ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

Objective:To study the clustered regularly interspaced short palindromic repeats (CRISPR) genotype of Yersinia pestis and its regional distribution characteristics in natural plague focus of Tibet Autonomous Region. Methods:A total of 125 representative Yersinia pestis strains isolated from natural plague focus in Tibet Autonomous Region at different times, regions, hosts and vectors were selected as experimental strains, and the phenol chloroform mixed extraction method was used to extract Yersinia pestis DNA. Three pairs of CRISPR primers (for YPa, YPb, YPc locus) were used to amplify the DNA of the experimental strains, and the CRISPR genotype of Yersinia pestis was determined by sequencing. Results:All 125 strains of Yersinia pestis had three CRISPR locus: YPa, YPb, and YPc. A total of 18 spacer were found, including 8 in YPa loci, 6 in YPb loci, and 4 in YPc loci. Two new types of spacers had been discovered, namely b52 and c14. CRISPR typing revealed 10 genotypes, including G1, G7, G7-b4''', G7-b52, G7-c2 -, G8, G22, G22-a4 -, G22-b4''', and G22-c14, of which 6 were newly discovered genotypes. Among the 125 experimental strains, G7 was the main genotype, accounting for 65.6% (82/125), which was distributed in 6 prefecture level citys and 1 region of Tibet Autonomous Region. Next were G22 and G7-c2 - genetypes, accounting for 14.4% (18/125) and 11.2% (14/125), respectively. G22 gene type was distributed in Nagqu, Changdu, Lhasa citys, and Ngari Prefecture, while G7-c2 - genetype was distributed in Shigatse and Shannan cities. Conclusion:The CRISPR locus of Yersinia pestis in natural plague focus of Tibet Autonomous Region is highly polymorphic, and the Yersinia pestis strains with different genotypes have obvious regional distribution characteristics.

ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.

Objective:To investigate the effects of biologics on psychological status and quality of life in patients with inflammatory bowel disease (IBD).Methods:A cross-sectional survey was conducted in 42 hospitals in 22 provinces (autonomous regions and municipalities directly under the central government) from September 2021 to May 2022. General clinical information and the use of biologics were obtained from adult patients diagnosed with IBD who voluntarily participated in the study. Psychological status was evaluated using the Generalized Anxiety Disorder (GAD-7), Patient Health Questionnaire-9 (PHQ-9), Pittsburgh Sleep Quality Index (PSQI), and Inflammatory Bowel Disease Questionnaire (IBDQ) assessment tools. Counts were analyzed via the Chi-square test, and datasets that were not normally distributed were analyzed via nonparametric tests. P<0.05 was considered statistically significant. Results:A total of 2 478 valid questionnaires were collected. The GAD-7 score of the biologics group was significantly lower than that of the non-use group [6 (2, 9) vs. 7 (3, 10), Z=-3.49, P<0.001]. IBDQ scores [183 (158, 204) vs. 178 (152, 198), Z=-4.11, P<0.001], intestinal symptom scores [61 (52, 67) vs. 58 (49, 65), Z=-5.41, P<0.001], systemic symptom scores [28 (24, 32) vs. 27 (23, 31), Z=-2.37, P=0.018], emotional ability scores [69 (58, 77) vs. 67 (56, 75), Z=-3.58, P<0.001] and social ability scores [26 (22, 29) vs. 25 (22, 29), Z=-2.52, P=0.012] in the biologics group were significantly higher than in the non-use group. GAD-7 scores [5 (2, 9) vs. 6 (3, 10), Z=-3.50, P<0.001] and PSQI scores [6 (4, 9) vs. 6 (4, 9), Z=-2.55, P=0.011] were significantly lower in the group using infliximab than in the group not using it. IBDQ scores were significantly higher in patients using vedolizumab than in those not using it [186 (159, 205) vs. 181 (155, 201), Z=-2.32, P=0.021] and were also significantly higher in the group treated with adalimumab than in the group not treated with adalimumab [187 (159, 209) vs. 181 (155, 201), Z=-2.16, P=0.030]. However, ustekinumab had no significant effect on any of the scores. Conclusion:The use of biologics is strongly associated with improvements in anxiety status and quality of life in IBD patients.

