Objective To observe and compare the atomic force microscopy (AFM) images of Yersinia pestis EV76 and the changes in the morphology of the bacteria treated with normal serum and F1 antibody from rabbit,and to explore the immunoassay method to detect Yersinia pestis by AFM. Methods The Yersinia pestis were treated with normal serum and F1 antibody from rabbit and control buffer. All the prepared samples were observed and analyzed by AFM. The changes in the cell surface structures were probed and characterized through sectional analysis,especially the changes of Ra and Rq value. Results The normal morphology of Yersinia pestis was oval in shape with a relatively smooth surface, the size dimension of which was about 1.1-1.3 μm in length with a section profile of 0.8-1.0 μm in width and 0.04-0.06 μm in step height. The step height of the bacteria treated with the normal serum and F1 antibody was obviously enlarged. The shape of the bacteria treated with F1 antibody changed irregularly. Furthermore, the surface of the bacteria was more roughened. Conclusion The morphological characters of Yersinia pestis has been acquired through its AFM images. The morphology of Yersinia pestis treated with F1 antibody has changed greatly, and the index of roughness can be regarded as the distinguished index to detect Yersinia pestis by AFM.

Objective To investigate the activation, distribution, origin, and expression of hepatic stem cells(HSC)in different histological types of primary liver carcinomas. Methods The histological and immunohistochemical features of 94 cases of hepatocellular carcinoma (HCC), 12 cases of intrahepatic cholangiocarcinoma (ICC) and 10 cases of mixed hepatocarcinoma were examined by HE staining and immunohistochemistry SP method, with 5 cases of sclerotic liver and 4 cases of normal liver tissues as control. Results HSC expression was observed and the transfor mation from HSC to carcinoma cell was also noted in the liver. CK7, CK19, c-kit, Thy-1, and AFP were found expressed in different types of hepatic carcinomas and the greatest intensive expression was found in the mixed hepatocarcinoma (P

ObjectiveTo synthesize 99Tcm labeled hydrazine-nicotinamide ( HYNIC)-c (RGDfK)and evaluate its biodistribution and imaging in the severe combined immunodeficiency (SCID) nude mice bearing human lung adenocarcinoma.Methods( 1 )Tcm-HYNIC-c(RGDfK) was prepared by a two-step method using tricine and ethylenediamine diacetate (EDDA) as coligands and HYNIC as the dual functional chelator.The bioactivity of 99Tc m-HYNIC-c (RGDfK) was measured by cell binding experiments.(2) The nude mice bearing human A549 lung adenocarcinoma were randomly divided into 7 groups with 5 in each group.The 7 th group was the competitive inhibition control group and was administrated 100 μg HYNIC -c (RDGfK) 30 min earlier before the injection of 99Tcm-H Y N IC-c ( RGDfK ).The nude mice were scanned at 0.5,1,2,4,8 and 12 h respectively after intravenous injection of 7.4 MBq 99Tcm-HYNIC-c(RGDfK).The biodistribution of the agent was measured as ％ ID/g.The uptake ratio of tumor to muscle (T/NT) was also measured by placing ROI on 99Tcm-HYNIC-c(RGDfK) SPECT imaging.(3)Gamma imaging was performed in 6 mice including 3 in the competitive inhibition control group at 0.5,1,2,4,8 and 12 h post injection.ResultsThe labeling yield of 99Tcm-HYNIC-c(RGDfK) was more than 90％,and the radiochemical purity was more than 95％.99Tcm-HYNIC-c(RGDfK) can specifically bind with A549 adenocarcinoma cells with a binding rate up to 36.14％.Biodistribution study showed that the uptake in the kidney was above 20 ％ ID/g during 0.5 - 8 h post injection.The ％ ID/g in tumor was 10.52 ± 1.48 at 0.5 h,17.26 ±2.81 at 8 h,and 8.93 ±0.90 at 12 h.However,the ％ ID/g in tumor was only 2.29 ±0.85 in the competitive inhibition control group at 0.5 h.The highest T/NT was 6.87 at 8 h by the ROI analysis.Xenograffted tumors could be visualized at 1 h and delineated more clearly from 4 to 8 h post injection of 99Tcm-HYNIC-c(RGDfK).Conclusions99 Tcm-HYNIC-c (RGDfK) can be readily synthesized.Its binding with A549 lung adenocarcinoma cells is specific and the binding rate is high.

