OBJECTIVE To screen and obtain nanobodies with neutralizing activity against granulo-cyte-macrophage colony stimulating factor(GM-CSF)to contribute to investigations into the mecha-nism of inflammation interventions and the development new drugs.METHODS Recombinant human GM-CSF was subcutaneously injected to immunize camels.Peripheral blood was collected after five immunizations,and mononuclear cells were isolated.Total mRNA was extracted,and the variable domains of the heavy chain of heavy-chain antibody(VHH,also called nanobody)genes were obtained by PCR amplification after reverse transcription.The genes were cloned into the pADSCFV-S phage display vector and electrotransformed into TG1 competent cells to construct a nanobody immune library that was screened with recombinant human GM-CSF immobilized on a solid phase.The VHH genes specifi-cally binding to human GM-CSF were cloned into the pABG eukaryotic expression vector before VHH-Fc samples were prepared by using the human embryonic kidney 293 fibroblast expression system.The binding activity of candidate VHH-Fc molecules to GM-CSF was investigated through ELISA response curves,and binding colorimetric values with different antigens were detected to determine their specificity.The binding affinity between VHH-Fc candidates and GM-CSF was measured using biolayer interferom-etry(BLI).The inhibition rate curve of VHH-Fc candidates on GM-CSF was detected through cell prolif-eration assays to determine its neutralization activity.The Uncle system was used to investigate its thermal stability.100 μg of VHH-Fc was injected into mice via the tail vein,and the serum concentration of VHH-Fc was quantitatively detected by ELISA to examine its pharmacokinetic curve in mice.RESULTS The camel serum titer of anti-human GM-CSF antibodies was higher than 1:800 000 after the fifth immuni-zation,and the capacity of the constructed nanobody library was about 5.55×107.Following the screening process,five candidate VHH-Fc molecules specifically binding to human GM-CSF were obtained.Among these,22N10 effectively neutralized the cell proliferation-promoting activity of GM-CSF(the IC50 value was 17.23 nmol·L-1).Subsequent studies revealed that 22N10 interacted with human GM-CSF with an affinity of 1.97×10-8 mol·L-1,blocked the binding of GM-CSF to its receptor CSF2Rα,exhibited good thermal stability(Tm1=59.2℃),and showed favorable metabolic characteristics in mice.CONCLU-SION A new candidate nanobody molecule 22N10 targeting human GM-CSF is obtained which is expected to facilitate the drug development and mechanism investigations.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Point-of-care testing(POCT)is a new model of medical testing development,which develops rapidly in recent years.POCT has the characteristics of rapid detection,convenient operation,and can be car-ried out at the patient's bedside.It can provide favorable conditions for early diagnosis of patients with critical illnesses in the emergency department and can significantly reduce their consultation time,alleviate the over-crowding of the emergency department,which is appropriate to the needs of the development of emergency medicine.This paper mainly introduces the specific application and research progress of POCT in emergency medicine,analyzes the advantages and challenges of POCT technology,and discusses the new technology and development prospect of POCT.

OBJECTIVE To screen and obtain nanobodies with neutralizing activity against granulo-cyte-macrophage colony stimulating factor(GM-CSF)to contribute to investigations into the mecha-nism of inflammation interventions and the development new drugs.METHODS Recombinant human GM-CSF was subcutaneously injected to immunize camels.Peripheral blood was collected after five immunizations,and mononuclear cells were isolated.Total mRNA was extracted,and the variable domains of the heavy chain of heavy-chain antibody(VHH,also called nanobody)genes were obtained by PCR amplification after reverse transcription.The genes were cloned into the pADSCFV-S phage display vector and electrotransformed into TG1 competent cells to construct a nanobody immune library that was screened with recombinant human GM-CSF immobilized on a solid phase.The VHH genes specifi-cally binding to human GM-CSF were cloned into the pABG eukaryotic expression vector before VHH-Fc samples were prepared by using the human embryonic kidney 293 fibroblast expression system.The binding activity of candidate VHH-Fc molecules to GM-CSF was