To construct a safe and effective multi-epitope vaccine against the S1 protein of chicken infectious bronchitis virus(IBV).In this study,homologous and non-homologous dominant epitopes of IBV M41,T,QX and H120 virulent strain S1 proteins were screened by various online bioprediction software,respectively,and a new peptide W with high immunogenicity was construc-ted by connecting the screened B-cell and T-cell epitopes with a linker peptide.W was ligated to the truncated sequence of the four viral strains by T2A yietding to the eukaryotic expression vector pEGFP-N1,and it was identified by PCR and double digestion,the obtained recombinant plasmid was transfected into HEK293A cells and target protein expression was measured by Western blot.The constructed plasmid was injected intramuscularly twice to detect the antibody level,cytokine level,and peripheral blood T cell subsets were detected after two immunizations.The epitope pro-tein W was successfully constructed,which was structurally stable,antigenic,and soluble;the re-combinant plasmid pEGFP-WMQtH,pEGFP-W,and pEGFP-MQtH matched the expected size;anti-IBV IgG antibody levels in pEGFP-N1 was increased greatly compared to the PBS group.cyto-kines IL-2,and γ interferon(IFN-γ)were increased greatly(P＜0.05);peripheral blood CD4+/CD8a value(P＜0.05)was increased greatly.The W epitope protein was successfully constructed,which can effectively activate the humoral immunity and cellular immunity against four infectious bronchitis viruses(IBV),laying a foundation for the development of an effective vaccine against IB.

To construct a safe and effective multi-epitope vaccine against the S1 protein of chicken infectious bronchitis virus(IBV).In this study,homologous and non-homologous dominant epitopes of IBV M41,T,QX and H120 virulent strain S1 proteins were screened by various online bioprediction software,respectively,and a new peptide W with high immunogenicity was construc-ted by connecting the screened B-cell and T-cell epitopes with a linker peptide.W was ligated to the truncated sequence of the four viral strains by T2A yietding to the eukaryotic expression vector pEGFP-N1,and it was identified by PCR and double digestion,the obtained recombinant plasmid was transfected into HEK293A cells and target protein expression was measured by Western blot.The constructed plasmid was injected intramuscularly twice to detect the antibody level,cytokine level,and peripheral blood T cell subsets were detected after two immunizations.The epitope pro-tein W was successfully constructed,which was structurally stable,antigenic,and soluble;the re-combinant plasmid pEGFP-WMQtH,pEGFP-W,and pEGFP-MQtH matched the expected size;anti-IBV IgG antibody levels in pEGFP-N1 was increased greatly compared to the PBS group.cyto-kines IL-2,and γ interferon(IFN-γ)were increased greatly(P＜0.05);peripheral blood CD4+/CD8a value(P＜0.05)was increased greatly.The W epitope protein was successfully constructed,which can effectively activate the humoral immunity and cellular immunity against four infectious bronchitis viruses(IBV),laying a foundation for the development of an effective vaccine against IB.

Objective:To investigate the effects of thinned anterolateral thigh perforator flaps combined with finger splitting and webplasty in sequential treatment of degloving destructive wound of total hand.Methods:This study was a retrospective observational study. From January 2012 to January 2023, a total of 15 cases who met the inclusion criteria with degloving destructive wound of total hand were admitted to Ningbo No.6 Hospital, including 10 males and 5 females, aged 17-75 years. The wounds were all combined with exposed bones or tendon. Emergency debridement and vacuum sealing drainage were performed in all cases before flap transplantation in stage Ⅰ. After thorough debridement, the wound area was 11.0 cm×3.0 cm-23.0 cm×13.5 cm. One or both anterolateral thigh perforator flaps with size of 12.5 cm×5.0 cm-25.0 cm×15.5 cm were designed, cut, and thinned to repair the skin and soft tissue defects of the hand. The donor site was sutured directly or repaired with medium-thickness skin graft from the opposite thigh. As needed, the flap was reconstructed by finger splitting and webplasty once or more times every 3 months after stage Ⅰoperation. The survival and complications of flap and wound healing at the donor site were observed after stage Ⅰoperation. The appearance of flap, two-point discrimination distance, and hand function were observed during the follow-up. At the final follow-up, the function of the affected hand was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association.Results:After the operation of stage Ⅰ, all the flaps of 15 cases of patients survived completely, including 1 case that had arterial crisis of flap but survived completely after exploration and re-anastomosis of blood vessels; all the wounds at the donor site healed. During the follow-up period of 6 to 18 months after stage Ⅰ, the flap was slightly swollen, with a little pigmentation, and the two-point discrimination distance in the finger flap was 8-11 mm. The fingers could complete the basic life actions such as flexion, extension, pinch, and grip. At the final follow-up, 3 cases were excellent, 9 cases were good, and 3 cases were acceptable in function evaluation of the affected hand.Conclusions:For degloving destructive wound of total hand, free transplantation of one or both thinned anterolateral thigh perforator flaps is used for repair in stage Ⅰ, and finger splitting and webplasty are used to reconstruct the flaps in the later stage, which can basically restore the pinch and grip function of the affected hand that is required for daily life, and is worthy of clinical promotion.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

