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OBJECTIVES: To investigate the influencing factors of the image quality of shear wave elastography (SWE) performed using a high-frequency probe and its reproducibility for renal allografts. METHODS: A total of 211 patients with transplanted kidneys who underwent SWE examination performed using high-frequency or low-frequency probes were recruited for the study. The reproducibility of inter- and intraobserver agreements were analysed by using the intraclass correlation coefficient (ICC). According to the colour filling of the area of interest and imaging noise when conducting SWE, the image quality was classified as three grades: “good”, “common”, and “poor”. A logistic regression was used to analyse the independent factors for SWE quality. RESULTS: In the comparative analysis, high frequency, transection measurement and middle pole were selected as the appropriate measurement methods. Regarding reproducibility, the ICCs) of the intra- and interobserver agreements were 0.85 and 0.77, respectively. Multivariate analysis indicated that only the skin allograft distance and kidney width were independent variables for SWE quality. In the subgroup analysis of the skin-allograft distance, the “good” and “common” rates of images decreased as the distance increased, but the CV (coefficients of variation) showed the opposite trend. The SWE quality of kidney width

BACKGROUND: Registration of three-dimensional contrast-enhanced ultrasound fusion imaging (3DCEUS-FI) is time-consuming to obtain high success rate. OBJECTIVE: To investigate the influence factors on registration success rate of 3DCEUS-FI. METHODS: Water tank phantoms were made to obtain mimicked pre- and post- radiofrequency ablation three-dimensional contrast-enhanced ultrasound (3DCEUS) and CT images. Orthogonal trials were designed according to factors including size, depth, enhancement level of mimicked tumor, diameter and number of mimicked adjacent vessels. Mimicked pre- and post-RFA 3DCEUS images of 72 trials were fused to assess ablative margin (AM) by two radiologists. With CT images as standard, 3DCEUS-FI accuracy was considered as the consistency of AM evaluation. The inter-observer agreement and the influence factors on registration success rates were analyzed. RESULTS: The intraclass correlation coefficient (ICC) for the consistency of AM evaluation between CT and 3DCEUS-FI in x-axis, y-axis or z-axis was 0.840∼0.948 (P

This study aimed to develop and validate a combined nomogram model based on deep learning (DL) contrast-enhanced ultrasound (CEUS) and clinical factors to preoperatively predict the aggressiveness of pancreatic neuroendocrine neoplasms (PNENs).In this retrospective study, consecutive patients with histologically proven PNENs underwent CEUS examination at the initial work-up between January 2010 and October 2020. Patients were randomly allocated to the training and test sets. Typical sonographic and enhanced images of PNENs were selected to fine-tune the SE-ResNeXt-50 network. A combined nomogram model was developed by incorporating the DL predictive probability with clinical factors using multivariate logistic regression analysis. The utility of the proposed model was evaluated using receiver operator characteristic, calibration, and decision curve analysis.A total of 104 patients were evaluated, including 80 (mean age ± standard deviation, 47 years ± 12; 56 males) in the training set and 24 (50 years ± 12; 14 males) in the test set. The DL model displayed effective image recognition with an AUC of 0.81 (95%CI: 0.62-1.00) in the test set. The combined nomogram model that incorporated independent clinical risk factors, such as tumor size, arterial enhancement level, and DL predictive probability, showed strong discrimination, with an AUC of 0.85 (95%CI: 0.69-1.00) in the test set with good calibration. Decision curve analysis verified the clinical usefulness of the combined nomogram.The combined nomogram model could serve as a preoperative, noninvasive, and precise evaluation tool to differentiate aggressive and non-aggressive PNENs.• Tumor size (odds ratio [OR], 1.58; p = 0.02), arterial enhancement level (OR, 0.04; p = 0.008), and deep learning predictive probability (OR, 288.46; p0.001) independently predicted aggressiveness of pancreatic neuroendocrine neoplasms preoperatively. • The combined model predicted aggressiveness better than the clinical model (AUC: 0.97 vs. 0.87, p = 0.009), achieving AUC values of 0.97 and 0.85 in the training set and the test set, respectively.

