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Objective:To investigate the effect of stress-induced protein Sestrin2 (SESN2) on necroptosis of mouse dendritic cell (DC) induced by lipopolysaccharide (LPS) combined with zVAD, a panaspartate-specific cysteine protease (caspase) inhibitor.Methods:The DC2.4 cell line derived from the bone marrow of mouse in the 3rd to 10th generations was cultured. The cells were stimulated with LPS for 0 hour, 6 hours, 12 hours, and 24 hours, and grouped according to the stimulation time points. Western blotting was performed to determine the protein expression of SESN2 in each group. Overexpression empty lentivirus (NC), SESN2 gene overexpression RNA sequence lentivirus (SESN2 LV-RNA), small interfering empty lentivirus (NS), and SESN2 gene small interfering RNA sequence lentivirus (SESN2 siRNA) were transfected into DC2.4 cells. After 72 hours of transfection, cell fluorescence expression was observed under the inverted fluorescence microscope. Cells in each transfection group were stimulated with LPS for 24 hours. The blank control groups were set up and cultured with phosphate buffered saline (PBS) for 24 hours. Western blotting was performed to measure SESN2 protein expression. In the same groups as above, cells were stimulated with LPS+zVAD for 24 hours. The blank control groups were set up and cultured with PBS for 24 hours. Western blotting was used to determine the expression of mixed lineage kinase domain-like protein (MLKL) and phosphorylated-MLKL (p-MLKL). The p-MLKL levels and the number of positive cells were observed using laser scanning confocal microscopy. The necroptotic cell ratios were assessed by both flow cytometry and Hoechst staining.Results:Compared to the LPS 0 hour group, the expression of SESN2 in the LPS 24 hours group showed a significant increase. Therefore, 24 hours was chosen as the subsequent stimulation time point. After successful lentivirus transduction and 24 hours of cultivation, the MLKL phosphorylation level in the SESN2 siRNA+LPS+zVAD group was significantly higher than that in the NS+LPS+zVAD group. The MLKL phosphorylation in the SESN2 LV-RNA+LPS+zVAD group was significantly lower than that in the NC+LPS+zVAD group. The MLKL phosphorylation levels in both the NS+LPS+zVAD group and the NC+LPS+zVAD group were obviously higher than those in the NS+PBS group and the NC+PBS group, respectively. Laser scanning confocal microscopy showed that the trends in quantity and fluorescence intensity of p-MLKL protein expressions were consistent with the above results. The results from flow cytometry analysis and Hoechst staining showed that the rates of cell necrotic apoptosis in SESN2 siRNA+LPS+zVAD group were significantly higher than those in NS+LPS+zVAD group [flow cytometry analysis: (30.800±1.153)% vs. (20.800±1.114)%, Hoechst staining: (75.267±0.451)% vs. (46.267±3.371)%, both P < 0.05], indicating that knocking down SESN2 further exacerbated the occurrence of necroptosis. The necrotic apoptosis rates in SESN2 LV-RNA+LPS+zVAD group were significantly lower than those in NC+LPS+zVAD group [flow cytometry analysis: (7.160±0.669)% vs. (19.240±2.322)%, Hoechst staining: (32.433±3.113)% vs. (48.567±4.128)%, both P < 0.05], indicating that overexpressing SESN2 reversed such response and markedly reduced the proportion of necroptotic cells compared to the corresponding empty vector group. Conclusion:SESN2 exhibits an inhibitory effect on necroptosis of DC in sepsis. Targeted SESN2 expression may regulate the process of DC-mediated immune response in sepsis.
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Objective:To investigate the role and significance of NUFIP-1-mediated ribophagy in apoptosis of dendritic cells (DCs) stimulated by lipopolysaccharide (LPS).Methods:Cultured mouse dendritic cell line DC2.4 were divided into the blank control group and LPS stimulation groups for 6, 12, 24, 48 and 72 h ( n=5). LPS subgroups were consistently cultured with 1 μg/mL LPS for the corresponding incubation time. Western blot was adopted to detect the expression levels of NUFIP-1 and autophagy-related proteins p62 and LC3B across groups. Laser scanning confocal microscopy (LSCM) was applied to detect the expression and cellular localization of NUFIP-1, with its co-localization with Lyso-tracker and LC3B, respectively. The silencing blank vector NS and silencing virus vector NUFIP-1 siRNA were transferred into DC2.4 ( n=3) and stimulated with 1 μg/mL LPS for 24 h. The apoptosis of DC2.4 was measured by flow cytometry analysis. The expression levels of apoptosis-related proteins were determined using Western blot, including cleaved caspase-3 and Bcl-2. One-way analysis of variance (ANOVA) was applied for comparison among multiple groups, and LSD-t method was used for subsequent pairwise comparison. A P<0.05 was considered statistically significant. Results:The results of Western blot showed that expression level of NUFIP-1 in DC2.4 revealed a trend of first increasing and subsequent decreasing upon LPS stimulation for different times (6, 12, 24, 48 and 72 h), and the expression level of NUFIP-1 in the LPS 24 h group