ObjectiveTo investigate the differences in the regulatory effects of formulas containing Ephedrae Herba and Armeniacae Semen Amarum （Mahuangtang， Sanaotang， and Maxing Shigantang） on thermosensitive transient receptor potential ion channels （thermo TRPs） in the mouse model of asthmatic airway inflammation. MethodsSixty female C57BL/6 mice were allocated into blank， model， dexamethasone （0.75 mg·kg^-1）， Mahuangtang （3.8 g·kg^-1）， Sanaotang （2.8 g·kg^-1）， and Maxing Shigantang （6.6 g·kg^-1） groups （n=10）. The mouse model of asthma was established with ovalbumin （OVA） and treated with normal saline （blank group） or corresponding drugs （10 mL·kg^-1）， once a day， 19-28 days after modeling. The levels of eosinophils （EOS） in peripheral blood and white blood cell （WBC） in bronchoalveolar lavage fluid （BALF）， changes in enhanced pause （Penh）， and pathological damage of lung tissue were observed in each group. Western blot and real-time PCR were employed to quantify the protein and mRNA levels， respectively， of high-temperature thermosensitive channels （TRPV1 and TRPV3） and low-temperature thermosensitive channels （TRPA1 and TRPM8） in the lung tissue. ResultsCompared with the blank group， the model group showed a typical asthma phenotype， including elevations in the level of EOS in peripheral blood， level of WBC in BALF， and value of Penh （P<0.05，P<0.01）， and severe lung tissue damage. Compared with the model group， the three formulas alleviated the asthma phenotype to varying degrees （P<0.05，P<0.01）. Compared with the blank group， the model group showed up-regulated protein levels of TRPV1 and TRPA1 in the lung tissue （P<0.01）. Compared with the model group， Maxing Shigantang and Sanaotang groups showed down-regulated protein levels of TRPV1 and TRPA1 （P<0.05， P<0.01）. Moreover， Maxing Shigantang and Sanaotang groups showed more significant down-regulation in protein levels of TRPV1 and TRPA1， respectively （P<0.01）， while no obvious regulatory effect was observed in the Mahuangtang group. Compared with those in the blank group， the protein levels of TRPV3 and TRPM8 were up-regulated in the model group （P<0.01）. Compared with the model group， Maxing Shigantang and Sanaotang down-regulated the protein levels of TRPV3 and TRPM8 （P<0.01）. Moreover， Maxing Shigantang and Sanaotang exerted stronger down-regulating effects on TRPV3 （P<0.05） and TRPM8 （P<0.01）， respectively. Compared with the blank group， the model group presented up-regulated mRNA levels of TRPV1， TRPV3， TRPA1， and TRPM8 in the lung tissue （P<0.01）， and such up-regulations were significantly decreased by Maxing Shigantang and Sanaotang （P<0.01）. Moreover， Maxing Shigantang outperformed Sanaotang in regulating high-temperature thermosensitive channels TRPV1 and TRPV3 （P<0.05， P<0.01）. The regulation effect of the， Maxing Shigantang on high-temperature thermosensory channel proteins of TRPV1 and TRPV3 was better than that of the Sanaotang P<0.05P<0.01while the Sanaotang outperformedhad a significant regulatory effect on Maxing Shigantang in regulating the low-temperature thermosensory thermosensitive channel proteins of TRPA1 and TRPM8which was better than that of the Maxing Shigantang （P<0.05，P<0.01）. ConclusionThe experimental results showed that Mahuangtang， Sanaotang， and Maxing Shigantang all had protective effects on asthma airway inflammation.while Mahuangtang did not show the regulatory effect on TRPV1 and or TRPA1. Maxing Shigantang preferred to regulate high-temperature thermosensory thermosensitive channels of TRPV1 and TRPV3 channels， and Sanaotang preferred to regulate low-temperature thermosensory thermosensitive channels of TRPA1 and TRPM8.

This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5％.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.

