The degQ gene, amplified from Bacillus subtilis by PCR, was cloned to pUBS (sucrose induced secretion vector). After transformed into DB403, recombination named DB403(pUBSD) was formed. The results of the fermentation showed that degQ gene enhanced the expression of B. subtilis fibrin enzyme. The activity of the enzyme was increased to 2.2 times as the original one. In this article, the effects of different conditions, such as different kinds of sugar, different concentration of sucrose and different induced time were also be investigated and compared.

OBJECTIVETo assess the osseointegration capability of hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 (BMP-2) and hyaluronic acid to repair defects in the distal femur metaphysis in rabbits.

METHODSPorous titanium implants were made by sintering titanium powder at high temperature, which were coated with hydroxyapatite by alkali and heat treatment and with BMP-2 combined with bone regeneration materials. And hyaluronic acid was further used as delivery system to prolong the effect of BMP-2. The implants were inserted into the metaphysis of the distal femur of rabbits. The animals were killed at 6, 12 and 24 weeks to accomplish histological and biomechanical analyses.

RESULTSAccording to the result of histological analysis, the osseointegration in BMP-2 group was better than that of the HA-coated porous titanium group. In push-out test, all the samples had bigger shear stress as time passed by. There was statistical difference between the two groups in 6 and 12 weeks but not in 24 weeks.

CONCLUSIONHydroxyapatite-coated porous titanium with BMP-2 and hyaluronic acid has a good effect in repairing defects of distal femur in rabbits, which is a fine biotechnology for future clinical application.

Animals ; Biomechanical Phenomena ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Coated Materials, Biocompatible ; Durapatite ; Femur ; surgery ; Hyaluronic Acid ; pharmacology ; Osseointegration ; physiology ; Porosity ; Prostheses and Implants ; Rabbits ; Titanium ; Transforming Growth Factor beta ; pharmacology

Objective: To explore a single center large cohort of patients with Takayasu’s arteritis for their clinical manifestation and long-term outcome in China. Methods: We retrospectively analyzed 566 patients with Takayasu’s arteritis admitted in our hospital from 2002-01 to 2013-11 for their clinical characteristics, laboratory ifndings, angiographic features, treatment and long-term outcomes. Results: The patient’s ratio for female to male gender was 1 to 3.8 and the average onset age was (28.9 ± 12.0) years. The most common non-speciifc symptom, initial symptom and complication were fever (52/566 patients, 9.2%), dizziness (214 patients, 37.8%) and hypertension (392 patients, 69.3%) respectively. The patients with pulmonary artery and coronary artery involvement were 83 (14.7%) and 66 (11.7%) respectively, and 131 (23.1%) patients had faster erythrocyte sedimentation rate. The major vascular damage was steno-occlusive lesion and the most common involvement was left sub-clavian artery, which was observed in 278 (49.1%) patients. The treatments were mainly included in medication, interventional therapy, autologous blood vessel transplantation, artiifcial blood vessel transplantation and aortic valve replacement. There were 32 patients died during the mean follow-up period of (5.0 ± 0.2) years. Hypertension, complication and the progressive stage of disease were the major factors affecting prognosis in relevant patients (regression coefifcients: 4.664, 1.959 and 1.870 respectively, allP<0.05). Conclusion: Hypertension was the leading reason for patients’ hospital visit. Takayasu’s arteritis was closely related to cardiovascular disease, the early diagnosis and treatment were really important in clinical practice.

Objective To explore a kind of simple and high efficient approach to differentiate human embryonic stem cells(hESCs)into neural stem cells(NSCs).Methods hESCs were cultured in bacterial culture dish filled with serum free medium to gain embryoid bodies.Then the mature embryoid bodies grew in the special medium including B27 and noggin by adherent culture to differentiate into NSCs. Results The hESCs kept floating in the bacterial culture dish and growing all the time and then formed mature embryoid bodies 7 to 10 days later.The embryoid bodies could be differentiated into highly pure (96.4%)nestin positive cells.And these cells were differentiated into all kinds of neural cells if cultured further.Conclusions This kind of method is less time-consuming,cheaper,and more efficient than those of the results in literatures reported.It affords very good source of seed cells for cell transplantation therapy in the future.