was significantly higher than that in the blank control group [blank control group: (0.6786 ± 0.0820); LPS 24 h group: (1.4830 ± 0.1170); P<0.01]. Meanwhile, p62 expression in the LPS 24 h group was significantly lower than that in the blank control group [blank control group: (0.9087 ± 0.1235); LPS 24 h group: (0.3113 ± 0.5571); P<0.01]. Moreover, the conversion from LC3B-I to LC3B-II in the LPS 24 h group was significantly higher than that in the blank control group [blank control group: (0.5542 ± 0.1248); LPS 24 h group: (2.5310 ± 0.3119); P<0.01]. LSCM indicated that NUFIP-1 was predominantly located in the nucleus and perinuclear area in DC2.4. The fluorescence intensity of NUFIP-1 increased in a time-dependent manner from 6 h to 24 h after LPS stimulation, whereas a significant reduction could be observed at 48 h and 72 h after LPS stimulation. Meanwhile, the co-localization of NUFIP-1 with Lyso-tracker and LC3B was substantially reinforced in comparison with the blank control group. Transfection of NUFIP-1 siRNA through lentivirus transfection technology significantly down-regulated the expression level of NUFIP-1 in DC2.4, with statistical differences compared with the blank control group and empty vector group [blank control group: (0.6627 ± 0.1707); empty vector group: (0.6966 ± 0.1107); siRNA group: (0.1428 ± 0.0296); P<0.05]. Flow cytometry analysis revealed that the apoptotic rate of LPS-stimulated DC2.4 was significantly higher in the NUFIP-1 siRNA transfection group than that in the blank control group and empty vector group [blank control LPS 24 h group: (47.91% ± 1.006%); empty vector LPS 24 h group: (70.26% ± 1.011%); siRNA LPS 24 h group: (80.23% ± 2.094); P<0.01]. Western blot analysis of apoptosis-related protein further confirmed that the expression level of cleaved caspase-3 was significantly elevated in the NUFIP-1 siRNA transfection group compared to those of the blank control group and empty vector group under LPS challenge [blank control LPS 24 h group: (0.4748 ± 0.0876); empty vector LPS 24 h group: (0.2849 ± 0.0418); siRNA LPS 24 h group: (0.9733 ± 0.0525); P<0.01]. Likewise, expression of Bcl-2, an anti-apoptotic protein was significantly down-regulated in the siRNA LPS 24 h group [blank control LPS 24 h group: (0.7810 ± 0.0490); empty vector LPS 24 h group: (0.8292 ± 0.0729); siRNA LPS 24 h group: (0.3957 ± 0.0838); P<0.05]. Conclusions:NUFIP-1-mediated ribophagy is significantly activated in DC2.4 upon LPS stimulation, exerting an underlying protective effect on apoptosis.
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@#BACKGROUND: The aim of the present study is to describe the clinical correlates of hypotension and its associated outcomes in patients with acute organophosphorus poisoning (AOPP). METHODS: In this retrospective cohort study, we analyzed data pertaining to 871 patients with AOPP who were treated at two hospitals. Data from hypotensive and non-hypotensive patients were compared to identify clinical correlates of hypotension. We also evaluated the association between clinical parameters (including hypotension) and in-hospital mortality. RESULTS: The incidence of hypotension in AOPP patients was 16.4%. Hypotensive patients showed significantly higher in-hospital mortality (1.1% vs. 39.9%, P<0.001). Advanced age (odds ratio [OR] 1.25, 95% confidence interval [CI] 1.08-1.44), history of diabetes (OR 2.65, 95% CI 1.14-5.96), and increased white blood cell count (OR 1.06, 95% CI 1.03-1.09), plasma cholinesterase (OR 0.91, 95% CI 0.84-0.94), plasma albumin (OR 0.88, 95% CI 0.85-0.92), serum amylase (OR 1.01, 95% CI 1.01-1.02), and blood pH (OR 0.64, 95% CI 0.54-0.75) were significantly associated with hypotension. After adjusting for potential confounders, hypotension was associated with increased in-hospital mortality (hazard ratio 8.77-37.06, depending on the controlled variables). CONCLUSIONS: Hypotension is a common complication of AOPP and is associated with increased in-hospital mortality. Advanced age, history of diabetes, and changes in laboratory parameters were associated with hypotension in AOPP patients.
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Objective:Evaluate the application of Fourier transform infrared spectroscopy in the identification of homology of carbapenem-resistant Escherichia coli(CREC). Methods:A total of 26 carbapenem-resistant Escherichia coli strains were isolated from 9 provinces in China in 2018. The 900-1 200 cm -1 was selected as a spectral region for the Euclidean distance calculating and average linkage clustering between all isolates.The single nucleotide polymorphism (SNP) was analyzed by whole genome sequencing (WGS). Results:Twenty-six CREC strains were divided into 14 infrared spectros copy(IR) types by FTIR. The same IR type belonged to the same sequence type type.Compared with cluster analysis based on WGS, the consistency of FTIR cluster analysis was 92.3% (24/26).Conclusions:FTIR presented excellent performance in identification of homology of CREC.Besides, with the advantages of simple operation and rapid acquisition of results, FTIR may be a useful tool in clinical labs.
