We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.

Objective: To observe the effects of ω-3 polyunsaturated fatty acids (PUFA) on damage of intestinal mucosa of rats with severe burn in early stage and to explore the mechanism. Methods: One hundred and twenty SD rats were divided into sham injury group, pure burn group, and ω-3 PUFA group according to the random number table, with 40 rats in each group. Rats in sham injury group were sham injured, while rats in pure burn group and ω-3 PUFA group were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burn) on the back. Rats in sham injury group and pure burn group were injected with normal saline solution (1 mL/kg) by tail vein, while rats in ω-3 PUFA group were injected with ω-3 PUFA solution (1 mL/kg) by the same way at 5 minutes post injury. At post injury hour (PIH) 3, 6, 12, 24, and 48, abdominal aorta blood and intestinal mucosa were collected from 8 rats in each group, respectively. Serum content of diamine oxidase (DAO) was detected by spectrophotography. Serum content of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) was determined by enzyme-linked immunosorbent assay. Protein expression of NF-κB-p65 in intestinal mucosa was determined by Western blotting. Data were processed with analysis of variance of factorial design, one-way analysis of variance, chi-square test, LSD test, and Bonferroni correction. Results: (1) At all time points post injury, serum content of DAO of rats in pure burn group and ω-3 PUFA group was significantly higher than that in sham injury group (with P values below 0.01), and serum content of DAO of rats in ω-3 PUFA group was significantly lower than that in pure burn group (with P values below 0.01). (2) At all time points post injury, serum content of TNF-α and IL-6 of rats in pure burn group and ω-3 PUFA group was significantly higher than that in sham injury group (with P values below 0.01), and serum content of TNF-α and IL-6 of rats in ω-3 PUFA group was obviously lower than that in pure burn group (with P values below 0.01). (3) At all time points post injury, protein expressions of NF-κB-p65 in intestinal mucosa of rats in pure burn group and ω-3 PUFA group were significantly higher than those in sham injury group (with P values below 0.01). At PIH 3, 6, 12, 24, and 48, protein expressions of NF-κB-p65 in intestinal mucosa of rats in ω-3 PUFA group were 1.398±0.016, 1.999±0.948, 2.803±0.065, 1.739±0.602, and 1.484±0.645, obviously lower than 2.096±0.113, 3.402±0.189, 4.183±0.558, 3.618±0.408, and 2.614±0.775 in pure burn group (with P values below 0.01). Conclusions The ω-3 PUFA may alleviate intestinal mucosa injury of rats with severe burn in early stage through reducing protein expression of NF-κB-p65 of intestinal mucosa, serum content of DAO, TNF-α, and IL-6, and inhibiting inflammatory response.

Objective To investigate the value of contrast-enhanced ultrasound(CEUS)combined with shear wave elastography(SWE)in the diagnosis of liver tumors.Methods This study was conducted according to the PRISMA guideline,with a PROSPERO registration number of CRD42023491288.PubMed,Embase,the Cochrane Library,CNKI,VIP,and Wanfang Data were searched for articles on CEUS combined with SWE in the diagnosis of liver tumors published from January 2000 to October 2023,and a total of 12 articles were included,with 1 328 patients in total.The QUADAS-2 tool was used to assess the quality of the articles included.Stata 15.0 software was used to calculate pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio,and heterogeneity.The summary receiver operating characteristic(SROC)curve was plotted,and the area under the SROC curve(AUC)was calculated.Results There were 1 457 lesions for the patients included,among whom there were 764 malignant lesions and 693 benign lesions,with a positive rate of 52.44%and a negative rate of 47.56%.Calculations obtained a pooled sensitivity of 0.94(95%confidence interval[CI]:0.91—0.96),a specificity of 0.92(95%CI:0.87—0.95),a positive likelihood ratio of 12.00(95%CI:7.40—19.40),a negative likelihood ratio of 0.06(95%CI:0.04—0.10),and a diagnostic odds ratio of 191(95%CI:87—417).The tests for heterogeneity showed Q=54.78,df=11.00,P<0.001,and I2=79.92%(95%CI:69.18%—90.66%),with an AUC of 0.98.Conclusion CEUS combined with SWE has a relatively high diagnostic value for benign and malignant liver tumors and thus holds promise for clinical application.