Objective:To investigate the clinical and electrophysiological characteristics of facial onset sensory motor neuronopathy (FOSMN) syndrome.Methods:Ten patients diagnosed with FOSMN syndrome in Peking Union Medical College Hospital from January 2012 to December 2022 were included. The clinical and electrophysiological characteristics of patients were analyzed and summarized, and the genetic testing was also performed in these patients.Results:The age of onset was (56.6±6.5) years, and the longest survival duration of disease was 10 years. All patients had numbness around the face and mouth as the first symptom and abnormal blink reflex. A total of 52 sensory nerve conduction nerves were detected, among which 2 median nerves and 2 μlnar nerves showed decreased amplitude of sensory nerve action potential. Needle electromyography showed neurogenic lesions, with both progressive and chronic denervation. Whole exome sequencing identified the heterozygous variant c.272A>C in the exon 4 of the SOD1 gene resulting in the amino acid change p.Asp90Ala in 1 patient. In all patients, the disease progressed relentlessly and eventually led to involvement of respiratory muscle. Conclusion:FOSMN syndrome is characterized by abnormal blink reflex and sometimes abnormal sensory nerve conduction may be shown on electrophysiologic testing.

Objective: To evaluate the value of cerebrospinal fluid markers expecially total-tau protein (T-tau), phosphorylated-tau protein (P-tau) in diagnosis and differentiation of sporadic Creutzfeldt-Jakob disease (sCJD). Methods: sCJD (according to 2009 Brain criteria, 2018 Neurology amended criteria), Alzheimer's disease (AD; the National Institute on Aging at National Institutes of Health and the Alzheimer's Association revised guidelines 2011 criteria) and other patients without cognitive impairment, matched for sex and age, in the Department of Neurology, Peking Union Medical College Hospital from 2018 to 2019 were enrolled. Twelve sCJD patients, 49 AD patients and 14 normal controls were enrolled. Cerebrospinal fluid (CSF) specimens were collected through gravity dropping directly, and further stored in -80 ℃ and disposed according to widely used standards. The levels of T-tau and P-tau were measured by ELISA. The data on electroencephalogram and neuroimaging findings of sCJD patients were recorded. Moreover, specimens of sCJD patients were sent to the Chinese Center for Disease Control and Prevention to test 14-3-3 protein and PRNP genotype. Results: Using Mann-Whitney U test, T-tau concentrations were found higher in patients with sCJD (1 211(448, 2 227) pg/ml) than in AD patients (549(314, 1 078) pg/ml; U=178, P=0.034 9), and both groups had higher T-tau than the control group (127(79, 192) pg/ml; U=20, 73, P<0.01). The level of P-tau was significantly increased in AD patients (72(58,109) pg/ml) compared to the control group (27(15, 42) pg/ml; U=82, P<0.01), but not in sCJD patients (32(24, 47) pg/ml). The T-tau/P-tau ratio was higher in sCJD patients (29.77(20.01, 54.53)) than in AD patients (7.45(4.79, 10.43); U=87, P<0.01). Twelve sCJD patients had cotical hyperintensity on diffusion weighted imaging and five had periodic three-phase waves on electroencephalogram. Nine sCJD patients, whose CSF samples were tested in the Chinese Center for Disease Control and Prevention, carried an M/M genotype at codon 129 and E/E at codon 219. Conclusion The CSF tau level and T-tau/P-tau ratio are significantly increased in sCJD, which may promote the diagnosis and differentiation of sCJD in routine clinical setting.

Immune checkpoint inhibitors (ICIs) can cause adverse reactions in the nervous system, with the incidence rate ranging from 0.1% to 12%, and 80% occurring within the first 4 months of ICI application. It can cause lesions in various parts of the nervous system, including aseptic meningitis, meningoencephalitis, necrotizing encephalitis, brainstem encephalitis, transverse myelitis and other central nervous system diseases. It can also cause cranial peripheral neuropathy, multifocal radicular neuropathy, Guillain-Barre syndrome, spinal radicular neuropathy and myasthenia gravis, myopathy, etc. For these complications of the nervous system, diagnosis could be made by sufficient collection of disease manifestations combined with imaging, cerebrospinal fluid examinations, electro-encephalogram or electro myography to exclude infection or tumor progression. In the treatment of severe cases, ICIs should be discontinued and treated with high doses of glucocorticoid or gamma globulin with systemic support. After neurological adverse reactions, the prognosis of severe cases is poor.

