Objective:To explore the antitumor effect of ADU-S100/doxorubicin in situ vaccine on diffuse large B-cell lymphoma (DLBCL) and its mechanism.Methods:The 6-week-old female BALB/c mice were selected, and the bilateral murine subcutaneous B-cell lymphoma model was established with murine B-cell lymphoma A20 cells. The subcutaneous tumor-bearing mice were randomly divided into untreated group (without treatment), ADU-S100 in situ vaccine treatment group (intratumoral injection of interferon gene stimulating factor agonist ADU-S100), doxorubicin in situ vaccine treatment group (intratumoral injection of doxorubicin), and ADU-S100/doxorubicin in situ vaccine treatment group (intratumoral injection of ADU-S100 and doxorubicin) by using random number table method, with 5 mice in each group. The right tumors of the bilateral subcutaneous tumor-bearing mice were defined as proximal tumors, and the left tumors of the bilateral subcutaneous tumor-bearing mice were defined as distal tumors. Only the proximal tumors were treated via the intratumoral route, and the distal tumors were not treated. On day 23 after tumor inoculation, the percentages of CD11c + dendritic cells (DC), CD8 + CD11c + DC and CD80 + CD11c + DC in the spleen of mice in each group were detected by flow cytometry. The splenocytes of mice in each group were stimulated with A20 tumor cell lysate in vitro, the percentages of 5'-ethynyl-2'-deoxyuridine-positive (EdU +) cells and tumor necrosis factor-α-positive (TNF-α +) cells in CD8 + T cells in each in situ vaccine treatment group were detected by flow cytometry, and the killing effect of cytotoxic T lymphocyte (CTL) in each group was measured by using the lactate dehydrogenase (LDH) cytotoxicity assay kit. The mice treated with ADU-S100/doxorubicin in situ vaccine were intraperitoneally injected with anti-mouse CD8α (clone 53-6.7) mAb or isotype control on days 7, 12 and 17 after tumor inoculation to eliminate CD8 + cells. On day 23 after tumor inoculation, the proximal and distal tumor volumes of mice in the ADU-S100/doxorubicin in situ vaccine combined with anti-mouse CD8α (clone 53-6.7) mAb or isotype control treatment group were measured, the percentages of CD8 + T cells and CD8 + CD11c + DC in the spleen of tumor-bearing mice in these two groups were detected by flow cytometry, and the infiltration of CD8 + T cells in the tumor tissues from these two groups was detected by immunohistochemistry (IHC) staining. Results:On days 11, 14, 17, 20 and 23 after tumor inoculation, the proximal and distal tumor volumes of mice in each treated group were lower than those in the untreated group (all P < 0.05). The proportions of CD11c + DC in the spleen of the untreated group, ADU-S100 in situ vaccine treatment group, doxorubicin in situ vaccine treatment group and ADU-S100/doxorubicin in situ vaccine treatment group were (4.92±0.63)%, (7.54±0.84)%, (7.45±0.86)% and (11.63±0.85)%, respectively, and the difference was statistically significant ( F = 72.30, P < 0.001); the proportions of CD8 + CD11c + DC were (1.36±0.34)%, (4.02±0.43)%, (4.22±0.61)% and (6.11±0.73)%, respectively, and the difference was statistically significant ( F = 76.09, P < 0.001); the proportions of CD80 + CD11c + DC were (0.51±0.24)%, (1.69±0.23)%, (1.82±0.25)% and (4.09±0.39)%, respectively, and the difference was statistically significant ( F = 167.40, P < 0.001). The CTL responses and the proportion of EdU + cells and TNF-α + cells in CD8 + T cells in each in situ vaccine treatment group were higher than those in the untreated group (all P < 0.05). Furthermore, the enhanced CTL responses and the increased proportion of EdU + cells and TNF-α + cells in CD8 + T cells were observed in the ADU-S100/doxorubicin in situ vaccine treatment group as compared to the ADU-S100 in situ vaccine treatment group and doxorubicin in situ vaccine treatment group (all P < 0.05). The proportions of CD8 + T cells and CD8 + CD11c + DC in the spleen of mice treated with ADU-S100/doxorubicin in situ vaccine and anti-mouse CD8α mAb were lower than those in ADU-S100/doxorubicin in situ vaccine and isotype control group (both P < 0.05) and both proximal and distal tumor volumes of mice treated with ADU-S100/doxorubicin in situ vaccine and anti-mouse CD8α mAb were larger than those in ADU-S100/doxorubicin in situ vaccine and isotype control group (both P < 0.05). Conclusions:ADU-S100/doxorubicin in situ vaccine can induce profound regression of proximal tumors in bilateral murine subcutaneous B-cell lymphoma model and generate systemic immune responses capable of partially inhibiting distant tumor growth, and the antitumor efficacy of ADU-S100/doxorubicin in situ vaccine may require CD8 + CD11c + DC-mediated CD8 + T cell immune responses.

