Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore the effect of down-regulation of long non-coding RNA (lncRNA) CTB-191K22.5 on the proliferation and invasion of colorectal cancer SW480 cells and the molecular mechanism.Methods:The TCGA database was used to analyze the expression differences of CTB-191K22.5 in colorectal cancer tissues and normal tissues. The CTB-191K22.5 inhibitor (Anti-CTB-191K22.5) and negative inhibitor (Control) were transfected into colorectal cancer SW480 cells, denoted as Observation group and Control group, real-time quantitative polymerase chain reaction (qRT) -PCR) was used to evaluate the inhibitory effect. MTT method and Transwell chamber method were used to evaluate the proliferation and invasion of SW480 cells. Western blot was used to evaluate the protein levels of PI 3K/AKT/mTOR signaling pathway in SW480 cells. The bioinformatics software starBase v2.0 was used to predict the target genes of CTB-191K22.5. qRT-PCR was used to evaluate the expression of CTB-191K22.5 target gene in SW480 cells. Measurement data were expressed as Mean±SD, and t-test was used for comparison between two groups. Results:Compared with normal tissues, the expression of CTB-191K22.5 in colorectal cancer tissues was significantly increased ( P<0.01). The expression of CTB-191K22.5 in SW480 cells of the Control group and Observation group were 6.60±0.85 and 1.08±0.21, respectively. The expression level of CTB-191K22.5 decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the SW480 cell proliferation ability of the Observation group decreased ( P<0.01). The invasion numbers of SW480 cells in the Control group and Observation group were (135.4 ± 16.29) and (42.24±14.59), respectively. The invasion ability of SW480 cells decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the expression levels of PI 3K/AKT/mTOR signaling pathway protein in SW480 cells in the Observation group decreased. miR-326 may be the target gene of CTB-191K22.5. Compared with the Control group, transfection with Anti-CTB-191K22.5 significantly increased the expression level of miR-326 in SW480 cells ( P<0.01). Conclusion:CTB-191K22.5 is highly expressed in colorectal cancer tissues, and down-regulation of CTB-191K22.5 may inhibit the proliferation and invasion of colorectal cancer SW480 cells by targeting miR-326.

Objective:To evaluate the post-marketing safety and immunogenicity of a 23-valent pneumococcal polysaccharide vaccine (PPV23).Methods:From September 2020 to June 2021, a clinical trial of single-dose PPV23 was conducted in people ≥3 years old in Centers for Disease Control and Prevention of Guizhou, Hunan and Fujian provinces. Blood samples were collects from the subjects before and 30 d after vaccination. ELISA was used to quantitatively detect IgG antibodies against capsular polysaccharides of 23 Streptococcus pneumoniae serotypes in serum samples. The adverse events (AEs) were monitored within 7 d after vaccination. Results:A total of 409 subjects were enrolled and included in safety analysis. Except for one with antibody level inversion, the other 408 participants were included in immunogenicity analysis. The levels of antibodies against the 23 Streptococcus pneumoniae serotypes were all increased after vaccination by an average of 4.24 folds. The two-fold growth rates of the antibodies ranged from 51.72% to 96.81% with a total two-fold growth rate of 78.59%. The overall rate of AEs was 27.14% (111/409). Local AEs were mainly pain, induration, redness and swollen. No serious adverse events related to vaccination occurred. Conclusions:This study preliminarily demonstrated the good immunogenicity and safety of PPV23 vaccine.

Objective:To explore the effect and mechanism of microRNA (miRNA)-4320 on the proliferation and migration of gastric cancer MGC803 cells.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-4320 in four gastric cancer cell lines(MGC803, HS-746T, SGC7901, BGC823) MGC803 cells were infected with recombinant lentivirus carrying miR-4320 interference fragments or blank lentivirus, and set as si-miR-4320 group and NC group. Thiazole blue colorimetry and Transwell small box experiment were used to detect the proliferation and migration of MGC803 cells after miR-4320 was down-regulated. The bioinformatics software RNAhybrid was used to predict the target gene of miR-4320. The targeting relationship between miR-4320 and target gene was verified by dual-luciferase reporter gene experiment. qRT-PCR and Western blot were used to detect the expression of miR-4320 target gene. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test or one-way ANOVA was used for comparison between groups. Results:The expression of miR-4320 in the four gastric cancer cell lines was significantly higher than that of normal gastric mucosal epithelial cells ( P<0.01). The expression of miR-4320 in MGC803 cells in the NC group and the si-miR-4320 group were 8.19±1.00 and 1.09±0.31, respectively. The miR-4320 interference fragment significantly reduced the expression of miR-4320 ( P<0.01). The absorbance of MGC803 cells in the si-miR-4320 group was significantly lower than that of the NC group ( P<0.05), and the migration ability was significantly lower than that of the NC group ( P<0.01). Suppressor of cytokine signaling1 ( SOCSI) is the target gene of miR-4320. Compared with the NC group, the SOCS1 gene expression in the si-miR-4320 group was significantly up-regulated ( P<0.01). Conclusions:The expression of miR-4320 is increased in gastric cancer cell lines. Down-regulating the expression of miR-4320 can inhibit the proliferation and migration of gastric cancer MGC803 cells by inducing the expression of SOCS1 gene.

