Infectious bursal disease virus (IBDV) VP4 plays an important role in immunosuppression of host. In order to develop monoclonal antibodies (McAbs) against VP4, we vaccinated BALB/c mice with His-VP4, screened and subcloned positive clones. We established 4 hybridoma cell lines that stably secreted McAbs against VP4 and named these cell lines 3B3, 3H11, 4C8 and 4G6, respectively. We tested the dissociation constant (Kd) of these McAbs, and found that their K(d)s were 4.61 x 10(-11), 1.71 x 10(-10), 4.26 x 10(-11), 5.02 x 10(-11), respectively. The isotypes of these McAbs were determined to be IgG1, IgG1, IgG2b and IgG1. These McAbs specifically bound to VP4 in IBDV infected DF-1 cells as demonstrated by Western blotting analysis and fluorescence antibody assay. These McAbs would help to detect IBDV infection and to analyze the biological activities of IBDV VP4.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Blotting, Western ; Cell Line ; Fluorescent Antibody Technique ; Hybridomas ; Infectious bursal disease virus ; Mice ; Mice, Inbred BALB C ; Viral Structural Proteins ; immunology

Objective To estimate the safety and efficacy of submucosal tunneling endoscopic resection (STER) on treatment of large esophageal submucosal tumors (SMTs) originating from muscularis propria layer.Methods The data of patients with large esophageal SMTs (diameter ≥ 3.5 cm) undergone STER (n=17) or endoscopic submucosal dissection (ESD,n =15) at the Endoscopy Center of Tianjin Medical University General Hospital from December 2009 to March 2016 were retrospective analyzed.The therapeutic effects,hospitalization times,post-operation expenses,and occurrence of complications were evaluated and compared between the two groups.Results All the endoscopic treatments of the 32 patients were successfully completed.The operating time of the STER group was significantly longer than that of the ESD group (t =2.595,P =0.015).There was no statistical difference on the en bloc resection rate,complete resection rate and complication rate between STER group and ESD group (P＞0.05).The mean post-operative hospital stay of the STER group was significantly less than that of the ESD group (3.8± 1.0days VS 6.7±1.8 days,t=5.644,P=0.000).The mean hospital cost of the STER group was significantly less than that of the ESD group (22 456.1±5 232.0 yuan VS 27 392.5±5 747.9 yuan,t =2.543,P =0.016).The wound healing rates at 1 month after operation in the STER group was significantly higher than that of the ESD group [94.1％ (16/17) VS 20.0％ (3/15),P=0.000].No recurrence and metastasis occurred in the STER group and ESD group during the 41.2±20.6 months follow-up.Conclusion STER is a safe and effective technique for treating large esophageal SMTs originating from the muscularis propria layer,with earlier wound healing,shorter hospital stay and lower cost compared with those of the traditional method of ESD.

Immune checkpoint inhibitors (ICIs) targeting CTLA-4 and PD-1 /PD-L1 are increasingly used to treat several types of cancer.Although several agents have been approved,only 10％ to 30％ of patients have benefited from them.Some studies have shown that intestinal micro-ecology can affect the outcome of ICIs through immune regulation in cancer patients.This literature review s summarized the regulation and predictive functions of intestinal micro-ecology on ICIs treatment,and introduced possible mechanisms of intestinal micro-ecology that affacted the efficacy of immunotherapy.In addition,some approaches for detecting gut microbiome were also summarized.Gut microbiota highlighted in this review may serve as novel biomarkers to predict clinical outcomes in cancer patients.

Objective To observe the effect of ropivacaine on bilateral posterior thoracic paravertebral nerve block for cardiopulmonary bypass.Methods A total of 142 patients undergoing elective cardiac valve surgery,68 males and 74 females,were randomly divided into two groups:group C (general anesthesia,n =69) and group P (general anesthesia+ paraverteral nerve block with ropivacaine,n=73).The changes of hemodynamics,BIS,anesthesia dose and postoperative anesthesia were observed before and after anesthesia,the time of mechanical ventilation,the duration of vasoactive drug support,ICU stay time,length of stay and hospitalization costs were recorded.Results Compared with group C,heart rate and blood pressure were more stable in group P,and the dosage of sufentanil was less [(130.6±5.0) μg vs (202.8±7.4) μg,P＜0.01],the patient was awakened quickly [(241.0±15.8) min vs (417.6±38.9) min,P＜0.05],and the time of mechanical ventilation [(466.0±22.9) min vs (657.8±49.4) min,P＜0.05] and ICU residence time [(22.1±1.0) h vs (34.1±3.6) h,P＜0.05] were shortened,the cost of hospitalization decreased [(72 360±1 565) yuan vs (78 420±2 094) yuan,(P＜0.05)].Conclusion Thoracic paravertebral nerve block anesthesia combined with general anesthesia is better than pure general anesthesia,in terms of maintaining intraoperative circulation stability,reducing the amount of anesthetic analgesic drugs,and early recovery of patients after surgery.