Objective To construct a mutant strain of Nocardia farcinica ( N. farcinica ) IFM10152 with mammalian cell entry 4A gene (mce4A) deletion and to analyze the function of that gene dur-ing infection. -ethods The mutant strain of N. farcinica was constructed through in-frame deletion without antibiotic labeling and verified by PCR and sequencing analysis. To analyze the function of mce4A gene in the interaction between N. farcinica and host cells, in vitro growth experiment, macrophage killing experi-ment using THP-1 ( a human leukemia mononuclear cell line) as the model and adhesion and invasion exper-iments using HeLa cells ( cervical cancer epithelial cells) were carried out. Results The mutant strain with mce4A gene deletion was successfully constructed and named △mce4A. No significant difference in growth rate was observed between the mutant and the wild-type strains. After knocking out the mce4A gene, the ability of N. farcinica to resist macrophage killing was obviously weakened as well as its ability to adhere and invade. Conclusions The mutant strain of N. farcinica with mce4A gene deletion was successfully construc-ted. The mce4A gene might play an important role in the adhesion and invasion of N. farcinica to host cells and its survival in macrophages.

Objective To investigate the effects of high glucose and lysophosphatidylcholine (LPC) on the immune function of in vitro cultured macrophages during Nocardia farcinica infection. Meth-ods RAW264.7 macrophages were cultured in vitro under different conditions as follows: routine culture (control group),50 mmol/L glucose (high glucose group),10 mg/L LPC(LPC groupⅠ),25 mg/L LPC (LPC groupⅡ) and 50 mmol/L glucose+25 mg/L LPC(high glucose and LPC group). The activity of mac-rophages in each group was tested after 6,12,24 and 36 h of culture. After 24 h of culture, macrophages were collected from every group and co-cultured with Nocardia farcinica. Dynamic phagocytosis rates were detected at 1,2,3,4,5 and 6 h after co-culture. Toxic effects of Nocardia farcinica on macrophages and concentrations of IL-10 and TNF-α were measured at 1,3 and 6 h after co-culture. Results Macrophages in all four experimental groups showed decreased activity as compared with those in the control group (P<0.01). Phagocytosis of Nocardia farcinica by macrophages was also reduced by high glucose and LPC. Phagocytosis rates of high glucose group and LPC groupⅡ at 1 and 2 h,LPC groupⅠat 1,2 and 3 h,and high glucose and LPC group at 1,2,3 and 4 h after co-culture were significantly lower than that of the con-trol group (P<0.05 or P<0.01). Compared with the control group, significantly reduced toxic effects on macrophages caused by Nocardia farcinica was observed in the experimental groups (P<0.05 or P<0.01). Compared with the control group,LPC groupsⅠand Ⅱ and high glucose and LPC group had decreased se-cretion of IL-10 at 3 h,and high glucose group and LPC groupⅠhad decreased secretion of TNF-α at 1 h(P<0.05). Conclusion Culture macrophages under the conditions of high glucose and LPC would reduce their activity and impair their ability to phagocytose Nocardia farcinica. Moreover, high glucose and LPC might have impacts on the toxic effects of Nocardia farcinica on macrophages and the secretion of IL-10 and TNF-α.

