ObjectiveTo investigate the mechanism of Forsythiae Fructus-Lonicerae Japonicae Flos（FF） in the treatment of acute lung injury（ALI） by investigating the effects of FF on serum metabolomics of rats with ALI. MethodsThirty male SD rats were acclimated for 1 week， and 6 rats were randomly selected as the blank group. The other 24 rats were injected with lipopolysaccharide（LPS） solution by tracheal drip to establish an ALI model. After successful model establishment， the rats were randomly divided into the model group， the FF low-dose group（3.0 g·kg^-1）， the FF high-dose group（6.0 g·kg^-1）， and the dexamethasone group（5 mg·kg^-1）， with six rats in each group. The FF low- and high-dose groups and the dexamethasone group were received daily oral administration of the corresponding drug solution， and the blank group and the model group were gavaged with an equal amount of saline， treatment was administered continuously for 3 d. The pathological conditions of rat lung tissues were evaluated by hematoxylin-eosin（HE） staining， wet/dry mass ratio（W/D） of the lung tissues， and protein concentration in rat bronchoalveolar lavage fluid（BALF）. Metabolomic analysis of rat serum was performed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， combined with multivariate statistical analysis， the potential biomarkers of FF in treating ALI were screened by variable importance in the projection（VIP） value>1， P<0.05 from t-test， and log₂fold change（FC）>1 or log₂FC<-1. Kyoto Encyclopedia of Genes and Genomes（KEGG） database combined with MetaboAnalyst were used for pathway analysis of the screened differential metabolites. The protein expression levels of sphingosine-1-phosphate（S1P）， phosphatidylinositol 3-kinase（PI3K）， protein kinase B1（Akt1）， and phosphorylated Akt1（p-Akt1） were examined by Western bolt. The expression levels of interleukin（IL）-6， IL-1β， and tumor necrosis factor（TNF）-α in BALF were detected by enzyme-linked immunosorbent assay（ELISA）. ResultsCompared with the blank group， rats in the model group showed ALI pathological features such as alveolar lumen dilatation， interstitial hemorrhage and massive inflammatory cell infiltration， and the protein concentration in BALF and W/D of the lung tissues were significantly elevated（P<0.01）. Compared with the model group， the low- and high-dose groups of FF as well as the dexamethasone group exhibited reduced pulmonary bronchial hemorrhage in rats， and the protein concentration in BALF and W/D were significantly decreased（P<0.05）， and the lung injury was significantly alleviated. Analysis of rat serum metabolomics revealed that FF downregulated 38 biomarkers. Pathway enrichment analysis showed that FF primarily exerted therapeutic effects through 7 key metabolic pathways， including arginine biosynthesis， sphingomyelin metabolism， alanine， aspartate and glutamate metabolism， taurine and hypotaurine metabolism， α-linolenic acid metabolism， niacin and nicotinamide metabolism， and retinol metabolism. The results of Western bolt and ELISA showed that， compared with the blank group， the model group exhibited significantly elevated expression levels of S1P， PI3K， Akt1 and p-Akt1 proteins in the lung tissues， as well as increased expression levels of IL-6， IL-1β and TNF-α in BALF（P<0.01）. Compared with the model group， the expression levels of the aforementioned indicators were significantly downregulated in the low- and high-dose FF groups as well as the dexamethasone group（P<0.05， P<0.01）. ConclusionFF may play a role in ALI by regulating amino acid metabolism and lipid metabolism， and its mechanism may be related to the inhibition of S1P/PI3K/Akt1 signaling pathway to attenuate the inflammatory response caused by ALI.

