Objective:To analyze the protective effect of a respiratory syncytial virus (RSV) vaccine based on bacterial outer membrane vesicle (OMV) in mice.Methods:The pre-fusion protein (preF) of RSV was linked to the surface of OMV through the transmembrane protein cytolysin A (ClyA) to form the nanovaccine OMV-preF. The morphological characteristics of OMV and OMV-preF were observed under a transmission electron microscope. OMV-preF was intramuscularly injected into BALB/c mice and the elicited humoral and cellular immune responses were evaluated. The protective effect of OMV-preF was assessed by challenging the immunized mice with RSV Long strain. One-way analysis of variance and Tukey test were used for statistical analysis.Results:The results showed that preF was stably expressed in OMV, and both OMV-preF and OMV exhibited a double-layer vesicle structures under the microscope. OMV-preF could significantly activate the cellular and humoral immune responses in mice, causing a significant increase in CD8 + T cells and CD19 + B cells as well as a significant increase in the serum level of specific IgG. The neutralizing antibodies produced in the immunized mice could significantly inhibit the replication of RSV Long strain in vivo. Conclusions:The nanovaccine OMV-preF can induce high-level humoral and cellular immune responses, and the antibodies produced following immunization can effectively inhibit viral replication. This study provides a new strategy for RSV subunit vaccines.

Objective:To analyze the protective effect of a respiratory syncytial virus (RSV) vaccine based on bacterial outer membrane vesicle (OMV) in mice.Methods:The pre-fusion protein (preF) of RSV was linked to the surface of OMV through the transmembrane protein cytolysin A (ClyA) to form the nanovaccine OMV-preF. The morphological characteristics of OMV and OMV-preF were observed under a transmission electron microscope. OMV-preF was intramuscularly injected into BALB/c mice and the elicited humoral and cellular immune responses were evaluated. The protective effect of OMV-preF was assessed by challenging the immunized mice with RSV Long strain. One-way analysis of variance and Tukey test were used for statistical analysis.Results:The results showed that preF was stably expressed in OMV, and both OMV-preF and OMV exhibited a double-layer vesicle structures under the microscope. OMV-preF could significantly activate the cellular and humoral immune responses in mice, causing a significant increase in CD8 + T cells and CD19 + B cells as well as a significant increase in the serum level of specific IgG. The neutralizing antibodies produced in the immunized mice could significantly inhibit the replication of RSV Long strain in vivo. Conclusions:The nanovaccine OMV-preF can induce high-level humoral and cellular immune responses, and the antibodies produced following immunization can effectively inhibit viral replication. This study provides a new strategy for RSV subunit vaccines.

Objective To explore the value of combined detection of serum mitogen-activated protein kinase 1(MAPK1)and lysyl oxidase-like protein 2(LOXL2)in early diagnosis of cervical cancer.Methods A total of 218 patients with cervical lesions were selected as study group(103 ca-ses in cervical cancer group,115 cases in benign tumor group).Additionally,100 patients with cer-vical intraepithelial neoplasia grade Ⅱ were selected as precancerous lesion group,and 79 healthy in-dividuals undergoing physical examinations during the same period were selected as control group.Se-rum levels of MAPK1 and LOXL2 were measured in each group.Pearson correlation analysis was used to evaluate the correlations of serum MAPK1 and LOXL2 levels in patients with cervical cancer.Logistic regression analysis was performed to screen influencing factors for the occurrence of cervical cancer.Receiver operating characteristic(ROC)curves were plotted to assess the diagnostic efficacy of serum MAPK1 and LOXL2 for cervical cancer.Results Serum MAPK1 and LOXL2 levels in the study group were higher than those in the precancerous lesion group and the control group,and those in the precancerous lesion group were higher than those in the control group,with statistically signif-icant differences(P＜0.05).The proportion of patients with high-risk human papillomavirus(HPV)infection and serum MAPK1 and LOXL2 levels in the cervical cancer group were higher than those in the benign tumor group,with statistically significant differences(P＜0.05).Serum MAPK1 and LOXL2 levels in patients with stage Ⅲ to Ⅳ cervical cancer were higher than those in patients with stage Ⅰ to Ⅱ cervical cancer,with statistically significant differences(P＜0.05).Pearson correlation analysis showed a positive correlation between serum MAPK1 and LOXL2 levels in patients with cervical cancer(r=0.468,P＜0.001).Logistic regression analysis showed that high-risk HPV infection,MAPK1 and LOXL2 were all influencing factors for the occurrence of cervi-cal cancer(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)for com-bined diagnosis of serum MAPK1 and LOXL2 was 0.911,which was significantly greater than the AUCs for individual diagnoses(0.848 and 0.843,respectively).Conclusion Serum MAPK1 and LOXL2 levels in patients with cervical cancer are significantly upregulated,and the two indicators were positively correlated.High-risk HPV infection,serum MAPK1 and LOXL2 levels were influen-cing factors for the occurrence of cervical cancer.Combined detection of MAPK1 and LOXL2 levels is expected to assist in the diagnosis of cervical cancer.