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BACKGROUND@#The mortality rate among patients with nasopharyngeal carcinoma (NPC) has improved significantly with the advent of chemoradiotherapy strategies. However, distant metastasis remains problematic. Tumor-specific reactivity in cancer patients has been detected exclusively in CD39+ T cells, particularly in CD39+CD103+ T cells. Circulating cancer-specific T cells are important for protecting against metastasis. This study aimed to evaluate the predictive value of circulating CD39+CD8+ T cells for metastasis in patients with NPC.@*METHODS@#We performed a cross-sectional, longitudinal study of 55 patients with newly diagnosed NPC of stage III-IVa. All patients were initially treated with standard combined chemoradiotherapy. Blood samples were obtained from 24 patients before and at 1 month and 6 months after treatment. T cell expression of CD39 and CD103, together with the markers of T cell exhaustion programmed death-1 (PD-1)/T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) and markers of cell differentiation CD27/CC-chemokine receptor 7/CD45RA, was examined by flow cytometry. The Wilcoxon rank-sum test analysis was used to analyze the differences between two groups. Kaplan-Meier analysis was used for analysis of progression-free survival (PFS).@*RESULTS@#The expression of circulating CD39+CD8+ and CD39+CD103+ CD8+ T cells was significantly higher in patients without distant metastasis (CD39+CD8+: 6.52% [1.24%, 12.58%] vs. 2.41% [0.58%, 5.31%], Z=-2.073, P=0.038 and CD39+CD103+CD8+: 0.72% [0.26%, 2.05%] vs. 0.26% [0.12%, 0.64%], Z=-2.313, P = 0.021). Most CD39+ T cells did not express PD-1 or Tim-3. Patients with high expression of CD39+CD103+CD8+ T cells had better PFS than patients with low expression (log rank value = 4.854, P = 0.028). CD39+CD8+ T cells were significantly elevated at 1-month post-treatment (10.02% [0.98%, 17.42%] vs. 5.91% [0.61%, 10.23%], Z = -2.943, P = 0.003). The percentage of advanced differentiated CD8+ T cells also increased at 1-month post-treatment compared with pre-treatment (33.10% [21.60%, 43.05%] vs. 21.00% [11.65%, 43.00%], Z = -2.155, P = 0.031). There was a significant correlation between elevated CD39+CD8+ T cells and increased effector memory T cells (intermediate stage: r = 0.469, P = 0.031; advanced stage: r = 0.508, P = 0.019).@*CONCLUSIONS@#CD39+CD8+ circulating T cells have preserved effector function, contributing to an improved prognosis and a reduced risk of metastasis among NPC patients. These cells may thus be a useful predictive marker for a better prognosis in patients with NPC.
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Humans , CD8-Positive T-Lymphocytes , Chemoradiotherapy , Cross-Sectional Studies , Longitudinal Studies , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/therapy , PrognosisABSTRACT
Objective:To explore the effects of Xuebijing injection and its component hydroxysafflor yellow A on coagulation and survival rates of septic rats.Methods:① Assessment of coagulation: 144 male Sprague-Dawley (SD) rats were divided into four groups by random number table: sham group, cecal ligation and puncture (CLP) induced sepsis model group (CLP group), CLP+Xuebijing group, and CLP+hydroxysafflor yellow A group, with 36 rats in each group. CLP was used for reproducing septic models. The cecum of the rats in the sham group was exposed by laparotomy and then returned to the abdominal cavity without CLP, while the other steps were the same as those in the CLP group. Rats in the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group were injected with Xuebijing (4 mL/kg, twice a day) or hydroxysafflor yellow A solution (0.378 g/L, 298 μg each time, twice a day) through caudal vein after operation. Levels of prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fib), and D-dimer in peripheral blood were measured by automatic coagulation analyzer at 6, 12, 24 hours after operation. The enzyme linked immunosorbent assay (ELISA) was applied to determine levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and soluble thrombomodulin (sTM) in peripheral blood. ② Analysis of survival rates: 120 rats were divided into four groups by random number table (the same groups with those in the section of assessment of coagulation), with 30 rats in each group. The Kaplan-Meier survival curve was plotted, and the cumulative survival rates were observed and recorded for 7 days after CLP surgery.Results:① Results of coagulation assessment: compared with the sham group, septic rats in the CLP group showed significant dysfunction in coagulation early, as evidenced by prolonged PT at 6 hours after CLP (s: 8.9±0.2 vs. 8.4±0.4, P < 0.01), and significantly increased levels of Fib, D-dimer, TFPI and sTM [Fib (g/L): 2.8±0.3 vs. 2.3±0.1, D-dimer (ng/L): 1.8±0.2 vs. 1.5±0.1, TFPI (ng/L): 131.1±10.9 vs. 102.8±10.5, sTM (μg/L): 27.2±1.2 vs. 19.8±2.9, all P < 0.01]. The coagulation dysfunction became more and more serious at 12 hours after operation, and further deteriorated with time. The use of both Xuebijing and hydroxysafflor yellow A revealed significant improvement in coagulation of septic rats at 6 hours, as shown by shortened PT (s: 8.3±0.2, 8.3±0.1 vs. 8.9±0.2, both P < 0.01), and decreased Fib, D-dimer, TFPI and sTM as compared with those in the CLP group [Fib (g/L): 2.3±0.1, 2.3±0.2 vs. 2.8±0.3; D-dimer (ng/L): 1.5±0.1, 1.5±0.2 vs. 1.8±0.2; TFPI (ng/L): 109.5±10.2, 91.5±5.0 vs. 131.1±10.9; sTM (μg/L): 22.3±1.5, 21.1±1.8 vs. 27.2±1.2; all P < 0.01]. However, there was no significant difference in coagulation function between the two intervention groups. ② Results of survival rates analysis: the rats in the sham group all survived 7 days after operation. The 7-day cumulative survival rate of the CLP group was only 36.67% (11/30). Compared with the CLP group, the cumulative survival rates were significantly increased in rats of the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group [66.67% (20/30), 66.67% (20/30) vs. 36.67% (11/30), both P < 0.05], but no significant difference was found between the CLP+Xuebijing group and CLP+hydroxysafflor yellow A group. Conclusion:Both Xuebijing and its component hydroxysafflor yellow A appear to be capable of alleviating coagulation disorders and improving survival rates of septic rats effectively, and the effects show no significant difference between them.
