To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objectives:To investigate the imaging factors influencing the early prognosis of syrinx in patients with Chiari malformation type Ⅰ(CM Ⅰ)and syringomyelia(SM)who underwent posterior fossa decompression(PFD).Methods:Clinical data of 60 patients with CM Ⅰ and SM treated in the Department of Neurosurgery of Peking University Third Hospital from March 2013 to November 2022 were analyzed retrospectively.There were 18 males and 42 females with an average age of 48.8±9.1(range,27-65)years old.All the patients un-derwent PFD.Imaging assessments were performed preoperatively by cervical spine MRI and X-ray,and pa-rameters were collected,including maximum anterior-posterior and transverse diameters of syrinx,spinal cord anterior-posterior diameter,longitudinal distances of bulbo-pontine sulcus,fourth ventricle and cerebellar ton-sil,length of McRae line,length of clivus,clivus-canal angle,C0-C2 and C2-C7 Cobb angles as well as cervical sagittal vertical axis.The maximum syrinx/cord ratio and maximum syrinx area were calculated.With-in 1 month postoperatively,the maximum anterior-posterior diameter of syrinx was measured again by cervical spine MRI and its change rate was calculated to evaluate the early prognosis of syrinx.The patients were di-vided into improved(the change rate≥20％)and unimproved(the change rate＜20％)groups.The imaging find-ings were compared between the two groups,and a multi-variate logistic regression analysis was employed to identify significant predictors.Results:All the patients were followed up for 12.2±7.1d(range,5-31d).After PFD surgery,24 cases were improved in syrinx(the improved group),while 36 cases were not improved(the u-nimproved group).Age and gender were not significantly different between the two groups(P＞0.05).Postopera-tively,there were 2 cases with fever,2 cases with transient occipital pain,4 cases with CSF leakage,1 case with poor incision healing,and there was no significant difference of overall complication ratio between the two groups(P＞0.05).The clivus-canal angle and cervical sagittal vertical axis of CM Ⅰ patients in improved group(153.47°±9.99°,22.10±11.10mm)were significantly greater than those of unimproved patients(147.98°±9.55°,15.83±9.00mm)(P＜0.05).The maximum transverse diameter of syrinx in improved group(8.34±3.20mm)was significantly smaller than that of unimproved group(10.36±3.71mm,P＜0.05).No significant differences were found in other preoperative imaging parameters(P＞0.05).The multi-variate logisitic analysis revealed that the maximum transverse diameter of syrinx before operation was a risk factor of syrinx size reduction(OR=1.281,95％CI 1.042-1.576,P＜0.05),while the offset distance of cervical sagittal vertical axis acted as a protective factor of syrinx size reduction(OR=0.916,95％CI 0.853-0.984,P＜0.05).Conclusions:In patients with CM Ⅰand SM,preoperative maximum transverse diameter of syrinx and offset distance of cervical sagittal vertical axis are independent influencing factors for early postoperative prognosis of syrinx.

Objective To investigate the recovery of plasma metabolism in asymptomatic and mild patients of coronavirus disease 2019(COVID-19)one year after recovery.Methods A total of 174 participants were recruited from the communities in Wuhan,including 80 healthy volunteers and the COVID-19 patients who had recovered for one year.According to the disease severity,the recovered COVID-19 patients were grouped as asymptomatic patients(n=80)and mild patients(n=14).The liquid chromatography mass spectrometry platform was employed to study the metabolomic characteristics of the plasma from all the participants.Results The plasma metabolites in asymptomatic patients and mild patients remained abnormal compared with those in healthy volunteers.Among the differential metabolites in asymptomatic patients and mild patients,some metabolites showed a downward trend only in mild patients,such as phosphatidylethanolamine[20∶3(5Z,8Z,11Z)/P-18∶0],sphingomyelin(d18∶1/24∶0),and cholesteryl(15∶0).The metabolic pathway involving the differential metabolites in mild patients was mainly glycerophospholipid metabolism.Conclusions Even one year after recovery,the mild COVID-19 patients still exhibit metabolic abnormalities.Hence,these patients may experience an extended period of time for recovery.
Humans ; COVID-19/metabolism* ; Metabolomics ; SARS-CoV-2 ; Metabolome ; Female ; Male ; Adult ; Middle Aged