Objective To determine the risk factors associated with early allograft dysfunction (EAD) after liver transplantation. Methods We retrospectively analyzed the records of 138 patients who underwent liver transplantation from January 2006 to October 2016 in our department. Transplant recipients were divided into two groups:those who met the diagnostic criteria of EAD (EAD group) and those who did not (non-EAD group). We compared blood flow into the transplanted livers and other clinical features between the two groups using univariate and multivariate analysis. Results Intraoperative portal vein flow (PVF) maximum was significantly different between the two groups. Multivariate analysis revealed that intraoperative PVF maximum ＜1 600 mL/min was the only independent risk factor for the occurrence of EAD after liver transplantation in this cohort. Conclusion Intraoperative PVF maximum ＜1 600 mL/min is an independent risk factor for the occurrence of EAD after liver transplantation. Measuring intraoperative blood flow into the transplanted liver in liver transplant recipients may help identify patients at risk for developing EAD.

Objective To summarize the characteristics and treatment methods of de novo malignancies in recipients after renal transplantation. Methods Clinical data of 43 patients with de novo malignancies after renal transplantation among 759 recipients were analyzed retrospectively. Characteristics, treatment methods and prognosis of the de novo malignancies after renal transplantation were summarized. Results The incidence of de novo malignancies in recipients after renal transplantation was 5.7％. The age of onset was (52±11) years old, and the de novo malignancies was diagnosed in 13-193 months with the median of 60 months after renal transplantation. The 43 patients with de novo malignancies included 9 cases of primary renal carcinoma, 7 cases of bladder carcinoma, 6 cases of lung carcinoma, 5 cases of lymphoma, 4 cases of colorectal carcinoma, 4 cases of mammary carcinoma, 2 cases of skin carcinoma, 1 case of adrenal carcinoma, 1 case of gastric carcinoma, 1 case of primary carcinoma of liver, 1 case of pancreatic carcinoma, 1 case of scalp angiosarcoma and 1 case of meningioma, and they were treated by surgical procedure, adjusting immunosuppressive therapy, radiotherapy or chemotherapy after diagnosed. The postoperative 1- and 5-year survival rates were 81％ and 63％, respectively. Conclusions The incidence of de novo malignancies in recipients after renal transplantation is higher than that in healthy subjects, and urological neoplasm is most common. Radical resection should be considered first, and antineoplastic combined therapy can be performed for the patients who cannot undergo surgery. Meanwhile, dosage of immunosuppressive agents can be reduced and medication regimens can be adjusted, thus effectively prolonging the survival time of patients.