Objective:To explore the clinical characteristics and prognostic factors of children with relapsed acute lymphoblastic leukemia (ALL).Methods:The clinical data of 52 children with relapsed ALL in Children's Hospital of Shanxi Province from January 2010 to April 2019 were retrospectively analyzed. The clinical characteristics of the children were summarized and the prognostic factors after recurrence were analyzed.Results:Till May 1, 2019, 5 out of 52 children gave up treatment after diagnosis and were lost to follow-up. For the remaining 47 children with successful follow-up, the median age at initial diagnosis was 60 months (11-168 months), the median time from initial diagnosis to relapse was 21 months (2-112 months), the median follow-up time was 5.5 months (1.0-69.0 months), and the 2-year overall survival (OS) rate after relapse was 31%. Nine patients accepted allogeneic hematopoietic stem cell transplantation after the second time complete remission, the median time from diagnosis to transplantation was 4.5 months (3.0-7.0 months), and the median follow-up time was 22 months (4-69 months). The 2-year OS rates in relapsed children with white blood cell count < 50×10 9/L and ≥ 50×10 9/L at initial diagnosis were 39% and 13%, respectively (χ 2=5.623, P=0.018). The 2-year OS rate after relapse in standard-risk, intermediate-risk and high-risk groups were 72%, 31% and 8%, respectively (χ 2=10.068, P=0.007). The 2-year OS rate after relapse in very early relapse, early relapse and late relapse groups were 0, 33% and 79%, respectively (χ 2=30.066, P < 0.01). The 2-year OS rate after relapse in chemotherapy with or without radiotherapy group, transplantation group and irregular treatment group were 57%, 89% and 0, respectively (χ 2=26.885, P < 0.01). Cox multivariate analysis showed that relapse time was the independent risk factor affecting the prognosis of children with relapsed ALL ( HR=0.340, 95% CI 0.146-0.789, P=0.012). Compared with the transplantation group, the risk of death in the chemotherapy with or without radiotherapy group and the irregular treatment group was significantly higher ( HR=12.313, 95% CI 1.266-119.758, P=0.031; HR=20.699, 95% CI 2.230-192.129, P=0.008), suggesting that hematopoietic stem cell transplantation is a protective factor for the prognosis of children with relapsed ALL. Conclusions:The relapse of ALL in children mainly happens in very early and early time. The main part of relapse is bone marrow, and there are many high-risk patients at initial diagnosis. The risk group at initial diagnosis, white blood cell count at initial diagnosis, relapse time, and treatment after relapse are the risk factors affecting the prognosis, and the relapse time and hematopoietic stem cell transplantation are the independent prognostic factors.

BACKGROUND: Recently, some people believed that the mechanisms of primary hepatic carcinoma might be caused by poor differentiation or disdifferentiation of hepatic stem cells. Studies on hepatic stem cells are in the early stage at present, and the theory of "stem cell origins" of human primary hepatic carcinoma deserves further verification. OBJECTIVE: To investigate the activation, distribution, origin and immunological expression characteristics of hepatic stem cells in different histopathologic types of primary hepatic carcinoma. DESIGN: Observational comparative study. SETTING: Tumor Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA. PARTICIPANTS: Experiments were performed at the Laboratory of Tumor Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from September 2003 to July 2004. We took 94 cases of hepatic cellular cancer, 12 cases of intrahepatic cholangiocellular carcinoma and 10 cases of mixed hepatocarcinoma paraffin-embedded tissue blocks as research objects, with 5 cases of liver cirrhosis and 4 cases of normal liver as experimental control. These materials were collected from the archive of the Department of Pathology of Daping Hospital. Primary hepatic carcinoma tissues and corresponding adjacent liver tissues were obtained from patients who had undergone surgery for the removal of their tumors. All the patients were not treated by chemotherapy or radiotherapy before the operation. They had signed the informed consent. Main Antibodies were bought from Santa Cruz Company.METHODS: The histological and immunohistochemical characteristics were examined by haematoxylin and eosin staining and immunohistochemistry (SP method), including mouse antihuman cytokeratin 19 monoclonal antibody, mouse antihuman cytokeratin 7 monoclonal antibody, mouse antihuman cytokeratin 8&&18 monoclonal antibody, mouse antihuman c-kit monoclonal antibody, mouse antihuman Thy-1 monoclonal antibody, mouse antihuman alpha fetoprotein monoclonal antibody. MAIN OUTCOME MEASURES: Expression of immunological markers of hepatic stem cells in different histopathologic types. RESULTS: Immunological markers of hepatic stem cells expressed variously in different histopathologic types of primary hepatic carcinoma. Hepatic stem cells differentiated into hepatoma carcinoma cells in all the types. The highest expression rate of hepatic stem cell immunophenotype was found in the mixed hepatocarcinoma (P < 0.05). Immunophenotypes of hepatic stem cells were negative in normal group and cirrhosis group. CONCLUSION: Hepatic stem cells of varied differentiations and origins existed in different histopathologic types of primary hepatic carcinoma.