Objective To investigate the expression of zinc finger protein 382(ZNF382)in diffuse large B-cell lymphoma(DLBCL)tissue and its relationship with clinical pathological characteristics and prognosis of DLBCL patients.Methods A total of 57 DLBCL patients admitted to the Department of Hematology,Luoyang Central Hospital from January 2014 to December 2018 were selected as the research subjects.The biopsy pathological specimens and clinical data of DLBCL patients were collected;another 20 patients of reactive proliferative lymph node tissue preserved in the Department of Pathology,Luoyang Central Hospital were taken as the control group.The expression of ZNF382 in DLBCL tissue and reactive proliferative lymph node tissue was detected by En vision two-step method.The difference of ZNF382 expression was compared between DLBCL tissue and reactive proliferative lymph node tissue.The correlations of ZNF382 expression with the clinical features such as age,gender,primary tumor site,Ann Arbor stage,international prognostic index(IPI)score,Hans typing,B-symptoms,bone marrow infiltration,giant masses,Eastern Cooperative Oncology Group(ECOG)score,β2-microglobulin(β2-MG),serum lactate dehydrogenase(LDH),Ki67,and chemotherapy regimen of DLBCL patients were analyzed by univariate analysis;the survival curve was drawed by Kaplan Meier method,and the univariate and multivariate survival analysis were performed by log-rank tests and Cox proportional risk regression models.Results The expression level of ZNF382 in DLBCL tissue was significantly lower than that in reactive proliferative lymph node tissue(Z=-5.056,P＜0.01).The expression level of ZNF382 was correlated with IPI score,Ann Arbor stage,Hans typing,B-symptoms,bone marrow infiltration and giant masses of DLBCL patients(P＜0.05);the expression level of ZNF382 was not associated to gender,age,primary site,ECOG score,β2-MG,serum LDH,Ki67,and whether the chemotherapy regimen combined with rituximab or not of DLBCL patients(P＞0.05).Among the 57 DLBCL patients,the treatment was effective in 36 patients(63.20％)and ineffective in 21 patients(36.80％);the expression level of ZNF382 in tumor tissue of DLBCL patients with effective treatment was significantly higher than that of DLBCL patients with ineffective treatment(Z=-2.895,P＜0.05).The 2-year event free survival rate of DLBCL patients in the ZNF382 high expression group was significantly higher than that in the ZNF382 low expression group(x2=17.955,P＜0.001).The results of univariate survival analysis showed that female,primary lymph nodes,B-symptoms,bone marrow infiltration,giant masses,IPI score≥3,elevated β2-MG,Ki67＞70％,non-germinal center B-cell-like lymphoma,Ann Arbor stageⅢ-Ⅳ and low expression of ZNF382 were risk factors for poor prognosis in DLBCL patients(P＜0.05).The results of multivariate analysis showed that primary lymph nodes,Ann Arbor stage Ⅲ-Ⅳ and low expression of ZNF382 were independent influencing factors for poor prognosis in DLBCL patients(P＜0.05).Conclusion ZNF382 protein is low expressed in the tumor tissues of DLBCL patients,which is closely related to the occurrence,development and prognosis of DLBCL;and it can be used as an indicator for evaluating the prognosis of DLBCL.