investigated through ELISA response curves,and binding colorimetric values with different antigens were detected to determine their specificity.The binding affinity between VHH-Fc candidates and GM-CSF was measured using biolayer interferom-etry(BLI).The inhibition rate curve of VHH-Fc candidates on GM-CSF was detected through cell prolif-eration assays to determine its neutralization activity.The Uncle system was used to investigate its thermal stability.100 μg of VHH-Fc was injected into mice via the tail vein,and the serum concentration of VHH-Fc was quantitatively detected by ELISA to examine its pharmacokinetic curve in mice.RESULTS The camel serum titer of anti-human GM-CSF antibodies was higher than 1:800 000 after the fifth immuni-zation,and the capacity of the constructed nanobody library was about 5.55×107.Following the screening process,five candidate VHH-Fc molecules specifically binding to human GM-CSF were obtained.Among these,22N10 effectively neutralized the cell proliferation-promoting activity of GM-CSF(the IC50 value was 17.23 nmol·L-1).Subsequent studies revealed that 22N10 interacted with human GM-CSF with an affinity of 1.97×10-8 mol·L-1,blocked the binding of GM-CSF to its receptor CSF2Rα,exhibited good thermal stability(Tm1=59.2℃),and showed favorable metabolic characteristics in mice.CONCLU-SION A new candidate nanobody molecule 22N10 targeting human GM-CSF is obtained which is expected to facilitate the drug development and mechanism investigations.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective To investigate the correlation between red blood cell distribution width(RDW),systemic immune-inflammation index(SII)and major depressive disorder(MDD).Methods The clinical data of 176 MDD patients hospitalized in the clinical psychology department of our hospital from 2020 to 2022 and 209 non-MDD comparators who were routinely examined in Hebei General Hospital were retrospectively analyzed.Blood analysis was conducted to obtain values of RDW,SII,and red blood cell distribution width/platelet ratio(RPR).The data was used to plot the receiver operator characteristic(ROC)curve to determine the optimal threshold and the area under the curve(AUC)for RDW to discriminate between patients and controls.Result Patients in the MDD group had significantly higher RDW[median and quartiles:13.20(12.70,13.98)vs.12.80(12.40,13.35)],and SII levels[median and quartiles 510.87(350.95,878.12)vs.405.33(313.74,539.92)]compared with those in non-MDD group(P<0.05).There was no significant difference in RPR between the two groups(P>0.05).Multifactorial logistic regression analysis showed that RDW was positively correlated with MDD after adjusting for confounders(OR=3.086,95% CI:1.926-4.944).ROC curve showed that the optimal threshold for RDW to differentiate the risk of developing MDD was 12.85,with an AUC of 0.647(95% CI:0.592-0.702;P<0.001).Conclusion Present study shows that high RDW is a risk factor for the occurrence of MDD and an important parameter for the risk of developing depression.

Objective:To investigate the efficacy of dapagliflozin combined with tirofiban in patients with type 2 diabetes mellitus complicated with coronary heart disease.Methods:A total of 120 patients with type 2 diabetes mellitus and coronary heart disease treated in Fuyang People′s Hospital from January to August 2022 were prospectively selected as subjects. According to different treatment methods, they were divided into control group and experimental group. The control group was treated with tirofiban, and the experimental group was treated with dapagliflozin combined with tirofiban. The efficacy and safety of treatments between the two groups were compared.Results:After 3 months of treatment, fasting plasma glucose (FPG), 2 h postprandional blood glucose (2 h PG), glycated hemoglobin (HbA 1c), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), D-dimer (D-D) and fibrinogen (FIB) levels in 2 groups were decreased compared with before treatment:experimental group: (7.33 ± 1.77) mmol/L vs. (9.45 ± 2.05) mmol/L, (10.33 ± 2.07) mmol/L vs. (13.57 ± 2.88) mmol/L, (7.22 ± 1.28) % vs. (9.25 ± 1.78) %, (1.98 ± 0.29) mmol/L vs. (6.05 ± 1.24) mmol/L, (2.95 ± 0.37) mmol/L vs. (4.35 ± 0.76) mmol/L, (0.78 ± 0.23) mg/L vs. (1.85 ± 0.79) mg/L, (2.57 ± 0.37) g/L vs. (7.15 ± 1.36) g/L, control group: (8.21 ± 1.85) mmol/L vs. (9.68 ± 2.17) mmol/L, (11.78 ± 2.26) mmol/L vs. (13.89 ± 3.02) mmol/L, (8.25 ± 1.36) % vs. (9.37 ± 1.86) %, (2.77 ± 0.42) mmol/L vs. (6.08 ± 