This pilot clinical study evaluated primarily the efficacy of feeding vessel ablation (FVA) in the treatment of hepatocellular carcinoma (HCC) located at the liver marginal angle (LMA). Nine patients with nine unresectable HCC lesions were prospectively included in this study. The target tumors (mean: 3.0 cm, interquartile range: 2.4-3.6 cm) were located at the LMA (segment 2/3/6) and adjacent to the gastrointestinal tract. Artificial ascites was attempted and failed. Multimode ultrasound technologies, including 2-D and real-time 3-D contrast-enhanced ultrasound, were used to identify the morphology and structure of the feeding vessels for the target tumors. During the treatment, a unipolar cool-tip electrode was used to ablate the feeding vessels, and the target ablation point was set in subsegmental or more distal vessels to induce a downstream ischemia region. Therapeutic outcomes were assessed after FVA, including the rates of technical success, tumor response, local tumor progression (LTP), overall survival (OS) and major complications. Cumulative LTP and OS were estimated with the Kaplan-Meier method. The technical success rate determined immediately after radiofrequency ablation was 7 of 9 (77.8%). Complete response (CR) was achieved in 7 of 7 tumors (100%) at the 1-mo evaluation. During a median follow-up period of 15.6 mo (range: 4.3-53.3 mo), CR remained in 6 of 7 tumors (85.7%), with LTP observed in 1 of 7 tumors (14.3%) 4.7 mo after treatment. The cumulative 1-, 3- and 5-y LTP-free rates were all 83.3%, and the cumulative 1-, 3- and 5-y OS rates were 42.9%, 28.6% and 0%, respectively. No major complications occurred. We concluded that FVA can induce subsegmental devascularization and has the potential to serve as an effective and safe alternative technique for local control of unresectable HCC located at the LMA when artificial ascites fails.

To retrospectively assess the diagnostic performance of grey-scale ultrasound (US) characteristics and gamma-glutamyl transpeptidase (GGT) alone or combined in distinguishing biliary atresia (BA) from other cholestasis diseases in infants younger than 30 days. Between January 2012 and October 2020, the demographic characteristics, laboratory results and US characteristics of 35 BA and 52 non-BA infants younger than 30 days were retrospectively evaluated. Areas under the receiver operating characteristic curves (AUCs) were used to estimate the probability of predicting BA, which were compared by DeLong test. The diagnostic performance of gallbladder classification in identifying BA was higher than that of fibrotic cord thickness (AUC 0.900 vs. 0.771, P = 0.03). With the cutoff level of 188 IU/L, serum GGT had a sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 77.1%, 69.2%, 62.8%, and 81.8%, respectively. Combined with gallbladder classification and GGT, the sensitivity, specificity, PPV, NPV and accuracy were 100.0%, 63.5%, 64.8%, 100.0% and 78.2%. Gallbladder classification was more valuable than fibrotic cord thickness in the diagnosis of BA among infants less than 30 days. Combined with gallbladder classification and GGT, the sensitivity for the diagnosis of BA can reach 100.0%.