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

Objectives:To investigate the imaging factors influencing the early prognosis of syrinx in patients with Chiari malformation type Ⅰ(CM Ⅰ)and syringomyelia(SM)who underwent posterior fossa decompression(PFD).Methods:Clinical data of 60 patients with CM Ⅰ and SM treated in the Department of Neurosurgery of Peking University Third Hospital from March 2013 to November 2022 were analyzed retrospectively.There were 18 males and 42 females with an average age of 48.8±9.1(range,27-65)years old.All the patients un-derwent PFD.Imaging assessments were performed preoperatively by cervical spine MRI and X-ray,and pa-rameters were collected,including maximum anterior-posterior and transverse diameters of syrinx,spinal cord anterior-posterior diameter,longitudinal distances of bulbo-pontine sulcus,fourth ventricle and cerebellar ton-sil,length of McRae line,length of clivus,clivus-canal angle,C0-C2 and C2-C7 Cobb angles as well as cervical sagittal vertical axis.The maximum syrinx/cord ratio and maximum syrinx area were calculated.With-in 1 month postoperatively,the maximum anterior-posterior diameter of syrinx was measured again by cervical spine MRI and its change rate was calculated to evaluate the early prognosis of syrinx.The patients were di-vided into improved(the change rate≥20％)and unimproved(the change rate＜20％)groups.The imaging find-ings were compared between the two groups,and a multi-variate logistic regression analysis was employed to identify significant predictors.Results:All the patients were followed up for 12.2±7.1d(range,5-31d).After PFD surgery,24 cases were improved in syrinx(the improved group),while 36 cases were not improved(the u-nimproved group).Age and gender were not significantly different between the two groups(P＞0.05).Postopera-tively,there were 2 cases with fever,2 cases with transient occipital pain,4 cases with CSF leakage,1 case with poor incision healing,and there was no significant difference of overall complication ratio between the two groups(P＞0.05).The clivus-canal angle and cervical sagittal vertical axis of CM Ⅰ patients in improved group(153.47°±9.99°,22.10±11.10mm)were significantly greater than those of unimproved patients(147.98°±9.55°,15.83±9.00mm)(P＜0.05).The maximum transverse diameter of syrinx in improved group(8.34±3.20mm)was significantly smaller than that of unimproved group(10.36±3.71mm,P＜0.05).No significant differences were found in other preoperative imaging parameters(P＞0.05).The multi-variate logisitic analysis revealed that the maximum transverse diameter of syrinx before operation was a risk factor of syrinx size reduction(OR=1.281,95％CI 1.042-1.576,P＜0.05),while the offset distance of cervical sagittal vertical axis acted as a protective factor of syrinx size reduction(OR=0.916,95％CI 0.853-0.984,P＜0.05).Conclusions:In patients with CM Ⅰand SM,preoperative maximum transverse diameter of syrinx and offset distance of cervical sagittal vertical axis are independent influencing factors for early postoperative prognosis of syrinx.

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 investigate the clinical application value of wrist joint ulnar deviation supination 45° palmar oblique posi-tion in the diagnosis of scaphoid waist fracture and displacement.Methods The imaging and clinical data such as digital radiography(DR),CT of 93 wrist joint trauma patients were analyzed.The four position views including wrist joint anteroposterior+lateral view,scaphoid position,wrist joint ulnar deviation supination 45° palmar oblique position,scaphoid position+wrist joint ulnar devia-tion supination 45° palmar oblique position were analyzed by three readers.The consistency of the evaluation among different readers and the diagnostic efficiency of the diagnosis of scaphoid fracture and displacement were compared.Results The inter-observer agreement,sensitivity,specificity,accuracy,and other diagnostic efficiency of scaphoid waist fracture and displacement was evaluated,wrist joint ulnar deviation supination 45° palmar oblique position+scaphoid position and wrist joint ulnar deviation supination 45° palmar oblique position were better than those of scaphoid position and wrist joint anteroposterior+lateral view.The combination of wrist joint ulnar deviation supination 45° palmar oblique position+scaphoid position obtained the best diagnostic efficiency.Conclusion The wrist joint ulnar deviation supination 45° palmar oblique position shows the long axis of the scaphoid,which has a high diagnostic efficiency in the diagnosis of scaphoid waist fracture and displacement and would be used as a useful supplement to other scaphoid imaging.

Objective To establish a dynamic prediction model of fatal massive hemorrhage in trauma based on the vital signs time series data and machine learning algorithms.Methods Retrospectively analyze the vital signs time series data of 7522 patients with trauma in the Medical Information Mart for Intensive Care-Ⅳ(MIMIC-Ⅳ)database from 2008 to 2019.According to the occurrence of posttraumatic fatal massive hemorrhage,the patients were divided into two groups:fatal massive hemorrhage group(n=283)and non-fatal massive hemorrhage group(n=7239).Six machine learning algorithms,including logistic regression(LR),support vector machine(SVM),random forests(RF),adaptive boosting(AdaBoost),gated recurrent unit(GRU),and GRU-D were used to develop a dynamic prediction models of fatal massive hemorrhage in trauma.The probability of fatal massive hemorrhage in the following 1,2,and 3 h was dynamically predicted.The performance of the models was evaluated by accuracy,sensitivity,specificity,positive predictive value,negative predictive value,Youden index,and area under receiver operating characteristic curve(AUC).The models were externally validated based on the trauma database of the Chinese PLA General Hospital.Results In the MIMIC-Ⅳ database,the set of dynamic prediction models based on the GRU-D algorithm was the best.The AUC for predicting fatal major bleeding in the next 1,2,and 3 h were 0.946±0.029,0.940±0.032,and 0.943±0.034,respectively,and there was no significant difference(P=0.905).In the trauma dataset,GRU-D model achieved the best external validation effect.The AUC for predicting fatal major bleeding in the next 1,2,and 3 h were 0.779±0.013,0.780±0.008,and 0.778±0.009,respectively,and there was no significant difference(P=0.181).This set of models was deployed in a public web calculator and hospital emergency department information system,which is convenient for the public and medical staff to use and validate the model.Conclusion A set of dynamic prediction models has been successfully developed and validated,which is greatly significant for the early diagnosis and dynamic prediction of fatal massive hemorrhage in trauma.