1.22) mmol/L, (3.84 ± 0.44) mmol/L vs. (4.27 ± 0.72) mmol/L, (1.34 ± 0.52) mg/L vs. (1.81 ± 0.72) mg/L, (5.25 ± 0.84 ) g/L vs. (7.11 ± 1.28) g/L, the differences were statistically significant ( P< 0.05). The levels of FPG, 2 h PG, HbA 1c, TC, LDL-C, D-D and FIB between the two groups were statistically significant ( P<0.05). High density lipoprotein cholesterol (HDL-C) level, left ventricular ejection fraction (LVEF), cardiac blood transfusion volume (CO), stroke output (SV), thrombin time (TT) and partially activated prothrombin time (APTT) were all increased: experimental group: (1.76 ± 0.30) mmol/L vs. (1.07 ± 0.28) mmol/L, (68.64 ± 12.91) % vs. (45.05 ± 12.24) %, (4.88 ± 0.91) L/min vs. (3.95 ± 1.12) L/min, (53.66 ± 5.43) ml/min vs. (43.27 ± 4.88) ml/min, (31.83 ± 4.39) s vs. (23.25 ± 3.44) s, (13.85 ± 2.17) s vs. (10.75 ± 1.56) s, control group: (1.43 ± 0.26) mmol/L vs. (1.06 ± 0.24) mmol/L, (60.37 ± 11.86) % vs. (45.42 ± 12.41) %, (4.37 ± 0.84) L/min vs. (4.17 ± 1.16) L/min, (47.86 ± 5.27) ml/min vs. (43.36 ± 4.94) ml/min, (27.24 ± 3.91) s vs. (23.78 ± 3.62) s, (12.74 ± 1.94) s vs. (10.89 ± 1.78) s, the differences were statistically significant ( P<0.05). There were significant differences in HDL-C, LVEF, CO, SV, TT and APTT between the two groups ( P<0.05). The incidence of adverse reactions in experimental group was lower than that in control group during treatment: 5.00% (3/60) vs. 16.67% (10/60), and the difference was statistically significant ( P<0.05). Conclusions:Dapagliflozin combined with tirofiban in the treatment of patients with type 2 diabetes mellitus complicated with coronary heart disease has obvious curative effect, and can improve blood glucose and blood lipid levels, coagulation function and cardiac function, with high safety.

Objective:To evaluate the clinical effects of transperitoneal and retroperitoneal laparoscopic decortication in the treatment of simple renal cysts.Methods:PubMed, EMBASE, Cochrane Library, CNKI, VIP and WanFang databases were searched, All studies about comparing transperitoneal and retroperitoneal laparoscopic decortication for simple renal cysts were included. The evaluation indicators included operative time, intraoperative blood loss, postoperative drainage time, postoperative hospital stay, and complication rate. Statistical analysis of the data using RevMan 5.3 software.Results:A total of 14 control studies were included, 409 cases in the transperitoneal group and 452 cases in the retroperitoneal group. Meta-analysis demonstrated that compared with the transperitoneal group, the retroperitoneal group had shorter operation time ( MD=8.81, 95% CI: 3.79-13.82, P<0.01), less intraoperative blood loss ( MD=4.89, 95% CI: 1.40-8.38, P<0.01), and shorter postoperative hospital stay ( MD=0.60, 95% CI: 0.24-0.96, P<0.01). However, there were not significant differences between the two approaches in postoperative drainage time ( MD=-0.12, 95% CI: -0.26-0.03, P=0.13) and complication rate ( OR=1.47, 95% CI: 0.48-4.47, P=0.05). Conclusions:Compared with the transperitoneal, the retroperitoneal laparoscopic decortication has the advantages of short operation time, less intraoperative blood loss and short postoperative hospital stay. Therefore, retroperitoneal laparoscopic decortication is expected to be the first choice for the treatment of simple renal cysts.

A neuroinflammatory response is commonly involved in the progression of many neurodegenerative diseases. Potassium 2-(1-hydroxypentyl)-benzoate (PHPB), a novel neuroprotective compound, has shown promising effects in the treatment of ischemic stroke and Alzheimer׳s disease (AD). In the present study, the anti-inflammatory effects of PHPB were investigated in the plasma and brain of C57BL/6 mice administered a single intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). Levels of iNOS and the cytokines TNF, IL-1and IL-10 were elevated in plasma, cerebral cortex and hippocampus after LPS injection and the number of microglia and astrocytes in cortex and hippocampus were increased. LPS also upregulated the expression of heme oxygenase-1 (HO-1) in the cortex and hippocampus. PHPB reduced the levels of iNOS and cytokines in the plasma and brain, decreased the number of microglia and astrocytes and further enhanced the upregulation of HO-1. In addition, PHPB inhibited the LPS-induced phosphorylation of ERK, P38 and JNK. These results suggest that PHPB is a potential candidate in the treatment of neurodegenerative diseases through inhibiting neuroinflammation.