Objective:To investigate whether miR-548b-3p can regulate the malignant biological traits of hepatocellular carcinoma cells by down-regulating the expression of decoy receptor 3( DcR3). Methods:The miR-548b-3p levels in 5 common hepatoma cell lines Hep3b, HepG2, Huh7.5.1, PLC and 97H were detected by real-time fluorescence quantitative PCR. The experimental hepatoma cell lines were purchased from Shanghai Cancer Institute. The subcutaneous tumor formation experiment in nude mice included two parts: miR-548b-3p over expression group and down expression group. miR-548b-3p over expression experiment included Huh7.5.1- miR-548b-3p cell group and Huh7.5.1-NC cell group. The experimental group with low expression of miR-548b-3p included PLC- miR-548b-3p-sponge group and PLC-NC cell group, with 5 nude mice in each group. The nude mice used in the experiment were male Balb/c strain, body weight 20-25 g, 4-6 weeks old. Starbase software was used to predict whether there were binding sites between miR-548b-3p and DcR3. Dual luciferase reporting assay to verify whether DcR3 is the target gene of miR-548b-3p. Rescue experiments were conducted to verify whether the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Statistical analysis was performed using Graphpad Prism 9. Measurement data with normal distribution were expressed as mean±standard deviation( ± s), and t-test was adopted for comparison between general measurement data groups. Results:The relative expression level of 2 -ΔΔCt was used as the parameter for real-time fluorescence quantitative PCR comparison. Hep3b expression level was 1, HepG2 expression level was 0.902, Huh7.5.1 expression level was 0.712, PLC expression level was 1.293, and 97H expression level was 0.818. The final results showed that, The highest expression of miR-548b-3p was in PLC cells, and the lowest expression was in Huh7.5.1 cells. The subcutaneous tumor formation experiment of nude mice showed that after 4 weeks, the tumor volume of Huh7.5.1- miR-548b-3p was(444.77±142.34) mm 3, and that of Huh7.5.1-NC was(918.80±139.21) mm 3. The tumor volume of PLC- miR-548b-3p-sponge was(407.49±58.50) mm 3, and that of PLC-NC was(218.62±47.55) mm 3. The binding sites of miR-548b-3p and DcR3 were predicted by Starbase software. Dual luciferase assay showed that DcR3 was the target gene of miR-548b-3p. Rescue experiments verified that the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Conclusion:miR-548b-3p can regulate the proliferation, invasion, migration and other malignant biological traits of liver cancer cells, and it is achieved by down-regulating the expression level of DcR3.

Objective To investigate the safety,feasibility,and efficacy of highly selective protective vagotomy in laparoscopic fundoplication.Methods Clinical data of 78 patients who underwent laparoscopic hiatal hernia repair plus fundoplication(short floppy Nissen procedure)for gastroesophageal reflux disease and hiatus hernia from January 2014 to December 2019 in our hospital were retrospectively analyzed.The patients were divided into two groups:the traditional operation group and the vagus nerve protection group.The operation time,blood loss during operation,hospital stay after operation and the incidence of postoperative complications were compared between the two groups.The GERD Q score,DeMeester score,lower esophageal sphincter pressure(LESP),and control of reflux symptoms at 6 months after operation in the two groups were analyzed.Results Both groups of surgeries were successfully completed,and there were no serious intraoperative side injuries.There was no significant difference between the traditionaloperationgroupandthevagusnerveprotectiongroupinoperationtime[(85.5±13.9)minvs.(88.3±18.6)min,t =0.729,P =0.468],intraoperative blood loss[(18.6±8.6)ml vs.(18.1±8.5)ml,t =-0.221,P =0.825],and postoperative transanal exhaust time[(2.0±0.7)d vs.(1.8±1.0)d,t =-1.227,P =0.224].The postoperative hospital stay in the traditional surgical group was significantly longer than that in the vagus nerve protection group[(9.4±3.0)d vs.(8.2±2.1)d,t =-2.172,P = 0.033].The incidence of surgical complications within 30 d after surgery in the traditional surgical group was 36.8%(14/38),which was significantly higher than that in the vagus nerve protection group[12.5%(5/40),χ2 = 6.267,P = 0.012].The traditional surgical group had a cure rate of 86.8%(33/48)at 6 months after surgery,which was not significantly different from the vagus nerve protection group[85.0%(34/40),Z =-0.232,P =0.816].There were no significant differences in GERDQscore,DeMeester score,LESP between the two groups at 6 months after surgery[(5.6±0.9)points vs.(5.8±0.8)points,t =1.232,P =0.222;(4.1±2.2)points vs.(4.2±2.2)points,t =0.261,P =0.795;(23.2±3.5)mm Hg vs.(23.5±3.8)mm Hg,t = 0.412,P = 0.681].Conclusion It is safe,feasible,and effective to apply the highly selective protective vagotomy in laparoscopic short floppy Nissen fundoplication to protect the vagus nerve.

Objective:To investigate the expression of anti apoptosis inhibitor antibody in serum of patients with advanced primary liver cancer and its relationship with the prognosis after transcatheter hepatic arterial chemoembolization (TACE).Methods:One hundred and three patients with advanced primary liver cancer were selected from our hospital and treated with TACE. Serum anti-survivin antibody expression levels were detected 1 day before surgery and 1 month after surgery. To analyze the relationship between serum anti-survivin antibody level and short-term therapeutic effect, clinicopathological features and prognosis were analyzed.Results:the level of anti-survivin antibody in patients with disease remission was significantly lower than that in patients without disease remission (81.84±9.30 vs. 90.84±10.21, P<0.05), and the change of anti-survivin antibody in patients with disease remission was significantly higher than that in patients without disease remission (30.93±5.63 vs. 22.75±4.52, P<0.05). The changes of anti-survivin antibody before and after TACE were correlated with TNM stage, maximum tumor diameter and degree of differentiation ( P<0.05). The results of survival analysis showed that the postoperative survival of patients with △ reduced anti-survivin antibody was significantly better than that of patients without △ reduced anti-survivin antibody ( P<0.05). The area under the ROC curve was 0.850 in the prediction of one-year death value of patients with primary liver cancer by △ anti-survivin antibody. Conclusion:the difference of anti survivin antibody before and after TACE in patients with advanced liver cancer is closely related to the short-term and long-term prognosis.

