Objective To investigate the release of enterogenic and hepatogenic high mobility group protein B1(HMGB1)through exosomes and its regulatory pathway.Methods We used wild-type(WT)and ASC-/-mice for this study.We randomly selected five mice per group from each strain and fed them either a normal diet(ND)or a high-fat diet(HFD)for eight weeks.The control group consisted of WT mice fed with the normal diet;the HFD group were WT mice with the HFD;the microflora disturbance(MD)group were ASC-/-mice fed with the normal diet;the high-lipid microflora disturbance(HLMD)group were ASC-/-mice with HFD.We used confocal microscopy to detect the co-localization of liver and intestinal exosome markers with HMGB1.We then measured the expression level of HMGB1 content in exosomes by Western blotting and PCR.The AML12 cells were treated with palmitic acid(PA)and lipopolysaccharide(LPS)for 24 h to build an in vitro model.We also detected HMGB1/CD63 levels using Western blotting.To understand the regulatory mechanism of exosome release,we employed siRNA intervention.Results The secretion of exosomes increased significantly in HFD group compared with control group[(3.5±0.2)ng/ml vs.(1.1±0.3)ng/ml,P<0.05],HLMD group compared with those in MD group[(3.2±0.2)ng/ml vs.(1.9±0.4)ng/ml,P<0.05].Using immunofluorescence detection,we observed increased co-localization of exosome markers(ALP or VPS16)with HMGB1 in HFD group compared with control group.We also observed this in AML12 cells treated with PA and LPS compared with blank control.The PCR data showed that HMGB1 in hepatocyte exosomes was higher in HFD group compared with control group(41.5±10.2 vs.1.3±0.3,P<0.05),HLMD group was significantly higher than that in MD group(48.6±7.2 vs.1.5±0.5,P<0.05).TLR4 expression was higher in HFD group compared with control group(13.8±6.2 vs.2.8±0.9,P<0.05),HLMD group compared with MD group(22.6±4.1 vs.2.5±1.5,P<0.05).In intestinal mucosal cells,the co-location of HMGB1 and exosome marker CD63 was significantly higher in HFD group compared with control group(0.6±0.2 vs.0.4±0.1,P<0.05),and HLMD group compared with MD group(0.9±0.2 vs.0.5±0.1,P<0.05).In vitro,the HMGB1 of exosomes was increased in endotoxin group(5.1±0.8)and high lipid endotoxin group(5.5±0.7)compared with control group(3.8±0.6,P<0.05).On the other hand,the HMGB1 of exosomes in the cell siRNA intervention group was not increased compared with control group(3.7±0.6 vs.3.8±0.6,P>0.05).Conclusion HMGB1 is released by exosomes in hepatocytes and intestinal cells,and regulated by Toll-like receptor 4(TLR4)under a high-fat diet and intestinal flora disorder,which may be one of the contributing factors in promoting the development of steatohepatitis.

