Objective:To discuss the promotion effect of chemokine C-C motif ligand 19(CCL19)induced macrophage M1 polarization on chronic pancreatitis of the mice,and to clarify its related mechanism.Methods:Ten male C57BL/6N mice were selected,and the pancreatic acinar cells and peritoneal macrophages were extracted from these mice to construct the macrophage-acinar cell co-culture system.The co-culture system cells were divided into control group,model group,and small interfering RNA CCL19(si-CCL19)group.The morphology of the acinar cells in various groups were observed under microscope.Forty mice were randomly selected and divided into normal group and chronic pancreatitis group,and there were 20 mice in each group.HE staining was used to observe the pathomorphology of pancreatic tissue of the mice in two groups;immunofluorescence staining was used to observe the expressions of cytokeratin 19(CK19),amylase,M1 macrophage-related markers inducible nitric oxide synthase(iNOS),and F4/80 in pancreatic tissue of the mice in two groups and morphology of follicular cells and the expressions of CK19,amylase in the co-culture system cells in various groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β in serum of the mice in two groups and in the co-culture system cells in various groups;immunohistochemistry was used to observe the expression of CCL19 protein in pancreatic tissue of the mice in two groups;Western blotting method was used to detect the expression levels of CCL19 protein and two nuclear factor-κB(NF-κB)signaling pathway-related proteins P65,phosphorylate P65(p-P65),kappa B inhibitor of kinase α/β(IKKα/β),phosphorylated IKKα/β(p-IKKα/β),IkBα,phosphorylated IκBα(p-IκBα)in pancreatic tissue of the mice in two groups and in the co-culture system cells in various groups.Results:The HE staining results showed that the acinar cells in pancreatic tissue of the mice in normal group were tightly arranged;compared with normal group,the acinar cells of the mice in chronic pancreatitis group showed obvious vacuolation and acinar cell ductal metaplasia,indicating successful preparation of the mouse pancreatitis model.The immunofluorescence staining results showed that compared with control group,the acinar cells in model group exhibited severe vacuolation,the CK19 expression was significantly increased,and the amylase expression was significantly decreased;compared with model group,the acinar cell ductal metaplasia in si-CCL19 group was decreased,the CK19 expression was significantly decreased,and the amylase expression was significantly increased;compared with normal group,the expression of amylase in pancreatic tissue of the mice in chronic pancreatitis group was significantly decreased,while the expressions of CK19 and M1 macrophage markers iNOS and F4/80 were significantly increased.The ELISA results showed that compared with normal group,the serum levels of TNF-α,IL-6,and IL-1β of the mice in chronic pancreatitis group were significantly increased(P<0.05);compared with control group,the levels of TNF-α,IL-6,and IL-1β in the cells in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α,IL-6,and IL-1β in the cells in si-CCL19 group were significantly decreased(P<0.05).The immunohistochemistry results showed that compared with normal group,the expression of CCL19 protein in pancreatic tissue of the mice in chronic pancreatitis group was significantly increased.The Western blotting results showed that compared with normal group,the expression levels of CCL19 protein and NF-κB signaling pathway-related proteins p-P65,p-IKKα/β,and p-IκBα in pancreatic tissue of the mice in chronic pancreatitis group were significantly increased(P<0.05);compared with control group,the expression levels of CCL19,p-IKKα/β,p-P65,and p-IκBα proteins in the cells in model group were significantly increased(P<0.05);compared with model group,the expression levels of CCL19,p-IKKα/β,p-P65,and p-IκBα proteins in the cells in si-CCL19 group were decreased(P<0.05).Conclusion:CCL19 promotes the macrophage M1 polarization through the NF-κB signaling pathway,induces the formation of inflammatory microenvironment,and promotes the occurrence and development of pancreatitis.