ObjectiveTo investigate the late detection of HIV/AIDS cases in the elderly in Jilin Province and analyze its influencing factors， to provide theoretical basis for improving their life quality. MethodsThe first CD4 values of HIV/AIDS patients aged 50 years and above living in Jilin Province were used to estimate late detection， and the influencing factors of late detection in elderly cases were analyzed. ResultsThe average CD4 cell count of newly reported HIV/AIDS cases aged 50 and above in Jilin Province from 1996 to 2021 was （230.55±191.97）， the low value group accounted for the largest proportion （50.8%）， and the late detection rate was 59.3% （1397/2325）. The late detection cases were mainly from sexual transmission （46.8% for same-sex and 48.2% for heterosexual contact）. From the perspective of sample sources， most of the late detection patients were diagnosed while testing for other illnesses， followed by testing and consulting. In terms of contact history， the late detection of cases of men who have sex with men was higher. The binary logistic regression analysis showed that gender， marriage， sample source and report year were the factors affecting the late detection of AIDS. The late detection rate of males was higher， and cases of married couples were more likely to be late detection. With the increase of report year， the late detection rate decreased， and testing and counseling could effectively reduce the late detection rate of AIDS. ConclusionThe CD4 cell count in the first detection of HIV/AIDS in the elderly in Jilin Province is low， and the late detection rate of male cases is high. In recent years， the expansion of voluntary counseling and testing in Jilin Province has effectively reduced the late detection rate of HIV/AIDS. At the same time， sex education should be strengthened for the elderly， healthy marital relationships should be advocated and more attention should be paid to the mental health of the elderly.

Objective:To propose a markless patient setup workflow based on the optical surface monitoring system (AlignRT) and open-face mask immobilization for whole-course head tumor radiotherapy, assess the setup time and repositioning frequency of the proposed workflow, and conduct a comparative analysis of the differences, correlation, and consistency of the setup errors of the AlignRT and cone beam CT (CBCT) systems.Methods:A retrospective analysis was conducted for the data on the errors of 132 fractionated setup based on open-face mask immobilization of 33 head tumor patients. AlignRT-guided markless patient setup workflow was applied throughout the radiotherapy. Meanwhile, the body structures automatically generated by the treatment planning system were used as body references. The 6-degree-of-freedom (6DoF) setup errors (lateral, vertical, longitudinal, rotation, pitch, roll, and yaw directions), setup time, and repositioning frequency of the AlignRT and CBCT systems were recorded and analyzed. The Wilcoxon and Spearman analyses were used to statistically assess the differences and correlation of the setup errors of the two systems. Moreover, the Bland-Altman analysis was employed to evaluate the consistency of the two systems.Results:The 6DoF setup errors of CBCT were within the clinical tolerance (linear motions: -0.30 to 0.30 cm; rotational motions: -2.0° to 2.0°). The setup time and repositioning frequency of CBCT were (98 ± 31) s and 1.51% (2/132), respectively. There was no significant difference in setup errors between the two systems except those in x-axis ( Z = -3.11, P= 0.002), y-axis ( Z = -7.40, P<0.001), and Pitch ( Z= -4.48, P<0.001). There was a significant positive correlation between the setup errors along lateral ( rs = 0.47, P<0.001) and vertical ( rs = 0.29, P = 0.001) directions, rotation (Rtn; rs = 0.47, P<0.001), pitch (Pitch; rs = 0.28, P = 0.001) and roll (Roll; rs = 0.45, P<0.001) of the two systems. The 95% limits of agreement (95% LoA) of 6DoF setup errors were -0.12 to 0.09 cm, -0.07 to 0.17 cm, -0.19 to 0.20 cm, -1.0° to 0.9 °, -1.0° to 1.5°, and -0.9° to 1.0°, respectively. The 95% confidence interval (95% CI) of 95% LoA was -0.14 to 0.11 cm, -0.09 to 0.19 cm, -0.23 to 0.23 cm, -1.2° to 1.1°, -1.2° to 1.7°, and-1.0° to 1.1°, respectively, all of which were within the permissible error ranges. The 6DoF setup error difference of 3.41% (27/792< 5%) was beyond the 95% LoA. The maximum absolute differences of 6DoF setup errors within the 95% LoA were 0.12, 0.16, 0.19 cm, 0.9°, 1.5°, and 1.0°, respectively. Conclusions:The proposed markless setup workflow based on AlignRT combined with open-face mask immobilization for whole-course head tumor radiotherapy exhibits reasonable agreement and consistency with the patient setup using CBCT, with acceptable clinical efficiency. It can be applied to the first radiotherapy and the real-time monitoring of therapy to improve the safety and thus is of value in clinical applications.