The purpose of this study was to explore the protective effect of glycyrrhiza polysaccha-ride on oxidative stress-induced liver cell damage in broiler chickens,and to study its mechanism of action.Primary broiler liver cells cultured in vitro were divided into blank control group,H2 O2 in-duction group,glycyrrhiza polysaccharide group and H2O2+glycyrrhiza polysaccharide group;CCK-8 reagent was used to detect liver cell activity in broiler chickens,JC-1 and DAPI staining kits were used to detect cell apoptosis,relevant kits were used to detect GSH-Px,SOD,CAT,MDA content,and T-AOC levels.qRT-PCR was used to detect antioxidant genes Nrf2,Keap-1,GPx1,SOD,CAT and inflammatory genes TLR4,MyD88,NF-κB,TNF-α、IL-6,IL-1β、IL-4 and IL-10.The results showed that compared with the H2O2 induced group,glycyrrhiza polysaccharide pretreat-ment could significantly enhance liver cell activity and reduce cell apoptosis in broiler chickens;compared with the H2O2 induced group,glycyrrhiza polysaccharide pretreatment could significant-ly increase cell GSH-Px,SOD,CAT,MDA content and T-AOC levels,and inhibit MDA genera-tion;qRT-PCR detection showed that glycyrrhiza polysaccharide could significantly increase the relative expression levels of Nrf2,Keap-1,GPx1,SOD,CAT,IL-4 and IL-10,while significantly re-ducing MyD88,NF-κB,TNF-α,IL-6 and IL-1β.The results indicated that glycyrrhiza polysaccha-ride enhance the activity of antioxidant enzymes by activating the Nrf2 signaling pathway and in-hibit TLR4 signaling pathway transduction,significantly improving oxidative stress damage in broiler liver cells.

Objective:To explore the clinicopathological and genetic characteristics of sporadic medullary thyroid carcinoma (MTC) and new therapeutic targets for sporadic MTC.Methods:Based on family and personal disease history, we identified 32 sporadic MTC who underwent surgical resection from Jan 2010 to Dec 2022. Clinicopathological and immunohistochemical features were analyzed in all patients, while 6 of them were subject to the whole exome sequencing (WES).Results:Compared with those of low-grade sporadic MTC, patients with high-grade tumors were more likely to have lymph node metastasis at presentation ( χ2=4.428, P=0.040); less likely to be cured by biochemical treatment ( χ2=4.072, P=0.044). Pathological grading scheme, biochemical cure, and TNM stage were independent risk factors of disease free survival. WES was performed on 6 pairs of normal tissues. We screened RET and RAS as driver mutations, and the mutation ratio was 3/6 respectively. Patients with RET or RAS mutations had no recurrence. In addition, we detected PDGFRA somatic mutation, with a mutation ratio of 1/6. Conclusions:For sporadic MTC cases, the pathological grading system has important prognostic value, and RET and RAS somatic mutations are the main driver mutations. PDGFRA are potential therapeutic targets for sporadic MTC.

