Calcitonin gene-related peptides(CGRP)are peptides that are ubiquitous in the central and peripheral nervous systems,which has been shown to be associated with pain,setting off a boom in the development of chemical small molecule antagonists targeting CGRP receptors,but most of them have failed due to greater hepatotoxicity.Anti-CGRP/CGRPR antibodies have attracted widespread attention due to their high specificity and safety,especially lower hepatotoxicity.This article reviews the structure and function of CGRP and its receptors,as well as the research progress of anti-CGRP-related neutralizing antibodies in order to provide a reference for the exploration of disease mechanisms and drug development related to CGRP.

Calcitonin gene-related peptides(CGRP)are peptides that are ubiquitous in the central and peripheral nervous systems,which has been shown to be associated with pain,setting off a boom in the development of chemical small molecule antagonists targeting CGRP receptors,but most of them have failed due to greater hepatotoxicity.Anti-CGRP/CGRPR antibodies have attracted widespread attention due to their high specificity and safety,especially lower hepatotoxicity.This article reviews the structure and function of CGRP and its receptors,as well as the research progress of anti-CGRP-related neutralizing antibodies in order to provide a reference for the exploration of disease mechanisms and drug development related to CGRP.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore the relationship between the clinical outcome of emotional symptoms and cognitive performance and related cerebral oxygenated hemoglobin in adolescents with depression.Methods:Through subject recruitment, 46 adolescent patients with depression (patient group) from the First Hospital of Shanxi Medical University were selected as the subjects for this study from December 2020 to December 2021, including 8 males and 38 females, aged 12-18 (15.7±2.3) years old. All patients received sertraline treatment for 8 weeks and were further followed into responders ( n=24) and non-responders ( n=22) according to the outcome of emotional symptoms. In the meantime, 51 healthy controls (control group) were enrolled, including 7 males and 44 females, aged 12-18 (16.1±1.5) years old. The repeatable battery for the assessment of neuropsychological status (RBANS) was conducted to measure the multi-dimensional neurocognitive performance, and the functional near-infrared spectroscopy (fNIRS) was used to assess changes in the concentration of oxyhemoglobin (HBO) during the verbal fluency test. The differences were compared in multi-dimensional cognitive performance and cerebral HBO level between each patient group and control group and between responders and non-responders. The changes were analyzed in cognitive performance and cerebral HBO level after intervention in responders and non-responders. Results:At baseline, compared to the control group, the patient group performed decreased scores of RBANS, immediate memory, speech function, attention, and delayed memory (88.0 (82.8, 100.0) M ( Q1, Q3) vs. 100.0 (90.0, 110.0) scores; 78.0 (73.0, 87.8) vs.85.0 (78.0, 94.0) scores; (84.4±16.1) vs. (95.7±15.7) scores; 106.0 (99.5, 115.0) vs.118.0 (109.0, 128.0) scores; 94.0 (84.5, 99.0) vs.97.0 (91.0, 101.0) scores), and lower HBO levels in 7 channels (all P<0.05). Compared to responders, non-responders showed more severe impairment of visual-spatial and attention performance (103.9±11.0 vs. 94.4±16.7 scores; 112.5±12.1 vs. 98.0±21.2 scores) ( t=2.30 or 2.87; all P<0.05). After treatment, the scores of RBANS and immediate memory improved significantly in responders (98.8±11.2 vs. 93.0±9.7 scores; 95.2±13.8 vs.83.0±14.6 scores) ( t=-3.00 or-4.97; both P<0.05), but the scores of attention and the HBO level of two channels in the prefrontal cortex were still significantly lower than those of the control group (Z=2.27, 3.02 or 3.04; all P<0.05). Besides, there were no significant differences in the scores of immediate memory and the HBO levels of 3 channels in the temporal lobe between the no-responders and the control group (all P>0.05). Conclusion:Immediate memory injure, attention injure and HBO levels of frontal-temporal lobes may be independent of emotional symptoms among adolescents with depression.

Objective:To establish an antibody expression system to reduce the antibody-dependent enhancement (ADE) effect of target antibody.Methods:Site-directed mutagenesis was used to mutate the 234 and 235 sites of the Fc region of the mammalian cell antibody expression vector-L234A and L235A to establish the antibody expression vector pFRT-IgG1κ-FcM. An antibody Wt-WNV with significant ADE effect obtained in previous work was selected and expressed by the pFRT-IgG1κ-FcM system to obtain mutant antibody FcM-WNV. The binding ability of FcM-WNV to target antigen West Nile virus envelope protein-DⅢ (WNV E-DⅢ) was detected by ELISA, and the its binding ability to human high-affinity IgG Fc receptor hFcγRⅠ (hCD64 ) was analyzed by flow cytometry. The neutralizing activity of FcM-WNV in vitro was detected by pseudovirus infection of host cells (BHK21 and K562). Results:The expression levels of FcM-WNV and Wt-WNV were comparable, and FcM-WNV could recognize and bind to WNVE-DIII in a concentration-dependent manner. Compared with Wt-WNV, the binding ability of FcM-WNV to hCD64 was significantly weakened, showing a significant decrease in fluorescence intensity. Consistent with the previous experimental results, Wt-WNV at a concentration of 5 μg/ml significantly enhanced the infection of K562 by WNV pseudovirus, while FcM-WNV at a concentration of 5 μg/ml could effectively block pseudovirus infection in both K562 and BHK21 cells.Conclusions:The established antibody expression system can effectively reduce the ADE effect of the target antibody.

