Objective:To investigate the influences of long non-coding RNA metastasis-associated lung adenocarcinoma tran-script 1(lncRNA MALAT1)/miR-876-5p/forkhead box protein M1(FOXM1)axis on TNF-α-induced proliferation,apoptosis and in-flammatory response of HaCaT cells.Methods:HaCaT cells were grouped into Ct group,Model group,si-NC group,si-MALAT1 group,mimic NC group,miR-876-5p mimic group,si-MALAT1+inhibitor NC group,and si-MALAT1+miR-876-5p inhibitor group.Except for the Ct group,cells in other groups were treated with 25 μg/L TNF-α to induce the in vitro cell model of psoriasis,and after 24 hours of TNF-α induction,the corresponding transfectants were transfected for 48 hours for subsequent experiments.qRT-PCR was applied to detect the expression of MALAT1 and miR-876-5p in cells;CCK-8 method and EdU staining were applied to detect cell pro-liferation;flow cytometry was applied to detect apoptosis;ELISA method was applied to detect the levels of IL-6,TNF-α,and IL-1β in cell supernatant;Western blot was applied to detect the protein expressions of FOXM1,proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax)and B-lymphocytoma-2(Bcl-2);dual-luciferase reporter gene assay was applied to verify the rela-tionship between MALAT1 and miR-876-5p,miR-876-5p and FOXM1;and RNA pull down experiments were applied to verify the re-lationship between MALAT1 and miR-876-5p.Results:Compared with the control group,the expressions of MALAT1 and FOXM1 protein expression in the experimental group were increased,and the expression of miR-876-5p was decreased(P<0.05);compared with Ct group,the expressions of MALAT1 and FOXM1 protein expression in HaCaT cells,A450 value,EdU positive rate,the levels of IL-6,TNF-α,IL-1β,and the protein expressions of PCNA and Bcl-2 in cell supernatant in Model group increased,the expression of miR-876-5p,apoptosis rate and the protein expression of Bax were decreased(P<0.05);silencing MALAT1 or overexpressing miR-876-5p could inhibit the proliferation and inflammatory response of HaCaT cells induced by TNF-α,and promote cell apoptosis;miR-876-5p inhibitor attenuated the inhibitory effects of silencing MALAT1 on TNF-α-induced HaCaT cell proliferation and inflammatory response,and the promotion on cell apoptosis;MALAT1 targeted and regulated the miR-876-5p/FOXM1 axis.Conclusion:Silencing MALAT1 may inhibit the expression of FOXM1 by up-regulating miR-876-5p,thereby inhibiting the proliferation and inflammatory re-sponse of HaCaT cells induced by TNF-α,and promoting cell apoptosis.

Objective:To investigate the influences of long non-coding RNA metastasis-associated lung adenocarcinoma tran-script 1(lncRNA MALAT1)/miR-876-5p/forkhead box protein M1(FOXM1)axis on TNF-α-induced proliferation,apoptosis and in-flammatory response of HaCaT cells.Methods:HaCaT cells were grouped into Ct group,Model group,si-NC group,si-MALAT1 group,mimic NC group,miR-876-5p mimic group,si-MALAT1+inhibitor NC group,and si-MALAT1+miR-876-5p inhibitor group.Except for the Ct group,cells in other groups were treated with 25 μg/L TNF-α to induce the in vitro cell model of psoriasis,and after 24 hours of TNF-α induction,the corresponding transfectants were transfected for 48 hours for subsequent experiments.qRT-PCR was applied to detect the expression of MALAT1 and miR-876-5p in cells;CCK-8 method and EdU staining were applied to detect cell pro-liferation;flow cytometry was applied to detect apoptosis;ELISA method was applied to detect the levels of IL-6,TNF-α,and IL-1β in cell supernatant;Western blot was applied to detect the protein expressions of FOXM1,proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax)and B-lymphocytoma-2(Bcl-2);dual-luciferase reporter gene assay was applied to verify the rela-tionship between MALAT1 and miR-876-5p,miR-876-5p and FOXM1;and RNA pull down experiments were applied to verify the re-lationship between MALAT1 and miR-876-5p.Results:Compared with the control group,the expressions of MALAT1 and FOXM1 protein expression in the experimental group were increased,and the expression of miR-876-5p was decreased(P<0.05);compared with Ct group,the expressions of MALAT1 and FOXM1 protein expression in HaCaT cells,A450 value,EdU positive rate,the levels of IL-6,TNF-α,IL-1β,and the protein expressions of PCNA and Bcl-2 in cell supernatant in Model group increased,the expression of miR-876-5p,apoptosis rate and the protein expression of Bax were decreased(P<0.05);silencing MALAT1 or overexpressing miR-876-5p could inhibit the proliferation and inflammatory response of HaCaT cells induced by TNF-α,and promote cell apoptosis;miR-876-5p inhibitor attenuated the inhibitory effects of silencing MALAT1 on TNF-α-induced HaCaT cell proliferation and inflammatory response,and the promotion on cell apoptosis;MALAT1 targeted and regulated the miR-876-5p/FOXM1 axis.Conclusion:Silencing MALAT1 may inhibit the expression of FOXM1 by up-regulating miR-876-5p,thereby inhibiting the proliferation and inflammatory re-sponse of HaCaT cells induced by TNF-α,and promoting cell apoptosis.

