Objective:To explore the effect of combining repeated peripheral (rPMS) and central transcranial magnetic stimulation (rTMS) in treating upper limb motor dysfunction after a stroke.Methods:Seventy-eight patients with upper limb motor dysfunction after a stroke were randomly divided into a control group, an rTMS group and a combined magnetic stimulation group, each of 26. All three groups underwent routine rehabilitation, while the rTMS group was repeatedly given low frequency transcranial magnetic stimulation of the M1 region on the unaffected side, and the combined group also received repeated peripheral magnetic stimulation at Erb′s point on the affected upper limb. There was one treatment session a day, 5 days a week for 3 weeks. Before and after the treatment, everyone′s upper limb motor function was quantified using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Wolf motor function test (WMFT). Skill in the activities of daily living was quantified in terms of a Barthel index (BI). Motor recovery of the upper limbs and hands was assessed using Brunnstrom staging. The latency and amplitude of the motor evoked potentials (MEPs) in the subjects′ affected abductor pollicis brevis muscles were also recorded before and after the treatment. Pearson correlation coefficients quantified the correlation between the changes in FMA-UE scores and MEP amplitudes before and after the treatment in the three groups.Results:There were no significant differences among the three groups before the treatment. Afterward, however, the average FMA-UE, WMFT and BI scores, as well as the upper limb and hand Brunnstrom stages and the average MEP latencies and amplitudes of all the three groups had improved significantly. The combined group′s average results were then significantly better than the other two groups′ averages, except for the upper limb Brunnstrom stages. The increases in MEP amplitude were positively correlated with the increases in FMA-UE scores among the rTMS and the combined group, but there was no significant correlation between them in the control group.Conclusions:The combined application of rPMS and contralateral low frequency rTMS can effectively relieve motor dysfunction in the upper limbs in the early stages after a stroke.

Objective To evaluate the intra-rater and inter-rater consistency of GRBAS and CAPE-V in Chi-nese context.Methods Five experts used the"Voice Assessment Assistant"APP to evaluate voice samples extrac-ted from The Sixth Medical Center of Chinese PLA General Hospital Pathological Voice Database V1.0.Fifty-two cases were used for inter-rater consistency analysis,and 38 cases were used for intra-rater consistency assessment.Intraclass correlation coefficient(ICC)was used for intra-rater and inter-rater consistency analysis of CAPE-V,while Cohen's Kappa method was applied for intra-rater consistency analysis of the five features of GRBAS.Inter-rater consistency analysis for GRBAS used Fleiss Kappa method,and Spearman's correlation analysis was conducted for the same features in both scales.Results The results of intra-rater consistency analysis showed that all six fea-tures of CAPE-V exhibited high consistency,with ICC coefficients of overall severity(OS)=0.80,roughness(R)=0.69,breathiness(B)=0.77,strain(S)=0.75,pitch(P)=0.74,and loudness(L)=0.78.For GRBAS,ex-cept for the features G(0.48)and S(0.45)which showed weaker intra-rater consistency,the rest of the features had poor intra-rater consistency.The results of inter-rater consistency analysis showed that the ICC correlation coef-ficients for all features of CAPE-V were greater than 0.85,indicating a high degree of consistency in the scoring of each feature by different raters.For GRBAS,except for the feature G(correlation coefficient=0.48),the rest of the features had correlation coefficients less than 0.40,indicating that the inter-rater consistency was poor for all features except G.When evaluating the same sample with both GRBAS and CAPE-V,the Spearman correlation co-efficients for the overall voice quality OS/G,roughness R,breathiness B,and strain S were 0.89,0.85,0.91,and 0.91,respectively,suggesting a high degree of correlation between the scoring results of the four features in the two scales.Conclusion In the auditory perceptual assessment of Chinese voice samples,the CAPE-V scale has higher intra-rater and inter-rater consistency compared to the GRBAS scale and is more suitable for clinical practice.

