Objective To investigate the effect of Hedyotis diffusa extract(HDE)on the proliferation of liver cancer cells and its relationship with sugar metabolism reprogramming and oxidative phosphorylation and analyze its possible mechanisms.Methods CCK-8 and EDU experiments were used to determine the effect of different concentrations(20,40,80 mg/mL)of HDE on the growth of liver cancer cell line SNU-368.Lactate dehydrogenase activity,glucose uptake,lactate production,extracellular pH,mitochondrial respiratory chain complex activity,and cellular oxygen consumption were measured to analyze the effect of HDE on aerobic glycolysis and oxidative phosphorylation in liver cancer cells.qRT-PCR experiments were used to detect the mRNA expressions of GLUT1,GLUT4,HK2,GPI,PFKL,ALDOA and HIF-1α in SNU-368 cells of different groups.Western blotting experiments were used to detect the protein expression of HIF-1α.A stable cell line overexpressing HIF-1αwas constructed by lentivirus transfection of liver cancer cells SNU-368 and then intervened with HDE;the expression of HIF-1α mRNA and protein was detected with qRT-PCR and Western blotting.Results CCK-8 results showed that the HDE exhibited a concentration-dependent inhibitory effect on the proliferation of liver cancer cells(all P＜0.05).Results from glucose metabolism-related tests indicated that the HDE could inhibit glucose uptake and lactate production,decrease lactate dehydrogenase activity,increase extracellular pH value,enhance cellular oxygen consumption,and elevate activities of mitochondrial respiratory chain complexes Ⅰ,Ⅱ,Ⅲ and Ⅳ(all P＜0.05).qRT-PCR results revealed that the HDE suppressed the mRNA expressions of GLUT1,HK2,GPI,and ALDOA(all P＜0.05).qRT-PCR and Western blotting experiments showed that compared to the control group,the expression of HIF-1α mRNA and protein in the HDE group was significantly reduced.However,when HIF-1α was overexpressed and HDE was added in the HIF-1α-LV group,the expression of HIF-1α mRNA and protein increased again compared to the HDE group.Conclusion HDE inhibits glycolysis and promotes oxidative phosphorylation to inhibit the proliferation of liver cancer cells,and its mechanism of action may be related to the inhibition of HIF-1α expression.

Objective To investigate the effect and mechanism of Twist-related protein 1(TWIST1)on pulmonary vascular remodeling induced by monocrotaline(MCT)in pulmonary arterial hypertension(PAH)rats.Methods A total of 50 healthy male Sprague Dawley(SD)rats were randomly divided into five groups including control group,MCT-treated group,MCT and dimethyl sulfoxide(DMSO)-treated group,MCT and harmine-treated group MCT and hydroxychloroquine(HCQ)-treated group.The right ventricle systolic pressure(RVSP)was measured,right ventricular hypertrophy index(RVHI)and percentage of medial wall thickness(MT％)to assess the development of PAH.The protein levels of TW1ST1,autophagy markers LC3B and RND3 were determined using western blot.Results Compared with control group,expressions of TWIST1 and LC3B were increased by 2.32±0.22 folds and 0.87±0.19 folds in MCT-induced PAH group,with significant differences(t=15.812,11.227,all P＜0.00 1),while the protein level of RND3 in MCT-induced PAH rats was decreased by 0.32±0.07 folds compared with control group,with significant difference(t=-13.003,P＜0.001).Administration of TWIST1 inhibitor Harmine or autophagy inhibitor hydroxychloroquine significantly suppressed MCT-induced increase in LC3B and down-regulation of RND3 expression,and reduced RVSP,RVHI and MT％expressions in MCT-induced PAH rats,with significant differences(t=-24.277～16.636,all P＜0.001).Conclusion TWIST1 promotes pulmonary vascular remodeling by inducing autophagy activation,thus promoting the occurrence and development of PAH.