Objective:To explore the influence of parental compliance on the treatment of hypertrophic scars in burn children.Methods:A retrospective cohort study method was used. From June 2014 to June 2019, 49 children with post-burn hypertrophic scars who met the inclusion criteria and visited the outpatient department of the Department of Burns of the First Affiliated Hospital of Anhui Medical University were included in this study. In the follow-up of 9 months, according to the registration form and the results of the compliance questionnaire for parents, the children were divided into good compliance group (34 cases, 21 males and 13 females, aged 2.0 (2.0, 3.5) years) and poor compliance group (15 cases, 6 males and 9 females, aged 3.0 (2.0, 4.0) years). At the first attendance and in the follow-up of 3, 6, and 9 months, the scar scores of children in good compliance group were evaluated by Vancouver Scar Scale (VSS). At the first attendance and in the follow-up of 9 months, the scar scores of children in poor compliance group were evaluated by VSS. At the first attendance and in the follow-up of 9 months, the scar pruritus scores of children in the 2 groups were evaluated by Verbal Rating Score (VRS). Data was statistically analyzed with chi-square test, Wilcoxon rank sum test, Mann-Whitney U test, independent sample t test, and paired sample t test. Results:At the first attendance, the color, vascular distribution, softness, and thickness scores, and total score in VSS scoring of scars of children in the two groups were similar ( Z=0.834, 0.026, 0.837, 0.076, 1.074, P>0.05). In the follow-up of 9 months, the softness and thickness scores, and total score in VSS scoring of scars of children in good compliance group were significantly lower than those in poor compliance group ( Z=5.518, 4.732, 5.042, P<0.01). Compared with those in the first attendance, the color, vascular distribution, softness, and thickness scores, and total score in VSS scoring of scars of children in good compliance group were significantly decreased in the follow-up of 9 months ( Z=5.241, 5.273, 5.214, 5.245, 3.451, P<0.01); the color and vascular distribution scores, and total score in VSS scoring of scars of children in poor compliance group were significantly decreased in the follow-up of 9 months ( Z=3.606, 3.542, 3.448, P<0.01). At the first attendance, the VRS score of scar pruritus of children in good compliance group was 6.00 (5.00, 6.25) points, which was similar to (5.47±1.69) points in poor compliance group ( Z=0.607, P>0.05). In the follow-up of 9 months, the VRS score of scar pruritus of children in good compliance group was 1.00 (1.00, 1.25) points, which was significantly lower than (3.27±1.71) points in poor compliance group ( Z=2.606, P<0.01). Compared with those in the first attendance, the VRS score of scar pruritus of children in good compliance group was significantly decreased in the follow-up of 9 months ( Z=4.002, P<0.01), while there was no obvious change in poor compliance group in the follow-up of 9 months ( t=3.550, P>0.05). Conclusions:Under the same treatment plan, good parental compliance has a positive effect on the treatment of hypertrophic scars in burn children decreasing the degree of scar hyperplasia and pruritus.

OBJECTIVETo study characteristics of energy metabolism in the skeletal muscle of rats with postoperative fatigue syndrome (POFS) and the interventional effect of ginsenoside Rb1.

METHODWe chose resection of 70% of the "middle" small intestine as the rat model for POFS. Ninety-six adult male SPF SD rats were randomly divided into the control group, the model group, and the ginsenoside Rb1-treated group by body weight. And then, each group was further randomly divided into four subgroups, according to different postoperative investigated time points, such as postoperative day 1, postoperative day 3, postoperative day 7 and postoperative day 10. So the animals were divided into twelve subgroups (n = 8 in each subgroup). Rats of the control group and the model group were injected intraperitoneally with saline at the dose of 10 mL x kg(-1) one hour before the operation and once a day during the postoperative days. Rats of the ginsenoside Rb1-treated group were administered 10 mg x kg(-1) ginsenoside Rb1 by the same method. The skeletal muscles were sampled on postoperative day 1, 3, 7 and 10. The contents of ATP, ADP, AMP in skeletal muscles were determined by HPLC, and the activities of Na(+)-K(+)-ATPase and Ca(2+)-ATPase were investigated by colorimetry.

RESULTCompared with the control group, the content of ATP in skeletal muscle of rats of the model group decreased significantly on postoperative day 3 (P < 0.05), while the content of ADP significantly increased on postoperative day 7 and 10 (P < 0.05). The activity of Na(+)-K(+)-AT-Pase decreased on postoperative day 3 and 7 (P < 0.05), and the activity of Ca(2+)-ATPase decreased on postoperative day 7. After supplement of ginsenoside Rb1, on the investigated time points, all the negative changes of the indicators discovered above were significantly adjusted (P < 0.05) in rats of the ginsenoside Rb1-treated group, while no significant differences were investigated.

CONCLUSIONDuring a certain period of postoperative time, the activity of energy metabolism is depressed in the skeletal muscle of rats with POFS, but it can be improved by supplement of ginsenoside Rb1.

Animals ; Calcium-Transporting ATPases ; physiology ; Energy Metabolism ; drug effects ; Fatigue ; drug therapy ; metabolism ; Ginsenosides ; pharmacology ; therapeutic use ; Male ; Muscle, Skeletal ; metabolism ; Postoperative Complications ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; physiology ; Syndrome