Objective To evaluate the intra-rater and inter-rater consistency of GRBAS and CAPE-V in Chi-nese context.Methods Five experts used the"Voice Assessment Assistant"APP to evaluate voice samples extrac-ted from The Sixth Medical Center of Chinese PLA General Hospital Pathological Voice Database V1.0.Fifty-two cases were used for inter-rater consistency analysis,and 38 cases were used for intra-rater consistency assessment.Intraclass correlation coefficient(ICC)was used for intra-rater and inter-rater consistency analysis of CAPE-V,while Cohen's Kappa method was applied for intra-rater consistency analysis of the five features of GRBAS.Inter-rater consistency analysis for GRBAS used Fleiss Kappa method,and Spearman's correlation analysis was conducted for the same features in both scales.Results The results of intra-rater consistency analysis showed that all six fea-tures of CAPE-V exhibited high consistency,with ICC coefficients of overall severity(OS)=0.80,roughness(R)=0.69,breathiness(B)=0.77,strain(S)=0.75,pitch(P)=0.74,and loudness(L)=0.78.For GRBAS,ex-cept for the features G(0.48)and S(0.45)which showed weaker intra-rater consistency,the rest of the features had poor intra-rater consistency.The results of inter-rater consistency analysis showed that the ICC correlation coef-ficients for all features of CAPE-V were greater than 0.85,indicating a high degree of consistency in the scoring of each feature by different raters.For GRBAS,except for the feature G(correlation coefficient=0.48),the rest of the features had correlation coefficients less than 0.40,indicating that the inter-rater consistency was poor for all features except G.When evaluating the same sample with both GRBAS and CAPE-V,the Spearman correlation co-efficients for the overall voice quality OS/G,roughness R,breathiness B,and strain S were 0.89,0.85,0.91,and 0.91,respectively,suggesting a high degree of correlation between the scoring results of the four features in the two scales.Conclusion In the auditory perceptual assessment of Chinese voice samples,the CAPE-V scale has higher intra-rater and inter-rater consistency compared to the GRBAS scale and is more suitable for clinical practice.

Objective:To explore the effect of combining repeated peripheral (rPMS) and central transcranial magnetic stimulation (rTMS) in treating upper limb motor dysfunction after a stroke.Methods:Seventy-eight patients with upper limb motor dysfunction after a stroke were randomly divided into a control group, an rTMS group and a combined magnetic stimulation group, each of 26. All three groups underwent routine rehabilitation, while the rTMS group was repeatedly given low frequency transcranial magnetic stimulation of the M1 region on the unaffected side, and the combined group also received repeated peripheral magnetic stimulation at Erb′s point on the affected upper limb. There was one treatment session a day, 5 days a week for 3 weeks. Before and after the treatment, everyone′s upper limb motor function was quantified using the Fugl-Meyer upper extremity assessment (FMA-UE) and the Wolf motor function test (WMFT). Skill in the activities of daily living was quantified in terms of a Barthel index (BI). Motor recovery of the upper limbs and hands was assessed using Brunnstrom staging. The latency and amplitude of the motor evoked potentials (MEPs) in the subjects′ affected abductor pollicis brevis muscles were also recorded before and after the treatment. Pearson correlation coefficients quantified the correlation between the changes in FMA-UE scores and MEP amplitudes before and after the treatment in the three groups.Results:There were no significant differences among the three groups before the treatment. Afterward, however, the average FMA-UE, WMFT and BI scores, as well as the upper limb and hand Brunnstrom stages and the average MEP latencies and amplitudes of all the three groups had improved significantly. The combined group′s average results were then significantly better than the other two groups′ averages, except for the upper limb Brunnstrom stages. The increases in MEP amplitude were positively correlated with the increases in FMA-UE scores among the rTMS and the combined group, but there was no significant correlation between them in the control group.Conclusions:The combined application of rPMS and contralateral low frequency rTMS can effectively relieve motor dysfunction in the upper limbs in the early stages after a stroke.

Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.