Purpose To compare the efficacy of radiofrequency (RF) ablation after transarterial chemoembolization within or beyond 30 days for medium-large or multiple recurrent hepatocellular carcinomas (HCCs). Materials and Methods In this single-center retrospective study conducted from 2007 through 2015, 135 patients with a single recurrent HCC (>3 cm) or multiple (2–5 tumors) recurrent HCCs underwent transarterial chemoembolization plus RF ablation. A total of 62 patients underwent RF ablation after transarterial chemoembolization within 30 days (sequential group) and 73 patients underwent RF ablation after transarterial chemoembolization beyond 30 days (delayed group). Outcomes of interests included overall survival (OS), progression-free survival (PFS), and complete response (CR) rate. Results The median OS and PFS were 49.8 and 38.0 months for sequential group, and 31.0 and 11.6 months for the delayed group. The sequential group experienced significantly better OS (hazard ratio [HR]: 0.517; P = .002) and PFS (HR, 0.621; P = .021). Among patients with multiple tumors or a single tumor >5 cm, the sequential group still had significantly longer OS (P = .022; P = .018, respectively) and PFS (P = 0.042; P = .036, respectively) than the delayed group, although no significant differences were observed among patients with solitary 3- to 5-cm tumors (P = .138; P = .803, respectively). The sequential group had a significantly better CR rate than the delayed group (85.4% vs. 68.5%, respectively; P = .035). Significant predictors of OS and PFS included maximum tumor size, number of tumors, and time interval between transarterial chemoembolization and RF ablation. Conclusions Transarterial chemoembolization plus sequential RF ablation within 30 days was more effective for recurrent HCCs than transarterial chemoembolization plus delayed RF ablation. The time interval within 30 days is required for treating large or multiple HCCs but may not be necessary for solitary medium-sized HCC.

Purpose: This study aims to complete a detailed record of the clinical characteristics and treatment of HCC patients with post-ablation infection and evaluate the infections on recurrence-free survival (RFS) and overall survival (OS) among patients receiving ultrasound-guided thermal ablation. Methods: 3117 patients with liver tumors receiving thermal ablation from January 2010 to December 2021 were analyzed. A total of 49 patients with infectious complications after thermal ablation were selected as the infection group. A total of 49 patients without postoperative infection were randomly selected among those who underwent ablation within three days before or after the treatment date of the infection group as the control group. The clinical characteristics of both groups were analyzed by an independent sample t-test and chi-square test. A log-rank test was performed to compare the RFS and OS data. A multivariate Cox regression model was employed to identify prognostic factors influencing RFS and OS. Subgroup analyses of mild and severe infections were conducted to explore the infection-related situation further. Results: Between mild and severe infection groups, there were statistically significant differences in the infection position (p = 0.043), positive rate of body fluid culture (p = 0.002), proportion of catheter drainage (p = 0.017), use of advanced antibiotics (p = 0.006), and outcome (p = 0.00). The Kaplan–Meier survival analysis revealed that postoperative infection was significantly correlated with tumor recurrence (p = 0.028), and severe infection was significantly associated with overall survival (p = 0.049). The cox model showed that postoperative infection was an independent variable for RFS deterioration (HR = 1.724, 95% CI: 1.038–2.862, p = 0.035). Conclusions: Postoperative infection among patients receiving ultrasound-guided thermal ablation adversely affected tumor progression. In addition, empirical antibiotics and catheterization to reduce pressure inside the lesion should be utilized to minimize symptoms in patients with postoperative infection.

To investigate the feasibility of assessing the ablative margin (AM) of radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC) with 3-D contrast-enhanced ultrasound fusion imaging (3-DCEUS-FI), pre- and post-RFA 3-DCEUS images of 84 patients with HCC were fused for two radiologists to independently assess the AMs. The success rate, duration and influencing factors for registration; inter-observer agreement for AM classification; and local tumor progression (LTP) rate were evaluated. The success rate of the automatic registration (AR), which was completed within 4-12 s, was 57.1% (48/84). The duration and success rate of the interactive registration (IR) were 4.2 ± 1.8 min and 91.7% (77/84) for radiologist A and 4.8 ± 2.1 min and 91.7% (77/84) for radiologist B, respectively. The multivariate analysis demonstrated that the pre-RFA image quality, number of vessels (≥3 mm) and presence of acoustic shadow were independent factors for AR (p0.05), while the number of vessels was an independent factor for IR (p = 0.001). The agreement between observers was excellent (kappa = 0.914). LTP rate was significantly higher for AMs5 mm than for AMs ≥5 mm (p = 0.024). Quantitatively evaluating the AM immediately after RFA for HCC with 3-DCEUS-FI was feasible.

