Objective: To prepare an inactivated hepatitis A vaccine using a attenuated strain of hepatitis A virus (HAV) H2 and to analyze its immunizing dose, so as to provide the reference for development and production of inactivated hepatitis A vaccines. Methods: Human embryonic lung diploid cells (KMB17) were infected with attenuated HAV H2 strain to proliferate the virus, then the cells containing viruses were harvested, extracted and purified. The obtained virus concentrate was prepared into vaccine bulk and test vaccines with 1 280 EU/mL antigen content. Vaccine testing was carried out according to the inactivated hepatitis A vaccine standards specified in the Part Ⅲ of the Pharmacopoeia of the People's Republic of China (2020 edition). A total of 110 mice were randomly divided into 11 groups, including 5 dose groups (80, 160, 320, 640 and 1 280 EU/dose) of the test vaccine and the reference vaccine, as well as the adjuvant control group. Mice were immunized twice by intraperitoneal injection, their serum HAV antibodies were detected, and the geometric mean titer (GMT) and positive conversion rate of antibodies were analyzed to evaluate the immunising dose of the vaccine. Results: The antigen content and viral titer of the virus harvest solution were 5 120 EU/mL and 8.33 lgCCID50/mL, respectively. The removal rate of foreign protein reached 98.05% and the recovery rate of antigen was 66.25%. The test vaccine met the requirements of Part Ⅲ of the Pharmacopoeia of the People's Republic of China (2020 edition). The GMTs of HAV antibodies in the test vaccine and the reference vaccine dose groups after the second immunization were more than twice higher than those after the first immunization. Regardless of primary immunization or secondary immunization, the GMTs (log2) of HAV antibodies in the test vaccine groups with doses of 160 EU/dose and above were higher than those in the 80 EU/dose group (all P<0.05), while there was no statistically significant differences between the dose groups of 160 EU/dose and above (all P>0.05). The antibody positive conversion rate of 160 EU/dose and above of the test vaccine was 100.00% after the secondary immunization. Conclusions The inactivated hepatitis A vaccine of attenuated H2 strain tested in this study demonstrates strong immunogenicity in mice, suggesting its potential as a candidate vaccine. The preliminary analysis indicates an immunizing dose of 320 EU/dose for children and 640 EU/dose for adults.

Objective To explore the expression and significance of neutrophil extracellular traps(NETs)in peripheral blood of primary Sj?gren's syndrome(pSS)patients across different disease activity levels,and the predictive value of NETs and routine laboratory markers antithrombin Ⅲ(AT Ⅲ),alkaline phosphatase(ALP)and carbohydrate antigen 125(CA125)for pSS disease activity.Methods A total of 94 newly diagnosed pSS patients at the Affiliated Lianyungang Hospital of Xuzhou Medical University from October 2021 to December 2023 were categorized into active(n=49)and non-active(n=45)groups based on the European League Against Rheumatism(EULAR)Sj?gren's Syndrome disease activity index(ESSDAI).The levels of NETs biomarkers,namely serum myeloperoxidase(MPO)-DNA,were measured using ELISA.Laboratory routine indicators and MPO-DNA were integrated into multivariate Logistic regression to screen for independent influencing factors of pSS disease activity.Pearson's correlation was used to evaluate the relationship between MPO-DNA levels and ESSDAI scores.The efficacy of MPO-DNA alone or in combination with AT Ⅲ,ALP and CA125,for predictors of disease activity was evaluated using ROC curve.Results Serum MPO-DNA(23.884±3.494 μg/L),ALP(80.159±34.318 U/L)and CA125(20.300±16.560 U/ml)levels of active pSS patients were higher than those in the non-active patients(19.024±3.324 μ g/L,67.500±21.166U/L,13.200±6.340 U/ml),while AT Ⅲ(89.180±15.040 ng/ml)was lower than that in non-active patients(94.650±11.620 ng/ml),with significant differences(t=-7.921,-2.426,-2.925,2.094,all P＜0.05).Multivariate Logistic regression analysis showed laboratory indicator MPO-DNA,ALP and CA125 were independent risk factors,while AT Ⅲ was an independent protective factor(all P＜0.05).MPO-DNA was positively correlated with ESSDAI scores(r=0.602,P＜0.01).The AUC(95％CI)of the combination of ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.711(0.612～0.810).The AUC(95％CI)of MPO-DNA alone for predicting disease activity in pSS was 0.837(0.758～0.915),and the AUC(95％CI)of combination of MPO-DNA,ALP,CA125 and AT Ⅲ for predicting disease activity in pSS was 0.866(0.797～0.935),showing an improving in the predictive value.Conclusion The involvement of NETs in the occurrence and expression levels of pSS is related to its disease activity.NETs combined with ALP,CA125 and AT Ⅲ have effective diagnostic performance for the disease activity of pSS.This tool can serve as a biological indicator for predicting the disease activity of pSS.

