In order to obtain a recombinant fusion protein of porcine β-defensin-2 and interferon αwith broad-spectrum antibacterial and antiviral activities at the same time,the genes encoding both proteins were fused in series and inserted into the pPICZαA vector.This construct was then elec-trotransferred into Pichia pastoris KM71H cells to construct an engineered strain.Following methanol-induced expression,the recombinant protein was concentrated and isolated.The cytotox-icity of the protein was assessed using the MTT assay and a porcine red blood cell hemolysis test.Subsequently,the in vitro antibacterial and antiviral activities of the recombinant fusion protein were evaluated.The results showed that the engineered strain pPICZαA-PBD2-IFNα-KM71H of Pichia pastoris was successfully obtained,and the fusion protein PBD2-IFN-α was obtained by concentrating and purifying the fermentation broth after fermentation induction and expression.Its concentration was 1.116 g/L and molecular weight was 25 kDa.When the concentration of fusion protein was lower than 4-4 g/L,it had no obvious toxicity to PK-15 cells and porcine red blood cells.The diameter of the inhibition zone produced by the fusion protein on the mixed plate of Escherichia coli and Staphylococcus aureus was(15.0±0.9)mm,which had obvious antibacterial activity.The antiviral activity of the fusion protein against VSV in PK-15 cells was 8.89 × 105 U/mL measured by Reed-Muench method.This study laid a theoretical foundation for further develo-ping the recombinant fusion protein as an antibacterial and antiviral product.

In order to obtain a recombinant fusion protein of porcine β-defensin-2 and interferon αwith broad-spectrum antibacterial and antiviral activities at the same time,the genes encoding both proteins were fused in series and inserted into the pPICZαA vector.This construct was then elec-trotransferred into Pichia pastoris KM71H cells to construct an engineered strain.Following methanol-induced expression,the recombinant protein was concentrated and isolated.The cytotox-icity of the protein was assessed using the MTT assay and a porcine red blood cell hemolysis test.Subsequently,the in vitro antibacterial and antiviral activities of the recombinant fusion protein were evaluated.The results showed that the engineered strain pPICZαA-PBD2-IFNα-KM71H of Pichia pastoris was successfully obtained,and the fusion protein PBD2-IFN-α was obtained by concentrating and purifying the fermentation broth after fermentation induction and expression.Its concentration was 1.116 g/L and molecular weight was 25 kDa.When the concentration of fusion protein was lower than 4-4 g/L,it had no obvious toxicity to PK-15 cells and porcine red blood cells.The diameter of the inhibition zone produced by the fusion protein on the mixed plate of Escherichia coli and Staphylococcus aureus was(15.0±0.9)mm,which had obvious antibacterial activity.The antiviral activity of the fusion protein against VSV in PK-15 cells was 8.89 × 105 U/mL measured by Reed-Muench method.This study laid a theoretical foundation for further develo-ping the recombinant fusion protein as an antibacterial and antiviral product.

Objective     To analyze the clinical information of COVID-19 patients of Shanghai National Exhibition and Convention Center cabin hospital, and to explore the medical management strategy to provide thoughtful suggestions for other cabin hospitals and governments as valuable references. Methods     The clinical data of 174 308 patients confirmed COVID-19 in Shanghai National Exhibition and Convention Center cabin hospital from April 9 to May 31, 2022 were retrospectively reviewed. There were 103 539 male and 70 769 female patients, with an average age of 41.50±15.30 years. Medical and nursing management strategy was summarized. Results     Among the 174 308 patients, 71.5% (124 630 patients) were asymptomatic. The vaccination rate of patients with COVID-19 in the cabin hospital was 76.5% (133 338 patients), and the majority of none vaccinated patients were children under the age of 10 years and the elderly over the age of 60 years, the vaccination rate of whom was only 25.0% (1 322 patients) and 63.9% (13 715 patients), respectively. In addition, the proportion of mild symptom type in the patients not vaccinated was significantly higher than that in the vaccinated patients (P≤0.01). The average hospitalization time of patients in cabin hospital was 7.39±0.53 days, which was 7.01±2.12 days for patients under 60 years and 8.21±0.82 days for patients over 60 years. The hospitalization time of elderly patients was significantly longer (P≤0.01), and the hospitalization time of elderly patients at age over 60 years without vaccination was 8.94±1.71 days, which was significantly longer than the average hospitalization time and the time of elderly patients vaccinated (P≤0.01). The number of patients combined with basic diseases was 27 864 (16.0%), of which cardiovascular diseases accounted for 81.3% (22 653 patients). A total of 2 085 patients were transferred and treated in designated hospitals. Conclusion     Large scale cabin hospitals are helpful to cut off the source of infection. Attention shall be paid to the sorting of admission and timely transfer to other hospital during the patients management. Most of the patients have a good prognosis after treatment. The vaccination of key population and community-based screening will be the next step of focus.

