ObjectiveTo investigate the effects of trimethylamine oxide(TMAO),a metabolite of intestinal flora,on the nuclear factor-kappa B(NF-κB)/mitogen activated protein kinase(MAPK)signaling pathway in human glomerular mesangial cells HMC and renal tubular epithelial cells HK-2.MethodsHMC and HK-2 cells were treated with TMAO and the control group was treated with TMAO for 0 h. The mRNA transcription levels of MCP-1,IL-6,TNF-α and IL-1β were detected by Real-time PCR,while the expression of NF-κB and MAPK signaling pathway-related proteins by Western blot. HMC and HK-2 cells were treated with NF-κB inhibitor BAY11-7082 to verify the pro-inflammatory effects of TMAO.ResultsCompared with the control group,the expression levels of inflammatory factors MCP-1,IL-6,TNF-α and IL-1βmRNA significantly increased in the TMAO-treated group(t = 33. 349,P = 0. 001),indicating that TMAO had a significant pro-inflammatory effect on glomerular mesangial cells and renal tubular epithelial cells. TMAO activated NF-κB and MAPK signaling pathway in HMC and HK-2 cells,causing inflammatory response and damage to the kidney.ConclusionThe increase of TMAO level can activate the NF-κB/MAPK signaling pathway in HMC and HK-2 cells,stimulate the release of inflammatory cytokines from the both kinds of cells,which leads to the occurrence and development of renal injury.

Objective To investigate the clinical application value of endotoxin and (1-3)-β-D glucan detection in the patients with acute pancreatitis(AP).Methods The MB-80 microbiology rapid dynamic detection system was applied to detect the concentration of plasma LPS and (1-3)-β-D glucan in the patients with AP.Then the results were analyzed.Results Among 184 cases of AP complicating peripancreatic abscess and bilateral lung infection,endotoxin G-lipopolysaccharides positive was in 23 cases,the average value was 394.07 pg/mL,the positive rate was 10.87%,and 15 cases of Gram-negative bacterial infection were verified by the microorganism culture.Among 175 cases of AP complicating peripancreatic abscess and bilateral lung infection,67 cases were(1-3)-β-D glucan positive,the average value was 93.22 pg/mL,the positive rate was 38.29%,which were with the positive rate of serum(1-3)-β-D glucan in the non-AP group,the difference was statistically significant(P<0.05),the microorganism culture or smear verified that 22 cases were fungal infection.Conclusion Plasma endotoxin and (1-3)-β-D glucan detection provides the laboratory biological indicator and is conducive to the assessment of AP infection severity and clinical medication.

Objective To study the application value of G-lipopolysaccharides(G-LPS) detection in patients with traumatic brain injury(TBI).Methods From Jan.2013 to Jun.2014,plasma G-LPS of TBI-patients and non-patients in this hospital were detected by using MB-80 microbiology analyzer and compared.Results of pathogenic microbiological detection of G-LPS positive TBI patients were analyzed,and G-LPS levels,detected before and after treatment,were also analyzed.Results G-LPS positive rate of TBI patients(33.33%) was higher than the 13.34% of non-TBI patients(P<0.05).After treatment,G-LPS level in G-LPS positive TBI patients significantly decreased(P<0.05).The common pathogenic bacteria,causing infection in TBI patients,included Acinetobacter bauman,Klebsiella pneumoniae and Escherichia coli,and the most common infection diseases included respiratory tract infections and urinary tract infections.Conclusion G-LPS detection could be used for the early detection of secondary infection in TIB patients,with significance for guiding clinical treatment.