Objective To explore whether stimulator of interferon genes(STING)agonist ADU-S100 could enhance the antitumor immune response of a chitosan(CS)nanoparticle-mediated DNA vaccine containing a tumor-specific antigen Dickkopf-related protein 1(DKK1)in multiple myeloma(MM).Methods CS-DNA nanoparticles were prepared by using the compound coprecipitation method.The particle sizes and Zeta potential of the CS-DNA nanoparticles were measured by using the Zetasizer Nano-ZS laser particle size analyzer.The DNA protection effect and in vivo DNA expression efficiency of the CS-DNA nanoparticles were assessed by using gel retardation assay and Western blot,respectively.The lentiviruses expressing human DKK1(hDKK1)genes were used to establish MPC-11 cells(MPC-11-hDKK1)which stably expressed hDKK1,and the MPC-11-hDKK1 cells were subcutaneously given to mice to construct tumor models.The tumor-bearing mice were randomly divided into a control group(intramuscular injection of CS-pcDNA3.1),an ADU-S100 immunization group(subcutaneous injection of ADU-S100),a CS-pDKK1 immunization group(intramuscular injection of CS-pDKK1)and an ADU-S1OO/CS-pDKK1 co-immunization group(intramuscular injection of CS-pDKK1+subcutaneous injection of ADU-S100),with 5 mice in each group.The tumor-bearing mice in each group were immunized 3 times at 10-day intervals according to the corresponding immunization schedule.The size of tumor was measured every week.On day 42 after MPC-11-hDKK1 cell inoculation,the tumor weight of mice in each immunization group was measured;the percentages of CD11c+dendritic cell(DC),CD8+CD11c+DC and major histocompatibility complex class Ⅱ(MHCII)+CD11c+DC subsets in the spleen of mice in each immunization group were detected by using flow cytometry.The splenocytes of mice in each group were stimulated with recombinant hDKK-1 protein in vitro,the percentage of EdU+cells in CD8+T lymphocytes in each immunization group was detected by using flow cytometry,and the killing effect of cytotoxic T lymphocyte(CTL)in each group was assessed by using the lactate dehydrogenase(LDH)cytotoxicity assay kit.Results The particle size and Zeta potential of the CS-DNA nanoparticles were(204.3±2.31)nm and(15.47±1.01)mV,respectively.Gel retardation assay showed that DNA enveloped in CS nanoparticles could be completely retarded.Western blot analysis indicated that CS-DNA nanoparticles could be effectively expressed in vivo.The relative expression of DKK1 protein was significantly higher in MPC-11-hDKK1 cells than in MPC-11-Ctrl cells(P＜0.05).On days 7 and 14 after MPC-11-hDKK1 cell inoculation,there was no significant difference in tumor volume of mice between the ADU-S100 immunization group,CS-pDKK1 immunization group,ADU-S100/CS-pDKK1 co-immunization group and the control group(P＞0.05);on days 21,28,35 and 42 after MPC-11-hDKK1 cell inoculation,the tumor volumes of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S100/CS-pDKK1 co-immunization group were significantly lower than those in the control group(P＜0.05);the tumor volume of mice in the ADU-S100/CS-pDKK1 co-immunization group was significantly lower than that in the ADU-S100 immunization group and CS-pDKK1 immunization group(P＜0.05).On day 42 after MPC-11-hDKK1 cell inoculation,the tumor weight of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S1 OO/CS-pDKK1 co-immunization group was significantly lower than that in the control group(P＜0.05);the tumor weight of mice in the ADU-S100/CS-pDKK1 co-immunization group was significantly lower than that in the ADU-S100 immunization group and CS-pDKK1 immunization group(P＜0.05).The proportions of CD11c+DC,CD8+CD11c+DC and MHCII+CD11c+DC subsets in the spleen of mice in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the control group(P＜0.05).The proportions of CD11c+DC,CD8+CD11c+DC and MHCII+CD11c+DC subsets in the spleen of mice in the ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the ADU-S100 immunization group and CS-pDKK1 immunization group(P＜0.05).The CTL killing effect and the proportion of EdU+cells in CD8+T lymphocytes in the ADU-S100 immunization group,CS-pDKK1 immunization group and ADU-S1OO/CS-pDKK1 co-immunization group were significantly higher than those in the control group(P＜0.05);the CTL killing effect and the proportion of EdU+cells in CD8+T lymphocytes in the ADU-S100/CS-pDKK1 co-immunization group were significantly higher than those in the ADU-S100 immunization group and CS-pDKK1 immunization group(P＜0.05).Conclusion STING agonist ADU-S100 can significantly improve the antitumor immunity of the CS-pDKK1 nanoparticle vaccine in MM,and this vaccine strategy provides a potential treatment approach for MM.

In recent years, with the changes in living standards and dietary structure, the incidence of non-alcoholic fatty liver disease has been increasing year by year in China, and the incidence rate in the general population is as high as 29.81%. An increasingly epidemiological evidence suggests that non-alcoholic fatty liver disease has become one of the causes of increasing liver cirrhosis and liver cancer. However, its etiology and pathogenesis are complex and have not yet been fully elucidated. Therefore, establishing an appropriate non-alcoholic fatty liver disease animal models for pre-clinical research is essential to elucidate its pathogenesis. This article summarizes the latest research progress of non-alcoholic fatty liver disease animal models, which are common at home and abroad in recent years.