Objectives The purpose of this study was to assess the characterization and usefulness of contrast-enhanced ultrasound (CEUS) for diagnosing solid pseudopapillary tumors of the pancreas (SPTP) and compare the enhancement patterns with contrast-enhanced computed tomography (CECT). Methods Forty-three SPTP lesions proved by pathologic findings in 42 patients examined with CEUS and CECT were included in this study. The enhancement characteristics and typical CEUS features of the tumors were investigated. These characteristics were compared according to lesion sizes. The enhancement patterns of CEUS were compared with CECT. Results The most common enhancement levels of SPTP in the early phase and late phase for CEUS were isoenhancement (19 of 43 [44.2%]) and hypoenhancement (32 of 43 [74.4%]), respectively. The 4 most common enhancement patterns were hypo-hypo (16 of 43 [37.2%]), iso-iso (11 of 43 [25.6%]), hyper-hypo (8 of 43 [18.6%]), and iso-hypo (8 of 43 [18.6%]) enhancement. For the 43 SPTP lesions, typical CEUS features such as lesion membrane, intralesional vessel, and intralesional compartmentalization enhancements were detected in 30 (69.8%), 27 (62.8%), and 10 (23.2%) cases. Compared with CECT, isoenhancement during the early phase and hypoenhancement during the late phase were the most common imaging characteristics of CEUS. Conclusions Lesion membrane, intralesional vessel, and intralesional compartmentalization enhancements are typical CEUS features of SPTP, especially for large lesions (lesion size ≥3.0 cm). Isoenhancement during the early phase and hypoenhancement during the late phase are the most common imaging characteristics of CEUS and CECT, making CEUS a viable alternative diagnostic method that is noninvasive.

Background The prognosis of adrenal metastases (AM) from hepatocellular carcinoma (HCC) with surgical contraindication was poor. This study evaluated the feasibility, safety and treatment efficacy of percutaneous ultrasound (US)-guided radiofrequency ablation (RFA) for the local treatment of AM originated from HCC. Methods A retrospective study was carried out on 22 patients (21 male and 1 female, mean age, 53.0 ± 13.0 years) who had single AM (mean diameter, 4.0 ± 1.8 cm, range, 1.7–8.0 cm) originated from HCC and received US-guided percutaneous RFA at our institution. The diagnosis was established on typical radiologic findings. The primary technical success was defined as the tumour being completely ablated in the first RFA session. The secondary technical success was defined as tumour residual left from the first ablation was completely ablated by a second ablation session. Local tumour progression (LTP) and overall survival (OS) were estimated by using Kaplan-Meier analysis. Results A total of 25 ablation sessions were performed. The primary technical success and the secondary technical success were 77.3% (17 of 22) and 86.4% (19 of 22), respectively, with the major complication rate at 4.5% (1 of 22). The median follow-up period after RFA was 10 months (3–55 months). During the follow-up period, five patients were detected LTP. The LTP at 3, 6, and 12 months were 15.8, 26.3, and 26.3%, respectively. Nine patients died of distant extra-adrenal metastases and another five of liver failure due to HCC. The OS at 6, 12, 24 months after RFA for AM were at 79.7, 52.6, and 32.9%, respectively. Conclusion Percutaneous US-guided RFA in the treatment of AM originated from HCC is feasible, safe and effective.