Objective To construct the adeno-associated virus (AAV)vector of mouse carrying T-bet gene,which was applied to mouse model of asthma by nose,and to investigate the immunomodulatory effects of T-bet delivery on asthma.Methods Forty healthy Balb/c mice (aged 6-8 weeks) were randomly divided into 4 groups:control group (group A),asthma model group(group B),model/control recombinant adeno-associated virus carrying enhanced green fluorescent protein(rAAV-eGFP) intervention group (group C) and model/rAAV-T-bet virus intervention group(group D),with 10 mice in each group.In group B,group C,and group D,Balb/c mice were sensitized by intraperitoneal injection with OVA and challenged by nebulized OVA.In group C and group D,Balb/c mice were intervened with the isodose rAAV-eGFP and rAAV-T-bet,while in group B,the mice were intervened with the same amount of saline,both of them were dropped into the nasal cavity.Twenty-four hours after the last injection,the mice were sacrificed and the samples were obtained.The total cells in bronchoalveolar lavage fluid (BALF) were counted,and the types of cells were analyzed by Wright-Giemsa staining.The levels of interferon-γ (IFN-γ),IL-4 and IL-5 in BALF were determined by enzyme-linked immunosorbent assay.The expressions of T-bet,GATA-3 and Foxp3 protein in the lungs of asthmatic mice were detected with immunohistochemical technique.Results 1.The level of specific transcription factor T-bet in group B and group C were distinctly lower than that of group A (all P ＜ 0.05),while the level of GATA-3 was significantly higher than that of group A(all P ＜ 0.05).In group D,the expression of T-bet protein was stronger than that in group B and group C,however,the expression of GATA-3 decreased obviously (all P ＜ 0.05).The results of Foxp3 were similar to T-bet.2.In group D,the mice had suppressed levels of IL-4 [(158 ± 55) ng/L] and IL-5 [(68-± 22) ng/L] compared with those in group B and group C(all P ＜0.05).The level of IFN-γ[(113-±35) ng/L] increased compared with those in group B and group C (all P ＜ 0.05).At the same time,the number of total cells and eosinophil in BALF were both obviously lower than those of group B and group C (all P ＜ 0.05).Conclusions On the basis of building mouse model of asthma,intranasal administration of rAAV-T-bet can deliver T-bet gene to airway successfully and efficiently,and plays an immunomodulatory role in immune confusion of asthma.

Objective:To evaluate the short-term efficacy and the improvement of quality of life of enzyme replacement therapy (ERT) with Imiglucerase on children with Gaucher disease(GD) through the same time monitoring.Methods:Six children diagnosed as GD who were treated by ERT with Imiglucerase in the Department of Hematology of the Children′s Hospital of Shanxi Province from May 2019 to May 2020 were recruited.Every 3 months, the sizes of the liver and spleen was palpated, the change of bone pain was recorded, and the haematological index was examed.The volumes of the liver and spleen at 1-year treatment were measured by CT.Bone involvement was examined by magnetic resonance imaging (MRI). In addition, the body weight, height, and the 36-Item Short Form Survey (SF-36) were measured and compared with pre-treatment levels.These data were analyzed statistically by SPSS 25.0 and the difference between pretherapy and post-treatment was compared by paired t test. Results:Six children diagnosed as GD received ERT with Imiglucerase.No adverse events were reported.Decreased volumes of the liver and spleen, and increased hemoglobin level and platelet count were detected after 3-6 months of ERT.After 1 year of ERT, hemoglobin level significantly increased compared with pre-treatment level ( t=4.200, P=0.008). Although the platelet count increased at 1-year ERT, it was comparable with pre-treatment level ( t=2.260, P=0.073). The volumes of liver and spleen decreased by (22.10±15.28)% ( t=2.725, P=0.042) and (47.10±18.42)% ( t=3.162, P=0.034) after 1 year of ERT, respectively.During the first year of ERT, the height and weight increased (6.17±2.86) cm ( t=5.286, P=0.003) and (4.08±2.01) kg ( t=4.975, P=0.004), respectively.SF-36 score increased significantly from (489.35±103.99) points to (632.75±73.34) points ( t=5.740, P=0.002). After 1 year of ERT, 1 patient still had bone pain, and 2 cases were worse in bone MRI, which may be attributed to the short period of follow-up and insufficient dose, and another 3 had no change in bone MRI. Conclusions:ERT ameliorates GD-associated anemia, organomegaly and growth retardation, and improves the growth rate of body mass and height and the quality of life in the short period.However, short-term ERT does not improve the bone disease.