Objective To study the detection rate of contrast-enhanced ultrasound (CEUS) with different perfusion methods on rabbit VX2 small hepatocellular carcinoma less than 10.0 mm. Methods VX2 tumor cells were inoculated into the subcutaneous tissue of New Zealand rabbit′s thigh. Then the plant tumor were cut into small blocks under sterile conditions and transplanted into hepatic parenchyma in 30 New Zealand rabbits. The contrast media was injected through peripheral vein by single and double perfusion methods. The detection rate of two methods were compared. Results There were 41 hepatocellular carcinoma lesions in the 30 rabbits. There were 15 lesions with size between 3.0 mm and 5.0 mm, and 26 lesions between 5.0 mm and 10.0 mm in diameter. On CEUS, the VX2 tumor presented fast-in and fast-out pattern. In arterial phase, the lesion was enhanced rapidly. In portal venous phase, contrast began to wash out from the carcinoma. In delay phase, the enhancement of lesion was signiifcantly lower than the surrounding normal liver parenchyma. A total of 32 lesions were detected by single perfusion method, including 7 lesions ranging 3.0-5.0 mm and 25 lesions ranging 5.0-10.0 mm. A total of 39 lesions were detected by double perfusion method, including 13 lesions ranging 3.0-5.0 mm and 26 lesions ranging 5.0-10.0 mm. The detection rate of micro-hepatocellular carcinoma by single and double perfusion method was 78% and 95% respectively. The difference was statistically signiifcant (χ2=5.150, P=0.023). The detection rate of 3.0-5.0 mm lesions by single and double perfusion method was 47%and 87%, respectively. The difference was statistically signiifcant ( χ2=5.400, P=0.025). The detection rate of 5.0-10.0 mm lesions by single and double perfusion method was 96% and 100%, respectively. There was no statistically signiifcant difference (χ2=1.020, P=0.500). Conclusion The double perfusion method greatly promotes the detection of micro hepatocellular carcinoma, especially for the lesions less than 5.0 mm in diameter.

Objective To observe the effects of live combined bifidobacterium,lactobacillus and enterococcus powder on immunoglobulin E (IgE) and interleukin-17 (IL-17) in atopic children with bronchiolitis.Methods Sixty cases of atopic children with bronchiolitis were randomly divided into the therapy group (30 cases) and the control group (30 cases).Twenty-five healthy children were enrolled as the healthy control group.Both the therapy group and the control group were given traditional therapy.The therapy group received live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months.The change of IgE and IL-17 levels were observed during the acutestage,remission stage and after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2months.Results (1) The levels of IgE and IL-17 of therapy group[(132.36 ±9.50) μg/L and (77.76 ±7.95)μg/L] during acute stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ±4.79) μg/L] (all P ＜0.001).The levels of IgE and IL-17 of control group [(128.83 ± 8.06) μg/L and (76.61 ±6.18) μg/L] during remission stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ± 4.79) μg/L] (all P ＜ 0.001).(2) The levels of IgE of therapy group (56.67 ± 9.20)μg/L after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than those in the control group (70.50 ± 11.38) μg/L (P ＜ 0.001).The levels of IL-17 of therapy group [(49.63 ± 6.35) μg/L] at the time after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than these in the control group (54.77 ± 6.33) μg/L (P =0.003).Conclusion Receiving live combined bifidobacterium,lactobacillus and enterococcus powder for two months can decrease the IgE and IL-17 levels in atopic children with bronchiolitis.