Objective:To investigate the prevalence and clinical characteristics of Mycoplasma pneumoniae( Mp) genotypes and subtypes in children in Tianjin. Methods:Children with pneumonia admitted to Tianjin Children′s Hospital from December 2017 to December 2019 were selected as the research objects. Bronchoalveolar lavage fluid was collected by fiberoptic bronchoscopy. The positive samples were detected by real-time fluorescent quantitative PCR and Mp culture. PCR-restriction fragment length polymorphism(RFLP) and multiple variable number tandem repeats were used for genotyping. Detailed clinical and laboratory data were collected for all cases. Results:The results of RFLP showed that there were 138 cases (78.9%) of typeⅠand 37 cases (21.1%) of type Ⅱ; 37 cases of type M3-5-6-2, including six subtypes B, G, M, S, V and Y; 138 cases of M4-5-7-2 were detected, including seven subtypes of E, J, P, U, X, Z and a. In M3-5-6-2 type, there were 1 case of P1-Ⅰtype (2.7%), 36 cases of P1-Ⅱtype (97.3%), 137 cases of P1-Ⅰ type (99.2%) and 1 case of P1-Ⅱ type (0.7%) in M4-5-7-2 type. There was no significant difference in genotype distribution among different age groups. There were statistical differences in the distribution of four seasons among the 13 genotypes of B, G, M, S, V, Y and E, J, P, U, X, Z, a. All Mp infected children had symptoms of fever and cough. The hospitalization time, fever duration, high fever (>39℃), cough duration, skin changes, digestive system symptoms and liver function injury rate of P1-Ⅰ/M4-5-7-2 pneumonia children were higher than those of P1-Ⅱ/M3-5-6-2 pneumonia children, but the difference was not statistically significant. The WBC count of P1-Ⅱ/M3-5-6-2 types was higher than that of typeⅠand M4-5-7-2; the LDH of P1-Ⅰ/M4-5-7-2 was higher than that of Ⅱ and M3-5-6-2, with statistical difference. There was no significant difference in the incidence of inflammatory consolidation, atelectasis, pleural thickening and pleural effusion among different genotypes. Conclusions:Mp infection in children with pneumonia in Tianjin is mainly P1-Ⅰ/ M4-5-7-2, and P1-Ⅱ is on the rise. P1-Ⅰ and M4-5-7-2 were associated with fever and severe symptoms.

Objective:To analyze the pathogenic bacteria and epidemiological characteristics in children with respiratory tract infection in Tianjin area.Methods:Retrospective case analysis was performed on 2 392 hospitalized children in the wards of respiratory diseases, intensive care unit and special care ward of Tianjin Children′s Hospital from June 2018 to May 2019. Thirteen pathogenic bacteria in deep sputum and bronchoalveolar lavage fluid samples were detected by loop-mediated isothermal amplification. The laboratory data and clinical characteristics of the infected children were analyzed, and the comparison between groups was performed by t test or χ 2 test. Results:Among 2 392 cases, 1 407 were males and 985 females. There was no significant difference in the detection rate between males and females (72.5% (1 020/1 407) vs.74.2% (731/985), χ 2=0.87, P=0.35). A total of 1 751 strains and 12 kinds of positive respiratory pathogens were detected, with a detection rate of 73.2%. Among them, 913 (38.2%) strains were Mycoplasma pneumoniae (MP), 514 (21.5%) were Streptococcus pneumoniae (Sp), 381 (15.9%) were Methicillin-resistant Staphylococcus aureus (MRSA) and 279 (11.7%) were Hemophilus influenzae (Hi). There was significant difference in the detection rate of pathogens among different age groups (χ2=83.67, P<0.01). The positive rate of alveolar lavage fluid group was higher than that of deep sputum fluid group [81.6% (614/752) vs. 69.3% (1 137/1 640), χ 2=39.89, P<0.01]. The length of hospital stay of children infected with different pathogens was significantly different (all P<0.01). There was significant difference in duration of fever among children infected with different pathogens (χ2=228.69,103.56, 3.96, 27.38,24.50,41.66, all P<0.05). There were 63 (7.7%) cases of atelectasis, 260 (31.9%) cases of pleurisy and 120 (14.7%) cases of pleural effusion in MP children. Children with Sma were most likely to involve the heart system (2/9), and children with Eco infection had a higher incidence of complications such as those of blood (3/19), urinary (2/19), digestive systems(4/19), systemic inflammatory response syndrome and sepsis (1/19). Conclusions:The main bacterial pathogens of respiratory tract infection in children in Tianjin were MP, Sp, MRSA and Hi. It is suggested that clinicians should not only pay attention to the respiratory symptoms of children, but also pay attention to the complications caused by bacterial pathogen infection, so as to prevent the deterioration of the disease and improve the prognosis.