Objective:To compare the prognoses of salvage liver transplantation fulfilling the Criteria of Milan, University of California San Francisco(UCSF)and Hangzhou.Methods:Clinical data were retrospectively reviewed for 256 patients with recurrent hepatocellular carcinoma(HCC)undergoing donation after citizen death(DCD)liver transplantation(LT)from January 2015 to October 2019.They were divided into two groups of primary(PLT, n=175)and salvage(SLT, n=81). General profiles, tumor pathological characteristics and postoperative complications of two groups were compared by T-test, rank-sum or χ2 test.Kaplan-Meier method and Log rank test were employed for comparing overall survival rate(OS)and recurrence-free survival rate(RFS)between two groups.In SLT group, 31 cases fulfilled Milan criteria, 45 cases UCSF criteria and 69 cases Hangzhou criteria.OS/RFS of three groups were compared.According to there was downstaging or bridging treatment pre-LT, SLT group was divided into downstaging group(n=32)and non-downstaging group(n=49). OS/RFS of two groups were compared.According to the Rescit1.1 criteria, downstaging group were divided into remission group(n=14)and non-remission group(n=18)and OS/RFS of two groups were compared. Results:The operative durations of PLT and SLT groups were(439.5±74.9)and(475.1±83.4)min respectively.There was significant inter-group difference( P<0.05); However, no significant inter-group difference existed in amount of intraoperative bleeding, blood transfusion, postoperative hospital stay or incidence of postoperative complications(all P>0.05). No significant difference existed in OS/RFS between PLT and SLT groups( P>0.05). No significant difference existed in OS at 1/3/5 years post-SLT among Milan, UCSF and Hangzhou criteria groups(all P>0.05); However, RFS in Milan criteria group at 1/3/5 years post-SLT were 93.5%, 81.7% and 81.7% respectively.They were significantly higher than 68.9%, 59.7% and 59.7% in UCSF criteria group and 78.3%, 58.8% and 55.5% in Hangzhou criteria group(all P<0.05). For patients on downstaging therapy, OS in the Remission group at 1, 3 and 5 years post-SLT were 100%, 73% and 73% respectively, which was significantly higher than 83.3%, 49.4% and 0 in non-Remission group( P=0.042). RFS in the Remission group at 1, 3 and 5 years post-SLT were 100%, 62.5% and 46.9% respectively, which was significantly higher than 52.9%, 0 and 0 in no-Remission group( P=0.001). Conclusions:The survival outcome of SLT recipients is similar to that of PLT recipients.The overall survival of SLT recipients shows no significant difference between Milan, UCSF and Hangzhou criteria.However, SLT recipients fulfilling Milan criteria have the longest recurrence-free time.The prognosis of patients with remission after preoperative descending treatment is superior to that of patients without remission.