Objective:To verify the predictive value of fully-automated 3D volume segmentation of CT images for the overall survival prognosis of resectable pancreatic ductal adenocarcinoma (PDAC).Methods:From July 2018 to March 2019, the clinical data of 198 cases of resectable PDAC were continuously collected in the First Affiliated Hospital of Naval Medical University. According to the level of carbohydrate antigen(CA)19-9 and carcinoembryonic antigen(CEA), the patient were divided into low CA19-9 group(≤210 U/ml ), high CA19-9 group (>210 U/ml ), normal CEA group (<5 ng/ml ) and high CEA group (≥5 ng/ml). Using our fully-automated segmentation tool developed in the early stage, images at the plain phase and portal phase were matched to those at the late artery phase by taking the artery phase as the matching target to establish UNet model; and the PDAC tumor and pancreatic glands were three-dimensionally segmented to estimate the tumor 3D volume. Univariate and multivariate logistic regression analysis were performed to compare the tumor 3D volume with the common preoperative risk factors (tumor 2D long diameter, CA19-9 level, CEA level, etc.) in predicting the patients′ survival. C-index was used to estimate the accuracy for predicting the survival. Receiver operating characteristics curve (ROC) was drawn and AUC was calculated to evaluate the accuracy for predicting the 1-year and 2-year overall survival and the influence of CA19-9 and CEA level on the patients′ overall survival.Results:Univariate logistic analysis showed that age, tumor 3D volume, tumor location, CA19-9 and CEA level were correlated with the patients′ overall survival. Multivariate logistic analysis showed that tumor 3D volume, CA199 and CEA were correlated with the overall survival. Among them, tumor 3D volume was most strongly correlated with the overall survival ( HR=2.25, 95% CI1.49-3.39, P<0.0001). The prognostic C-index of automatic 3D tumor volume, tumor long diameter, serum CEA and CA19-9 was 0.667(95% CI0.617-0.717), 0.637(0.583-0.691), 0.593(0.527-0.659) and 0.585(0.526-0.644), respectively. The AUCs of 3D tumor volume, tumor location, tumor long diameter, serum CEA and CA19-9 for predicting 1-year and 2-year survival were 0.726 and 0.698, 0.562 and 0.562, 0.703 and 0.660, 0.583 and 0.624, 0.602 and 0.609 respectively. C-index and AUC of tumor 3D volume was significantly better than those of the other common preoperative risk factors, and the difference was statistically significant (all P value <0.05). The survival of patients with large tumor 3D volume was greatly poorer than that of patients with small tumor 3D volume in low CA19-9 group, high CA19-9 group, normal CEA group and high CEA group, and the differences were all statistically significant ( HR=2.27, 95% CI 1.39-3.72; HR=2.42, 95% CI1.23-4.74; HR=2.08, 95% CI1.07-4.06; HR=2.67, 95% CI1.63-4.38, all P value <0.01). And the automatic 3D volume was the strongest predictor for the survival in high CA19-9 group. Conclusions:The tumor 3D volume obtained by automatic CT segmentation was an objective and reliable prognostic biomarker, which can supplement the established preoperativel risk factors and was expected to guide the personalized choice of neoadjuvant therapy.