Objective:To study the efficacy and safety of anlotinib in the treatment of advanced radioactive iodine-refractory differentiated thyroid cancer (RAIR-DTC) and the effect of anlotinib on iodine uptake of lesions.Methods:A retrospective analysis was performed on 23 patients (10 males and 13 females, age (59.1±8.7) years) with advanced RAIR-DTC who were treated with anlotinib in Liaocheng Hospital Affiliated to Shandong First Medical University between January 2019 and February 2023 and met the enrollment criteria. Thyroid function, serum thyroglobulin (Tg) and Tg antibody (TgAb) were determined every 6 weeks during the treatment with anlotinib, and maximum diameter of target lesion (TL) was monitored by CT every 12 weeks, in order to evaluate the therapeutic efficacy, and treatment-related adverse reactions were observed. Diagnostic 131I whole body scan (Dx-WBS) was performed in some patients to evaluate the changes in iodine uptake of lesions after anlotinib treatment. In this study, the posttreatment changes of patients within 24 weeks during the treatment were analyzed. The maximum diameter of TL and Tg at different time points were compared by Friedman test, and were further compared in pairs with P values corrected by Bonferroni method. Results:After 24 weeks of treatment with anlotinib, 8 of 23 patients achieved partial response, 15 had stable disease, and no patients achieved complete response. Serum Tg levels at 6, 12, 18, 24 weeks after anlotinib treatment were 189.5(85.0, 483.3), 127.7(52.4, 319.8), 82.0(40.2, 213.5) and 80.1(39.9, 205.0) μg/L, all of which were lower than the baseline level of Tg (384.5(210.9, 1 605.0) μg/L; χ2 values: 4.23-7.86, all P<0.001). Tg level at 18 weeks after treatment was statistically different from that at 12 weeks after treatment ( χ2 =3.06, P<0.001), but was not statistically different from that at 24 weeks after treatment ( χ2 =12.57, P=0.059). The maximum TL diameters of lung and cervical lymph nodes were significantly reduced at week 12 and 24 of anlotinib treatment compared with baseline ( χ2 values: 14.76-31.12, all P<0.001), while there was no significant difference in TL maximum diameter at 12 and 24 weeks of treatment ( χ2 values: 5.65, 9.02, P values: 0.314, 0.070). Common adverse reactions included hypertension, hand-foot syndrome, hyperacylglyceremia and proteinuria. No adverse reactions above grade 4 or death related to adverse reactions occurred. Dx-WBS evaluation was performed in 7 patients after anlotinib treatment, and no change in iodine uptake was found. Conclusions:Anlotinib has a clear effect on advanced RAIR-DTC with less adverse reactions. The efficacy of anlotinib reaches the strongest at around 12-18 weeks and becomes stable at 24 weeks. No effect of anlotinib on inducing redifferentiation of RAIR-DTC cells and enhancing iodine uptake is found.

Objective:To describe the clinicopathological features of pulmonary artery intimal sarcoma (PAIS), and to understand its molecular alterations.Methods:Sixty cases of pulmonary artery endarterectomy performed at the China-Japan Friendship Hospital, Beijing, China from January 2017 to January 2020 were reviewed. Clinical data of 5 patients with pulmonary artery intimal sarcoma were collected. Hematoxylin-eosin staining, immunohistochemistry staining and fluorescence in situ hybridization (FISH) were performed to evaluate the pathological features. RNA sequencing was conducted to assess the fusion gene changes in PAIS.Results:The detection rate of PAIS was 8.3% (5/60), with the median age of 49 years and a female predominance. Their clinical manifestations were non-specific. Histopathological examination showed that the tumors were composed of malignant spindle or epithelioid cells, with various degrees of atypia. Focal heterologous osteosarcomatous or leiomyosarcomatous differentiation was noted. The tumor cells could express PDGFRA, CDK4 and MDM2 with co-amplification of MDM2, CDK4 and EGFR genes. RNA sequencing detected multiple in-frame fusions in the tumors.Conclusions:PAIS is a rare, highly heterogeneous, and poorly-or un-differentiated sarcoma accompanied by complex changes of multiple genes.It has no known effective treatments, and thus has a poor prognosis.

Objective: Understanding mental health status of students with learning disabilities in Beijing to provide a basis for mental health promotion of students with learning disabilities. Methods: By means of random cluster sampling, 5 787 enrolled students in grade one and grade two of 11 public junior middle schools in Beijing were selected as the survey subjects. A self designed questionnaire was used to investigate the students’ learning disabilities and mental health status through anonymous self filling. Results: About 11.6% students self reported learning disabilities. The proportions of students with learning difficulty in mathematical reasoning and calculation were higher, 44.1% and 40.7% respectively. The reported rate of mental health problems was 38.3%. The top four of the 10 symptoms were obsessive compulsive symptoms, learning pressure, emotional instability and anxiety(2.19±0.77)(2.17±0.99)(2.09±0.90)(2.07±1.08). Compared with students without learning disabilities, students with self reported learning disabilities had poorer mental health status(OR=1.47), and learning disabilities were related to most factors of mental health. Different types of learning disabilities were associated with different mental health factors. Conclusion Mental health problems of students with learning disabilities are higher than that of students without learning disabilities. It is necessary to strengthen the mental health support for students with learning disabilities and improve their mental health.