Objective:To explore the effect of pre-transplant immunotherapy on the prognosis of transplant recipients with hepatocellular carcinoma(HCC).Methods:From June 2018 to September 2021, retrospective analysis was conducted for clinical data of 19 HCC-liver transplant recipients receiving pre-transplant immunotherapy in affiliated Huashan Hospital of Fudan University. Pre-transplant immunotherapy regimen, adverse reactions, post-transplant acute rejection, tumor recurrence and metastasis and other complications were recorded. According to the preoperative tumor imaging and the changes of alpha-fetoprotein level, tumor change during recipient waiting period was judged by the mRECIST standard. According to whether or not there was partial tumor remission, they were divided into two groups of non-remission( n=13)and remission( n=6). Postoperative conditions of two groups were compared. Kaplan-Meier method was used for calculating the survival rate of recipients after transplantation and survival curve and Log-rank test utilized for comparing the recurrence-free and overall survival rates of recipients at 1 and 2 years post-operation. Results:A total of 19 liver transplant recipients received immunotherapy plus targeted and transcatheter arterial chemoembolization(TACE) before transplant. In non-remission group, tumor was stable( n=9)and progressive( n=4); 6 cases in remission group had tumor partial remission. Two recipients in non-remission group were pathologically confirmed by liver biopsy to have acute rejection(2/19, 10.5%)and both recovered after glucocorticoid + rATG and glucocorticoid therapy. In non-remission group, 2 patients died from septic shock post-operation. Among 3 patients of tumor recurrence and metastasis post-operation, 2 cases survived with tumor and 1 died after tumor recurrence and metastasis. In remission group( n=6), none had postoperative tumor recurrence and metastasis. The recurrence-free survival rates of non-remission group recipients at 1 and 2 years post-operation were 76.9% and 76.9% and recurrence-free survival rates in remission group were 100% and 100% respectively and inter-group difference in RFS was not statistically significant( χ2=1.468, P=0.226). The overall survival rates of recipients in non-remission group at 1 and 2 years post-operation were 76.9% and 76.9% respectively. And recipients in remission group were 100% and 100% respectively and no statistically significant inter-group difference existed in OS( χ2=1.292, P=0.256). Conclusions:Without a significantly higher risk of acute rejection after transplant, immunotherapy may be an effective option for bridging treatment before liver transplantation for HCC. And it remains necessary to expand the sample size for verifications and supports.

Objective:To analysis the risk factors and safety of administration of norepinephrine (NE) via peripheral vein line (PVL) in patients with septic shock.Methods:A single-center retrospective study was conducted. According to the Lanzhou University Second Hospital information system (HIS) and nursing adverse events report cards, patients with septic shock administrated with NE via PVL to correct the hypotension from January 1st 2015 to December 31st, 2019 were enrolled. The patients' general information, placement location of peripheral venous catheter and venousneedle type, characteristics of NE usage and patient general condition when extravasation occurred were collected. The univariate analysis and Logistic regression were used to analyze risk factors associated with extravasation. Also, the receiver operator characteristic curve (ROC curve) was drawn, and the predictive value of risk factors for extravasation was analyzed.Results:A total of 1 022 cases with NE were enrolled. After a preliminary screening, a total of 910 cases with NE were used to correct low blood pressure, including 116 cases of peripheral venous infusion. The average age was (52.91±18.69) years old, with majority of female (77 cases, 66.4%). Basic diseases were mainly chronic obstructive pulmonary disease (COPD, 100 cases, 86.2%), followed by hypertension(91 cases, 78.4%), coronary heart disease (87 cases, 75.0%), type 2 diabetes (74 cases, 63.8%) respectively, the primary disease was septic shock in 109 cases (94.0%). A total of 147 peripheral venous catheters were inserted, and the most common site of puncture was the forearm [78.9% (116/147)], followed by the hand [12.2% (18/147)] and the median cubital vein [8.8% (13/147)]. 