Objective:To investigate the clinical significance of elevated cytokeratin 19 fragment (CYFRA21-1) in patients with dermatomyositis associated with positive anti-melanoma differentiation-associated gene 5 (MDA5) antibody.Methods:142 consecutive cases with newly onset anti-MDA5(+) (MADEDM)-DM admitted to the first affiliated hospital of Zhengzhou University from June 2018 to October 2021 were enrolled. They were divided into two groups, the low serum CYFRA21-1 group (CYFRA21-1≤4 ng/ml) and the high serum CYFRA21-1 group (CYFRA21-1>4 ng/ml). The clinical manifestations, laboratory tests results, imaging examinations treatment and outcome were collected for statistical analysis. Enumeration data were expressed as the number of cases and percentage (%). Normally distributed parameters were tested by t-test. Parameters with skewed distribution were tested by Mann-Whitney Wilcoxon analysis. Categorical variables were compared by the Chi-square test or Fisher′s exact test. Risk factor analysis was performed using Logistic regression. Cumulative survivals were described by Kaplan-Meier curves. Results:The age of onset in the high CYFRA21-1 group [(56±9)years vs. (50±10) years, t=-3.50, P=0.001] was higher than that in the low CYFRA21-1 group. Fever [63.3% (38/60) vs. 40.2% (33/82), χ2=7.39, P=0.007] was more common in the high CYFRA21-1 group, and arthritis [41.7% (25/60) vs. 69.5%(57/82), χ2=11.01, P=0.001] was less common. Myalgia, myasthenia, rashes, Raynaud′s phenomenon and skin ulcers had no significant difference between the two groups. The WBC count [5.2(4.1, 6.9)×10 9/L vs. 4.3(3.2, 6.2)×10 9/L, Z=-2.57, P=0.010], neutrophil count [4.0(2.9, 5.5)×10 9/L vs. 2.9(2.1, 4.5)×10 9/L, Z=-3.25, P=0.001] and neutrophil/lymphocyte ratio [5.75(3.50, 9.20) vs. 3.55(2.64, 5.41), Z=-3.77, P<0.001] in high CYFRA21-1 group were significantly higher than those in low CYFRA21-1 group. At the same time, LDH [384(302, 519)U/L vs. 318(260, 405)U/L, Z=-2.98, P=0.003], ferritin [1 204(677, 2 039)ng/ml vs. 570(229, 846)ng/ml, Z=-4.78, P<0.001], KL-6 [995(658, 1 491)U/ml vs. 750(563, 1 197)U/ml, Z=-2.49, P=0.013], ESR [36(22, 61)mm/1 h vs. 28(15, 46)mm/1 h, Z=-2.18, P=0.029] and CRP [9.2(4.7, 31.5)mg/L vs. 3.1(1.1, 11.6)mg/L, Z=-3.53, P<0.001] were significantly increased in the high level of CYFRA21-1 group, while serum albumin[(32±5)g/L vs. (35±5)g/L, t=3.92, P<0.001] was significantly decreased. There was no significant difference in the titers of serum anti-MDA5 antibodies between the two groups. The positive rate of anti-RO52 antibody [44(74.6%) vs. 44(53.7%), χ2=6.40, P=0.011] in high CYFRA21-1 group was higher than that in low CYFRA21-1 group. The ground glass opacity (GGO) score [1.75(1.33, 2.42) vs. 1.09(0.67, 1.67), Z=-4.60, P<0.001] based on high resolution CT (HRCT) was also significantly increased in the CYFRA21-1 high level group. Compared with the low CYFRA21-1 group, the high CYFRA21-1 group had a higher probability of RP-ILD [48.3%(29/60) vs. 23.2%(19/82), χ2=9.80, P=0.002] and a higher 6-month mortality rate[48.3%(29/60) vs.13.4%(11/82), χ2=19.70, P<0.001]. Logistic regression analysis showed that age ≥53 years old [ OR(95% CI)=5.197(1.781, 15.165), P=0.003], duration < 2 months [ OR(95% CI)=3.314 (1.058, 10.378), P=0.040], NE/LYMP >5 [ OR(95% CI)=3.443(1.120, 10.586), P=0.031], CRP>5 mg/L[ OR(95% CI)=6.271(1.749, 22.480), P=0.005], CA125>14 U/ml[ OR(95% CI)=7.500 (2.409, 23.345), P=0.001] and CYFRA21-1>4 ng/ml[ OR(95% CI)=3.665(1.258, 10.676), P=0.017] were independent risk factors for death within 6 months in MDA5-DM patients. Kaplan-Meier survival curve showed that the survival rate of the high CYFRA21-1 group was significantly lower than that of the low CYFRA21-1 group( P<0.001). Conclusion:Elevated CYFRA21-1 is an independent risk factor for early mortality in MDA5-DM patients and can serve as a novel serological marker for risk stratification in these patients.