Objective To investigate the effect and mechanism of interfering the nicotinamide mononucleotide adenylyltransferase 1(NMNAT1)gene in Parkinson's disease(PD)mouse models. Methods Thirty mice were randomly assigned to three groups: the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)group, the small interfering nicotinamide mononucleotide adenylyltransferase 1 (siNMNAT1)+MPTP group, and the control group, with 10 mice in each group.After injecting siRNA-green fluorescent protein(GFP)lentivirals directly into substantia nigra(SN),mice received intraperitoneal injections of MPTP, which was the siNMNAT1 +MPTP group.While the MPTP group was only with injections of MPTP,and the control group was with neither siRNA nor MPTP.Then we assessed the motor coordination ability firstly.To observe the variation of nigrostriatal pathway, the counts of dopamanergic neurons in SN were measured by tyrosine hydroxylase(TH)immunofluorescence staining.And the expression of TH in striatum, which was used to estimate the dopaminergic neurons axonal variation, was analyzed by RT-PCR.Then the expression of TH, SOD1, Bcl2, Bax, Bcl2/Bax in SN was estimated through Western blotting.Results Compared with the control group,the siNMNAT1+MPTP group and the MPTP group decreased significantly in motor coordination ability(creep down time: siNMNAT1 +MPTP group(62.8 ±15.7)s,MPTP group(77.9 ±13.5)s, control group(122.0 ±25.2)s), dopamanergic neuron counts(siNMNAT1 +MPTP group 45.0 ±6.7, MPTP group 68.0 ±11.3, control group 93.0 ± 12.8)and the striatal TH expression(Creep down time: t=-6.291, P=0.000; t=-4.865, P=0.000.Dopamanergic neuron counts:t=-10.482,P=0.000;t=-4.624, P=0.000.TH expression:t=-9.117,P=0.000;t=-5.716, P=0.000).Although the siNMNAT1+MPTP group showed lower coordination ability than the MPTP group, there was no statistically significant difference.Whereas the counts of dopamanergic neurons in SN(t=-5.487, P=0.000), the expression of TH in striatum(t=-5.146,P=0.003),SOD1(t=-4.143, P=0.001)and Bcl2/Bax(t=-6.303, P=0.000)were obviously decreased in the siNMNAT1+MPTP group,in which Bax increased significantly(t=3.550,P=0.002).Conclusions Interfering the expression of NMNAT1 aggravated the neurodegeneration in PD, and the mechanism might be related to oxidative stress and programmed cell death.

Background and objective Tumor-associated fibroblasts (TAF) is an important part of TME, which inhibits the function of immune cells. CD8+ T cells play a significant role in tumor immunity. T-cell membrane possesses a distinct type of molecule with a negative regulatory function. Upon interaction with its corresponding ligand [programmed death factor ligand 1 (PD-L1)], programmed death factor 1 (PD-1) is activated and thus inhibits the kinase activity of T cells. This study aims to explore the possible effects of TAF on PD-L1 expression in lung cancer cells. Methods Lung cancer cell lines H1975 and H520 were co-cultured with (experiment) or without TAF (control) via Transwell assay for through 48 hours under the same culture condition. H1975 and H520 cells were counted using a microscope. The protein and mRNA expression levels of PD-L1 were detected by FCM assay and PCR analysis, respectively. Results The numbers of lung cancer cells in 100μm2 for H1975 and H520 cells are (46±21) and (38±10) in the experiment group, respectively, and (16±5) and (12±5) in the control group, respectively (P<0.05). The expression levels of the PD-L1 protein in H1975 and H520 cells are (20.93%±3.54%) and (19.26%±3.04%) in the experiment group, respectively, and (12.58%±2.52%) and (11.60%±2.65%) in the control group, respectively (P<0.05). The mRNA expression levels in H1975 and H520 cells are (16.45±1.25) and (15.38±2.02) pg/mL in the experiment group, respectively, and (7.78±1.27) and (7.20±1.58) pg/mL (P<0.05) in the control group, respectively (P<0.05). Conclusion TAF promotes the growth and increases the expression of PD-L1 in H1975 and H520 cells.