89.9% of the needles were 20 G in diameter, and 75 cases (64.7%) were converted to central venous catheters (CVC) during subsequent treatment due to continuous infusion of NE. Six patients (5.2%) had extravasation, the median time of extravasation was 29 (23-39) hours, and the median time of NE was 23 (11-53) hours, including 2 patients with an infusion concentration of 60 mg/L and 4 patients with an infusion concentration of 120 mg/L.The infusion speed was 0.5-1.0 μg·kg -1·min -1, and the average speed of infusion was (0.75±0.04) μg·kg -1·min -1 when extravasation. Univariate and binary Logistic regression analysis showed that the risk factors related to the occurrence of extravasation included: ① patient factors: the presence of basic diseases, hypertension [odds ratio ( OR) = 3.11, 95% confidence interval (95% CI) was 3.09-3.12, P = 0.001] and edema ( OR = 1.79, 95% CI was 1.32-2.99, P = 0.032). ② Factors of infusion fluid itself: long-term (> 24 hours) infusion ( OR = 2.91, 95% CI was 1.04-5.96, P = 0.040), infusion concentration > 60 mg/L ( OR = 1.88, 95% CI was 1.32-3.99, P = 0.024), infusion speed > 0.3 μg·kg -1·min -1 ( OR = 2.43, 95% CI was 2.38-2.51, P = 0.029) and diameter of needles < 20 G ( OR = 3.11, 95% CI was 3.09-3.22, P = 0.033).③ Medical personnel factors: lack of observation and assessment ( OR = 1.09, 95% CI was 1.03-6.77, P = 0.043). The ROC curve analysis showed that: edema, long-term infusion (> 24 hours), infusion rate > 0.3 μg·kg -1·min -1and diameter of needles < 20 G had a certain predictive value for extravasation of NE through peripheral venous infusion in patients with septic shock, the area under ROC curve (AUC) was 0.610, 0.762, 0.672, 0.629, 95% CI was 0.508-0.713, 0.675-0.849, 0.571-0.772, 0.525-0.732, and P values were 0.044, 0.000, 0.002, 0.019, respectively. Conclusions:Hypertension, edema, long-term infusion (> 24 hours), infusion concentration > 60 mg/L, infusion speed > 0.3 μg·kg -1·min -1, diameter of needle < 20 G, and lack of observation and evaluation by medical staff regularly were risk factors affecting the safety of peripheral intravenous NE in patients with septic shock. Peripheral NE should be avoided in the presence of the above risk factors.

Objective:To explore the relationship between CD24 expression in preoperative peripheral blood as well as cancer tissue and clinical parameters and prognosis in patients with hepatocellular carcinoma (HCC) after liver transplantation (LT).Methods:From November 2018 to November 2019, clinical data were collected for 65 HCC patients and 41 patients with benign liver disease.The preoperative peripheral blood level of CD24 was detected by enzyme-linked immunosorbent assay (ELISA) and the expression of CD24 in cancerous foci and adjacent tissues examined by immunohistochemistry.Kaplan-Meier survival curves of differential CD24 expression were plotted and survival differences compared by Log-rank method.One-way ANOVA was utilized for examining the relationship between the expression level of CD24 and various clinicopathological parameters and multivariate Cox analysis for screening independent risk factors affecting patient prognosis.Results:The concentration of CD24 in preoperative peripheral blood (p-CD24) of HCC patients (6.51±2.33 μg/L) was significantly higher than that of patients with benign liver disease (4.10±0.91) μg/L, P<0.05.The positive rate of CD24 was obviously higher in cancerous tissues than that in adjacent tissues (87.7% vs. 4.6%, P<0.05). The peripheral blood level of CD24 was positively correlated with the expression intensity of CD24 in tumor tissues (t-CD24, r=0.570, P<0.001). The expression of CD24 (both in blood and cancer foci) was significantly correlated with preoperative level of gamma-glutamyl transferase (GGT), maximal tumor diameter, microvascular invasion, portal vein tumor thrombus, vessel carcinoma embolus and satellite focus ( P<0.05). The expression of CD24 in patients exceeding the Milan/UCSF criteria was higher than those fulfilling the criteria ( P<0.005). Patients with a higher expression of CD24 had worse overall survival and recurrence-free survival rates as compared to those a lower expression of CD24 ( P<0.05). Multivariate Cox analysis indicated that t-CD24 [OS: HR=3.661(1.005-13.333)], P=0.049; recurrence-free survival (RFS): [HR=4.331(1.887-9.942), P=0.001] and preoperative level of alpha fetoprotein (AFP) [OS: HR=4.900(1.590-15.097), P=0.006]; RFS: [HR=3.414(1.614-7.221), P=0.001] were independent risk factors for overall survival and recurrence-free survival in HCC patients undergoing LT. Conclusions:The preoperative peripheral blood level of CD24 in HCC patients undergoing LT indirectly reflects the expression of CD24 in cancerous tissues to a certain extent.And the expression of CD24 in cancerous tissue is one of the independent risk factors affecting OS and RFS of LT patients.