Objective:To investigate the imaging features of colloid carcinoma arising from intraductal papillary mucinous neoplasm (IPMN) of pancreas and the differentiation features from ductal adenocarcinoma arising from IPMN, using the pathological findings as the reference.Methods:Twenty-four patients with pathologically confirmed colloid carcinoma from November 2013 to January 2020 in Changhai Hospital, Navy Medical University were included in this study. The clinical manifestations, imaging features and pathological data were retrospective reviewed. Thirty patients of ductal adenocarcinoma arising from IPMN confirmed by pathology were selected as the control group. CT and MRI features of two groups were blindly analyzed by two radiologists, including the lesions location, type of IPMN, size, components, density or signal, calcification, dilation and size of the main pancreatic duct (MPD), pancreatic parenchymal atrophy, fistula formation. The χ 2 test or Fisher exact probability was used to compare the imaging features between the two groups. Results:As for IPMN with colloid carcinoma, 16 cases were located in the head of the pancreas, 7 cases in the body and tail of the pancreas, and 1 case showed diffused changes of the pancreas. Mass was found in twenty-two cases, with the size of 54.5 (29) mm. Nineteen cases were solid-cystic, 4 were cystic and 1 was solid. Thick wall and internal separation with mild enhancement were displayed. Five cases were found with high signal on T 1WI. Thirteen cases had calcification and 2 cases had gas in the tumor. The size of MPD was (13±5) mm. Pancreatic parenchymal atrophy was found in 21 cases and fistula formation was found in 8 cases. The mass size of IPMN with colloid carcinoma was significantly greater than that of IPMN with ductal adenocarcinoma [31 (16) mm, Z=-3.758, P<0.001]. Solid-cystic mass was more found in IPMN with colloid carcinoma and solid mass was more found in IPMN with ductal adenocarcinoma ( P<0.001). Calcification ( P=0.001), fistula formation ( P=0.031), and high signal on T 1WI ( P=0.034) were more found in IPMN with colloid carcinoma than IPMN with ductal adenocarcinoma. Conclusion:Compared with IPMN with ductal adenocarcinoma, the solid-cystic mass, calcification, fistula formation and high signal on T 1WI were more commonly found in IPMN with colloid carcinoma.

The particularity of pancreatic anatomical location, the complexity of secretory function, and the diversity of pathology lead to complex imaging findings of pancreatic tumors. The common pancreatic tumors include pancreatic ductal adenocarcinoma, solid pseudopaillary neo-plasm, neuroendocrine neoplasm, intraductal papillary mucinous neoplasm, serous cystadenoma and mucinous cystic neoplasm. Atypical imaging findings are important reasons for misdiagnosis. Based on relevant clinical experiences, the authors analyze and summarize the atypical imaging findings of six kinds of common pancreatic tumors, aiming to improve radiologists and clinicians comprehensive understanding of pancreatic tumors.

Objective: To evaluate the efficiency and safety of low intensity conditional regimen for children with Fanconi anemia (FA) receiving allogenic hematopoietic stem cells transplantation (allo-HSCT). Methods: Four patients diagnosed as Fanconi anemia were enrolled in this study. One patient received HLA-identical sibling donor hematopoietic stem cell transplantation, two patients underwent unrelated donor matched (UD) HSCT, and one patient received unrelated cord blood transplantation. The conditional regimen consisted of Busulfan with low dose of cyclophosphamide. Results: All 4 cases succeeded in allo-HSCT. The median time for neutrophils engraftment was 11(9-15) day, median time to platelets (PLT) engraftment was 12 (8-28) day. One case occurred with grade I of aGVHD, 1 case with hemorrhagic cystitis. No patient happened with hepatic veno-occlusive disease (VOD). Conclusion: Low intensity of conditional regimen is efficient and safe which should be recommended for FA patients with HSCT.
Busulfan ; Fanconi Anemia ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation Conditioning