OBJECTIVES The aim of this study was to evaluate the influence of fasting time on the ultrasound identification and exclusion of biliary atresia in jaundiced infants through the use of the gallbladder classification scheme and to test the value of the gallbladder classification scheme in the diagnosis of biliary atresia in inexperienced individuals. METHODS A total of 188 jaundiced infants were enrolled in this study. All patients received detailed abdominal sonograms. Diagnoses were confirmed via liver biopsy, surgical findings, or follow-up. Infants were grouped into either the fasting group (fasting time ≥ 4 h) or the nonfasting group (fasting time

To retrospectively assess the value of the combination of conventional ultrasound and shear-wave elastography (SWE) in evaluating the segmental heterogeneity of liver fibrosis in biliary atresia (BA) patients after Kasai portoenterostomy. A total of 35 BA patients with liver segmental deformation were enrolled. The segmental deformation was assessed by conventional ultrasound followed with SWE examinations for evaluating the liver stiffness. Liver biopsy was performed in 11 patients in the region of SWE measurement and liver fibrosis was assessed using the Metavir classification. Aminotransferase to platelet ratio index (APRI) was calculated for comparison. The correlations between serum biochemical tests with SWE values were evaluated. Spearman’s rank coefficient test was performed to evaluate the correlation between variables. The SWE values of the biopsy segments had significant positive correlations with liver fibrosis severity (r = 0.828, p = 0.001), which was better than APRI (r = 0.366, p = 0.242). The levels of bilirubin and transaminase showed significant correlations with the SWE values at hypertrophic segments in all patients (r from 0.336 to 0.576, all p

To investigate the combined use of ultrasound together with clinical features to differentiate infantile hepatic hemangioma (IHH) from other focal liver lesions (FLLs) in children and to compare the efficacy of the combined method to that of CECT/MRI.The location, number, size and appearance of the tumors were evaluated in 45 children with IHH. Another 45 children with FLL were randomly selected as a control group. Independent factors for predicting IHH versus FLLs were evaluated. The diagnostic performance of the clinical and ultrasound features was compared with CECT/MRI.Compared with the control FLL group, the IHH group had a younger age at diagnosis (P = 0.008), lower alpha-fetoprotein (AFP) levels (P = 0.000), smaller lesion sizes (P = 0.000), and a higher tumor proportion with a resistance index (RI) of0.7. Multiple logistic regression analysis showed that age, size, RI and AFP were independent factors for predicting IHH. Receiver operating characteristic (ROC) curve analysis showed that the AUC (area under the curve) of the four combined independent factors was 0.881 (95% CI: 0.744-0.960), while the AUC for the CECT/MRI method was 0.905 (95% CI: 0.774-0.973), and the combined AUC for the independent factors and CECT/MRI was 0.929 (95% CI: 0.805-0.985). There were not statistically significant among the three AUCs (P 0.05).CECT/MRI was the effective diagnostic indicator for IHH. However, the combined clinical and ultrasound diagnoses, including age at diagnosis, lesion size, RI and AFP, can achieve the same effectiveness as CECT/MRI.

Aim To evaluate the technical feasibility and treatment results of ultrasound (US)-guided percutaneous radiofrequency ablation (RFA) combined with ethanol ablation (EA) for hepatocellular carcinoma (HCC) in the caudate lobe. Materials and methods The institutional review board approved this retrospective study and the requirement for informed consent was waived. From October 2009 to June 2016, there were 49 patients with HCC in the caudate lobe underwent ablation. Among them, 14 patients (12 men and two women, mean age: 49.9±14.9 years) with 16 caudate lobe HCCs (mean size: 3±1.1 cm) who underwent US-guided RFA-EA were identified. These patients were compared with patients who underwent RFA alone (n=17) and EA alone (n=18). Results HCCs in the RFA-EA group were larger than those in the EA-alone group (3±1.1 versus 2.1±0.7 cm, p=0.028). In the RFA-EA group, all the patients achieved treatment success (14/14) and entered follow-up. After a follow-up period of 17.1±10.6 months (range, 5–36 months), local tumour progression (LTP) was detected in four tumours. The 1- and 2-year LTP rates after RFA-EA were 18.8% and 27.8%, respectively. The 1- and 2-year disease-free survival (DFS) rates were 35.7% and 13.4%. Six patients died, with the 1-, 2-, and 3-year overall survival (OS) rates of 85.1%, 63.1%, and 21%, respectively. There were no significant differences in the LTP, DFS, and OS rates between the RFA-EA group and RFA-alone or EA groups (p=0.363–0.733). Conclusion US-guided percutaneous RFA-EA appears to be a feasible and effective treatment for HCC in the caudate lobe.