Objective:To explore effect of miR-125a-3p on skin injury and inflammatory response in psoriasis rats and its mechanism.Methods:SD rats were randomly divided into control group,psoriasis group,miR-NC group and miR-125a-3p group.Psoriasis Area and Severity Index(PASI)score and Baker score were measured on rats;levels of IL-6,IL-1β and TNF-α in rat skin tissue were detected by ELISA;qRT-PCR was used to detect miR-125a-3p expression;mRNA and protein expressions of forkhead box protein M1(FOXM1)in rat skin tissue were detected by qRT-PCR and Western blot.Keratinocytes from psoriasis rats were isolated and cultured,targeting relationship between miR-125a-3p and FOXM1 was verified by dual-luciferase reporter gene assay.Inhibiting or overexpressing miR-125a-3p and FOXM1 was overexpressed on basis of overexpressing miR-125a-3p,respectively.miR-125a-3p,FOXM1 mRNA and protein expressions in cells and IL-6,IL-1β,TNF-α levels in cell culture supernatants were detected,CCK-8 method was applied to detect cell viability,and flow cytometry was used to detect apoptosis rate.Results:Compared with control group,miR-125a-3p expression in skin tissue of rats in psoriasis group was decreased,PASI score,Baker score,IL-6,IL-1β,TNF-α levels,and FOXM1 mRNA and protein expressions were increased(P<0.05);compared with psoriasis group and miR-NC group,expression of miR-125a-3p in skin tissue of rats in miR-125a-3p group was increased,PASI score,Baker score,IL-6,IL-1β,TNF-α levels,and expressions of FOXM1 mRNA and protein were decreased(P<0.05).There was a targeting relationship between miR-125a-3p and FOXM1.After inhibiting miR-125a-3p expression,FOXM1 mRNA and protein expressions in cells,cell viability and IL-6,IL-1β,TNF-α levels in cell culture supernatant were increased,and apoptosis rate was decreased(P<0.05),while overexpression of miR-125a-3p had opposite effect.Overexpression of FOXM1 attenuated effects of overexpression of miR-125a-3p on cell viability,apopto-sis rate and inflammatory response.Conclusion:miR-125a-3p is lowly expressed in skin lesions of psoriasis rats,whose overexpression may inhibit proliferation of keratinocytes and promote apoptosis by targeting FOXM1,improve skin injury and reduce inflammatory response in psoriasis rats.