Objective The aim of this study was to investi-gate the changes of brain function in patients with drug-naive idiopath-ic epilepsy ( DNIE ) using resting-state functional MRI ( rs-fMRI ) amplitude of low-frequency fluctuation ( ALFF) , analyze the correlation of abnormal brain regions with the clinical variable ( disease course) , and gain a deeper insight into the pathophysiological mechanisms of idiopathic epilepsy. Methods This study included 25 cases of DNIE (15 males and 10 females) and 34 cases of drug idiopathic epilepsy (DIE, 22 males and 12 females).Another 25 healthy volunteers matched with the DNIE patients in sex, age, education and handedness were recruited as normal controls.The rs-fMRI data obtained from all the subjects were processed, subjected to ALFF analysis, and compared among the DNIE, DIE, and nor-mal control groups.The correlation was evaluated between the ALFF statistical brain mapping and the course of disease. Results Obvious differences were found in ALFF among the DNIE, DIE and control subjects.Compared with the normal controls, the DNIE pa-tients showed increased ALFF in the right inferior temporal gyrus, right lingual gyrus and right cuneus, but decreased ALFF in the right insula, left hippocampus, right midbrain, right middle frontal gyrus, left anterior cingulated gyrus, left middle cingulate gyrus and right inferior parietal lobule.In comparison with the DIE patients, those of the DNIE group exhibited increased ALFF in the left inferior occipital gyrus, right middle occipital gyrus and left middle occipital gyrus, but decreased ALFF in the right inferior frontal gyrus, left insula, right superior temporal gyrus and right middle frontal gyrus.In the DNIE patients, the disease course was found to be correlated positively with ALFF in the right cerebellum posterior lobe, left cerebellar tonsil, right lingual gyrus, left orbital gyrus, left middle oc-cipital gyrus, left corpus callosum, left caudate nuclear, left superior frontal gyrus, left medial frontal gyrus, right precuneus and left middle frontal gyrus, but negatively with ALFF in the right parahippocampal, right superior temporal gyrus, left superior temporal gyrus and right post-central gyrus. Conclusion The ALFF of resting-state cerebral function is abnormal in DNIE patients.The correlation between ALFF and the clinical variable ( disease course) provides a new insight into the pathophysiological mechanisms of epilepsy.

Objective To develop a method for dynamically observing the biological efficacy of botulinum toxin A (BTX-A) and to investigate the dose-effect relationship between BTX-A dosage and muscle strength.MethodsFifty-four male Sprague Dawley rats were randomly divided into 9 groups.Groups 1-7 were injected intramuscularly with 0.1 ml BTX-A (0.01 U to 4.0 U) into the gastrocnemius on the right side.Rats in group 8 were injected intramuscularly with an equal volume of saline solution as the control group,and group 9 was used to determine the location of injection.Gastrocnemius muscle strength was evaluated using a self-made evaluation system before and after the toxin injection and on the 3rd,7th,14th,21st,30th,45th,60th and 75th day following.ResultsMuscle strength reached its lowest level on days 3 to 7,with a significant difference in the decline of muscle strength between the test groups and the control group up to day 60.With the lower BTX-A doses (0.01 U,0.1 U,0.5 U,1.0 U),muscle strength had decreased significantly on the 21st day,but recovered to its initial levels in all groups at the same time.There was no significant difference among the 1.0 U,1.5 U,2.0 U and 4.0 U groups.ConclusionsStandardized gastrocnemius injection combined with neuromuscular functional evaluation can establish a model of BTX-A dosage and muscle paralysis which can be used to assess the evolution of the biological efficacy of BTX-A.

