Objective To explore how Vibrio vulnificus (Vv) invades dendritic cells (DC) and induces acute necrosis of DC via toll-like receptor (TLR) 2 and 4 pathways.Methods Vv 1.1758 strain and DC 2.4 mixed culture model was established,observed the infection rates of DC with optical microscope,the location of Vv and structural changes of DC by transmission electron microscope.The expression levels of TLR2 and TLR4 mRNA were determined by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and tumor necrosis factor-α (TNFα) protein titers were measured by enzyme-linked immunosorbent assay (ELISA).DNA ladder qualitative test was used to detect cell apoptosis,while flow cytometry was used to quantify cell apoptosis and necrosis rates.Statistical analysis was done by chi-square test and one-way ANOVE.Results The infection rates of DC after 0.5,1,2,4 and 6 h of mixed culture were (7.8±0.8) ％,(13.9± 1.1) ％,(34.6±4.9) ％,(77.8± 10.2)％ and (95.8 ± 13.1)％,respectively.Vv was generally located in the internal cell membrane of DC 2.4.After 2 h co-culture,nuclear chromatins of DC became active and intranuclear apoptosis bodies appeared.After 4 h,cytoplasmic vacuoles appeared,chromatin gathered,and cell membranes were seriously damaged.After 6 h,mitochondria was highly swelled and distorted,and cell apoptosis and necrosis occurred.TLR2 and TLR4 mRNA levels reached peak values after co-culture for 0.5 h; TNF-α level began to increase at 1 h (P＜0.05) and reached peak values at 2 h.DNA Ladder electrophoresis presented scouring necrosis after 2 h culture and apoptotic bands appeared between 720 bp and 900 bp after 4 to 5 h culture.Early apoptosis rates of DC after 2,4 and 6 h culture were (3.1±3.8)％,(7.8±4.7)％ and (12.7±8.2)％,and necrosis rates of DC were (16.7±12.5)％,(41.6±25.9)％ and (75.5±33.6)％,higher than that of control group (all P＜0.05).Conclusions Vv infects DC and induce DNA degradation through up-regulated expression of TLR2 and TLR4 and increasing of TNF-α inflammatory mediators.During cell degradation,apoptosis and necrosis coexist,while necrosis is predominant.

Objective To investigate the invasion,internalization and the organelles damage of the cultured dendritic cells ( DC2.4 strain) during Vibrio vulnificus (Vv) infection.Methods The study model was the cultured DCs infected by Vv 1.1758 strain.Electron microscopy was used to observe the localization of bacteria in different time point of infection,cell morphology and the process of organelles changes.The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined by the fluorescence microscope.Results The Vv were pinocytosed into the DC cells through double-sides,and localized at 1-2 μm of the inner side membrane.It cost 1.27,1.87,3.43 hours reaching the infection ratio of 25％,50％,75％,respectively.Using electron microscopy,the DCs had been observed the phagosome formation within 1h,chromatin activation within 2 h,chromatin aggregation 4 h,and the significant cytoskeleton structure disruption within 6 h.Endoplasmic reticulum,mitochondria and lysosomes became swollen.In DCs,the protruding filaments gradually reduced,and their shape changed from the point-like to the linearlike aggregation at the inner side of the plasma membrane,extended microtubules disappeared,the microtubules at the outside nuclear membrane striking rearranged.Conclusion After DC was infected by Vv,the bacteria were pinocytosed into the inner side of DC membrane,and the microfilaments were observed to move from the cytoplasm to cell membrane.In addition,the microtubules moved from the synapse and the cell membrane to the nuclear membrane.The high lethality of Vv could provoke to the DCs cytoskeleton rearrangements.

Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To study the application effect of Suhexiang after PCI in the patients with mild cognitive impairment(MCI) complicating acute ST-elevation myocardial infarction.Methods A total of 100 inpatients with MCI complicating acute ST-elevation myocardial infarction in Shijiazhuang People's Hospital were selected and divided into the control group and Suhexiang group according to the random number table method,50 cases in each group.The control group was given the clinical routine treatment of coronary heart disease PCI,and the Suhexiang group was added with Suhexiang 1 g once daily on the basis of the control group,and the course of treatment was 1 month.Each patient conducted the coronary arterial SYNTAXⅡ sco-ring.Resting D-SPECT+regadenoson loaded D-SPECT was used to evaluate the number of myocardial ische-mia total segments,left ventricular ejection fraction,total myocardial perfusion score on 7 d and in 3 months after PCI,MMSE and MoCA were used to evaluate the cognitive impairment on 7 d and in 3 months after PCI,and the adverse drug reactions occurrence in 3 months after PCI was recorded.Results There was no statistically significant difference in MMSE,MoCA,left ventricular ejection fraction,number of myocardial is-chemia segments,and myocardial perfusion total score on 7 d after PCI between the two groups (P>0.05). The MMSE,MoCA,left ventricular ejection fraction,number of myocardial ischemia segments and total myo-cardial perfusion score in 3 months after PCI in the Suhexiang group were significantly better than those in the control group,and the difference was statistically significant (P<0.05).There was no statistically significant difference in adverse reactions between the two groups (P>0.05).Conclusion Suhexiang could significantly improve the myocardial perfusion,cardiac function and cognitive function after PCI in the patients with MCI complicating acute ST-elevation myocardial infarction,but does not increase adverse reactions.