Purpose To explore the expression of monocarboxylate transporters 4 (MCT4) and its clinicopathologic significance in cer-vical squamous cell carcinoma. Methods The expression level of MCT4 in 72 cervical squamous cell carcinoma tissues and 48 cervi-cal normal tissues were detected by immunohistochemistry assay, and its relationship with clinical pathological features was analyzed. Results The positive rate of MCT4 was 81. 9% (59/72) in cervical squamous cell carcinoma tissues and 35. 4% (17/48) in normal cervical tissues. The positive rate of MCT4 in cervical squamous cell carcinoma was significantly higher than that in cervical normal tis-sues (χ2 =26. 848, P<0. 001). In addition, the expression of MCT4 protein was correlated with clinical stage (χ2 =5. 389, P=0. 020) and lymph node metastasis (χ2 =4. 706, P=0. 030). However, it did not correlated with age (χ2 =1. 238, P=0. 266), tumor differentiation (χ2 =0. 530, P=0. 467) or infiltration degree (χ2 =1. 300, P=0. 254) in patients with cervical squamous cell carcinoma. Conclusion The expression of MCT4 is up-regulated in cervical squamous cell carcinoma and correlate with clinical stage and lymph node metastasis of cervical squamous cell carcinoma. MCT4 may be a biomarker for evaluating the biological behavior of cer-vical squamous cell carcinoma.

OBJECTIVETo investigate the effect of CD59 gene inhibition mediated by RNA interference on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) GLC-P cells in vitro.

METHODSRecombinant plasmids for RNA interference of CD59 gene were constructed and transfected into GLC-P cells via lipofectamine 2000. The stably transfected cells were examined with real-time RT-PCR, MTT assay and enzyme-linked immunosorbent assay to investigate the changes in cell proliferation and apoptosis.

RESULTSCompared with the control cells, the cells transfected with CD59-siRNA showed significantly decreased expression levels of CD59 mRNA (P<0.05) and significantly inhibited cell proliferation.

CONCLUSIONCD59 gene is highly expressed in NSCLC and RNA interference-mediated CD59 silencing can strongly inhibit the proliferation and induce apoptosis in GLC-P cells, which shed light on a potentially new target for targeted gene therapy of NSCLC.

Apoptosis ; CD59 Antigens ; genetics ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Genetic Therapy ; Humans ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Transfection

Objective To investigate the effect of CD59 gene inhibition mediated by RNA interference on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) GLC-P cells in vitro. Methods Recombinant plasmids for RNA interference of CD59 gene were constructed and transfected into GLC-P cells via lipofectamine 2000. The stably transfected cells were examined with real-time RT-PCR, MTT assay and enzyme-linked immunosorbent assay to investigate the changes in cell proliferation and apoptosis. Results Compared with the control cells, the cells transfected with CD59-siRNA showed significantly decreased expression levels of CD59 mRNA (P<0.05) and significantly inhibited cell proliferation. Conclusions CD59 gene is highly expressed in NSCLC and RNA interference-mediated CD59 silencing can strongly inhibit the proliferation and induce apoptosis in GLC-P cells, which shed light on a potentially new target for targeted gene therapy of NSCLC.

Objective To investigate the effect of CD59 gene inhibition mediated by RNA interference on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) GLC-P cells in vitro. Methods Recombinant plasmids for RNA interference of CD59 gene were constructed and transfected into GLC-P cells via lipofectamine 2000. The stably transfected cells were examined with real-time RT-PCR, MTT assay and enzyme-linked immunosorbent assay to investigate the changes in cell proliferation and apoptosis. Results Compared with the control cells, the cells transfected with CD59-siRNA showed significantly decreased expression levels of CD59 mRNA (P<0.05) and significantly inhibited cell proliferation. Conclusions CD59 gene is highly expressed in NSCLC and RNA interference-mediated CD59 silencing can strongly inhibit the proliferation and induce apoptosis in GLC-P cells, which shed light on a potentially new target for targeted gene therapy of NSCLC.