Objective: To study the expression and effect of small nuclear ribonucleoprotein-associated protein B (SNRPB) on proliferation and metastasis of liver cancer tissues and cells. Methods: The bioinformatics database starBase v3.0 and GEPIA were used to analyze the expression of SNRPB in liver cancer tissue and normal liver tissue, as well as the survival and prognosis of liver cancer patients. The expression of SNRPB mRNA and protein in liver cancer cell lines were analyzed by qRT-PCR and Western blot. RNA interference technique (siRNA) was used to determine SNRPB protein expression down-regulation. The proliferation effect on hepatocellular carcinoma cells was observed by MTT assay. Transwell invasion and migration assay was used to detect the changes in the metastatic ability of liver cancer cells after SNRPB down-regulation. Western blot was used to detect the changes of epithelial mesenchymal transition (EMT) markers in liver cancer cells after down-regulation of SNRPB expression. Data were compared between two groups and multiple groups using t-test and analysis of variance. Results: The expression of SNRPB was significantly higher in liver cancer tissue than normal liver tissue, and its expression level was correlated with the prognosis of liver cancer patients. Compared with the immortalized hepatocyte LO(2), the expression of SNRPB was significantly increased in the liver cancer cells (P < 0.01). siRNA-SNRPB had significantly inhibited the expression of SNRPB mRNA and protein in liver cancer cells. MTT results showed that the absorbance value was lower in SNRPB knockdown group than negative control group, and the difference at 96 h after transfection was most significant (P < 0.01). Transwell assay results showed that compared with the negative control group, the SNRPB knockdown group (MHCC-97H: 121.27 ± 8.12 vs. 46.38 ± 7.54; Huh7: 126.50 ± 6.98 vs. 41.10 ± 8.01) invasion and migration (MHCC-97H: 125.20 ± 4.77 vs. 43.18 ± 7.32; Huh7: 132.22 ± 8.21 vs. 38.00 ± 6.78) ability was significantly reduced (P < 0.01) in liver cancer cells. Western blot showed that the expression level of epithelial phenotype marker E-cadherin was decreased after down-regulation of SNRPB, while the expression levels of mesenchymal phenotype markers N-cadherin and vimentin was increased, suggesting that down-regulation of SNRPB inhibited EMT in liver cancer cells. Conclusion: SNRPB expression is significantly increased in liver cancer tissues and cells, and it is involved in regulating the proliferation, metastasis and EMT of liver cancer cells.
Carcinoma, Hepatocellular/genetics* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms/genetics* ; snRNP Core Proteins

Thrombospondin 4 (THBS4), a member of the THBS family, is a protein secreted by the extracellular matrix and is involved in regulating various physiological processes, such as cell proliferation, adhesion and angiogenesis. Recent studies have shown that the inflammation stimulates THBS4 production and induces the adhesion and accumulation of macrophages. Our previous study confirmed that THBS4 acts as an oncogene in hepatocellular carcinoma (HCC), the effect of THBS4 on the immune microenvironment of HCC remains unclear. This study aims to analyze the role of THBS4 in promoting the metastasis of HCC cells by inducing M2-type polarization of tumor-associated macrophages. We simulate the tumor microenvironment through HCC conditioned medium (HCM) and found that the expression of THBS4 in macrophages increased in a time-dependent manner under the action of HCM (P<0.05); THBS4 knockdown promotes the expression of M1 macrophages markers IL-1β and CD86 (P<0.01), while the expression of M2-type markers IL-10 and CD206 were decreased (P<0.01). Transwell co-culture assay was used to further detect the effect of THBS4-induced M2-type macrophages on HCC metastasis. Results from co-culture of THBS4-downregulated M2 macrophages with HepG2 cells showed that THBS4-downregulated M2-TAMs significantly inhibited the invasion and migration ability of HepG2 cells (all P < 0.01). In conclusion, the tumor microenvironment promotes the expression of THBS4 in macrophages, and THBS4 may promote the invasion and metastasis of HCC cells by inducing M2-type polarization of macrophages. This study provides some new experimental basis for exploring the establishment of THBS4-induced HCC immune microenvironment.