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Objective: To observe the inhibitory effect of astragalus polysaccharides (APS) on growth of human breast cancer MDA-MB-231 xenograft tumor in nude mice and its effect on the apoptosis of tumor cells, in order to study the effect of APS on growth and induction of apoptosis of triple negative breast cancer MDA-MB-231 and its possible molecular mechanism. Method: Human breast cancer cell MDA-MB-231 was inoculated into the right axillary subcutaneous of BALB/c-nu female nude mice to establish the transplanted tumor model of breast cancer. Eighteen nude mice were randomly divided into 3 groups:model group (saline per day), low-dose APS group (200 mg·kg-1 APS per day), and high-dose APS group (400 mg·kg-1 APS per day), with 6 rats in each group. The drug was administered by gavage (200 μL) daily for 21 days. In the experiment, the length and diameter of breast cancer transplanted tumor were measured every two days, and the tumor volume was recorded and calculated. At the end of the experiment, the changes of tumor mass and tumor volume of the low and high-dose APS groups and the model group were observed and compared, and the tumor inhibition rate was calculated. The cell morphology in tumor tissue was observed by hematoxylin-eosin (HE) staining, and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was used to verify the apoptosis of breast cancer tissues. The expressions of apoptosis-related proteins, such as B-cell lymphoma/leukemia-2 protein (Bcl-2), Bcl-2 associated X protein (Bax), Caspase in tumor tissues was detected by Western blot. Result: The tumor volume of breast cancer decreased in the low and high-dose APS groups, and the tumor inhibition rates were 37.9%and 57.57%, respectively, with statistically significant differences from the model group (PP0.01). HE of tumor tissue cells showed that APS led to obvious morphological changes, with apoptosis in the tissue cells. TUNEL staining showed that the apoptosis rate of tumor cells in APS intervention groups was higher than that in control group. Western blot showed that expression of Bcl-2 protein decreased(PPPPConclusion: APS can effectively inhibit the growth of MDA-MB-231 breast cancer xenografts in nude mice and induce apoptosis in human breast cancer MDA-MB-231 cells. The mechanism may be related to the effect of APS on expressions of apoptosis-related proteins Bcl-2, Bax, Caspase-9 and Caspase-7 in breast cancer cells.
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The Cd stress of Chrysanthemum indicum was treated by different concentrations of Cd Cl2 solution in the culture substrate. The content of Cd in different parts of Ch. indicum and the content of buddleoside and the total flavonoids in Ch. indicum were determined. The absorption characteristics of Cd elements in Ch. indicum were analyzed. And the influence of Cd elements on the quality of the herbs. The results showed that the application of soil Cd in the range of 0-100 mg·kg~(-1) had no significant effect on the biomass of Ch. indicum,and the root-shoot ratio showed a decreasing-increasing-decreasing trend. The content of Cd in different parts of Ch. indicum was significantly different,and the content of aboveground part was higher than that of underground part. The enrichment factors of Cd elements in different parts of Ch. indicum are different. The enrichment coefficient of aboveground parts is larger than that of underground parts. The whole parts and plants show an increase first and then decrease,and the overall enrichment factor is greater than1. The transfer coefficient of the aerial part/underground part of Ch. indicum showed a decreasing-increasing-decreasing-increasing trend with the increase of the amount of Cd applied in the soil,and the transfer coefficient was higher than 1. The contents of buddleoside and total flavonoids in Ch. indicum after Cd stress treatment were lower than the control,and the overall performance was lower and then increased,but it was still significantly lower than the control,indicating that Cd pollution directly led to the decrease of chemical quality of Ch. indicum.
