Objective:To explore immunoregulatory function of long non-coding RNA Gm26688 on macrophages RAW264.7,macrophages stimulated by LPS were treated with Chinese giant salamander(CGS)collagen peptides to explore role of Gm26688 in mechanism of CGS collagen peptides in regulating inflammatory reflection.Methods:Gm26688 sequence was cloned by RT-PCR,Gm26688 localization in macrophage was analyzed by RNA-FISH;qRT-PCR was performed to detect expressions of Gm26688 and inflammatory factors after LPS and CGS bone collagen peptides treatment;expression of Gm26688 was silenced by small interfering RNA(siRNA),and qRT-PCR was used to detect effects of LPS and CGS bone collagen peptides on expression of inflammatory factors after Gm26688 interference;Western blot was used to detect protein expression of NF-κB p65 and its phosphorylation level.Results:LPS significantly promoted expression of Gm26688.RT-PCR amplified Gm26688 sequence,and RNA-FISH showed that Gm26688 was mainly expressed in nucleus.CGS bone collagen peptides inhibited expression of Gm26688 induced by LPS,and<3 kD CGS bone collagen peptides had the most significant inhibitory effect on Gm26688.CGS bone collagen peptides(<3 kD)significantly inhibited LPS-induced expressions of inflammatory factors IL-6,TNF-α and IL-1β.Interference of Gm26688 significantly down-regulated expressions of inflammatory factors IL-6 and TNF-α in LPS or LPS+CGS bone collagen peptides-stimulated RAW264.7 cells.Western blot results showed that CGS bone collagen peptides down-regulated level of NF-κB p65 phosphorylation through Gm26688.Conclu-sion:CGS bone collagen peptides inhibit LPS-induced inflammation in RAW264.7 cells via Gm26688/NF-κB pathway.

Objective:To explore immunoregulatory function of long non-coding RNA Gm26688 on macrophages RAW264.7,macrophages stimulated by LPS were treated with Chinese giant salamander(CGS)collagen peptides to explore role of Gm26688 in mechanism of CGS collagen peptides in regulating inflammatory reflection.Methods:Gm26688 sequence was cloned by RT-PCR,Gm26688 localization in macrophage was analyzed by RNA-FISH;qRT-PCR was performed to detect expressions of Gm26688 and inflammatory factors after LPS and CGS bone collagen peptides treatment;expression of Gm26688 was silenced by small interfering RNA(siRNA),and qRT-PCR was used to detect effects of LPS and CGS bone collagen peptides on expression of inflammatory factors after Gm26688 interference;Western blot was used to detect protein expression of NF-κB p65 and its phosphorylation level.Results:LPS significantly promoted expression of Gm26688.RT-PCR amplified Gm26688 sequence,and RNA-FISH showed that Gm26688 was mainly expressed in nucleus.CGS bone collagen peptides inhibited expression of Gm26688 induced by LPS,and<3 kD CGS bone collagen peptides had the most significant inhibitory effect on Gm26688.CGS bone collagen peptides(<3 kD)significantly inhibited LPS-induced expressions of inflammatory factors IL-6,TNF-α and IL-1β.Interference of Gm26688 significantly down-regulated expressions of inflammatory factors IL-6 and TNF-α in LPS or LPS+CGS bone collagen peptides-stimulated RAW264.7 cells.Western blot results showed that CGS bone collagen peptides down-regulated level of NF-κB p65 phosphorylation through Gm26688.Conclu-sion:CGS bone collagen peptides inhibit LPS-induced inflammation in RAW264.7 cells via Gm26688/NF-κB pathway.