PURPOSE: Hemodynamically unstable patients with pelvic fractures still represent a challenge to trauma surgeons and have a very high mortality. This study was designed to explore the effect of the interventions of direct preperitoneal pelvic packing for the hemodynamically unstable pelvic fractures. METHODS: This retrospective study enrolled 67 cases of severe pelvic fractures with unstable hemodynamics from October 2011 to December 2019. All patients presented in our emergency center and received preperitoneal pelvic packing were included in this study. The indication was persistent systolic blood pressure ≤90 mmHg during initial resuscitation and after transfusion of two units of red blood cells. Patients with hemodynamic stability who need no preperitoneal pelvic packing to control bleeding were excluded. Their demographic characteristics, clinical features, laboratory results, therapeutic interventions, adverse events, and prognostic outcomes were collected from digital information system of electronic medical records. Statistics were described as mean ± standard deviation or medium and analyzed using pair sample t-test or Mann-Whitney U-test. RESULTS: The patients' average age was 41.6 years, ranging from 10 to 88 years. Among them, 45 cases were male (67.2%) and 22 cases were female (32.8%). Significant difference was found regarding the systolic blood pressure (mmHg) in the emergency department (78.4 ± 13.9) and after preperitoneal pelvic packing in the surgery intensive care unit (100.1 ± 17.6) (p < 0.05). Simultaneously, the arterial base deficit (mmol/L) were significantly lower in the surgery intensive care unit (median -6, interquartile range -8 to -2) than in the emergency department (median -10, interquartile range -14 to -8) (p < 0.05). After preperitoneal pelvic packing, 15 patients (22.4%) underwent pelvic angiography for persistent hypotension or suspected ongoing haemorrhage. The overall mortality rate was 29.5% (20 of 67). CONCLUSIONS Preperitoneal pelvic packing, as a useful surgical technique, is less invasive and can be very efficient in early intra-pelvic bleed control.

ObjectiveThere are few studies on the correlation between the concentration of oncoembryonic antigen associated cell adhesion molecule 1(CEACAM1) and osteonecrosis of the femoral head (ONFH). The purpose of this study was to investigate the value of CEACAM1 in the early diagnosis of ONFH and the monitoring of the disease by detecting the CEACAM1 concentration in the serum of patients with ONFH and healthy subjects respectively.Methods95 patients, who were hospitalized and diagnosed as ONFH in the Department of No.3 Orthopaedic Ward, the First Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2016 to November 2016, were selected as the experimental group. In addition, 56 genders and age-matched healthy subjects in our hospital were selected as the control group. The peripheral venous blood was taken and separated by a centrifuge. Their CEACAM1 concentrations were measured by enzyme linked immunosorbent assay (ELISA). The differences in CEACAM1 concentrations were analyzed between the two groups, and between patients with ONFH before (ARCO stage I or II) and after (ARCO stage III or IV) collapse as well.Results①The concentration of CEACAM1 in the experimental group was significantly lower than that in the control group [(6.11±2.07)ng/mL vs (7.21±3.76)ng/mL, P=0.022]. ②The concentration of CEACAM1 in Arco stage II[(7.33±1.90) ng/mL] was significantly higher than that in stage III [(6.08±2.26) ng/mL], P=0.037.③The difference of CEACAM1 concentration between before(stage II) and after collapse (stage III or stage IV) was statistically different [(7.33±1.90)ng/mL vs (5.86±2.02)ng/mL, P=0.007].④ROC curve analysis showed that the area under the curve was 0.710 (0.608-0.798), the sensitivity was 71.79%, the specificity was 58.82%, and the cut off value was ≤ 6.757ng/mL in the diagnosis of collapse of ONFH.ConclusionThe concentration of serum CEACAM1 can be used as a biochemical marker to assist in the diagnosis and monitoring of ONFH, which can provide reference for early diagnosis and monitoring of ONFH.