Objective To investigate the relationship between serum trimethylamine oxide(TMAO),endo-thelial cell specific molecule 1(Endocan)and cardiac function and pregnancy outcome in patients with hyper-tensive disorders of pregnancy(HDP).Methods A total of 182 patients with HDP admitted to Handan Ma-ternal and Child Health Hospital from January 2021 to June 2023(HDP group)and 98 healthy pregnant women admitted to this hospital during the same period(control group)were selected as research subjects.Serum TMAO,Endocan and left ventricular cardiac function indexes[left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV)and left ventricular end-systolic volume(LVESV)]were compared between the two groups.According to pregnancy outcome,HDP patients were divided into poor outcome group(78 cases)and good outcome group(104 cases).Spearman correlation analysis was used to analyze the correlation between serum TMAO and Endocan and cardiac function indexes in HDP patients,and multi-factor Logistic regression was used to analyze the influencing factors of adverse pregnancy outcomes in HDP patients.The predictive value of serum TMAO and Endocan for adverse pregnancy outcomes in HDP patients was analyzed by receiver operating characteristic(ROC)curve.Results Compared with control group,serum TMAO,Endocan,LVEDV and LVESV were increased in HDP group,and LVEF was decreased(P＜0.05).Serum TMAO and Endocan in HDP patients were negatively correlated with LVEF(P＜0.05),and positively correlated with LVEDV and LVESV(P＜0.05).The incidence of adverse pregnancy outcomes in 182 HDP patients was 42.86％(78/182).Preeclampsia(PE),severe preeclampsia(SPE),24 h urine protein increase,LVEDV increase,LVESV increase,TMAO increase,Endocan increase were independent risk factors for adverse pregnancy outcomes in HDP patients,and LVEF increase was protective factor(P＜0.05).The area under the curve of serum TMAO combined with Endocan in predicting adverse pregnancy outcomes in HDP patients was 0.880,which was greater than 0.793 and 0.788 predicted by serum TMAO and Endocan a-lone.Conclusion The increase of serum TMAO and Endocan levels in HDP patients are relate to the decrease of cardiac function and adverse pregnancy outcomes,and the combined detection of the two has high predictive value for adverse pregnancy outcomes in HDP patients.

Objective To explore the relationship between serum pregnancy-specific protein 1(PSG1),stress-induced protein 2(Sestrin 2),growth arrest-specific protein 6(Gas6)and uterine artery blood flow pa-rameters and fetal growth restriction(FGR)in patients with gestational hypertension(GH).Methods A to-tal of 485 GH patients admitted to Handan Maternal and Child Health Hospital from January 2020 to October 2023 were selected as the research objects and divided into the occurrence group(81 cases)and the non-occur-rence group according to whether FGR occurred.The correlations between serum PSG1,Sestrin 2,Gas6 and uterine artery blood flow parameters[pulse index(PI),resistance index(RI),ratio of peak systolic velocity to end diastolic velocity(S/D)]was analysed,as well as the related influencing factors of FGR in GH patients.In addition,a Nomogram model based on the influencing factors was constructed to analyze the predictive value.Results The serum PSG1 level in occurrence group was significantly lower than that in non-occurrence group,and the serum Sestrin 2,Gas6 levels and PI,RI,S/D values were significantly higher than those in non-occurrence group(P＜0.05).Pearson correlation results showed that serum PSG1 was negatively correlated with the uterine artery blood flow parameters PI,RI,and S/D,and the levels of serum Sestrin 2 and Gas6 were positively correlated with the uterine artery blood flow parameters PI,RI,and S/D(P＜0.05).Gestational di-abetes mellitus,umbilical cord abnormalities,high Sestrin 2,high Gas6,high PI,high RI,and high S/D were independent risk factors for the occurrence of FGR in GH patients(P＜0.05),and increased PSG1 level was protective factor for the occurrence of FGR in GH patients(P＜0.05).Receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)of the Nomogram prediction model con-structed based on the influencing factors for predicting the occurrence of FGR in GH patients was 0.982,and the sensitivity and specificity were 0.943 and 0.938,respectively.The internal verification of the Bootstrap method shows that the Bias-corrected prediction curve basically coincides with the Ideal line,and the consis-tency index(C-index)was 0.964,indicating that the model was relatively stable.The decision curve shows that the threshold probability of this model was 0.01-1.00 and the net return rate was above 0.Conclusion Ser-um PSG1,Sestrin 2 and Gas6 in GH patients are closely related to uterine artery blood flow parameters and FGR,and the three are the influencing factors for the occurrence of FGR in GH patients.The constructed No-mogram model has a good predictive efficacy for FGR.

