture.The changes in glucose uptake,lactate secretion,ATP production and pH value of the culture supernatant of 4.1R-/-HL-7702 cells were determined.The mRNA expressions of the key regulatory enzymes HK2,PFKL,PKM2 and LDHA in glycolysis were detected with qRT-PCR,and the protein expressions of AMPK,p-AMPK,Raptor and p-Raptor were determined using Western blotting.Results Western blotting and sequencing analysis both confirmed the successful construction of 4.1R-/-HL-7702 cell line.Compared with the wild-type cells,4.1R-/-HL-7702 cells exhibited a lowered proliferative activity with increased cell apoptosis.The deletion of protein 4.1R also resulted in significantly decreased glucose uptake,lactate secretion and ATP production of the cells and increased pH value of the cell culture supernatant.qRT-PCR showed significantly decreased mRNA expressions of the key regulatory enzymes in glycolysis in 4.1R-/-HL-7702 cells.Compared with those in HL-7702 cells,the expression levels of AMPK and Raptor proteins were decreased while the expression levels of p-AMPK and p-Raptor proteins increased significantly in 4.1R-/-HL-7702 cells.Conclusion Deletion of protein 4.1R in HL-7702 cells results in reduced proliferative capacity,increased apoptosis and suppression of glycolysis,and this regulatory mechanism is closely related with the activation of the downstream AMPK-mTORC1 signaling pathway.

Objective To explore the effects of deletion of protein 4.1R on hepatocyte proliferation,apoptosis,and glycolysis and the molecular mechanisms.Methods A 4.1R-/-HL-7702 cell line was constructed using CRISPR/Cas9 technique,and with 4.1R+/+HL-7702 cells as the control,its proliferative capacity and cell apoptosis were assessed using CCK-8 assay,EdU-488 staining,flow cytometry and Annexin V-FITC/PI staining at 24,48,72 h of cell culture.The changes in glucose uptake,lactate secretion,ATP production and pH value of the culture supernatant of 4.1R-/-HL-7702 cells were determined.The mRNA expressions of the key regulatory enzymes HK2,PFKL,PKM2 and LDHA in glycolysis were detected with qRT-PCR,and the protein expressions of AMPK,p-AMPK,Raptor and p-Raptor were determined using Western blotting.Results Western blotting and sequencing analysis both confirmed the successful construction of 4.1R-/-HL-7702 cell line.Compared with the wild-type cells,4.1R-/-HL-7702 cells exhibited a lowered proliferative activity with increased cell apoptosis.The deletion of protein 4.1R also resulted in significantly decreased glucose uptake,lactate secretion and ATP production of the cells and increased pH value of the cell culture supernatant.qRT-PCR showed significantly decreased mRNA expressions of the key regulatory enzymes in glycolysis in 4.1R-/-HL-7702 cells.Compared with those in HL-7702 cells,the expression levels of AMPK and Raptor proteins were decreased while the expression levels of p-AMPK and p-Raptor proteins increased significantly in 4.1R-/-HL-7702 cells.Conclusion Deletion of protein 4.1R in HL-7702 cells results in reduced proliferative capacity,increased apoptosis and suppression of glycolysis,and this regulatory mechanism is closely related with the activation of the downstream AMPK-mTORC1 signaling pathway.

Objective:To evaluate the effect of goal-oriented online and offline mixed teaching method on the trainees of burn operating room.Methods:From June 2019 to June 2020, 42 trainees of burn operating room in our hospital were selected for the randomized parallel trial, and they were randomly divided into two groups, routine group and research group. The routine group adopted the conventional online and offline mixed teaching method, while the research group adopted the goal-oriented online and offline mixed teaching method. The internship time of both group lasted for 1 month. Results and excellent rates, self-confidence and burn surgery skills evaluation before and after the internship, and satisfaction with the internship mode were compared between the two groups. SPSS 19.0 was used for t test and chi-square test. Results:The results and excellent and good rates of theoretical examination and practical examination in the research group were higher than those of the routine group. The scores of self-confidence, choice of operation mode, innovation and optimization of operation, control of operation complications and treatment of intraoperative emergencies in the two groups after internship were higher than those before internship, and the above scores of research group were higher than those of the routine group after internship. The satisfaction scores of the students on enhancing self-confidence, and improving operational ability, learning initiative and learning efficiency in the research group were significantly higher than those in the routine group ( P < 0.05). Conclusion:Goal-oriented online and offline mixed teaching method on trainees in burn operating room can not only improve the examination results, enhance their confidence and burn surgery skills, but also achieve their satisfaction.