This study was performed to investigate the impact of nobiletin(NOB)on fracture healing in osteoporosis(OP)rats through the stimulator of interferon gene(STING)/nuclear transcription factor kappa B(NF-κB)signal pathway.A rat model of OP fracture was established by ovariectomy and right femoral shaft fracture intramedullary fixation;the rats after modeling were randomly grouped into model group,high dose(NOB-H,30 mg/kg NOB),medium dose(NOB-M,20 mg/kg NOB),low dose(NOB-L,10 mg/kg NOB)NOB group and NOB-H+ STING activator(DMXAA)group(30 mg/kg NOB+25 mg/kg DMXAA),and 18 rats experienced only ovaries expose were used as sham operation group.After the intervention,the fracture healing status of rats were measured;Micro-CT was used to detect the changes of bone trabecular microstructure in rats;commercial kits were used to detect the serum levels of bone metabolism related indicators(alkaline phosphatase(ALP),calcium,phosphorus)and inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β));HE was used to detect the morphological changes and trabecular area of femur,while Western blot was applied to detect the expression of STING/NF-κB pathway related proteins.Compared with the control group,the fracture line in the model group was clear,the trabecular structure was disordered and the gap was large,furthermore,the levels of TNF-α and IL-1β,the expression of STING and p-NF-κB p65/NF-κB p65 were significantly increased,the trabecular area,the levels of ALP,calcium,phosphorus,and bone mineral density(BMD),bone volume fraction(BV/TV),bone trabecular number(Tb.N)and bone trabecular thickness(Tb.Th)were significantly decreased(P<0.05);compared with the model group,the fracture line of NOB-L group,NOB-M group and NOB-H group gradually blurred,the trabecular structure arranged orderly,and the gap gradually decreased,and the trend of the index changes mentioned above were opposite to that of the model group(P<0.05).STING activators attenuated the promotion of fracture healing by NOB in OP rats and increased the inflammatory responses.In conclusion,NOB can reduce inflammatory reaction and promote fracture healing in OP rats,which may be related to the inhibition of STING/NF-κB signal pathway.

Objective:To investigate the clinical and imaging features of pancreatic intraductal oncocytic papillary neoplasm (IOPN).Methods:From January 2011 to August 2021, at the First Affiliated Hospital (Changhai Hospital) of Naval Medical University, 12 patients pathologically diagnosed with pancreatic IOPN after surgical resection were enrolled. Before operation, all patients underwent plain and enhanced computed tomography (CT) or magnetic resonance imaging (MRI). The clinical data (general conditions, main complaints, tumor related indicators and past medical history), CT and MRI features, surgical methods and pathologic results of the 12 patients with pancreatic IOPN were retrospectively analyzed. Descriptive method was used for statistical analysis.Results:Among 12 pancreatic IOPN patients, there were 7 males and 5 females, aged (54.0±13.0) years old (ranged from 31 to 75 years old). The symptoms were abdominal pain in 3 cases, jaundice in 1 case and 8 cases were detected during regular health checkups. Serum carbohydrate antigen 19-9 increased in 3 cases and carcinoembryonic antigen increased in 2 cases. One pancreatic IOPN patient with pancreatitis history and 3 pancreatic IOPN patients with diabetes history. Six cases were with the lesions located in the head of pancreas, 5 cases were located in the body and tail of pancreas and 1 case were diffused in the all the pancreas. Five cases were branch duct type, 2 cases were main duct type and 5 cases were mixed duct type. Ten pancreatic IOPN patients presented cystic or cystic-solid tumor, the maximum diameter (range) of the tumor was (50.3±31.1) mm (28 to 127 mm). The cyst walls of 6 patients were thickened and those of 9 patients were found with enhanced mural nodule or solid component, and none of them were growing outside the cystic wall. Two patients presented solid tumor located in the dilated pancreatic duct, and the maximum diameter (range) of the tumor was (25.5±0.5) mm (25 to 26 mm). The solid tumor demonstrated as slightly lower density on plain CT scan, lower signal on T1-weighted MRI imaging, high signal on T2-weighted MRI imaging, and limited diffusion on diffusion weighted imaging, and mild enhancement after CT and MRI enhanced scan. The main pancreatic duct dilated in 11 cases, and the inner diameter (range) was (10.5±8.1) mm (3 to 28 mm). The pancreatic parenchymal of 4 pancreatic IOPN patients was atrophy, 4 patients with calcification and 1 patient with lymphadenopathy. None of the 12 pancreatic IPON patients had peripheral blood vessel and tissue invasion. Six cases were received pancreaticoduodenectomy, 4 cases were underwent distal pancreatectomy, 2 cases underwent total pancreatectomy. The pathological classification of 7 pancreatic IOPN patients was invasive carcinoma, 4 cases were with high-grade dysplasia and 1 case with low-grade dysplasia.Conclusion:The clinical features of pancreatic IOPN are atypical and the imaging findings are mostly solid or cystic-solid tumor, pancreatic duct dilation, solid component of tumor located in the dilated pancreatic duct, and no peripheral tissue invasion.