Objective To analyze the effects of tyrosinase on choroidal and retinal thickness and blood flow changes in guinea pigs,and to explore the role of tyrosinase in the development and progression of myopia.Methods A total of 30 three-week-old male tri-colored guinea pigs and 10 albino guinea pigs were selected and divided into four groups:control group(tri-colored hyperopic guinea pigs,with no intervention),albino group(albino myopic guinea pigs,with no inter-vention),myopia group(tri-colored myopic guinea pigs,with no intervention),and injection group[tri-colored hyperopic guinea pigs,injected with tyrosinase inhibitor(6 250 μg·L-1),100 μL per day].The experiment lasted for 4 weeks.The refractive status and axial length(AL)of the guinea pigs in each group were measured,along with ocular biometric param-eters related to axial length[anterior chamber depth,aqueous humor depth(AQD),central corneal thickness,lens diame-ter,vitreous chamber depth(VCD)].Optical coherence tomography was used to measure the retinal thickness,choroidal thickness(ChT),and choroidal blood perfusion(ChBP)in different regions of the guinea pigs.The tyrosinase activity in the vitreous and retina of guinea pigs in each group was measured.The expression levels of neurotransmitters in the vitre-ous and retina of guinea pigs in each group were detected.Results The differences in refractive status between the albi-no group and the control group at 0,2,and 4 weeks of the experiment were statistically significant(F=8.972,P＜0.05).At the 4th week of the experiment,the refractive status of the injection group was lower than that of the control group,and the AL was greater than that of the control group,with statistically significant differences(both P＜0.05).Further analysis of AL-related biometric parameters revealed that only AQD and VCD were associated with the changes in AL of guinea pigs in each group.At the 0th week of the experiment,the average retinal thickness of the control group was greater than that of the albino group,with a statistically significant difference(t=-9.007,P＜0.000 1).Moreover,the differences in reti-nal thickness in the outer retina across different directions and time points were statistically significant(all P＜0.05).The differences in retinal thickness between the control and albino groups were mainly concentrated in the outer retina.The ChT of the albino group was less than that of the control group,with a significant difference between groups(F=4.809,P=0.030).The ChBP of the control group was significantly higher than that of the albino group at 0,2,and 4 weeks of the experiment(all P＜0.05).At the 4th week of the experiment,the tyrosinase activity in the vitreous of the injection,albi-no,and myopia groups was lower than that of the control group,with statistically significant differences(all P＜0.05).The differences in neurotransmitters between the albino and control groups were mainly concentrated in the vitreous.In the retina,the ornithine level in the myopia group was higher than that in the albino and control groups,and the tryptophan levels in the myopia and control groups were higher than that in the albino group,with statistically significant differences(all P＜0.05).Conclusion Tyrosinase plays a crucial role in the development of myopia,regulating the development of refractive status by influencing the physiological properties of the retina and choroid.