ObjectiveSerum metabolomics of acute lung injury（ALI） in rats was conducted using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） to explore the similarities and differences in the mechanism of Qingqiao（harvested when the fruits of Forsythiae Fructus were initially ripe and still green in color） and Laoqiao（harvested when the fruits of Forsythiae Fructus were ripe） in the treatment of ALI. MethodA total of 24 SD male rats were acclimatized and fed for 1 week， 6 of them were randomly selected for the blank group and 18 for the experimental group. The ALI model was induced in the experimental group by tracheal intubation with lipopolysaccharide（LPS）. After successfully constructing the ALI model， these rats was randomly divided into model group， Qingqiao group and Laoqiao group， with 6 rats in each group. The Qingqiao and Laoqiao groups were administered orally once a day at a dose of 1.5 g·kg^-1， while the blank and model groups received an equivalent volume of saline for 3 consecutive days. The pathological conditions of rat lung tissues were comprehensively assessed by hematoxylin-eosin（HE） staining， wet-to-dry mass ratio（W/D） of lung tissues， and protein concentration in rat bronchoalveolar lavage fluid（BALF）. The levels of interleukin（IL）-6， IL-1β and tumor necrosis factor（TNF）-α in BALF were quantified using enzyme-linked immunosorbent assay（ELISA）. UPLC-Q-TOF-MS was used to identify and analyze the chemical compositions of Qingqiao and Laoqiao， and serum metabolomics of rats in each group was analyzed， combined with multivariate statistical analysis with variable importance in the projection（VIP） value>1， P<0.05 from t-test， and fold change（FC）≥1.5 or FC≤0.5 to screen the differential metabolites Qingqiao and Laoqiao for the treatment of ALI. The Kyoto Encyclopedia of Genes and Genomes（KEGG） database was used in combination with MetaboAnalyst for the metabolic pathway analysis of the screened differential metabolites. ResultCompared with the blank group， rats in the model group exhibited enlarged alveolar lumen， ruptured alveoli， interstitial hemorrhage， bronchial exudation of a large number of neutrophils and erythrocytes， and a significant increase in the protein concentration in the BALF and the W/D value of the lung tissues（P<0.01）. In contrast， compared with the model group， rats in the Qingqiao group and the Laoqiao group showed reduced bronchial hemorrhage in the lungs， and the protein concentration in the BALF and the W/D value of the lung tissues were significantly decreased（P<0.01）， the lung injury was significantly alleviated， but more obvious in the Qingqiao group. Compared with the blank group， the expression levels of IL-6， IL-1β and TNF-α in the BALF of the model group were significantly higher（P<0.01）. Additionally， compared with the model group， the expression levels of IL-6， IL-1β and TNF-α in the Qingqiao and Laoqiao groups were significantly lower（P<0.01）. The chemical composition analysis of Qingqiao and Laoqiao revealed that 63 components were detected in Qingqiao and 55 components were detected in Laoqiao， with 47 common components， 16 components unique to Qingqiao and 8 components unique to Laoqiao. Characterizing the differences in serum metabolomics in rats， 19 and 12 metabolites were called back by Qingqiao and Laoqiao， respectively. The metabolic pathway enrichment analysis showed that Qingqiao exerted its therapeutic effects by affecting 6 key metabolic pathways， including linoleic acid metabolism， phenylalanine metabolism， phenylalanine， tyrosine and tryptophan biosynthesis， glycerophospholipid metabolism， α-linolenic acid metabolism， and arachidonic acid metabolism， and Laoqiao exerted therapeutic effects by affecting 6 key metabolic pathways， including linoleic acid metabolism， arachidonic acid metabolism， sphingolipid metabolism， phenylalanine metabolism， ascorbate and aldarate metabolism， and glycerophospholipid metabolism. ConclusionQingqiao and Laoqiao have therapeutic effects on ALI， and Qingqiao is more effective. Both of them can play a therapeutic role in ALI by regulating amino acid metabolism and lipid metabolism， but the metabolic pathways affected by them are different.

OX40 is a costimulatory receptor that is expressed primarily on activated CD4⁺, CD8⁺, and regulatory T cells. The ligation of OX40 to its sole ligand OX40L potentiates T cell expansion, differentiation, and activation and also promotes dendritic cells to mature to enhance their cytokine production. Therefore, the use of agonistic anti-OX40 antibodies for cancer immunotherapy has gained great interest. However, most of the agonistic anti-OX40 antibodies in the clinic are OX40L-competitive and show limited efficacy. Here, we discovered that BGB-A445, a non-ligand-competitive agonistic anti-OX40 antibody currently under clinical investigation, induced optimal T cell activation without impairing dendritic cell function. In addition, BGB-A445 dose-dependently and significantly depleted regulatory T cells in vitro and in vivo via antibody-dependent cellular cytotoxicity. In the MC38 syngeneic model established in humanized OX40 knock-in mice, BGB-A445 demonstrated robust and dose-dependent antitumor efficacy, whereas the ligand-competitive anti-OX40 antibody showed antitumor efficacy characterized by a hook effect. Furthermore, BGB-A445 demonstrated a strong combination antitumor effect with an anti-PD-1 antibody. Taken together, our findings show that BGB-A445, which does not block OX40-OX40L interaction in contrast to clinical-stage anti-OX40 antibodies, shows superior immune-stimulating effects and antitumor efficacy and thus warrants further clinical investigation.
Mice ; Animals ; Receptors, Tumor Necrosis Factor/physiology* ; Receptors, OX40 ; Membrane Glycoproteins ; Ligands ; Antibodies, Monoclonal/pharmacology* ; Antineoplastic Agents/pharmacology*