Objective:To analyze the clinical characteristics of nephrogenic adenoma of the bladder.Methods:The clinical and pathological data of 8 patients with bladder nephrogenic adenoma, during the period from July 2016 to June 2019, were retrospectively analyzed. Patients’ age were 33 to 71 years old and the average age was 55, including 5 males and 3 females. The clinical manifestations were hematuria in 7 cases, urinary tract irritation in 6 cases, and no obvious symptoms in 1 case. There were 7 cases with cystitis, 3 cases with urolithiasis, 2 cases with bladder cancer, 1 case with invasive colonic mesentery fibroma, and 1 case without other complications. 5 cases had the history of urological operation. The predilection site was the lateral wall with 5cases; 5 cases were solitary; the average maximum diameter of the tumor was 0.9 cm (range 0.1-1.8 cm). Under cystoscope, papillary mass can be seen, the surface is bright red, the pedicle is not obvious, the papilla is thick and short, easy to bleed when touching; some of them are scattered and lichen like changes. All patients received transurethral resection of bladder mucosa.Results:Pathological examination shows that the bladder mucosa showed chronic inflammation, interstitial edema, granulation tissue hyperplasia, eosinophil infiltration and metaplasia of mesonephroid epithelium. All of the 8 patients were followed up by telephone for 2 to 38 months, with an average of 17.1 months. So far, neither recurrence has been detected.Conclusions:The diagnosis of nephrogenic adenoma of the bladder depends on pathological examination. It must be totally removed during operation. The recurrence and malignancy should be treated in time after operation.

Objective To observe MSCT features of partially or completely low enhancement of pancreas neuroendocrine tumors (PNENs).Methods The clinical data and MSCT features before pancreatectomy in 44 patients with confirmed PNENs were retrospectively reviewed.The MSCT findings were evaluated including tumor size, location, margin, density, intratumoral structure, bile duct and pancreatic ductal dilation and distant metastasis.Tumors were classified into complete enhancement type, partially or completely low enhancement type for further comparative analysis based on MSCT enhancement during pancreatic stage.Results A total of 56 PNENs in 44 patients were found, and there were 31 partially or completely low enhanced PNENs and 25 completely enhanced PNENs.The former were larger than the latter [mean tumor size, (3.3±2.2)cm vs (1.4±0.9) cm], and irregular shape and cystic components within tumors were more often observed (all P<0.05).There were no significant differences between the two types of PNENs in terms of gender, the presence of functional tumor, tumor location, clear tumor margin, intratumoral calcification, bile and pancreatic duct dilation and metastasis.76.0%(19/25) of completely enhanced PNENs reached peak enhancement in arterial phase, and 71.0%(22/31) of low enhancement PNENs reached peak in pancreatic phase.Enhanced intratumoral blood vessels in the arterial phase were more frequent in low enhancement PNENs, and the difference was statistically significant (P<0.05).There were significant differences on pathological grade between the two types of PNENs (G1=21,G2=4,G3=0 vs G1=18,G2=5, G3=8), and the difference was statistically significant (P<0.05).Conclusions Compared with complete enhancement PNENs, partially or completely low enhancement PNENs had bigger size, irregular shape, and cystic component.Intratumoral blood vessels in the arterial phase were observed, peak enhancement arrived later and the pathological grade was higher.

Objective To evaluate 3D CT features of normal anatomy,anatomic variations and fractures of occipital squama.Methods The 3D CT features on MIP,VR images were analyzed retrospectively in 589 pediatric cases.The normal anatomy,anatomic variations and fractures of occipital squama were observed respectively,and the differential diagnostic features including the individual location,appearance and extension were analyzed.Results Four hundred and thirty-three patients (75.2％) showed normal anatomy,including 154 patients with adult occipital anatomical features,279 patients with posterior intraoccipital synchondrosis,and 37 patients with Kerckring-supraoccipital synchondrosis.When cases with recent trauma history were excluded,113 patients (19.1％) showed anatomic variants,including unpenetrating sutures and penetrating sutures.The former could be subdivided to Mendosal sutures in 23 cases,superior median fissures in 19 cases,and midline supraoccipital fissures in 4 cases,while the latter could be subdivided to the interparietal bone variations in 54 cases,wormian bones in 23 cases,and accessory bones in 7 cases.Two or more variations coexisted in 33 cases.The occipital squama fractures were shown in 34 cases (5.6％),including linear fractures in 27 cases,comminuted fractures in 3 cases,with depression fracture in one case,separation of cranial sutures in 3 cases,and other fractures associated with variants in 3 cases.The fractures were sharp,or jagged,without limitation of the occification.Conclusion There are different 3D CT features of normal anatomy,anatomic variations and fractures of occipital squama in children,which are important for making the accurate diagnosis.