Objective:To analyze the application value of noninvasive prenatal testing (NIPT) in screening fetal chromosome aneuploidy in cautious population.Methods:The data of 19 590 pregnant women who voluntarily received NIPT in the Department of Gynecology and Obstetrics, General Hospital of Tianjin Medical University from October 2016 to June 2020 were retrospectively analyzed. The patients were divided into groups according to age, type of pregnancy and number of fetuses. The screening efficiency of NIPT was compared among different groups. Meanwhile, the screening efficacy of second trimester serum screening (STSS), STSS combined with NIPT and direct NIPT screening was compared and analyzed.Results:The positive predictive values of NIPT for trisomy 21, trisomy 18 and trisomy 13 were 87.18%, 54.84% and 41.18%, respectively. In pregnant women with advanced age (≥35 years old), in vitro fertilization and embryo transfer (IVF-ET) and twin pregnancy, NIPT sensitivity was all 100%, specificity was 99.79%, 99.82% and 100%, positive predictive value was 85.25%, 50.00% and 100%, respectively. The positive predictive value of ≥35 years old patients was significantly higher than that of <35 years old patients (61.54%, P=0.003), and the other indicators were similar to those of the young (<35 years old), conceived naturally and singleton group (all P>0.05). Compared with STSS, combined screening and direct NIPT screening had higher positive predictive value (1.20%, 68.75%, 73.74%; all P<0.001) and lower false positive rate (27.83%, 0.19%, 0.17%; all P<0.001). Conclusion:NIPT has high sensitivity, specificity and positive predictive value for fetal chromosome aneuploidy screening in pregnant women with advanced age, STSS high risk, IVF-ET and twin pregnancy, which greatly relieves the pressure of prenatal diagnosis and reduces the birth rates of children with chromosome disease.

Objective:To analyze the application value of noninvasive prenatal testing (NIPT) in screening fetal chromosome aneuploidy in cautious population.Methods:The data of 19 590 pregnant women who voluntarily received NIPT in the Department of Gynecology and Obstetrics, General Hospital of Tianjin Medical University from October 2016 to June 2020 were retrospectively analyzed. The patients were divided into groups according to age, type of pregnancy and number of fetuses. The screening efficiency of NIPT was compared among different groups. Meanwhile, the screening efficacy of second trimester serum screening (STSS), STSS combined with NIPT and direct NIPT screening was compared and analyzed.Results:The positive predictive values of NIPT for trisomy 21, trisomy 18 and trisomy 13 were 87.18%, 54.84% and 41.18%, respectively. In pregnant women with advanced age (≥35 years old), in vitro fertilization and embryo transfer (IVF-ET) and twin pregnancy, NIPT sensitivity was all 100%, specificity was 99.79%, 99.82% and 100%, positive predictive value was 85.25%, 50.00% and 100%, respectively. The positive predictive value of ≥35 years old patients was significantly higher than that of <35 years old patients (61.54%, P=0.003), and the other indicators were similar to those of the young (<35 years old), conceived naturally and singleton group (all P>0.05). Compared with STSS, combined screening and direct NIPT screening had higher positive predictive value (1.20%, 68.75%, 73.74%; all P<0.001) and lower false positive rate (27.83%, 0.19%, 0.17%; all P<0.001). Conclusion:NIPT has high sensitivity, specificity and positive predictive value for fetal chromosome aneuploidy screening in pregnant women with advanced age, STSS high risk, IVF-ET and twin pregnancy, which greatly relieves the pressure of prenatal diagnosis and reduces the birth rates of children with chromosome disease.

Objective To observe the effect of nicotine(NIC) on the activation and resultant death of microglia induced by LPS. Methods The animal model that exposed to chronic nicotine treatment was established and LPS was injected intraperitoneally to induce the activation of microglia. Furthermore, the CD11b-positive microglia in cerebral cortex, hippocampal and substantia ngra were observed through immunohistochemical staining. BV2 cells(Microglial cell line of mouse) were subcultured, simultaneously the following kits were used including CCK-8 kit assay for cell activity, Nitric oxide assay kit assay for NO release, RT-PCR assay for the iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, Western blotting assay for the protein expression of P-I-κB and Caspase-3. Results Nicotine suppressed the CD11b-positive microglia expression in cerebral cortex,hippocampal and substantia ngra induced by LPS;Nicotine inhibited the activation-induced cell death (AICD), attenuated NO release, reduced iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, decreased the protein expression including P-I-κB and Caspase-3 of BV2 cells. Conclusion Nicotine pretreatment can suppress the activation and resultant death of microglial cells induced by LPS, which suggests that nicotine may play a neuroprotective role on inflammatory reaction of brain.;