Objective To investigate the clinical value of arterial first approach in laparoscopic pancreaticoduodenectomy (LPD).Methods The retrospective cohort study was conducted.The clinicopathological data of 181 patients with pancreatic head and periampullay tumors who underwent LPD in the Affiliated Tongji Hospital of Huazhong University of Science and Technology between October 2014 and December 2016 were collected.Among 181 patients,96 using arterial first approach and 85 using traditional approach were respectively allocated into the experimental group and the control group.Surgery was applied to patients in the same doctors' team,and there were the same extent of surgical resection,range of lymph node dissection and digestive tract reconstruction.Observation indicators:(1) intraoperative situation;(2) postoperative situation;(3) followup and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect the tumor-free survival up to February 2017.Measurement data with normal distribution were represented as x±s,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the chi-square test or Fisher exact probability.Results (1) Intraoperative situation:all the patients underwent successful LPD.Overall operation time and time of digestive tract reconstruction were respectively (268 ± 20) minutes,(33 ± 10) minutes in the experimental group and (285±25)minutes,(30± 17)minutes in the control group,with no statistically significant difference between 2 groups (t =8.529,2.741,P＞ 0.05).Time of tumor resection with superior mesenteric venous invasion were respectively (216± 13)minutes and (264±22)minutes in the experimental and control groups,with a statistically significant difference between the 2 groups (t=41.826,P＜0.05).Time of tumor resection without superior mesenteric venous invasion were respectively (224± 14) minutes and (215±21) minutes in the experimental and control groups,with no statistically significant difference between the 2 groups (t =7.423,P＞ 0.05).Volumes of intraoperative blood loss and blood transfusion were respectively (99± 16)mL,(1.3±0.8)U in the experimental group and (131±27)mL,(2.8±1.2)U in the control group,with statistically significant differences between the 2 groups (t =3.670,0.562,P＜ 0.05).Five and 8 patients had intraoperative blood transfusion in the experimental and control groups,showing no statistically significant difference between the 2 groups (x2=1.195,P＞0.05).(2) Postoperative situation:time of drainage tube removal and duration of hospital stay were respectively (5.8±2.4)days,(18.3±6.3) days in the experimental group and (6.3±3.6)days,(19.6±7.1) days in the control group,with no statistically significant difference between the 2 groups (t =0.498,1.305,P＞0.05).Eleven patients in the experimental group had postoperative early complications,including 8with grade A pancreatic fistula (4 combined with diarrhea,2 combined with biliary fistula,1 combined with delayed gastric emptying and 1 with single pancreatic fistula),3 with grade B pancreatic fistula (2 combined with intra-abdominal hemorrhage and 1 combined with intra-abdominal infection).One patient with intra-abdominal hemorrhage in the experimental group died after treatment failure.Twelve patients in the control group had postoperative early complications,including 6 with grade A pancreatic fistula (2 combined with biliary fistula,2 combined with delayed gastric emptying,1 combined with diarrhea,1 combined with digestive tract hemorrhage),3 with grade B pancreatic fistula and intra-abdominal hemorrhage (2 combined with infection,including 1 death) and 3 with diarrhea.Other patients with complications were cured by symptomatic and supportive treatment.There was no statistically significant difference in overall complications between the 2 groups (x2 =0.287,P＞0.05).Results of postoperative pathological examination showed that case with R0 resection was 93 and 76 in the experimental and control groups,with a statistically significant difference between the 2 groups (x2 =4.057,P＜0.05).(3) Follow-up and survival situations:179 patients were followed up for 2-28 months,with a median time of 14 months.Postoperative 6-month tumor-free survival rate was 92.7％ (89/96) and 88.2％(75/85) in the experimental and control groups,with no statistically significant difference between the 2 groups (x2=1.060,P＞0.05).Conclusion Arterial first approach in LPD could significantly shorten the time of tumor resection of patients with superior mesenteric artery invading pancreatic head and periampullay region,significantly reduce the volumes of intraoperative blood loss and blood transfusion,and increase the rate of R0 resection.