Objective To explore the mechanism of hypertonic salt solution (HS) alleviates lung injury of rats at the early stage of severe scald.Methods Thirty-two female Sprague-Dawley (SD) rats were randomly assigned to sham group, lactated Ringer solution (LR) group, HS200 group (200 mmol/L HS group, 1 L 200 mmol/L HS contained 955 mL LR and 45 mL 10％ NaCl) and HS400 group (400 mmol/L HS group, 1 L 400 mmol/L HS contained 846 mL LR and 154 mL 10％ NaCl), with 8 rats in each group. A 30％ total body surface area (TBSA)Ⅲ degree scalded model was reproduced by scalded on the back with 98℃ boiling water for 12 seconds, whereas those in the sham group were exposed to 37 ℃ water without liquid resuscitation. Rats in the three drug intervention groups were resuscitated with LR, 200 mmol/L HS and 400 mmol/L HS by caudal vein according to the Parkland formula, respectively. All rats were sacrificed at 8 hours after scald injury to harvest abdominal aorta blood and lung tissues. Interleukins (IL-6, IL-10 and IL-17) in serum were determined by enzyme-linked immunosorbent assay (ELISA). Samples from the lung tissue were used to measure malondialdehyde (MDA) and superoxide dismutase (SOD) levels by ultraviolet spectrophotometer. Expressions of p38 mitogen-activated protein kinase (p38MAPK) and extracellular regulated protein kinase 1/2 (ERK1/2) in the lung were determined by Western Blot. The lung tissue was stained with hematoxylin and eosin (HE), and the pathological changes were observed with a light microscope.Results Compared with the sham group, the lung tissues in the LR group were damage obviously, which accompanied with more inflammatory cell infiltration, cell edema and pulmonary septum thickening, and the levels of IL-6, IL-10, IL-17 in serum and MDA content, the phosphorylation of p38MAPK and ERK1/2 in lung tissues were increased whereas the activity of SOD was decreased. Compared with the LR group, the lung injury was significantly alleviated, the levels of IL-6, IL-17 in serum and MDA content and the phosphorylation of p38MAPK and ERK1/2 were decreased, and the levels of IL-10 and SOD were increased in both HS groups with a dose-dependent manner. There were significant difference in above parameters between HS400 group and LR group [serum IL-6 (ng/L): 3.76±0.12 vs. 6.72±0.90, serum IL-10 (ng/L): 33.76±3.71 vs. 16.77±3.19, serum IL-17 (ng/L): 103.52±2.78 vs. 124.96±4.96, lung MDA (nmol/mg): 5.59±0.24 vs. 7.09±0.39, lung SOD (U/mg):226.7±3.9 vs. 172.7±3.4, lung phosphorylation of p38MAPK (p-p38MAPK)/p38MAPK: 0.15±0.09 vs. 0.35±0.19, lung phosphorylation of ERK1/2 (p-ERK1/2)/ERK1/2: 0.27±0.01 vs. 0.70±0.01, allP < 0.01].Conclusion HS protected against lung injury induced by severe burns in rats with a dose-dependent manner, and it was better than LR, and its possible mechanism was related with reducing the expression of p38MAPK and ERK1/2 pathway in lung tissue, increasing the level of anti-inflammatory cytokines and decreasing the release of pro-inflammatory cytokines, thus inhibiting excessive inflammation and oxidative stress injury in lung.