Objective:To investigate the effects and mechanisms of glycyrrhetinic acid(GA)on the proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)of hepatocellular carcinoma SMMC-7721 cells by regulating the β-catenin/transcription factor 4(TCF4)signaling pathway.Methods:HCC cells SMMC-7721 were randomly separated into the control,L-GA,M-GA,H-GA groups(50,100,200 μmol/L GA)and the GA+LiCl group(200 μmol/L GA+40 μmol/L β-catenin activator LiCl).The effects of GA on the proliferation,migration,invasion and apoptosis of SMMC-7721 cells were detected by plate cloning,Transwell experiment and flow cytometry.WB was applied to detect the effects of GA on the expressions of β-catenin/TCF4 signaling pathway proteins(β-catenin、TCF4)and EMT(E-cadherin,vimentin)-related proteins in SMMC-7721 cells.The model of SMMC-7721 cell transplanted tumor in nude mice was established to observe the effects of GA on the growth of transplanted tumor and the expressions of β-catenin and TCF4 proteins.Results:Compared with the control group,SMMC-7721 cell clonal number,migration and invasion numbers,β-catenin,TCF4 and vmentin protein expressions in the L-GA,M-GA and H-GA groups decreased significantly(all P<0.05),while the apoptosis rate and the expression of E-cadherin protein increased(all P<0.05).Compared with the H-GA group,the numbers of clones,migration and invasion,β-catenin,TCF4 and vimentin protein expressions in the GA+LiCl group increased significantly(all P<0.05),while the apoptosis rate and E-cadherin protein expression decreased significantly(all P<0.05).Tumor-bearing nude mouse model experiment showed that the tumor weight and volume and the positive rates of β-catenin and TCF4 proteins in the GA group were significantly lower than those in the control group(all P<0.05).Conclusion:GA can significantly inhibit the proliferation,migration,invasion,and the EMT process of SMMC-7721 cells,thereby inhibiting the progression of HCC.Its mechanism may be achieved by inhibiting the β-catenin/TCF4 signaling pathway.

Objective To analyze the risk factors and pathogen drug resistance of senile stroke-associated pneumonia (SAP), and to provide references for early clinical intervention. Methods A total of 859 elderly patients with cerebral stroke admitted to our hospital from June 2018 to June 2020 were selected and divided into the study group (SAP, n=375) and the control group (no SAP, n=484) according to the occurrence of stroke associated pneumonia. Clinical data of age, gender, and other complications of the two groups were analyzed. The sputum culture and drug sensitivity test of senile SAP patients were analyzed. Results A total of 313 pathogens were detected in 375 SAP patients, including 211 strains of gram-negative bacteria (67.41%), mainly consisting of 92 Acinetobacter baumannii (29.39%), 54 Pseudomonas aeruginosa (17.25%), and 42 Klebsiella pneumoniae (13.42%), and 73 strains (23.32%) of gram-positive bacteria, mainly 62 strains of Staphylococcus aureus (19.81%). In addition, there were 29 strains of fungi (9.27%). Pseudomonas aeruginosa was highly sensitive to piperacillin/tazobactam and ceftazidime. Acinetobacter baumannii and Klebsiella pneumoniae were highly sensitive to imipenem, meropenem, and cefoperazone sodium and sulbactam sodium. Staphylococcus aureus and Enterococcus were highly sensitive to teicolanin, linezolid and vancomycin. The proportion of patients aged ≥80 years old, mechanical ventilation, bed rest and use of prophylactic antibiotics in the experimental group was significantly higher than that in the control group (P<0.05). Logistic regression analysis showed that age ≥80 years, mechanical ventilation, hypoproteinemia and use of prophylactic antibiotics were independent risk factors for SAP (P<0.05). Conclusion The main pathogens of stroke-associated pneumonia in the elderly are Pseudomonas aeruginosa, Staphylococcus aureus and Acinetobacter baumannii. It is necessary to rationally choose antibiotics according to the results of drug sensitivity. The risk factors are patients' age ≥ 80 years old, mechanical ventilation, and bed rest. Clinicians should attach great importance to the prevention of stroke-related pneumonia in the elderly.

OBJECTIVETo investigate the expression of prostaglandin transporter (PGT) in colorectal cancer (CRC) tissues and its relationship with clinicopathological features.

METHODSThe mRNA and protein levels of PGT were determined by real-time PCR, Western blot and immunohistochemical methods in cancer tissues and adjacent normal tissue from 80 patients with colorectal cancer and their relationship with clinicopathological features was analyzed.

RESULTSCompared with the adjacent normal tissue of colorectal cancer, the PGT mRNA relative expression (0.57 ± 0.33 vs. 2.33 ± 1.20) and the PGT protein expression in cancer tissues decreased significantly [PGT/GAPDH 0.45 ± 0.16 vs. 0.78 ± 0.23, integral A 718.7 ± 359.4 vs. 10412.0 ± 6423.3, average A 0.03 ± 0.01 vs. 0.12 ± 0.09, all P<0.01]. Lower mRNA and protein expressions of PGT in colorectal cancer were associated with depth of invasion T3 to T4 and TNM stage III( to IIII( (P<0.01), while not associated with gender, age, tumor location and differentiation degree (all P>0.05).

CONCLUSIONExpression levels of PGT mRNA and protein in colorectal cancer tissue are significantly down-regulation. PGT expression is associated with invasion depth and late stages.