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Cadmium , Chrysanthemum , Flavonoids , Soil , Soil PollutantsABSTRACT
@#Objective To compare the quality of life (QOL) of advanced Siewert type Ⅱ adenocarcinoma esophagogastric junction (AEG) patients treated by the total gastrectomy, traditional proximal gastrectomy or proximal gastrectomy with gastric tube reconstruction opertations, and to provide some clinical basis for the choice of surgical methods for AEG. Methods A total of 90 patients with Siewert type Ⅱ AEG were retrospectively collected from the Affiliated Hospital of North Sichuan Medical College. Patients were divided into 3 groups according to different surgical methods (n=30 in each group), a total gastrectomy group (23 males, 7 females, aged 47-79 years), a traditional proximal gastrectomy group (treated with the traditional proximal gastrectomy procedure, 19 males, 11 females, aged 44-80 years), and a narrow gastric tube group (treated with the proximal gastrectomy with gastric tube reconstruction procedure, 25 males, 5 females, aged 47-83 years). The Chinese version of Quality of Life Questionnaire Core-30 (QLQ-C30) and Quality of Life Questionnaire Oesophagogastric-25 (QLQ-OG25) designed by European Organization for Research and Treatment of Cancer (EORTC) were used to collect the patients’ information in the three groups about their QOL during the first six months and one year after the three procedures. Results There was no statistical difference in the clinical data among the three groups (all P>0.05). QOL during the first six months after the operations assessed by the QLQ-C30 questionnaire table showed that the narrow gastric tube group was significantly best in total QOL, physical function, fatigue, and emotional function among the three groups (all P<0.05). The total gastrectomy group was the worst in role function, dyspnea, fatigue and diarrhea among the three groups (all P<0.05). The traditional proximal gastrectomy group had a worse evaluation in lose of appetite than the other two groups (P<0.05). QOL during the first six months after the operations assessed by the QLQ-OG25 questionnaire table showed that the traditional proximal gastrectomy group had a significantly worse evaluation in palirrhea than the other two groups (both P<0.05). The total gastrectomy group had a significantly worse evaluation in anxiety than the other two groups (both P<0.05). QOL during the first year after the operations assessed by the QLQ-C30 questionnaire table showed that the narrow gastric tube group had a significantly highest evaluation in total QOL physical function and emotional function among the three groups (all P<0.05). The total gastrectomy group had a significantly worst evaluation in diarrhea among the three groups (P<0.05). QOL during the first year after operations assessed by QLQ-OG25 questionnaire table showed that the traditional proximal gastrectomy group had a significantly worse evaluation in palirrhea than the other two groups (all P<0.05). Conclusion The narrow tubular esophagogastric anastomosis is better than the total gastrectomy and the traditional proximal gastrectomy for the treatment of the advanced Siewert type Ⅱ adenocarcinoma of esophagogastric junction, so this operation is worth being recommended.
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BACKGROUND:The cyclic fatigue finite element method is more useful to study the fatigue life of the prosthesis in accordance with the actual situation of the oral cavity. OBJECTIVE:To study the all-ceramic fixed bridge fatigue life with different arch curvatures and materials. METHODS:According to the arch curvature, four sets of models (group Ⅰ, 0°; group Ⅱ, 30°; group Ⅲ, 60°; group Ⅳ, 90°) were set up on the basis of a digital model of all-ceramic fixed bridge. Fatigue life of each model was analyzed with different materials (In-Ceram Zirconia, Zenotec, LAVA). RESULTS AND CONCLUSION:The stress was mostly concentrated at the join of the all-ceramic fixed bridge. It was also the most part of the stress extremum, the shortest part of fatigue life and the most easily damaged part of the material. With the increase of arch curvature, the fatigue life of Group In-Ceram Zirconia dropped from 4.6 to 1.7 years, while both Group Zenotec and Group LAVA had the fatigue life of 19.6 years. The part of minimum life was the join of the fixed bridge. These findings indicate that the expected fatigue life of In-Ceram Zirconia all-ceramic fixed bridge restoration is relatively short, which may be an inappropriate repair material in anterior and posterior positions of canine teeth. All-ceramic materials, Zenotec and Lava, can meet the clinical life expectancy under different curvatures.
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Objective To evaluate the modified foruix-based technique with Guyton hook as an approach for the treatment of children with horizontal strabismus.Methods The clinical data of 128 patients (170 eyes) who underwent horizontal strabismus surgery between January 2014 and June 2017 were retrospectively reviewed,including 60 males and 68 females.The mean age was 1.5-17.0 (6.5 ± 1.6) years.All procedures under general anesthesia were performed using the modified fornix-based conjunctival incision with Guyton hook,and the follow-up period was 6 to 12 months.The clinical and cosmetic outcomes of strabismus surgery,the complications and interventions related to the incision were assessed.Results At 3 months after surgery,orthophoria with excellent cosmetics was achieved in 119 patients (93%) with the deviation ≤ ± 10△.Together 102 eyes (60%) had no intraoperative suture.Moreover,the incision was sutured with one stitch in 45 eye (26.5%),in 20 (11.8%) with 2 stitches,and in 3 (1.8%) with 3 stitches.During the follow up period,there was no severe intraoperative or postoperative complications.The swelling and redness of conjunctiva recovered quickly.Patients' discomfort was alleviated in a few days after the surgery.No oblivious scarring was found along the incision lines.Conclusion The modified fornix-based approach with Guyton hook is an effective and safe method for minimal incision surgery in children with horizontal strabis.