Objective To explore the effect of nicotine on the degeneration of dopaminergic neurons in PD rats. Methods Forty-five SD rats were randomly divided into three groups: PBS group (CON), normal saline + LPS ( NS ) group and nicotine +LPS ( NIC ) group. On 24 hours after LPS or PBS injection, inducible nitric oxide synthase ( iNOS ) protein expression was examined by immunoblotting. ON 14d after LPS or PBS injection, the numbers of tyrosine hydroxylase ( TH ) positive neurons and morphological changes of OX-42 positive cells in the substantia nigra(SN) were observed by immunohistochemistry. TH mRNA and TH protein expressions were examined by RT-PCR or immunoblotting. Results Compared with CON group, iNOS protein increased markedly 24 hours after LPS injection in NS group. TH positive neurons, TH mRNA and TH protein in the SN decreased remarkably 14 days after nigrainjiection. Most of microlglial showed big cell body with thicker and shorter processes. However, nicotine reversed the above changes, Compared with NS group, TH positive neurons, TH mRNA and TH protein in the SN increased remarkably in NIC. Besides, most microglia showed small cell body with slim and long processes. Conclusion Nicotine could prevent LPS-induced degeneration of DA neurons, probably because of that the pretreatment with nicotine blocks the activation of microglia and the expression iNOS protein.

Objective Our aim was to observe the effects of prenatal nicotine exposure on development of cerebral cortex and hippocampus in the offsprings of pregnant mice and expressions of NCAM proteins. Methods We established prenatally nicotine exposed(PNE) animal models.We investigated the offspring's structural changes of cerebral cortex and hippocampus with HE staining.Using immunohistochemistry and Western blotting we observed the changes of expressions of NCAM proteins in the neonatal mice. Results Cerebral cortex and hippocampus as well as hippocampal cell layers of the PNE offsprings were thinner than that of the control group.The expressions of PSA-NCAM in cerebral cortex and hippocampus of the PNE offsprings were higher than the control group while the expressions of NCAM-180,140 and 120 weaker.Conclusion Prenatally nicotine exposure can prevent normal morphogenesis of cerebral cortex and hippocampus of mice and can change the expressions of NCAM proteins in cerebral cortex and hippocampus of neonatal mice.

Objective To investigate the effects of acute nicotine exposure on expression of Caspase-3 and apoptosis of neuron in adolescent rat's brain. Methods The Caspase-3 expression was detected with immunohistochemistry and Western blotting. Apoptosis cells were detected by TUNEL method. Results The Caspase-3 expression in cerebral cortex, hippocampal dentate gyrus, CA2 and CA3 of the experimental group was higher than that of the control group through immunohistiochemistry. The expression of Caspase-3 prototype and active fragments in cerebral cortex and hippocampus of the experimental group was higher than that of the control group with Western blotting. The TUNEL positive cells in cerebral cortex and hippocampal dentate gyrus of the experimental group were more than that of the control group.Conclusion Acute nicotine exposure can activate Caspase-3 and increase neuron apoptosis in cerebral cortex and hippocampal dentate gyrus of the adolescent rat's brain.

Objective To observe the regulation of ?-7 neuronal nicotinic acetylcholine receptors(?-7-nAChRs) by nicotine in the ventral tegmental area(VTA),the substantia nigra(SN) and the cortex of the rat. Methods Nicotine exposed animal models were established.The changes of ?-7-nAChRs proteins and mRNA were observed by immunohistochemistry and in situ hybridization in VTA,SN and cortex,and the expression of ?-7-nAChRs proteins by Western blotting in PC12 cells. Results The expression of ?-7-nAChRs proteins in cortex and mRNA in VTA and SN were upregulated by nicotine.The expressions of ?-7-nAChRs in PC12 cells were in proportion to and dependent on the time for nicotine to function and the dose.Conclusion Both ?-7-nAChRs mRNA in VTA and SN in transcriptional mechanism and ?-7-nAChRs protein in the cortex at the translation level were upregulated.