Objective:Voice is a vital tool for human communication, and its health spans across various stages of an individual's life cycle. This article discuss the importance of voice health from a lifecycle perspective, exploring the needs of voice, phonatory organ changes, the main etiologies and their incidence, the impact on social function, and treatment of voice disorders at different stages （childhood, adolescence, adulthood, and old age）. During childhood, due to incomplete vocal cords development and weak behavioral control, the incidence of voice disorders is high, which adversely affects children's psychological and social function development. The most common condition is vocal nodules, and there has been insufficient attention to the treatment of voice disorders in this stage. Voice disorders during adolescence are mainly related to physiological development and hormonal changes. In the elderly, the larynx undergoes structural and functional aging, which can be delayed with active intervention and treatment. Therefor, maintaining voice health should focus on preventing and managing voice problems throughout the entire life cycle, with targeted voice care and intervention at each stage. Establishing a comprehensive voice health management framework thereby upholds an individual's optimal health status and social function.
Humans ; Voice Disorders/etiology* ; Adolescent ; Child ; Adult ; Voice/physiology* ; Voice Quality ; Vocal Cords ; Aging/physiology* ; Aged

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.

Objective:To evaluate the clinical efficacy of intermittent theta burst stimulation (iTBS) of the bilateral cerebellum in treating post-stroke dysphagia.Methods:Forty patients with dysphagia after a cerebral infarction were randomly divided into a control group and an iTBS group, with 20 cases in each group. In addition to routine swallowing rehabilitation training and nutritional guidance, the iTBS group underwent daily bilateral cerebellar iTBS 6 times a week for 3 weeks, while the control group was given sham stimulation. Before and after the treatment, both groups were evaluated by using the water-swallowing test and the standardized swallowing assessment (SSA), and the latency and amplitude of the mylohyoid muscle′s motor evoked potential (MEP) were also recorded. The SSA scores and MEP amplitudes of the mylohyoid muscle after treatment were tested for any correlation.Results:The treatment improved the water-swallowing test results in both groups, with that of the iTBS group significantly better than that of the control group. The clinical efficacy in the iTBS group was 95%, significantly higher than the control group′s 80%. The SSA scores decreased significantly in both groups after the treatment, with the iTBS group′s average score then significantly lower than that of the control group. The average latency and amplitude of the mylohyoid muscle′s MEP was also significantly better in the iTBS group after the treatment. In the control group only the average amplitude was significantly greater than before the treatment. Pearson correlation analysis showed that the SSA scores and the MEP amplitudes of the mylohyoid muscle after treatment were uncorrelated in both groups.Conclusions:Bilateral intermittent theta burst stimulation can effectively improve the swallowing of patients with post-stroke dysphagia.

Reproductive toxicology is a discipline that uses toxicological methods to study the mechanisms by which foreign substances interfere with the generation of eggs or sperm and their detrimental effects on offspring. Research includes evaluating the damaging effects of test substances on reproductive function of parents and the toxicity evaluation of offspring embryos. People are exposed to a wide range of drugs, chemicals and environmental pollutants on a daily basis, and determining whether these substances have reproductive toxicity is crucial for the health of future generations. Reproductive toxicology research is therefore critical. Given the specificity and importance of reproductive toxicity evaluation, corresponding institutions both domestically and internationally have issued guidelines, national standards, or industry standards, all of which involve animal experiments. In the study of reproductive system diseases, numerous animal models have been developed to investigate key reproductive organs, such as testicles and ovaries. Each model involves the selection of animals, the establishment of methods, and the quantification of evaluation indicators, and all have advantages and limitations. The choice of model should be based on experimental needs and the characteristics of the model. This paper summarizes commonly used animal models for reproductive and development toxicity evaluation in reproductive toxicology research, including rat models for fertility and early embryonic development toxicity, rat models for embryo-fetal development toxicity, rabbit models for embryo-fetal development toxicity evaluation, minipig models for embryo-fetal development toxicity, rat models for perinatal toxicity, zebrafish models for embryonic development toxicity, and models for evaluating ovarian toxicity induced by chemical drugs, radiotherapy, autoimmunity, and ovariectomy, as well as models for evaluating testicular toxicity caused by chemical drugs and environmental factors. The methods for establishing these models, their application scope, and characteristics are reviewed in order to provide references for relevant research applications.

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.