Objective To investigate the clinical significance and changes of serum interleukin-4 (IL-4),interferon-γ(IFN-γ) and immunoglobulin E (IgE) levels in infants with spasmodic laryngitis.Methods Serum samples were obtained from 50 children with spasmodic laryngitis and 30 healthy children.The changes of IL-4,IFN-γand IgE levels in serum when hospital admission (acute stage),before discharge (the stage of clinical symptom disappeared) and after 2 months of follow-up were observed.Enzyme-linked immunosorbent assay was used to determine the levels of IL-4 and IFN-γ in serum.Serum IgE level was determined with enzyme-linked fluoroimmuneassay.Results The serum level of IL-4 in patients with spasmodic laryngitis during acute stage were much higher than that in normal control group[(20.65 ±5.87) ng/L vs(8.23 ±2.71) ng/L,t =5.536,P ＜0.05].It was decreased during the stage of clinical symptom disappeared and significantly lower than that during acute stage[(11.07 ± 2.93) ng/L vs (20.65 ± 5.87) ng/L,F =5.258,P ＜ 0.05].The serum level of IFN-γ in patients with spasmodic laryngitis during acute stage were much lower than that in normal control group[(61.38 ±6.36) ng/L vs (108.42 ± 13.56) ng/L,t =4.831,P ＜ 0.05],and it was increased during the stage of clinical symptom disappeared [(95.12 ± 11.63) ng/L] and significantly higher than that during acute stage(F =4.239,P ＜ 0.05).There was no difference between the patients at clinical symptom disappeared stage and normal control group (P ＞ 0.05).Two months after discharge,there was no difference in serum INF-γlevel between the patients [(75.68 ±6.29) ng/L] and that in control group (P ＞ 0.05).The serum level of IgE in patients with spasmodic laryngitis during acute stage were much higher than that in normal control group [(136.63 ± 17.86) ng/L vs (47.16 ± 6.83) ng/L,t =6.685,P ＜ 0.05].It was decreased during the stage of clinical symptom disappeared [(88.07 ± 11.83) μg/L] and significantly lower than that during acute stage(F =5.182,P ＜ 0.05),but higher than that of normal control group (P ＜ 0.05).Two months after discharge [(86.72 ± 7.25) μg/L] it was still higher than that in control group (P ＜ 0.05).Conclusions There is TH1/TH2 imbalance in infants with spasmodic laryngitis.Maybe spasmodic laryngitis is one of the clinical manifestations of atopic diseases.IL-4,IFN-γand IgE may play important roles in immunologic pathogenesis.

Objective:To study the value of metastatic lymph node radio (rN) and pathological lymph node stage (pN) in evaluating the prognosis of patients after radical gastric cancer.Methods:The clinicopathological data of 491 patients who underwent radical gastrectomy in Tantai Yantaishan Hospital from Jan. 2013 to Dec. 2017 were retrospectively analyzed. X-tile software was used to group the metastatic lymph node radio by rN. According to the number of lymph node metastasis, pN stage was performed. The correlation between metastatic lymph node radio and other clinicopathological factors was assessed. The metastatic lymph node radio and the pathological lymph node stage in evaluating the prognosis of patients after radical gastric cancer were compared.Results:(1) X-tile analysis showed that the best cut-off values for the metastatic lymph node radio in this study were 0.14 (14%) and 0.63 (63%) . (2) According to the cut-off value, the 491 patients included in the study were divided into rN1 (256 cases) , rN2 (160 cases) , and rN3 (75 cases) three subgroups. The results of the analysis of differences showed that there were significant differencesbetween the groups in terms of tumor diameter, tumor location, surgical resection range, stage, lauren classification, degree of differentiation, pT, pN, vascular cancer emboulus, nerve invasion, and pathological TNM staging groups. (3) Comparison of rN and pN staging in evaluation of the prognosis of patients after radical gastric cancer: ①Kaplan-Meier survival analysis results showed that rN was better than pN. ② Both single factor and multivariate cox analysis showed that rN was an independent risk factor for the prognosis of gastric cancer. In univariate analysis, rN group HR=3.18 (95% CI 2.63-3.84, P<0.001) , pN stage HR=1.88 (95% CI 1.66-2.15, P<0.001) ; rN group HR=2.21 in multivariate analysis (95% CI 1.73-2.82, P<0.001) , pN staging HR=1.31 (95% CI 0.95-1.79, P=0.095) . ③The time-dependent ROC analysis showed that the prognostic ability of rN was better than pN staging before 52 months of postoperative follow-up, and pN staging was more advantageous after 52 months. ④The Lauren classification was used as a stratification factor for stratified analysis. The Kaplan-Meier survival curve indicated that rN was better than pN staging in intestinal, mixed and diffuse gastric cancer, and the AUC curve showed the prediction of rN in patients with mixed and diffuse gastric cancer was better than pN staging, while pN staging performance was slightly better in patients with intestinal gastric cancer. Conclusions:rN is an independent factor affecting the prognosis of patients after radical gastric cancer surgery. When judging the prognosis of patients within 52 months after radical gastric cancer, rN has a better prognostic value than pN. In patients with mixed and diffuse gastric cancer in the Lauren classification, rN shows better prognostic value.