Objective:To establish an in vivo infection model of West Nile virus (WNV) pseudovirus and evaluate the neutralizing activity of antibody WNV-XH1.Methods:A stable cell line that can package the WNV pseudovirus was established in the early stage to prepare the pseudovirus supernatant. The supernatant was concentrated and infected BHK21 cells to detect the titer of the pseudovirus. After intraperitoneal injection of the pseudovirus into C57BL/J mice, bioluminescence imaging was performed to observe the infection status of the pseudovirus in the mice. After simultaneous infection, blood was collected and ELISA was used to detect NS1 levels in mouse serum. The in vivo functional activity of antibody WNV-XH1 was evaluated using the established mouse infection model.Results:Fluorescence was detected in C57BL/J mice infected with WNV pseudovirus, and the NS1 levels in the peripheral blood serum of mice infected with pseudovirus were significantly higher than those of non infected mice (1.453±0.09vs0.305±0.018). After intravenous administration of WNV-XH1 antibody before the attack, the fluorescence signal in the mice decreased and the serum NS1 level decreased (0.384±0.015).Conclusions:A successful in vivo infection model of WNV pseudovirus was established, and it was confirmed that the antibody WNV-XH1 had a protective effect against WNV pseudovirus infection in vivo.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective:To explore the relationship between the clinical outcome of emotional symptoms and cognitive performance and related cerebral oxygenated hemoglobin in adolescents with depression.Methods:Through subject recruitment, 46 adolescent patients with depression (patient group) from the First Hospital of Shanxi Medical University were selected as the subjects for this study from December 2020 to December 2021, including 8 males and 38 females, aged 12-18 (15.7±2.3) years old. All patients received sertraline treatment for 8 weeks and were further followed into responders ( n=24) and non-responders ( n=22) according to the outcome of emotional symptoms. In the meantime, 51 healthy controls (control group) were enrolled, including 7 males and 44 females, aged 12-18 (16.1±1.5) years old. The repeatable battery for the assessment of neuropsychological status (RBANS) was conducted to measure the multi-dimensional neurocognitive performance, and the functional near-infrared spectroscopy (fNIRS) was used to assess changes in the concentration of oxyhemoglobin (HBO) during the verbal fluency test. The differences were compared in multi-dimensional cognitive performance and cerebral HBO level between each patient group and control group and between responders and non-responders. The changes were analyzed in cognitive performance and cerebral HBO level after intervention in responders and non-responders. Results:At baseline, compared to the control group, the patient group performed decreased scores of RBANS, immediate memory, speech function, attention, and delayed memory (88.0 (82.8, 100.0) M ( Q1, Q3) vs. 100.0 (90.0, 110.0) scores; 78.0 (73.0, 87.8) vs.85.0 (78.0, 94.0) scores; (84.4±16.1) vs. (95.7±15.7) scores; 106.0 (99.5, 115.0) vs.118.0 (109.0, 128.0) scores; 94.0 (84.5, 99.0) vs.97.0 (91.0, 101.0) scores), and lower HBO levels in 7 channels (all P<0.05). Compared to responders, non-responders showed more severe impairment of visual-spatial and attention performance (103.9±11.0 vs. 94.4±16.7 scores; 112.5±12.1 vs. 98.0±21.2 scores) ( t=2.30 or 2.87; all P<0.05). After treatment, the scores of RBANS and immediate memory improved significantly in responders (98.8±11.2 vs. 93.0±9.7 scores; 95.2±13.8 vs.83.0±14.6 scores) ( t=-3.00 or-4.97; both P<0.05), but the scores of attention and the HBO level of two channels in the prefrontal cortex were still significantly lower than those of the control group (Z=2.27, 3.02 or 3.04; all P<0.05). Besides, there were no significant differences in the scores of immediate memory and the HBO levels of 3 channels in the temporal lobe between the no-responders and the control group (all P>0.05). Conclusion:Immediate memory injure, attention injure and HBO levels of frontal-temporal lobes may be independent of emotional symptoms among adolescents with depression.

Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.