Objective:To explore the effect of microRNA (miRNA)-3126-5p on the proliferation and migration of colorectal cancer cells by inhibiting the expression of LIM and SH3 protein 1 ( LASP1). Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-3126-5p in colorectal cancer cell lines (HT-29, HCT116, LoVo, SW480) and normal intestinal mucosal epithelial cells (HIEC). The cell line with the lowest expression level was selected as the experimental object. The experiment was divided into 2 groups: the negative control group (transfected with miR-NC) and the miR-3126-5p group (transfected with miR-3126-5p). Cells of each group were collected 48h after transfection. qRT-PCR method was used to detect the expression level of miR-3126-5p in each group. The MTS method and the scratch healing experiment were used to detect the proliferation level and migration ability of the cells in each group. The bioinformatics software microRNA.org and the dual-luciferase reporter gene experiment were used to predict and verify the target genes of miR-3126-5p, respectively. qRT-PCR and Western blot were used to detect the expression levels of target genes in each group of cells. Measurement data were expressed as mean±standard deviation ( Mean± SD), t test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:Compared with normal intestinal mucosal epithelial cells (HIEC), the expression level of colorectal cancer cell line miR-3126-5p was significantly reduced ( P<0.05), and the cell line with the lowest expression level was HCT116 cells ( P<0.01). The expression of miR-3126-5p in HCT116 cells in the negative control group and miR-3126-5p group were (1.05±0.16) and (7.91±1.26) respectively, and the difference was statistically significant ( t=5.40, P<0.01). Compared with the negative control group, the proliferation ability of HCT116 cells in the miR-3126-5p group was significantly reduced ( t=4.52, P<0.05), and the migration ability was significantly reduced ( P<0.01). microRNA.org shows that miR-3126-5p has complementary binding sites with LIM and SH3 protein 1 ( LASP1) gene mRNA. miR-3126-5p can target LASP1 mRNA ( P<0.01). Compared with the negative control group, the expression of LASP1 gene in HCT116 cells of the miR-3126-5p group was significantly reduced ( t=4.56, P<0.01). Conclusion:The expression of miR-3126-5p in colorectal cancer cell lines is low, and miR-3126-5p can reduce the proliferation and migration ability of colorectal cancer HCT116 cells by inhibiting the expression of the target gene LASP1.

Objective:To investigate the effect of long non-coding RNA (lncRNA) HAGLR on the proliferation and migration of gastric cancer cells by inhibiting the expression of microRNA (miRNA, miR)-93-5p.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of HAGLR in gastric cancer cell lines (HS-746T, BGC823, SGC7901, MGC803) and normal gastric mucosal epithelial cells (GES-1). Selected the cell line with the lowest HAGLR expression and transfected with the negative control plasmid (negative control group) or HAGLR-high-expression plasmid (HAGLR group) respectively. The MTS method and the scratch healing test were used to detect the proliferation and migration ability of the cells after transfection. The bioinformatics software miRcode database was used to predict the target gene of HAGLR, and the dual luciferase reporter gene experiment was used to verify the binding of HAGLR to the target gene. qRT-PCR was used to detect the expression of the target gene. Western blot was used to detect the expression of Hippo signaling pathway. The software SPSS 21.0 was used to conduct statistical analysis. The t test was used for comparison between two groups, and the one-way analysis of variance was used for comparison between multiple groups. Results:Compared with GES-1 cells, the expression level of HAGLR in gastric cancer cell lines was lower (all P<0.05), and the cell line with the lowest HAGLR expression was SGC7901 cells ( P<0.01). The HAGLR expression in SGC7901 cells in the HAGLR group and the negative control group were 1.03±0.13 and 9.75±1.10, respectively. The expression level of HAGLR in the negative control group was significantly lower than that in the HAGLR group ( t=7.87, P<0.01). Compared with the negative control group, the absorbance of SGC7901 cells in the HAGLR group was significantly reduced ( P<0.05), and the scratch healing rate was significantly reduced ( P<0.01). The miRcode database showd that HAGLR and miR-93-5p have complementary binding sites. The dual luciferase reporter gene experiment showed that HAGLR can complement miR-93-5p ( P<0.01). Compared with the negative control group, the expression of miR-93-5p in SGC7901 cells in the HAGLR group was significantly reduced ( P<0.01), and the expression of Hippo signaling pathway protein was significantly reduced (all P<0.01). Conclusions:HAGLR is low expressed in gastric cancer cell lines. HAGLR inhibits the proliferation and migration of gastric cancer SGC7901 cells by negatively regulating miR-93-5p.