Objective To observe the effect of Delta-like ligand 4 (Dll-4) on the pathological structure of retina in early diabetic rats (DM) and its relationship with vascular endothelial growth receptor-2 (VEGFR-2).Methods A total of 70 male Sprague-Dawley rats were randomly divided into normal group and DM group,with 10 and 60 rats in each group,respectively.The rats of DM group was induced by intraperitoneal injection of streptozotocin to established DM model.The rats with blood glucose recovery and death were excluded,and the final 60 rats were included in the statistics.Rats in the normal group were injected with an equal volume of citric acid-sodium citrate buffer.Rats in the DM group were divided into DM 1 month (DM lm) group,DM 2 months (DM 2m) group,DM 3 months (DM 3m) group and DM 3m + Anti group,DM 3m + phosphate buffer solution (PBS) group by random number table method,and 10 rats in each group.In the DM 3m+Anti group,4 μl ofantiDll-4 polyclonal antibody was injected into the vitreous cavity,and the antibody concentration was 0.25 mg/ml.The DM 3m+PBS group was intravitreally injected with an equal volume of PBS.Five days after the injection,the rats were sacrificed.Rats in the DM 3m group and the normal group were not treated,and were sacrificed 3 months after the model was established.The structure and microvascular changes of the retina were observed by hematoxylin-eosin staining,and the total thickness of the retina was measured.The expression of Dll-4 and VEGFR-2 in the retina was detected by immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR).One-way analysis of variance was used to compare the expression of Dll-4 and VEGFR-2 in the retina of each group.The least significant difference t test was used to compare the two groups.Results Light microscopy showed that the retinal ganglion cells layer in the DM 3m group were obviously edematous,the inner and outer nuclear layers were thinner,the number of cells was reduced,the arrangement was disordered,the edema of outer plexiform layer was obvious,and the microvessels were abnormally dilated.In the DM 3m+Anti group,the edema of outer plexiform layer was lessened than that of the DM 3m group,and the other layers were not significantly different from the DM 3m group.Compared with the normal group,the total retinal thickness of the DM 3m group,the DM 3m+Anti group and the DM 3m+PBS group increased (t=5.596,3.290,4.286;P=0.000,0.008,0.002).Immunohistochemical staining showed that a small amount of Dl14 was positively expressed in the retinal ganglion cell layer of the normal group;a small amount of VEGFR-2 was positively expressed in the ganglion cell layer and the inner and outer nuclear layers.The positive expression of Dll-4 and VEGFR-2 in retinal vascular endothelial cells of DM 3m group increased significantly.The expression of Dll-4 was significantly decreased in the retinal layers and vascular endothelial cells ofDM 3m+Anti group,while the expression of VEGFR-2 was significantly increased.There was no significant difference between the positive expression of Dll4 and VEGFR-2 in the DM 3m+PBS group and the DM 3m group.The results of real-time PCR showed that the relative expression of Dll-4 and VEGFR-2 mRNA in the DM 3m group was significantly higher than that in the normal group (t=6.705,20.871;P＜0.05).Compared with DM 3m group,the relative expression of Dll-4 mRNA in DM 3m+Anti group decreased,and the relative expression of VEGFR-2 mRNA increased (t=2.681,3.639;P＜0.05).The relative expressions of Dll-4 and VEGFR-2 mRNA in the DM 3m+PBS group and DM 3m group were not statistically significant (t=0.513,0.657;P＜0.05).Conclusions The expression of Dll-4 in retinal vascular endothelial cells is gradually increased during the early retinopathy of DM rats.The expression of Dll-4 is inhibited,the expression of VEGFR-2 is up-regulated,and the plexus edema is alleviated.