Objective:To compare the knowledge of endoscopic screening of gastrointestinal cancers between digestive healthcare workers and new chatbots (chatGPT and new Bing).Methods:A test with twenty-three questions of endoscopic screening of gastrointestinal cancers was conducted, focusing on the appropriate age of screening, high-risk factors, the follow-up time, and the advantages and risks of digestive endoscopy. Digestive healthcare workers were invited to complete the test through electronic questionnaires. New Bing and chatGPT were used to answer each question for 10 rounds. The primary endpoint was the correct rate of all answers. The answer accuracy between digestive healthcare workers and new chatbots were compared using variance analysis, and the factors that affected the accuracy of the answers in digestive healthcare workers were explored using univariate and multivariable liner regression analysis.Results:The results of the test completed by 76 digestive healthcare workers (21 residents, 28 digestive nurses, and 27 digestive doctors) were analyzed. The accuracies were 36.4%±10.9%, 34.5%±10.2%, 52.2%± 12.6%, 46.3%±9.8% and 67.1%±9.3% in residents, digestive nurses, digestive doctors, chatGPT, and new Bing, respectively, with significant difference ( F=22.6, P<0.001). The accuracy was highest in new Bing ( P<0.001). The accuracy was comparable between chatGPT and digestive doctors (LSD- t=-1.398, P=0.166), and both higher than that of digestive nurses (LSD- t=2.956, P=0.004; LSD- t=5.955, P<0.001) and residents (LSD- t=2.402, P=0.018; LSD- t=4.951, P<0.001). Furthermore, the accuracy was comparable between digestive nurses and residents (LSD- t=-0.574, P=0.567). Compared with new Bing, digestive doctors had lower accuracy in answering questions related to adverse events of screening, follow-up recommendation of intestinal metaplasia, high risk factors and screening methods for colon cancer ( P<0.05), but higher accuracy in answering questions related to endoscopic adverse events and screening methods for esophageal cancer ( P<0.05). Multivariable liner regression analysis showed that being digestive doctors ( β=11.7, t=3.054, P=0.003) and questionnaire response time (≥7.6 min) ( β=7.8, t=2.894, P=0.005) were independent factors for the answer accuracy of digestive healthcare workers. Conclusion:Compared with digestive healthcare workers, New chatbots—new Bing has higher accuracy in answering gastrointestinal cancer screening-related questions, but performs poorly in answering questions such as adverse events of endoscopy and screening methods for esophageal cancer.

Objective:To evaluate the role of succinate dehydrogenase (SDH) in hypoxic postconditioning (HPC)-induced reduction of hypoxia-reoxygenation (H/R) injury in myocardial cells of rats and the relationship with mitochondrial ATP-sensitive potassium channels (mito-K ATP). Methods:Myocardial cells isolated from adult male Sprague-Dawley rats were cultured for 48 h and then divided into 7 groups ( n=24 each) using a random number table method: blank control group (Nor group), H/R group, SDHA-siRNA adenovirus+ H/R group (siRNA+ H/R group), HPC group, SDHA-siRNA adenovirus+ HPC group (siRNA+ HPC group), 5-HD+ HPC group, and SDHA-siRNA adenovirus+ 5-HD+ HPC group (siRNA+ 5-HD+ HPC group). Nor group was continuously cultured for 195 min under normoxic conditions. The H/R injury model was prepared by exposing the cells to hypoxia for 45 min in 5% CO 2 + 1% O 2 + 94% N 2, followed by reoxygenation for 150 min. The HPC method involved three cycles of 5 min reoxygenation/5 min hypoxia at the end of 45 min ischemia before 120 min reoxygenation. The mito-K ATP blocker 5-HD administration method involved adding 5-HD at a final concentration of 100 μmol/L at 30 min of hypoxia. The myocardial cells in each siRNA group were successfully transfected with SDHA-siRNA adenovirus to silence SDHA expression. The cell viability, calcium ion level, SDH activity, ATP content, degree of mitochondrial permeability transition pore (mPTP) opening, and mitochondrial membrane potential (MMP) were measured at the end of reoxygenation. Results:Compared with Nor group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, level of calcium ion and activity of SDH were increased in H/R group ( P<0.05). Compared with H/R group, the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ H/R group and HPC group ( P<0.05). Compared with HPC group, the cell viability, ATP content and MMP were significantly decreased, and the degree of mPTP opening, calcium ion level and SDH activity were increased in 5-HD+ HPC group ( P<0.05), and the cell viability, ATP content and MMP were significantly increased, and the degree of mPTP opening, calcium ion level and SDH activity were decreased in siRNA+ HPC group ( P<0.05). Compared with siRNA+ HPC group, the cell viability, ATP content and MMP were significantly decreased, the opening degree of mPTP and calcium ion level were increased ( P<0.05), and no significant change was found in the SDH activity in siRNA+ 5-HD+ HPC group ( P>0.05). Compared with 5-HD+ HPC group, the SDH activity was significantly decreased, and no significant change was found in the other parameters in siRNA+ 5-HD+ HPC group ( P>0.05). Conclusions:HPC alleviates H/R injury probably by reducing SDH activity and opening mito-K ATP in myocardial cells of rats.