Objective? To investigate the bacterial contamination of open suction tubing in intensive care units at different dwelling time and to explore the proper replacement frequency of suction tubing. Methods? The patients receiving tracheal intubation or tracheotomy in the Intensive Care Unit, Jinhua Municipal Central Hospital from November 2016 to February 2018 were selected by convenient sampling. Based on preliminary pilot experiment, totally 39 patients were selected. Samples from the internal opening of suction tubing connector at days 1, 3 and 7 were obtained for bacterial culture, and the patients' sputum was cultured as required. The type of contaminant bacteria at the internal opening of suction tubing connector was identified. The number of bacterial colony, number of bacterial genus and the consistency with patients' sputum culture at different time points were compared. Results? The bacteria at the internal opening of suction tubing connector were mainly pseudomonas aeruginosa, viridans streptococcus, baumanii and corynebacterium, which was slightly different from the findings of sputum culture. There was no statistically significant difference in the positive rate, number of bacterial colony, number of bacterial genus and the consistency with patients' sputum culture at the internal opening suction tubing connector at days 1, 3 and 7 (P＞ 0.05). Conclusions? The internal opening of open suction tubing is highly contaminated, but the bacterial contamination does not worsen over time. The suction tubing may be replaced as long as every 7 days.

Objective To investigate whether the male adulthood brain volumes changes are correlated to their prenatal earthquake stress experience or suffering depression after birth. Methods A total of 956 adult males born in Tangshan during the earthquake were visited, and finally 41 subjects were recruited with 10 subjects in the stress?depression group (experienced prenatal earthquake stress and developed depression after birth), 18 subjects in stress-health group (experienced prenatal earthquake stress but no history of mental disorders after birth), and 13 blank control. 3.0 T magnetic resonance imaging was conducted on all subjects to record T1?weighted imaging. The curved surface?based morphology approach was employed to analyze the images, and to assess the changes in cortical gray matter volume, cortical thickness, cortical surface area, and subcortical nuclear volume. Results (1) There was significant difference in the age and HAMD17 score among the three groups (P<0.01). (2) Compared with the blank control group and corrected by whole brain volume, the gray matter volume of the left superior frontal gyrus (t=3.889, P=0.031) was significantly smaller in the stress-health group; the cortical thickness of the left superior frontal gyrus (t=4.968, P=0.046), left superior parietal lobule, left postcentral gyrus (t=4.362, P=0.015), and the right superior parietal lobule (t=4.212, P=0.010) significantly reduced in the stress-health group;the gray matter volume of the left superior frontal gyrus (t=4.365, P=0.049), and the left superior and middle frontal gyrus (t=4.231, P=0.042) in the stress?depression group significantly reduced; the cortical thickness of left superior frontal gyrus (t=4.878, P=0.012), left superior parietal lobule, left postcentral gyrus (t=4.741, P=0.004), right superior parietal lobule (t=4.323, P=0.005), right superior parietal lobule and the right precuneus (t=4.523, P=0.013) significantly decreased in the stress?depression group. Furthermore, the cortical surface area of left superior temporal gyrus (t=4.386, P=0.027) in the stress?depression group is significantly smaller than the stress-health group. And the volume of the right amygdala (P=0.022) in the stress?depression group also decreased in comparison with the blank control group. Conclusions The prenatal earthquake stress and later developing depression after birth may result in the change of their adulthood brain volumes, probably with more contribution coming from the prenatal earthquake stress. The biggest change can be observed in the ones who have both factors as a synergistic effect.