Objective To analyze the factors influencing the deterioration of nutritional status after radiotherapy/radiochemotherapy for esophageal cancer,so as to provide reference for nutritional management during antitumor therapy.Methods A total of 106 patients with esophageal cancer who received radiotherapy or radiochemotherapy at Anqing First People's Hospital of Anhui Medical University from Dec.2017 to Dec.2023 were enrolled.Patients'gender,age,surgical history,timing of radiotherapy intervention,synchronous chemoradiotherapy,radiotherapy dose,clinical stage,initial nutritional status,and performance status score were collected.The patient generated subjective global assessment scale(PG-SGA)scores were monitored before and after antitumor treatment.According to the nutritional status at the beginning of enrollment and at the end of radiotherapy,the patients were assigned to deterioration group or non-deterioration(stable or improved)group.The clinical characteristics of the 2 groups were compared.The factors influencing the deterioration of nutritional status were screened by logistic regression analysis.The correlation between nutritional status deterioration and adverse reactions(radiation esophagitis,pulmonary infection,neutropenia,thrombocytopenia,and elevated aminotransferase)was analyzed by Spearman correlation analysis.Results There were no significant differences in gender,radiotherapy dose,initial nutritional status,or performance status score between the 2 groups for the deterioration of nutritional status after radiotherapy(all P＞0.05).The proportions of patients with previous surgical history of esophageal cancer,synchronous chemoradiotherapy,initiation of radiotherapy at less than 90%of target calorie requirement,and clinical stage Ⅳ were significantly higher in the deterioration group than those in the non-deterioration group(all P＜0.05).Logistic regression analysis showed that clinical stage Ⅳ(odds ratio[OR]=4.684,95%confidence interval[CI]1.252-17.519,P=0.022)and previous surgical history of esophageal cancer(OR=7.338,95%CI 1.878-28.666,P=0.004)were the independent adverse risk factors for the deterioration of nutritional status after radiotherapy/radiochemotherapy.The timing of radiotherapy intervention was also an independent risk factor for the deterioration of nutritional status,and taking the tolerance of 70%-90%target energy as the reference level,starting radiotherapy when the tolerance of 90%-100%target energy had the optimal protection of nutritional status(OR=0.166,95%CI 0.050-0.551,P=0.003).Spearman correlation analysis showed that the deterioration of nutritional status was positively correlated with elevated transaminases after radiotherapy(rs=0.283,P=0.003),while it was not correlated with the other adverse reactions(all P＞0.05).Conclusion Under the standard nutritional intervention model,patients with previous surgery and recurrent metastatic esophageal cancer who receive radiotherapy/chemoradiotherapy are still at risk of nutritional status deterioration.Tolerance to 90%-100%target energy requirement may be a more appropriate timing for radiotherapy intervention.When the nutritional status deteriorates during treatment,it is necessary to be alert to the elevated transaminases.

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.

Objective:To compare the significantly differentially expressed proteins in peripheral blood mononuclear cells between patients with different types of depressive disorders and normal participants and explore objective biomarkers with diagnostic significance.Methods:From March to May 2024, peripheral blood samples were collected from bipolar depression (BP), simple depression (DEP), persistent depressive disorder (PDD), and normal participants (NP) at the outpatient department of the Mental Health Center of Ningxia Medical University General Hospital. Using data-independent acquisition (DIA) quantitative proteomics technology, differentially expressed proteins were identified in the peripheral blood of patients with different types of depressive disorders and normal participants, conducting bioinformatics analysis to obtain candidate objective biomarkers, and finally identifying common and unique objective biomarkers for different types of depressive disorders. Data analysis was conducted using R software (version 4.0) and SPSS 27.0 software.Results:A total of 167 differentially expressed proteins were screened between BP and NP, 30 differentially expressed proteins were screened between DEP and NP, and 14 differentially expressed proteins were screened between PDD and NP.The differential proteins unique to the BP group were ARPC4 SPARC、MTPN、SNCA、YWHAH、MIF、COTL1, the differentially expressed proteins unique to the DEP group were RBBP9 APOF、S100A6, and the unique differential proteins in the PDD group were APOA5 CA1、IGLV3-25、APOC2.Conclusion:The objective biomarkers and signaling pathways obtained using DIA quantitative proteomics technology are of great significance for revealing the biological basis and diagnosing accurately of different types of depressive disorders.