OBJECTIVETo confirm the amount of fertilizer and the ration of fertilizer.

METHODThrough the experiment of planting Salvia miltiorrhiza in pot and in field were carried out.

RESULTWhen N:P = 1:1, The production was 1.8 times more than the contrast group in plotting experiment, and in field the production of the highest fertilizer plot were 2.5 times more than the contrast and the higher and high fertilizer plot is 2.25 and 1.2 times respective were than those in the contrast group.

CONCLUSIONThe proper ration of nitrogenous fertilizer and phosphorous fertilizer is 1:1. Nitrogen shows negative effects to the accumulation of tanshinon IIA, the more the nitrogen ous fertilizer, The less the content of tanshinon IIA. In contrast, phosphor ous fertilizer shows good effects on the accumulation of tanshinon IIA. Phosphorous fertilizer could alleviate the decline of the content of tanshinon IIA by using nitrogenous fertilizer. The accumulation peak of the tanshinon IIA emerge in the period of 150 d.

Diterpenes, Abietane ; Fertilizers ; Nitrogen ; analysis ; pharmacology ; Phenanthrenes ; analysis ; Phosphorus ; analysis ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; growth & development ; Salvia miltiorrhiza ; chemistry ; growth & development

The aim of the study is through observing the morphology of the prepared influenza virus (H1N1) with transmission electron microscopy (TEM) and atomic force microscopy (AFM) to explore the application of AFM on the research of the external character of viruses and provide a new, simple and efficient technique for the study of the viral morphology. TEM image was obtained by negatively stained influenza virus with 1% Phosphotungstic Acid; AFM image applied the tapping mode to influenza virus without any further treatment in air at room temperature, and the morphology parameters, including length (diameter), Ra and Rq are calculated by sectional analysis. The shapes of influenza virus A are spherical, filamentous or other pleomorphous particles observed by both AFM and TEM. TEM image of influenza virus A is two-dimensional image, and viral surface has visible spikes, while AFM exhibits the three-dimensional image that can be described with several quantifiable indexes through sectional analysis. AFM phase images show viral surface clearly which is characterized by rugged feature and gear-like protuberance. As compared with TEM, AFM is a new research tool for viral morphology study with the advantages of simple sample preparing, visible interface and is intuitionistic for researchers. The surface characteristic parameters of viruses provided by AFM can be served as the main quantifiable indexes for viral morphological study.
Influenza A Virus, H1N1 Subtype ; ultrastructure ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission

OBJECTIVES: We analyzed the chromosomal-arm-level copy number alterations (CNAs) in the cervical exfoliative cell and tissue samples by using the low-coverage whole genomic sequencing technique. METHODS: In this study, we retrospectively collected 55 archived exfoliated cervical cell suspension samples and the corresponding formalin-fixed and paraffin-embedded tissue section samples including 27 invasive cervical cancer and 28 control cases. We also collected 19 samples of the cervical exfoliative cells randomly from women to verify the new algorithm model. We analyzed the CNAs in cervical exfoliated cell and tissue samples by using the low-coverage next generation of sequencing. RESULTS: In the model-building study, multiple chromosomal-arm-level CNAs were detected in both cervical exfoliated cell and tissue samples of all cervical cancer cases. By analyzing the consistency of CNAs between exfoliated cells and cervical tissue samples, as well as the heterogeneity in individual patient, we also established a C-score algorithm model according to the chromosomal-arm-level changes of 1q, 2q, 3p, 7q. The C-score model was then validated by the pathological diagnosis of all 74 exfoliated cell samples (including 55 cases in model-building group and 19 cases in verification group). In our result, a cutoff value of C-score > 6 showed 100% sensitivity and 100% specificity in the diagnosis of cervical cancer. CONCLUSION: In this study, we found that CNAs of cervical exfoliated cell samples could robustly distinguish invasive cervical cancer from cancer-free tissues. And we have also developed a C-score algorithm model to process the sequencing data in a more standardized and automated way.
Diagnosis ; DNA Copy Number Variations ; Female ; Genome* ; High-Throughput Nucleotide Sequencing ; Humans ; Mass Screening ; Population Characteristics ; Retrospective Studies ; Sensitivity and Specificity ; Uterine Cervical Neoplasms*