Objective:To evaluate the long-term efficacy and safety of recombinant coagulation factor Ⅷ (Octocog alfa) in Chinese patients with hemophilia A (HA) enrolled in the International Antihemophilic Factor Hemophilia A Outcome Database (AHEAD) study (NCT02078427) .Methods:Enrollment of Chinese patients in the AHEAD study was completed by January 2021, and data were collected up to July 15, 2022. This study primarily assessed patients in terms of the Gilbert score, global gait score within the Hemophilia Joint Health Score (HJHS), annualized bleeding rate (ABR), annualized joint bleeding rate, and adverse events.Results:A total of 168 male patients were included in this study, of which 113 received prophylactic treatment and 53 received on-demand treatment. The average age of the patients was 21.4±13.37 years. Compared with baseline, the global gait score within HJHS significantly decreased during the 1-year follow-up in patients with moderately severe HA in the prophylactic treatment group ( P=0.01) and on-demand treatment group ( P=0.008). The mean reduction in Gilbert score was greater in the prophylactic treatment group than in the on-demand treatment group (28.6% vs 8.2%). The average ABR decreased significantly during the 1-year follow-up (3.70 vs 7.78, P=0.01) in the prophylactic treatment group, particularly in patients with severe HA (2.14 vs 8.98, P=0.006) and pediatric patients (2.1 vs 4.73, P=0.03). The ABR score also decreased significantly in the moderate-dose prophylactic treatment group ( P=0.015). During the 1-year follow-up, 25 patients (14.9%) reported 39 adverse events, with only one patient developing treatment-related F Ⅷ inhibitor. Conclusion:Joint mobility improved in patients receiving either prophylactic or on-demand Octocog alfa. Bleeding episodes significantly reduced in patients receiving prophylactic treatment, particularly in pediatric patients and those with severe HA.

The anti-inflammatory phytochemical investigation of the leaves of Illicium dunnianum (I. dunnianum) resulted in the isolation of five pairs of new lignans (1-5), and 7 known analogs (6-12). The separation of enantiomer mixtures 1-5 to 1a/1b-5a/5b was achieved using a chiral column with acetonitrile-water mixtures as eluents. The planar structures of 1-2 were previously undescribed, and the chiral separation and absolute configurations of 3-5 were reported for the first time. Their structures were determined through comprehensive spectroscopic data analysis [nuclear magnetic resonance (NMR), high-resolution electrospray ionization mass (HR-ESI-MS), infrared (IR), and ultraviolet (UV)] and quantum chemistry calculations (ECD). The new isolates were evaluated by measuring their inhibitory effect on NO in lipopolysaccharide (LPS)-stimulated BV-2 cells. Compounds 1a, 3a, 3b, and 5a demonstrated partial inhibition of NO production in a concentration-dependent manner. Western blot and real-time polymerase chain reaction (PCR) assays revealed that 1a down-regulated the messenger ribonucleic acid (mRNA) levels of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), COX-2, and iNOS and the protein expressions of COX-2 and iNOS. This research provides guidance and evidence for the further development and utilization of I. dunnianum.
Lignans/isolation & purification* ; Plant Leaves/chemistry* ; Anti-Inflammatory Agents/isolation & purification* ; Mice ; Animals ; Molecular Structure ; Plant Extracts/pharmacology* ; Illicium/chemistry* ; Cyclooxygenase 2/immunology* ; Interleukin-6/immunology* ; Nitric Oxide/metabolism* ; Cell Line ; Tumor Necrosis Factor-alpha/immunology* ; Nitric Oxide Synthase Type II/immunology* ; Lipopolysaccharides

Objective:To observe the effects of 12.5% carbohydrate (CHO) solution and medium chain triglycerides (MCT) solutions at various concentrations on gastric emptying, in order to determine the optimal MCT-CHO combination for enhanced recovery after surgery.Methods:This study was a prospective study. Ten healthy volunteers were selected to ingest 400 ml of the following 5 solutions every day: water, 12.5% CHO, 1% MCT, 2% MCT and 4% MCT. According to the above results, the optimal concentration of MCT solution was 2% MCT, showing comparable gastric emptying rate compared with 12.5% CHO solution. On this basis, combinations of MCT and CHO solutions were determined to be 30 energy % (EN%) MCT+70 EN% CHO and 20 EN% MCT+80 EN% CHO according to the energy distribution ratio. Then the volunteers were given 400 ml of the following 4 solutions every day: 12.5% CHO, 2% MCT, 30 EN% MCT+70 EN% CHO (equivalent to 2% MCT+8% CHO), and 20 EN% MCT+80 EN% CHO (equivalent to 4.4% MCT+10% CHO). Gastric emptying during fasting (T m) and immediately (T 1), 30 min (T 2), 60 min (T 3), 90 min (T 4) and 120 min after ingestion (T 5) were observed by antral ultrasonography. The degree of thirst, hunger, and anxiety was assessed by Visual Analogue Scale, and the taste was rated. Results:The gastric emptying rate in descending order was 20%EN MCT+80%EN CHO group>30%EN MCT+70%EN CHO group≈2％MCT group≈12.5%CHO group. There was no significant difference in gastric emptying rate between 30 EN% MCT+70 EN% CHO group and 12.5% CHO group ( P>0.05). The scores of thirst, hunger and anxiety in 30 EN% MCT+70 EN% CHO group at T 1 to T 5 were significantly lower than those at T m ( P<0.05). 30 EN% MCT+70 EN% CHO group showed the highest taste score. Conclusion:30 EN% MCT+70 EN% CHO solution has similar gastric emptying rate compared with 12.5% CHO solution. It can relieve thirst, hunger, anxiety and other subjective feelings caused by fasting, with tastes better than 12.5% CHO solution.