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Objective To analyze the safety and efficacy of autogenous femoral lateral iliotibial fascia(autologous fascia lata) in the treatment of female stress incontinence.Methods The clinical data of 7 female patients with stress incontinence admitted from January 2016 to June 2017 were retrospectively analyzed.The mean age was 58.2 years (range 45-72 years).The mean disease duration was 10.7 years (range 5-21 years).The mean Body mass index (BMI) was 24.1 kg/m2 (range 20.3-31.4 kg/m2).7 patients had severe subjective scores according to clinical symptoms.The average score of urinary incontinence questionnaire-simple form (ICIQ-SF) of international urinary incontinence advisory committee was 14.3 ± 1.1,the score of incontinence-quality of life (I-QOL) was 24.3 ± 4.8,respectively.During general anesthesia,the patient was placed in a half-recumbent position with the right leg straight down and the left leg bent over.The position of patella as well as the iliotibial band of the lateral femoral muscles were marked on the body surface.The iliotibial fascia of the lateral femoral muscle was exposed through a transverse incision 4-6 cm perpendicular to the iliotibial fascia of the lateral femoral muscle.The fascia of the lateral iliac tibial fascia of the lateral vastus with a width of 1.5 to 2.0 cm and a length of 12 to 14 cm was cut.The fascia was immersed in physiological saline and sutured with two 2-0 CT1PDS absorbable sutures or 2-0 vascular sutures at both ends of the fascia for 3 consecutive needles to form a sling.Then the urethral catheter was placed in the position of lithotomy,and then urethral suprapubic suspension was performed using autologous fascia through bilateral paravaginal incisions.The duration of surgery,intraoperative blood loss,intraoperative complications,postoperative catheter indwelling time,the length of hospital stay and postoperative complications were recorded.The situation of urinary incontinence as well as life quality before and after surgery were compared respectively.Results The operation was successfully performed.The mean operative time was 117.6 min(range 95-140 min).The mean intraoperative blood loss was 70.3 ml (range 50-90 ml).No complication was observed during the operation.The mean postoperative indwelling catheterization was 5.8 days (range 5-7 days).The mean postoperative hospital stay was 6.3 days (range 5-8 days).All 7 patients were cured after surgery,2 patients experienced urinary retention after removal of the catheter.The mean follow-up time was 18.2 months(range 13-24 months).No urinary incontinence or complications was observed postoperatively in all cases.Patient ICIQ-SF urinary incontinence questionnaire summary score and quality of life score of I-QOL questionnaire of 1 year post operation were 0.6 ± 0.5 and 96.1 ± 4.3,which were significantly improved compared with that before surgery(P < 0.01).Conclusion It is safe to use autologous femoral lateral muscle iliotibial fascia in the middle segment of urethra suspension for the treatment of female stress incontinence,and the curative effect is affirmative through one-year observation.
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Objective To investigate the potential role of Sestrin2 (SESN2) in regulating the apoptosis of dendritic cells (DCs) induced by high mobility group box-1 protein (HMGB1).Methods DCs (the murine DC cell line DC2.4) were cultured with or without HMGB1 stimulation (cultured with 10ng/ml HMGB1 for 8,24 and 48 hours,or cultured with HMGB1 for 48 hours at different concentrations of 1,10 and 100ng/ml,respectively,n=4).The protein level of SESN2,cleaved-caspase-3 and Bcl-2 were analyzed with Western blotting.Localization of SESN2 in cells was observed under confocal laser microscope (LSCM).Cell apoptosis was analyzed with flow cytometry.In addition,DC2.4 cells were transfected with lentivirus containing SESN2 LV-RNA,SESN2 siRNA sequence expressing plasmids or blank vector (NC,NS,n=4).These cells were then stimulated with HMGB1 (100ng/ml)for 48 hours,and the apoptosis was accessed as mentioned above.Results Compared with the control group,the expression of SESN2 was obviously up-regulated after HMGB1 (10ng/ml) stimulation for 24 and 48 hours (P<0.05).In a dose-dependent response,the expression of SESN2 was markedly enhanced in treatment with 1,10,100ng/ml HMGB1 for 48 hours (P<0.05).Compared with the control group (7.35% ± 1.33%),the percentage of apoptosis was significantly increased with 10,100ng/ml HMGB1 for 48 hours [(17.02% ± 4.85%,17.48% ± 4.04%,respectively,P<0.05 or P<0.01].After transfection,compared with blank vector group,the apoptosis of SESN2 siRNA group obviously elevated [(65.96% ± 2.50%) vs.(50.01% ± 2.07%),P<0.05],and cleaved-caspase-3 expression significantly increased while Bcl-2 expression obviously decreased.In SESN2 LV-RNA group,the apoptosis significantly decreased [(35.57% ± 1.69%) vs.(49.04% ± 4.87%),P<0.05],and cleaved-caspase-3 expression decreased and Bcl-2 expression obviously increased compared with blank vector group (P<0.05).Conclusion SESN2 has a protective effect against HMGB 1 induced apoptosis of D C2.4 cells.
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Diabetic retinopathy (DR) is one of the most significant microvascular complications of diabetes,and its incidence increases with the prolongation of DM;meanwhile,it can lead to severe vision loss.Recently,the pathogenesis of DR has been one of the focuses both at home and abroad,but the basic mechanisms are not fully elucidated.More and more scholars believe that DR may be a chronic and low degree of inflammatory response.Moreover,it is well established that microglia is a monocyte-macrophage,with antigen-presenting function.Especially in the retina,as microglia is activated,followed by the release of a large number of inflammatory factors,microglia involves in DR inflammatory response.Understanding the activation mechanism of microglia can provide a new perspective for the prevention and treatment of DR.Therefore,recent advances in activation of microglia in diabetic retinopathy and its intervention mechanism will be reviewed in the article.
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Objective This study aims to understand the characteristics of the time sequence of ICR mouse first mandibular molar tooth germ development through dynamic observation.Methods Tooth germ of Embryos (E11.5,E12.5,E13.5,E14.5,E15.5,E16.5,E17.5 and E18.5) and postnatal (PN1,PN2) mice were obtained.The heads (E11.5-E15.5) and mandibles (E16.5-PN2) of mice were dissected,fixed and embedded for serial sections and HE staining.All the results were assessed under light microscopy.Results The tooth germ underwent various development stages including the bud,cap and bell stages.Mouse odontogenesis was initiated at E11.5.Proliferation of oral epithelium formed the bud stage at E13.5.Then the cap stage was observed at E14.5-E15.5 and the bell stage was appeared beginning from E16.5.The pre-dentin was observed at PN1,as well as the dentin at PN2.Conclusions Establishing the regular development pattern of the first mandibular molar of ICR mice will provide a reliable basis for the future use in the specific tooth germ developmental research.