【Objective】 To investigate the removal efficacy of inflammatory cytokines and blood compatibility of modified PBTNF. 【Methods】 Acrylic acid (AA) was firstly UV-grafted onto the surface of PBTNF to negatively charge the surface of the material. Subsequently, the three positively charged polyelectrolytes, DA, PEI, and CS were respectively electrostatic self-assembled with GO on the surface of PBTNF, forming two layers of film with GO as the outer layer: PBTNF-(DA/GO)2, PBTNF-(PEI/GO)2, PBTNF-(CS/GO)2. 【Results】 Scanning electron microscopy results showed that compared with the PBTNF grafted with AA, the adhesion of particles was observed on the surface of the three modified materials, and the photo shows that the color of the material surface was deepened after electrostatic self-assembly. The results of wettability showed that the surface hydrophilicity was significantly improved, indicating that the electrostatic self-assembled membrane was successfully immobilized on the surface of PBTNF. The removal efficiency (%) of IL-1β for PBTNF-(DA/GO)2, PBTNF-(PEI/GO)2 and PBTNF-(CS/GO)2 were 69.00±7.36 vs -2.35±2.69 vs -1.59±3.26 (P<0.05). The removal efficiency of IL-6 (%) were 40.15±1.86 vs -13.46±5.72 vs -1.21±3.41 (P<0.05). The removal efficiency of IL-8 (%) were 96.90±0.97 vs 17.84±11.74 vs 43.68±17.38 (P<0.05). The removal efficiency of TNF-α (%) was 44.46±2.50 vs 14.90±7.12 vs 20.64±1.22 (P<0.05). Plasma protein adsorption results (total protein, immunoglobulin G, albumin) and red blood cell deformability index showed that there was no statistical difference among the three modified PBTNFs and the control group (P>0.05). Although the red blood cell osmotic fragility (g/L) of the three modified PBTNFs is higher than that of the former: control group vs PBTNF-(DA/GO)2 vs PBTNF-(PEI/GO)2 vs PBTNF-(CS/GO)2: 4.39±0.05 vs 4.62±0.02 vs 4.48±0.03 vs 4.90±0.03 (P<0.05), the hemolysis rate (%) of them were all less than 5%, and PBTNF-(DA/GO)2 performed the lowest hemolysis rate which was (0.03±0.01)% (compared with PBTNF-(PEI/GO)2, P<0.05). The coagulation function test results showed that compared with the control group, the fibrinogen (g/L) of the three modified PBTNFs had no statistical difference (P>0.05); the activated partial thrombo plastin time (S) slightly extended, but all within the normal range of clinical standard; and the prothrombin time (S) of PBTNF-(CS/GO)2 was prolonged(P<0.05). 【Conclusion】 Among the three positively charged polyelectrolytes, including DA, PEI, and CS, PBTNF-(DA/GO)2 performed the best removal rate of inflammatory cytokines, and the blood compatibility evaluation results showed that PBTNF-(DA/GO)2 had no significant effect on red blood cells and coagulation function. Consequently, in the study of inflammatory cytokines adsorption, DA is expected to be the optimal polyelectrolyte assembling with GO for further research.