OBJECTIVETo establish a brain functional network of the whole brain based on glucose metabolism, and to evaluate the cost and efficiency of the functional network.

METHODS18F-FDG PET of 148 healthy volunteers (30-59 years, n = 148) was performed in a resting state. Images were registered to atlas by using the statistical parametric mapping (SPM) software. The functional connectivity between 90 cortical and sub-cortical regions was estimated by correlation analysis.

RESULTSGlucose metabolism brain functional networks had global efficiency greater than the lattice but less than the random graph, and local efficiency greater than random but less than lattice. This characteristically small-world behavior of the brain network was most consistently seen for low-cost to medium-cost networks. The small world regime was Cost-[0.0512, 0.5406]. The cost efficiency of the networks typically had a maximum positive value when the cost was 0.23.

CONCLUSIONSGlucose metabolism brain functional networks have economical small-world properties. It is feasible to analyze the functional characteristics of human brain by study 18F-FDG PET images. This paper provides a new method for studies on brain function.

Adult ; Brain ; metabolism ; physiology ; Brain Mapping ; Female ; Fluorodeoxyglucose F18 ; Glucose ; metabolism ; Humans ; Male ; Middle Aged ; Models, Neurological ; Nerve Net ; Positron-Emission Tomography ; methods

OBJECTIVE:To promote effective development of services in the intravenous drugs allocation centre and to bring the pharmacists'function into full play in the hospital pharmacy.METHODS:Information concerning services in the intravenous drugs allocation centre of our hospital and experiences from which was introduced,some problems and difficulties in the pharmacy services were analyzed.RESULTS&CONCLUSION:The centralized allocation and management of intra-venous drugs have ensured the safety and effectiveness of the clinical intravenous drug use and which have become the essential part of the pharmaceutical care with a core of rational drug use.

The discovery of the nerve growth factor(NGF),a specific protein material,by Rita Levi-Montalcini and her colleagues(1954)led to the observation that ifNGF is added to the culture medium,sympathetic and sensory neurons grow well inculture,for it is a potent and specific maintenance factor for these two types of neu-rons.Therefore,we chose the superior cervical ganglion(SCG)as a model systemfor nerve tissue culture in order to study the development,differentiation and rege-neration of nervous tissue in vitro.This report presents what we have observed onthe cultures of SCG that have been successfully established in our laboratory bymeans of explant technique.SCG from newborn rats were explanted onto collagen or plasma-coated coverslipsand maintained in Maximow depression slide assemblies at 37℃.The fluid me-dium was replaced twice a week and consisted of 1/3 calf serum,1/3 synthetic medium199 and 1/3 Hanks' BSS,or of equal parts of calf serum and Hanks' BSS,supplementedwith 600 mg% glucose.NGF(a crude extract of mouse salivary gland)was added tothe medium in a proportion of 1:20;for control,no NGF was added to the medium in some cultures.The cultures were examined microscopically every day,Nissl's stai-ned,Bodian's protargol and ammonium silver impregnated preparations were madeperiodically.Time lapse microcinematographic records were also made.We have planted altogether 209 SCG from newborn rats.All of them grew suc-cessfully in culture with NGF or without NGF,but the SCG with NGF grew muchmore rapidly than those without NGF.NGF strongly stimulates the outgrowth ofneurites and maintains the survival of the sympathetic neurons.Neurites grew outindividually or,more frequently,as bundles extending radially toward the peripheryor forming a meshwork.The tips of elongating neurites expanded to form growthcones from which are projected long slender microspikes(filopodia).These microspi-kes continually waved about,extending,and retracting as the growth cone movedover a substratum.The behavior of the growing tip of neurite is best analyzed bytime lapse microcinematography.The Schwann cells emerged from the explant andproliferate to accompany the neurites.These cells,fusiform or filiform in shape,were usually arranged in alignment along the neurites.They could be easily distingui-shed from fihroblasts.Mitosis of Schwann cells had been observed and recorded bytime lapse microcinematography.The bodies of the sympathetic neurons did not mig-rate away from the explant.