Objective To investigate the effects of leptin on hypothalamic neuron activity and lipid metabolism in adipose tissue of obese mice.Methods 10 leptin-deficient obese(ob/ob)mice with homozygous mutation of leptin gene and 10 wild-type(WT)mice born in the same litter were randomly divided into control group and leptin treatment group.The activity of pro-opiomelanocortin(POMC)neurons and tyrosine hydroxylase(TH+)neurons,the morphological changes of adipose tissue and the expression of lipid-related genes were analyzed by immunofluorescent staining,HE staining and Real-time PCR.Results Compared with the WT mice,the ob/ob mice showed decreased activity of POMC neurons and TH+neurons and larger cell diameter in adipose tissue and liver tissue.In addition,the expressions of heat-related genes uncoupling protein 1(UCP1),cytochrome c oxidase subunit 8B(Cox8b)and cell death-inducing DNA fragmentation factor,alpha subunit-like effector A(Cidea)in subcutaneous white fat in ob/ob mice decreased significantly,and the expressions of lipid synthesis-related genes sterol regulatory element binding transcription factor 1(Srebp1)and fatty acid synthase(Fas)increased significantly.After treated with leptin,the activities of POMC and TH+neurons were increased,and the cellular diameter and the degree of vacuolar degeneration were reduced in the adipose tissue and liver.Further analyses showed that the expressions of thermogenesis-related genes and lipolysis-related genes were increased,but expressions of lipid synthesis-related genes were reduced in brown adipose tissue.Conclusion Leptin treatment could prevent the increasing of obesity in ob/ob mice,which is associated with increased lipolysis and reduced lipid synthesis through activation of hypothalamic POMC neurons and peripheral adipose tissue sympathetic nervous system.

Objective To observe the clinical efficacy and safety of spironolactone combined with sacubitril/valsartan in the treatment of patients with hypertensive nephropathy.Methods The patients with hypertensive nephropathy were randomly divided into control group and treatment group.The control group was treated with sacubitril/valsartan(100-200 mg·d-1 in the morning),and treatment group was combined with low-dose spironolactone treatment(20 mg·d-1 in the morning)on the basis of control group.Both groups were treated continuously for 12 weeks.The clinical efficacy was compared;the blood pressure,urinary microalbumin(mAlb),urinary β2 microglobulin(β2-MG)and serum cystatin C(Cys-C),transforming growth factor-β1(TGF-β1),connective tissue growth factor(CTGF)and angiotensin Ⅱ(Ang Ⅱ)and adverse drug reactions were observed before and after treatment.Results There were 87 cases in treatment group and 86 cases in control group were included respectively.After treatment,the total effective rates in treatment group and control group were 95.40％(83 cases/87 cases)and 82.56％(71 cases/86 cases),with significant difference(P＜0.05).After treatment,the systolic blood pressure values in treatment group and control group were(124.65±9.65)and(130.27±8.93)mmHg,the diastolic blood pressure values were(75.08±7.14)and(80.45±7.35)mmHg,urinary mAlb levels were(42.58±5.65)and(51.28±6.64)mg·L-1,urinary β2-MG levels were(0.46±0.17)and(0.75±0.25)mg·L-1,24 h urinary protein quantitation levels were(138.49±46.64)and(216.48±65.27)mg,serum Cys-C levels were(0.63±0.26)and(0.85±0.24)mg·L-1,TGF-β1 levels were(98.67±21.43)and(112.46±26.72)pg·mL-1,CTGF levels were(1 206.54±236.56)and(1 340.51±248.25)pg·mL-1,Ang Ⅱ levels were(101.55±17.62)and(115.65±20.08)pg·mL-1,all with significant difference(all P＜0.05).The incidence of adverse drug reactions in treatment group and control group were 6.90％(6 cases/87 cases)and 2.33％(2 cases/86 cases),with no significant difference(P＞0.05).Conclusion Compared with sacubitril/valsartan alone,spironolactone combined with sacubitril/valsartan can better reduce blood pressure,improve renal function and delay progression of renal fibrosis in the treatment of hypertensive nephropathy,and has definite efficacy,with good safety.