To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.

To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.

Objective:To observe the effects of 12.5% carbohydrate (CHO) solution and medium chain triglycerides (MCT) solutions at various concentrations on gastric emptying, in order to determine the optimal MCT-CHO combination for enhanced recovery after surgery.Methods:This study was a prospective study. Ten healthy volunteers were selected to ingest 400 ml of the following 5 solutions every day: water, 12.5% CHO, 1% MCT, 2% MCT and 4% MCT. According to the above results, the optimal concentration of MCT solution was 2% MCT, showing comparable gastric emptying rate compared with 12.5% CHO solution. On this basis, combinations of MCT and CHO solutions were determined to be 30 energy % (EN%) MCT+70 EN% CHO and 20 EN% MCT+80 EN% CHO according to the energy distribution ratio. Then the volunteers were given 400 ml of the following 4 solutions every day: 12.5% CHO, 2% MCT, 30 EN% MCT+70 EN% CHO (equivalent to 2% MCT+8% CHO), and 20 EN% MCT+80 EN% CHO (equivalent to 4.4% MCT+10% CHO). Gastric emptying during fasting (T m) and immediately (T 1), 30 min (T 2), 60 min (T 3), 90 min (T 4) and 120 min after ingestion (T 5) were observed by antral ultrasonography. The degree of thirst, hunger, and anxiety was assessed by Visual Analogue Scale, and the taste was rated. Results:The gastric emptying rate in descending order was 20%EN MCT+80%EN CHO group>30%EN MCT+70%EN CHO group≈2％MCT group≈12.5%CHO group. There was no significant difference in gastric emptying rate between 30 EN% MCT+70 EN% CHO group and 12.5% CHO group ( P>0.05). The scores of thirst, hunger and anxiety in 30 EN% MCT+70 EN% CHO group at T 1 to T 5 were significantly lower than those at T m ( P<0.05). 30 EN% MCT+70 EN% CHO group showed the highest taste score. Conclusion:30 EN% MCT+70 EN% CHO solution has similar gastric emptying rate compared with 12.5% CHO solution. It can relieve thirst, hunger, anxiety and other subjective feelings caused by fasting, with tastes better than 12.5% CHO solution.

This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5％.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.

Objective:To evaluate the long-term efficacy and safety of recombinant coagulation factor Ⅷ (Octocog alfa) in Chinese patients with hemophilia A (HA) enrolled in the International Antihemophilic Factor Hemophilia A Outcome Database (AHEAD) study (NCT02078427) .Methods:Enrollment of Chinese patients in the AHEAD study was completed by January 2021, and data were collected up to July 15, 2022. This study primarily assessed patients in terms of the Gilbert score, global gait score within the Hemophilia Joint Health Score (HJHS), annualized bleeding rate (ABR), annualized joint bleeding rate, and adverse events.Results:A total of 168 male patients were included in this study, of which 113 received prophylactic treatment and 53 received on-demand treatment. The average age of the patients was 21.4±13.37 years. Compared with baseline, the global gait score within HJHS significantly decreased during the 1-year follow-up in patients with moderately severe HA in the prophylactic treatment group ( P=0.01) and on-demand treatment group ( P=0.008). The mean reduction in Gilbert score was greater in the prophylactic treatment group than in the on-demand treatment group (28.6% vs 8.2%). The average ABR decreased significantly during the 1-year follow-up (3.70 vs 7.78, P=0.01) in the prophylactic treatment group, particularly in patients with severe HA (2.14 vs 8.98, P=0.006) and pediatric patients (2.1 vs 4.73, P=0.03). The ABR score also decreased significantly in the moderate-dose prophylactic treatment group ( P=0.015). During the 1-year follow-up, 25 patients (14.9%) reported 39 adverse events, with only one patient developing treatment-related F Ⅷ inhibitor. Conclusion:Joint mobility improved in patients receiving either prophylactic or on-demand Octocog alfa. Bleeding episodes significantly reduced in patients receiving prophylactic treatment, particularly in pediatric patients and those with severe HA.