Objective:To compare the significantly differentially expressed proteins in peripheral blood mononuclear cells between patients with different types of depressive disorders and normal participants and explore objective biomarkers with diagnostic significance.Methods:From March to May 2024, peripheral blood samples were collected from bipolar depression (BP), simple depression (DEP), persistent depressive disorder (PDD), and normal participants (NP) at the outpatient department of the Mental Health Center of Ningxia Medical University General Hospital. Using data-independent acquisition (DIA) quantitative proteomics technology, differentially expressed proteins were identified in the peripheral blood of patients with different types of depressive disorders and normal participants, conducting bioinformatics analysis to obtain candidate objective biomarkers, and finally identifying common and unique objective biomarkers for different types of depressive disorders. Data analysis was conducted using R software (version 4.0) and SPSS 27.0 software.Results:A total of 167 differentially expressed proteins were screened between BP and NP, 30 differentially expressed proteins were screened between DEP and NP, and 14 differentially expressed proteins were screened between PDD and NP.The differential proteins unique to the BP group were ARPC4 SPARC、MTPN、SNCA、YWHAH、MIF、COTL1, the differentially expressed proteins unique to the DEP group were RBBP9 APOF、S100A6, and the unique differential proteins in the PDD group were APOA5 CA1、IGLV3-25、APOC2.Conclusion:The objective biomarkers and signaling pathways obtained using DIA quantitative proteomics technology are of great significance for revealing the biological basis and diagnosing accurately of different types of depressive disorders.

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.

Objective To analyze the effects of tyrosinase on choroidal and retinal thickness and blood flow changes in guinea pigs,and to explore the role of tyrosinase in the development and progression of myopia.Methods A total of 30 three-week-old male tri-colored guinea pigs and 10 albino guinea pigs were selected and divided into four groups:control group(tri-colored hyperopic guinea pigs,with no intervention),albino group(albino myopic guinea pigs,with no inter-vention),myopia group(tri-colored myopic guinea pigs,with no intervention),and injection group[tri-colored hyperopic guinea pigs,injected with tyrosinase inhibitor(6 250 μg·L-1),100 μL per day].The experiment lasted for 4 weeks.The refractive status and axial length(AL)of the guinea pigs in each group were measured,along with ocular biometric param-eters related to axial length[anterior chamber depth,aqueous humor depth(AQD),central corneal thickness,lens diame-ter,vitreous chamber depth(VCD)].Optical coherence tomography was used to measure the retinal thickness,choroidal thickness(ChT),and choroidal blood perfusion(ChBP)in different regions of the guinea pigs.The tyrosinase activity in the vitreous and retina of guinea pigs in each group was measured.The expression levels of neurotransmitters in the vitre-ous and retina of guinea pigs in each group were detected.Results The differences in refractive status between the albi-no group and the control group at 0,2,and 4 weeks of the experiment were statistically significant(F=8.972,P＜0.05).At the 4th week of the experiment,the refractive status of the injection group was lower than that of the control group,and the AL was greater than that of the control group,with statistically significant differences(both P＜0.05).Further analysis of AL-related biometric parameters revealed that only AQD and VCD were associated with the changes in AL of guinea pigs in each group.At the 0th week of the experiment,the average retinal thickness of the control group was greater than that of the albino group,with a statistically significant difference(t=-9.007,P＜0.000 1).Moreover,the differences in reti-nal thickness in the outer retina across different directions and time points were statistically significant(all P＜0.05).The differences in retinal thickness between the control and albino groups were mainly concentrated in the outer retina.The ChT of the albino group was less than that of the control group,with a significant difference between groups(F=4.809,P=0.030).The ChBP of the control group was significantly higher than that of the albino group at 0,2,and 4 weeks of the experiment(all P＜0.05).At the 4th week of the experiment,the tyrosinase activity in the vitreous of the injection,albi-no,and myopia groups was lower than that of the control group,with statistically significant differences(all P＜0.05).The differences in neurotransmitters between the albino and control groups were mainly concentrated in the vitreous.In the retina,the ornithine level in the myopia group was higher than that in the albino and control groups,and the tryptophan levels in the myopia and control groups were higher than that in the albino group,with statistically significant differences(all P＜0.05).Conclusion Tyrosinase plays a crucial role in the development of myopia,regulating the development of refractive status by influencing the physiological properties of the retina and choroid.