Newcastle disease virus (NDV) and Salmonella Pullorum have significant damaging effects on the poultry industry, but no previous vaccine can protect poultry effectively. In this study, a recombinant-attenuated S. Pullorum strain secreting the NDV hemagglutinin-neuraminidase (HN) protein, C79-13ΔcrpΔasd (pYA-HN), was constructed by using the suicide plasmid pREasd-mediated bacteria homologous recombination method to form a new bivalent vaccine candidate against Newcastle disease (ND) and S. Pullorum disease (PD). The effect of this vaccine candidate was compared with those of the NDV LaSota and C79-13ΔcrpΔasd (pYA) strains. The serum hemagglutination inhibition antibody titers, serum immunoglobulin G (IgG) antibodies, secretory IgA, and stimulation index in lymphocyte proliferation were increased significantly more (p < 0.01) in chickens inoculated with C79-13ΔcrpΔasd (pYA-HN) than with C79-13ΔcrpΔasd (pYA) but were not significantly increased compared with the chickens immunized with the LaSota live vaccine (p > 0.05). Moreover, the novel strain provides 60% and 80% protective efficacy against the NDV virulent strain F48E9 and the S. Pullorum virulent strain C79-13. In summary, in this study, a recombinant-attenuated S. Pullorum strain secreting NDV HN protein was constructed. The generation of the S. Pullorum C79-13ΔcrpΔasd (pYA-HN) strain provides a foundation for the development of an effective living-vector double vaccine against ND and PD.
Animals ; Antibodies ; Bacteria ; Chickens ; Hemagglutination ; HN Protein ; Homologous Recombination ; Immunoglobulin A, Secretory ; Immunoglobulin G ; Lymphocytes ; Methods ; Newcastle disease virus ; Newcastle Disease ; Plasmids ; Poultry ; Salmonella ; Suicide ; Vaccines

Objective To investigate the effects of different bolus and swallow patterns on esophageal manometry in patients with gastroesophageal reflux disease by high resolution manometry .Methods Patients with gastroesophageal reflux disease questionnaire score of more than 8 points and positive 24-hour pH monitoring were included in the study .All the patients were detected by liquid swallow ,solid swallow and continuous swallow .The parameters and comprehensive diagnosis were in accordance with the Chicago Standard .Results A total of 42 patients with gastroesophageal reflux disease were enrolled . Compared with the dynamic parameters of liquid swallow ,the residual pressure of upper esophageal sphincter [(11 .07 ± 3 .97 ,5 .29 ± 3 .36)mmHg] decreased ,the distal latency [(6 .28 ± 1 .87 ,8 .98 ± 2 .25)s] ,and lower esophageal sphincter relaxation time [(7 .79 ± 0 .98 ,10 .69 ± 13 .04)s] prolonged significantly (all P<0 .05) .In the comprehensive diagnosis of esophageal motility ,compared with liquid swallow (38 .1% ) , continuous liquid swallow showed a more sensitive positive diagnostic rate of ineffective esophageal motility (IEM) (63 .2% ) ,with a significant difference (P=0 .008) .Compared with that of liquid swallow ,the diagnostic rate (45 .2% ) of IEM by the solid swallow did not differ significantly (P=0 .581) . Among the ineffective contraction ,the rate of failed contraction (44.3% ) of solid swallow was higher than that of liquid swallow (22 .6% ) .Conclusion Solid swallow is more likely to induce severe esophageal hypomotility disorders than liquid swallow.Continuous swallow has more sensitivity in the diagnosis of IEM.Therefore.it can be used as a supplement to routine manometry in patients with eastroesophageal reflux disease.

Objective To investigate the induction of microRNA (microRNA-106a,miR-106a)on the peritoneal metastasis of human gastric cancer cell BGC-823 by regulating matrix metalloproteinase inhibitor 2 (tissue inhibitor of metalloproteinases 2,TIMP2).Methods Human gastric cancer cell line BGC-823 was cultured to the logarithmic growth phase. The cells were divided into three groups: BGC-823, BGC-823/anti-miR-106a (antagomir)and BGC-823/negative control.Real-time PCR was used to identify the effect of antagomir.Transwell assay was used to detect the cell migratory and invasive abilities of these three groups in vitro .With small incision, the cells were injected into the abdominal cavity of nude mice to prepare a xenograft model.The animals were divided into two groups:miR-antagomir and miR-NC.The tumor growth in the nude mice was generally observed and estimated.Immunohistochemistry and Western blot methods were used to detect the expression of metastasis-associated protein TIMP2 on the several abdominal organs.Results The expression level of miR-106a was down- regulated in BGC-823/anti-miR-106a group,with the fold change of 0.05±0.01,which was significantly different from that in NC group (t=-18.001,P＜0.001).In vitro exogenously silencing of miR-106a gene,the numbers of invasive and migratory cells in BGC-823/anti-miR-106a group were both significantly lower than those in BGC-823 and BGC-823/negative control groups (P＜0.001).In vivo xenograft model showed that the down-regulation of miR-106a weakened the peritoneal metastasis ability of BGC-823 cells in nude mice abdominal cavity,which was reflected by the decrease of tumor number and tumor size.With the inhibition of miR-106a,the expression of TIMP2 in miR-antagomir group was significantly higher than that in miR-NC group (P＜0.05).Conclusion BGC-823 cell has the tumorigenicity in nude mice.Silencing of miR-106a inhibits gastric cancer cell metastasis, which suggests that it has the oncogenic function.MiR-106a may induce the strengthened peritoneal metastasis ability of BGC-823 cell through acting on TIMP2.