Objective:To investigate the effectiveness of heart rate deceleration capacity (DC) measurement in predicting the car-diotoxicity of malignant tumor patients treated with epirubicin-based chemotherapy. Methods:The clinical medical records, including CK-MB and cTnI levels and dynamic electrocardiogram (ECG) parameters before and after each chemotherapy cycle, of 140 patients treated with epirubicin-based chemotherapy were analyzed. Patients were divided into the DC>4.5 ms group and the DC≤4.5 ms group based on the calculated DC values. The CK-MB and cTnI levels and the dynamic ECG parameters of the two groups were compared af-ter two and four cycles of chemotherapy. Results:Patients in the two groups exhibited no statistically significant difference in their rele-vant clinical and pathological data before receiving chemotherapy (P>0.05). However, after four cycles of chemotherapy, the DC≤4.5 ms group showed a significantly greater increase in serum CK-MB and cTnI concentrations over the pre-chemotherapy levels compared with the DC>4.5 ms group. After two and four cycles of chemotherapy, the DC≤4.5 ms group also exhibited a significantly greater in-crease in mean heart rate (beats/min) and supraventricular and ventricular arrhythmia counts (times/24 h) over the pre-chemotherapy values compared with the DC>4.5 ms group (P<0.05). After four cycles of chemotherapy, 23 cases showed abnormally elevated cTnI levels in the DC≤4.5 ms group. In this group, patients with elevated cTnI level exhibited no statistically significant difference in CK-MB and cTnI concentrations, mean heart rates, and supraventricular and ventricular arrhythmia counts compared with those with nor-mal cTnI level before chemotherapy (P>0.05). However, the DC values of patients with elevated cTnI were significantly lower than those with normal cTnI level (P<0.05). Conclusion:The risk of epirubicin-induced cardiotoxicity increased with decrease in DC value. The DC test was shown to be an effective predictor of the risk of epirubicin-induced cardiotoxicity.

Objective To compare the early renal function recovery after kidney transplant from donation after cardiac death (DCD) and brain death (DBD).Methods The Medline (1950-2011),Embase and Cochrane library database were searched and supplemented by review of conference proceedings and publication bibliographies.All original single institution studies reporting outcomes for DCD and DBD kidney transplant recipients were considered.Odds ratios (OR) and 95％ confidence intervals (CI) based on random effects models were calculated.Results Nine publications,all cohort studies,involving 2049 DCD and 5498 DBD recipients,were included.DCD recipients had 7.24 times increased odds of DGF (OR=7.24,95％ CI =3.86-13.58),and 4.97 times increased odds of PNF (95％ CI =3.77-6.55).Conclusion DCD renal transplantation is associated with higher risks of DGF and PNF.

Objective To summarize our experience in the liver transplantation from the donation after cardiac death (DCD).Methods The livers from three DCD donors (2 cases of brain trauma and 1 case of cerebral hemorrhage) were harvested according to the Guidelines for Donation after Cardiac Death in China.These grafts were orthotopically transplanted into three recipients including 2 cases of decompensative hepatic cirrhosis and 1 case of primary liver cancer.The warm ischemic time ranged from 7.5 to 10 min and the cold ischemic time was 4.5,8.2 and 6.5 h respectively.Postoperative immunosuppressive regimens included prednisone,FK506 and mycophenolate mofetil (MMF).Antibiotics and anticoagulatants were used accordingly.Results All of the 3 recipients obtained normal liver function within 3 weeks since the grafts were implanted without PNF,thrombosis and rejection.No postoperative complications occurred in 3 recipients during the follow-up period of 2 to 9 months with normal liver function.Conclusion The liver transplant from DCD donor showed good results in our center.Chinese group Ⅲ of DCD donor,UW score above the middle level and the short warm ischemic time are three keys ensuring the success of the liver transplant from DCD donors.

Objective To summarize the clinical experience of harvesting and using the kidneys from donation after cardiac death (DCD) donors.Methods Fourteen kidney transplantations were successfully performed on 14 patients with end-stage renal diseases.The kidneys were harvested from 7 volunteer donors (age 30～53 years) diagnosed with cardiac death,who were scored 19～23according to the University of Wisconsin donation after cardiac death evaluation.Primary diseases of the donors were cerebral hemorrhage,brain injury,ischemic cerebral vascular disease and brain tumor.Warm ischemia time ranged from 5 to 45 min,and cold ischemia time was 4.5 ～ 12.5 h.Results After transplantation,three patients had delayed graft function (DGF),one had primary non-function (PNF),and two patients developed acute rejection.In the patient with PNF,the transplanted kidney was removed one day after operation and the patient went back to hemodialysis.One patient with DGF was still in recovery with serum creatine 149 μmnol/L (within 3 months after operation).The above two cases both utilized the kidneys with 45 min of warm ischemia time.The rest 12 patients were discharged with normal renal function.Conclusion Under the condition of our country,kidneys strictly harvested from DCD donors can be used as one of the main sources of kidney grafts for kidney transplantation.

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.