Objective To investigate the perioperative analgesic effect of ultrasound-guided bilateral thora-columbar interfascial plane block(TLIPB)in patients with discogenic low back pain(DLBP)undergoing percuta-neous transforaminal endoscopic discectomy(PTED).Methods Fifty-seven patients with discogenic low back pain admitted to the First Affiliated Hospital of Anhui Medical University from January 2022 to April 2022 were randomly divided into group A(control group)with 28 patients and group B(ultrasound-guided bilateral thoracolumbar inter-fascialinterfacial plane block group)with 29 patients.The differences of visual analogue scale(VAS)at rest and turning over were compared between the two groups preoperative(t0),2 hours postoperative(t1),6 hours postop-erative(t2),12 hours postoperative(t3)and 24 hours postoperative(t4).The differences of quality of recovery-15 scores(QoR-15)were compared between the two groups preoperative and 24 hours postoperative.The changes of mean arterial pressure(MAP)and heart rate(HR)were compared between the two groups after entering the oper-ating room(T0),at the time of skin incision(T1),at the time of foraminoplasty(T2),at the time of the most severe pain recognized by the surgeon(T3),and at the end of the operation(T4).Adverse events were recorded during the operation and within 24 hours postoperative.Results All patients successfully completed the operation and ultrasound-guided bilateral TLIPB,without intervertebral space infection,spinal cord,nerve root and vascular injury,and serious complications such as nausea and vomiting.VAS scores at rest and turning over and QoR-15 scores at 24 hours postoperative were significantly lower than preoperative in the two groups(P<0.05).There was no significant difference in VAS scores between the two groups at rest at preoperative and postoperative time points(P>0.05).There were significant differences in VAS scores at 2 hours,6 hours and 12 hours postoperative between the two groups(P<0.05).There were significant difference in QoR-15 scores between the two groups at preoperative and 24 hours postoperative(P>0.05).There was a significant difference in QoR-15 scores between the two groups at 24 hours postoperative(P>0.05).There were significant differences in MAP and HR between the two groups at the time of foraminoplasty(T2)and at the time of the most severe pain recognized by the surgeon(T3)(P<0.05).Conclusions Ultrasound-guided bilateral thoracolumbar interfascial plane block can effectively relieve the pain after PTED,reduce the occurrence of perioperative stress response and adverse events,accelerate the postoperative rehabilitation of patients,and shorten the postoperative duration of hospitalization.

Objective:To explore clinical value of nucleic acid detection for hepatitis B virus (HBV) screening in hospitalized patients.Methods:This cross-sectional study collected and analyzed plasma samples from patients admitted to 10 domestic medical institutions from July 2021 to December 2021. Serological immunoassay and nucleic acid screening were used to simultaneously detect hepatitis B markers such as hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B e Antigen (HBeAg), hepatitis B e antibody (HBeAb), hepatitis B core antibody (HBcAb),and HBV DNA. Statistical analysis was performed on the serology, nucleic acid test results and clinical information of the patients.Results:Of the 8 655 collected samples, HBsAg was positive in 216 (2.50%) samples,HBV DNA was positive in 238 (2.75%) samples ( P>0.05); 210 (2.43%) samples were positive for both HBsAg and HBV DNA, 28 (0.32%) were HBsAg negative and HBV DNA positive, 6 cases (0.07%) were HBsAg positive and HBV DNA negative. Conclusion:These results indicate that the HBV DNA testing is equally effective as hepatitis B virus serological detection for hepatitis B virus screening in hospitalized patients.

Objective:This multi-centre study was conducted to assess the efficacy of various preoperative/pre-transfusion screening methods for blood transmitted disease.Methods:From July 2021 to December 2021, plasma samples of patients admitted to 10 hospitals were collected for screening preoperative/pre-transfusion blood transmitted disease. Nucleic acid detection technology was used to detect hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA and human immunodeficiency virus (HIV)(1+2) RNA, and the results were compared with the immuno-serological methods. χ 2 test and Kappa test were used to analyze the efficacy of these two methods. Results:A total of 8 655 valid specimens were collected from 10 hospitals. There was a statistically significant difference in the positive detection rate of HCV between the two methods ( P<0.001). There was no significant difference in the positive detection rate of HBV and HIV assessed by the two methods ( P>0.05), but the number of positive cases detected by HBV DNA and HIV RNA (218 and 4 cases) was significantly higher than the corresponding serological results (216 and 2 cases). At the same time, there were HBV, HCV and HIV immuno-serological omissions by the immuno-serological methods, among which 28 cases were HBsAg negative and HBV DNA positive, 2 cases were HCV antibody negative and HCV RNA positive, and 2 cases were HIV antigen/antibody negative and HIV RNA positive. In addition, in the 66 samples with inconsistent results from the two detection methods, 83.3% (55/66), 68.2% (45/66), 63.6% (42/66) and 62.1% (41/66) of patients aged was>45 years, tumor, surgery and male, respectively. Conclusions:Compared with immuno-serological tests, nucleic acid tests have the advantage in terms of sensitivity on detecting HBV, HCV and HIV infection and could reduce missed detection. The risk of transmission can be reduced by adding HBV, HCV, and HIV nucleic acid tests to preoperative/pre-transfusion immuno-serological tests screening for patients over 45 years of age and tumor patients.