Objective: To explore the influence of three-level collaboration network of pediatric burns in Anhui province on treatment effects of burn children. Methods: The data of medical records of pediatric burn children transferred from Lu′an People′s Hospital and Fuyang People′s Hospital to the First Affiliated Hospital of Anhui Medical University from January 2014 to December 2015 and January 2016 to September 2017 (before and after establishing three-level collaboration network of pediatric burns treatment) were analyzed: percentage of transferred burn children to hospitalized burn children in corresponding period, gender, age, burn degree, treatment method, treatment result, occurrence and treatment result of shock, and operative and non-operative treatment time and cost. Rehabilitation result of burn children transferred back to local hospitals in 2016 and 2017. Data were processed with t test, chi-square test, Mann-Whitney U test, and Fisher′s exact test. Results: (1) Percentage of burn children transferred from January 2014 to December 2015 was 34.3% (291/848) of the total number of hospitalized burn children in the same period of time, which was close to 30.4% (210/691) of burn children transferred from January 2016 to September 2017 (χ²=2.672, P>0.05). (2) Gender, age, burn degree, and treatment method of burn children transferred from the two periods of time were close (χ²=3.382, Z=-1.917, -1.911, χ²=3.133, P>0.05). (3) Cure rates of children with mild, moderate, and severe burns transferred from January 2016 to September 2017 were significantly higher than those of burn children transferred from January 2014 to December 2015 (χ²=11.777, 6.948, 4.310, P<0.05). Cure rates of children with extremely severe burns transferred from the two periods of time were close (χ²=1.181, P>0.05). (4) Children with mild and moderate burns transferred from the two periods of time were with no shock. The incidence of shock of children with severe burns transferred from January 2014 to December 2015 was 6.0% (4/67), and 3 children among them were cured. The incidence of shock of children with severe burns transferred from January 2016 to September 2017 was 3.9% (2/51), and both children were cured. The incidences and cures of shock of children with severe burns transferred from the two periods of time were close (χ²=0.006, P>0.05). Incidence of shock of children with extremely severe burns transferred from January 2014 to December 2015 was 57.1% (32/56), significantly higher than that of burn children transferred from January 2016 to September 2017 [34.5% (10/29), χ²=3.925, P<0.05]. Shock of 25 children with extremely severe burns transferred from January 2014 to December 2015 were cured, and shock of 9 children with extremely severe burns transferred from January 2016 to September 2017 were cured. The cures of shock of children with extremely severe burns transferred from the two periods of time were close ( χ²=0.139, P>0.05). (5) Time of operative treatment of children with moderate, severe, and extremely severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, 2.065, 2.310, P<0.05). Time of operative treatment of children with mild burns transferred from the two periods of time was close (Z=-0.417, P>0.05). Costs of operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were significantly more than those of burn children transferred from January 2016 to September 2017 (Z=-3.324, t=2.167, P<0.05). Costs of operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.627, 0.808, P>0.05). (6)Time of non-operative treatment of children with mild, moderate, and severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, Z=-2.095, t=2.152, P<0.05). Time of non-operative treatment of children with extremely severe burns transferred from the two periods of time was close (t=0.450, P>0.05). Costs of non-operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were obviously higher than those of burn children transferred from January 2016 to September 2017 (Z=-2.164, t=2.040, P<0.05). Costs of non-operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.146, 1.235, P>0.05). (7) Sixty-seven burn children transferred from January 2016 to September 2017 were transferred back to local hospitals for rehabilitation under the guidance of experts of the First Affiliated Hospital of Anhui Medical University, with 25 patients in 2016 and 42 patients in 2017. Effective rehabilitation rates of burn children transferred back to local hospitals for rehabilitation in 2016 and 2017 were both 100%. Conclusions The three-level collaboration network of pediatric burns treatment in Anhui province can effectively increase cure rate of children with mild, moderate, and severe burns, reduce incidence of shock of children with extremely severe burns, shorten time of operative treatment of burn children with moderate, severe, and extremely severe burns, and time of non-operative treatment of children with mild, moderate, and severe burns, reduce treatment costs of children with moderate and severe burns, and improve rehabilitation effectiveness of children transferred from Lu′an People′s Hospital and Fuyang People′s Hospital to the the First Affiliated Hospital of Anhui Medical University.

Objective:To investigate the expression, prognostic value and potential mechanism for the role of SNHG4 in gastric cancer.Methods:The expression of SNHG4 in gastric cancer was analyzed by UALCAN database. The relationship between SNHG4 and prognosis of gastric cancer was analyzed by Kaplan-Meier Plotter. SNHG4-miRNA-mRNA regulatory network was constructed by StarBase, Targetscan, microT-CDS and Cytoscape. The target genes were analysis GO and KEGG pathway enrichment by DAVID database.Results:The expression of SNHG4 in gastric cancer was significantly higher than that in normal tissues ( P=8.882E-16) . The overall survival time of patients with high SNHG4 expression was lower than that of patients with low expression ( P=8.900E-05) . Through the construction of RNA regulatory network, we found that hsa-let-7a-5p ( P=1.02E-03) , hsa-miR-152-3p ( P=4.51E-06) , hsa-miR-204-5p ( P=6.68E-04) and hsa-miR-363-3p ( P=8.06E-03) could be used as the binding sites of SNHG4 in gastric cancer, and these four miRNAs further regulated 250 downstream target genes. Through GO and KEGG enrichment analysis of the target genes, we found that these target genes played roles in the biological process of protein phosphorylation regulation, transcription negative regulation, RNA polymerase II promoter transcription, and participated in the occurrence and development of gastric cancer by blocking or activating Wnt and other signal pathways. Conclusions:SNHG4 can be used as a potential tumor marker for gastric cancer to judge the prognosis of gastric cancer. By constructing a SNHG4-miRNA-mRNA regulatory network, the pathogenesis of gastric cancer can be studied at the molecular level. This provides a clear direction for experimental and clinical research.