Objective:To study peripheral nerve morphology in patients with transthyretin familial amyloid polyneuropathy (TTR-FAP) using high-frequency ultrasonography (HFUS), and to evaluate the value of HFUS in diagnosis of TTR-FAP.Methods:Thirty-eight patients with TTR-FAP diagnosed by gene detection and 23 normal controls from June 2015 to June 2021 in Peking University First Hospital were enrolled in this study. Consecutive ultrasonography scanning was performed in 6 pairs of nerves of bilateral limbs with 30 sites. The cross sectional area (CSA), CSA variability and inter-nerve CSA variability data of the two groups were retrospectively calculated and compared.Results:Compared with the normal controls, TTR-FAP patients showed larger CSA values at most nerve sites of both upper and lower limbs, and there were statistically significant differences at M1(median nerve) [8.55 (6.90, 9.40) mm 2vs 10.05 (9.10, 14.10) mm 2, Z=5.58, P<0.001], M3 (median nerve) [(6.98±1.66) mm 2vs (9.29±2.30) mm 2, t=6.28, P<0.001], M5 (median nerve) [(8.91±1.81) mm 2vs (14.33±4.20) mm 2, t=9.84, P<0.001], U5 (ulnar nerve) [(6.20±1.93) mm 2vs (9.34±2.85) mm 2, t=7.31, P<0.001], Sci1 (sciatic nerve) [(53.50±17.24) mm 2vs (79.74±20.75) mm 2, t=7.57, P<0.001], Sci2 (sciatic nerve) [(53.66±14.21) mm 2vs (73.98±19.21) mm 2, t=6.82, P<0.001] and Tib (tibial nerve) [(31.05±8.43) mm 2vs (46.29±13.14) mm 2, t=7.84, P<0.001] sites. There was no statistically significant difference in CSA at each site among the different subtypes and disease severity of TTR-FAP patients ( P>0.05). There was no statistically significant difference in CSA-variability of the median and ulnar nerves between the patients with TTR-FAP and the normal controls ( P>0.05). The side-to-side difference ratio of intra-nerve CSA variability of the ulnar nerve in the patients with TTR-FAP was smaller than that of the normal controls (1.15±0.10 vs 1.46±0.43, t=3.43, P=0.002), whereas no statistically significant difference of that in the median nerve was found between the two groups ( P>0.05). Conclusions:The most pronounced peripheral nerve thickening in the proximal limb segments with no signs of asymmetric distribution or lateralization is confirmed by HFUS in TTR-FAP patients and should be regarded as a marker of TTR-FAP. HFUS has clinical value in diagnosis of peripheral neuropathy in TTR-FAP patients.