BackgroundElderly cataract patients with depressive symptoms have heavier self-perceived burden， and worse vision-related quality of life compared to patients without depressive symptoms， resulting in the increase of family burden. Most previous studies showed that family relationship and vision were the main factors leading to depression in elderly cataract patients， and ignored the effects of self-feeling and complicated diseases on the psychological state of elderly cataract patients. ObjectiveTo explore the relationship between depressive symptoms， self-perceived burden and postoperative vision-related quality of life in elderly cataract patients， and to analyze the risk factors of depressive symptoms. So as to provide references for targeted psychological intervention in this population. MethodsA total of 104 elderly cataract patients admitted to Jiangsu Province Hospital （The First Affiliated Hospital with Nanjing Medical University） from July 1， 2020 to December 31， 2022 were included. General information， self-perceived burden and postoperative vision-related quality of life were investigated by self-designed questionnaire， Patients' Health Questionnaire depressive Scale-9 item （PHQ-9）， Self-Perceived Burden Scale （SPBS） and National Eye Institute Visual Function Questionnaire-25 （NEI-VFQ-25）. Pearson correlation analysis was performed to analyze the correlation among PHQ-9， SPBS and NEI-VFQ-25 scores. Logistic regression analysis was used to analyze the influencing factors of depressive symptoms in eldery cataract patients. ResultsA total of 100 elderly cataract patients completed an effective questionnaire survey. Among the participants， 31 cases （31.00%） were found to have depressive symptoms. The depressive group exhibited significantly higher SPBS score （t=11.062， P<0.01） and significantly lower NEI-VFQ-25 score （t=-5.235， P<0.01） than those of the non-depressive group. Pearson correlation analysis showed a positive correlation between PHQ-9 and SPBS score （r=0.485， P<0.01）， and a negative correlation between PHQ-9 and NEI-VFQ-25 score （r=-0.440， P<0.01）. Complicated with diabetes （OR=1.441， P<0.01）， osteoarthritis （OR=1.324， P<0.05） and high SPBS score （OR=1.340， P<0.05） were risk factors of depressive sympptoms in elderly cataract patients. ConclusionThe detection rate of depressive symptoms in elderly cataract patients is higher， and postoperative vision-related quality of life in elderly cataract patients complicated with depressive symptoms is poor. Risk factors include diabetes， osteoarthritis and high self-perceived burden. ［Funded by 2022 Jiangsu Province Elderly Health Scientific Research Project （number， LKM2022019）］

Objective To explore the expression and their significance of peripheral Th17 cells and regulatory T cells (Tregs) in idiopathic inflammatory myopathy,and analyze the relationship between the expression and clinical indicators,imaging and pathological changes.Methods Clinical data,laboratory tests,imaging and pathological changes of IIM cases (n=85) and healthy controls (n=70) were enrolled.Clinical data included the classification,age,gender,course of the disease;laboratory tests including erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),creatine kinase (CK),creatine kinase isoenzyme-MB (CKMB),lactate dehydrogenase (LDH),hydroxybutyrate dehydrogenase (HBDH).The level of peripheral Th17,Treg cells and clinical indicators,laboratory tests,imaging and pathological changes were analyzed retrospectively.Since the data was disregarded from the normal distribution,the median four quantile method was used for statistical description.Two samples were compared with Mann-Whitney U test,and the correlation between variables was Spearman rank correlation analysis.Results ①) The levels of Th17 cells in the case group was not significantly different from that in the control group [6.18(3.42,13.65) cell/μl vs 7.42(5.02,11.13) cell/μl,P＞0.05],the levels of Treg cells in patients was significantly lower than that in the control group [21.25(12.48,35.67) cell/μl vs 36.95(30.37,47.12) cell/μl,P＜0.05],the ratio of Th17/Treg was also significantly higher than that in the control group [0.31(0.21,0.47) vs 0.18(0.14,0.31),P＜0.05].② Peripheral Treg cells levels were not correlated with ESR,CRP,CK-MB,LDH and HBDH (P＞0.05).Peripheral Treg cells levels were negatively correlated with CRP (r=-0.279,P＜0.05),but no correlated with ESR,CK-MB,LDH and HBDH (P＞0.05).③ According to the involvement of important organs,patients were classified into two groups:organ involvement group and non-organ involvement group.The levels of Treg cells in the organ involvement group was fewer than that in non-organ involvement group [16.54(8.84,27.34) cell/ul vs 24.87(14.44,43.37) cell/ul,P＜0.05],and the ratio of Th17/Treg in the organ involvement group was significantly higher than that in non-organ involvement group [0.41(0.29,0.68) vs 0.29(0.19,0.39),P＜0.05].④) Peripheral Th17 cells levels in patients with skeletal muscle inflammatory edema was significantly higher than that of non-inflammatory edema patients [10.70 (4.11,14.51) cell/μl vs 3.10 (1.27,5.15) cell/μl,Z=-2.460,P＜0.05].⑤ The levels of Th17,Treg cells and ratio of Th17/Treg did not correlate with pathological features of inflammatory infiltration (P＞0.05).Conclusion The absolute number of peripheral Treg cells decreases significantly in IIM,and correlates with CRP.Patients with organ involvement have fewer Treg cells,and there is imbalance between Th17 and Treg.When muscle MRI presents with inflammatory edema,patients may have high level of Th17 cells.Our results suggest that Treg cells may play an important role in the pathogenesis of IIM.