Objective:To compare the cost-effectiveness of hospitalized Chinese patients undergoing nucleic acid screening strategies for hepatitis B and hepatitis C, immunological screening strategy, and no screening strategy under different willingness to pay (WTP). The results might aid to decision-making for the optimal strategy.Methods:In this study, nucleic acid screening, immunological screening and no screening were used as screening strategies, and China′s GDP in 2021 (80 976 yuan) was used as the threshold of WTP to construct a Markov model. After introducing parameters related to the diagnosis and treatment of hepatitis B and C in inpatients, a cohort population of 100 000 inpatients was simulated by TreeAge Pro 2021 software, the total cost, total health effects, incremental cost-effectiveness ratio and average cost-effectiveness ratio of different screening strategies were calculated, and cost-effectiveness analysis was conducted. Univariate and probabilistic sensitivity analysis were used to assess the impact of parameter uncertainty on the final results.Results:Compared with the non-screening strategy, the incremental total cost of the hepatitis B immunological screening strategy for cohort patients was 11 049 536 yuan, and the incremental cost-effectiveness ratio was 24 762 yuan/quality-adjusted life years (QALY), while the total incremental cost of nucleic acid screening was 19 208 059 yuan, and the incremental cost-effectiveness ratio was 29 873 yuan/QALY; the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 834 yuan/QALY. Compared with the non-screening strategy, the incremental cost-effectiveness ratio of hepatitis C immunological screening strategy was 5 731 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening strategy was 8 722 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 591 yuan/QALY. The results of probabilistic sensitivity analysis showed that when the cost of nucleic acid testing exceeded 214.53 yuan, it was not cost-effective to perform hepatitis B nucleic acid screening under the WTP as 1 fold GDP. When the cost of nucleic acid testing exceeded 132.18 yuan, it was not cost-effective to conduct hepatitis C screening under the WTP as 1 fold GDP.Conclusions:Nucleic acid screening strategy can achieve more cost-effectiveness and is worthy of vigorous promotion. Compared with no screening, both the nucleic acid and immunological screening strategies are cost-effective, and hepatitis nucleic acid screening is the optimal strategy for hospitalized patients.

Objective To explore the risk factors of pulmonary artery hypertension (PAH) and the its relationship with T cell subsets to provide a foundation for the prevention and treatment of PAH.Methods 154 maintained hemodialysis (MHD) patients in our dialysis center were recruited according to the criterion and divided into two groups subsequently:PAH group (pulmonary artery systolic pressure,PASP ＞ 35 mmHg) and non-PAH group (PASP≤35 mmHg).The related clinical,biochemical and ultrasonic cardiogram data were collected and peripheral blood was acquired to detect the expressions of the surface antigen CD3,CD4,CD8 and CD69 with flow cytometry.Logistic regression analysis was used to find out the relationship between PAH and T cell subsets.Results There was no significant difference between 56 cases of PAH and 98 cases of non-PAH as regards gender,age,mean systolic and diastolic pressure,dialysis durations,morbidities of hypertension and diabetes,smoking rate,and left ventricular diameter.Compared with the non-PAH group,the PAH group demonstrated a lower percent of CD8 T cells and CD8 CD69 T cells,but a much higher left atrial diameter (LAD),Interventricular septum thickness,left ventricular posterior wall thickness,and NT-proBNP.The percentage of T cells,CD4 T cells and CD4 CD69 T cells showed no difference between the two groups.Multivariate analysis confirmed that PAH was negatively independently associated with the percentage of CD8 T cells and CD8CD69 T cells.Conclusions The decreased percentage of CD8 T cells and CD8CD69 T cells in the peripheral blood is a risk factor of PAH in maintained hemodialysis patients,and CD8 T cells may play an important role in the genesis of PAH.