Objective:To analysis the income and expenditure of HIV-positive pregnant women and the catastrophic health expenditure of their households in high AIDS endemic areas in Liangshan prefecture.Methods:From December 2018 to January 2019, a total of 250 pregnant women were recruited from 2 of 17 counties in Liangshan Prefecture, including 133 HIV-positive pregnant women and 117 HIV-negative pregnant women. The data, including basic information of pregnant women, basic information of the family, annual family income in 2017, annual family health expenditure in 2017, and maternal and child-related expenditure in 2017, were collected for analyzing the incidence of catastrophic health expenditure in the family.Results:The average annual income and average annual health expenditure of HIV-positive pregnant women households were 7 000 CNY and 2 000 CNY, while those of HIV-negative pregnant women households were 10 000 CNY and 3 000 CNY, respectively. Based on the criteria of 15%, 25% and 40%, the incidence of catastrophic health expenditure of HIV-positive pregnant households was 39.10%, 33.83% and 27.82%, with average differences of 34.84%, 31.17% and 26.65%, respectively, while that of HIV-negative pregnant women households was 38.46%, 33.33% and 23.93%, with average differences of 31.68%, 28.35% and 24.22%, respectively.Conclusion:The incidence of catastrophic health expenditure of pregnant households in high AIDS endemic areas in Liangshan prefecture is high. The incidence of HIV-positive families is slightly higher than that of HIV-negative families. We suggest that medical insurance compensation in Liangshan prefecture should be improved to reduce the impact of catastrophic health expenditure.

Objective:To analysis the income and expenditure of HIV-positive pregnant women and the catastrophic health expenditure of their households in high AIDS endemic areas in Liangshan prefecture.Methods:From December 2018 to January 2019, a total of 250 pregnant women were recruited from 2 of 17 counties in Liangshan Prefecture, including 133 HIV-positive pregnant women and 117 HIV-negative pregnant women. The data, including basic information of pregnant women, basic information of the family, annual family income in 2017, annual family health expenditure in 2017, and maternal and child-related expenditure in 2017, were collected for analyzing the incidence of catastrophic health expenditure in the family.Results:The average annual income and average annual health expenditure of HIV-positive pregnant women households were 7 000 CNY and 2 000 CNY, while those of HIV-negative pregnant women households were 10 000 CNY and 3 000 CNY, respectively. Based on the criteria of 15%, 25% and 40%, the incidence of catastrophic health expenditure of HIV-positive pregnant households was 39.10%, 33.83% and 27.82%, with average differences of 34.84%, 31.17% and 26.65%, respectively, while that of HIV-negative pregnant women households was 38.46%, 33.33% and 23.93%, with average differences of 31.68%, 28.35% and 24.22%, respectively.Conclusion:The incidence of catastrophic health expenditure of pregnant households in high AIDS endemic areas in Liangshan prefecture is high. The incidence of HIV-positive families is slightly higher than that of HIV-negative families. We suggest that medical insurance compensation in Liangshan prefecture should be improved to reduce the impact of catastrophic health expenditure.

Objective To discuss the clinical safety,feasibility and efficacy of transcatheter arterial infusion chemotherapy(TAI)combined with lipiodol chemoembolization in the treatment of advanced colorectal cancer(CRC).Methods The clinical data of 37 patients with advanced CRC,who received TAI combined with lipiodol chemoembolization at the First Affiliated Hospital of Zhengzhou University of China between June 2016 and December 2022,were retrospectively analyzed.The clinical efficacy was evaluated,the progression-free survival(PFS)and the serious complications were recorded.Results A total of 55 times of TAI combined with lipiodol chemoembolization procedures were successfully accomplished in the 37 patients.The mean used amount of lipiodol emulsion was 2.9 mL(0.8-10 mL).No serious complications such as bleeding and intestinal perforation occurred.The median follow-up time was 24 months(range of 3-48 months).The postoperative one-month,3-month,6-month and 12-month objective remission rates(ORR)were 67.6%(25/37),67.6%(25/37),64.9%(24/37)and 56.8%(21/37)respectively,and the postoperative one-month,3-month,6-month and 12-month disease control rates(DCR)were 91.9%(34/37),91.9%(34/37),89.2%(33/37)and 81.1%(30/37)respectively.The median PFS was 16 months(range of 2-47 months).As of the last follow-up,22 patients survived and 15 patients died of terminal stage of tumor.Conclusion Preliminary results of this study indicate that TAI combined with lipiodol chemoembolization is clinically safe and effective for advanced CRC,and it provide a new therapeutic method for patients with advanced CRC.