Objective: To analyze the epidemiological characteristics of HIV/AIDS infected students in Shandong Province, to provide a basis for the prevention and control of AIDS transmission in the student population. Methods: All 863 HIV/AIDS students cases during 2010-2019 were collected in Shandong Province. Epidemiological characteristics was described and the trends in the 10 years since 2010 was analyzed. Results: These 863 HIV/AIDS students were mainly transmitted through homosexual sex (763 cases, 88.41%), and the samples were mainly from voluntary consultation testing (433 cases, 50.17%). From 2010 to 2019, the proportion of student cases in the total number of cases showed an increasing trend ( χ 2 trend =30.21, P <0.01). Among them, the proportion of homosexual transmission cases increased year by year ( χ 2 trend =6.35, P =0.01), the proportion of cases aged 18-22 years increased year by year ( χ 2 trend =6.10, P =0.01), the proportion of cases with college degree or above increased year by year ( χ 2 trend =4.26, P =0.04). At present, voluntary consultation testing were the main source.There was no significant difference between the years of sample sources ( χ 2 trend =2.97, P =0.09). Conclusion The report number of students in Shandong Province are on the rise in recent years, especially those infected by same sex transmission, mainly with high education background, which calls for targeted strategies and intervention measures.

Objective:To investigate the variation of CD4 + /CD8 + ratio and related influencing factors after receiving antiviral treatment for HIV/AIDS in Shandong province. Methods:HIV/AIDS patients in Shandong province initially received HAART from 2010 to 2015, at least 15 years old with the CD4 + T and CD8 + T cell count tested at baseline, 6 months, 1 year, 2 years, 3 years, 4 years and 5 years after treatment initiation were selected. According to their retrospectively collected baseline and follow-up data, the variation of CD4 + /CD8 + and the related influencing factors were described and analyzed by using rank sum test and generalized estimating equation(GEE). Results:A total of 557 subjects were included in the study. The median baseline CD4 + /CD8 + ratio was 0.20, and the CD4 + /CD8 + ratio was inverted in 99.82% of the patients. The proportion of patients with CD4 + /CD8 + ratio > 1 was 0.18% at baseline, and increased to 13.11% after 5 years of treatment. The trend of time change was statistically significant ( χ2 =95.01, P< 0.05). The result of GEE analysis showed that baseline CD4 + T lymphocytes count level and baseline symptoms/signs were the main factors influencing the variation trend of CD4 + /CD8 + ratio. Conclusions:After 5 years of antiviral treatment with HIV/AIDS in Shandong province, the CD4 + /CD8 + ratio increased significantly. Baseline CD4 + T cell count level and baseline symptoms/signs of patients are the main factors affecting the rate of improvement of CD4 + /CD8 + ratio.

Objective: To analyze the epidemic features of male HIV-infected and AIDS patients by sexual transmission in Shandong Province. Methods: Data on HIV-infected people and AIDS patients (HIV/AIDS) were derived from HIV/AIDS Comprehensive Response Information Management System. To analysis the epidemiological data of male HIV/AIDS by sexual transmission reported in Shandong Province from 1997 to 2016. Results: A total of 8 584 HIV/AIDS were reported by heterosexual transmission or homosexual transmission from 2007 to 2016. 2 421 cases were reported by heterosexual transmission and 6 163 cases were reported by homosexual transmission. Among cases infected by heterosexual transmission. The average age of cases infected by heterosexual transmission was (38.13±12.39) and (31.62±10.22) among cases who infected by homosexual transmission (t=24.95, P<0.001). 84 cases were reported by homosexual transmission and 138 cases by heterosexual transmission from 2007 to 2008, and 6 079 cases were reported by homosexual transmission and 2 283 cases by heterosexual transmission from 2009 to 2016. A total of 770 cases were dead after reported. Among the dead cases, 337 cases were infected by homosexual transmission and 433 cases by heterosexual transmission (χ²=328.21, P<0.001). 61.4% of the dead cases by heterosexual transmission were no longer than 6 months after reported and 54.3% in homosexual transmission (χ²=3.96, P=0.047). Conclusion Homosexual transmission has been the main transmission of HIV/AIDS in Shandong Province. Epidemiological features and social demographic characteristics of each sexual transmission were different. As part of HIV cases developed to death in 6 months.

Objective To explore the collagen, typeⅠ, α 1 chain ( COL1A1) gene mutation in a family with type 1 osteogenesis imperfect. Methods The medical records and DNA samples of an osteogenesis imperfect patient and her family members were collected, and their DNA sequencing were performed and compared with 50 non-relative healthy control from the same area. Results The proband and her three family members ( father, younger brother, and younger nephew) with clinical features of osteogenesis imperfect as well as prolactinoma were confirmed of COL1A1 gene mutation at the 24th intron with a shear mutation of c. 1669-1 G＞A which was not reported previously. Other family members were genetically normal compared with the normal. Conclusions We found a new COL1A1 gene mutation family and mutation site, but the relationship between osteogenesis imperfect and prolactinoma was unknown.