Objective To find out the accurate resonance frequencies and vibration characteristics of intestine,lung,kidney,stomach,heart and liver of rat and rabbit in low frequency and to provide basic data for the study of organs' resonance injury and reaction.Methods Eight rats and eight rabbits were anaesthetized with 1.5％ sodium pentobarbital and acceleration sensor was fixed to the surface of their bowels.The rats were fastened on their back on the veneer hung horizontally by rubber band.The veneer was hammered upward from the down side,and the biggest value of frequency spectrum was considered the resonance frequency.Rabbits were fastened on their back on vibration platform and 2 mm amplitude vibration was given with the sweeping-frequency from 2 Hz to 50 Hz.The bowels amplitudes on each frequency point were measured.The frequency corresponding to the biggest amplitude was considered the resonance frequency.Results The resonance frequencies of rat intestine,lung,kidney,stomach,heart and liver were respectively 3.8,3.2,6.6,7.0,4,6 and 3.0 Hz within frequency interval of 0 Hz to 20 Hz.The resonance frequencies of rabbit intestine,lung,kidney,stomach,heart and liver were respectively 6.0,5.0,7.0,8.0,6.0 and 7.0 Hz within frequency interval of 2 Hz to 50 Hz.The vibration amplitude of lung was the biggest at the resonance frequency.The resonance frequencies of both rat and rabbit bowels showed no statistical correlations to body weights,bowel mass and bowel volumes.Conclusion Bowel resonance frequencies of rat and that of rabbit are evidently statistical different except for that of kidney.The resonance frequency values of rabbit bowels are closer to that of dog and monkey than to that of rat.Thus the resonance injury characteristic of rabbit bowels can be the research basis for dog,monkey,and even human bowels resonance injury.

Objective To study the characteristics of the sciatic nerve injuries inflicted by highvelocity triangle fragments in swine.Methods According to the injury position,14 swines weighing (34.29±5.2)kg were divided into two groups.All swines were shot by 0.37 g triangle fragments at (773.1±12.4)m/s aimed at the midpoint of the lateral body surface projection of the right sciatic nerve in Group A(n=7)and at 2 cm aside from sciatic nerve in Group B(n=7).The entrance,exit and length of wound tract,distance between nerve and wound tract were detected,and the pathology of sciatic nerve and wound tract at 48 hours after injury were observed under light microscope and electron microscope.Results The course of the fragments was deviated to different extent.Six sciatic nerves were in primary wound tract,four of which were lacerated and ruptured at different degrees.The light microscope showed pathological changes including severe hemorrhage and edema,leukocytic infiltration,neurotmesis,axonotmesis and light myelin coloration;while electron microscope showed severe degeneration of myelin sheath and neuraxon distortion.Six nerves(one in Group A and five in Group B)were in the concussion zone,with the distance between sciatic nerve and primary wound tract for(2.07±0.45)cm.Hyporrhea and hyperemia under perilemma and perineurium,partial nerve fiber disrupt and axonotmesis were observed by light microscope and partly myelin sheath delamination by electron microscope.Two nerves (one in Group A and another in Group B)were in the contusion zone,with moderate pathological changes.Conclusions The track of high velocity triangle fragments is unstable in the tissues.High disrupt rate,severe and wide extent lesion,early Wallerian degeneration are the main characteristics of direct injuries of swine's sciatic nerves.Indirect injuries can induce pathologic changes too,for which the lesion severity is related to the distance between nerves and primary wound tract.