We aimed to investigate the feasibility of quantitative Sonazoid contrast-enhanced ultrasound (CEUS) in the diagnosis of biliary atresia (BA) at an early disease stage. First, liver tissue from 10 patients with BA and 10 patients with infantile hepatitis syndrome (IHS) was immunostained with cluster of differentiation 68 (CD68) to compare Kupffer cell counts. Then, quantitative Sonazoid CEUS was performed to evaluate the hyperplasia of the artery branches during the artery phase and changes in Kupffer cell counts during the Kupffer phase in a rat model of BA (common bile duct ligation). Based on our results, liver enhancement from Kupffer cells in BA patients was significantly higher than that in IHS patients (p = 0.0260.05). Furthermore, quantitative CEUS showed that the liver enhancement intensity of rats with bile duct ligation was substantially greater than that of rats without bile duct ligation during the artery phase 3 d post-operation and the Kupffer phase 5 d post-operation. The increased enhancement intensity in the Kupffer phase was in agreement with the increase in Kupffer cell counts in pathologic staining (18/20, 90%). In conclusion, quantitative Sonazoid CEUS can detect the hyperplasia of artery branches in artery phase and the increase of Kupffer cells in Kupffer phase in the liver of BA rats at an early disease stage. Therefore, Sonazoid CEUS could potentially be a valuable non-invasive alternative for the diagnosis of BA.

The aim of this study was to evaluate the influence of fasting time on the ultrasound identification and exclusion of biliary atresia in jaundiced infants through the use of the gallbladder classification scheme and to test the value of the gallbladder classification scheme in the diagnosis of biliary atresia in inexperienced individuals.A total of 188 jaundiced infants were enrolled in this study. All patients received detailed abdominal sonograms. Diagnoses were confirmed via liver biopsy, surgical findings, or follow-up. Infants were grouped into either the fasting group (fasting time ≥ 4 h) or the nonfasting group (fasting time 4 h). According to the gallbladder classification scheme, the gallbladders were classified into 4 types. The accuracy of diagnosing biliary atresia based on the gallbladder classification scheme was compared between the 2 groups. The gallbladders of 177 infants that were detectable by ultrasonography were used to compare the diagnostic performances between a junior radiologist and an experienced radiologist.The accuracies in the diagnosis of biliary atresia with the gallbladder classification scheme were 86.3% (82 of 95 patients) for the fasting group and 93.5% (87 of 93 patients) for the nonfasting group (P = .100). The diagnostic accuracies of biliary atresia were 88.7% (157 of 177 patients) for the junior radiologist and 90.4% (160 of 177 patients) for the experienced radiologist (P = .250). The κ value for the agreement between the 2 radiologists was 0.859.The performance of gallbladder diagnoses was not influenced by the fasting time with the use of the gallbladder classification scheme. Additionally, the gallbladder classification scheme may help junior radiologists to more effectively identify biliary atresia and nonbiliary atresia.

Focal liver lesions (FLLs) include a variety of benign and malignant tumors, and are usually differential diagnosed by ultrasonography. However, traditional two-dimensional contrast-enhanced ultrasound (2D-CEUS) can evaluate a single slice of a lesion. It cannot fully display perfusion of a lesion and the relationship with the surrounding tissues, which may lead to misdiagnosis. This study aimed to investigate real-time three-dimensional CEUS (3D-CEUS) for evaluation of the vascular configuration of FLLs. Using real-time 3D-CEUS with SonoVue on 161 patients with FLLs, we found that when compared with 2D-CEUS, real-time 3D-CEUS provided more detailed information about the origin, continuity, and numbers and structures of the feeding artery. On 3D-CEUS hepatocellular carcinoma showed intratumoral vessels with diffuse enhancement, metastatic liver cancer showed peritumoral vessels or absence of tumor vessels with ring-like enhancement or non-enhancement, focal nodular hyperplasia showed a spoke-wheel artery with diffuse enhancement, and hemangiomas showed nodular enhancement. Our results demonstrate that real-time 3D-CEUS using SonoVue is an effective method for visualizing the characteristic vascular configuration of FLLs, and this may assist with the diagnosis.