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Objective To investigate the potential effect of glucagon-like peptide-1 (GLP-1) analogue exendin-4 (Ex-4) on immune function of T lymphocytes via neuroendocrine modulation mechanism in mice following severe burns.Methods Male BALB/C mice were randomly (ramdam number) divided into thermal injury group (n =50) and sham-thermal group (n =30).The thermal injury model was made by exposing the back skin of 15% total body surface area (TBSA) to 95 ℃ water for 7 seconds,while in sham-thermal model the mice were immersed in 37 ℃ water instead.The expression of GLP-1 receptor (GLP-1 R) was determined in sorted CM + T cells from normal mice by immunofluorescence method.In ex vivo experiment,the mice were sacrificed at 24 h post-bum,then the mononuclear cells (MNC) from spleen were separated from both groups and cultured in RMPI 1640 with 10% FCS (fetal calf serum) in presence of ConA (concanavalin A,5 μg/mL).Cells were pretreated with catecholamine receptor antagonist propranolol (prop) for 1 hour,followed by consecutive dose of Ex-4 for another 48 h.In in vivo experiment,prop (30 mg/kg) was i.p.injected 30 minutes before thermal injury,then Ex-4 (2.4 nmol/kg) was injected i.p.immediately after scalding.Mice were sacrificed at 6 h and 24 h after thermal injury,then the serum and the spleens were collected.Results GLP-1R was expressed on splenic CD4 + T cells from normal micc.Ex-4 exerted no marked effect on the functions of T cells in terms of proliferation and IL-2 secretion at all doses examined ex vivo,which was not affected by pretreatment with prop.In vivo,T cell functions were suppressed by Ex-4 in thermal mice (P < 0.05),but was restored by pretreatment with prop.Regardless of ex vivo or in vivo,Ex-4 could induce T cells switched to Th2 response (P < 0.05).Moreover,the Th2 switch by Ex-4 was greatly potentiated by prop intervention in thermal mice in vivo other than ex vivo.Norepinephrine level was increased and epinephrine was decreased by Ex-4 in thermal mice.Both norepinephrine and epinephrine levels were obviously enhanced by pretreatment with prop.Conclusions Ex-4 can inhibit the proliferation and IL-2 secretion of splenic T lymphocytes through the sympathetic nervous system,however,it might induce Th2 switch from Th cells by acting directly on GLP-1R.
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Objective To explore the MRI features about the annular ligament injuries in pediatric patients. Methods The appearance of MRI about annular ligament injuries in 15 pediatric patients diagnosed clinically were retrospectively analyzed. All patients (aged 4 years 8 months to 12 years 11 months) were scanned by MRI in 2-4 days after injury. Patients were divided into three groups according to the annular ligament injuries severity, which were simple annular ligament injuries, annular ligament injuries combined dislocation, annular ligament ruptures combined dislocation. Results Simple annular ligament injuries were showed in 2 cases, which appeared high signal but continuous configuration. Annular ligament injuries combined dislocation were showed 9 cases. Bald head sign, thickening and continuous ligment were found on axial image, together with compression of the radial joint space on sagittal and coronal view. Annular ligament ruptures combined dislocation were showed in 4 cases. The ligment showed"bald head sign", tortuosity and uneven thickening on axial view and compression of the radial joint space on sagittal and coronal view. Combining with the other tissue injuries were detected in 15 cases. Conclusion The annular ligament injuries show some MRI characteristics. Bald head sign is specific in the annular ligament injuries of pediatric patients.
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Objective To explore the MRI features about the annular ligament injuries in pediatric patients. Methods The appearance of MRI about annular ligament injuries in 15 pediatric patients diagnosed clinically were retrospectively analyzed. All patients (aged 4 years 8 months to 12 years 11 months) were scanned by MRI in 2-4 days after injury. Patients were divided into three groups according to the annular ligament injuries severity, which were simple annular ligament injuries, annular ligament injuries combined dislocation, annular ligament ruptures combined dislocation. Results Simple annular ligament injuries were showed in 2 cases, which appeared high signal but continuous configuration. Annular ligament injuries combined dislocation were showed 9 cases. Bald head sign, thickening and continuous ligment were found on axial image, together with compression of the radial joint space on sagittal and coronal view. Annular ligament ruptures combined dislocation were showed in 4 cases. The ligment showed"bald head sign", tortuosity and uneven thickening on axial view and compression of the radial joint space on sagittal and coronal view. Combining with the other tissue injuries were detected in 15 cases. Conclusion The annular ligament injuries show some MRI characteristics. Bald head sign is specific in the annular ligament injuries of pediatric patients.
