Encephalomyocarditis virus(EMCV)is a zoonotic pathogen that causes encephalitis and myocarditis as its primary clinical manifestations.To explore effective preventive measures,this study utilized a Bac-to-Bac expression system to insert the EMCV P12A and 3C genes into the pFastBacDual shuttle vector,resulting in the generation of the recombinant baculovirus Ac-P12A-3C.This facilitated the large-scale expression and purification of EMCV virus-like particles(VLPs),which were correctly assembled into particles of approximately 30 nm in diameter,as ob-served by electron microscopy.Immunization and challenge experiments in mice demonstrated that these VLPs could effectively protect against EMCV infection,achieving a protection rate of 100％.Histopathological sections indicated that,compared to the PBS control group,the VLP immuniza-tion group exhibited significantly reduced tissue damage,along with a marked decrease in viral load within the tissues.In piglets,immunization with the VLPs elicited a robust humoral response,with neutralizing antibody titers reaching 1∶320 to 1∶640 after a second immunization,and no signifi-cant adverse reactions were observed throughout the immunization process.This study preliminarily explores the immunogenicity and safety of the VLP vaccine,laying the foundation for the development of a subunit vaccine based on EMCV VLPs and offering a new strategy for the prevention and control of encephalomyocarditis.

At present, upper limb motor rehabilitation relies on specific rehabilitation aids, ignoring the initiative of upper limb motor of patients in the middle and late stages of rehabilitation. This paper proposes a fuzzy evaluation method for active participation based on trajectory error and surface electromyography (sEMG) for patients who gradually have the ability to generate active force. First, the level of motor participation was evaluated using trajectory error signals represented by computer vision. Then, the level of physiological participation was quantified based on muscle activation (MA) characterized by sEMG. Finally, the motor performance and physiological response parameters were input into the fuzzy inference system (FIS). This system was then used to construct the fuzzy decision tree (FDT), which ultimately outputs the active participation level. A controlled experiment of upper limb flexion and extension exercise in 16 healthy subjects demonstrated that the method presented in this paper was effective in quantifying difference in the active participation level of the upper limb in different force-generating states. The calculation results of this method and the active participation assessment method based on sEMG during the task cycle showed that the active participation evaluation values of both methods peaked in the initial cycle: (82.34 ± 9.3) % for this paper's method and (78.44 ± 7.31) % for the sEMG method. In the subsequent cycles, the values of both showed a dynamic change trend of rising first and then falling. Trend consistency verifies the effectiveness of the active participation assessment strategy in this paper, providing a new idea for quantifying the participation level of patients in middle and late stages of upper limb rehabilitation without special equipment mediation.
Humans ; Electromyography/methods* ; Upper Extremity/physiology* ; Fuzzy Logic ; Exercise Therapy/methods* ; Muscle, Skeletal/physiology* ; Male

Objective The effect of intra-aortic balloon counterpulsation combined with recombinant human brain natriuretic peptide(rhBNP)treatment on ventricular remodeling after percutaneous coronary intervention(PCI)in elderly patients with acute anterior wall myocardial infarction.Methods A retrospective analysis was con-ducted on 66 elderly patients with acute anterior myocardial infarction who underwent emergency PCI in our hospital from January 2018 to June 2023.They were divided into a control group(n=32)who underwent PCI+rhBNP,and a study group(n=32)who underwent PCI+rhBNP+intra-aortic balloon counterpulsation.Blood pressure,lactic acid level,mechanical complications,ventricular remodeling,inflammatory factors,myocardial enzymology,ox-LDL level were detected and compared between the two groups.Results Before treatment,Lactic acid,diastolic blood pressure,systolic blood pressure,left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),left ventricular posterior wall thickness(LVPWT),left ventricular remodeling index(LVRI),left ventricular ejection fraction(LVEF),homocysteine(Hcy),interleukin-6(IL-6)and myocardial troponin were compared between the two groups.Myocardial troponin T(cTnT),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),oxidized low density lipoprotein(ox-LDL)levels(P>0.05);At 3 months follow-up after discharge,the levels of lactic acid,LVESD,LVEDD,LVPWT,LVRI,Hcy,IL-6,cTnT,LDH,CK-MB and ox-LDL were decreased,and the levels were lower in the study group,and the levels of diastolic blood pressure,systolic blood pressure and LVEF were increased,and the levels were higher in the study group(P<0.05).The pro-portion of mechanical complications between the two groups before and after treatment were compared(P>0.05).Conclusion In elderly patients with acute anterior myocardial infarction after emergency PCI,intra-aortic balloon counterpulsation combined with rhBNP could alleviate ventricular remodeling,myocardial injury,reduce inflamma-tory injury,improve oxidative stress response,and regulate tissue perfusion.

Encephalomyocarditis virus(EMCV)is a zoonotic pathogen that causes encephalitis and myocarditis as its primary clinical manifestations.To explore effective preventive measures,this study utilized a Bac-to-Bac expression system to insert the EMCV P12A and 3C genes into the pFastBacDual shuttle vector,resulting in the generation of the recombinant baculovirus Ac-P12A-3C.This facilitated the large-scale expression and purification of EMCV virus-like particles(VLPs),which were correctly assembled into particles of approximately 30 nm in diameter,as ob-served by electron microscopy.Immunization and challenge experiments in mice demonstrated that these VLPs could effectively protect against EMCV infection,achieving a protection rate of 100％.Histopathological sections indicated that,compared to the PBS control group,the VLP immuniza-tion group exhibited significantly reduced tissue damage,along with a marked decrease in viral load within the tissues.In piglets,immunization with the VLPs elicited a robust humoral response,with neutralizing antibody titers reaching 1∶320 to 1∶640 after a second immunization,and no signifi-cant adverse reactions were observed throughout the immunization process.This study preliminarily explores the immunogenicity and safety of the VLP vaccine,laying the foundation for the development of a subunit vaccine based on EMCV VLPs and offering a new strategy for the prevention and control of encephalomyocarditis.