Objective: To investigate the relationship between V-raf murine sarcoma viral oncogene homologue B1 (BRAF)^V600E mutation and metastasis after thyroidectomy in papillary thyroid carcinoma (PTC) with different recurrence risk stratification. Methods: From March 2014 to September 2017, 134 PTC patients (45 males, 89 females; age: 16-72 years) who underwent ¹³¹I treatment in the Department of Nuclear Medicine of Liaocheng People′s Hospital and had undergone BRAF^V600E mutation detection were retrospectively analyzed. The recurrence risk during surgery was divided into 3 levels: low-, medium- and high-risk. Each recurrence risk group was divided into 3 subgroups according to the postoperative follow-up results: non-metastasis group, cervical lymph node metastasis group and pulmonary metastasis group. BRAF^V600E mutation rates in different groups were compared (χ² test). Results: The BRAF^V600E mutation rate was 55.22%(74/134) in 134 PTC patients. The mutation rates were not significantly different in the 3 metastasis subgroups for low-risk patients (n=46; χ²=2.39, P>0.05). In medium-risk patients (n=47), the mutation rate in neck lymph node metastasis group (16/19) was higher than that in pulmonary metastasis group (3/8) and non-metastasis group (25.00%, 5/20; χ² values: 5.89 and 13.75, both P<0.05), while there was no difference between pulmonary metastasis group and non-metastasis group (χ²=0.44, P>0.05). In high risk patients (n=41), the mutation rate in neck lymph node metastasis group (85.00%, 17/20) was higher than that in the pulmonary metastasis group (5/13) and non-metastasis group (1/8; χ² values: 7.68 and 13.08, both P<0.01), while there was no difference between pulmonary metastasis group and non-metastasis group (χ²=1.64, P>0.05). Conclusion The BRAF^V600E mutation is closely related to neck lymph node metastasis after operation in middle- and high-risk patients with PTC, but it does not significantly increase the probabilities of neck lymph node metastasis in low-risk patients and pulmonary metastasis in low-, medium- and high-risk patients.

Objective To study and the succession of sarcosaphagous insects and their regular activity on carcass in Xinxiang area. Methods Six rats and five rabbits were sacrificed and placed at the remote grassland of Xinxiang Medical University in July 2015. The main insect species and the decomposition process were observed. The different decomposition stages and the community succession of insects were recorded. Results Seven main species could be identified belonging to 3 families from Diptera, main including Musca domestica, Chrysomya megacephala, Lucilia sericata, Boettcherisca peregrina, Helicophagella melanura. Muscina stabulans and Fannia prisca. And showed regulations of the succession of sarcosaphagous flies community. Conclusion It may be used for estimating postmortem interval in Xinxiang area.

Objective:To summarize the major risk factors of infection in the process of cleaning and disinfection of endoscopy center in our hospital,and formulate the corresponding prevention measures to reduce the incidence of endoscopic infection so as to ensure medical safety of patients.Methods:The specialist check the cleaning disinfection process,statistics of each link in the risk factors of infection.600 endoscopes were taken at random after disinfection,endoscopic inner cavity and the outer surface samples are collected,the cleaning and disinfection of the test results for evaluating quality indicators.Results:Testing 600 samples of qualified 584,and the percent of pass is 97.3％,gastroscopy,colonoscopy,duodenum mirror,bronchoscope,endoscopic ultrasonography qualified rate was 98.6％,96.8％,95.6％,96.0％,97.2％,respectively.28 strains of pathogenic bacteria were detected in 16 cases of unqualified samples,including 13 strains of helicobacter pylori,7 strains of escherichia coli,4 strains of pseudomonas aeruginosa,2 strains of pseudomonas aeruginosa,1 strains of staphylococcus aureus,1 strains of Klebsiella pneumoniae;16 cases to detect unqualified reason analysis,scrub incomplete accounted for 37.5％,Not according to the instructions required to use multi-enzyme and disinfectant accounted for 18.75％,special infected patients did not do special treatment of endoscopy accounted for 12.5％,endoscopic insufficient drying and the staffhand hygiene is not up to standard accounted for 12.5％,respectively.Conclusion:Endoscopic cleaning and disinfection aspects of infection caused by many factors,infection control education should be strengthened for all personal in the endoscopy center,regular professional training of cleaning and sanitation personal,cleaning and disinfection process should be standardized management,establish traceability registration system,control the quality of the endoscope cleaning and disinfection from each link.It is the main measure to prevent digestive endoscopy center in infection.

Objective:To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein.Methods:A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured;the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry.Results:Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P<0.05 or P<0.01);the indexes metioned above of the mice in MUC1-MBP+ R848 were higher than those in MUC1-MBP+BCG group;the IL-4 levels had no significant difference(P>0.05).Compared with control group,the proprotions of CD3+T,CD4+T,and CD8+ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01).Conclusion:Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissues on MFC gastric cancer bearing mice; To discuss relevant mechanism of action. Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study. A total of eligible 48 mice were randomly divided into model group, DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups, modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. The treatment was conducted once a day, and lasted for 14 continuous days. After the last administration of gavage orally treatment, all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissue with the method of RT-qPCR and immunohistochemistry. Meanwhile, the tumor growth was observed and the general conditions of mice were recorded. Results The model group was rich in tumor cells; the sizes of cells were different; the volume was large; the nucleus was deeply stained and the heterotypic shape was obvious, and the small focal necrosis was seen. The number of tumor cells in each administration group was reduced; the arrangement was loose; the cell volume was significantly reduced, and the nuclear pyknosis was reduced. Cell necrosis significantly increased; the number of interstitial blood vessels decreased; collagen fibers increased, especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. Compared with the model group, the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 mRNA and protein decreased in each administration group. TLR2, TLR4, TLR6, TRAF6 and MyD88 were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups, the protein changes were more obvious Compared with DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups. Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2, TLR4, TLR6, TRAF6 and MyD88 expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.