The objective of this study was to compare the clinical characteristics and imaging features on contrast-enhanced ultrasound (CEUS) of hepatitis B virus (HBV)-related combined hepatocellular–cholangiocarcinoma (CHC) and hepatocellular carcinoma (HCC). Thirty-one pathologically proven CHCs were included and 31 HCCs were randomly selected as controls. Elevated carbohydrate antigen (CA) 19-9 alone and simultaneous elevation of α-fetoprotein and CA19-9 were more frequent in CHC than in HCC patients (p = 0.004 and 0.029, respectively). On CEUS, homogeneous, heterogeneous and peripheral irregular rim-like enhancement was illustrated in 8 (25.8%), 12 (38.7%) and 11 (35.5%) CHCs and in 6 (19.4%), 23 (74.1%) and 2 (6.5%) HCCs, respectively (p = 0.007). Multivariate logistic regression analysis revealed CA19-9 elevation (p = 0.011, odds ratio [OR] = 6.545) and peripheral irregular rim-like enhancement on CEUS (p = 0.017, OR = 7.718) were independent variables. A receiver operating characteristic curve was plotted and the area under the curve was 0.740. CHC should be watched for in HBV-infected patients with liver tumor manifesting peripheral irregular rim-like enhancement on CEUS, accompanied by CA19-9 elevation.

Human nerve growth factor (hNGF) gene was proliferated with human leucocyte DNA as template by PCR. Then a fusion gene coding hNGF and luciferase (Luc) cDNAs was inserted into transfer vector pSXIVVI+X3/3 with the control of Syn XIV promoter. Luc and hNGF were simultaneously synthesized in Spodoptera larvae upon infection with a recombinant baculovirus, TnNPV-Luc-NGF-OCC+. Densitometric scanning of SDS-PAGE revealed that ∼18% of the total Coomassie blue-stainable protein of the infected larvae was represented by Luc protein, while the hNGF level was ∼8%. Both proteins were similar to their authentic counterparts in terms of immunoreactivity.

Using pSXIVVI + X3 as an expressing vector, an occluded recombinant Trichoplusia ni nuclear polyhedrosis virus carrying the cDNA encoding plasminogen activators inhibitor-2 (PAI-2) under the control of the Syn and XIV promoters, has been constructed. SDS-PAGE and immunoblot analysis revealed that the virus-mediated PAI-2, with a molecular weight of ∼45 kDa, was synthesized in the Sf cells at a level of ∼16% of total intracellular protein and in the supernatant phase at a level of ∼64% of total extracellular protein secreted into the hemolymph of infected larvae. The expressed protein was similar to its authentic counterpart in terms of immunoreactivity and bioactivity.

A fusion gene coding human granulocyte-macrophage colony stimulating factor (GM-CSF) and leukemia inhibitory factor (LIF) cDNAs was inserted into the transfer vector pSXIVVI+ X3 with the control of Syn and XIV promoters. The Sf9 cells (Spodoptera frugiperda) were co-transfected with the recombinant plasmid and TnNPV DNA (Trichoplusia ni nuclear polyhedrosis virus DNA). The fusion protein recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and leukemia inhibitory factor (LIF) could be synthesized in cells infected with recombinant virus at a level of about 23% of their total cellular protein. Activity analysis of the fusion protein in infected cells revealed that it exhibited the dual activities of GM-CSF and LIF. Western blot analysis of the expressed fusion protein in infected larvae showed that the virus-mediated fusion protein, with a molecular weight of approximately 35 kDa, is confirmed with immunoreactivity.