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<p><b>OBJECTIVE</b>To investigate the changes in plasma gelsolin (pGSN) levels in severe burn patients with sepsis, and to evaluate the prognosis of patients when combined with other related clinical indexes.</p><p><b>METHODS</b>Sixty-five severe burn patients with sepsis hospitalized from June 2013 to June 2015 conforming to the study criteria were divided into death group (n=24) and survival group (n=41) according to the clinical outcome on post sepsis diagnosis day (PSD) 28. The pGSN levels of patients were determined on PSD 1, 3, 7, and 14 with double antibody sandwich enzyme-linked immunosorbent assay. The serum level of C-reactive protein (CRP), serum level of procalcitonin, lactate level of arterial blood, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and Sequential Organ Failure Assessment (SOFA) score were determined or recorded on PSD 1. Data were processed with repeated measurement analysis of variance, t test, and chi-square test. On PSD 1, the pGSN level, serum level of CRP, serum level of procalcitonin, lactate level of arterial blood, APACHE II score, and SOFA score of 65 patients were collected to screen the independent risk factors related to death with single factor and multi-factor Logistic regression analysis. Receiver operating characteristic (ROC) curves of the independent risk factors related to death were plotted to evaluate the predictive power for death in 65 patients.</p><p><b>RESULTS</b>(1) The pGSN levels of patients in death group on PSD 1, 3, 7, and 14 were respectively (146±44), (85±24), (28±7), and (19±4) mg/L, obviously lower than those in survival group [(287±82), (179±51), (196±56), and (249±67) mg/L, with t values from 1.735 to 4.304, P<0.05 or P<0.01]. (2) The serum level of CRP, serum level of procalcitonin, lactate level of arterial blood, APACHE II score, and SOFA score of patients in death group on PSD 1 were respectively (56±7) mg/L, (12.54±0.82) μg/L, (2.74±0.27) mmol/L, (24.3±2.4) points, and (11.43±0.57) points, significantly higher than those in survival group [(35±4) mg/L, (2.38±0.16) μg/L, (1.83±0.12) mmol/L, (15.0±1.5) points, and (7.22±0.23) points, with t values from 1.902 to 3.883, P<0.05 or P<0.01]. (3) Multi-factor Logistic regression analysis showed that the pGSN level (odds ratio: 6.83, 95% confidence interval: 4.33-10.25, P<0.01) and APACHE II score (odds ratio: 5.27, 95% confidence interval: 2.28-9.16, P<0.01) were the independent risk factors related to death in 65 patients on PSD 1. (4) The total areas under the ROC curves of pGSN level and APACHE II score for predicting death of 65 patients on PSD 1 were respectively 0.89 and 0.86, and 142 mg/L and 21 points were respectively chosen as the optimal threshold values, with sensitivity of 87% and 83% and specificity of 86% and 89%.</p><p><b>CONCLUSIONS</b>For severe burn patients with sepsis, lowering of pGSN level and elevation of APACHE II score are obviously correlated with increase in case fatality rates. Monitoring the dynamic changes in pGSN level and APACHE II score during the early stage may be useful to predict the prognosis of severe burn patients with sepsis.</p>
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Humans , Burns , C-Reactive Protein , Calcitonin , Blood , Calcitonin Gene-Related Peptide , Enzyme-Linked Immunosorbent Assay , Gelsolin , Blood , Hospitalization , Organ Dysfunction Scores , Prognosis , Protein Precursors , Blood , ROC Curve , Regression Analysis , Risk Factors , Sepsis , Blood , Diagnosis , Severity of Illness IndexABSTRACT
Objective To investigate changes in high mobility group box-1 protein ( HMGB1 ) level in brain tissues with severe sepsis, and the relationship between HMGB1 and septic brain injury.Methods Forty wild C57BL/6 mice were randomly ( random number) divided into 4 groups: sham group, sepsis group, cerebroventricular injection control group, and sepsis with BoxA ( HMGB1 inhibitor) cerebroventricular injection group.Septic model was reproduced by cecal ligation and puncture, and the cerebroventricular catheterization model was established by motorized mice brain stereotaxic instruments.After septic challenge, 1 μg BoxA was injected into the ventricle of brain via cerebroventricular catheter immediately.Mice were sacrificed and brains were harvested at 24 h after sepsis, and hippocampus tissue was separated immediately.Expressions of brain HMGB1 and caspase-3 changed in apoptotic neurons and brain injury were determined by brain tissue immunofluorescence, Western blotting, TUNEL and HE staining respectively.One-way analysis of variance ( ANOVA) for analyzing inter-group differences, student t test for comparing difference between two groups . Results (1) HMGB1 expression in hippocampus was significantly enhanced in the septic group compared to the sham group [ (22.74 ±9.29) vs.4.57 ±2.18, P<0.01].(2) Compared to the sham group, neuronal apoptosis [ (35 ±9.17) vs.(1.67 ±1.53) , P<0.01) and caspase-3 expressions [ (16.79 ±8.17) vs.( 3.39 ±2.09), P<0.05] were significantly increased in hippocampus with aggravated brain injury in the septic group.(3) Cerebroventricular injection of BoxA significantly inhibited HMGB1 in hippocampus [ (2.66 ± 2.06) vs.( 22.74 ±9.29), P<0.01];(4) Cerebroventricular injection of BoxA obviously alleviated acute brain injury, and decreased neuronal apoptosis [ ( 12 ±4.36 ) vs.( 35 ±9.17 ) , P <0.01 ] as well as caspase-3 activity [ (4.10 ±2.11) vs.(16.80 ±8.17), P<0.05].Conclusions The elevated expression of brain HMGB1 is closely related to pathogenesis and development of septic brain injury, and treatment with antagonist towards brain HMGB1 can markedly attenuate acute brain injury following severe sepsis.