Objective To evaluate the effect of hydrogen peroxide (H2O2) on a senescence marker protein-30 (SMP30) and an autophagy-related protein microtubule-associated protein 1 light chain 3 type Ⅱ (LC3-Ⅱ) in normal human skin fibroblasts (NHSFs).Methods NHSFs were isolated from the foreskin of children,and subjected to culture in vitro.The second-to fourth-passage NHSFs were treated with 150 μmol/L H2O2 for 2 hours to establish a model for cellular senescence,while un-treated NHSFs served as control group.Senescence-associated β-galactosidase (SA-β-gal) staining was performed to determine the percentage of senescent cells,indirect immunofluorescence assay to determine the expression of the autophagy-related protein LC3,reverse transcription PCR (RT-PCR) to measure the mRNA expression of SMP30,and Western blot analysis to measure the protein expression of SMP30 and LC3.Results The percentage of senescent (SA-β-gal-positive) cells was significantly higher in the H2O2 group than in the control group (41.70％ ± 2.95％ vs.3.03％ ± 0.25％,t =22.59,P ＜ 0.05).Indirect immunofluorescence assay showed that the percentage of LC3-positive cells was significantly lower in the H2O2 group than in the control group (12.60％ ± 1.57％ vs.23.67％ ± 3.04％,t =5.61,P ＜ 0.05).As Western blot analysis showed,no significant difference in the expression of LC3-Ⅰ (LC3-Ⅰ/glyceraldehyde-3-phosphate dehydrogenase [GAPDH] ratio) was observed between the H2O2 group and control group (0.40 ± 0.02 vs.0.41 ± 0.04,P ＞ 0.05),while the H2O2 group showed significantly lower expression of LC3-Ⅱ (LC3-Ⅱ/GAPDH ratio:0.20 ± 0.02 vs.0.80 ± 0.03,t =29.69,P ＜ 0.05) and lower LC3-Ⅱ/LC3-Ⅰ ratio (0.51 ± 0.03 vs.1.98 ± 0.23,t =10.967,P ＜ 0.05) compared with the control group.Moreover,the mRNA and protein expression of SMP30 (SMP30/GAPDH ratio) was significantly lower in the H2O2 group than in the control group (mRNA:0.16 ± 0.01 vs.0.35 ± 0.01;protein:0.27 ± 0.02 vs.0.63 ± 0.02,both P ＜ 0.05).Conclusion H2O2 can decrease the expression of SMP30 and LC3-Ⅱ in NHSFs,and accelerate the senescence of NHSFs.

Objective: To investigate the possible mechanism related to the protective effects of salvianolate in H9c2 cells underwent hypoxia/reoxygenation (H/R)-injury. Methods: H9c2 cells were divided into four groups: control group, salvianolate group (S group), H/R group, and salvianolate+ H/R group(S+ H/R group), in which the H9c2 cells were pretreated with salvianolate before H/R-treatment.Apoptotic cells were detected by Tunel assays and AnnexinⅤ-FITC apoptosis detection kit.The intracellular ATP level, the change of mitochondrial membrane potential and the mitochondrial DNA oxidative damage were also determined in these groups. Results: (1) The apoptosis rate of H/R group(26.36±5.14)% was significantly higher compared to control group(2.71±1.66)%(P=0.000 4), which could be significantly reduced in S+ H/R group(17.28±4.75)%(P=0.012 8 vs. H/R group , P=0.003 9 vs. control group). The ratio of AnnexinⅤ and PI double positive cells in H/R group(28.23±6.73)% was significantly higher compared to control group(3.53±2.83)%(P=0.001 1), which was significantly reduced in S+ H/R group(18.10±4.56)%(P=0.037 2 vs. H/R group, P=0.038 3 vs. control group). (2)The ATP level of H9c2 cells in H/R group(49.05±10.12)% was significantly lower than in control group 100%(P=0.000 5), which was significantly increased in S+ H/R group(68.67±13.32)%(P=0.019 9 vs. H/R group). Confocal microscope showed that red fluorescence was dominant in the control group, red fluorescence was significantly reduced, while green fluorescence was significantly increased in H9c2 cells of H/R group and the fluorescence ratio of red to green in H/R group((37.13±8.47)%) was significantly decreased compared to control group (100%, P=0.000 1), fluorescence ratio of red to green was significantly increased in S+ H/R group((63.77±12.32)% vs. H/R group, P=0.007 3). (3)The mitochondrial DNA oxidative damage in different groups: there was only few 8-hydroxyguanine (8-OHdG) expression, which marked as green, in control group, and 8-OHdG expression was significantly upregulated in H/R group, moreover, the 8-OHdG was co-localized with mitochondria.The expression of 8-OHdG was significantly lower in S+ H/R group compared to H/R group. Conclusion Salvianolate can reduce mitochondrial DNA oxidative damage, and protect mitochondrial function, thus inhibit myocardial cell apoptosis and eventually reduce the myocardial H/R-injury in H9c2 cells.