Colorectal Neoplasms ; Down-Regulation ; Humans ; Neoplasm Invasiveness ; Neoplasm Staging ; Organic Anion Transporters ; RNA, Messenger

Objective To investigate the expression of prostaglandin transporter (PGT) in colorectal cancer (CRC) tissues and its relationship with clinicopathological features. Methods The mRNA and protein levels of PGT were determined by real-time PCR , Western blot and immunohistochemical methods in cancer tissues and adjacent normal tissue from 80 patients with colorectal cancer and their relationship with clinicopathological features was analyzed. Results Compared with the adjacent normal tissue of colorectal cancer , the PGT mRNA relative expression (0.57 ±0.33 vs. 2.33 ±1.20) and the PGT protein expression in cancer tissues decreased significantly [PGT/GAPDH 0.45 ±0.16 vs. 0.78 ±0.23, integral A 718.7 ±359.4 vs. 10412.0 ±6423.3, average A 0.03 ±0.01 vs. 0.12 ±0.09, all P<0.01]. Lower mRNA and protein expressions of PGT in colorectal cancer were associated with depth of invasion T3 to T4 and TNM stage Ⅲ to Ⅳ (P<0.01), while not associated with gender, age, tumor location and differentiation degree (all P>0.05). Conclusion Expression levels of PGT mRNA and protein in colorectal cancer tissue are significantly down-regulation. PGT expression is associated with invasion depth and late stages.

Objective To investigate the expression of prostaglandin transporter (PGT) in colorectal cancer (CRC) tissues and its relationship with clinicopathological features. Methods The mRNA and protein levels of PGT were determined by real-time PCR , Western blot and immunohistochemical methods in cancer tissues and adjacent normal tissue from 80 patients with colorectal cancer and their relationship with clinicopathological features was analyzed. Results Compared with the adjacent normal tissue of colorectal cancer , the PGT mRNA relative expression (0.57 ±0.33 vs. 2.33 ±1.20) and the PGT protein expression in cancer tissues decreased significantly [PGT/GAPDH 0.45 ±0.16 vs. 0.78 ±0.23, integral A 718.7 ±359.4 vs. 10412.0 ±6423.3, average A 0.03 ±0.01 vs. 0.12 ±0.09, all P<0.01]. Lower mRNA and protein expressions of PGT in colorectal cancer were associated with depth of invasion T3 to T4 and TNM stage Ⅲ to Ⅳ (P<0.01), while not associated with gender, age, tumor location and differentiation degree (all P>0.05). Conclusion Expression levels of PGT mRNA and protein in colorectal cancer tissue are significantly down-regulation. PGT expression is associated with invasion depth and late stages.

OBJECTIVETo explore the feasibility and clinical significance of the detection of serum neutrophil gelatinase-associated lipocalin (NGAL) in human colorectal cancer.

METHODSLevels of NGAL in serum samples from 133 healthy people, 125 colorectal polyps patients and 100 colorectal cancer patients respectively were determined by sandwich ELISA assay. Relationship of NGAL level with clinicopathological features of colorectal cancer patients was analyzed. The optimal cut-off value of serum NGAL for diagnosing colorectal cancer was determined by ROC curve and compared with CEA and CA19-9. Univariate and multivariate analyses were performed to examine the relationship of NGAL level with the prognosis of patients with colorectal cancer.

RESULTSThe median serum NGAL protein level in 100 colorectal cancer cases was 67.96 (53.30-79.86) μg/L, significantly higher than that in healthy people and colorectal polyps patients. The differences were statistically significant (all P<0.01). Serum NGAL protein level was significantly associated with tumor diameter, TNM stage, lymph node metastasis and vascular involvement (P<0.05). The optimal cut-off point of serum NGAL protein level for diagnosing colorectal cancer was 49.78 μg/L, and the sensitivity and specificity were 88% and 81% respectively. As for colorectal cancer patients with stage I, the sensitivity of serum NGAL (78.9%) was significantly higher as compared to CA19-9 (31.6%) and CEA (36.8%); as for those with stage II, the sensitivity of serum NGAL(88.0%) was also significantly higher compared to CA19-9 (48.0%) and CEA (52.0%). Kaplan-Meier analysis showed that patients with positive NGAL (≥49.78 μg/L) had worse survival than those with negative NGAL (P=0.002). Multivariate analysis showed that NGAL was an independent prognostic factor (HR=2.060, 95%CI:1.023-4.150, P=0.043).