Objective: To investigate the value of Baduanjin combined with balance pad exercise in improving the balance ability and motor function of elderly Parkinsonundefineds patients. Methods: Totally 120 elderly patients with Parkinsonundefineds disease admitted in our hospital from March 2017 to March 2018 were divided into two groups according to different exercise methods: the balance pad training group (60 cases) and the Baduanjin combined balance pad group (60 cases). After 2 months of continuous exercise, Fugl-Meyer lower limb motor function scale and Berg Balance Scale (BBS) were used to evaluate the recovery of lower limb motor ability and the changes of somatic balance control. Before training and 2 months after training, Parkinsonundefineds Scale (UPDRS) was used to evaluate the motor impairment of the patients. Results: There were no significant differences between the two groups in the Fugl-Meyer score, BBS score, 6mWD and UPDRS scores before training (P>0.05). After 2 months of training, the BBS score of the observation group was (45.5±4.0) points and Fugl-Meyer score. (25.5±3.2) points and 6mWD scores (362.2±44.9) points, the control group was (42.3±3.8) points, (22.7±3.6) points, (337.3±51.0) points, the difference was statistically significant (t=4.493, 4.503, 2.839, P<0.05). After 2 months of training, the UPDRSII and III scores of the observation group were (11.8±3.0) points and (15.8±1.9) points, respectively, which were significantly better than the control group (14.7±2.6) points, (17.2 ± 1.4) points, the difference was statistically significant (t=5.658, 4.595, P<0.05).; the observation group patients had a fall rate of 16.67% (10/60), no fracture cases, the control group was 41.67% (25/60), 6.67% (4/60), the difference was statistically significant (χ²=9.076,-, P<0.05). Conclusion Bazuanjin combined with balance pad training can significantly improve lower limb motor function, restore gait, improve body balance control and improve quality of life in elderly patients with Parkinsonundefineds disease.

Objective To explore the clinical features and risk factors associated with intrahepatic and hilar cholangiocarcinoma after liver transplantation .Methods Retrospective analysis of clinical data was performed for 20 hospitalized patients with intrahepatic and hilar cholangiocarcinoma from June 25 ,2014 to October 31 ,2018 .Treatments and follow-up outcomes were analyzed .The survival rate was calculated by the Kaplan-Meier method and the survival curve plotted .Cox regression model was employed for analyzing the prognostic factors .Results The cumulative recurrence rate of patients with AJCC stage Ⅰ /Ⅱ was significantly lower than that in AJCC stage Ⅲ/Ⅳ .And the cumulative recurrence rate of stageⅠ/Ⅱ Patients was 0 and that of stage Ⅲ/Ⅳ 76％ (P=0 .042) .Cox regression model showed that CA19-9 was the only prognostic factor .An elevated level of CA19-9 was associated with high recurrence post-transplantation (HR=1 .001;95％ CI:1 .000～1 .001;P=0 .035) .Conclusions During progressive stage ,the recurrence rate is higher with a worse prognosis .And an elevation of CA19-9 is an independent poor prognostic factor after intrahepatic and hilar cholangiocarcinoma transplantation .

Objective: To explore the clinical efficacies of applying aged marginal donor liver. Methods: From January 2015 to June 2018, clinical data were retrospectively analyzed for 199 adult liver transplantation donors and recipients. They were divided into two groups of aged (≥60 years) and appropriate age (<60 years). The prognosis of two groups was compared after a follow-up period of 1 year. And the aged group was further assigned into lower and higher fat infiltration groups according to the degree of fat infiltration in donor liver and compared the prognosis of two groups. Results: No significant differences existed in initial, peak value and recovery time of transaminase (AST/ALT), peak value and recovery time of total bilirubin, glutamyl transpeptidase, alkaline phosphatase, international normalized ratio (INR), peak value of lactate, postoperative hospital stay, graft dysfunction, biliary/vascular complications, acute/chronic rejection or graft survival rate between aged and appropriate age groups post-transplantation. The aged group was further divided into lower and higher fat infiltration groups according to the fat infiltration rate (<20%, ≥20%). And significant inter-group differences existed in peak value and recovery time of AST/ALT, peak value of total bilirubin, glutamyl transpeptidase, lactate, postoperative hospital stay and graft dysfunction. The above parameters were significantly worse in higher fat infiltration group. Also the rejection rate was higher in high group at 1 year post-operation and no significant inter-group difference existed in biliary/vascular complications. In higher group, 4 patients showed graft dysfunctions during perioperative period. Two of them were discharged successfully after secondary transplantation and another 2 patients died. Conclusions On the premise of comprehensive evaluations of donor liver status and reasonable matching of recipients, aged marginal donor liver can be safely applied with excellent clinical outcomes. Severe fatty donor liver should be employed with caution. Hypertensive drugs, high serum sodium and long period of cold ischemia are also important influencing factors for aged donors.