Objective The effect of intra-aortic balloon counterpulsation combined with recombinant human brain natriuretic peptide(rhBNP)treatment on ventricular remodeling after percutaneous coronary intervention(PCI)in elderly patients with acute anterior wall myocardial infarction.Methods A retrospective analysis was con-ducted on 66 elderly patients with acute anterior myocardial infarction who underwent emergency PCI in our hospital from January 2018 to June 2023.They were divided into a control group(n=32)who underwent PCI+rhBNP,and a study group(n=32)who underwent PCI+rhBNP+intra-aortic balloon counterpulsation.Blood pressure,lactic acid level,mechanical complications,ventricular remodeling,inflammatory factors,myocardial enzymology,ox-LDL level were detected and compared between the two groups.Results Before treatment,Lactic acid,diastolic blood pressure,systolic blood pressure,left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),left ventricular posterior wall thickness(LVPWT),left ventricular remodeling index(LVRI),left ventricular ejection fraction(LVEF),homocysteine(Hcy),interleukin-6(IL-6)and myocardial troponin were compared between the two groups.Myocardial troponin T(cTnT),lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),oxidized low density lipoprotein(ox-LDL)levels(P>0.05);At 3 months follow-up after discharge,the levels of lactic acid,LVESD,LVEDD,LVPWT,LVRI,Hcy,IL-6,cTnT,LDH,CK-MB and ox-LDL were decreased,and the levels were lower in the study group,and the levels of diastolic blood pressure,systolic blood pressure and LVEF were increased,and the levels were higher in the study group(P<0.05).The pro-portion of mechanical complications between the two groups before and after treatment were compared(P>0.05).Conclusion In elderly patients with acute anterior myocardial infarction after emergency PCI,intra-aortic balloon counterpulsation combined with rhBNP could alleviate ventricular remodeling,myocardial injury,reduce inflamma-tory injury,improve oxidative stress response,and regulate tissue perfusion.

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

A 27﹣year﹣old male patient underwent left maxillary masses excision for periapical cysts in the left maxillary under general anesthesia. During the surgery,a venous access was established from the peripheral vein of the patient′s left lower extremity,and anesthetic drugs(propofol injection,succinylcholine chloride injection,remifentanil hydrochloride for injection,dexmedetomidine hydrochloride injection)were given. On day 2 after surgery,the patient developed topical redness and swelling along the blood vessel on his left lower extremity,which gradually developed into a cord﹣like redness and swelling,with increased blood vessel stiffness. He was diagnosed with thrombophlebitis of the left great saphenous vein after a ultrasound examination of the blood vessel. Treatments such as subcutaneous injection of enoxaparin sodium, oral administration of extract of horse chestnut seeds, topical mucopolysaccharide polysulfate cream, physiotherapy,and etc. were given. On day 10 after surgery,the redness and swelling on the patient′s left lower extremity disappeared,and the blood vessel was softened. At 2 months of follow﹣up,ultrasound examination revealed partial recanalization of the left venous trunk( the calf part ) of saphenous vein. According to the drug labels and literature reports,it was considered that the patient′s thrombophlebitis might be associated with propofol injection.

A 12﹣year﹣old female pediatric patient received a continuous pumping of somatostatin 3 mg (added to 210 ml of 0. 9% sodium chloride injection)by an intravenous detaining needle on the back of her left hand and an IV infusion of pantoprazole sodium for injection 25 mg(added to 100 ml of 0. 9% sodium chloride injection)by an intravenous detaining needle on the back of her right hand at the same time for acute pancreatitis. About 5 minutes after the initiation of administration,cyanosis appeared on her both hands,her left hand was cold,and red rashes scattered on her left wrist and upper arm. It was considered as drug﹣induced anaphylaxis,and the drugs were discontinued immediately. About 40 minutes after drug withdrawal,her above symptoms relieved completely. Pantoprazole for injection was given again that day,and no allergic symptoms were observed. On day 2,a subcutaneous injection of 0. 1 g octreotide acetate injection( an analogue of somatostatin)was given. About 10 minutes later,the patient developed headache,nausea,and vomiting, which relieved spontaneously after 30 minutes. It was considered that the anaphylaxis was caused by somatostatin and its analogue octreotide.

A 27﹣year﹣old male patient underwent left maxillary masses excision for periapical cysts in the left maxillary under general anesthesia. During the surgery,a venous access was established from the peripheral vein of the patient′s left lower extremity,and anesthetic drugs(propofol injection,succinylcholine chloride injection,remifentanil hydrochloride for injection,dexmedetomidine hydrochloride injection)were given. On day 2 after surgery,the patient developed topical redness and swelling along the blood vessel on his left lower extremity,which gradually developed into a cord﹣like redness and swelling,with increased blood vessel stiffness. He was diagnosed with thrombophlebitis of the left great saphenous vein after a ultrasound examination of the blood vessel. Treatments such as subcutaneous injection of enoxaparin sodium, oral administration of extract of horse chestnut seeds, topical mucopolysaccharide polysulfate cream, physiotherapy,and etc. were given. On day 10 after surgery,the redness and swelling on the patient′s left lower extremity disappeared,and the blood vessel was softened. At 2 months of follow﹣up,ultrasound examination revealed partial recanalization of the left venous trunk( the calf part ) of saphenous vein. According to the drug labels and literature reports,it was considered that the patient′s thrombophlebitis might be associated with propofol injection.