CONCLUSIONSNGAL can be served as the novel malignant biological phenotype marker for human colorectal cancer and can be used for the risk stratification. NGAL may be an independent prognostic factor in colorectal cancer.

Acute-Phase Proteins ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Case-Control Studies ; Colorectal Neoplasms ; blood ; diagnosis ; Early Detection of Cancer ; Female ; Humans ; Lipocalin-2 ; Lipocalins ; blood ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins ; blood

Objective To explore the feasibility and clinical significance of the detection of serum neutrophil gelatinase-associated lipocalin (NGAL) in human colorectal cancer. Methods Levels of NGAL in serum samples from 133 healthy people, 125 colorectal polyps patients and 100 colorectal cancer patients respectively were determined by sandwich ELISA assay. Relationship of NGAL level with clinicopathological features of colorectal cancer patients was analyzed. The optimal cut-off value of serum NGAL for diagnosing colorectal cancer was determined by ROC curve and compared with CEA and CA19-9. Univariate and multivariate analyses were performed to examine the relationship of NGAL level with the prognosis of patients with colorectal cancer. Results The median serum NGAL protein level in 100 colorectal cancer cases was 67 . 96 ( 53 . 30-79 . 86 ) μg/L , significantly higher than that in healthy people and colorectal polyps patients. The differences were statistically significant (all P<0.01). Serum NGAL protein level was significantly associated with tumor diameter, TNM stage, lymph node metastasis and vascular involvement (P<0.05). The optimal cut-off point of serum NGAL protein level for diagnosing colorectal cancer was 49.78 μg/L, and the sensitivity and specificity were 88%and 81%respectively. As for colorectal cancer patients with stage Ⅰ, the sensitivity of serum NGAL (78.9%) was significantly higher as compared to CA19-9 (31.6%) and CEA (36.8%);as for those with stage Ⅱ, the sensitivity of serum NGAL (88.0%) was also significantly higher compared to CA19-9 (48.0%) and CEA (52.0%). Kaplan-Meier analysis showed that patients with positive NGAL (≥49.78 μg/L) had worse survival than those with negative NGAL (P=0.002). Multivariate analysis showed that NGAL was an independent prognostic factor (HR=2.060, 95%CI:1.023-4.150, P=0.043). Conclusions NGAL can be served as the novel malignant biological phenotype marker for human colorectal cancer and can be used for the risk stratification. NGAL may be an independent prognostic factor in colorectal cancer.

Objective To explore the feasibility and clinical significance of the detection of serum neutrophil gelatinase-associated lipocalin (NGAL) in human colorectal cancer. Methods Levels of NGAL in serum samples from 133 healthy people, 125 colorectal polyps patients and 100 colorectal cancer patients respectively were determined by sandwich ELISA assay. Relationship of NGAL level with clinicopathological features of colorectal cancer patients was analyzed. The optimal cut-off value of serum NGAL for diagnosing colorectal cancer was determined by ROC curve and compared with CEA and CA19-9. Univariate and multivariate analyses were performed to examine the relationship of NGAL level with the prognosis of patients with colorectal cancer. Results The median serum NGAL protein level in 100 colorectal cancer cases was 67 . 96 ( 53 . 30-79 . 86 ) μg/L , significantly higher than that in healthy people and colorectal polyps patients. The differences were statistically significant (all P<0.01). Serum NGAL protein level was significantly associated with tumor diameter, TNM stage, lymph node metastasis and vascular involvement (P<0.05). The optimal cut-off point of serum NGAL protein level for diagnosing colorectal cancer was 49.78 μg/L, and the sensitivity and specificity were 88%and 81%respectively. As for colorectal cancer patients with stage Ⅰ, the sensitivity of serum NGAL (78.9%) was significantly higher as compared to CA19-9 (31.6%) and CEA (36.8%);as for those with stage Ⅱ, the sensitivity of serum NGAL (88.0%) was also significantly higher compared to CA19-9 (48.0%) and CEA (52.0%). Kaplan-Meier analysis showed that patients with positive NGAL (≥49.78 μg/L) had worse survival than those with negative NGAL (P=0.002). Multivariate analysis showed that NGAL was an independent prognostic factor (HR=2.060, 95%CI:1.023-4.150, P=0.043). Conclusions NGAL can be served as the novel malignant biological phenotype marker for human colorectal cancer and can be used for the risk stratification. NGAL may be an independent prognostic factor in colorectal cancer.