Objective: To summarize the clinical characteristics and nerve conduction damage in patients with early POEMS syndrome, and to explore the value of segment nerve conduction velocity in the diagnosis of POEMS syndrome. Methods: A total of 73 patients with POEMS syndrome and 27 healthy controls in Peking Union Medical College Hospital from September 2009 to June 2019 were recruited in this study. The motor and sensory nerve conduction characteristics of median, ulnar, tibial, and peroneal nerves and the clinical features of the participants were analyzed. The analysis parameters included: (1) distal motor latency (DML), compound muscle action potential (CMAP); (2) the median velocity from elbow to wrist, the median velocity from axillary to elbow, the ulnar velocity from the site below elbow to wrist, the ulnar velocity from the site above elbow to below elbow, the ulnar velocity from axillary to the site above elbow, the tibial velocity from ankle to knee, the peroneal velocity from ankle to fibulae capitulum; (3) sensory nerve conduction velocity and amplitude of these nerves; (4) terminal latency indices (TLI) of median; (5) motor nerve conduction blocks. Results: Peripheral nerve damages were the initial symptoms in thirty-two patients in this group, accounting for 43.8% (32/73), and 81.3% (26/32) of these patients only showed numbness in lower extremities. All POEMS syndrome patients with numbness had abnormal sensory nerve conduction, and 9.5% (7/73) of patients without sensory symptoms also had abnormal sensory nerve conduction. On the other hand, the decrease of CMAP amplitude corresponded to clinical muscle strength decline and motor dysfunction. In the patients with POEMS syndrome, motor nerve conduction in the lower limbs were more likely to be affected and the damages were more severe than in the upper limbs: the proportion of CMAP disappearance in the lower limbs and upper limbs was 47.6% (112/235) vs 2.8% (7/252; χ²=133.698, P<0.01). Sensory nerve conduction damage was more severe than motor nerve conduction: the proportion of amplitude disappearance in sensory and motor conduction was 43.0% (141/328) vs 24.4% (119/487; χ²=133.698, P<0.01). The slowing of motor nerve conduction velocity was more common than the decrease of CMAP amplitude: the rate of slowing down of motor nerve conduction was 88.7% (432/487), and the rate of decrease of amplitude was 52.8% (257/487; χ²=151.905, P<0.01). The DML of median and ulnar nerve in the POEMS syndrome group was longer than that in the control group (median nerve: 4.4 (3.7, 5.0) ms vs 3.2 (3.0, 3.5) ms, U=854.000, P<0.01; ulnar nerve: 3.1 (2.7, 3.8) ms vs 2.5 (2.2, 2.7) ms, U=1 077.500, P<0.01). The TLI of median nerve in patients with POEMS syndrome was significantly higher than that of healthy controls (0.40±0.11 vs 0.35±0.06, t=3.466, P=0.001). There was no statistically significant difference in nerve conduction velocity between the forearm segment and the upper arm segment in the POEMS group. Conclusions Neurological damages were common in patients with POEMS syndrome with sensory nerve damage often being the initial clinical manifestation. In patients with POEMS syndrome, the nerve damage in the lower limbs was more severe than in the upper limbs, and sensory nerve damage was more severe than motor nerve damage. Segmental motor nerve conduction results suggested that demyelination damage occurred earlier than axonal damage, and proximal lesions were slightly heavier than distal lesions. No conduction block was detected in these patients. These characteristics of segmental nerve conduction can provide more evidence for clinical diagnosis of POEMS syndrome.

Objective To characterize the brain microstructure changes of amyotrophic lateral sclerosis (ALS) patients with various levels of cognitive impairment as measured by diffusion tensor imaging (DTI).Methods A total of 55 ALS patients and 20 healthy controls (HC) were enrolled in the Department of Neurology of Peking Union Medical College Hospital From September 2013 to March 2017,and all participants underwent neuropsychological assessments and DTI scans.According to their cognitive performance,ALS patients were further subclassified into ALS with normal cognition (ALS-Cn,n =27),ALS with cognitive impairment (ALS-Ci,n =17) and ALS-frontotemporal dementia (ALS-FTD,n =11)subgroups.Comparisons of voxel-based and atlas-based fractional anisotropy (FA) and mean diffusivity (MD) data were conducted among the four subgroups.Results In the voxel-based analyses,the FA showed significant differences in cingulate gyms,corpus callosum,brain stem and cerebellum,and MD showed significant differences in bilateral frontal lobe,temporal lobe,cingulate gyms,corpus callosum,and cerebellum among the four subgroups.Besides,when compared to ALS-Ci,ALS-Cn and HC groups in the order,the areas of involvement were larger and differences were more significant in ALS-FTD group.In the atlas-based analyses,the FA and MD of the corticospinal tracts revealed no difference within the patients groups,but decreased FA and increased MD were found compared to HC group.The ALS-IFD patients manifested widespread white matter fiber integrity damage and microstructure impairment in the extramotor areas compared to other three groups.Conclusion The brain white matter structural patterns of ALS patients correlate with their cognitive function,and there is a gradient of alterations across the ALS-Cn,ALS-Ci and ALS-FTD continuum.