Objective To survey the genetic polymorphism distribution of cytochrome P(CYP) 3A4 * 1 B,CYP 3A4 * 1G,CYP 3A5 * 3,human leukocyte antigen(HLA)-B * 1502 gene associated with carbamazepine therapy in healthy volunteers of Han Chinese.Methods A total of 187 Han Chinese healthy volunteers were randomly selected and their related genotypes were analyzed by fluorescence in situ hybridization.Statistical analysis was performed.Results CYP 3A4 * 1 B loci were all AA genotype (100％).The genotypes of CC,CT and TT in CYP 3A4 * 1G gene locus accounted for 62.57％,34.22％ and 3.21％,respectively;GG,AG and AA genotypes in CYP3A5 * 3 locus accounted for 58.29％,34.76％ and 6.95％;the CC and CG genotypes in HLA-B * 1502 locus accounted for 89.30％ and 10.70％ respectively,but no GG genotype was found.Conclusion The CYP 3A4 * 1 B,CYP3A4 * 1 G,CYP 3A5 * 3 and HLA-B * 1502 gene polymorphisms were more common in Chinese Han healthy volunteers,the mutation rate was high,and some of the results were consistent with the previous researches,also there was a significant difference in the results with some other populations.

The purpose of this study was to investigate the etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population. The clinical data of 142 patients with chronic pancreatitis were retrospectively studied. All patients were of Chinese nationality and hospitalized from January 2008 to December 2011. Their ages ranged from 14 to 76 years, with a mean of 43 years. Of 142 patients, there were 72 cases of obstructive chronic pancreatitis (50.70%), 19 cases of alcoholic chronic pancreatitis (13.38%), 14 cases of autoimmune pancreatitis (9.86%) and 37 cases of undetermined etiology (26.06%). Pathologically, the average inflammatory mass diameter was 3.8 ± 3.3 cm, biliary obstruction occurred in 36 cases, gall stones in 70 cases, calcification in 88 cases, ductal dilatation in 61 cases, side branch dilatation in 32 cases, ductal irregularity in 10 cases, lymphocytic inflammation in 23 cases, obliterative phlebitis in 14 cases, extra pancreatic lesion in 19 cases and fibrosis in 142 cases. Location of pancreatic lesion in the region of head (n=97), neck (n=16), body (n=12), tail (n=15) and whole pancreas (n=2) influenced the choice of surgical procedures. Ninety-four patients (66.20%) received surgical treatment and 33.80% received other treatments. After operation, 80.85% of 94 patients experienced decreased pain, and 8.51% of 94 showed recovery of endocrine function but with a complication rate of 12.77%. All the operations were performed successfully. According to the pain scale of European Organization for Research and Treatment of Cancer (QLQ-C30) a decrease from 76 ± 22 to 14 ± 18 was observed. Etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population vary from others.
Adolescent ; Adult ; Aged ; Autoimmune Diseases ; epidemiology ; therapy ; China ; epidemiology ; Cholestasis ; epidemiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis, Alcoholic ; epidemiology ; therapy ; Pancreatitis, Chronic ; etiology ; pathology ; therapy ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Young Adult